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Sample records for human bone hydroxyapatite

  1. Quantification of bone ingrowth into porous block hydroxyapatite in humans.

    PubMed

    Ayers, R A; Wolford, L M; Bateman, T A; Ferguson, V L; Simske, S J

    1999-10-01

    This study sought to quantify bone ingrowth from a single bone-implant surface into porous block hydroxyapatite used in maxillofacial applications. Seventeen maxillary hydroxyapatite implants (implant time of 4-138 months, 39-month mean) were harvested for analysis from 14 patients. The implants had been placed into the lateral maxillary wall during orthognathic surgery, juxtapositioned to the maxillary sinus. Ingrowth was measured in 100-microm increments from a bone-implant interface to a depth of 1500 microm. Bone ingrowth averaged over the 14 patients (0-1100 microm depth) is described by the equation % ingrowth - 20% * (depth in millimeters) + 41.25% (R2 = 0.98, n = 10 incremental depths). Beyond 1100 microm, the average ingrowth remained constant at 15.0 +/- 0.7%. The duration of implantation also showed as affect on the percent ingrowth into the implants at the incremental depths, and the percent ingrowth asymptotically approached a maximum. Overall, the composite average data from all depths is best described by the logarithmic function % ingrowth = 15% * ln(implantation time in months) - 24.0% (R2 = 0.71, n = 14 patients). Several factors may come into play in determining bone ingrowth including the mechanical environment, the osteoconductivity of the implant material, and the osteogenic capability of the tissues in the pore spaces. Measurements of bone ingrowth are most influenced by the depth into the implant and the time the implant was in the body; the age of the patient had little affect on bone ingrowth. PMID:10400880

  2. Lamellar Spacing in Cuboid Hydroxyapatite Scaffolds Regulates Bone Formation by Human Bone Marrow Stromal Cells

    PubMed Central

    Afghani, Shahrzad; Franco, Jaime; Launey, Max; Marshall, Sally; Marshall, Grayson W.; Nissenson, Robert; Lee, Janice; Tomsia, Antoni P.; Saiz, Eduardo

    2011-01-01

    Background A major goal in bone engineering is the creation of large volume constructs (scaffolds and stem cells) that bear load. The scaffolds must satisfy two competing requirements—they need be sufficiently porous to allow nutrient flow to maintain cell viability, yet sufficiently dense to bear load. We studied the effect of scaffold macroporosity on bone formation and scaffold strength, for bone formed by human bone marrow stromal cells. Methods Rigid cubical hydroxyapatite/tricalcium phosphate scaffolds were produced by robo-casting. The ceramic line thickness was held constant, but the distance between adjacent lines was either 50, 100, 200, 500, or 1000 μm. Cultured human bone marrow stromal cells were combined with the scaffolds in vitro; transplants were placed into the subcutis of immunodeficient mice. Transplants were harvested 9, 18, 23, 38, or 50 weeks later. Bone formation and scaffold strength were analyzed using histology and compression testing. Results Sixty transplants were evaluated. Cortical bone increased with transplant age, and was greatest among 500 μm transplants. In contrast, maximum transplant strength was greatest among 200 μm transplants. Conclusions Lamellar spacing within scaffolds regulates the extent of bone formation; 500 μm yields the most new bone, whereas 200 μm yields the strongest transplants. PMID:21294634

  3. Cell-induced response by tetracyclines on human bone marrow colonized hydroxyapatite and Bonelike.

    PubMed

    Gomes, P S; Santos, J D; Fernandes, M H

    2008-05-01

    Semi-synthetic tetracyclines are commonly used antibiotics that also seem to play an important role in the modulation of the immuno-inflammatory imbalance, verified in several bone diseases. The association of a therapeutic agent (that prevents bacterial infection and induces tissue formation) to a biomaterial aiming to repair/regenerate bone defects could contribute to a more predictable clinical outcome. The present study intends to evaluate the proliferation and functional activity of osteoblast-induced human bone marrow cells, cultured on the surface of hydroxyapatite (HA) and Bonelike, in the presence of therapeutic concentrations of doxycycline and minocycline. First passage bone marrow cells were cultured for 35 days on the surface of HA and Bonelike discs, in the absence or presence of 1 microg ml(-1) doxycycline and minocycline. Cultures performed in standard tissue culture plates were used as control. Doxycycline or minocycline induced cell proliferation and increased the extent of matrix mineralization in osteoblastic cell cultures established in the three substrates. Also, an improved biological behavior was verified in seeded Bonelike compared with HA. The results suggest that the local delivery of tetracyclines might associate the antimicrobial activity in implant-related bone infection with an eventual induction of osteoblastic proliferation and maintenance of the characteristic biological activity of these cells. PMID:18291737

  4. First histological observations on the incorporation of a novel nanocrystalline hydroxyapatite paste OSTIM® in human cancellous bone

    PubMed Central

    Huber, Franz-Xaver; Belyaev, Orlin; Hillmeier, Joachim; Kock, Hans-Juergen; Huber, Colette; Meeder, Peter-Juergen; Berger, Irina

    2006-01-01

    Background: A commercially available nanocrystalline hydroxyapatite paste Ostim® has been reported in few recent studies to surpass other synthetic bone substitutes with respect to the observed clinical results. However, the integration of this implantable material has been histologically evaluated only in animal experimental models up to now. This study aimed to evaluate the tissue incorporation of Ostim® in human cancellous bone after reconstructive bone surgery for trauma. Methods: Biopsy specimens from 6 adult patients with a total of 7 tibial, calcaneal or distal radial fractures were obtained at the time of osteosynthesis removal. The median interval from initial operation to tissue sampling was 13 (range 3–15) months. Samples were stained with Masson-Goldner, von Kossa, and toluidine blue. Osteoid volume, trabecular width and bone volume, and cortical porosity were analyzed. Samples were immunolabeled with antibodies against CD68, CD56 and human prolyl 4-hydroxylase to detect macrophages, osteoblasts, and fibroblasts, respectively. TRAP stainings were used to identify osteoclasts. Results: Histomorphometric data indicated good regeneration with normal bone turnover: mean osteoid volume was 1.93% of the trabecular bone mass, trabecular bone volume – 28.4%, trabecular width – 225.12 μm, and porosity index – 2.6%. Cortical and spongious bone tissue were well structured. Neither inflammatory reaction, nor osteofibrosis or osteonecrosis were observed. The implanted material was widely absorbed. Conclusion: The studied nanocrystalline hydroxyapatite paste showed good tissue incorporation. It is highly biocompatible and appears to be a suitable bone substitute for juxtaarticular comminuted fractures in combination with a stable screw-plate osteosynthesis. PMID:16762071

  5. Polymer-ceramic spiral structured scaffolds for bone tissue engineering: effect of hydroxyapatite composition on human fetal osteoblasts.

    PubMed

    Zhang, Xiaojun; Chang, Wei; Lee, Paul; Wang, Yuhao; Yang, Min; Li, Jun; Kumbar, Sangamesh G; Yu, Xiaojun

    2014-01-01

    For successful bone tissue engineering, a scaffold needs to be osteoconductive, porous, and biodegradable, thus able to support attachment and proliferation of bone cells and guide bone formation. Recently, hydroxyapatites (HA), a major inorganic component of natural bone, and biodegrade polymers have drawn much attention as bone scaffolds. The present study was designed to investigate whether the bone regenerative properties of nano-HA/polycaprolactone (PCL) spiral scaffolds are augmented in an HA dose dependent manner, thereby establishing a suitable composition as a bone formation material. Nano-HA/PCL spiral scaffolds were prepared with different weight ratios of HA and PCL, while porosity was introduced by a modified salt leaching technique. Human fetal osteoblasts (hFOBs) were cultured on the nano-HA/PCL spiral scaffolds up to 14 days. Cellular responses in terms of cell adhesion, viability, proliferation, differentiation, and the expression of bone-related genes were investigated. These scaffolds supported hFOBs adhesion, viability and proliferation. Cell proliferation trend was quite similar on polymer-ceramic and neat polymer spiral scaffolds on days 1, 7, and 14. However, the significantly increased amount of alkaline phosphatase (ALP) activity and mineralized matrix synthesis was evident on the nano-HA/PCL spiral scaffolds. The HA composition in the scaffolds showed a significant effect on ALP and mineralization. Bone phenotypic markers such as bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC), and type I collagen (Col-1) were semi-quantitatively estimated by reverse transcriptase polymerase chain reaction analysis. All of these results suggested the osteoconductive characteristics of HA/PCL nanocomposite and cell maturation were HA dose dependent. For instance, HA∶PCL = 1∶4 group showed significantly higher ALP mineralization and elevated levels of BSP, ON, OC and Col-I expression as compared other lower or higher ceramic ratios

  6. Polymer-Ceramic Spiral Structured Scaffolds for Bone Tissue Engineering: Effect of Hydroxyapatite Composition on Human Fetal Osteoblasts

    PubMed Central

    Zhang, Xiaojun; Chang, Wei; Lee, Paul; Wang, Yuhao; Yang, Min; Li, Jun; Kumbar, Sangamesh G.; Yu, Xiaojun

    2014-01-01

    For successful bone tissue engineering, a scaffold needs to be osteoconductive, porous, and biodegradable, thus able to support attachment and proliferation of bone cells and guide bone formation. Recently, hydroxyapatites (HA), a major inorganic component of natural bone, and biodegrade polymers have drawn much attention as bone scaffolds. The present study was designed to investigate whether the bone regenerative properties of nano-HA/polycaprolactone (PCL) spiral scaffolds are augmented in an HA dose dependent manner, thereby establishing a suitable composition as a bone formation material. Nano-HA/PCL spiral scaffolds were prepared with different weight ratios of HA and PCL, while porosity was introduced by a modified salt leaching technique. Human fetal osteoblasts (hFOBs) were cultured on the nano-HA/PCL spiral scaffolds up to 14 days. Cellular responses in terms of cell adhesion, viability, proliferation, differentiation, and the expression of bone-related genes were investigated. These scaffolds supported hFOBs adhesion, viability and proliferation. Cell proliferation trend was quite similar on polymer-ceramic and neat polymer spiral scaffolds on days 1, 7, and 14. However, the significantly increased amount of alkaline phosphatase (ALP) activity and mineralized matrix synthesis was evident on the nano-HA/PCL spiral scaffolds. The HA composition in the scaffolds showed a significant effect on ALP and mineralization. Bone phenotypic markers such as bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC), and type I collagen (Col-1) were semi-quantitatively estimated by reverse transcriptase polymerase chain reaction analysis. All of these results suggested the osteoconductive characteristics of HA/PCL nanocomposite and cell maturation were HA dose dependent. For instance, HA∶PCL = 1∶4 group showed significantly higher ALP mineralization and elevated levels of BSP, ON, OC and Col-I expression as compared other lower or higher ceramic ratios

  7. Growth and osteogenic differentiation of alveolar human bone marrow-derived mesenchymal stem cells on chitosan/hydroxyapatite composite fabric.

    PubMed

    Kim, Beom-Su; Kim, Jin Seong; Chung, Yong Sik; Sin, Yeon-Woo; Ryu, Kyung-Ho; Lee, Jun; You, Hyung-Keun

    2013-06-01

    Scaffolds can be used for tissue engineering because they can serve as templates for cell adhesion and proliferation for tissue repair. In this study, chitosan/hydroxyapatite (CS/HAp) composites were prepared by coprecipitation synthesis. Then, CS and CS/HAp fabrics were prepared by wet spinning. CS fibers with a diameter of 15 ± 1.3 μm and CS/HAp fibers with a diameter of 22 ± 1.2 μm were successfully produced; incorporation of HAp into the CS/HAp fibers was confirmed by X-ray diffraction analysis. Biological in vitro evaluations showed that human mesenchymal stem cells (hMSCs) cultured on CS/HAp fabric showed increased proliferation compared to those cultured on pure CS fabric, which was observed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, DNA content assay, and [(3) H] thymidine incorporation assay. Neither the CS nor CS/HAp scaffold exhibited any cytotoxicity to hMSCs, as shown by viability staining and cytotoxicity fluorescence image assays. After 10 days of culturing, the attachment of cells onto the scaffold was observed by scanning electron microscopy. Furthermore, under osteogenic differentiation conditions, alkaline phosphatase (ALP) activity and calcium accumulation was higher in cells cultured on the CS/HAp scaffold than in cells cultured on the CS scaffold. The mRNA expression of osteoblast markers, including ALP, osteocalcin, Co1Ia1, and runt-related transcription factor 2, was higher in cells cultured on CS/HAp than in cells cultured on the CS fabric. The results of this study indicate that the CS/HAp composite fabric may serve as a good scaffold for bone tissue engineering applications. PMID:23135904

  8. The induction of bone formation by smart biphasic hydroxyapatite tricalcium phosphate biomimetic matrices in the non-human primate Papio ursinus

    PubMed Central

    Ripamonti, U; Richter, P W; Nilen, R W N; Renton, L

    2008-01-01

    Long-term studies in the non-human primate Chacma baboon Papio ursinus were set to investigate the induction of bone formation by biphasic hydroxyapatite/p-tricalcium phosphate (HA/β-TCP) biomimetic matrices. HA/β-TCP biomimetic matrices in a pre-sinter ratio (wt%) of 40/60 and 20/80, respectively, were sintered and implanted in the rectus abdominis and in calvarial defects of four adult baboons. The post-sinter phase content ratios were 19/81 and 4/96, respectively. Morphological analyses on day 90 and 365 showed significant induction of bone formation within concavities of the biomimetic matrices with substantial bone formation by induction and resorption/dissolution of the implanted matrices. One year after implantation in calvarial defects, 4/96 biphasic biomimetic constructs showed prominent induction of bone formation with significant dissolution of the implanted scaffolds. The implanted smart biomimetic matrices induce de novo bone formation even in the absence of exogenously applied osteogenic proteins of the transforming growth factor-β(TGF-β) superfamily. The induction of bone formation biomimetizes the remodelling cycle of the cortico-cancellous bone of primates whereby resorption lacunae, pits and concavities cut by osteoclastogenesis are regulators of bone formation by induction. The concavities assembled in HA/β-TCP biomimetic bioceramics are endowed with multifunctional pleiotropic self-assembly capacities initiating and promoting angiogenesis and bone formation by induction. Resident mesenchymal cells differentiate into osteoblastic cell lines expressing, secreting and embedding osteogenic soluble molecular signals of the TGF-β superfamily within the concavities of the biomimetic matrices initiating bone formation as a secondary response. PMID:18363843

  9. Human recombinant cementum attachment protein (hrPTPLa/CAP) promotes hydroxyapatite crystal formation in vitro and bone healing in vivo.

    PubMed

    Montoya, Gonzalo; Arenas, Jesús; Romo, Enrique; Zeichner-David, Margarita; Alvarez, Marco; Narayanan, A Sampath; Velázquez, Ulises; Mercado, Gabriela; Arzate, Higinio

    2014-12-01

    Cementum extracellular matrix is similar to other mineralized tissues; however, this unique tissue contains molecules only present in cementum. A cDNA of these molecules, cementum attachment protein (hrPTPLa/CAP) was cloned and expressed in a prokaryotic system. This molecule is an alternative splicing of protein tyrosine phosphatase-like A (PTPLa). In this study, we wanted to determine the structural and functional characteristics of this protein. Our results indicate that hrPTPLa/CAP contains a 43.2% α-helix, 8.9% β-sheet, 2% β-turn and 45.9% random coil secondary structure. Dynamic light scattering shows that this molecule has a size distribution of 4.8 nm and aggregates as an estimated mass of 137 kDa species. AFM characterization and FE-SEM studies indicate that this protein self-assembles into nanospheres with sizes ranging from 7.0 to 27 nm in diameter. Functional studies demonstrate that hrPTPLa/CAP promotes hydroxyapatite crystal nucleation: EDS analysis revealed that hrPTPLa/CAP-induced crystals had a 1.59 ± 0.06 Ca/P ratio. Further confirmation with MicroRaman spectrometry and TEM confirm the presence of hydroxyapatite. In vivo studies using critical-size defects in rat cranium showed that hrPTPLa/CAP promoted 73% ± 2.19% and 87% ± 1.97% new bone formation at 4 and 8 weeks respectively. Although originally identified in cementum, PTPLa/CAP is very effective at inducing bone repair and healing and therefore this novel molecule has a great potential to be used for mineralized tissue bioengineering and tissue regeneration. PMID:25263524

  10. Thermal Diffusivity in Bone and Hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Calderón, A.; Peña Rodríguez, G.; Muñoz Hernández, R. A.; Díaz Gongora, J. A. I.; Mejia Barradas, C. M.

    2004-09-01

    We report thermal diffusivity measurements in bull bone and commercial hydroxyapatite (HA), both in powder form, in order to determinate the thermal compatibility between these materials. Besides this, we report a comparison between these measured values and those of metallic samples frequently used in implants, as high purity titanium and stainless steel. Our results show a good thermal compatibility (74%) between HA and bone, both in powder form. Finally, it was obtained a one order of magnitude difference between the thermal diffusivity values of metallic samples and those corresponding values to bone and HA being this difference greater in titanium than in stainless steel, which is important to consider in some biomedical and dental applications.

  11. Incorporation of nanostructured hydroxyapatite and poly(N-isopropylacrylamide) in demineralized bone matrix enhances osteoblast and human mesenchymal stem cell activity.

    PubMed

    Nicoletti, Alessandro; Torricelli, Paola; Bigi, Adriana; Fornasari, Piermaria; Fini, Milena; Moroni, Lorenzo

    2015-01-01

    Demineralized bone matrix (DBM) is currently used in many clinical applications for bone augmentation and repair. DBM is normally characterized by the presence of bone morphogenetic proteins. In this study, the authors have optimized methods to obtain DBM under good manufacturing practice, resulting in enhanced bioactivity. The processed DBM can be used alone, together with nanostructured hydroxyapatite (nanoHA), or dispersed in a physiological carrier or hydrogel. In this study, osteoblasts (MG-63) and human bone marrow derived mesenchymal stem cells (hMSCs) were cultured on DBM pastes made in phosphate buffered saline solution or poly(N-isopropylacrylamide) (PNIPAAM) hydrogels with or without nanoHA. The authors observed that the presence of PNIPAAM reduced osteoblast adhesion, while the addition of nanoHA increased osteoblast adhesion, proliferation, interleukin-6 (IL-6) production, and reduced lactate dehydrogenase (LDH) production. Increasing concentrations of PNIPAAM in combination with nanoHA further increased osteoblast proliferation, and decreased IL-6 and LDH production. Incorporation of PNIPAAM in DBM enhanced hMSCs proliferation and collagen type-I production. Furthermore, a combination of PNIPAAM and nanoHA further increased alkaline phosphatase and osteocalcin production in hMSCs, independently from the concentration of PNIPAAM. This study shows that combinations of DBM with nanoHA and PNIPAAM seem to offer a promising route to enhance cell activity and induce osteogenic differentiation. PMID:26443012

  12. BMP2-loaded hollow hydroxyapatite microspheres exhibit enhanced osteoinduction and osteogenicity in large bone defects

    PubMed Central

    Xiong, Long; Zeng, Jianhua; Yao, Aihua; Tu, Qiquan; Li, Jingtang; Yan, Liang; Tang, Zhiming

    2015-01-01

    The regeneration of large bone defects is an osteoinductive, osteoconductive, and osteogenic process that often requires a bone graft for support. Limitations associated with naturally autogenic or allogenic bone grafts have demonstrated the need for synthetic substitutes. The present study investigates the feasibility of using novel hollow hydroxyapatite microspheres as an osteoconductive matrix and a carrier for controlled local delivery of bone morphogenetic protein 2 (BMP2), a potent osteogenic inducer of bone regeneration. Hollow hydroxyapatite microspheres (100±25 μm) with a core (60±18 μm) and a mesoporous shell (180±42 m2/g surface area) were prepared by a glass conversion technique and loaded with recombinant human BMP2 (1 μg/mg). There was a gentle burst release of BMP2 from microspheres into the surrounding phosphate-buffered saline in vitro within the initial 48 hours, and continued at a low rate for over 40 days. In comparison with hollow hydroxyapatite microspheres without BMP2 or soluble BMP2 without a carrier, BMP2-loaded hollow hydroxyapatite microspheres had a significantly enhanced capacity to reconstitute radial bone defects in rabbit, as shown by increased serum alkaline phosphatase; quick and complete new bone formation within 12 weeks; and great biomechanical flexural strength. These results indicate that BMP2-loaded hollow hydroxyapatite microspheres could be a potential new option for bone graft substitutes in bone regeneration. PMID:25609957

  13. Strontium and zoledronate hydroxyapatites graded composite coatings for bone prostheses.

    PubMed

    Boanini, Elisa; Torricelli, Paola; Sima, Felix; Axente, Emanuel; Fini, Milena; Mihailescu, Ion N; Bigi, Adriana

    2015-06-15

    Both strontium and zoledronate (ZOL) are known to be useful for the treatment of bone diseases associated to the loss of bone substance. In this work, we applied an innovative technique, Combinatorial Matrix-Assisted Pulsed Laser Evaporation (C-MAPLE), to deposit gradient thin films with variable compositions of Sr-substituted hydroxyapatite (SrHA) and ZOL modified hydroxyapatite (ZOLHA) on Titanium substrates. Compositional gradients were obtained by simultaneous laser vaporization of the two distinct material targets. The coatings display good crystallinity and granular morphology, which do not vary with composition. Osteoblast-like MG63 cells and human osteoclasts were co-cultured on the thin films up to 21 days. The results show that Sr counteracts the negative effect of relatively high concentration of ZOL on osteoblast viability, whereas both Sr and ZOL enhance extracellular matrix deposition. In particular, ZOL promotes type I collagen production, whereas Sr increases the production of alkaline phosphatase. Moreover, ZOL exerts a greater effect than Sr on osteoprotegerin/RANKL ratio and, as a consequence, on the reduction of osteoclast proliferation and activity. The deposition method allows to modulate the composition of the thin films and hence the promotion of bone growth and the inhibition of bone resorption. PMID:25706198

  14. Hydroxyapatite-binding peptides for bone growth and inhibition

    DOEpatents

    Bertozzi, Carolyn R.; Song, Jie; Lee, Seung-Wuk

    2011-09-20

    Hydroxyapatite (HA)-binding peptides are selected using combinatorial phage library display. Pseudo-repetitive consensus amino acid sequences possessing periodic hydroxyl side chains in every two or three amino acid sequences are obtained. These sequences resemble the (Gly-Pro-Hyp).sub.x repeat of human type I collagen, a major component of extracellular matrices of natural bone. A consistent presence of basic amino acid residues is also observed. The peptides are synthesized by the solid-phase synthetic method and then used for template-driven HA-mineralization. Microscopy reveal that the peptides template the growth of polycrystalline HA crystals .about.40 nm in size.

  15. Cytotoxic Effects and Osteogenic Activity of Calcium Sulfate with and without Recombinant Human Bone Morphogenetic Protein 2 and Nano-Hydroxyapatite Adjacent to MG-63 Cell Line

    PubMed Central

    Ghorbanzadeh, Abdollah; Aminsobhani, Mohsen; Khoshzaban, Ahad; Abbaszadeh, Armin; Ghorbanzadeh, Atiyeh; Shamshiri, Ahmad Reza

    2015-01-01

    Objectives: The aim of this study was to assess the cytotoxic effects and osteogenic activity of recombinant human bone morphogenetic protein (rhBMP2) and nano-hydroxyapatite (n-HA) adjacent to MG-63 cell line. Materials and Methods: To assess cytotoxicity, the 4,5-dimethyl thiazolyl-2,5-diphenyl tetrazolium bromide (MTT) assay was used. Alkaline phosphatase (ALP) activity and osteogenic activity were evaluated using Alizarin red and the von Kossa staining and analyzed by one-way ANOVA followed by Tukey’s post hoc test. Results: The n-HA/calcium sulfate (CS) mixture significantly promoted cell growth in comparison to pure CS. Moreover, addition of rhBMP2 to CS (P=0.02) and also mixing CS with n-HA led to further increase in extracellular calcium production and ALP activity (P=0.03). Conclusion: This in vitro study indicates that a scaffold material in combination with an osteoinductive material is effective for bone matrix formation. PMID:26877731

  16. Collagen-gelatin-genipin-hydroxyapatite composite scaffolds colonized by human primary osteoblasts are suitable for bone tissue engineering applications: in vitro evidences.

    PubMed

    Vozzi, G; Corallo, C; Carta, S; Fortina, M; Gattazzo, F; Galletti, M; Giordano, N

    2014-05-01

    The application of porous hydroxyapatite (HAp)-collagen as a bone tissue engineering scaffold represents a new trend of mimicking the specific bone extracellular matrix (ECM). The use of HAp in reconstructive surgery has shown that it is slowly invaded by host tissue. Therefore, implant compatibility may be augmented by seeding cells before implantation. Human primary osteoblasts were seeded onto innovative collagen-gelatin-genipin (GP)-HAp scaffolds containing respectively 10%, 20%, and 30% HAp. Cellular adhesion, proliferation, alkaline phosphatase (ALP) activity, osteopontin (OPN), and osteocalcin (OC) expressions were evaluated after 3, 7, 15, and 21 days. The three types of scaffolds showed increased cellular proliferation over time in culture (maximum at 21 days) but the highest was recorded in 10% HAp scaffolds. ALP activity was the highest in 10% HAp scaffolds in all the times of evaluation. OC and OPN resulted in higher concentration in 10% HAp scaffolds compared to 20% and 30% HAp (maximum at 21 days). Finally, scanning electron microscopy analysis showed progressive scaffolds adhesion and colonization from the surface to the inside from day 3 to day 21. In vitro attachment, proliferation, and colonization of human primary osteoblasts on collagen-GP-HAp scaffolds with different percentages of HAp (10%, 20%, and 30%) all increased over time in culture, but comparing different percentages of HAp, they seem to increase with decreasing of HAp component. Therefore, the mechanical properties (such as the stiffness due to the HAp%) coupled with a good biomimetic component (collagen) are the parameters to set up in composite scaffolds design for bone tissue engineering. PMID:23775901

  17. Preparation and characterization of hydroxyapatite/sodium alginate biocomposites for bone implant application

    NASA Astrophysics Data System (ADS)

    Kanasan, Nanthini; Adzila, Sharifah; Suid, Mohd Syafiq; Gurubaran, P.

    2016-07-01

    In biomedical fields, synthetic scaffolds are being improved by using the ceramics, polymers and composites materials to avoid the limitations of allograft. Ceramic-polymer composites are appearing to be the most successful bone graft substitute in human body. The natural bones itself are well-known as composite of collagen and hydroxyapatite. In this research, precipitation method was used to synthesis hydroxyapatite (HA)/sodium alginate (SA) in various parameters. This paper describes the hydroxyapatite/sodium alginate biocomposite which suitable for use in bone defects or regeneration of bone through the characterizations which include FTIR, FESEM, EDS and DTA. In FTIR, the characteristi peaks of PO4-3 and OH- groups which corresponding to hydroxyapatite are existed in the mixing powders. The needle-size particle of hydroxyapatite/ alginate (HA/SA) are observed in FESEM in the range of 15.8nm-38.2nm.EDS confirmed the existence of HA/SA composition in the mixing powders. There is an endothermic peak which corresponds to the dehydration and the loss of physically adsorbed water molecules of the hydroxyapatite (HA)/sodium alginate (SA) powder which are described in DTA.

  18. Ultrastructure of regenerated bone mineral surrounding hydroxyapatite-alginate composite and sintered hydroxyapatite.

    PubMed

    Rossi, Andre L; Barreto, Isabela C; Maciel, William Q; Rosa, Fabiana P; Rocha-Leão, Maria H; Werckmann, Jacques; Rossi, Alexandre M; Borojevic, Radovan; Farina, Marcos

    2012-01-01

    We report the ultrastructure of regenerated bone surrounding two types of biomaterials: hydroxyapatite-alginate composite and sintered hydroxyapatite. Critical defects in the calvaria of Wistar rats were filled with micrometer-sized spherical biomaterials and analyzed after 90 and 120 days of implantation by high-resolution transmission electron microscopy and Fourier transform infrared attenuated total reflectance microscopy, respectively. Infrared spectroscopy showed that hydroxyapatite of both biomaterials became more disordered after implantation in the rat calvaria, indicating that the biological environment induced modifications in biomaterials structure. We observed that the regenerated bone surrounding both biomaterials had a lamellar structure with type I collagen fibers alternating in adjacent lamella with angles of approximately 90°. In each lamella, plate-like apatite crystals were aligned in the c-axis direction, although a rotation around the c-axis could be present. Bone plate-like crystal dimensions were similar in regenerated bone around biomaterials and pre-existing bone in the rat calvaria. No epitaxial growth was observed around any of the biomaterials. A distinct mineralized layer was observed between new bone and hydroxyapatite-alginate biomaterial. This region presented a particular ultrastructure with crystallites smaller than those of the bulk of the biomaterial, and was possibly formed during the synthesis of alginate-containing composite or in the biological environment after implantation. Round nanoparticles were observed in regions of newly formed bone. The findings of this work contribute to a better understanding of the role of hydroxyapatite based biomaterials in bone regeneration processes at the nanoscale. PMID:22057083

  19. Gallium increases bone calcium and crystallite perfection of hydroxyapatite.

    PubMed

    Bockman, R S; Boskey, A L; Blumenthal, N C; Alcock, N W; Warrell, R P

    1986-12-01

    Gallium, a group IIIa metal, is known to interact with hydroxyapatite as well as the cellular components of bone. In recent studies we have found gallium to be a potent inhibitor of bone resorption that is clinically effective in controlling cancer-related hypercalcemia as well as the accelerated bone resorption associated with bone metastases. To begin to elucidate gallium's mechanism of action we have examined its effects on bone mineral properties. After short-term (14 days) administration to rats, gallium nitrate produced measurable changes in bone mineral properties. Using atomic absorption spectroscopy, low levels of gallium were noted to preferentially accumulate in regions of active bone formation, 0.54 +/- .07 microgram/mg bone in the metaphyses versus 0.21 +/- .03 microgram/mg bone in the diaphyses, P less than 0.001. The bones of treated animals had increased calcium content measured spectrophotometrically. Rats injected with radiolabeled calcium during gallium treatment had greater 45-calcium content compared to control animals. By wide-angle X-ray analyses, larger and/or more perfect hydroxyapatite was observed. The combined effects of gallium on bone cell function and bone mineral may explain its clinical efficacy in blocking accelerated bone resorption. PMID:3026592

  20. Osteocalcin enhances bone remodeling around hydroxyapatite/collagen composites.

    PubMed

    Rammelt, Stefan; Neumann, Mirjam; Hanisch, Uwe; Reinstorf, Antje; Pompe, Wolfgang; Zwipp, Hans; Biewener, Achim

    2005-06-01

    The effect of osteocalcin (OC), an extracellular bone matrix protein, on bone healing around hydroxyapatite/collagen composites was investigated. Cylindrical nanocrystalline hydroxyapatite implants of 2.5-mm diameter containing 2.5% biomimetically mineralized collagen type I were inserted press-fit into the tibial head of adult Wistar rats. To one implant group, 10 mug/g OC was added. Six specimens per group were analyzed at 2, 7, 14, 28, and 56 days. After 14 days, newly formed woven bone had reached the implant surface of the OC implants whereas a broad fibrous interface could still be observed around controls. Woven bone was formed directly around both implant groups after 28 days and had been replaced partially by lamellar bone around the OC implants only. No significant differences in total bone contact were seen between both groups after 56 days. The higher number of phagocytosing cells and osteoclasts characterized immunohistochemically with ED1, cathepsin D, and tartate-resistant alkaline phosphatase around the OC implants at the early stages of bone healing suggests an earlier onset of bone remodeling. The earlier and increased expression of bone-specific matrix proteins and multifunctional adhesion proteins (osteopontin, bone sialoprotein, CD44) at the interface around the OC implants indicates that OC may accelerate bone formation and regeneration. This study supports the observations from in vitro studies that OC activates both osteoclasts and osteoblasts during early bone formation. PMID:15800855

  1. Eggshell Derived Hydroxyapatite as Bone Graft Substitute in the Healing of Maxillary Cystic Bone Defects: A Preliminary Report

    PubMed Central

    Kattimani, Vivekanand S; Chakravarthi, P Srinivas; Kanumuru, Narasimha Reddy; Subbarao, Vummidisetti V; Sidharthan, A; Kumar, T S Sampath; Prasad, L Krishna

    2014-01-01

    Background: Since ancient times, use of graft materials to promote healing of defects of bone is wellknown. Traditionally, missing bone is replaced with material from either patient or donor. Multiple sources of bone grafts have been used to graft bone defects to stimulate bone healing. Hydroxyapatite is naturally occurring mineral component of bone, which is osteoconductive. This versatile biomaterial is derived from many sources. The aim of this study is to evaluate the efficacy of eggshell derived hydroxyapatite (EHA) in the bone regeneration of human maxillary cystic bone defects secondary to cystic removal/apicoectomy and compare the material properties of EHA in vitro. Materials and Methods: A total of eight maxillary bone defects were grafted after cystic enucleation and/or apicoectomy in the year 2008 and completed the study at 1 year. The patients were followed-up 2 weeks after surgery for signs and symptoms of infection or any other complications that may have been related to surgical procedure. Follow-up radiographs were obtained immediately after surgery followed by 1, 2, and 3 months to assess the efficacy of EHA in bone healing. Physicochemical characterization of the EHA was carried out in comparison with synthetic hydroxyapatite (SHA), also compared the biocompatibility of EHA using in vitro cytotoxicity test. Results: By the end of the 8th week, the defects grafted with EHA showed complete bone formation. However, bone formation in non-grafted sites was insignificant. The values of density measurements were equal or more than that of surrounding normal bone. These results indicate that the osseous regeneration of the bone defect filled with EHA is significant. EHA showed the superior material properties in comparison with SHA. Conclusion: EHA is a versatile novel bone graft substitute that yielded promising results. Because of its biocompatibility, lack of disease transfer risks, ease of use and unlimited availability, EHA remains a viable choice

  2. Bio-inspired mineralization of hydroxyapatite in 3D silk fibroin hydrogel for bone tissue engineering.

    PubMed

    Jin, Yashi; Kundu, Banani; Cai, Yurong; Kundu, Subhas C; Yao, Juming

    2015-10-01

    To fabricate hard tissue implants with bone-like structure using a biomimetic mineralization method is drawing much more attentions in bone tissue engineering. The present work focuses in designing 3D silk fibroin hydrogel to modulate the nucleation and growth of hydroxyapatite crystals via a simple ion diffusion method. The study indicates that Ca(2+) incorporation within the hydrogel provides the nucleation sites for hydroxyapatite crystals and subsequently regulates their oriented growth. The mineralization process is regulated in a Ca(2+) concentration- and minerlization time-dependent way. Further, the compressive strength of the mineralized hydrogels is directly proportional with the mineral content in hydrogel. The orchestrated organic/inorganic composite supports well the viability and proliferation of human osteoblast cells; improved cyto-compatibility with increased mineral content. Together, the present investigation reports a simple and biomimetic process to fabricate 3D bone-like biomaterial with desired efficacy to repair bone defects. PMID:26209967

  3. Cobalt doped proangiogenic hydroxyapatite for bone tissue engineering application.

    PubMed

    Kulanthaivel, Senthilguru; Roy, Bibhas; Agarwal, Tarun; Giri, Supratim; Pramanik, Krishna; Pal, Kunal; Ray, Sirsendu S; Maiti, Tapas K; Banerjee, Indranil

    2016-01-01

    The present study delineates the synthesis and characterization of cobalt doped proangiogenic-osteogenic hydroxyapatite. Hydroxyapatite samples, doped with varying concentrations of bivalent cobalt (Co(2+)) were prepared by the ammoniacal precipitation method and the extent of doping was measured by ICP-OES. The crystalline structure of the doped hydroxyapatite samples was confirmed by XRD and FTIR studies. Analysis pertaining to the effect of doped hydroxyapatite on cell cycle progression and proliferation of MG-63 cells revealed that the doping of cobalt supported the cell viability and proliferation up to a threshold limit. Furthermore, such level of doping also induced differentiation of the bone cells, which was evident from the higher expression of differentiation markers (Runx2 and Osterix) and better nodule formation (SEM study). Western blot analysis in conjugation with ELISA study confirmed that the doped HAp samples significantly increased the expression of HIF-1α and VEGF in MG-63 cells. The analysis described here confirms the proangiogenic-osteogenic properties of the cobalt doped hydroxyapatite and indicates its potential application in bone tissue engineering. PMID:26478356

  4. Conversion of bulk seashells to biocompatible hydroxyapatite for bone implants.

    PubMed

    Vecchio, Kenneth S; Zhang, Xing; Massie, Jennifer B; Wang, Mark; Kim, Choll W

    2007-11-01

    Strombus gigas (conch) shells and Tridacna gigas (Giant clam) shells have dense, tailored structures that impart excellent mechanical properties to these shells. In this investigation, conch and clam seashells were converted to hydroxyapatite (HAP) by a hydrothermal method at different temperatures and for different conversion durations. Dense HAP structures were created from these shells throughout the majority of the samples at the relative low temperature of approximately 200 degrees C. The average fracture stress was found to be approximately 137-218MPa for partially converted conch shell samples and approximately 70-150MPa for original and converted clamshell samples, which is close to the mechanical strength of compact human bone. This indicates that the converted shell samples can be used as implants in load-bearing cases. In vivo tests of converted shell samples were performed in rat femoral defects for 6 weeks. The microtomography images at 6 weeks show that the implants did not move, and untreated control defects remain empty with no evidence of a spontaneous fusion. Histological study reveals that there is newly formed bone growing up to and around the implants. There is no evidence of a fibrosis tissue ring around the implants, also indicating that there is no loosening of the implants. In contrast, the untreated controls remain empty with some evidence of a fibrosis ring around the defect hole. These results indicate good biocompatibility and bioactivity of the converted shell implants. PMID:17684000

  5. Biomimetic hydroxyapatite as a new consolidating agent for archaeological bone

    NASA Astrophysics Data System (ADS)

    North, Alexis E.

    Recent studies on calcareous stone and plaster consolidation have demonstrated considerable potential by bio-mimicking the growth of hydroxyapatite (HAP), the main mineralogical constituent of teeth and bone matrix. These initial conservation applications, together with significant fundamental research on the precipitation of HAP for bioengineering and biomedical applications, offer great promise in the use of HAP as a consolidating agent for archaeological bone and other similar materials such as archaeological teeth, ivory, and antler. Experimental research via the controlled application of diammonium phosphate (DAP) precursors to bone flour, modern bone samples, and archaeological bones, indicated the in situ formation of HAP with a simultaneous increase in the cohesiveness of friable bone material, while preserving the bone's physiochemical properties. These preliminary results point towards a promising new method in archaeological conservation.

  6. Evaluation of anorganic bovine-derived hydroxyapatite matrix/cell binding peptide as a bone graft material in the treatment of human periodontal infrabony defects: A clinico-radiographic study

    PubMed Central

    Fatima, Ghousia; Shivamurthy, Ravindra; Thakur, Srinath; Baseer, Mohammad Abdul

    2015-01-01

    Background: Various bone graft materials have been used in the treatment of periodontal defects. A synthetic bone substitute material composed of P-15 with anorganic bone mineral has been scantly studied. Hence, the present study was aimed to evaluate and compare the efficacy of anorganic bovine-derived hydroxyapatite matrix (ABM)/cell binding peptide (P-15) in human periodontal infrabony defects with that of open flap debridement (OFD) alone. Materials and Methods: A split-mouth, randomized controlled clinical study was designed to investigate the efficacy of ABM/P-15. In this clinical trial, 10 patients having bilateral periodontal infrabony defects were treated either with ABM/P-15 or OFD and followed for a period of 9 months. At baseline and at 9 months probing pocket depth (PPD), relative attachment level (RAL), depth of a defect, and radiographic bone level were measured; and compared between test and control sites. Results: A statistically significant reduction (P < 0.001) in PPD was observed in test sites compared to control sites. Both sites showed a gain in RAL without any significant difference. Similarly, the radiographic evaluation revealed significantly higher radiographic defect fill in test sites as compared to control sites (P < 0.001). Conclusion: ABM/P-15 bone graft material appears to be useful and beneficial in the treatment of human periodontal infrabony defects. PMID:26941516

  7. Bone response to 3D periodic hydroxyapatite scaffolds with and without tailored microporosity to deliver bone morphogenetic protein 2.

    PubMed

    Dellinger, Jennifer G; Eurell, Jo Ann C; Stewart, Matthew; Jamison, Russell D

    2006-02-01

    Three types of model hydroxyapatite (HA) scaffolds were implanted in the metacarpal and metatarsal bones of goats. Scaffolds, consisting of a latticed pattern of rods, were fabricated with a solid freeform fabrication (SFF) technique. All scaffolds contained macropores; some were also fabricated with micropores (5.2 +/- 2.0 microm). Recombinant human bone morphogenetic protein-2 (rhBMP-2) was added to some microporous scaffolds. rhBMP-2 caused increased percent filled with bone tissue compared to microporous scaffolds without rhBMP-2. Lamellar bone in the scaffolds was aligned perpendicular to the long axis of the bone near the junctions of the rods that make up the scaffold but was more random away from the junctions of rods. Microporous scaffolds stained beneath areas of contact with new bone. This staining might indicate either extracellular matrix (ECM) in the rods, byproducts of ECM production, or reaction of cellular products with the scaffold. PMID:16270335

  8. Calcium silicate ceramic scaffolds toughened with hydroxyapatite whiskers for bone tissue engineering

    SciTech Connect

    Feng, Pei; Wei, Pingpin; Li, Pengjian; Gao, Chengde; Shuai, Cijun; Peng, Shuping

    2014-11-15

    Calcium silicate possessed excellent biocompatibility, bioactivity and degradability, while the high brittleness limited its application in load-bearing sites. Hydroxyapatite whiskers ranging from 0 to 30 wt.% were incorporated into the calcium silicate matrix to improve the strength and fracture resistance. Porous scaffolds were fabricated by selective laser sintering. The effects of hydroxyapatite whiskers on the mechanical properties and toughening mechanisms were investigated. The results showed that the scaffolds had a uniform and continuous inner network with the pore size ranging between 0.5 mm and 0.8 mm. The mechanical properties were enhanced with increasing hydroxyapatite whiskers, reached a maximum at 20 wt.% (compressive strength: 27.28 MPa, compressive Young's modulus: 156.2 MPa, flexural strength: 15.64 MPa and fracture toughness: 1.43 MPa·m{sup 1/2}) and then decreased by addition of more hydroxyapatite whiskers. The improvement of mechanical properties was due to whisker pull-out, crack deflection and crack bridging. Moreover, the degradation rate decreased with the increase of hydroxyapatite whisker content. A layer of bone-like apatite was formed on the scaffold surfaces after being soaked in simulated body fluid. Human osteoblast-like MG-63 cells spread well on the scaffolds and proliferated with increasing culture time. These findings suggested that the calcium silicate scaffolds reinforced with hydroxyapatite whiskers showed great potential for bone regeneration and tissue engineering applications. - Highlights: • HA whiskers were incorporated into CS to improve the properties. • The scaffolds were successfully fabricated by SLS. • Toughening mechanisms was whisker pull-out, crack deflection and bridging. • The scaffolds showed excellent apatite forming ability.

  9. Effect of strontium-containing hydroxyapatite bone cement on bone remodeling following hip replacement.

    PubMed

    Ni, Guo X; Lin, Jian H; Chiu, Peter K Y; Li, Zhao Y; Lu, William W

    2010-01-01

    It is uncertain whether the use of bioactive bone cement has any beneficial effect on local bone adaptation following hip replacement. In this study, twelve goats underwent cemented hip hemiarthroplasty unilaterally, with either PMMA bone cement or strontium-containing hydroxyapatite (Sr-HA) bioactive bone cement. Nine months later, the femoral cortical bones at different levels were analyzed by microhardness testing and micro-CT scanning. Extensive bone remodeling was found at proximal and mid-levels in both PMMA and Sr-HA groups. However, with regard to the differences of bone mineral density, cortical bone area and bone hardness between implanted and non-implanted femur, less decreases were found in Sr-HA group than PMMA group at proximal and mid-levels, and significant differences were shown for bone area and hardness at proximal level. The results suggested that the use of Sr-HA cement might alleviate femoral bone remodeling after hip replacement. PMID:19728042

  10. Controlled nucleation of hydroxyapatite on alginate scaffolds for stem cell-based bone tissue engineering

    PubMed Central

    Suárez-González, Darilis; Barnhart, Kara; Saito, Eiji; Vanderby, Ray; Hollister, Scott; Murphy, William L.

    2010-01-01

    Current bone tissue engineering strategies aim to grow a tissue similar to native bone by combining cells and biologically active molecules with a scaffold material. In this study, a macroporous scaffold made from the seaweed-derived polymer alginate was synthesized and mineralized for cell-based bone tissue engineering applications. Nucleation of a bone-like hydroxyapatite mineral was achieved by incubating the scaffold in modified simulated body fluids (mSBF) for four weeks. Analysis using scanning electron microscopy and energy dispersive x-ray analysis indicated growth of a continuous layer of mineral primarily composed of calcium and phosphorous. X-ray diffraction analysis showed peaks associated with hydroxyapatite, the major inorganic constituent of human bone tissue. In addition to the mineral characterization, the ability to control nucleation on the surface, into the bulk of the material, or on the inner pore surfaces of scaffolds was demonstrated. Finally, human MSCs attached and proliferated on the mineralized scaffolds and cell attachment improved when seeding cells on mineral coated alginate scaffolds. This novel alginate- HAP composite material could be used in bone tissue engineering as a scaffold material to deliver cells, and perhaps also biologically active molecules. PMID:20574984

  11. Bone protein extraction without demineralization utilizing principles from hydroxyapatite chromatography

    PubMed Central

    Cleland, Timothy P.; Vashishth, Deepak

    2014-01-01

    Historically, extraction of bone proteins has relied on the use of demineralization to better retrieve proteins from the extracellular matrix; however, demineralization can be a slow process that restricts subsequent analysis of the samples. Here, we developed a novel protein extraction method that does not use demineralization, but utilizes a methodology from hydroxyapatite chromatography where high concentrations of ammonium phosphate and ammonium bicarbonate are used to extract bone proteins. We report that this method has a higher yield than previously published small-scale extant bone extractions, with and without demineralization. Furthermore, after digestion with trypsin and subsequent HPLC-MS/MS analysis, we were able to detect several extracellular matrix and vascular proteins in addition to collagen I and osteocalcin. Our new method has the potential to isolate proteins in a short period (4 hrs) and provide information about bone proteins that may be lost during demineralization or with the use of denaturing agents. PMID:25535955

  12. Biomimetic Nucleation of Hydroxyapatite Crystals Mediated by Antheraea pernyi Silk Sericin Promotes Osteogenic Differentiation of Human Bone Marrow Derived Mesenchymal Stem Cells

    PubMed Central

    2015-01-01

    Biomacromolecules have been used as templates to grow hydroxyapatite crystals (HAps) by biomineralization to fabricate mineralized materials for potential application in bone tissue engineering. Silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation. Mineralization of the silk sericin from Antheraea pernyi (A. pernyi) silkworm has rarely been reported. Here, for the first time, nucleation of HAps on A. pernyi silk sericin (AS) was attempted through a wet precipitation method and consequently the cell viability and osteogenic differentiation of BMSCs on mineralized AS were investigated. It was found that AS mediated the nucleation of HAps in the form of nanoneedles while self-assembling into β-sheet conformation, leading to the formation of a biomineralized protein based biomaterial. The cell viability assay of BMSCs showed that the mineralization of AS stimulated cell adhesion and proliferation, showing that the resultant AS biomaterial is biocompatible. The differentiation assay confirmed that the mineralized AS significantly promoted the osteogenic differentiation of BMSCs when compared to nonmineralized AS as well as other types of sericin (B. mori sericin), suggesting that the resultant mineralized AS biomaterial has potential in promoting bone formation. This result represented the first work proving the osteogenic differentiation of BMSCs directed by silk sericin. Therefore, the biomineralization of A. pernyi silk sericin coupled with seeding BMSCs on the resultant mineralized biomaterials is a useful strategy to develop the potential application of this unexplored silk sericin in the field of bone tissue engineering. This study lays the foundation for the use of A. pernyi silk sericin as a potential scaffold for tissue engineering. PMID:24666022

  13. Pure and Strontium Doped Nano Hydroxyapatite: New Approach for Bone Implant and Drug Delivery System

    NASA Astrophysics Data System (ADS)

    Tank, Kashmira P.; Vasant, Sonal R.; Chudasama, Kiran S.; Thaker, Vrinda S.; Joshi, Mihir J.

    2011-07-01

    Hydroxyapatite, (Ca10(PO4)6(OH)2-Hap), an excellent inorganic biomaterial, find various applications. The chemical composition of Hap is similar to that of the inorganic matrix of human bone and dental enamel. It is also used in drug delivery system and coating of bone implant. In the present study, pure nano Hap and Strontium doped nano-Hap (Sr-Hap) with different concentrations were synthesized by surfactant mediated approach. The samples were characterized by EDAX, XRD and TEM. The hemolytic properties were also studied and it proved that all the samples were non-hemolytic.

  14. Radially and axially graded multizonal bone graft substitutes targeting critical-sized bone defects from polycaprolactone/hydroxyapatite/tricalcium phosphate.

    PubMed

    Ergun, Asli; Yu, Xiaojun; Valdevit, Antonio; Ritter, Arthur; Kalyon, Dilhan M

    2012-12-01

    Repair and regeneration of critical sized defects via the utilization of polymeric bone graft substitutes are challenges. Here, we introduce radially and axially graded multizonal bone graft substitutes fabricated from polycaprolactone (PCL), and PCL biocomposites with osteoconductive particles, that is, hydroxyapatite (HA), and β-tricalcium phosphate (TCP). The novel bone graft substitutes should provide a greater degree of freedom to the orthopedic surgeon especially for repair of critically sized bone defects. The modulus of the graft substitute could be tailored in the axial direction upon the systematic variation of the HA/TCP concentration, while in the radial direction the bone graft substitute consisted of an outer layer with high stiffness, encapsulating a softer core with greater porosity. The biocompatibility of the bone graft substitutes was investigated using in vitro culturing of human bone marrow-derived stromal cells followed by the analysis of cell proliferation and differentiation rates. The characterization of the tissue constructs included the enzymatic alkaline phosphates (ALP) activity, microcomputed tomography imaging, and polymerase chain reaction analysis involving the expressions of bone markers, that is, Runx2, ALP, collagen type I, osteopontin, and osteocalcin, overall demonstrating the differentiation of bone marrow derived stem cells (BMSCs) via osteogenic lineage and formation of mineralized bone tissue. PMID:22764839

  15. Radially and Axially Graded Multizonal Bone Graft Substitutes Targeting Critical-Sized Bone Defects from Polycaprolactone/Hydroxyapatite/Tricalcium Phosphate

    PubMed Central

    Ergun, Asli; Yu, Xiaojun; Valdevit, Antonio; Ritter, Arthur

    2012-01-01

    Repair and regeneration of critical sized defects via the utilization of polymeric bone graft substitutes are challenges. Here, we introduce radially and axially graded multizonal bone graft substitutes fabricated from polycaprolactone (PCL), and PCL biocomposites with osteoconductive particles, that is, hydroxyapatite (HA), and β-tricalcium phosphate (TCP). The novel bone graft substitutes should provide a greater degree of freedom to the orthopedic surgeon especially for repair of critically sized bone defects. The modulus of the graft substitute could be tailored in the axial direction upon the systematic variation of the HA/TCP concentration, while in the radial direction the bone graft substitute consisted of an outer layer with high stiffness, encapsulating a softer core with greater porosity. The biocompatibility of the bone graft substitutes was investigated using in vitro culturing of human bone marrow-derived stromal cells followed by the analysis of cell proliferation and differentiation rates. The characterization of the tissue constructs included the enzymatic alkaline phosphates (ALP) activity, microcomputed tomography imaging, and polymerase chain reaction analysis involving the expressions of bone markers, that is, Runx2, ALP, collagen type I, osteopontin, and osteocalcin, overall demonstrating the differentiation of bone marrow derived stem cells (BMSCs) via osteogenic lineage and formation of mineralized bone tissue. PMID:22764839

  16. Femur bone repair in ovariectomized rats under the local action of alendronate, hydroxyapatite and the association of alendronate and hydroxyapatite

    PubMed Central

    Canettieri, Antonio Carlos Victor; Colombo, Carlos Eduardo Dias; Chin, Chung Man; Faig-Leite, Horácio

    2009-01-01

    An evaluation was made of the local action of alendronate sodium (A), hydroxyapatite (HA) and the association of both substances (A + HA), in different molar concentrations, on the femur bone repair of ovariectomized rats. Ninety-eight animals were divided into seven groups: control (C), starch (S), alendronate 1 mol (A1), alendronate 2 mols (A2), hydroxyapatite 1 mol (HA1), hydroxyapatite 2 mols (HA2) and the association of alendronate + hydroxyapatite (A + HA). Rats weighing about 250 g were ovariectomized and 2.5-mm diameter bone defects were made on the left femur 30 days later. Each experimental group had defects filled with appropriate material, except for group C (control). The animals were killed 7 and 21 days after surgery. Histological, histomorphometric and statistical analyses of bone neoformation in the bone defect site were performed. From the histological standpoint, the major differences occurred after 21 days. All specimens in groups C, S, HA1 and HA2 presented linear closure of the bone defect, and most animals in groups A1, A2 and A + HA showed no bone neoformation in the central area of the defect. No statistically significant difference was found among the experimental groups after 7 days; after 21 days, group HA2 presented the highest amount of neoformed bone. There was no significant difference among groups A1, A2 and A + HA in the two study periods. It was concluded that alendronate, either isolated or in association with hydroxyapatite, had an adverse effect on bone repair in this experimental model. Moreover, the hydroxyapatite used here proved to be biocompatible and osteoconductive, with group HA2 showing the best results. PMID:19765106

  17. Bone properties surrounding hydroxyapatite-coated custom osseous integrated dental implants.

    PubMed

    Baker, M I; Eberhardt, A W; Martin, D M; McGwin, G; Lemons, J E

    2010-10-01

    Calcium phosphate (hydroxyapatite or HA) coatings have been applied to Custom Osseous Integrated Implants (COIIs) to improve the quality of the bone-implant integration, yet little is known concerning the biomechanical properties of bone surrounding the HA-coated implants in humans over the long term. The purpose of this study was to characterize the mechanical and histomorphometric properties of the bone along the implant interface. Specimens were prepared from three similar mandibular implants that were functional in three female patients for about 11 years. Histomorphometric analyses showed bone-implant contact averaging 75% for all specimens. Area coverage of residual HA-coating ranged from 52 to 70%. When compared with previous studies, these results show a relatively high percentage of residual HA after a decade in vivo. Nanoindentation showed similar average values of hardness and modulus (p = 0.53 and p = 0.56, respectively) comparing bone adjacent to residual HA-coating and regions where the coating was absent. The elastic modulus was significantly lower for bone near the bone-implant interface (<200 μm) as compared with bone distant (>1000 μm) from the interface (p = 0.05), thereby reflecting different properties of the bone near these interfaces. Backscattered electron imaging showed darker gray levels which indicated decreased mineral content in bone adjacent to the implant, consistent with the nanoindentation results. PMID:20725958

  18. Hybrid Hydroxyapatite Nanoparticle Colloidal Gels are Injectable Fillers for Bone Tissue Engineering

    PubMed Central

    Gu, Zhen; Jamal, Syed; Detamore, Michael S.

    2013-01-01

    Injectable bone fillers have emerged as an alternative to the invasive surgery often required to treat bone defects. Current bone fillers may benefit from improvements in dynamic properties such as shear thinning during injection and recovery of material stiffness after placement. Negatively charged inorganic hydroxyapatite (HAp) nanoparticles (NPs) were assembled with positively charged organic poly(d,l-lactic-co-glycolic acid) (PLGA) NPs to create a cohesive colloidal gel. This material is held together by electrostatic forces that may be disrupted by shear to facilitate extrusion, molding, or injection. Scanning electron micrographs of the dried colloidal gels showed a well-organized, three-dimensional porous structure. Rheology tests revealed that certain colloidal gels could recover after being sheared. Human umbilical cord mesenchymal stem cells were also highly viable when seeded on the colloidal gels. HAp/PLGA NP colloidal gels offer an attractive scheme for injectable filling and regeneration of bone tissue. PMID:23815275

  19. PCL-coated hydroxyapatite scaffold derived from cuttlefish bone: morphology, mechanical properties and bioactivity.

    PubMed

    Milovac, Dajana; Gallego Ferrer, Gloria; Ivankovic, Marica; Ivankovic, Hrvoje

    2014-01-01

    In the present study, poly(ε-caprolactone)-coated hydroxyapatite scaffold derived from cuttlefish bone was prepared. Hydrothermal transformation of aragonitic cuttlefish bone into hydroxyapatite (HAp) was performed at 200°C retaining the cuttlebone architecture. The HAp scaffold was coated with a poly(ε-caprolactone) (PCL) using vacuum impregnation technique. The compositional and morphological properties of HAp and PCL-coated HAp scaffolds were studied by means of X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM) with energy dispersive X-ray (EDX) analysis. Bioactivity was tested by immersion in Hank's balanced salt solution (HBSS) and mechanical tests were performed at compression. The results showed that PCL-coated HAp (HAp/PCL) scaffold resulted in a material with improved mechanical properties that keep the original interconnected porous structure indispensable for tissue growth and vascularization. The compressive strength (0.88MPa) and the elastic modulus (15.5MPa) are within the lower range of properties reported for human trabecular bones. The in vitro mineralization of calcium phosphate (CP) that produces the bone-like apatite was observed on both the pure HAp scaffold and the HAp/PCL composite scaffold. The prepared bioactive scaffold with enhanced mechanical properties is a good candidate for bone tissue engineering applications. PMID:24268280

  20. Hydroxyapatite-Based Biomaterials vs. Autologous Bone Graft in Spinal Fusion: An in Vivo Animal Study.

    PubMed

    Bròdano, Giovanni Barbanti; Giavaresi, Gianluca; Lolli, Francesco; Salamanna, Francesca; Parrilli, Annapaola; Martini, Lucia; Griffoni, Cristiana; Greggi, Tiziana; Arcangeli, Elena; Pressato, Daniele; Boriani, Stefano; Fini, Milena

    2014-04-01

    Study Design. An in vivo study was designed to compare the efficacy of biomimetic Magnesium-Hydroxyapatite (MgHA) and of human demineralised bone matrix (HDBM), both dispersed in a mixture of biomimetic Mg-HA nanoparticles, with that of an autologous bone graft.Objective. The objective of this study was to evaluate two new bone substitutes as alternatives to a bone autograft for spinal fusion, determining their osteoinductive and osteoconductive properties, and their capacity of remodeling, using a large animal model.Summary of Background Data. Spinal fusion is a common surgical procedure and it is performed for different conditions. A successful fusion requires potentially osteogenic, osteoinductive and osteoconductive biomaterials.Methods. A posterolateral spinal fusion model involved 18 sheep, bilaterally implanting test materials between the vertebral transverse processes. The animals were divided into two groups: one fusion level was treated with MgHA (Group 1) or with HDBM-MgHA (Group 2). The other fusion level received bone autografts in both groups.Results. Radiographic, histological and microtomographic results indicated good osteointegration between the spinous process and the vertebral foramen for both materials. Histomorphometry revealed no significant differences between MgHA and autologous bone for all the parameters examined while significantly lower values of bone volume were observed between HDBM-MgHA and autologous bone. Moreover, the normalisation of the histomorphometrical data with autologous bone revealed that MgHA showed a significantly higher value of bone volume and a lower value of trabecular number, more similar to autologous bone, in comparison to HDBM-MgHA.Conclusion. The study showed that the use of MgHA in an ovine model of spinal fusion led to the deposition of new bone tissue without qualitative and quantitative differences with respect to new bone formed with autologous bone while the HDBM-MgHA led to a reduced deposition of newly

  1. Hydroxyapatite whisker-reinforced polyetherketoneketone bone ingrowth scaffolds.

    PubMed

    Converse, Gabriel L; Conrad, Timothy L; Merrill, Christina H; Roeder, Ryan K

    2010-03-01

    Hydroxyapatite (HA) whisker-reinforced polyetherketoneketone (PEKK) bone ingrowth scaffolds were prepared and characterized. High levels of porosity (75-90%) and HA whisker reinforcement (0-40 vol.%) were attained using a powder processing approach to mix the HA whiskers, PEKK powder and a NaCl porogen, followed by compression molding at 350-375 degrees Celsius and particle leaching to remove the porogen. The scaffold architecture and microstructure exhibited characteristics known to be favorable for osteointegration. Scaffold porosity was interconnected with a mean pore size in the range 200-300 microm as measured by micro-computed tomography. HA whiskers were embedded within and exposed on the surface of scaffold struts, producing a microscale surface topography, shown by von Kossa staining and scanning electron microscopy. Therefore, HA whisker-reinforced PEKK bone ingrowth scaffolds may be advantageous for orthopedic implant fixation, including interbody spinal fusion. PMID:19665061

  2. Nano hydroxyapatite-coated implants improve bone nanomechanical properties.

    PubMed

    Jimbo, R; Coelho, P G; Bryington, M; Baldassarri, M; Tovar, N; Currie, F; Hayashi, M; Janal, M N; Andersson, M; Ono, D; Vandeweghe, S; Wennerberg, A

    2012-12-01

    Nanostructure modification of dental implants has long been sought as a means to improve osseointegration through enhanced biomimicry of host structures. Several methods have been proposed and demonstrated for creating nanotopographic features; here we describe a nanoscale hydroxyapatite (HA)-coated implant surface and hypothesize that it will hasten osseointegration and improve its quality relative to that of non-coated implants. Twenty threaded titanium alloy implants, half prepared with a stable HA nanoparticle surface and half grit-blasted, acid-etched, and heat-treated (HT), were inserted into rabbit femurs. Pre-operatively, the implants were morphologically and topographically characterized. After 3 weeks of healing, the samples were retrieved for histomorphometry. The nanomechanical properties of the surrounding bone were evaluated by nanoindentation. While both implants revealed similar bone-to-implant contact, the nanoindentation demonstrated that the tissue quality was significantly enhanced around the HA-coated implants, validating the postulated hypothesis. PMID:23045363

  3. Hydroxyapatite-titanium bulk composites for bone tissue engineering applications.

    PubMed

    Kumar, Alok; Biswas, Krishanu; Basu, Bikramjit

    2015-02-01

    The research work on bulk hydroxyapatite (HA)-based composites are driven by the need to develop biomaterials with better mechanical properties without compromising its bioactivity and biocompatibility properties. Despite several years of research, the mechanical properties of the HA-based composites still need to be enhanced to match the properties of natural cortical bone. In this regard, the scope of this review on the HA-based bulk biomaterials is limited to the processing and the mechanical as well as biocompatibility properties for bone tissue engineering applications of a model system that is hydroxyapatite-titanium (HA-Ti) bulk composites. It will be discussed in this review how HA-Ti based bulk composites can be processed to have better fracture toughness and strength without compromising biocompatibility. The advantages of the functionally gradient materials to integrate the mechanical and biocompatibility properties is a promising approach in hard tissue engineering and has been emphasized here in reference to the limited literature reports. On the biomaterials fabrication aspect, the recent results are discussed to demonstrate that advanced manufacturing techniques, like spark plasma sintering can be adopted as a processing route to restrict the sintering reactions, while enhancing the mechanical properties. Various toughening mechanisms related to careful tailoring of microstructure are discussed. The in vitro cytocompatibilty, cell fate processes as well as in vivo biocompatibility results are also reviewed and the use of flow cytometry to quantify in vitro cell fate processes is being emphasized. PMID:24737723

  4. Enhanced initial bone regeneration with inorganic polyphosphate-adsorbed hydroxyapatite.

    PubMed

    Morita, K; Doi, K; Kubo, T; Takeshita, R; Kato, S; Shiba, T; Akagawa, Y

    2010-07-01

    Inorganic polyphosphate (poly(P)) can promote binding between fibroblast growth factors and their receptors and enhance osteoblastic cell differentiation and calcification. This study evaluated the possibilities for poly(P) adsorbed onto interconnected porous calcium hydroxyapatite (IP-CHA) as a new bone regeneration material. Prepared 1%, 5%, 25% and 50% poly(P)/IP-CHA composites showed the elution peak of poly(P) between 15 and 20 min, respectively, with the highest value from 50% poly(P)/IP-CHA in vitro. Histologically, at 1 week of placement into the femur of rabbits, granulation tissue had penetrated into the pores in all composites and IP-CHA as a control. In contrast, at 2 weeks of placement, newly formed lamellar bone was found in all groups, although a higher amount of bone regeneration was obviously formed in the 25% and 50% poly(P)/IP-CHA with a significantly higher value of bone regeneration ratio of 50% poly(P)/IP-CHA. These results indicate that 25% and 50% poly(P)/IP-CHA composites may enhance initial bone regeneration. PMID:20056175

  5. Bacterial Cellulose-Hydroxyapatite Nanocomposites for Bone Regeneration

    PubMed Central

    Saska, S.; Barud, H. S.; Gaspar, A. M. M.; Marchetto, R.; Ribeiro, S. J. L.; Messaddeq, Y.

    2011-01-01

    The aim of this study was to develop and to evaluate the biological properties of bacterial cellulose-hydroxyapatite (BC-HA) nanocomposite membranes for bone regeneration. Nanocomposites were prepared from bacterial cellulose membranes sequentially incubated in solutions of CaCl2 followed by Na2HPO4. BC-HA membranes were evaluated in noncritical bone defects in rat tibiae at 1, 4, and 16 weeks. Thermogravimetric analyses showed that the amount of the mineral phase was 40%–50% of the total weight. Spectroscopy, electronic microscopy/energy dispersive X-ray analyses, and X-ray diffraction showed formation of HA crystals on BC nanofibres. Low crystallinity HA crystals presented Ca/P a molar ratio of 1.5 (calcium-deficient HA), similar to physiological bone. Fourier transformed infrared spectroscopy analysis showed bands assigned to phosphate and carbonate ions. In vivo tests showed no inflammatory reaction after 1 week. After 4 weeks, defects were observed to be completely filled in by new bone tissue. The BC-HA membranes were effective for bone regeneration. PMID:21961004

  6. Synthesis and characterization of hydroxyapatite from fish bone waste

    NASA Astrophysics Data System (ADS)

    Marliana, Ana; Fitriani, Eka; Ramadhan, Fauzan; Suhandono, Steven; Yuliani, Keti; Windarti, Tri

    2015-12-01

    Waste fish bones is a problem stemming from activities in the field of fisheries and it has not been used optimally. Fish bones contain calcium as natural source that used to synthesize hydroxyapatite (HA). In this research, HA synthesized from waste fish bones as local wisdom in Semarang. The goal are to produce HA with cheaper production costs and to reduce the environmental problems caused by waste bones. The novelty of this study was using of local fish bone as a source of calcium and simple method of synthesis. Synthesis process of HA can be done through a maceration process with firing temperatures of 1000°C or followed by a sol-gel method with firing at 550°C. The results are analyzed using FTIR (Fourier Transform Infrared), XRD (X-Ray Diffraction) and SEM-EDX (Scanning Electron Microscopy-Energy Dispersive X-Ray). FTIR spectra showed absorption of phosphate and OH group belonging to HA as evidenced by the results of XRD. The average grain size by maceration and synthesized results are not significant different, which is about 69 nm. The ratio of Ca/P of HA by maceration result is 0.89, then increase after continued in the sol-gel process to 1.41. Morphology of HA by maceration results are regular and uniform particle growth, while the morphology of HA after the sol-gel process are irregular and agglomerated.

  7. Synthesis and characterization of hydroxyapatite from fish bone waste

    SciTech Connect

    Marliana, Ana Fitriani, Eka; Ramadhan, Fauzan; Suhandono, Steven; Yuliani, Keti; Windarti, Tri

    2015-12-29

    Waste fish bones is a problem stemming from activities in the field of fisheries and it has not been used optimally. Fish bones contain calcium as natural source that used to synthesize hydroxyapatite (HA). In this research, HA synthesized from waste fish bones as local wisdom in Semarang. The goal are to produce HA with cheaper production costs and to reduce the environmental problems caused by waste bones. The novelty of this study was using of local fish bone as a source of calcium and simple method of synthesis. Synthesis process of HA can be done through a maceration process with firing temperatures of 1000°C or followed by a sol-gel method with firing at 550°C. The results are analyzed using FTIR (Fourier Transform Infrared), XRD (X-Ray Diffraction) and SEM-EDX (Scanning Electron Microscopy-Energy Dispersive X-Ray). FTIR spectra showed absorption of phosphate and OH group belonging to HA as evidenced by the results of XRD. The average grain size by maceration and synthesized results are not significant different, which is about 69 nm. The ratio of Ca/P of HA by maceration result is 0.89, then increase after continued in the sol-gel process to 1.41. Morphology of HA by maceration results are regular and uniform particle growth, while the morphology of HA after the sol-gel process are irregular and agglomerated.

  8. Effects of fluoridation of porcine hydroxyapatite on osteoblastic activity of human MG63 cells

    NASA Astrophysics Data System (ADS)

    Li, Zhipeng; Huang, Baoxin; Mai, Sui; Wu, Xiayi; Zhang, Hanqing; Qiao, Wei; Luo, Xin; Chen, Zhuofan

    2015-06-01

    Biological hydroxyapatite, derived from animal bones, is the most widely used bone substitute in orthopedic and dental treatments. Fluorine is the trace element involved in bone remodeling and has been confirmed to promote osteogenesis when administered at the appropriate dose. To take advantage of this knowledge, fluorinated porcine hydroxyapatite (FPHA) incorporating increasing levels of fluoride was derived from cancellous porcine bone through straightforward chemical and thermal treatments. Physiochemical characteristics, including crystalline phases, functional groups and dissolution behavior, were investigated on this novel FPHA. Human osteoblast-like MG63 cells were cultured on the FPHA to examine cell attachment, cytoskeleton, proliferation and osteoblastic differentiation for in vitro cellular evaluation. Results suggest that fluoride ions released from the FPHA play a significant role in stimulating osteoblastic activity in vitro, and appropriate level of fluoridation (1.5 to 3.1 atomic percents of fluorine) for the FPHA could be selected with high potential for use as a bone substitute.

  9. Effect of Hydroxyapatite on Bone Integration in a Rabbit Tibial Defect Model

    PubMed Central

    Sohn, Sung-Keun; Kim, Kyung-Taek; Kim, Chul-Hong; Ahn, Hee-Bae; Rho, Mee-Sook; Jeong, Min-Ho; Sun, Sang-Kyu

    2010-01-01

    Background The aim of the present study was to prepare hydroxyapatite (HA) and then characterize its effect on bone integration in a rabbit tibial defect model. The bone formation with different designs of HA was compared and the bony integration of several graft materials was investigated qualitatively by radiologic and histologic study. Methods Ten rabbits were included in this study; two holes were drilled bilaterally across the near cortex and the four holes in each rabbit were divided into four treatment groups (HAP, hydroxyapatite powder; HAC, hydroxyapatite cylinder; HA/TCP, hydroxyapatite/tri-calcium phosphate cylinder, and titanium cylinder). The volume of bone ingrowth and the change of bone mineral density were statistically calculated by computed tomography five times for each treatment group at 0, 2, 4, 6, and 8 weeks after grafting. Histologic analysis was performed at 8 weeks after grafting. Results The HAP group showed the most pronounced effect on the bone ingrowth surface area, which seen at 4, 6, and 8 weeks after graft (p < 0.05). On comparing the change of bone mineral density the bone ingrowth surface area among the 4 groups, there were no statistically significant differences among the groups found for any period (p > 0.05). On histological examination, the HAP group revealed well-recovered cortical bone, but the bone was irregularly thickened and haphazardly admixed with powder. The HAC group showed similar histological features to those of the HA/TCP group; the cortical surface of the newly developed bone was smooth and the bone matrix on the surface of the cylinder was regularly arranged. Conclusions We concluded that both the hydroxyapatite powder and cylinder models investigated in our study may be suitable as a bone substitute in the rabbit tibial defect model, but their characteristic properties are quite different. In contrast to hydroxyapatite powder, which showed better results for the bone ingrowth surface, the hydroxyapatite

  10. Dissolution of human teeth-derived hydroxyapatite.

    PubMed

    Seo, Dong Seok; Lee, Jong Kook

    2008-01-01

    We have been interested in human teeth which consist of hydroxyapatite (HA), but do not degrade for a long time. In order to overcome dissolution and mechanical degradation of man-made HA, biologically derived hydroxyapatite (BHA) ceramics were prepared from human teeth and their dissolving behavior was investigated in distilled water for 3-14 days and compared with an artificial HA made of synthetic HA powder. BHA ceramics were prepared by calcining freshly extracted human teeth at 900 degrees C and followed by sintering at 1200 degrees C for 2 h. All detectable peaks in the artificial HA are identical to HA lattice planes, whereas BHA consisted of a mixture of HA and beta-tricalcium phosphate (TCP). Although the artificial HA was expected to be stable in water, the surface dissolution initiated at grain boundaries followed by generated many separated grains and their associated pores. On the other hand, BHA showed that definite grains considered as beta-TCP were predominantly dissolved and the grains were separated from the matrix leaving pores. In the mean time, the rest region, mainly consisting of HA, did not show any evidence of dissolution. It indicates that BHA showed rather stable grain boundaries and lack of excessive dissolution in liquid environment. PMID:17943445

  11. The use of demineralized freeze-dried bone and nonresorbable hydroxyapatite in the treatment of buccal dehiscences around implants: clinical reports.

    PubMed

    Fugazzotto, P A; Hempton, T J

    1993-01-01

    A mixture of equal parts of demineralized freeze-dried human cortical bone and nonresorbable porous hydroxyapatite was used in conjunction with expanded polytetrafluoroethylene membrane to promote deposition of bone in buccal dehiscences around dental implants. Two case studies are presented. PMID:8358377

  12. A long term histological analysis of effect of interposed hydroxyapatite between bone and bone cement in THA and TKA.

    PubMed

    Oonishi, Hironobu

    2012-01-01

    The standard cementing technique for total hip and knee arthroplasty (THA, TKA) was modified by interposing osteoconductive porous polycrystalline hydroxyapatite (HA) granules at the cement-bone interface to augment cement-bone bonding. Twenty-one specimens from the acetabulum and two specimens from the femur containing well-fixed bone-cement interface were retrieved five months to twenty-two years after THA and TKA with an interface bioactive bone cement technique. Histological findings were evaluated in terms of the retrieved sites, the interval between index operation and revision, patient age at retrieval and bone pathology. Dense bone ingrowth was observed in all specimens retrieved from the superior wall of the acetabulum. Cancellous bone ingrowth was observed in specimens retrieved from the infero-medial wall of the acetabulum. Connective tissue interposition and osteolysis were observed at the sites where hydroxyapatite granules were absent. Findings of bone ingrowth for the HA interposed retrievals were similar regardless of the interval between index operation and revision, patient age at retrieval, and bone pathology. Longer survivability of the bone-cement bonding was attributed more to the interposed HA granules. In spite of various degrees of aging, postmenopausal osteoporosis, and pathological bone condition the histological findings were not affected by the bone pathology. There was long-term direct contact between the bone and hydroxyapatite granules. Thus, the interface bioactive bone cement technique can ensure longevity of bone-cement bonding even after the onset of osteoporosis and for pathological bone conditions. PMID:23428252

  13. Silicate-doped hydroxyapatite and its promotive effect on bone mineralization

    NASA Astrophysics Data System (ADS)

    Qiu, Zhi-Ye; Noh, In-Sup; Zhang, Sheng-Min

    2013-03-01

    Bone defect is one of the most common diseases in clinic. Existing therapeutic approaches have encountered many problems, such as lack of autogenous allogeneic bone and immunological rejection to allogeneic implant. Synthetic hydroxyapatite (HA) provided solutions for bone repair, since the HA is the main inorganic component of animals' bone. However, HA has good biocompatibility, but does not possess osteogenic capability, which is of significance for modern bone repair materials. Si is an essential trace element in bone tissue, and it has been demonstrated to be able to promote bone formation. Therefore, silicate-doped hydroxyapatite (Si-HA) may serve as a promising material for bone repair, and promote bone regeneration in the repair. The current review discusses development of Si-HA, focusing on its preparation and characterization, in vitro and in vivo evaluations of the material, positive effect of Si-HA on promoting bone formation in clinical applications, and molecular mechanism investigation of such promotive effect.

  14. A Biphasic Calcium Sulphate/Hydroxyapatite Carrier Containing Bone Morphogenic Protein-2 and Zoledronic Acid Generates Bone

    PubMed Central

    Raina, Deepak Bushan; Isaksson, Hanna; Hettwer, Werner; Kumar, Ashok; Lidgren, Lars; Tägil, Magnus

    2016-01-01

    In orthopedic surgery, large amount of diseased or injured bone routinely needs to be replaced. Autografts are mainly used but their availability is limited. Commercially available bone substitutes allow bone ingrowth but lack the capacity to induce bone formation. Thus, off-the-shelf osteoinductive bone substitutes that can replace bone grafts are required. We tested the carrier properties of a biphasic, calcium sulphate and hydroxyapatite ceramic material, containing a combination of recombinant human bone morphogenic protein-2 (rhBMP-2) to induce bone, and zoledronic acid (ZA) to delay early resorption. In-vitro, the biphasic material released 90% of rhBMP-2 and 10% of ZA in the first week. No major changes were found in the surface structure using scanning electron microscopy (SEM) or in the mechanical properties after adding rhBMP-2 or ZA. In-vivo bone formation was studied in an abdominal muscle pouch model in rats (n = 6/group). The mineralized volume was significantly higher when the biphasic material was combined with both rhBMP-2 and ZA (21.4 ± 5.5 mm3) as compared to rhBMP-2 alone (10.9 ± 2.1 mm3) when analyzed using micro computed tomography (μ-CT) (p < 0.01). In the clinical setting, the biphasic material combined with both rhBMP-2 and ZA can potentially regenerate large volumes of bone. PMID:27189411

  15. A Biphasic Calcium Sulphate/Hydroxyapatite Carrier Containing Bone Morphogenic Protein-2 and Zoledronic Acid Generates Bone.

    PubMed

    Raina, Deepak Bushan; Isaksson, Hanna; Hettwer, Werner; Kumar, Ashok; Lidgren, Lars; Tägil, Magnus

    2016-01-01

    In orthopedic surgery, large amount of diseased or injured bone routinely needs to be replaced. Autografts are mainly used but their availability is limited. Commercially available bone substitutes allow bone ingrowth but lack the capacity to induce bone formation. Thus, off-the-shelf osteoinductive bone substitutes that can replace bone grafts are required. We tested the carrier properties of a biphasic, calcium sulphate and hydroxyapatite ceramic material, containing a combination of recombinant human bone morphogenic protein-2 (rhBMP-2) to induce bone, and zoledronic acid (ZA) to delay early resorption. In-vitro, the biphasic material released 90% of rhBMP-2 and 10% of ZA in the first week. No major changes were found in the surface structure using scanning electron microscopy (SEM) or in the mechanical properties after adding rhBMP-2 or ZA. In-vivo bone formation was studied in an abdominal muscle pouch model in rats (n = 6/group). The mineralized volume was significantly higher when the biphasic material was combined with both rhBMP-2 and ZA (21.4 ± 5.5 mm(3)) as compared to rhBMP-2 alone (10.9 ± 2.1 mm(3)) when analyzed using micro computed tomography (μ-CT) (p < 0.01). In the clinical setting, the biphasic material combined with both rhBMP-2 and ZA can potentially regenerate large volumes of bone. PMID:27189411

  16. Effect of hydroxyapatite-based biomaterials on human osteoblast phenotype.

    PubMed

    Trombelli, L; Penolazzi, L; Torreggiani, E; Farina, R; Lambertini, E; Vecchiatini, R; Piva, R

    2010-03-01

    The present study evaluated human primary osteoblasts and two different osteoblast-like cell lines behaviour when cultured in presence of different hydroxyapatite-based (HA) biomaterials (SINTlife-FIN-CERAMICA S.p.a., Faenza, Italy; Bio-Oss, Geistlich Biomaterials, Woulhusen, Switzerland; Biostite-GABA Vebas, San Giuliano Milanese, MI, Italy), focusing attention on the effect of HA/Biostite in terms of modulation of osteoblastic differentiation. Analysis were about adhesion, proliferation and mineralization activity. Runt-related transcription factor 2 (Runx2), Estrogen Receptor alpha (ERalfa) expression and alkaline phosphatase activity (ALP) were measured as osteoblastic differentiation markers. Determination of viable cells was done with MTT colorimetric assay. Scanning electron microscopy (SEM) analysis was performed on biomaterial-treated cells. All hydroxyapatite-based biomaterials didn't affect cells morphology and viability, whereas only presence of HA/Biostite improved cells adhesion, growth and differentiation. Adhesion and spreading of the primary cells on HA/Biostite were the same showed by two different osteoblast-like cell lines. These results have important implications for both tissue-engineered bone grafts and enhancement of HA implants performance, to develop new teeth's supporting structure therapies and replacement. PMID:20357737

  17. Freeze-cast hydroxyapatite scaffolds for bone tissue engineering applications.

    PubMed

    Fu, Qiang; Rahaman, Mohamed N; Dogan, Fatih; Bal, B Sonny

    2008-06-01

    Freeze casting of aqueous suspensions was investigated as a method for preparing porous hydroxyapatite (HA) scaffolds for eventual application to bone tissue engineering. Suspensions of HA particles (10-20 volume percent) were frozen unidirectionally in a cylindrical mold placed on a cold steel substrate (-20 degrees C). After sublimation of the ice, sintering for 3 h at 1350 degrees C produced constructs with dense HA lamellae, with porosity of approximately 50%, and inter-lamellar pore widths of 5-30 microm. These constructs had compressive strengths of 12 +/- 1 MPa and 5 +/- 1 MPa in the directions parallel and perpendicular to the freezing direction, respectively. Manipulation of the microstructure was achieved by modifying the solvent composition of the suspension used for freeze casting. The use of water-glycerol mixtures (20 wt% glycerol) resulted in the production of constructs with finer pores (1-10 microm) and a larger number of dendritic growth connecting the HA lamellae, and higher strength. On the other hand, the use of water-dioxane mixtures (60 wt% dioxane) resulted in a cellular-type microstructure with larger pores (90-110 microm). The mechanical response showed high strain tolerance (5-10% at the maximum stress), high strain for failure (>20%) and sensitivity to the loading rate. The favorable mechanical behavior of the porous constructs, coupled with the ability to modify their microstructure, indicates the potential of the present freeze casting route for the production of porous scaffolds for bone tissue engineering. PMID:18458369

  18. Nanocomposite bone scaffolds based on biodegradable polymers and hydroxyapatite.

    PubMed

    Becker, Johannes; Lu, Lichun; Runge, M Brett; Zeng, Heng; Yaszemski, Michael J; Dadsetan, Mahrokh

    2015-08-01

    In tissue engineering, development of an osteoconductive construct that integrates with host tissue remains a challenge. In this work, the effect of bone-like minerals on maturation of pre-osteoblast cells was investigated using polymer-mineral scaffolds composed of poly(propylene fumarate)-co-poly(caprolactone) (PPF-co-PCL) and nano-sized hydroxyapatite (HA). The HA of varying concentrations was added to an injectable formulation of PPF-co-PCL and the change in thermal and mechanical properties of the scaffolds was evaluated. No change in onset of degradation temperature was observed due to the addition of HA, however compressive and tensile moduli of copolymer changed significantly when HA amounts were increased in composite formulation. The change in mechanical properties of copolymer was found to correlate well to HA concentration in the constructs. Electron microscopy revealed mineral nucleation and a change in surface morphology and the presence of calcium and phosphate on surfaces was confirmed using energy dispersive X-ray analysis. To characterize the effect of mineral on attachment and maturation of pre-osteoblasts, W20-17 cells were seeded on HA/copolymer composites. We demonstrated that cells attached more to the surface of HA containing copolymers and their proliferation rate was significantly increased. Thus, these findings suggest that HA/PPF-co-PCL composite scaffolds are capable of inducing maturation of pre-osteoblasts and have the potential for use as scaffold in bone tissue engineering. PMID:25504776

  19. Magnetic responsive hydroxyapatite composite scaffolds construction for bone defect reparation

    PubMed Central

    Zeng, Xiao Bo; Hu, Hao; Xie, Li Qin; Lan, Fang; Jiang, Wen; Wu, Yao; Gu, Zhong Wei

    2012-01-01

    Introduction In recent years, interest in magnetic biomimetic scaffolds for tissue engineering has increased considerably. A type of magnetic scaffold composed of magnetic nanoparticles (MNPs) and hydroxyapatite (HA) for bone repair has been developed by our research group. Aim and methods In this study, to investigate the influence of the MNP content (in the scaffolds) on the cell behaviors and the interactions between the magnetic scaffold and the exterior magnetic field, a series of MNP-HA magnetic scaffolds with different MNP contents (from 0.2% to 2%) were fabricated by immersing HA scaffold into MNP colloid. ROS 17/2.8 and MC3T3-E1 cells were cultured on the scaffolds in vitro, with and without an exterior magnetic field, respectively. The cell adhesion, proliferation and differentiation were evaluated via scanning electron microscopy; confocal laser scanning microscopy; and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), alkaline phosphatase, and bone gla protein activity tests. Results The results demonstrated the positive influence of the magnetic scaffolds on cell adhesion, proliferation, and differentiation. Further, a higher amount of MNPs on the magnetic scaffolds led to more significant stimulation. Conclusion The magnetic scaffold can respond to the exterior magnetic field and engender some synergistic effect to intensify the stimulating effect of a magnetic field to the proliferation and differentiation of cells. PMID:22848165

  20. Drug-loaded porous spherical hydroxyapatite granules for bone regeneration.

    PubMed

    Hong, Min-Ho; Son, Jun-Sik; Kim, Kwang-Mahn; Han, Myungho; Oh, Daniel S; Lee, Yong-Keun

    2011-02-01

    Porous spherical hydroxyapatite (HAp) granules, which are not only can be used for bone void filler, but also drug delivery systems, were prepared using a liquid nitrogen method. Various pore and channel structures of spherical granules were obtained by adjusting the ratio of water to HAp powder and the amount of sodium chloride (NaCl). By using the water to powder ratio at 2.0 ml/g and the amount of NaCl at 15 wt% by powder, the spherical granules have optimal pore volume, micro-channel structure and strength to handle as well as the ability to work as a drug delivery system. When the NaCl content was 15 wt%, the micro-channel structure was changed, but the pore volume was maintained. For the drug release test, dexamathasone (Dex) was loaded as a model drug on the prepared HAp granules by the immersion method, and the drug release behavior was curved by a UV/vis spectrophotometer. As a result, different drug release behavior was observed according to micro-channel structural differences. Therefore, it was concluded that the NACl could be applied as the pore and micro-channel structure control agent. Porous spherical HAp granules, which were fabricated by a liquid nitrogen method, show potential as bone void filler with the ability of controlled drug release. PMID:21222142

  1. Cytocompatibility and biocompatibility of nanostructured carbonated hydroxyapatite spheres for bone repair

    PubMed Central

    CALASANS-MAIA, Mônica Diuana; de MELO, Bruno Raposo; ALVES, Adriana Terezinha Neves Novellino; RESENDE, Rodrigo Figueiredo de Brito; LOURO, Rafael Seabra; SARTORETTO, Suelen Cristina; GRANJEIRO, José Mauro; ALVES, Gutemberg Gomes

    2015-01-01

    ABSTRACT Objective The aim of this study was to investigate the in vitro and in vivo biological responses to nanostructured carbonated hydroxyapatite/calcium alginate (CHA) microspheres used for alveolar bone repair, compared to sintered hydroxyapatite (HA). Material and Methods The maxillary central incisors of 45 Wistar rats were extracted, and the dental sockets were filled with HA, CHA, and blood clot (control group) (n=5/period/group). After 7, 21 and 42 days, the samples of bone with the biomaterials were obtained for histological and histomorphometric analysis, and the plasma levels of RANKL and OPG were determined via immunoassay. Statistical analysis was performed by Two-Way ANOVA with post-hoc Tukey test at 95% level of significance. Results The CHA and HA microspheres were cytocompatible with both human and murine cells on an in vitro assay. Histological analysis showed the time-dependent increase of newly formed bone in control group characterized by an intense osteoblast activity. In HA and CHA groups, the presence of a slight granulation reaction around the spheres was observed after seven days, which was reduced by the 42nd day. A considerable amount of newly formed bone was observed surrounding the CHA spheres and the biomaterials particles at 42-day time point compared with HA. Histomorphometric analysis showed a significant increase of newly formed bone in CHA group compared with HA after 21 and 42 days from surgery, moreover, CHA showed almost 2-fold greater biosorption than HA at 42 days (two-way ANOVA, p<0.05) indicating greater biosorption. An increase in the RANKL/OPG ratio was observed in the CHA group on the 7th day. Conclusion CHA spheres were osteoconductive and presented earlier biosorption, inducing early increases in the levels of proteins involved in resorption. PMID:26814461

  2. Low-Power Ultrasounds as a Tool to Culture Human Osteoblasts inside Cancellous Hydroxyapatite

    PubMed Central

    Fassina, Lorenzo; Saino, Enrica; Cusella De Angelis, Maria Gabriella; Magenes, Giovanni; Benazzo, Francesco; Visai, Livia

    2010-01-01

    Bone graft substitutes and cancellous biomaterials have been widely used to heal critical-size long bone defects due to trauma, tumor resection, and tissue degeneration. In particular, porous hydroxyapatite is widely used in reconstructive bone surgery owing to its biocompatibility. In addition, the in vitro modification of cancellous hydroxyapatite with osteogenic signals enhances the tissue regeneration in vivo, suggesting that the biomaterial modification could play an important role in tissue engineering. In this study, we have followed a tissue-engineering strategy where ultrasonically stimulated SAOS-2 human osteoblasts proliferated and built their extracellular matrix inside a porous hydroxyapatite scaffold. The ultrasonic stimulus had the following parameters: average power equal to 149 mW and frequency of 1.5 MHz. In comparison with control conditions, the ultrasonic stimulus increased the cell proliferation and the surface coating with bone proteins (decorin, osteocalcin, osteopontin, type-I collagen, and type-III collagen). The mechanical stimulus aimed at obtaining a better modification of the biomaterial internal surface in terms of cell colonization and coating with bone matrix. The modified biomaterial could be used, in clinical applications, as an implant for bone repair. PMID:20379359

  3. Ossiculoplasty with hydroxyapatite bone cement: our reconstruction philosophy.

    PubMed

    Gérard, Jean-Marc; De Bie, Gersende; Franceschi, Daniel; Deggouj, Naima; Gersdorff, Michel

    2015-07-01

    The main objective of this study is to analyze results obtained with hydroxyapatite bone cement (HABC) ossiculoplasties. This is a retrospective study of a case series. This study was conducted in an academic hospital and tertiary referral center. A total of 127 ossiculoplasties using HABC were evaluated. Ears were divided into three groups according to procedure: group 1 involved reinforcement of the incudostapedial joint with cement and reconstruction of an incus long process defect with cement. Group 2 involved partial ossicular reconstruction between the stapes and malleus handle with HABC. Group 3 was divided into two subgroups. Group 3B entailed reconstruction of the stapes with a mobile footplate (Austin-Kartush type B = group 3B) and group 3F with a fixed footplate (Austin-Kartush type F = group 3F) using a K-Helix piston (Grace Medical, Memphis, TN, USA) or a classical titanium piston (Kurz, Fuerth, Germany) glued to the incus remnant or malleus handle with cement. Anatomical and pre- and postoperative audiological results were assessed. The mean follow-up was 26 ± 14 months. Percentages of average postoperative air-bone gap ≤ 20 dB were 95, 82.5, 50 and 83.3%, and for air-bone gap ≤ 1 0 dB, 80, 50.9, 16.6 and 50% for groups 1, 2, 3B and 3F, respectively. No complications related to the cement or extrusion occurred. Hearing outcomes also remained stable over time. In our experience, ossiculoplasty with cement provides good and stable functional results, is safe, cost effective, and easy to use. HABC with or without biocompatible ossicular prostheses allows repair of different types of ossicular defects with preservation of the anatomical and physiological ossicular chain, as well as improved stability. Reconstruction of the incus long process or incudostapedial joint defect with cement is preferred over partial ossicular reconstruction. PMID:24615652

  4. Properties of carbon nanotube-dispersed Sr-hydroxyapatite injectable material for bone defects

    PubMed Central

    Raucci, M. G.; Alvarez-Perez, M.; Giugliano, D.; Zeppetelli, S.; Ambrosio, L.

    2016-01-01

    This study concerns the synthesis of gel materials based on carbon nanotubes dispersed strontium-modified hydroxyapatite (Sr-HA) at different compositions obtained by sol–gel technology and their influence on human-bone-marrow-derived mesenchymal stem cells. Furthermore, an evaluation of the influence of nanotubes and Strontium on physico-chemical, morphological, rheological and biological properties of hydroxyapatite gel was also performed. Morphological analysis (scanning electron microscopy) shows a homogeneous distribution of modified nanotubes in the ceramic matrix improving the bioactive properties of materials. The biological investigations proved that Sr-HA/carbon nanotube gel containing 0–20 mol (%) of Sr showed no toxic effect and promote the expression of early and late markers of osteogenic differentiation in cell culture performed in basal medium without osteogenic factors. Finally, the SrHA/carbon nanotube gels could have a good potential application as filler in bone repair and regeneration and may be used in the osteoporotic disease treatment. PMID:26816652

  5. Properties of carbon nanotube-dispersed Sr-hydroxyapatite injectable material for bone defects.

    PubMed

    Raucci, M G; Alvarez-Perez, M; Giugliano, D; Zeppetelli, S; Ambrosio, L

    2016-03-01

    This study concerns the synthesis of gel materials based on carbon nanotubes dispersed strontium-modified hydroxyapatite (Sr-HA) at different compositions obtained by sol-gel technology and their influence on human-bone-marrow-derived mesenchymal stem cells. Furthermore, an evaluation of the influence of nanotubes and Strontium on physico-chemical, morphological, rheological and biological properties of hydroxyapatite gel was also performed. Morphological analysis (scanning electron microscopy) shows a homogeneous distribution of modified nanotubes in the ceramic matrix improving the bioactive properties of materials. The biological investigations proved that Sr-HA/carbon nanotube gel containing 0-20 mol (%) of Sr showed no toxic effect and promote the expression of early and late markers of osteogenic differentiation in cell culture performed in basal medium without osteogenic factors. Finally, the SrHA/carbon nanotube gels could have a good potential application as filler in bone repair and regeneration and may be used in the osteoporotic disease treatment. PMID:26816652

  6. Design, synthesis, and initial evaluation of D-glyceraldehyde crosslinked gelatin-hydroxyapatite as a potential bone graft substitute material

    NASA Astrophysics Data System (ADS)

    Florschutz, Anthony Vatroslav

    Utilization of bone grafts for the treatment of skeletal pathology is a common practice in orthopaedic, craniomaxillofacial, dental, and plastic surgery. Autogenous bone graft is the established archetype but has disadvantages including donor site morbidity, limited supply, and prolonging operative time. In order to avoid these and other issues, bone graft substitute materials are becoming increasingly prevalent among surgeons for reconstructing skeletal defects and arthrodesis applications. Bone graft substitutes are biomaterials, biologics, and guided tissue/bone regenerative devices that can be used alone or in combinations as supplements or alternatives to autogenous bone graft. There is a growing interest and trend to specialize graft substitutes for specific indications and although there is good rationale for this indication-specific approach, the development and utility of a more universal bone graft substitute may provide a better answer for patients and surgeons. The aim of the present research focuses on the design, synthesis, and initial evaluation of D-glyceraldehyde crosslinked gelatin-hydroxyapatite composites for potential use as a bone graft substitutes. After initial establishment of rational material design, gelatinhydroxyapatite scaffolds were fabricated with different gelatin:hydroxyapatite ratios and crosslinking concentrations. The synthesized scaffolds were subsequently evaluated on the basis of their swelling behavior, porosity, density, percent composition, mechanical properties, and morphology and further assessed with respect to cell-biomaterial interaction and biomineralization in vitro. Although none of the materials achieved mechanical properties suitable for structural graft applications, a reproducible material design and synthesis was achieved with properties recognized to facilitate bone formation. Select scaffold formulations as well as a subset of scaffolds loaded with recombinant human bone morphogenetic protein-2 were

  7. Hydroxyapatite crystals as a bone graft substitute in benign lytic lesions of bone

    PubMed Central

    Gupta, Anil Kumar; Kumar, Praganesh; Keshav, Kumar; Singh, Anant

    2015-01-01

    Background: Bone grafts are required to fill a cavity created after curettage of benign lytic lesions of the bone. To avoid the problems associated at donor site with autologous bone graft, we require allograft or bone graft substitutes. We evaluated the healing of lytic lesions after hydroxyapatite (HA) grafting by serial radiographs. Materials and Methods: Forty cases of benign lytic lesions of bone were managed by simple curettage and grafting using HA blocks. Commercially available HA of bovine origin (Surgiwear Ltd., Shahjahanpur, India) was used for this purpose. Mean duration of followup was 34.8 months (range 12–84 months). Mean patient age was 19.05 years (range 3–55 years). Radiological staging of graft incorporation was done as per criteria of Irwin et al. 2001. Results: In our series, two cases were in stage I. A total of 11 cases were in stage II and 27 were in stage III. Graft incorporation was radiologically complete by 15 months. Clinical recovery was observed before radiological healing. The average time taken to return to preoperative function was 3 months. Recurrence was observed in giant cell tumor (n = 3) and chondromyxoid fibroma (n = 1). There was no incidence of graft rejection, collapse, growth plate disturbances or antigenic response. Conclusions: We conclude that calcium HA is biologically acceptable bone graft substitute in the management of benign lytic lesions of bone. PMID:26806973

  8. Characteristics of functionalized nano-hydroxyapatite and internalization by human epithelial cell

    PubMed Central

    2011-01-01

    Hydroxyapatite is the main inorganic component of biological bone and tooth enamel, and synthetic hydroxyapatite has been widely used as biomaterials. In this study, a facile method has been developed for the fabrication of arginine-functionalized and europium-doped hydroxyapatite nanoparticles (Arg-Eu-HAP). The synthesized nanoparticles characterized by transmission electron microscopy, X-ray diffractometry, Fourier transform infrared, and Zeta potential analyzer. Its biological properties with DNA binding, cell toxicity, cell binding and intracellular distribution were tested by agarose gel electrophoresis assay, flow cytometry, and fluorescence microscope and laser scanning confocal microscope. The synthesized Arg-Eu-HAP could effectively bind DNA without any cytotoxicity and be internalized into the cytoplasm and perinuclear of human lung epithelial cells. PMID:22108000

  9. Characteristics of functionalized nano-hydroxyapatite and internalization by human epithelial cell

    NASA Astrophysics Data System (ADS)

    Yan-Zhong, Zhao; Yan-Yan, Huang; Jun, Zhu; Shai-Hong, Zhu; Zhi-You, Li; Ke-Chao, Zhou

    2011-11-01

    Hydroxyapatite is the main inorganic component of biological bone and tooth enamel, and synthetic hydroxyapatite has been widely used as biomaterials. In this study, a facile method has been developed for the fabrication of arginine-functionalized and europium-doped hydroxyapatite nanoparticles (Arg-Eu-HAP). The synthesized nanoparticles characterized by transmission electron microscopy, X-ray diffractometry, Fourier transform infrared, and Zeta potential analyzer. Its biological properties with DNA binding, cell toxicity, cell binding and intracellular distribution were tested by agarose gel electrophoresis assay, flow cytometry, and fluorescence microscope and laser scanning confocal microscope. The synthesized Arg-Eu-HAP could effectively bind DNA without any cytotoxicity and be internalized into the cytoplasm and perinuclear of human lung epithelial cells.

  10. [Hydroxyapatite bone substitute (Ostim) in sinus floor elevation. Maxillary sinus floor augmentation: bone regeneration by means of a nanocrystalline in-phase hydroxyapatite (Ostim)].

    PubMed

    Smeets, Ralf; Grosjean, Maurice B; Jelitte, Gerd; Heiland, Max; Kasaj, Adrian; Riediger, Dieter; Yildirim, Murat; Spiekermann, Hubertus; Maciejewski, Oliver

    2008-01-01

    The range of bone regeneration materials suitable for maxillar bone augmentation has increased steadily in the past few years and there is now a wide variety of materials being used. In the present case report, we analyzed the state of bone regeneration after sinus floor augmentation using a nanocrystalline in-phase synthetic anorganic hydroxyapatite bone grafting material (Ostim). A 60-year-old female patient underwent maxillary sinus floor elevation and the cavity was filled with Ostim three years before. Actually, she presented herself with loosening of the dental implant at position 17, as a result of parafunction. At the time of the insertion of a second implant at position 17, bone samples were taken by using a trepan drilling device from the previously augmented area. These samples were analyzed histologically to determine the extent of bone remodeling around the deposits of Ostim. We found that the Ostim deposits were surrounded largely by woven bone and, in parts, by lamellar bone and had facilitated osteoconductive bone regeneration. The adjacent implant, at position 16, which beared a crown exposed to proper biting forces without parafunction, showed proper clinical and radiological characteristics of complete and firm integration into the area which was also filled with Ostim three years ago. We conclude that the use of the nanocrystalline hydroxyapatite Ostim with its stable volume properties appears to be suitable for maxillary sinus floor augmentation. Furthermore, we even found osteoconductive bone regeneration under Ostim near the site of the loosened implant. PMID:18422056

  11. Preparation and characterization of an iron oxide-hydroxyapatite nanocomposite for potential bone cancer therapy

    PubMed Central

    Sneha, Murugesan; Sundaram, Nachiappan Meenakshi

    2015-01-01

    Recently, multifunctional magnetic nanostructures have been found to have potential applications in biomedical and tissue engineering. Iron oxide nanoparticles are biocompatible and have distinctive magnetic properties that allow their use in vivo for drug delivery and hyperthermia, and as T2 contrast agents for magnetic resonance imaging. Hydroxyapatite is used frequently due to its well-known biocompatibility, bioactivity, and lack of toxicity, so a combination of iron oxide and hydroxyapatite materials could be useful because hydroxyapatite has better bone-bonding ability. In this study, we prepared nanocomposites of iron oxide and hydroxyapatite and analyzed their physicochemical properties. The results suggest that these composites have superparamagnetic as well as biocompatible properties. This type of material architecture would be well suited for bone cancer therapy and other biomedical applications. PMID:26491311

  12. Nonceramic hydroxyapatite bone derivative in sinus augmentation procedures: clinical and histomorphometric observations in 10 consecutive cases.

    PubMed

    Artzi, Zvi; Nemcovsky, Carlos E; Dayan, Dan

    2003-08-01

    A synthetic, nonceramic resorbable hydroxyapatite (R-HA) was applied to augment the subantral area in 10 consecutive sinus lift procedures in humans. Implants were simultaneously placed in eight patients; in the remaining two, where residual bone height was less than 3 mm, a two-stage surgical approach was carried out. The aim of the study was to examine, clinically and histopathologically at 12 months, the healing pattern of these augmented sinuses around the implants. In the simultaneous technique, radiopaque grafted mineral surrounded the implants. In the two-stage technique, R-HA particles filled the augmented site and were confined to the subantral area. At the uncovering phase, all implants (n = 36) were stable, with no clinical bone resorption around the cervix. A 2.5-mm trephine bur was used to collect specimens from the 10 augmented sites at the lateral-deep area. Histologically, new bone formation was evident in all examined cores. R-HA particles were primarily surrounded by newly formed bone, mostly woven bone, in different stages of remodeling. However, in the deep areas of the specimen cores, lamellar bone fragments were also seen. Morphometric measurements showed that the mean bone area of the 10 sites was 28.1% at the lateral/external side and 37.8% at the deep/inward side. Under polarizing microscopy, the mean lamellar:woven bone ratio was 1:7.2 at the lateral side and 1:4.2 at the deep end. Differences were statistically significant. R-HA proved to be a suitable filler material for osseointegrated implants in sinus augmentation procedures, since it showed both biocompatible and osteoconductive properties. PMID:12956482

  13. Characterization of derived natural hydroxyapatite (HAp) obtained from different types of tilapia fish bones and scales

    NASA Astrophysics Data System (ADS)

    Fara, A. N. K. A.; Abdullah, H. Z.

    2015-07-01

    Hydroxyapatite, (HAp), Ca10(PO4)6(OH)2, is recognised as a biomaterial that is widely used for bone implant due to its chemical and structural similarity to the mineral components in human bone and enamel. The elements of HAp are primarily composed of calcium and phosphorus molar ratio of calcium to phosphorous is 1.67 capable to promote bone in-growth into prosthetic implant. Enormous amounts of by-product waste produced from fish factories generated an undesirable environmental impact. Thus, this study was conducted to obtain natural biological HAp from different types of tilapia fish bones and scales from fishery waste. Therefore, fish bones and scales can be as cheap source to produce biological HAp for medical applications. For this purpose, fish bones and scales of tilapia fish were boiled at 100°C to remove adhering meat and other impurities. Later, fish bones and scales were separated into several groups and subjected to different calcination temperatures of 800° C and 900° C for 3h respectively. Afterward, all calcined samples were crushed to form a fine powder. The XRD result revealed the presence of derived Hapfrom the samples powder and were identical with standard Hap. Thermo Gravimetric Analysis was carried out to show the thermal stability of the HAp powder from different types of fish bones and scales. SEM results show porous structure appeared in calcined samples compared to raw samples. The findings are the promising alternative to produce calcium and phosphorus from fishery wastes that beneficial to medical applications.

  14. Biomineralized hydroxyapatite nanoclay composite scaffolds with polycaprolactone for stem cell-based bone tissue engineering.

    PubMed

    Ambre, Avinash H; Katti, Dinesh R; Katti, Kalpana S

    2015-06-01

    Nanoclay modified with unnatural amino acid was used to design a nanoclay-hydroxyapatite (HAP) hybrid by mineralizing HAP in the nanoclay galleries mimicking biomineralization. This hybrid (in situ HAPclay) was used to fabricate polycaprolactone (PCL)/in situ HAPclay films and scaffolds for bone regeneration. Cell culture assays and imaging were used to study interactions between human mesenchymal stem cells (hMSCs) and PCL/in situ HAPclay composites (films and scaffolds). SEM imaging indicated MSC attachment, formation of mineralized extracellular (ECM) on PCL/in situ HAPclay films, and infiltration of MSCs to the interior of PCL/in situ HAPclay scaffolds. Mineralized ECM was formed by MSCs without use of osteogenic supplements. AFM imaging performed on this in vitro generated mineralized ECM on PCL/in situ HAPclay films revealed presence of components (collagen and mineral) of hierarchical organization reminiscent of natural bone. Cellular events observed during two-stage seeding experiments on PCL/in situ HAPclay films indicated similarities with events occurring during in vivo bone formation. PCL/in situ HAPclay films showed significantly increased (100-595% increase in elastic moduli) nanomechanical properties and PCL/in situ HAPclay scaffolds showed increased degradation. This work puts forth PCL/in situ HAPclay composites as viable biomaterials for bone tissue engineering. PMID:25331212

  15. Mechanical properties and osteogenic potential of hydroxyapatite-PLGA-collagen biomaterial for bone regeneration.

    PubMed

    Bhuiyan, Didarul B; Middleton, John C; Tannenbaum, Rina; Wick, Timothy M

    2016-08-01

    A bone graft is a complicated structure that provides mechanical support and biological signals that regulate bone growth, reconstruction, and repair. A single-component material is inadequate to provide a suitable combination of structural support and biological stimuli to promote bone regeneration. Multicomponent composite biomaterials lack adequate bonding among the components to prevent phase separation after implantation. We have previously developed a novel multistep polymerization and fabrication process to construct a nano-hydroxyapatite-poly(D,L-lactide-co-glycolide)-collagen biomaterial (abbreviated nHAP-PLGA-collagen) with the components covalently bonded to each other. In the present study, the mechanical properties and osteogenic potential of nHAP-PLGA-collagen are characterized to assess the material's suitability to support bone regeneration. nHAP-PLGA-collagen films exhibit tensile strength very close to that of human cancellous bone. Human mesenchymal stem cells (hMSCs) are viable on 2D nHAP-PLGA-collagen films with a sevenfold increase in cell population after 7 days of culture. Over 5 weeks of culture, hMSCs deposit matrix and mineral consistent with osteogenic differentiation and bone formation. As a result of matrix deposition, nHAP-PLGA-collagen films cultured with hMSCs exhibit 48% higher tensile strength and fivefold higher moduli compared to nHAP-PLGA-collagen films without cells. More interestingly, secretion of matrix and minerals by differentiated hMSCs cultured on the nHAP-PLGA-collagen films for 5 weeks mitigates the loss of mechanical strength that accompanies PLGA hydrolysis. PMID:27120980

  16. Electrospun polyurethane/hydroxyapatite bioactive scaffolds for bone tissue engineering: the role of solvent and hydroxyapatite particles.

    PubMed

    Tetteh, G; Khan, A S; Delaine-Smith, R M; Reilly, G C; Rehman, I U

    2014-11-01

    Polyurethane (PU) is a promising polymer to support bone-matrix producing cells due to its durability and mechanical resistance. In this study two types of medical grade poly-ether urethanes Z3A1 and Z9A1 and PU-Hydroxyapatite (PU-HA) composites were investigated for their ability to act as a scaffold for tissue engineered bone. PU dissolved in varying concentrations of dimethylformamide (DMF) and tetrahydrofuran (THF) solvents were electrospun to attain scaffolds with randomly orientated non-woven fibres. Bioactive polymeric composite scaffolds were created using 15 wt% Z3A1 in a 70/30 DMF/THF PU solution and incorporating micro- or nano-sized HA particles in a ratio of 3:1 respectively, whilst a 25 wt% Z9A1 PU solution was doped in ratio of 5:1. Chemical properties of the resulting composites were evaluated by FTIR and physical properties by SEM. Tensile mechanical testing was carried out on all electrospun scaffolds. MLO-A5 osteoblastic mouse cells and human embryonic mesenchymal progenitor cells, hES-MPs were seeded on the scaffolds to test their biocompatibility and ability to support mineralised matrix production over a 28 day culture period. Cell viability was assayed by MTT and calcium and collagen deposition by Sirius red and alizarin red respectively. SEM images of both electrospun PU scaffolds and PU-HA composite scaffolds showed differences in fibre morphology with changes in solvent combinations and size of HA particles. Inclusion of THF eliminated the presence of beads in fibres that were present in scaffolds fabricated with 100% DMF solvent, and resulted in fibres with a more uniform morphology and thicker diameters. Mechanical testing demonstrated that the Young׳s Modulus and yield strength was lower at higher THF concentrations. Inclusion of both sizes of HA particles in PU-HA solutions reinforced the scaffolds leading to higher mechanical properties, whilst FTIR characterisation confirmed the presence of HA in all composite scaffolds. Although

  17. Hydroxyapatite grafted carbon nanotubes and graphene nanosheets: Promising bone implant materials

    NASA Astrophysics Data System (ADS)

    Oyefusi, Adebola; Olanipekun, Opeyemi; Neelgund, Gururaj M.; Peterson, Deforest; Stone, Julia M.; Williams, Ebonee; Carson, Laura; Regisford, Gloria; Oki, Aderemi

    2014-11-01

    In the present study, hydroxyapatite (HA) was successfully grafted to carboxylated carbon nanotubes (CNTs) and graphene nanosheets. The HA grafted CNTs and HA-graphene nanosheets were characterized using FT-IR, TGA, SEM and X-ray diffraction. The HA grafted CNTs and graphene nanosheets (CNTs-HA and Gr-HA) were further used to examine the proliferation and differentiation rate of temperature-sensitive human fetal osteoblastic cell line (hFOB 1.19). Total protein assays and western blot analysis of osteocalcin expression were used as indicators of cell proliferation and differentiation. Results indicated that hFOB 1.19 cells proliferate and differentiate well in treatment media containing CNTs-HA and graphene-HA. Both CNTs-HA and graphene-HA could be promising nanomaterials for use as scaffolds in bone tissue engineering.

  18. Accurate Fabrication of Hydroxyapatite Bone Models with Porous Scaffold Structures by Using Stereolithography

    NASA Astrophysics Data System (ADS)

    Maeda, Chiaki; Tasaki, Satoko; Kirihara, Soshu

    2011-05-01

    Computer graphic models of bioscaffolds with four-coordinate lattice structures of solid rods in artificial bones were designed by using a computer aided design. The scaffold models composed of acryl resin with hydroxyapatite particles at 45vol. % were fabricated by using stereolithography of a computer aided manufacturing. After dewaxing and sintering heat treatment processes, the ceramics scaffold models with four-coordinate lattices and fine hydroxyapatite microstructures were obtained successfully. By using a computer aided analysis, it was found that bio-fluids could flow extensively inside the sintered scaffolds. This result shows that the lattice structures will realize appropriate bio-fluid circulations and promote regenerations of new bones.

  19. Synthesis, characterization and osteoconductivity properties of bone fillers based on alendronate-loaded poly(ε-caprolactone)/hydroxyapatite microspheres.

    PubMed

    Chen, Jianhong; Luo, Yun; Hong, Liangqing; Ling, You; Pang, Jun; Fang, Youqiang; Wei, Kun; Gao, Xin

    2011-03-01

    A superior drug controlled release system capable of achieving efficient osteogenesis is in imperative demand because of limited bone substitute tissue for the treatment of bone defect. In the present study, we investigated the potential of using poly(ε-caprolactone)-hydroxyapatite (PCL-HA) composite microspheres as an injectable bone repair vehicle by controlled release of alendronate (AL), a medicine that belongs to the bisphosphonates family. The PCL/HA-AL microspheres were prepared with solid/oil/water emulsion technique, which included two processes: (1) AL was loaded on the hydroxyapatite nanoparticles; (2) the HA-AL complex was built in the PCL matrix. The spherical PCL/HA-AL microspheres were characterized with its significantly improved encapsulation efficiency of hydrophilic AL and better sustained release. Human bone mesenchymal stem cells (hMSCs) were cultured on the surface of these microspheres and exhibited high proliferative profile. Specifically, in osteogenic medium, hMSCs on the surface of PCL/HA-AL microspheres displayed superior osteogenic differentiation which was verified by alkaline phosphatase activity assay. In conclusion, by presenting strong osteogenic commitment of hMSCs in vitro, the PCL/HA-AL microspheres have the potential to be used as an injectable vehicle for local therapy of bone defect. PMID:21318627

  20. Design of polymer-biopolymer-hydroxyapatite biomaterials for bone tissue engineering: Through molecular control of interfaces

    NASA Astrophysics Data System (ADS)

    Verma, Devendra

    In this dissertation, novel biomaterials are designed for bone biomaterials and bone tissue engineering applications. Novel biomaterials of hydroxyapatite with synthetic and natural polymers have been fabricated using a combination of processing routes. Initially, we investigated hydroxyapatite-polycaprolactone-polyacrylic acid composites and observed that minimal interfacial interactions between polymer and mineral led to inadequate improvement in the mechanical properties. Bioactivity experiments on these composites showed that the presence of functional groups, such as carboxylate groups, influence bioactivity of the composites. We have developed and investigated composites of hydroxyapatite with chitosan and polygalacturonic acid (PgA). Chitosan and PgA are biocompatible, biodegradable, and also electrostatically complementary to each other. This strategy led to significant improvement in mechanical properties of new composites. The nanostructure analysis using atomic force microscopy revealed a multilevel organization in these composites. Enhancement in mechanical response was attributed to stronger interfaces due to strong electrostatic interaction between oppositely charged chitosan and PgA. Further analysis using the Rietveld method showed that biopolymers have marked impact on hydroxyapatite crystal growth and also on its crystal structure. Significant changes were observed in the lattice parameters of hydroxyapatite synthesized by following biomineralization method (organics mediated mineralization). For scaffold preparation, chitosan and PgA were mixed first, and then, nano-hydroxyapatite was added. Oppositely charged polyelectrolytes, such as chitosan and PgA, spontaneously form complex upon mixing. The poly-electrolyte complex exists as nano-sized particles. Chitosan/PgA scaffolds with and without hydroxyapatite were prepared by the freeze drying method. By controlling the rate of cooling and concentration, we have produced both fibrous and sheet

  1. Nanoparticles of cobalt-substituted hydroxyapatite in regeneration of mandibular osteoporotic bones

    PubMed Central

    Ignjatović, Nenad; Ajduković, Zorica; Savić, Vojin; Najman, Stevo; Mihailović, Dragan; Vasiljević, Perica; Stojanović, Zoran; Uskoković, Vuk; Uskoković, Dragan

    2012-01-01

    Indications exist that paramagnetic calcium phosphates may be able to promote regeneration of bone faster than their regular, diamagnetic counterparts. In this study, analyzed was the influence of paramagnetic cobalt-substituted hydroxyapatite nanoparticles on osteoporotic alveolar bone regeneration in rats. Simultaneously, biocompatibility of the material was tested in vitro, on osteoblastic MC3T3-E1 and epithelial Caco-2 cells in culture. The material was shown to be biocompatible and nontoxic when added to epithelial monolayers in vitro, while it caused a substantial decrease in the cell viability as well as deformation of the cytoskeleton and cell morphology when incubated with the osteoblastic cells. In the course of six months after the implantation of the material containing different amounts of cobalt, ranging from 5 – 12 wt%, in the osteoporotic alveolar bone of the lower jaw, the following parameters were investigated: histopathological parameters, alkaline phosphatase and alveolar bone density. The best result in terms of osteoporotic bone tissue regeneration was observed for hydroxyapatite nanoparticles with the largest content of cobalt ions. The histological analysis showed a high level of reparatory ability of the nanoparticulate material implanted in the bone defect, paralleled by a corresponding increase in the alveolar bone density. The combined effect of growth factors from autologous plasma admixed to cobalt-substituted hydroxyapatite was furthermore shown to have a crucial effect on the augmented osteoporotic bone regeneration upon the implantation of the biomaterial investigated in this study. PMID:23090835

  2. Analysis of human alveolar osteoblast behavior on a nano-hydroxyapatite substrate: an in vitro study

    PubMed Central

    2014-01-01

    Background Nano-hydroxyapatite (nHA) is a potential ideal biomaterial for bone regeneration. However, studies have yet to characterize the behavior of human osteoblasts derived from alveolar bone on nHA. Thus, the aim of the present study was to evaluate the influence of nHA on the adhesion, proliferation and differentiation of these alveolar bone-derived cells. Methods Primary human alveolar osteoblasts were collected from the alveolar ridge of a male periodontal patient during osseous resective surgery and grown on culture plates coated with either polylysine or polylysine with nano-hydroxyapatite (POL/nHA) composite. The cells were grown and observed for 14 days, and then assessed for potential modifications to osteoblasts homeostasis as evaluated by quantitative reverse transcriptase-polymerase chain reaction (real time RT-PCR), scanning electron microscopy and atomic force microscopy. Results Real time PCR revealed a significant increase in the expression of the selected markers of osteoblast differentiation (bone morphogenetic protein (BMP)-2,-5,-7, ALP, COLL-1A2, OC, ON) in cells grown on the POL/nHA substrate. In addition, as compared with the POL surface, cells grown on the POL/nHA substrate demonstrated better osteoconductive properties, as demonstrated by the increase in adhesion and spreading, likely as a result of the increased surface roughness of the composite. Conclusions The increased expression of BMPs and osteoinductive biomarkers suggest that nano-hydroxyapatite may stimulate the proliferation and differentiation of local alveolar osteoblasts and thus encourage bone regeneration at sites of alveolar bone regeneration. PMID:24650194

  3. Development of graded hydroxyapatite/CaCO(3) composite structures for bone ingrowth.

    PubMed

    Heilmann, F; Standard, O C; Müller, F A; Hoffman, M

    2007-09-01

    Ceramic composites composed of constituents with different bone cell reactions present an interesting consideration for a new bone replacement material. The first component of the composite used in this study, hydroxyapatite, is known to be replaced by natural tissue significantly slower than the second, calcium carbonate, which has limited structural stability. A graded hydroxyapatite/calcium carbonate composite with bimodal component distribution was developed using a combined slip infiltration and dip-coating technique from a porous polyurethane sponge replica. A graded hydroxyapatite scaffold with porosities from 5 to 90% was produced and then infiltrated with a calcium carbonate slip and sintered. The resultant composite had improved mechanical properties compared with the monolith as measured by crushing and moduli tests. PMID:17483903

  4. New Bismuth-Substituted Hydroxyapatite Nanoparticles for Bone Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Ciobanu, Gabriela; Bargan, Ana Maria; Luca, Constantin

    2015-11-01

    New bismuth-substituted hydroxyapatite [Ca10- x Bi x (PO4)6(OH)2 where x = 0-2.5] nanoparticles were synthesized by the co-precipitation method from aqueous solutions. The structural properties of the samples were analyzed by scanning electron microscopy coupled with x-ray analysis, x-ray powder diffraction, x-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and Brunauer-Emmett-Teller surface area analysis. The results confirm that bismuth ions have been incorporated into the hydroxyapatite lattice. The prepared nanocrystalline powders consisted of hydroxyapatite as single phase with hexagonal structure, crystal sizes smaller than 60 nm and (Bi + Ca)/P atomic ratio of around 1.67. The hydroxyapatite samples doped with Bi have mesoporous textures with pores size of around 2 nm and specific surface area in the range of 12-25 m2/g. The Bi-substituted hydroxyapatite powders are more effective against Gram-negative Escherichia coli bacteria than Gram-positive Staphylococcus aureus bacteria.

  5. Magnesium-enriched hydroxyapatite as bone filler in an ameloblastoma mandibular defect

    PubMed Central

    Grigolato, Roberto; Pizzi, Natalia; Brotto, Maria C; Corrocher, Giovanni; Desando, Giovanna; Grigolo, Brunella

    2015-01-01

    The aim of this study was to evaluate the clinical performance of a magnesium-enriched hydroxyapatite biomaterial used as bone substitute in a case of mandibular ameloblastoma treated with conservative surgery. A 63 year old male patient was treated for an ameloblastoma in the anterior mandibular profile. After tissue excision, the bone defect was filled with a synthetic hydroxyapatite biomaterial enriched with magnesium ions, in order to promote bone tissue regeneration and obtain a good aesthetic result. Twenty-five months after surgery, due to ameloblastoma recurrence in an area adjacent to the previously treated one, the patient underwent to a further surgery. In that occasion the surgeon performed a biopsy in the initially treated area, in order to investigate the nature of the newly-formed tissue and to evaluate the bone regenerative potential of this biomaterial by clinical, radiographic and histological analyses. The clinical, radiographic and histological evaluations showed various characteristics of bone remodeling stage with an ongoing osteogenic formation and a good osteo-integration. In conclusion, magnesium-enriched hydroxyapatite used as bone substitute in a mandibular defect due to ameloblastoma excision showed an effective bone regeneration at 25 months follow-up, demonstrating an excellent biocompatibility and a high osteo-integration property. PMID:25784998

  6. [Structural features of ectopic bone-like tissue in porous hydroxyapatite blocks].

    PubMed

    Hattori, Shigetaka

    2008-06-01

    Calcium phosphate ceramics are widely used as bone substitutes because of their favorable biocompatibility and osteoconductivity. Some studies have reported that porous hydroxyapatite (HA), with its special structure, implanted at ectopic (non-bone) sites, exhibits osteoinductive activity. However, few studies have examined the detailed ultrastructure of mineralized tissue formed in biomaterials like HA. Therefore, it is important to examine whether the mineralized tissue exhibits structural characteristics of normal bone tissue. Thus, in this study, we subcutaneously implanted hydroxyapatite blocks (5 x 5 x 7 mm) into the backs of adult dogs, and performed detailed morphological examination of the bone-like tissue formed, which showed a lamellar structure. Immunohistochemical staining revealed that the matrix fibers of the bone-like tissue were mainly composed of type I collagen. The pattern of crystal deposition in matrix fibers and the structure of osteoblast-, osteocyte-, bone-lining-, and osteoclast-like cells were similar to those of normal bone tissue and osteogenic cells, respectively. These results indicate that cellular events observed in normal bone formation and remodeling occur in bone-like tissue as well, and suggest that bone-like tissue in HA blocks is very similar to bone tissue. PMID:18634458

  7. Development of a synthetic bone scaffold using porous hydroxyapatite for bone repair.

    PubMed

    Mustaffa, R; Besar, I; Andanastuti, M

    2008-07-01

    In this study, porous hydroxyapatite (HA) samples were fabricated via sponge techniques with the aid of sago as part of the binder mixture. Development processes for the production of porous bone graft substitutes are studied using polyurethane sponge. To obtain the optimum amount of binder for successful fabrication of porous HA were done. Initially, porous HA powder was synthesized using calcium hydroxide and orthorphosphoric acid. Meanwhile, sago was mixed with PVA in a certain ratio to be used as binder for preparing the porous HA. After a series of investigative tests were conducted to characterize the sintered samples, the use of the sago and polymeric mixture was found to successfully aid the fabrication of porous HA samples. In this investigation, comparison of physical and mechanical characteristics between samples prepared using difference techniques was made. PMID:19025001

  8. Enhanced bioactivity of a poly(propylene fumarate) bone graft substitute by augmentation with nano-hydroxyapatite.

    PubMed

    Lewandrowski, Kai-Uwe; Bondre, Shrikar P; Wise, Donald L; Trantolo, Debra J

    2003-01-01

    The bioactivity of a nano-hydroxyapatite-augmented, bioresorbable bone graft substitute made from the unsaturated polyester, poly(propylene fumarate), was analyzed by evaluating biocompatibility and osteointegration of implants placed into a rat tibial defect. Three groups of eight animals each were evaluated by grouting bone graft substitutes into 3-mm holes that were made into the anteromedial tibial metaphysis of rats. Thus, a total of 24 animals was included in this study. Two different formulations varying as to the type of hydroxyapatite were used: Group 1 - nano-hydroxyapatite, Group 2 - micron-hydroxyapatite, with a Group 3 control defect remaining unfilled. Animals of each of the three groups were sacrificed in groups of eight at postoperative week three. Histologic analysis revealed best superior biocompatibility and osteointegration of bone graft substitutes when nanohydroxyapatite was employed. At three weeks, there was more reactive new bone formation in this group when compared to the micron-hydroxyapatite group. The control group showed incomplete closure of the defect. This study suggested that nano-hydroxyapatite may improve upon the bioactivity of bone implant and repair materials. The model scaffold used in this study, poly(propylene fumarate), appeared to provide an osteoconductive pathway by which bone will grow in faster. Clinical implications of the use potential advantages of nano-hydroxyapatite on bone repair and orthopaedic implant design are discussed. PMID:12775902

  9. Development of model hydroxyapatite bone scaffolds with multiscale porosity for potential load bearing applications

    NASA Astrophysics Data System (ADS)

    Dellinger, Jennifer Gwynne

    2005-11-01

    Model hydroxyapatite (HA) bone scaffolds consisting of a latticed pattern of rods were fabricated by a solid freeform fabrication (SFF) technique based on the robotic deposition of colloidal pastes. An optimal HA paste formulation for this method was developed. Local porosity, i.e. microporosity (1--30 mum) and sintering porosity (less than 1 mum), were produced by including polymer microsphere porogens in the HA pastes and by controlling the sintering of the scaffolds. Scaffolds with and without local porosity were evaluated with and without in vitro accelerated degradation. Percent weight loss of the scaffolds and calcium and phosphorus concentrations in solution increased with degradation time. After degradation, compressive strength and modulus decreased significantly for scaffolds with local porosity, but did not change significantly for scaffolds without local porosity. The compressive strength and modulus of scaffolds without local porosity were comparable to human cortical bone and were significantly greater than the scaffolds with local porosity. Micropores in HA disks caused surface pits that increased the surface roughness as compared to non-microporous HA disks. Mouse mesenchymal stem cells extended their cell processes into these microporous pits on HA disks in vitro. ALP expression was prolonged, cell attachment strength increased, and ECM production appeared greater on microporous HA disks compared to non-microporous HA disks and tissue culture treated polystyrene controls. Scaffolds with and without microporosity were implanted in goats bones. Microporous scaffolds with rhBMP-2 increased the percent of the scaffold filled with bone tissue compared to microporous scaffolds without rhBMP-2. Lamellar bone inside scaffolds was aligned near the rods junctions whereas lamellar bone was aligned in a more random configuration away from the rod junctions. Microporous scaffolds stained darkly with toluidine blue beneath areas of contact with new bone. This

  10. Oleic acid surfactant in polycaprolactone/hydroxyapatite-composites for bone tissue engineering.

    PubMed

    Cardoso, Guinea B C; Maniglio, Devid; Volpato, Fabio Z; Tondon, Abhishek; Migliaresi, Claudio; Kaunas, Roland R; Zavaglia, Cecilia A C

    2016-08-01

    Bone substitutes are required to repair osseous defects caused by a number of factors, such as traumas, degenerative diseases, and cancer. Autologous bone grafting is typically used to bridge bone defects, but suffers from chronic pain at the donor-site and limited availability of graft material. Tissue engineering approaches are being investigated as viable alternatives, which ideal scaffold should be biocompatible, biodegradable, and promote cellular interactions and tissue development, need to present proper mechanical and physical properties. In this study, poly(ε-caprolactone) (PCL), oleic acid (OA) and hydroxyapatite (HAp) were used to obtain films whose properties were investigated by contact angle, scanning electron microscopy, atomic force microscopy, tensile mechanical tests, and in vitro tests with U2OS human osteosarcoma cells by direct contact. Our results indicate that by using OA as surfactant/dispersant, it was possible to obtain a homogenous film with HAp. The PCL/OA/Hap sample had twice the roughness of the control (PCL) and a lower contact angle, indicating increased hydrophilicity of the film. Furthermore, mechanical testing showed that the addition of HAp decreased the load at yield point and tensile strength and increased tensile modulus, indicating a more brittle composition vs. PCL matrix. Preliminary cell culture experiments carried out with the films demonstrated that U2OS cells adhered and proliferated on all surfaces. The data demonstrate the improved dispersion of HAp using OA and the important consequences of this addition on the composite, unveiling the potentially of this composition for bone growth support. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1076-1082, 2016. PMID:26033969

  11. Nano-hydroxyapatite/chitosan-starch nanocomposite as a novel bone construct: Synthesis and in vitro studies.

    PubMed

    Shakir, Mohammad; Jolly, Reshma; Khan, Mohd Shoeb; Iram, Noor e; Khan, Haris M

    2015-09-01

    A novel ternary nanocomposite system incorporating hydroxyapatite, chitosan and starch (n-HA/CS-ST) has been synthesized by co-precipitation method at room temperature, addressing the issues of biocompatibility, mechanical strength and cytotoxicity required for bone tissue engineering. The interactions, crystallite size, surface morphology and thermal stability against n-HA/CS nanocomposite have been obtained by comparing the results of FTIR, SEM, TEM, DLS, XRD and TGA/DTA. A comparative study of bioactivity and thermal stability of n-HA/CS and n-HA/CS-ST nanocomposites revealed that the incorporation of starch as templating agent enhanced these properties in n-HA/CS-ST nanocomposite. A lower swelling rate of n-HA/CS-ST relative to n-HA/CS indicates a higher mechanical strength supportive of bone tissue ingrowths. The MTT assay on murine fibroblast L929 and human osteoblasts-like MG-63 cells and in vitro bioactivity of n-HA/CS-ST matrix referred superior non-toxic nature of n-HA/CS-ST nanocomposite and greater possibility of osteointegration in vivo respectively. Furthermore n-HA/CS-ST exhibited improved antibacterial property against both Gram-positive and Gram-negative bacteria relative to n-HA/CS. PMID:26116779

  12. A new approach to enhancement of bone formation by electrically polarized hydroxyapatite.

    PubMed

    Teng, N C; Nakamura, S; Takagi, Y; Yamashita, Y; Ohgaki, M; Yamashita, K

    2001-10-01

    An electrical field may affect osteogenesis. Since we found that hydroxyapatite (HA) ceramics may be polarizable, we hypothesized that electrically polarized HA may foster production of new bone in vivo. Both polarized and non-polarized HA ceramics were inserted into the subperiosteum spaces at the parietal bone area of rats. After 2, 4, and 8 weeks, the implant sites were examined histologically. Morphometric analysis revealed that new bone formation was accelerated on the negatively charged surface of the polarized HA (N-surface) at 2 weeks. The newly formed bone approached maturation at 4 weeks and was thicker on the N-surface than in the controls. By 8 weeks, newly formed bone in the controls was almost the same as that on the N-surface. These findings suggest that polarized HA is biocompatible and that bone formation on the N-surface is enhanced in the early stage of bone healing. PMID:11706953

  13. Effect of loading time on marginal bone loss around hydroxyapatite-coated implants

    PubMed Central

    Kim, Young-Kyun; Ahn, Kyo-Jin; Yun, Pil-Young; Kim, Minkyoung; Yang, Hong-So; Yi, Yang-Jin

    2013-01-01

    Objectives The objective of this study is compare the rate of marginal bone resorption around hydroxyapatite-coated implants given different loading times in order to evaluate their stability. Materials and Methods The study was conducted retrospectively for one year, targeting 41 patients whose treatment areas were the posterior maxilla and the mandible. Osstem TS III HA (Osstem Implant Co., Busan, Korea) and Zimmer TSV-HA (Zimmer Dental, Carlsbad, CA, USA), which employ the new hydroxyapatite coating technique, were used. The patients were divided into two groups - immediate and delayed loading - and the bone level at the time of loading commencement and after one year of loading was measured using periapical radiography. Differences between the groups were evaluated using Mann-Whitney (α=0.05). Results For all patients as a single group, the survival rate of the implants was 100%, and the mean marginal bone loss was 0.26±0.59 mm. In comparison of the differences by loading, mean marginal bone loss of 0.32±0.69 mm was recorded for the immediate loading group whereas the delayed loading group had mean marginal bone loss of 0.16±0.42 mm. However, the difference was not significant (P>0.05). Conclusion Within the limited observation period of one year, predictable survival rates can be expected when using immediately loaded hydroxyapatite-coated implants. PMID:24471037

  14. Enhancement of Periprosthetic Bone Quality with Topical Hydroxyapatite-Bisphosphonate Composite

    PubMed Central

    Suratwala, Sanjeev J.; Cho, Samuel K.; van Raalte, Jonathan J.; Park, Sang Hyun; Seo, Sung Wook; Chang, Seong-Sil; Gardner, Thomas R.; Lee, Francis Young-In

    2008-01-01

    Background: Implant loosening is associated with inflammatory bone loss induced by ultra-high molecular weight polyethylene wear debris. We hypothesized that a hydroxyapatite-bisphosphonate composite improves periprosthetic bone quality and osseous integration of an intramedullary implant even in the presence of ultra-high molecular weight polyethylene particles in an experimental rat femur model. Methods: A preliminary in vitro study determined the optimal concentration of zoledronate (50 μM) that would maximally decrease osteoclasts without harming osteoblasts. Hydroxyapatite-coated intramedullary nails were implanted bilaterally in the femora of sixteen rats (the control group), and hydroxyapatite-zoledronate-coated nails were implanted bilaterally in the femora of sixteen rats (the experimental group). Ultra-high molecular weight polyethylene particles were introduced into the femoral canal before implantation. Eight rats from each group were killed at six weeks, and the remaining rats were killed at six months. Periprosthetic bone mass was analyzed by dual x-ray absorptiometry and microcomputed tomography. Osseous integration was examined by biomechanical testing of pullout strength. Results: The mean bone area (and standard deviation) in the periprosthetic bone region was significantly greater (p < 0.0001) in the hydroxyapatite-zoledronate group (2.388 ± 0.960 mm2) than in the control group (0.933 ± 0.571 mm2). This difference was larger in the six-week group than in the six-month group (p = 0.03). The average peak pullout force for the treated femora (241.0 ± 95.1 N) was significantly greater (p < 0.0001) than that for the controls (55.6 ± 49.0 N). This difference was similar in the six-week and six-month groups. The energy required for nail pullout was significantly greater (p < 0.0001) for the treated femora (521.6 ± 293.8 N-mm) than for the controls (142.2 ± 152.1 N-mm). This difference in energy to pullout was similar in the six-week and six

  15. Synthetical bone-like and biological hydroxyapatites: a comparative study of crystal structure and morphology.

    PubMed

    Marković, Smilja; Veselinović, Ljiljana; Lukić, Miodrag J; Karanović, Ljiljana; Bračko, Ines; Ignjatović, Nenad; Uskoković, Dragan

    2011-08-01

    Phase composition, crystal structure and morphology of biological hydroxyapatite (BHAp) extracted from human mandible bone, and carbonated hydroxyapatite (CHAp), synthesized by the chemical precipitation method, were studied by x-ray powder diffraction (XRD), Fourier transform infrared (FTIR) and Raman (R) spectroscopy techniques, combined with transmission electron microscopy (TEM). Structural and microstructural parameters were determined through Rietveld refinement of recorded XRD data, performed using the FullProf computing program, and TEM. Microstructural analysis shows anisotropic extension along the [00l] crystallographic direction (i.e. elongated crystallites shape) of both investigated samples. The average crystallite sizes of 10 and 8 nm were estimated for BHAp and CHAp, respectively. The FTIR and R spectroscopy studies show that carbonate ions substitute both phosphate and hydroxyl ions in the crystal structure of BHAp as well as in CHAp, indicating that both of them are mixed AB-type of CHAp. The thermal behaviour and carbonate content were analysed using thermogravimetric and differential thermal analysis. The carbonate content of about 1 wt.% and phase transition, at near 790 °C, from HAp to β-tricalcium phosphate were determined in both samples. The quality of synthesized CHAp powder, particularly, the particle size distribution and uniformity of morphology, was analysed by a particle size analyser based on laser diffraction and field emission scanning electron microscopy, respectively. These data were used to discuss similarity between natural and synthetic CHAp. Good correlation between the unit cell parameters, average crystallite size, morphology, carbonate content and crystallographic positions of carbonate ions in natural and synthetic HAp samples was found. PMID:21659698

  16. Development of high strength hydroxyapatite for bone tissue regeneration using nanobioactive glass composites

    SciTech Connect

    Shrivastava, Pragya; Dalai, Sridhar; Vijayalakshmi, S.; Sudera, Prerna; Sivam, Santosh Param; Sharma, Pratibha

    2013-02-05

    With an increasing demand of biocompatible bone substitutes for the treatment of bone diseases and bone tissue regeneration, bioactive glass composites are being tested to improvise the osteoconductive as well as osteoinductive properties. Nanobioactive glass (nBG) composites, having composition of SiO{sub 2} 70 mol%, CaO 26 mol % and P{sub 2}O{sub 5} 4 mol% were prepared by Freeze drying method using PEG-PPG-PEG co-polymer. Polymer addition improves the mechanical strength and porosity of the scaffold of nBG. Nano Bioactive glass composites upon implantation undergo specific reactions leading to the formation of crystalline hydroxyapatite (HA). This is tested in vitro using Simulated Body Fluid (SBF). This high strength hydroxyapatite (HA) layer acts as osteoconductive in cellular environment, by acting as mineral base of bones, onto which new bone cells proliferate leading to new bone formation. Strength of the nBG composites as well as HA is in the range of cortical and cancellous bone, thus proving significant for bone tissue regeneration substitutes.

  17. Hydroxyapatite-calcium sulfate-hyaluronic acid composite encapsulated with collagenase as bone substitute for alveolar bone regeneration.

    PubMed

    Subramaniam, Sadhasivam; Fang, Yen-Hsin; Sivasubramanian, Savitha; Lin, Feng-Huei; Lin, Chun-pin

    2016-01-01

    Periodontitis is a very severe inflammatory condition of the periodontium that progressively damages the soft tissue and destroys the alveolar bone that supports the teeth. The bone loss is naturally irreversible because of limited reparability of the teeth. Advancement in tissue engineering provides an effective regeneration of osseous defects with suitable dental implants or tissue-engineered constructs. This study reports a hydroxyapatite, calcium sulfate hemihydrate and hyaluronic acid laden collagenase (HAP/CS/HA-Col) as a bone substitute for the alveolar bone regeneration. The composite material was mechanically tested and the biocompatibility was evaluated by WST-1 assay. The in vivo bone formation was assessed in rat with alveolar bone defects and the bone augmentation by the HAP/CS/HA-Col composite was confirmed by micro-CT images and histological examination. The mechanical strength of 6.69 MPa with excellent biocompatibility was obtained for the HAP/CS/HA-Col composite. The collagenase release profile had facilitated the acceleration of bone remodeling process and it was confirmed by the findings of micro-CT and H&E staining. The bone defects implanted with HAP/CS/HA composite containing 2 mg/mL type I collagenase have shown improved new bone formation with matured bone morphology in comparison with the HAP/CS/HA composite that lacks the collagenase and the porous hydroxyapatite (p-HAP) granules. The said findings demonstrated that the collagenase inclusion in HAP/CS/HA composite is a feasible approach for the alveolar bone regeneration and the same design can also be applied to other defective tissues. PMID:26454048

  18. Templated repair of long bone defects in rats with bioactive spiral-wrapped electrospun amphiphilic polymer/hydroxyapatite scaffolds.

    PubMed

    Kutikov, Artem B; Skelly, Jordan D; Ayers, David C; Song, Jie

    2015-03-01

    Effective repair of critical-size long bone defects presents a significant clinical challenge. Electrospun scaffolds can be exploited to deliver protein therapeutics and progenitor cells, but their standalone application for long bone repair has not been explored. We have previously shown that electrospun composites of amphiphilic poly(d,l-lactic acid)-co-poly(ethylene glycol)-co-poly(d,l-lactic acid) (PELA) and hydroxyapatite (HA) guide the osteogenic differentiation of bone marrow stromal cells (MSCs), making these scaffolds uniquely suited for evaluating cell-based bone regeneration approaches. Here we examine whether the in vitro bioactivity of these electrospun scaffolds can be exploited for long bone defect repair, either through the participation of exogenous MSCs or through the activation of endogenous cells by a low dose of recombinant human bone morphogenetic protein-2 (rhBMP-2). In critical-size rat femoral segmental defects, spiral-wrapped electrospun HA-PELA with preseeded MSCs resulted in laminated endochondral ossification templated by the scaffold across the longitudinal span of the defect. Using GFP labeling, we confirmed that the exogenous MSCs adhered to HA-PELA survived at least 7 days postimplantation, suggesting direct participation of these exogenous cells in templated bone formation. When loaded with 500 ng of rhBMP-2, HA-PELA spirals led to more robust but less clearly templated bone formation than MSC-bearing scaffolds. Both treatment groups resulted in new bone bridging over the majority of the defect by 12 weeks. This study is the first demonstration of a standalone bioactive electrospun scaffold for templated bone formation in critical-size long bone defects. PMID:25695310

  19. Collagen-Hydroxyapatite Scaffolds Induce Human Adipose Derived Stem Cells Osteogenic Differentiation In Vitro.

    PubMed

    Calabrese, Giovanna; Giuffrida, Raffaella; Fabbi, Claudia; Figallo, Elisa; Lo Furno, Debora; Gulino, Rosario; Colarossi, Cristina; Fullone, Francesco; Giuffrida, Rosario; Parenti, Rosalba; Memeo, Lorenzo; Forte, Stefano

    2016-01-01

    Mesenchymal stem cells (MSCs) play a crucial role in regulating normal skeletal homeostasis and, in case of injury, in bone healing and reestablishment of skeletal integrity. Recent scientific literature is focused on the development of bone regeneration models where MSCs are combined with biomimetic three-dimensional scaffolds able to direct MSC osteogenesis. In this work the osteogenic potential of human MSCs isolated from adipose tissue (hADSCs) has been evaluated in vitro in combination with collagen/Mg doped hydroxyapatite scaffolds. Results demonstrate the high osteogenic potential of hADSCs when cultured in specific differentiation induction medium, as revealed by the Alizarin Red S staining and gene expression profile analysis. In combination with collagen/hydroxyapatite scaffold, hADSCs differentiate into mature osteoblasts even in the absence of specific inducing factors; nevertheless, the supplement of the factors markedly accelerates the osteogenic process, as confirmed by the expression of specific markers of pre-osteoblast and mature osteoblast stages, such as osterix, osteopontin (also known as bone sialoprotein I), osteocalcin and specific markers of extracellular matrix maturation and mineralization stages, such as ALPL and osteonectin. Hence, the present work demonstrates that the scaffold per se is able to induce hADSCs differentiation, while the addition of osteo-inductive factors produces a significant acceleration of the osteogenic process. This observation makes the use of our model potentially interesting in the field of regenerative medicine for the treatment of bone defects. PMID:26982592

  20. Collagen-Hydroxyapatite Scaffolds Induce Human Adipose Derived Stem Cells Osteogenic Differentiation In Vitro

    PubMed Central

    Fabbi, Claudia; Figallo, Elisa; Lo Furno, Debora; Gulino, Rosario; Colarossi, Cristina; Fullone, Francesco; Giuffrida, Rosario; Parenti, Rosalba; Memeo, Lorenzo; Forte, Stefano

    2016-01-01

    Mesenchymal stem cells (MSCs) play a crucial role in regulating normal skeletal homeostasis and, in case of injury, in bone healing and reestablishment of skeletal integrity. Recent scientific literature is focused on the development of bone regeneration models where MSCs are combined with biomimetic three-dimensional scaffolds able to direct MSC osteogenesis. In this work the osteogenic potential of human MSCs isolated from adipose tissue (hADSCs) has been evaluated in vitro in combination with collagen/Mg doped hydroxyapatite scaffolds. Results demonstrate the high osteogenic potential of hADSCs when cultured in specific differentiation induction medium, as revealed by the Alizarin Red S staining and gene expression profile analysis. In combination with collagen/hydroxyapatite scaffold, hADSCs differentiate into mature osteoblasts even in the absence of specific inducing factors; nevertheless, the supplement of the factors markedly accelerates the osteogenic process, as confirmed by the expression of specific markers of pre-osteoblast and mature osteoblast stages, such as osterix, osteopontin (also known as bone sialoprotein I), osteocalcin and specific markers of extracellular matrix maturation and mineralization stages, such as ALPL and osteonectin. Hence, the present work demonstrates that the scaffold per se is able to induce hADSCs differentiation, while the addition of osteo-inductive factors produces a significant acceleration of the osteogenic process. This observation makes the use of our model potentially interesting in the field of regenerative medicine for the treatment of bone defects. PMID:26982592

  1. Enhanced bone healing using collagen-hydroxyapatite scaffold implantation in the treatment of a large multiloculated mandibular aneurysmal bone cyst in a thoroughbred filly.

    PubMed

    David, Florent; Levingstone, Tanya J; Schneeweiss, Wilfried; de Swarte, Marie; Jahns, Hanne; Gleeson, John P; O'Brien, Fergal J

    2015-10-01

    An unmet need remains for a bone graft substitute material that is biocompatible, biodegradable and capable of promoting osteogenesis safely in vivo. The aim of this study was to investigate the use of a novel collagen-hydroxyapatite (CHA) bone graft substitute in the clinical treatment of a mandibular bone cyst in a young horse and to assess its potential to enhance repair of the affected bone. A 2 year-old thoroughbred filly, presenting with a multilobulated aneurysmal bone cyst, was treated using the CHA scaffold. Post-operative clinical follow-up was carried out at 2 weeks and 3, 6 and 14 months. Cortical thickening in the affected area was observed from computed tomography (CT) examination as early as 3 months post-surgery. At 14 months, reduced enlargement of the operated mandible was observed, with no fluid-filled area. The expansile cavity was occupied by moderately dense mineralized tissue and fat and the compact bone was remodelled, with a clearer definition between cortex and medulla observed. This report demonstrates the promotion of enhanced bone repair following application of the CHA scaffold material in this craniomaxillofacial indication, and thus the potential of this material for translation to human applications. PMID:25712436

  2. Effect of sintered silicate-substituted hydroxyapatite on remodelling processes at the bone-implant interface.

    PubMed

    Porter, Alexandra E; Patel, Nelesh; Skepper, Jeremy N; Best, Serena M; Bonfield, William

    2004-07-01

    Phase pure, sintered granules of hydroxyapatite (HA) and silicon-substituted hydroxyapatite (Si-HA) were implanted for 6 and 12 weeks in an ovine model. Samples containing the bone-implant interface were prepared for ultramicrotomy and transmission electron microscopy (TEM) using an anhydrous sample preparation procedure. The results demonstrate that the morphology of apatite deposits and the sequence of events at the interfaces of bone with pure HA and with Si-HA implants, were different. Organised collagen fibrils were first found at the bone/Si-HA interface after 6 weeks, whereas they were found only after 12 weeks around the pure HA implant. Many more nodular aggregates comprised of plate-like apatite crystallites were observed in the vicinity of Si-HA than around the pure HA after 12 weeks in vivo. These findings suggest that the incorporation of silicate ions into HA promotes processes of bone remodelling at the bone/HA interface. TEM observations suggested that the trabecular bone weaves over the Si-HA and that the collagen fibrils form a mechanical interlock with the Si-HA ceramic implants. High-resolution lattice imaging illustrated apatite crystallites contiguous with the Si-HA ceramic and revealed a direct relationship between the bone mineral and the Si-HA ceramic. PMID:14980425

  3. Dissolution of synthetic and bovine bone-derived hydroxyapatites fabricated by hot-pressing

    NASA Astrophysics Data System (ADS)

    Kim, Young Gook; Seo, Dong Seok; Lee, Jong Kook

    2008-11-01

    Dissolving behavior of hydroxyapatite (HA) ceramics prepared from bovine bone (BHA) was investigated and compared with synthetic HA. BHA power was obtained by calcining at 800 °C for 1 h to remove organics and then attritor-milling for 24 h. Synthetic HA and BHA powders were prepared by hot-pressing at 1000 °C for 0.5 h under the pressure of 30 MPa in Ar atmosphere. Sintered densities of the synthetic HA and the BHA were about 96% and 95%, respectively. The BHA consisted of mainly HA and small amount of magnesium oxide. Immersion test revealed that there was no clear evidence of dissolution for the BHA, whereas extensive dissolution on the surface of the synthetic HA occurred in buffered water. Accordingly, dense hydroxyapatite from bovine bone was more stable than synthetic HA in liquid environment.

  4. Biomimetic scaffolds based on hydroxyapatite nanorod/poly(D,L) lactic acid with their corresponding apatite-forming capability and biocompatibility for bone-tissue engineering.

    PubMed

    Nga, Nguyen Kim; Hoai, Tran Thanh; Viet, Pham Hung

    2015-04-01

    This study presents a facile synthesis of biomimetic hydroxyapatite nanorod/poly(D,L) lactic acid (HAp/PDLLA) scaffolds with the use of solvent casting combined with a salt-leaching technique for bone-tissue engineering. Field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive X-ray spectroscopy were used to observe the morphologies, pore structures of synthesized scaffolds, interactions between hydroxyapatite nanorods and poly(D,L) lactic acid, as well as the compositions of the scaffolds, respectively. Porosity of the scaffolds was determined using the liquid substitution method. Moreover, the apatite-forming capability of the scaffolds was evaluated through simulated body fluid (SBF) incubation tests, whereas the viability, attachment, and distribution of human osteoblast cells (MG 63 cell line) on the scaffolds were determined through alamarBlue assay and confocal laser microscopy after nuclear staining with 4',6-diamidino-2-phenylindole and actin filaments of a cytoskeleton with Oregon Green 488 phalloidin. Results showed that hydroxyapatite nanorod/poly(D,L) lactic acid scaffolds that mimic the structure of natural bone were successfully produced. These scaffolds possessed macropore networks with high porosity (80-84%) and mean pore sizes ranging 117-183 μm. These scaffolds demonstrated excellent apatite-forming capabilities. The rapid formation of bone-like apatites with flower-like morphology was observed after 7 days of incubation in SBFs. The scaffolds that had a high percentage (30 wt.%) of hydroxyapatite demonstrated better cell adhesion, proliferation, and distribution than those with low percentages of hydroxyapatite as the days of culture increased. This work presented an efficient route for developing biomimetic composite scaffolds, which have potential applications in bone-tissue engineering. PMID:25791418

  5. Specific Biomimetic Hydroxyapatite Nanotopographies Enhance Osteoblastic Differentiation and Bone Graft Osteointegration

    PubMed Central

    Loiselle, Alayna E.; Wei, Lai; Faryad, Muhammad; Paul, Emmanuel M.; Lewis, Gregory S.; Gao, Jun; Lakhtakia, Akhlesh

    2013-01-01

    Impaired healing of cortical bone grafts represents a significant clinical problem. Cadaveric bone grafts undergo extensive chemical processing to decrease the risk of disease transmission; however, these processing techniques alter the bone surface and decrease the osteogenic potential of cells at the healing site. Extensive work has been done to optimize the surface of bone grafts, and hydroxyapatite (HAP) and nanotopography both increase osteoblastic differentiation. HAP is the main mineral component of bone and can enhance osteoblastic differentiation and bone implant healing in vivo, while nanotopography can enhance osteoblastic differentiation, adhesion, and proliferation. This is the first study to test the combined effects of HAP and nanotopographies on bone graft healing. With the goal of identifying the optimized surface features to improve bone graft healing, we tested the hypothesis that HAP-based nanotopographic resurfacing of bone grafts improves integration of cortical bone grafts by enhancing osteoblastic differentiation. Here we show that osteoblastic cells cultured on processed bones coated with specific-scale (50–60 nm) HAP nanotopographies display increased osteoblastic differentiation compared to cells on uncoated bone, bones coated with poly-l-lactic acid nanotopographies, or other HAP nanotopographies. Further, bone grafts coated with 50–60-nm HAP exhibited increased formation of new bone and improved healing, with mechanical properties equivalent to live autografts. These data indicate the potential for specific HAP nanotopographies to not only increase osteoblastic differentiation but also improve bone graft incorporation, which could significantly increase patient quality of life after traumatic bone injuries or resection of an osteosarcoma. PMID:23510012

  6. Nano-hydroxyapatite composite biomaterials for bone tissue engineering--a review.

    PubMed

    Venkatesan, Jayachandran; Kim, Se-Kwon

    2014-10-01

    In recent years, significant development has been achieved in the construction of artificial bone with ceramics, polymers and metals. Nano-hydroxyapatite (nHA) is widely used bioceramic material for bone graft substitute owing to its biocompatibility and osteoconductive properties. nHA with chitin, chitosan, collagen, gelatin, fibrin, polylactic acid, polycaprolactone, poly(lactic-co-glycolic) acid, polyamide, polyvinyl alcohol, polyurethane and polyhydroxybutyrate based composite scaffolds have been explored in the present review for bone graft substitute. This article further reviews the preparative methods, chemical interaction, biocompatibiity, biodegradation, alkaline phosphatase activity, mineralization effect, mechanical properties and delivery of nHA-based nanocomposites for bone tissue regeneration. The nHA based composite biomaterials proved to be promising biomaterials for bone tissue engineering. PMID:25992432

  7. Hydroxyapatite-magnetite-MWCNT nanocomposite as a biocompatible multifunctional drug delivery system for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Pistone, Alessandro; Iannazzo, Daniela; Panseri, Silvia; Montesi, Monica; Tampieri, Anna; Galvagno, Signorino

    2014-10-01

    New magnetic hydroxyapatite-based nanomaterials as bone-specific systems for controlled drug delivery have been synthesized. The synthesized hydroxyapatite, HA, decorated with magnetite nanoparticles by a deposition method (HA/Fe3O4) and the nanocomposite system obtained using magnetic multi-walled carbon nanotubes (HA/MWCNT/Fe3O4) as a filler for HA have been characterized by chemical and morphological analyses, and their biological behavior was investigated. The systems have also been doped with clodronate in order to combine the effect of bone biomineralization induced by hydroxyapatite-based composites with the decrease of osteoclast formation induced by the drug. An analysis of the preosteoclastic RAW264.7 cell proliferation by MTT assay confirmed the high biocompatibility of the three systems. TRAP staining of RAW 264.7 conditioned with sRAKL to induce osteoclastogenesis, cultured in the presence of the systems doped and undoped with clodronate, showed the inhibitory effect of clodronate after we counted the MNC TRAP+cells but only in the osteoclast formation; in particular, the system HA/Fe3O4-Clo exerted a high inhibitory effect compared to the drug alone. These results demonstrate that the synthesized nanocomposites are a biocompatible magnetic drug delivery system and can represent a useful multimodal platform for applications in bone tissue engineering.

  8. Electrospun Hydroxyapatite-Containing Chitosan Nanofibers Crosslinked with Genipin for Bone Tissue Engineering

    PubMed Central

    Frohbergh, Michael E.; Katsman, Anna; Botta, Gregory P.; Lazarovici, Phillip; Schauer, Caroline L.; Wegst, Ulrike G. K.; Lelkes, Peter I.

    2012-01-01

    Reconstruction of large bone defects remains problematic in orthopedic and craniofacial clinical practice. Autografts are limited in supply and are associated with donor site morbidity while other materials show poor integration with the host’s own bone. This lack of integration is often due to the absence of periosteum, the outer layer of bone that contains osteoprogenitor cells and is critical for the growth and remodeling of bone tissue. In this study we developed a one-step platform to electrospin nanofibrous scaffolds from chitosan, which also contain hydroxyapatite nanoparticles and are crosslinked with genipin. We hypothesized that the resulting composite scaffolds represent a microenvironment that emulates the physical, mineralized structure and mechanical properties of non-weight bearing bone extracellular matrix while promoting osteoblast differentiation and maturation similar to the periosteum. The ultrastructure and physicochemical properties of the scaffolds were studied using scanning electron microscopy and spectroscopic techniques. The average fiber diameters of the electrospun scaffolds were 227±154 nm as spun, and increased to 335±119 nm after crosslinking with genipin. Analysis by X-ray diffraction, Fourier transformed infrared spectroscopy and energy dispersive spectroscopy confirmed the presence of characteristic features of hydroxyapatite in the composite chitosan fibers. The Young’s modulus of the composite fibrous scaffolds was 142±13 MPa, which is similar to that of the natural periosteum. Both pure chitosan scaffolds and composite hydroxyapatite-containing chitosan scaffolds supported adhesion, proliferation and osteogenic differentiation of mouse 7F2 osteoblast-like cells. Expression and enzymatic activity of alkaline phosphatase, an early osteogenic marker, were higher in cells cultured on the composite scaffolds as compared to pure chitosan scaffolds, reaching a significant, 2.4 fold, difference by day 14 (p<0.05). Similarly

  9. Novel cerium doped glass-reinforced hydroxyapatite with antibacterial and osteoconductive properties for bone tissue regeneration.

    PubMed

    Morais, D S; Fernandes, S; Gomes, P S; Fernandes, M H; Sampaio, P; Ferraz, M P; Santos, J D; Lopes, M A; Sooraj Hussain, N

    2015-09-01

    The aim of this work was to develop a bioactive bone substitute with an effective antibacterial ability based on a cerium (Ce) doped glass-reinforced hydroxyapatite (GR-HA) composite. Developed composites were physicochemically characterized, using x-ray diffraction (XRD) analysis, SEM, energy dispersive x-ray spectroscopy (EDS), and flexural bending strength (FBS) tests. X-ray photoelectron spectroscopy (XPS) analysis was performed to analyze the oxidation state of Ce in the prepared doped glass. The antimicrobial activity of the composites was evaluated against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa; whether the cytocompatibility profile was assayed with human osteoblastic-like cells (Mg-63 cell line). The results revealed that the Ce inclusion in the GR-HA matrix induced the antimicrobial ability of the composite. In addition, Ce-doped materials reported an adequate biological behavior following seeding of osteoblastic populations, by inducing cell adhesion and proliferation. Developed materials were also found to enhance the expression of osteoblastic-related genes. Overall, the developed GR-HA_Ce composite is a prospective candidate to be used within the clinical scenario with a successful performance due to the effective antibacterial properties and capability of enhancing the osteoblastic cell response. PMID:26391473

  10. Combined effect of strontium and zoledronate on hydroxyapatite structure and bone cell responses.

    PubMed

    Boanini, Elisa; Torricelli, Paola; Gazzano, Massimo; Della Bella, Elena; Fini, Milena; Bigi, Adriana

    2014-07-01

    The influence of the simultaneous presence of the two inhibitors of bone degradation, strontium and zoledronate, on the direct synthesis of hydroxyapatite was explored in the range of Sr concentration up to 50 atom% at two different bisphosphonate concentrations (ZOL7 and ZOL14). The results of structural analysis indicated that HA can be obtained as a single crystalline phase up to a Sr concentration in solution of 20 and 10 atom% within the ZOL7 and ZOL14 series respectively. Both Sr substitution and ZOL incorporation affect the length of the coherently scattering crystalline domains and the dimensions of HA nanocrystals. At greater Sr content, XRD full profile fitting data indicate that zoledronate provokes the segregation of Sr in two crystalline apatitic phases, at different strontium content. Co-cultures of osteoblast-like MG63 cells and human osteoclast show that ZOL displays a greater inhibitory influence than Sr on osteoclast proliferation and activity. On the other hand, the results obtained on osteoblast surnatant and on gene expression indicate that Sr exerts a greater promotion on osteoblast proliferation and differentiation. The co-presence of Sr and ZOL has a combined effect on the differentiation markers, so that HA containing about 4 wt% ZOL and 4 Sr atom%, and even more HA containing about 4 wt% ZOL and 8 Sr atom%, result the best compromise for osteoblast promotion and osteoclast inhibition. PMID:24731709

  11. Primary human osteoblast culture on 3D porous collagen-hydroxyapatite scaffolds.

    PubMed

    Jones, Gemma L; Walton, Robin; Czernuszka, Jan; Griffiths, Sarah L; El Haj, Alicia J; Cartmell, Sarah H

    2010-09-15

    There is a need in tissue-engineering for 3D scaffolds that mimic the natural extracellular matrix of bone to enhance cell adhesion, proliferation, and differentiation. The scaffold is also required to be degradable. A highly porous scaffold has been developed to incorporate two of the extracellular components found in bone-collagen and hydroxyapatite (HA). The scaffold's collagen component is an afibrillar monomeric type I atelocollagen extracted from foetal calf's skin. This provided a novel environment for the inclusion of HA powder. Five hundred thousand primary human osteoblasts were seeded onto 4 mm cubed scaffolds that varied in ratio of HA to collagen. Weight ratios of 1:99, 25:75, 50:50, and 75:25 hydroxyapatite:collagen (HA:Collagen) were analysed. The scaffolds plus cells were cultured for 21 days. DNA assays and live/dead viability staining demonstrated that all of the scaffolds supported cell proliferation and viability. An alkaline phosphatase assay showed similar osteoblast phenotype maintenance on all of the 3D scaffolds analysed at 21 days. MicroCT analysis demonstrated an increase in total sample volume (correlating to increase in unmineralised matrix production). An even distribution of HA throughout the collagen matrix was observed using this technique. Also at 3 weeks, reductions in the percentage of the mineralised phase of the constructs were seen. These results indicate that each of the ratios of HA/collagen scaffolds have great potential for bone tissue engineering. PMID:20694991

  12. Development of gelatin-chitosan-hydroxyapatite based bioactive bone scaffold with controlled pore size and mechanical strength.

    PubMed

    Maji, Kanchan; Dasgupta, Sudip; Kundu, Biswanath; Bissoyi, Akalabya

    2015-01-01

    Hydroxyapatite-chitosan/gelatin (HA:Chi:Gel) nanocomposite scaffold has potential to serve as a template matrix to regenerate extra cellular matrix of human bone. Scaffolds with varying composition of hydroxyapatite, chitosan, and gelatin were prepared using lyophilization technique where glutaraldehyde (GTA) acted as a cross-linking agent for biopolymers. First, phase pure hydroxyapatite-chitosan nanocrystals were in situ synthesized by coprecipitation method using a solution of 2% acetic acid dissolved chitosan and aqueous solution of calcium nitrate tetrahydrate [Ca(NO3)2,4H2O] and diammonium hydrogen phosphate [(NH4)2H PO4]. Keeping solid loading constant at 30 wt% and changing the composition of the original slurry of gelatin, HA-chitosan allowed control of the pore size, its distribution, and mechanical properties of the scaffolds. Microstructural investigation by scanning electron microscopy revealed the formation of a well interconnected porous scaffold with a pore size in the range of 35-150 μm. The HA granules were uniformly dispersed in the gelatin-chitosan network. An optimal composition in terms of pore size and mechanical properties was obtained from the scaffold with an HA:Chi:Gel ratio of 21:49:30. The composite scaffold having 70% porosity with pore size distribution of 35-150 μm exhibited a compressive strength of 3.3-3.5 MPa, which is within the range of that exhibited by cancellous bone. The bioactivity of the scaffold was evaluated after conducting mesenchymal stem cell (MSC) - materials interaction and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay using MSCs. The scaffold found to be conducive to MSC's adhesion as evident from lamellipodia, filopodia extensions from cell cytoskeleton, proliferation, and differentiation up to 14 days of cell culture. PMID:26335156

  13. In Vivo Bone Formation Within Engineered Hydroxyapatite Scaffolds in a Sheep Model.

    PubMed

    Lovati, A B; Lopa, S; Recordati, C; Talò, G; Turrisi, C; Bottagisio, M; Losa, M; Scanziani, E; Moretti, M

    2016-08-01

    Large bone defects still represent a major burden in orthopedics, requiring bone-graft implantation to promote the bone repair. Along with autografts that currently represent the gold standard for complicated fracture repair, the bone tissue engineering offers a promising alternative strategy combining bone-graft substitutes with osteoprogenitor cells able to support the bone tissue ingrowth within the implant. Hence, the optimization of cell loading and distribution within osteoconductive scaffolds is mandatory to support a successful bone formation within the scaffold pores. With this purpose, we engineered constructs by seeding and culturing autologous, osteodifferentiated bone marrow mesenchymal stem cells within hydroxyapatite (HA)-based grafts by means of a perfusion bioreactor to enhance the in vivo implant-bone osseointegration in an ovine model. Specifically, we compared the engineered constructs in two different anatomical bone sites, tibia, and femur, compared with cell-free or static cell-loaded scaffolds. After 2 and 4 months, the bone formation and the scaffold osseointegration were assessed by micro-CT and histological analyses. The results demonstrated the capability of the acellular HA-based grafts to determine an implant-bone osseointegration similar to that of statically or dynamically cultured grafts. Our study demonstrated that the tibia is characterized by a lower bone repair capability compared to femur, in which the contribution of transplanted cells is not crucial to enhance the bone-implant osseointegration. Indeed, only in tibia, the dynamic cell-loaded implants performed slightly better than the cell-free or static cell-loaded grafts, indicating that this is a valid approach to sustain the bone deposition and osseointegration in disadvantaged anatomical sites. PMID:27075029

  14. In vivo study on hydroxyapatite scaffolds with trabecular architecture for bone repair.

    PubMed

    Appleford, Mark R; Oh, Sunho; Oh, Namsik; Ong, Joo L

    2009-06-15

    The objective of this research was to investigate the bone formation and angio-conductive potential of hydroxyapatite (HA) scaffolds closely matched to trabecular bone in a canine segmental defect after 3 and 12 weeks post implantation. Histomorphometric comparisons were made between naturally forming trabecular bone (control) and defects implanted with scaffolds fabricated with micro-size (M-HA) and nano-size HA (N-HA) ceramic surfaces. Scaffold architecture was similar to trabecular bone formed in control defects at 3 weeks. No significant differences were identified between the two HA scaffolds; however, significant bone in-growth was observed by 12 weeks with 43.9 +/- 4.1% and 50.4 +/- 8.8% of the cross-sectional area filled with mineralized bone in M-HA and N-HA scaffolds, respectively. Partially organized, lamellar collagen fibrils were identified by birefringence under cross-polarized light at both 3 and 12 weeks post implantation. Substantial blood vessel infiltration was identified in the scaffolds and compared with the distribution and diameter of vessels in the surrounding cortical bone. Vessels were less numerous but significantly larger than native cortical Haversian and Volkmann canals reflecting the scaffold architecture where open spaces allowed interconnected channels of bone to form. This study demonstrated the potential of trabecular bone modeled, highly porous and interconnected, HA scaffolds for regenerative orthopedics. PMID:18478555

  15. Preparation and Functional Assessment of Composite Chitosan-Nano-Hydroxyapatite Scaffolds for Bone Regeneration

    PubMed Central

    Reves, Benjamin T.; Jennings, Jessica A.; Bumgardner, Joel D.; Haggard, Warren O.

    2012-01-01

    Composite chitosan-nano-hydroxyapatite microspheres and scaffolds prepared using a co-precipitation method have shown potential for use in bone regeneration. The goal of this research was to improve the functional properties of the composite scaffolds by modifying the fabrication parameters. The effects of degree of deacetylation (DDA), drying method, hydroxyapatite content and an acid wash on scaffold properties were investigated. Freeze-dried 61% DDA scaffolds degraded faster (3.5 ± 0.5% mass loss) than air-dried 61% DDA scaffolds and 80% DDA scaffolds, but had a lower compressive modulus of 0.12 ± 0.01 MPa. Air-dried 80% DDA scaffolds displayed the highest compressive modulus (3.79 ± 0.51 MPa) and these scaffolds were chosen as the best candidate for use in bone regeneration. Increasing the amount of hydroxyapatite in the air-dried 80% DDA scaffolds did not further increase the compressive modulus of the scaffolds. An acid wash procedure at pH 6.1 was found to increase the degradation of air-dried 80% DDA scaffolds from 1.3 ± 0.1% to 4.4 ± 0.4%. All of the formulations tested supported the proliferation of SAOS-2 cells. PMID:24956519

  16. Hydroxyapatite-hybridized chitosan/chitin whisker bionanocomposite fibers for bone tissue engineering applications.

    PubMed

    Pangon, Autchara; Saesoo, Somsak; Saengkrit, Nattika; Ruktanonchai, Uracha; Intasanta, Varol

    2016-06-25

    Biomimetic nanofibrous scaffolds derived from natural biopolymers for bone tissue engineering applications require good mechanical and biological performances including biomineralization. The present work proposes the utility of chitin whisker (CTWK) to enhance mechanical properties of chitosan/poly(vinyl alcohol) (CS/PVA) nanofibers and to offer osteoblast cell growth with hydroxyapatite (HA) mineralization. By using diacid as a solvent, electrospun CS/PVA nanofibrous membranes containing CTWK can be easily obtained. The dimension stability of nanofibrous CS/PVA/CTWK bionanocomposite is further controlled by exposing to glutaraldehyde vapor. The nanofibrous membranes obtained allow mineralization of HA in concentrated simulated body fluid resulting in an improvement of Young's modulus and tensile strength. The CTWK combined with HA in bionanocomposite is a key to promote osteoblast cell adhesion and proliferation. The present work, for the first time, demonstrates the use of CTWKs for bionanocomposite fibers of chitosan and its hydroxyapatite biomineralization with the function in osteoblast cell culture. These hydroxyapatite-hybridized CS/PVA/CTWK bionanocomposite fibers (CS/PVA/CTWK-HA) offer a great potential for bone tissue engineering applications. PMID:27083834

  17. High-solid-content hydroxyapatite slurry for the production of bone substitute scaffolds.

    PubMed

    Cunningham, E; Dunne, N; Walker, G; Buchanan, F

    2009-08-01

    Key to various bone substitute scaffold production techniques is the development of free-flowing ceramic slurry with optimum theological properties. The aim is to achieve a colloidal suspension with as high a solid content as possible while maintaining a low viscosity which easily penetrates the pores of relevant sacrificial templates. The following investigation describes the optimization of a hydroxyapatite slip and demonstrates its potential application in scaffold production. Using predominantly spherical particles of hydroxyapatite of between 0.82 microm and 16.2 microm, coupled with a 2 wt % addition of the anionic polyelectrolyte, ammonium polyacrylate, an 80 wt % (55.9 vol%) hydroxyapatite solid loaded slip with a viscosity of approximately 126mPas has been developed. Its ability to infiltrate and replicate porous preforms has been shown using polyurethane foam. The enhanced particle packing achieved has allowed for the production of scaffolds with highly dense and uniform grain structures. The results represent a significant improvement in current slurry production techniques and can be utilized to develop high-density ceramic bone substitute scaffolds. PMID:19743638

  18. Magnetic Hydroxyapatite Bone Substitutes to Enhance Tissue Regeneration: Evaluation In Vitro Using Osteoblast-Like Cells and In Vivo in a Bone Defect

    PubMed Central

    Panseri, Silvia; Cunha, Carla; D'Alessandro, Teresa; Sandri, Monica; Russo, Alessandro; Giavaresi, Gianluca; Marcacci, Maurilio; Hung, Clark T.; Tampieri, Anna

    2012-01-01

    In case of degenerative disease or lesion, bone tissue replacement and regeneration is an important clinical goal. In particular, nowadays, critical size defects rely on the engineering of scaffolds that are 3D structural supports, allowing cellular infiltration and subsequent integration with the native tissue. Several ceramic hydroxyapatite (HA) scaffolds with high porosity and good osteointegration have been developed in the past few decades but they have not solved completely the problems related to bone defects. In the present study we have developed a novel porous ceramic composite made of HA that incorporates magnetite at three different ratios: HA/Mgn 95/5, HA/Mgn 90/10 and HA/Mgn 50/50. The scaffolds, consolidated by sintering at high temperature in a controlled atmosphere, have been analysed in vitro using human osteoblast-like cells. Results indicate high biocompatibility, similar to a commercially available HA bone graft, with no negative effects arising from the presence of magnetite or by the use of a static magnetic field. HA/Mgn 90/10 was shown to enhance cell proliferation at the early stage. Moreover, it has been implanted in vivo in a critical size lesion of the rabbit condyle and a good level of histocompatibility was observed. Such results identify this scaffold as particularly relevant for bone tissue regeneration and open new perspectives for the application of a magnetic field in a clinical setting of bone replacement, either for magnetic scaffold fixation or magnetic drug delivery. PMID:22685602

  19. Direct Scaffolding of Biomimetic Hydroxyapatite-gelatin Nanocomposites using Aminosilane Cross-linker for Bone Regeneration

    PubMed Central

    Chiu, Chi-Kai; Ferreira, Joao; Luo, Tzy-Jiun M.; Geng, Haixia; Lin, Feng-Chang; Ko, Ching-Chang

    2012-01-01

    Hydroxyapatite-gelatin modified siloxane (GEMOSIL) nanocomposite was developed by coating, kneading and hardening processes to provide formable scaffolding for alloplastic graft applications. The present study aims to characterize scaffolding formability and mechanical properties of GEMOSIL, and to test the in vitro and in vivo biocompatibility of GEMOSIL. Buffer Solution initiated formable paste followed by the sol-gel reaction led to a final hardened composite. Results showed the adequate coating of aminosilane, 11–19 wt%, affected the cohesiveness of the powders and the final compressive strength (69 MPa) of the composite. TGA and TEM results showed the effective aminosilane coating that preserves hydroxyapatite-gelatin nanocrystals from damage. Both GEMOSIL with and without titania increased the mineralization of preosteoblasts in vitro. Only did titania additives revealed good in vivo bone formation in rat calvarium defects. The scaffolding formability, due to cohesive bonding among GEMOSIL particles, could be further refined to fulfill the complicated scaffold processes. PMID:22669282

  20. Hydroxyapatite nanocrystals functionalized with alendronate as bioactive components for bone implant coatings to decrease osteoclastic activity

    NASA Astrophysics Data System (ADS)

    Bosco, Ruggero; Iafisco, Michele; Tampieri, Anna; Jansen, John A.; Leeuwenburgh, Sander C. G.; van den Beucken, Jeroen J. J. P.

    2015-02-01

    The integration of bone implants within native bone tissue depends on periprosthetic bone quality, which is severely decreased in osteoporotic patients. In this work, we have synthesized bone-like hydroxyapatite nanocrystals (nHA) using an acid-base neutralization reaction and analysed their physicochemical properties. Subsequently, we have functionalized the nHA with alendronate (nHAALE), a well-known bisphosphonate drug used for the treatment of osteoporosis. An in vitro osteoclastogenesis test was carried out to evaluate the effect of nHAALE on the formation of osteoclast-like cells from monocytic precursor cells (i.e. RAW264.7 cell line) showing that nHAALE significantly promoted apoptosis of osteoclast-like cells. Subsequently, nHA and nHAALE were deposited on titanium disks using electrospray deposition (ESD), for which characterisation of the deposited coatings confirmed the presence of alendronate in nHAALE coatings with nanoscale thickness of about 700 nm. These results indicate that alendronate linked to hydroxyapatite nanocrystals has therapeutic potential and nHAALE can be considered as an appealing coating constituent material for orthopaedic and oral implants for application in osteoporotic patients.

  1. Diode λ830nm laser associated with hydroxyapatite and biological membranes: bone repair in rats

    NASA Astrophysics Data System (ADS)

    Carneiro, Vanda S. M.; Limeira, Francisco d. A.; Gerbi, Marleny E. M.; Menezes, Rebeca F. d.; Santos-Neto, Alexandrino P. d.; Araújo, Natália C.

    2016-02-01

    The aim of the present study was to histologically assess the effect of laser therapy (AsGaAl, 830nm, 40mW, CW, φ ~0,6mm, 16J/cm2 per session, four points of 4J/cm2) on the repair of surgical defects created in the femur of Wistar rats. Background data: Several techniques have been proposed for the correction of bone defects, including the use of grafts and membranes. Despite the increase in the use of laser therapy for the biomodulation of bone repair, very few studies have assessed the associations between laser light and biomaterials. Method: The defects were filled with synthetic micro granular hydroxyapatite (HA) Gen-phos® implants and associated with bovine bone membranes (Gen-derm®). Surgical bone defects were created in 48 rats and divided into four groups: Group IA (control, n=12); Group IB (laser, n=12); Group IIA (HA + membrane, n=12); Group IIB (HA + membrane + laser, n=12). The irradiated groups received the first irradiation immediately after surgery. This radiation was then repeated seven times every 48h. The animals were sacrificed after 15, 21, and 30 days. Results: When comparing the groups irradiated with implants and membranes, it was found that the repair of the defects submitted to laser therapy occurred more quickly, starting 15 and 21 days after surgery. By the 30th day, the level of repair of the defects was similar in the irradiated and the non-irradiated groups. New bone formation was confirmed inside the cavity by the implant's osteoconduction. In the irradiated groups, there was an increment of this new bone formation. Conclusions: In conclusion, the use of laser therapy, particularly when associated with hydroxyapatite and biological membranes, produced a positive biomodulation effect on the healing process of bone defects on the femurs of rats.

  2. Porous nano-hydroxyapatite/collagen scaffold containing drug-loaded ADM-PLGA microspheres for bone cancer treatment.

    PubMed

    Rong, Zi-Jie; Yang, Lian-Jun; Cai, Bao-Ta; Zhu, Li-Xin; Cao, Yan-Lin; Wu, Guo-Feng; Zhang, Zan-Jie

    2016-05-01

    To develop adriamycin (ADM)-encapsulated poly(lactic-co-glycolic acid) (PLGA) nanoparticles in a porous nano-hydroxyapatite/collagen scaffold (ADM-PLGA-NHAC). To provide novel strategies for future treatment of osteosarcoma, the properties of the scaffold, including its in vitro extended-release properties, the inhibition effects of ADM-PLGA-NHAC on the osteosarcoma MG63 cells, and its bone repair capacity, were investigated in vivo and in vitro. The PLGA copolymer was utilized as a drug carrier to deliver ADM-PLGA nanoparticles (ADM-PLGA-NP). Porous nano-hydroxyapatite and collagen were used to materials to produce the porous nano-hydroxyapatite/collagen scaffold (NHAC), into which the ADM-PLGA-NP was loaded. The performance of the drug-carrying scaffold was assessed using multiple techniques, including scanning electron microscopy and in vitro extended release. The antineoplastic activities of scaffold extracts on the human osteosarcoma MG63 cell line were evaluated in vitro using the cell counting kit-8 (CCK8) method and live-dead cell staining. The bone repair ability of the scaffold was assessed based on the establishment of a femoral condyle defect model in rabbits. ADM-PLGA-NHAC and NHAC were implanted into the rat muscle bag for immune response experiments. A tumor-bearing nude mice model was created, and the TUNEL and HE staining results were observed under optical microscopy to evaluate the antineoplastic activity and toxic side effects of the scaffold. The composite scaffold demonstrated extraordinary extended-release properties, and its extracts also exhibited significant inhibition of the growth of osteosarcoma MG63 cells. In the bone repair experiment, no significant difference was observed between ADM-PLGA-NHAC and NHAC by itself. In the immune response experiments, ADM-PLGA-NHAC exhibited remarkable biocompatibility. The in vivo antitumor experiment revealed that the implantation of ADM-PLGA-NHAC in the tumor resulted in a improved antineoplastic

  3. Adaptive Bone Remodeling of the Femoral Bone After Tumor Resection Arthroplasty With an Uncemented Proximally Hydroxyapatite-Coated Stem.

    PubMed

    Andersen, Mikkel R; Petersen, Michael M

    2016-01-01

    Loss of bone stock and stress shielding is a significant challenge in limb salvage surgery. This study investigates the adaptive bone remodeling of the femoral bone after implantation of a tumor prosthesis with an uncemented press fit stem. We performed a prospective 1 yr follow-up of 6 patients (mean age: 55 (26-78) yr, female/male=3/3) who underwent bone tumor resection surgery of the proximal femur (n=3) or distal femur (n=3). Reconstruction was done using a Global Modular Replacement System (Stryker® Orthopaedics, Mahwah, NJ) tumor prosthesis, and all patients received a straight-fluted 125-mm uncemented press-fit titanium alloy stem with hydroxyapatite coating of the proximal part of the stem. Measurements of bone mineral density (BMD; g/cm2) were done postoperatively and after 3, 6, and 12 mo in the part of the femur bone containing the Global Modular Replacement System stem using dual-energy X-ray absorptiometry. BMD was measured in 3 regions of interest (ROIs) in the femur bone. Nonparametric analysis of variance (Friedman test) for evaluation of changes in BMD over time. BMD decreased in all 3 ROIs with time. In ROI 1 (p=0.01), BMD decreased by 10% after 3 mo and ended with a total decrease of 14% after 1 yr. In ROI 2 (p=0.006), BMD was decreased by 6% after 3 and 6 mo; after 1 yr of follow-up, BMD was 9% below the postoperative value. In ROI 3 (p=0.009), BMD decreased by 6% after 3 and 6 mo; after 1 yr of follow-up, BMD was 8% below the postoperative value. A bone loss of 8%-9% during the first postoperative year was seen along the femoral stem, but in the bone containing the hydroxyapatite-coated part of the stem, the decrease in BMD was 14%, thus indicating that stress shielding of this part of the bone may play a role for the adaptive bone remodeling. PMID:25843447

  4. Fabrication of porous hydroxyapatite scaffolds as artificial bone preform and its biocompatibility evaluation.

    PubMed

    Jang, Dong-Woo; Franco, Rose Ann; Sarkar, Swapan Kumar; Lee, Byong-Taek

    2014-01-01

    In this study, a novel porous hydroxyapatite scaffold was designed and fabricated to imitate natural bone through a multipass extrusion process. The conceptual design manifested unidirectional microchannels at the exterior part of the scaffold to facilitate rapid biomineralization and a central canal that houses the bone marrow. External and internal fissures were minimized during microwave sintering at 1,100 °C. No deformation was noted, and a mechanically stable scaffold was fabricated. Detailed microstructure of the fabricated artificial bone was examined by scanning electron microscope and X-ray diffractometer, and material properties like compressive strength were evaluated. The initial biocompatibility was examined by the cell proliferation of MG-63 osteoblast-like cells using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Preliminary in vivo investigation in a rabbit model after 4 weeks and 8 weeks of implantation showed full osteointegration of the scaffold with the native tissue, and formation of bone tissue within the pore network, as examined by microcomputed tomography analyses and histological staining. Osteon-like bone microarchitecture was observed along the unidirectional channel with microblood vessels. These confirm a biomimetic regeneration model in the implanted bone scaffold, which can be used as an artificial alternative for damaged bone. PMID:24399056

  5. Histological Comparison in Rats between Carbonate Apatite Fabricated from Gypsum and Sintered Hydroxyapatite on Bone Remodeling.

    PubMed

    Ayukawa, Yasunori; Suzuki, Yumiko; Tsuru, Kanji; Koyano, Kiyoshi; Ishikawa, Kunio

    2015-01-01

    Carbonate apatite (CO3Ap), the form of apatite found in bone, has recently attracted attention. The purpose of the present study was to histologically evaluate the tissue/cellular response toward the low-crystalline CO3Ap fabricated using a dissolution-precipitation reaction with set gypsum as a precursor. When set gypsum was immersed in a 100°C 1 mol/L Na3PO4 aqueous solution for 24 h, the set gypsum transformed into CO3Ap. Both CO3Ap and sintered hydroxyapatite (s-HAp), which was used as a control, were implanted into surgically created tibial bone defects of rats for histological evaluation. Two and 4 weeks after the implantation, histological sections were created and observed using light microscopy. The CO3Ap granules revealed both direct apposition of the bone matrix by osteoblasts and osteoclastic resorption. In contrast, the s-HAp granules maintained their contour even after 4 weeks following implantation which implied that there was a lack of replacement into the bone. The s-HAp granules were sometimes encapsulated with fibrous tissue, and macrophage polykaryon was occasionally observed directly apposed to the implanted granules. From the viewpoint of bone remodeling, the CO3Ap granules mimicked the bone matrix, suggesting that CO3Ap may be an appropriate bone substitute. PMID:26504813

  6. A mechanical and histomorphometric analysis of bone bonding by hydroxyapatite-coated strain gages.

    PubMed

    Wilson, D L; Szivek, J A; Anderson, P L; Miera, V L; Battraw, G A

    1998-01-01

    Identification of the strains controlling bone remodeling is important for determining ways to prevent bone loss due to load deprivation, or implant placement. Long-term monitoring of strains can potentially provide the best information. Glues are resorbed within 2-3 weeks. Two formulations of microcrystalline hydroxyapatite (HA) were used to attach strain gages to rat femora to assess their long-term in vivo strain measurement capability. Seven male rats received HA-coated gages, and 2 animals underwent a sham procedure. The gages were prepared using a published technique and placed on the antero-lateral aspect of the left femora. After 6-7 weeks, the animals were euthanized and both femora explanted. Gages were attached to the right femora with cyanoacrylate. All femora were tested in cantilever bending, then embedded, sectioned, and stained with mineralized bone stain. The undecalcified sections were examined using transmitted and ultraviolet light microscopy. Mechanical testing showed one HA formulation provided 70-100% bonding. Histology showed intimate contact between the gage and bone surface. Histomorphometry indicated increased bone activity under the gage compared to the remaining bone, the controls, and the shams. The results indicate that microcrystalline HAs bond to bone quickly and can allow long term in vivo measurements. PMID:9659635

  7. Vacuum plasma sprayed coatings using ionic silver doped hydroxyapatite powder to prevent bacterial infection of bone implants.

    PubMed

    Guimond-Lischer, Stefanie; Ren, Qun; Braissant, Olivier; Gruner, Philipp; Wampfler, Bruno; Maniura-Weber, Katharina

    2016-06-01

    Fast and efficient osseointegration of implants into bone is of crucial importance for their clinical success; a process that can be enhanced by coating the implant surface with hydroxyapatite (HA) using the vacuum plasma spray technology (VPS). However, bacterial infections, especially the biofilm formation on implant surfaces after a surgery, represent a serious complication. With ever-increasing numbers of antibiotic-resistant bacteria, there is great interest in silver (Ag) as an alternative to classical antibiotics due to its broad activity against Gram-positive and Gram-negative bacterial strains. In the present study, silver ions were introduced into HA spray powder by ion exchange and the HA-Ag powder was applied onto titanium samples by VPS. The Ag-containing surfaces were evaluated for the kinetics of the silver release, its antibacterial effect against Staphylococcus aureus as well as Escherichia coli, and possible cytotoxicity against human bone cells. The HA-Ag coatings with different concentrations of Ag displayed mechanical and compositional properties that fulfill the regulatory requirements. Evaluation of the Ag release kinetic showed a high release rate in the first 24 h followed by a decreasing release rate over the four subsequent days. The HA-Ag coatings showed no cytotoxicity to primary human bone cells while exhibiting antibacterial activity to E. coli and S. aureus. PMID:26964530

  8. Effects of strontium ions on growth and dissolution of hydroxyapatite and on bone mineral detection.

    PubMed

    Christoffersen, J; Christoffersen, M R; Kolthoff, N; Bärenholdt, O

    1997-01-01

    Preparation and analyses of a series of hydroxyapatites (HA) containing 1-10 mol % of Ca2+ replaced by Sr2+ is reported. The solubility of these apatites is found to increase with increasing content of Sr2+, 10% SrHA dissolves faster than CaHA at given values of Ca2+ and phosphate concentrations, but with a similar rate at the same degree of saturation. Sr2+ is found to inhibit the rates of both dissolution and growth of CaHA and 10% SrHA at pH 7.2, CaHA being more strongly inhibited by Sr2+ than 10% SrHA. The effect of partial substitution of Ca2+ in hydroxyapatite by Sr2+ on bone mineral content (BMC) and bone mineral density (BMD) measured by dual energy X-ray absorptiometry has been studied using three commercial densitometers. Extrapolating the absorption data for up to 10% replacement of Ca2+ by Sr2+ to 100% substitution of Ca2+ by Sr2+ in HA leads to an apparent increase in BMC or BMD of about a factor of 10. This factor is in agreement with theoretical calculations using attenuation coefficients of the atoms concerned. It is concluded that existing BMC scanners register artificially high values of BMC if the bone contains significant amounts of Sr2+ or other metal ions with atomic number larger than calcium. PMID:8988347

  9. Polyether ether ketone implants achieve increased bone fusion when coated with nano-sized hydroxyapatite: a histomorphometric study in rabbit bone

    PubMed Central

    Johansson, Pär; Jimbo, Ryo; Naito, Yoshihito; Kjellin, Per; Currie, Fredrik; Wennerberg, Ann

    2016-01-01

    Polyether ether ketone (PEEK) possesses excellent mechanical properties similar to those of human bone and is considered the best alternative material other than titanium for orthopedic spine and trauma implants. However, the deficient osteogenic properties and the bioinertness of PEEK limit its fields of application. The aim of this study was to limit these drawbacks by coating the surface of PEEK with nano-scaled hydroxyapatite (HA) minerals. In the study, the biological response to PEEK, with and without HA coating, was investigated. Twenty-four screw-like and apically perforated implants in the rabbit femur were histologically evaluated at 3 weeks and 12 weeks after surgery. Twelve of the 24 implants were HA coated (test), and the remaining 12 served as uncoated PEEK controls. At 3 weeks and 12 weeks, the mean bone–implant contact was higher for test compared to control (P<0.05). The bone area inside the threads was comparable in the two groups, but the perforating hole showed more bone area for the HA-coated implants at both healing points (P<0.01). With these results, we conclude that nano-sized HA coating on PEEK implants significantly improved the osteogenic properties, and in a clinical situation this material composition may serve as an implant where a rapid bone fusion is essential. PMID:27103801

  10. Biodegradation rate of shellac coated bovine hydroxyapatite for bone filler material

    NASA Astrophysics Data System (ADS)

    Triyono, Joko; Triyono, Susilowati, Endang; Murdiyantara, Suci Anindya

    2016-03-01

    This work reports on the effect of shellac coated hydroxyapatite (HA) on the biodegradation rate. The HA was processed from bovine bone. Shellac was derived from the resinous secretion of the lac insect. The aims of the addition of shellac solution is to know how the biodegradation rate material in the Phosphate Buffered Saline (PBS) solution. The four different of shellac solutions (2,5%; 5%; 7,5%; and 10% weight) coated HA scaffoldand one ratio as a control. It was concluded that the ability of biodegradation rate a materialwas not influenced by the ratio of shellac. All materials were biodegradedwhen they were soaked in PBS solution.

  11. Self-assembled right handed helical ribbons of the bone mineral hydroxyapatite.

    PubMed

    Suganthi, R V; Girija, E K; Narayana Kalkura, S; Varma, H K; Rajaram, A

    2009-12-01

    A self-assembled right handed helical ribbon of bone mineral, hydroxyapatite (HAp) was crystallized in sodium meta silicate gel matrix at 27 degrees C and the physiological pH (7.4). At temperatures 37 and 47 degrees C, helical structures were followed by many Liesegang rings. The samples were characterized by FT-IR, XRD, SEM, ICP-OES and TG-DTA techniques. The helical ribbon consisted of platy Ca-deficient apatite crystals of size 2.8 microm. Liesegang ring had a continuous network of fibers with interconnected pores. The samples exhibited bioactivity when soaked in SBF. PMID:18568388

  12. Magnetic poly(ε-caprolactone)/iron-doped hydroxyapatite nanocomposite substrates for advanced bone tissue engineering

    PubMed Central

    Gloria, A.; Russo, T.; D'Amora, U.; Zeppetelli, S.; D'Alessandro, T.; Sandri, M.; Bañobre-López, M.; Piñeiro-Redondo, Y.; Uhlarz, M.; Tampieri, A.; Rivas, J.; Herrmannsdörfer, T.; Dediu, V. A.; Ambrosio, L.; De Santis, R.

    2013-01-01

    In biomedicine, magnetic nanoparticles provide some attractive possibilities because they possess peculiar physical properties that permit their use in a wide range of applications. The concept of magnetic guidance basically spans from drug delivery and hyperthermia treatment of tumours, to tissue engineering, such as magneto-mechanical stimulation/activation of cell constructs and mechanosensitive ion channels, magnetic cell-seeding procedures, and controlled cell proliferation and differentiation. Accordingly, the aim of this study was to develop fully biodegradable and magnetic nanocomposite substrates for bone tissue engineering by embedding iron-doped hydroxyapatite (FeHA) nanoparticles in a poly(ε-caprolactone) (PCL) matrix. X-ray diffraction analyses enabled the demonstration that the phase composition and crystallinity of the magnetic FeHA were not affected by the process used to develop the nanocomposite substrates. The mechanical characterization performed through small punch tests has evidenced that inclusion of 10 per cent by weight of FeHA would represent an effective reinforcement. The inclusion of nanoparticles also improves the hydrophilicity of the substrates as evidenced by the lower values of water contact angle in comparison with those of neat PCL. The results from magnetic measurements confirmed the superparamagnetic character of the nanocomposite substrates, indicated by a very low coercive field, a saturation magnetization strictly proportional to the FeHA content and a strong history dependence in temperature sweeps. Regarding the biological performances, confocal laser scanning microscopy and AlamarBlue assay have provided qualitative and quantitative information on human mesenchymal stem cell adhesion and viability/proliferation, respectively, whereas the obtained ALP/DNA values have shown the ability of the nanocomposite substrates to support osteogenic differentiation. PMID:23303218

  13. Magnetic poly(ε-caprolactone)/iron-doped hydroxyapatite nanocomposite substrates for advanced bone tissue engineering.

    PubMed

    Gloria, A; Russo, T; D'Amora, U; Zeppetelli, S; D'Alessandro, T; Sandri, M; Bañobre-López, M; Piñeiro-Redondo, Y; Uhlarz, M; Tampieri, A; Rivas, J; Herrmannsdörfer, T; Dediu, V A; Ambrosio, L; De Santis, R

    2013-03-01

    In biomedicine, magnetic nanoparticles provide some attractive possibilities because they possess peculiar physical properties that permit their use in a wide range of applications. The concept of magnetic guidance basically spans from drug delivery and hyperthermia treatment of tumours, to tissue engineering, such as magneto-mechanical stimulation/activation of cell constructs and mechanosensitive ion channels, magnetic cell-seeding procedures, and controlled cell proliferation and differentiation. Accordingly, the aim of this study was to develop fully biodegradable and magnetic nanocomposite substrates for bone tissue engineering by embedding iron-doped hydroxyapatite (FeHA) nanoparticles in a poly(ε-caprolactone) (PCL) matrix. X-ray diffraction analyses enabled the demonstration that the phase composition and crystallinity of the magnetic FeHA were not affected by the process used to develop the nanocomposite substrates. The mechanical characterization performed through small punch tests has evidenced that inclusion of 10 per cent by weight of FeHA would represent an effective reinforcement. The inclusion of nanoparticles also improves the hydrophilicity of the substrates as evidenced by the lower values of water contact angle in comparison with those of neat PCL. The results from magnetic measurements confirmed the superparamagnetic character of the nanocomposite substrates, indicated by a very low coercive field, a saturation magnetization strictly proportional to the FeHA content and a strong history dependence in temperature sweeps. Regarding the biological performances, confocal laser scanning microscopy and AlamarBlue assay have provided qualitative and quantitative information on human mesenchymal stem cell adhesion and viability/proliferation, respectively, whereas the obtained ALP/DNA values have shown the ability of the nanocomposite substrates to support osteogenic differentiation. PMID:23303218

  14. Relationships between the anisotropy of longitudinal wave velocity and hydroxyapatite crystallite orientation in bovine cortical bone.

    PubMed

    Yamamoto, Kazufumi; Nakatsuji, Tomohiro; Yaoi, Yuichiro; Yamato, Yu; Yanagitani, Takahiko; Matsukawa, Mami; Yamazaki, Kaoru; Matsuyama, Yukihiro

    2012-03-01

    Quantitative ultrasound (QUS) is now widely used for evaluating bone in vivo, because obtained ultrasonic wave properties directly reflect the visco-elasticity. Bone tissue is composed of minerals like hydroxyapatite (HAp) and a collagen matrix. HAp crystallites orientation is thus one parameter of bone elasticity. In this study, we experimentally investigated the anisotropy of ultrasonic wave velocity and the HAp crystallites orientation in the axial-radial and axial-tangential planes in detail, using cylindrical specimens obtained from the cortical bone of three bovine femurs. Longitudinal bulk wave propagation was investigated by using a conventional ultrasonic pulse system. We used the one cycle of sinusoidal pulse which was emitted from wide band transmitter. The nominal frequency of the pulse was 1MHz. First, we investigated the anisotropy of longitudinal wave velocity, measuring the anisotropy of velocity in two planes using cylindrical specimens obtained from identical bone areas. The wave velocity changed due to the rotation angle, showing the maximum value in the direction a little off the bone axis. Moreover, X-ray pole figure measurements also indicated that there were small tilts in the HAp crystallites orientation from the bone axis. The tilt angles were similar to those of the highest velocity direction. There were good correlations between velocity and HAp crystallites orientation obtained in different directions. However, a comparatively low correlation was found in posterior bone areas, which shows the stronger effects of bone microstructure. In the radial-tangential plane, where the HAp crystallites hardly ever align, weak anisotropy of velocity was found which seemed to depend on the bone microstructure. PMID:22014464

  15. The Foreign Body Giant Cell Cannot Resorb Bone, But Dissolves Hydroxyapatite Like Osteoclasts

    PubMed Central

    ten Harkel, Bas; Schoenmaker, Ton; Picavet, Daisy I.; Davison, Noel L.; de Vries, Teun J.; Everts, Vincent

    2015-01-01

    Foreign body multinucleated giant cells (FBGCs) and osteoclasts share several characteristics, like a common myeloid precursor cell, multinuclearity, expression of tartrate-resistant acid phosphatase (TRAcP) and dendritic cell-specific transmembrane protein (DC-STAMP). However, there is an important difference: osteoclasts form and reside in the vicinity of bone, while FBGCs form only under pathological conditions or at the surface of foreign materials, like medical implants. Despite similarities, an important distinction between these cell types is that osteoclasts can resorb bone, but it is unknown whether FBGCs are capable of such an activity. To investigate this, we differentiated FBGCs and osteoclasts in vitro from their common CD14+ monocyte precursor cells, using different sets of cytokines. Both cell types were cultured on bovine bone slices and analyzed for typical osteoclast features, such as bone resorption, presence of actin rings, formation of a ruffled border, and characteristic gene expression over time. Additionally, both cell types were cultured on a biomimetic hydroxyapatite coating to discriminate between bone resorption and mineral dissolution independent of organic matrix proteolysis. Both cell types differentiated into multinucleated cells on bone, but FBGCs were larger and had a higher number of nuclei compared to osteoclasts. FBGCs were not able to resorb bone, yet they were able to dissolve the mineral fraction of bone at the surface. Remarkably, FBGCs also expressed actin rings, podosome belts and sealing zones—cytoskeletal organization that is considered to be osteoclast-specific. However, they did not form a ruffled border. At the gene expression level, FBGCs and osteoclasts expressed similar levels of mRNAs that are associated with the dissolution of mineral (e.g., anion exchange protein 2 (AE2), carbonic anhydrase 2 (CAII), chloride channel 7 (CIC7), and vacuolar-type H+-ATPase (v-ATPase)), in contrast the matrix degrading enzyme

  16. Hollow hydroxyapatite microspheres: a novel bioactive and osteoconductive carrier for controlled release of bone morphogenetic protein-2 in bone regeneration

    PubMed Central

    Xiao, Wei; Fu, Hailuo; Rahaman, Mohamed N.; Liu, Yonxing; Bal, B. Sonny

    2013-01-01

    The regeneration of large bone defects is a common and significant clinical problem. Limitations associated with existing treatments such as autologous bone grafts and allografts have increased the need for synthetic bone graft substitutes. The objective of this study was to evaluate the capacity of novel hollow hydroxyapatite (HA) microspheres to serve as a carrier for controlled release of bone morphogenetic-2 (BMP2) in bone regeneration. Hollow HA microspheres (106–150 μm) with a high surface area (>100 m2/g) and a mesoporous shell wall (pore size 10–20 nm) were created using a glass conversion technique. The release of BMP2 from the microspheres into a medium composed of diluted fetal bovine serum in vitro was slow, but it occurred continuously for over 2 weeks. When implanted in rat calvarial defects for 3 or 6 weeks, the microspheres loaded with BMP2 (1 μg/defect) showed a significantly better capacity to regenerate bone than those without BMP2. The amount of new bone in the defects implanted with the BMP2-loaded microspheres was 40% and 43%, respectively, at 3 and 6 weeks, compared to 13% and 17%, respectively, for the microspheres without BMP2. Coating the BMP2-loaded microspheres with a biodegradable polymer, poly(lactic-co-glycolic acid), reduced the amount of BMP2 released in vitro and, above a certain coating thickness, significantly reduced bone regeneration in vivo. The results indicate that these hollow HA microspheres could provide a bioactive and osteoconductive carrier for growth factors in bone regeneration. PMID:23747325

  17. Hydroxyapatite fiber reinforced poly(alpha-hydroxy ester) foams for bone regeneration

    NASA Technical Reports Server (NTRS)

    Thomson, R. C.; Yaszemski, M. J.; Powers, J. M.; Mikos, A. G.; McIntire, L. V. (Principal Investigator)

    1998-01-01

    A process has been developed to manufacture biodegradable composite foams of poly(DL-lactic-co-glycolic acid) (PLGA) and hydroxyapatite short fibers for use in bone regeneration. The processing technique allows the manufacture of three-dimensional foam scaffolds and involves the formation of a composite material consisting of a porogen material (either gelatin microspheres or salt particles) and hydroxyapatite short fibers embedded in a PLGA matrix. After the porogen is leached out, an open-cell composite foam remains which has a pore size and morphology defined by the porogen. By changing the weight fraction of the leachable component it was possible to produce composite foams with controlled porosities ranging from 0.47 +/- 0.02 to 0.85 +/- 0.01 (n = 3). Up to a polymer:fiber ratio of 7:6, short hydroxyapatite fibers served to reinforce low-porosity PLGA foams manufactured using gelatin microspheres as a porogen. Foams with a compressive yield strength up to 2.82 +/- 0.63 MPa (n = 3) and a porosity of 0.47 +/- 0.02 (n = 3) were manufactured using a polymer:fiber weight ratio of 7:6. In contrast, high-porosity composite foams (up to 0.81 +/- 0.02, n = 3) suitable for cell seeding were not reinforced by the introduction of increasing quantities of hydroxyapatite short fibers. We were therefore able to manufacture high-porosity foams which may be seeded with cells but which have minimal compressive yield strength, or low porosity foams with enhanced osteoconductivity and compressive yield strength.

  18. Development of hydroxyapatite scaffolds for artificial bone grafts

    NASA Astrophysics Data System (ADS)

    Ajaal, Tawfik Taher

    The Taguchi method of experimental design is very well suited to improving the production process of the synthetic bone grafts for several reasons. First, it is very efficient and easy to apply, so that it does not require large amounts of time or resources to conduct a given set of experiments. This makes it possible to conduct a series of experiments that result in continuous process improvement. Second, the effect of many different process variables can be examined simultaneously, which ensures that beneficial factor combinations are not overlooked. Finally, using a Taguchi signal-to-noise ratio leads to concurrent optimization of the process and reduction of process variability. The application of Taguchi method was successful in optimizing the production process of the synthetic bone grafts. The compressive strength was doubled while maintaining the appropriate porosity level and microstructure for the bioactivity process. The mean value of the compression strength obtained was 5.8 MPa with density of 0.515 gm/cm3 for samples prepared from 45 pores per inch (PPI) foam reticulate, and 3.2 MPa with 0.422 gm/cm 3 for samples prepared from 30 PPI foam reticulate. Three levels of porosity were identified namely, macro, meso, and micro-porosity. The pore sizes were (350--400) mum, (100--120) mum and (2--6) mum respectively based on the used substrate. Using the Taguchi method in conjunction with a statistical experimental design, the various steps of the scaffold production process such as slurry preparation, coating process, drying, calcining and sintering processes were optimized. The final optimized process gave highly reproducible results.

  19. [FTIR and XRD analysis of hydroxyapatite from fossil human and animal teeth in Jinsha Relict, Chengdu].

    PubMed

    Huang, Cheng-min; Zhang, Qing; Bai, Song; Wang, Cheng-shan

    2007-12-01

    Diagenetic effect during burial on the hydroxyapatite in enamel and dentin from fossil human and animal teeth was examined, using X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). For the enamel and dentin of all fossil teeth, XRD patterns exhibit bulge line and overlap between major reflections of hydroxyapatite, and the crystallinity of hydroxyapatite is low. For each infrared spectrum, H2O and OH(-) have distinct peaks of absorbance, and PO4(3-) and CO3(2-) ions have intensive infrared vibration modes at the fundamental wave numbers. The component of hydroxyapatite of all fossil teeth is similar to the modern biological hydroxyapatite. Furthermore, the index (PCI) which reflects the hydroxyapatite crystallinity of each sample ranges from 2.4 to 4.0 while the index (BPI) reflecting the amount of type B carbonate to phosphate indicates that the values of CO3(2-) content in hydroxyapatite are rather high, accordingly the crystallinity of all fossil hydroxyapatites are poor. It could be concluded that little alteration of hydroxyapatites from fossil human and animal teeth occurred in the process of diagenesis in Jinsha Relict, Chengdu, China. PMID:18330282

  20. Facilitated receptor-recognition and enhanced bioactivity of bone morphogenetic protein-2 on magnesium-substituted hydroxyapatite surface

    NASA Astrophysics Data System (ADS)

    Huang, Baolin; Yuan, Yuan; Li, Tong; Ding, Sai; Zhang, Wenjing; Gu, Yuantong; Liu, Changsheng

    2016-04-01

    Biomaterial surface functionalized with bone morphogenetic protein-2 (BMP-2) is a promising approach to fabricating successful orthopedic implants/scaffolds. However, the bioactivity of BMP-2 on material surfaces is still far from satisfactory and the mechanism of related protein-surface interaction remains elusive. Based on the most widely used bone-implants/scaffolds material, hydroxyapatite (HAP), we developed a matrix of magnesium-substituted HAP (Mg-HAP, 2.2 at% substitution) to address these issues. Further, we investigated the adsorption dynamics, BMPRs-recruitment, and bioactivity of recombinant human BMP-2 (rhBMP-2) on the HAP and Mg-HAP surfaces. To elucidate the mechanism, molecular dynamic simulations were performed to calculate the preferred orientations, conformation changes, and cysteine-knot stabilities of adsorbed BMP-2 molecules. The results showed that rhBMP-2 on the Mg-HAP surface exhibited greater bioactivity, evidenced by more facilitated BMPRs-recognition and higher ALP activity than on the HAP surface. Moreover, molecular simulations indicated that BMP-2 favoured distinct side-on orientations on the HAP and Mg-HAP surfaces. Intriguingly, BMP-2 on the Mg-HAP surface largely preserved the active protein structure evidenced by more stable cysteine-knots than on the HAP surface. These findings explicitly clarify the mechanism of BMP-2-HAP/Mg-HAP interactions and highlight the promising application of Mg-HAP/BMP-2 matrixes in bone regeneration implants/scaffolds.

  1. Facilitated receptor-recognition and enhanced bioactivity of bone morphogenetic protein-2 on magnesium-substituted hydroxyapatite surface

    PubMed Central

    Huang, Baolin; Yuan, Yuan; Li, Tong; Ding, Sai; Zhang, Wenjing; Gu, Yuantong; Liu, Changsheng

    2016-01-01

    Biomaterial surface functionalized with bone morphogenetic protein-2 (BMP-2) is a promising approach to fabricating successful orthopedic implants/scaffolds. However, the bioactivity of BMP-2 on material surfaces is still far from satisfactory and the mechanism of related protein-surface interaction remains elusive. Based on the most widely used bone-implants/scaffolds material, hydroxyapatite (HAP), we developed a matrix of magnesium-substituted HAP (Mg-HAP, 2.2 at% substitution) to address these issues. Further, we investigated the adsorption dynamics, BMPRs-recruitment, and bioactivity of recombinant human BMP-2 (rhBMP-2) on the HAP and Mg-HAP surfaces. To elucidate the mechanism, molecular dynamic simulations were performed to calculate the preferred orientations, conformation changes, and cysteine-knot stabilities of adsorbed BMP-2 molecules. The results showed that rhBMP-2 on the Mg-HAP surface exhibited greater bioactivity, evidenced by more facilitated BMPRs-recognition and higher ALP activity than on the HAP surface. Moreover, molecular simulations indicated that BMP-2 favoured distinct side-on orientations on the HAP and Mg-HAP surfaces. Intriguingly, BMP-2 on the Mg-HAP surface largely preserved the active protein structure evidenced by more stable cysteine-knots than on the HAP surface. These findings explicitly clarify the mechanism of BMP-2-HAP/Mg-HAP interactions and highlight the promising application of Mg-HAP/BMP-2 matrixes in bone regeneration implants/scaffolds. PMID:27075233

  2. Scaffolds for bone regeneration made of hydroxyapatite microspheres in a collagen matrix.

    PubMed

    Cholas, Rahmatullah; Kunjalukkal Padmanabhan, Sanosh; Gervaso, Francesca; Udayan, Gayatri; Monaco, Graziana; Sannino, Alessandro; Licciulli, Antonio

    2016-06-01

    Biomimetic scaffolds with a structural and chemical composition similar to native bone tissue may be promising for bone tissue regeneration. In the present work hydroxyapatite mesoporous microspheres (mHA) were incorporated into collagen scaffolds containing an ordered interconnected macroporosity. The mHA were obtained by spray drying of a nano hydroxyapatite slurry prepared by the precipitation technique. X-ray diffraction (XRD) analysis revealed that the microspheres were composed only of hydroxyapatite (HA) phase, and energy-dispersive x-ray spectroscopy (EDS) analysis revealed the Ca/P ratio to be 1.69 which is near the value for pure HA. The obtained microspheres had an average diameter of 6μm, a specific surface area of 40m(2)/g as measured by Brunauer-Emmett-Teller (BET) analysis, and Barrett-Joyner-Halenda (BJH) analysis showed a mesoporous structure with an average pore diameter of 16nm. Collagen/HA-microsphere (Col/mHA) composite scaffolds were prepared by freeze-drying followed by dehydrothermal crosslinking. SEM observations of Col/mHA scaffolds revealed HA microspheres embedded within a porous collagen matrix with a pore size ranging from a few microns up to 200μm, which was also confirmed by histological staining of sections of paraffin embedded scaffolds. The compressive modulus of the composite scaffold at low and high strain values was 1.7 and 2.8 times, respectively, that of pure collagen scaffolds. Cell proliferation measured by the MTT assay showed more than a 3-fold increase in cell number within the scaffolds after 15days of culture for both pure collagen scaffolds and Col/mHA composite scaffolds. Attractive properties of this composite scaffold include the potential to load the microspheres for drug delivery and the controllability of the pore structure at various length scales. PMID:27040244

  3. The role of collagen in bone apatite formation in the presence of hydroxyapatite nucleation inhibitors

    PubMed Central

    Nudelman, Fabio; Pieterse, Koen; George, Anne; Bomans, Paul H. H.; Friedrich, Heiner; Brylka, Laura J.; Hilbers, Peter A. J.; de With, Gijsbertus; Sommerdijk, Nico A. J. M.

    2011-01-01

    Bone is a composite material, in which collagen fibrils form a scaffold for a highly organized arrangement of uniaxially oriented apatite crystals1,2. In the periodic 67 nm cross-striated pattern of the collagen fibril3–5, the less dense 40-nm-long gap zone has been implicated as the place where apatite crystals nucleate from an amorphous phase, and subsequently grow6–9. This process is believed to be directed by highly acidic non-collagenous proteins6,7,9–11; however, the role of the collagen matrix12–14 during bone apatite mineralization remains unknown. Here, combining nanometre-scale resolution cryogenic transmission electron microscopy and cryogenic electron tomography15 with molecular modelling, we show that collagen functions in synergy with inhibitors of hydroxyapatite nucleation to actively control mineralization. The positive net charge close to the C-terminal end of the collagen molecules promotes the infiltration of the fibrils with amorphous calcium phosphate (ACP). Furthermore, the clusters of charged amino acids, both in gap and overlap regions, form nucleation sites controlling the conversion of ACP into a parallel array of oriented apatite crystals. We developed a model describing the mechanisms through which the structure, supramolecular assembly and charge distribution of collagen can control mineralization in the presence of inhibitors of hydroxyapatite nucleation. PMID:20972429

  4. Three-dimensional printed PCL-hydroxyapatite scaffolds filled with CNTs for bone cell growth stimulation.

    PubMed

    Gonçalves, Elsa M; Oliveira, Filipe J; Silva, Rui F; Neto, Miguel A; Fernandes, M Helena; Amaral, Margarida; Vallet-Regí, María; Vila, Mercedes

    2016-08-01

    A three-phase [nanocrystalline hydroxyapatite (HA), carbon nanotubes (CNT), mixed in a polymeric matrix of polycaprolactone (PCL)] composite scaffold produced by 3D printing is presented. The CNT content varied between 0 and 10 wt % in a 50 wt % PCL matrix, with HA being the balance. With the combination of three well-known materials, these scaffolds aimed at bringing together the properties of all into a unique material to be used in tissue engineering as support for cell growth. The 3D printing technique allows producing composite scaffolds having an interconnected network of square pores in the range of 450-700 μm. The 2 wt % CNT scaffold offers the best combination of mechanical behaviour and electrical conductivity. Its compressive strength of ∼4 MPa is compatible with the trabecular bone. The composites show typical hydroxyapatite bioactivity, good cell adhesion and spreading at the scaffolds surface, this combination of properties indicating that the produced 3D, three-phase, scaffolds are promising materials in the field of bone regenerative medicine. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1210-1219, 2016. PMID:26089195

  5. Reduced graphene oxide-coated hydroxyapatite composites stimulate spontaneous osteogenic differentiation of human mesenchymal stem cells

    NASA Astrophysics Data System (ADS)

    Lee, Jong Ho; Shin, Yong Cheol; Jin, Oh Seong; Kang, Seok Hee; Hwang, Yu-Shik; Park, Jong-Chul; Hong, Suck Won; Han, Dong-Wook

    2015-07-01

    Human mesenchymal stem cells (hMSCs) have great potential as cell sources for bone tissue engineering and regeneration, but the control and induction of their specific differentiation into bone cells remain challenging. Graphene-based nanomaterials are considered attractive candidates for biomedical applications such as scaffolds in tissue engineering, substrates for SC differentiation and components of implantable devices, due to their biocompatible and bioactive properties. Despite the potential biomedical applications of graphene and its derivatives, only limited information is available regarding their osteogenic activity. This study concentrates upon the effects of reduced graphene oxide (rGO)-coated hydroxyapatite (HAp) composites on osteogenic differentiation of hMSCs. The average particle sizes of HAp and rGO were 1270 +/- 476 nm and 438 +/- 180 nm, respectively. When coated on HAp particulates, rGO synergistically enhanced spontaneous osteogenic differentiation of hMSCs, without hampering their proliferation. This result was confirmed by determining alkaline phosphatase activity and mineralization of calcium and phosphate as early and late stage markers of osteogenic differentiation. It is suggested that rGO-coated HAp composites can be effectively utilized as dental and orthopedic bone fillers since these graphene-based particulate materials have potent effects on stimulating the spontaneous differentiation of MSCs and show superior bioactivity and osteoinductive potential.Human mesenchymal stem cells (hMSCs) have great potential as cell sources for bone tissue engineering and regeneration, but the control and induction of their specific differentiation into bone cells remain challenging. Graphene-based nanomaterials are considered attractive candidates for biomedical applications such as scaffolds in tissue engineering, substrates for SC differentiation and components of implantable devices, due to their biocompatible and bioactive properties. Despite

  6. Histomorphometric evaluation of strontium-containing nanostructured hydroxyapatite as bone substitute in sheep.

    PubMed

    Machado, Callinca Paolla Gomes; Sartoretto, Suelen Cristina; Alves, Adriana Terezinha Neves Novellino; Lima, Inayá Barbosa Correa; Rossi, Alexandre Malta; Granjeiro, José Mauro; Calasans-Maia, Mônica Diuana

    2016-01-01

    The aim of this study is to evaluate the biocompatibility and osteoconductivity in surgical defects of sheep tibias filled with 1% strontium-containing nanostructured hydroxyapatite microspheres (SrHA), stoichiometric hydroxyapatite without strontium microspheres (HA), or blood clots. Santa Ines sheep were subjected to three perforations on the medial side of the left tibia. The biomaterials were characterized by X-ray Diffraction (XRD) and Fourier Transform Infrared (FTIR) before implantation and by X-Ray Microfluorescence (µFRX) and Scanning Electron Microscopy (SEM) after sheep tibias implantation. Surgical defects were filled with blood clots (control), SrHA (Group 1) or HA (Group 2). After 30 days, 5-µm bone blocks were obtained for histological evaluation, and the blocks obtained from 1 animal were embedded in methylmethacrylate for undecalcified sections. Mononuclear inflammatory infiltrate remained mild in all experimental groups. Giant cells were observed surrounding biomaterials particles of both groups and areas of bone formation were detected in close contact with biomaterials. All groups showed newly formed bone from the periphery to the center of the defects, which the control, HA and SrHA presented 36.4% (± 21.8), 31.2% (± 14.7) and 26.2% (± 12.9) of newly formed bone density, respectively, not presenting statistical differences. In addition, the connective tissue density did not show any significant between groups. The SrHA showing a higher volume density of biomaterial (51.2 ± 14.1) present in the defect compared to HA (32.6 ± 8.5) after 30 days (p = 0.03). Microspheres containing 1% SrHA or HA can be considered biocompatible, have osteoconductive properties and may be useful biomaterials for clinical applications. PMID:27191738

  7. Biodegradation and cytotoxicity of ciprofloxacin-loaded hydroxyapatite-polycaprolactone nanocomposite film for sustainable bone implants

    PubMed Central

    Nithya, Rajendran; Meenakshi Sundaram, Nachiappan

    2015-01-01

    Introduction In recent years there has been a steep increase in the number of orthopedic patients for many reasons. One major reason is osteomyelitis, caused by pyrogenic bacteria, with progressive infection of the bone or bone marrow and surrounding tissues. So antibiotics must be introduced during bone implantation to avoid prolonged infection. Aim The objective of the study reported here was to prepare a composite film of nanocrystalline hydroxyapatite (HAp) and polycaprolactone (PCL) polymer loaded with ciprofloxacin, a frequently used antibiotic agent for bone infections. Methods Nanocrystalline HAp was synthesized by precipitation method using the precursor obtained from eggshell. The nanocomposite film (HAp-PCL-ciprofloxacin) was prepared by solvent evaporation. Drug-release and biodegradation studies were undertaken by immersing the composite film in phosphate-buffered saline solution, while a cytotoxicity test was performed using the fibroblast cell line NIH-3T3 and osteoblast cell line MG-63. Results The pure PCL film had quite a low dissolution rate after an initial sharp weight loss, whereas the ciprofloxacin-loaded HAp-PCL nanocomposite film had a large weight loss due to its fast drug release. The composite film had higher water absorption than the pure PCL, and increasing the concentration of the HAp increased the water absorption. The in vitro cell-line study showed a good biocompatibility and bioactivity of the developed nanocomposite film. Conclusion The prepared film will act as a sustainable bone implant in addition to controlled drug delivery. PMID:26491313

  8. Greener synthesis of electrospun collagen/hydroxyapatite composite fibers with an excellent microstructure for bone tissue engineering

    PubMed Central

    Zhou, Yuanyuan; Yao, Hongchang; Wang, Jianshe; Wang, Dalu; Liu, Qian; Li, Zhongjun

    2015-01-01

    In bone tissue engineering, collagen/hydroxyapatite (HAP) fibrous composite obtained via electrospinning method has been demonstrated to support the cells’ adhesion and bone regeneration. However, electrospinning of natural collagen often requires the use of cytotoxic organic solvents, and the HAP crystals were usually aggregated and randomly distributed within a fibrous matrix of collagen, limiting their clinical potential. Here, an effective and greener method for the preparation of collagen/HAP composite fibers was developed for the first time, and this green product not only had 40 times higher mechanical properties than that previously reported, but also had an excellent microstructure similar to that of natural bone. By dissolving type I collagen in environmentally friendly phosphate buffered saline/ethanol solution instead of the frequently-used cytotoxic organic solvents, followed with the key step of desalination, co-electrospinning the collagen solution with the HAP sol, generates a collagen/HAP composite with a uniform and continuous fibrous morphology. Interestingly, the nano-HAP needles were found to preferentially orient along the longitudinal direction of the collagen fibers, which mimicked the nanostructure of natural bones. Based on the characterization of the related products, the formation mechanism for this novel phenomenon was proposed. After cross-linking with 1-ethyl-3-(3-dimethyl-aminopropyl)-1-carbodiimide hydrochloride/N-hydroxysuccinimide, the obtained composite exhibited a significant enhancement in mechanical properties. In addition, the biocompatibility of the obtained composite fibers was evaluated by in vitro culture of the human myeloma cells (U2-OS). Taken together, the process outlined herein provides an effective, non-toxic approach for the fabrication of collagen/HAP composite nanofibers that could be good candidates for bone tissue engineering. PMID:25995630

  9. Greener synthesis of electrospun collagen/hydroxyapatite composite fibers with an excellent microstructure for bone tissue engineering.

    PubMed

    Zhou, Yuanyuan; Yao, Hongchang; Wang, Jianshe; Wang, Dalu; Liu, Qian; Li, Zhongjun

    2015-01-01

    In bone tissue engineering, collagen/hydroxyapatite (HAP) fibrous composite obtained via electrospinning method has been demonstrated to support the cells' adhesion and bone regeneration. However, electrospinning of natural collagen often requires the use of cytotoxic organic solvents, and the HAP crystals were usually aggregated and randomly distributed within a fibrous matrix of collagen, limiting their clinical potential. Here, an effective and greener method for the preparation of collagen/HAP composite fibers was developed for the first time, and this green product not only had 40 times higher mechanical properties than that previously reported, but also had an excellent microstructure similar to that of natural bone. By dissolving type I collagen in environmentally friendly phosphate buffered saline/ethanol solution instead of the frequently-used cytotoxic organic solvents, followed with the key step of desalination, co-electrospinning the collagen solution with the HAP sol, generates a collagen/HAP composite with a uniform and continuous fibrous morphology. Interestingly, the nano-HAP needles were found to preferentially orient along the longitudinal direction of the collagen fibers, which mimicked the nanostructure of natural bones. Based on the characterization of the related products, the formation mechanism for this novel phenomenon was proposed. After cross-linking with 1-ethyl-3-(3-dimethyl-aminopropyl)-1-carbodiimide hydrochloride/N-hydroxysuccinimide, the obtained composite exhibited a significant enhancement in mechanical properties. In addition, the biocompatibility of the obtained composite fibers was evaluated by in vitro culture of the human myeloma cells (U2-OS). Taken together, the process outlined herein provides an effective, non-toxic approach for the fabrication of collagen/HAP composite nanofibers that could be good candidates for bone tissue engineering. PMID:25995630

  10. Biomimetic fiber mesh scaffolds based on gelatin and hydroxyapatite nano-rods: Designing intrinsic skills to attain bone reparation abilities.

    PubMed

    Sartuqui, Javier; Gravina, A Noel; Rial, Ramón; Benedini, Luciano A; Yahia, L'Hocine; Ruso, Juan M; Messina, Paula V

    2016-09-01

    Intrinsic material skills have a deep effect on the mechanical and biological performance of bone substitutes, as well as on its associated biodegradation properties. In this work we have manipulated the preparation of collagenous derived fiber mesh frameworks to display a specific composition, morphology, open macroporosity, surface roughness and permeability characteristics. Next, the effect of the induced physicochemical attributes on the scaffold's mechanical behavior, bone bonding potential and biodegradability were evaluated. It was found that the scaffold microstructure, their inherent surface roughness, and the compression strength of the gelatin scaffolds can be modulated by the effect of the cross-linking agent and, essentially, by mimicking the nano-scale size of hydroxyapatite in natural bone. A clear effect of bioactive hydroxyapatite nano-rods on the scaffolds skills can be appreciated and it is greater than the effect of the cross-linking agent, offering a huge perspective for the upcoming progress of bone implant technology. PMID:27220014

  11. Selective laser sintering fabrication of nano-hydroxyapatite/poly-ε-caprolactone scaffolds for bone tissue engineering applications.

    PubMed

    Xia, Yan; Zhou, Panyu; Cheng, Xiaosong; Xie, Yang; Liang, Chong; Li, Chao; Xu, Shuogui

    2013-01-01

    The regeneration of functional tissue in osseous defects is a formidable challenge in orthopedic surgery. In the present study, a novel biomimetic composite scaffold, here called nano-hydroxyapatite (HA)/poly-ε-caprolactone (PCL) was fabricated using a selective laser sintering technique. The macrostructure, morphology, and mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the nano-HA/PCL scaffolds exhibited predesigned, well-ordered macropores and interconnected micropores. The scaffolds have a range of porosity from 78.54% to 70.31%, and a corresponding compressive strength of 1.38 MPa to 3.17 MPa. Human bone marrow stromal cells were seeded onto the nano-HA/PCL or PCL scaffolds and cultured for 28 days in vitro. As indicated by the level of cell attachment and proliferation, the nano-HA/PCL showed excellent biocompatibility, comparable to that of PCL scaffolds. The hydrophilicity, mineralization, alkaline phosphatase activity, and Alizarin Red S staining indicated that the nano-HA/PCL scaffolds are more bioactive than the PCL scaffolds in vitro. Measurements of recombinant human bone morphogenetic protein-2 (rhBMP-2) release kinetics showed that after nano-HA was added, the material increased the rate of rhBMP-2 release. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both nano-HA/PCL scaffolds and PCL scaffolds were implanted in rabbit femur defects for 3, 6, and 9 weeks. The wounds were studied radiographically and histologically. The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility. However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds. Thus, they show large potential for use in orthopedic and reconstructive surgery. PMID:24204147

  12. Selective laser sintering fabrication of nano-hydroxyapatite/poly-ε-caprolactone scaffolds for bone tissue engineering applications

    PubMed Central

    Xia, Yan; Zhou, Panyu; Cheng, Xiaosong; Xie, Yang; Liang, Chong; Li, Chao; Xu, Shuogui

    2013-01-01

    The regeneration of functional tissue in osseous defects is a formidable challenge in orthopedic surgery. In the present study, a novel biomimetic composite scaffold, here called nano-hydroxyapatite (HA)/poly-ε-caprolactone (PCL) was fabricated using a selective laser sintering technique. The macrostructure, morphology, and mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the nano-HA/PCL scaffolds exhibited predesigned, well-ordered macropores and interconnected micropores. The scaffolds have a range of porosity from 78.54% to 70.31%, and a corresponding compressive strength of 1.38 MPa to 3.17 MPa. Human bone marrow stromal cells were seeded onto the nano-HA/PCL or PCL scaffolds and cultured for 28 days in vitro. As indicated by the level of cell attachment and proliferation, the nano-HA/PCL showed excellent biocompatibility, comparable to that of PCL scaffolds. The hydrophilicity, mineralization, alkaline phosphatase activity, and Alizarin Red S staining indicated that the nano-HA/PCL scaffolds are more bioactive than the PCL scaffolds in vitro. Measurements of recombinant human bone morphogenetic protein-2 (rhBMP-2) release kinetics showed that after nano-HA was added, the material increased the rate of rhBMP-2 release. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both nano-HA/PCL scaffolds and PCL scaffolds were implanted in rabbit femur defects for 3, 6, and 9 weeks. The wounds were studied radiographically and histologically. The in vivo results showed that both nano-HA/PCL composite scaffolds and PCL scaffolds exhibited good biocompatibility. However, the nano-HA/PCL scaffolds enhanced the efficiency of new bone formation more than PCL scaffolds and fulfilled all the basic requirements of bone tissue engineering scaffolds. Thus, they show large potential for use in orthopedic and reconstructive surgery. PMID:24204147

  13. Matched Comparison of Fusion Rates between Hydroxyapatite Demineralized Bone Matrix and Autograft in Lumbar Interbody Fusion

    PubMed Central

    Kim, Dae Hwan; Lee, Nam; Shin, Dong Ah; Yi, Seong; Kim, Keung Nyun

    2016-01-01

    Objective To compare the fusion rate of a hydroxyapatite demineralized bone matrix (DBM) with post-laminectomy acquired autograft in lumbar interbody fusion surgery and to evaluate the correlation between fusion rate and clinical outcome. Methods From January 2013 to April 2014, 98 patients underwent lumbar interbody fusion surgery with hydroxyapatite DBM (HA-DBM group) in our institute. Of those patients, 65 received complete CT scans for 12 months postoperatively in order to evaluate fusion status. For comparison with autograft, we selected another 65 patients who underwent lumbar interbody fusion surgery with post-laminectomy acquired autograft (Autograft group) during the same period. Both fusion material groups were matched in terms of age, sex, body mass index (BMI), and bone mineral density (BMD). To evaluate the clinical outcomes, we analyzed the results of visual analogue scale (VAS), Oswestry Disability Index (ODI), and Short Form Health Survey (SF-36). Results We reviewed the CT scans of 149 fusion levels in 130 patients (HA-DBM group, 75 levels/65 patients; Autograft group, 74 levels/65 patients). Age, sex, BMI, and BMD were not significantly different between the groups (p=0.528, p=0.848, p=0.527, and p=0.610, respectively). The HA-DBM group showed 39 of 75 fused levels (52%), and the Autograft group showed 46 of 74 fused levels (62.2%). This difference was not statistically significant (p=0.21). In the HA-DBM group, older age and low BMD were significantly associated with non-fusion (61.24 vs. 66.68, p=0.027; -1.63 vs. -2.29, p=0.015, respectively). VAS and ODI showed significant improvement after surgery when fusion was successfully achieved in both groups (p=0.004, p=0.002, HA-DBM group; p=0.012, p=0.03, Autograft group). Conclusion The fusion rates of the hydroxyapatite DBM and Autograft groups were not significantly different. In addition, clinical outcomes were similar between the groups. However, older age and low BMD are risk factors that might

  14. Poly-L-lactic acid/hydroxyapatite electrospun nanocomposites induce chondrogenic differentiation of human MSC.

    PubMed

    Spadaccio, Cristiano; Rainer, Alberto; Trombetta, Marcella; Vadalá, Gianluca; Chello, Massimo; Covino, Elvio; Denaro, Vincenzo; Toyoda, Yoshiya; Genovese, Jorge A

    2009-07-01

    Cartilage and bone tissue engineering has been widely investigated but is still hampered by cell differentiation and transplant integration issues within the constructs. Scaffolds represent the pivotal structure of the engineered tissue and establish an environment for neo-extracellular matrix synthesis. They can be associated to signals to modulate cell activity. In this study, considering the well reported role of hydroxyapatite (HA) in cartilage repair, we focused on the putative chondrogenic differentiation of human mesenchymal stem cells (hMSCs) following culture on membranes of electrospun fibers of poly-L-lactic acid (PLLA) loaded with nanoparticles of HA. hMSCs were seeded on PLLA/HA and bare PLLA membranes and cultured in basal medium, using chondrogenic differentiation medium as a positive control. After 14 days of culture, SOX-9 positive cells could be detected in the PLLA/HA group. Cartilage specific proteoglycan immunostain confirmed the presence of neo-extracellular-matrix production. Co-expression of CD29, a typical surface marker of MSCs and SOX-9, suggested different degrees in the differentiation process. We developed a hydroxyapatite functionalized scaffold with the aim to recapitulate the native histoarchitecture and the molecular signaling of osteochondral tissue to facilitate cell differentiation toward chondrocyte. PLLA/HA nanocomposites induced differentiation of hMSCs in a chondrocyte-like phenotype with generation of a proteoglycan based matrix. This nanocomposite could be an amenable alternative scaffold for cartilage tissue engineering using hMSCs. PMID:19418224

  15. Dynamic photophysical processes in laser irradiated human cortical skull bone

    NASA Astrophysics Data System (ADS)

    Mandelis, Andreas; Kwan, Chi-Hang; Matvienko, Anna

    2009-02-01

    Modulated luminescence (LUM) technique was applied to analyze photophysical processes in the cortical layer of human skull bones. The theoretical interpretation of the results was based on the optical excitation and decay rate equations of the fluorophore and on the molecular interaction parameter with the photon field density in the matrix of the bone. Using comparisons of the theory with the frequency response of dental LUM it was concluded that the optically active molecular species (fluorophore) in the bones is hydroxyapatite. An effective relaxation lifetime of skull cortical bone was derived theoretically and was found to depend on the intrinsic fluorophore decay lifetime, on the photon field density, and on the thickness of the bone. The experimentally measured dependencies were in excellent agreement with the theoretical model. The theory was able to yield measurements of the optical scattering coefficient, optical absorption coefficient, and mean coupling coefficient. These results show that the quantitative LUM can be used as a sensitive method to measure optical properties of the active fluorophore in cortical skull bones and the optical-field-induced molecular interaction parameter. When calibrated vs. laser intensity, the modulated luminescence can also be used to measure human skull thickness. These traits can be applied to monitor the bone mineral density (BMD) and, ultimately can be used as potential markers of bone health or disease, such as osteoporosis or bone cancer.

  16. Evaluation of human osteoblastic cell response to plasma-sprayed silicon-substituted hydroxyapatite coatings over titanium substrates.

    PubMed

    Gomes, Pedro S; Botelho, Cláudia; Lopes, Maria A; Santos, José D; Fernandes, Maria H

    2010-08-01

    Silicon-substituted hydroxyapatite (Si-HA) coatings have been plasma sprayed over titanium substrates (Ti-6Al-4V) aiming to improve the bioactivity of the constructs for bone tissue repair/regeneration. X-ray diffraction analysis of the coatings has shown that, previous to the thermal deposition, no secondary phases were formed due to the incorporation of 0.8 wt % Si into HA crystal lattice. Partial decomposition of hydroxyapatite, which lead to the formation of the more soluble phases of alpha- and beta-tricalcium phosphate and calcium oxide, and increase of amorphization level only occurred following plasma spraying. Human bone marrow-derived osteoblastic cells were used to assess the in vitro biocompatibility of the constructs. Cells attached and grew well on the Si-HA coatings, putting in evidence an increased metabolic activity and alkaline phosphatase expression comparing to control, i.e., titanium substrates plasma sprayed with hydroxyapatite. Further, a trend for increased differentiation was also verified by the upregulation of osteogenesis-related genes, as well as by the augmented deposition of globular mineral deposits within established cell layers. Based on the present findings, plasma spraying of Si-HA coatings over titanium substrates demonstrates improved biological properties regarding cell proliferation and differentiation, comparing to HA coatings. This suggests that incorporation of Si into the HA lattice could enhance the biological behavior of the plasma-sprayed coating. PMID:20574971

  17. Three-dimensional polycaprolactone-hydroxyapatite scaffolds combined with bone marrow cells for cartilage tissue engineering.

    PubMed

    Wei, Bo; Yao, Qingqiang; Guo, Yang; Mao, Fengyong; Liu, Shuai; Xu, Yan; Wang, Liming

    2015-08-01

    The goal of this study was to investigate the chondrogenic potential of three-dimensional polycaprolactone-hydroxyapatite (PCL-HA) scaffolds loaded with bone marrow cells in vitro and the effect of PCL-HA scaffolds on osteochondral repair in vivo. Here, bone marrow was added to the prepared PCL-HA scaffolds and cultured in chondrogenic medium for 10 weeks. Osteochondral defects were created in the trochlear groove of 29 knees in 17 New Zealand white rabbits, which were then divided into four groups that underwent: implantation of PCL-HA scaffolds (left knee, n = 17; Group 1), microfracture (right knee, n = 6; Group 2), autologous osteochondral transplantation (right knee, n = 6; Group 3), and no treatment (right knee, n = 5; Control). Extracellular matrix produced by bone marrow cells covered the surface and filled the pores of PCL-HA scaffolds after 10 weeks in culture. Moreover, many cell-laden cartilage lacunae were observed, and cartilage matrix was concentrated in the PCL-HA scaffolds. After a 12-week repair period, Group 1 showed excellent vertical and lateral integration with host bone, but incomplete cartilage regeneration and matrix accumulation. An uneven surface of regenerated cartilage and reduced distribution of cartilage matrix were observed in Group 2. In addition, abnormal bone growth and unstable integration between repaired and host tissues were detected. For Group 3, the integration between transplanted and host cartilage was interrupted. Our findings indicate that the PCL-HA scaffolds loaded with bone marrow cells improved chondrogenesis in vitro and implantation of PCL-HA scaffolds for osteochondral repairenhanced integration with host bone. However, cartilage regeneration remained unsatisfactory. The addition of trophic factors or the use of precultured cell-PCL-HA constructs for accelerated osteochondral repair requires further investigation. PMID:25766036

  18. Enhanced bone regeneration with carbon nanotube reinforced hydroxyapatite in animal model.

    PubMed

    Mukherjee, Susmita; Nandi, Samit Kumar; Kundu, Biswanath; Chanda, Abhijit; Sen, Swarnendu; Das, Pradip Kumar

    2016-07-01

    In order to improve the inherently poor mechanical properties of hydroxyapatite (HAp) and to increase its feasibility as load bearing implant material, in the present investigation, functionalised (HFC1 and HFC2) and non-functionalized (HC1 and HC2) multi-walled carbon nanotubes were used as reinforcing material with HAp. Significant improvement with respect to fracture toughness, flexural strength and impact strength of the composites was noticed. In vitro biological properties of HAp-carbon nanotube (CNT) biocomposites have also favored uniform and systematic apatite growth on their surface. Subsequently, in vivo osseous ingrowth at bone defect of rabbit femur was evaluated and compared using radiology, push out test, fluorochrome labeling, histology and scanning electron microscopy after 2 and 4 months respectively. The results demonstrated growth of web like soft callus from the host bone towards the implant, ensuring strong host bone interaction. Toxicological studies of the liver and kidney cells exhibited no abnormality, thereby confirming non-toxicity of the CNT in the animal body. Host-implant biomechanical strength showed high interfacial strength of the composites, indicating their high potentials to be used for bone remodeling applications. PMID:26907099

  19. Engineering nanocages with polyglutamate domains for coupling to hydroxyapatite biomaterials and allograft bone.

    PubMed

    Culpepper, Bonnie K; Morris, David S; Prevelige, Peter E; Bellis, Susan L

    2013-03-01

    Hydroxyapatite (HA) is the principal constituent of bone mineral, and synthetic HA is widely used as a biomaterial for bone repair. Previous work has shown that polyglutamate domains bind selectively to HA and that these domains can be utilized to couple bioactive peptides onto many different HA-containing materials. In the current study we have adapted this technology to engineer polyglutamate domains into cargo-loaded nanocage structures derived from the P22 bacteriophage. P22 nanocages have demonstrated significant potential as a drug delivery system due to their stability, large capacity for loading with a diversity of proteins and other types of cargo, and ability to resist degradation by proteases. Site-directed mutagenesis was used to modify the primary coding sequence of the P22 coat protein to incorporate glutamate-rich regions. Relative to wild-type P22, the polyglutamate-modified nanocages (E2-P22) exhibited increased binding to ceramic HA disks, particulate HA and allograft bone. Furthermore, E2-P22 binding was HA selective, as evidenced by negligible binding of the nanocages to non-HA materials including polystyrene, agarose, and polycaprolactone (PCL). Taken together these results establish a new mechanism for the directed coupling of nanocage drug delivery systems to a variety of HA-containing materials commonly used in diverse bone therapies. PMID:23312905

  20. Biological evaluation of porous aliphatic polyurethane/hydroxyapatite composite scaffolds for bone tissue engineering.

    PubMed

    Yang, Wanxun; Both, Sanne K; Zuo, Yi; Birgani, Zeinab Tahmasebi; Habibovic, Pamela; Li, Yubao; Jansen, John A; Yang, Fang

    2015-07-01

    Biomaterial scaffolds meant to function as supporting structures to osteogenic cells play a pivotal role in bone tissue engineering. Recently, we synthesized an aliphatic polyurethane (PU) scaffold via a foaming method using non-toxic components. Through this procedure a uniform interconnected porous structure was created. Furthermore, hydroxyapatite (HA) particles were introduced into this process to increase the bioactivity of the PU matrix. To evaluate the biological performances of these PU-based scaffolds, their influence on in vitro cellular behavior and in vivo bone forming capacity of the engineered cell-scaffold constructs was investigated in this study. A simulated body fluid test demonstrated that the incorporation of 40 wt % HA particles significantly promoted the biomineralization ability of the PU scaffolds. Enhanced in vitro proliferation and osteogenic differentiation of the seeded mesenchymal stem cells were also observed on the PU/HA composite. Next, the cell-scaffold constructs were implanted subcutaneously in a nude mice model. After 8 weeks, a considerable amount of vascularized bone tissue with initial marrow stroma development was generated in both PU and PU/HA40 scaffold. In conclusion, the PU/HA composite is a potential scaffold for bone regeneration applications. PMID:25370308

  1. Anisotropy of Longitudinal Wave Velocity and Hydroxyapatite Orientation in Bovine Cortical Bone

    NASA Astrophysics Data System (ADS)

    Yaoi, Yuichiro; Yamamoto, Kazufumi; Nakatsuji, Tomohiro; Yanagitani, Takahiko; Matsukawa, Mami; Yamazaki, Kaoru; Nagano, Akira

    2009-07-01

    The anisotropy of longitudinal wave velocity and the hydroxyapatite (HAp) crystallite orientation in bovine cortical bone were experimentally investigated in detail. A ring-shaped cortical bone sample was obtained from the midshaft of a 32-month-old bovine femur. Four cylindrical specimens (diameter 11 mm) were taken from the ring-shaped cortical bone specimen along the radial direction. Velocity was measured by a conventional ultrasonic pulse system, by rotating the specimen in the axial-tangential plane. The direction of the fastest wave velocity observed was inclined 5-10° from the axial direction. The HAp crystallite orientation in the specimen was also investigated by X-ray diffraction pole-figure analysis. We observed a strong intensive peak of (0002), indicating HAp crystallite c-axis alignment along the bone axis. However, the pole-figure also revealed a small tilt in the HAp crystallite orientation from the axial direction. Comparing the velocity and X-ray diffraction peak at each angle of the axial-tangential plane, a significant correlation between the velocity and the HAp orientation was observed.

  2. Engineering nanocages with polyglutamate domains for coupling to hydroxyapatite biomaterials and allograft bone

    PubMed Central

    Culpepper, Bonnie K.; Morris, David S.; Prevelige, Peter E.; Bellis, Susan L.

    2013-01-01

    Hydroxyapatite (HA) is the principal constituent of bone mineral, and synthetic HA is widely used as a biomaterial for bone repair. Previous work has shown that polyglutamate domains bind selectively to HA and that these domains can be utilized to couple bioactive peptides onto many different HA-containing materials. In the current study we have adapted this technology to engineer polyglutamate domains into cargo-loaded nanocage structures derived from the P22 bacteriophage. P22 nanocages have demonstrated significant potential as a drug delivery system due to their stability, large capacity for loading with a diversity of proteins and other types of cargo, and ability to resist degradation by proteases. Site-directed mutagenesis was used to modify the primary coding sequence of the P22 coat protein to incorporate glutamate-rich regions. Relative to wild-type P22, the polyglutamate-modified nanocages (E2-P22) exhibited increased binding to ceramic HA disks, particulate HA and allograft bone. Furthermore, E2-P22 binding was HA selective, as evidenced by negligible binding of the nanocages to non-HA materials including polystyrene, agarose, and polycaprolactone (PCL). Taken together these results establish a new mechanism for the directed coupling of nanocage drug delivery systems to a variety of HA-containing materials commonly used in diverse bone therapies. PMID:23312905

  3. Synthesis and characterization of a novel chitosan/montmorillonite/hydroxyapatite nanocomposite for bone tissue engineering.

    PubMed

    Katti, Kalpana S; Katti, Dinesh R; Dash, Rajalaxmi

    2008-09-01

    Recently, biopolymer-based nanocomposites have been replacing synthetic polymer composites for various biomedical applications. This is often because of the biocompatible and biodegradable behavior of natural polymers. Several studies have been reported pertaining to the synthesis and characterization of chitosan(chi)/montmorillonite(MMT) and chitosan (chi)/hydroxyapatite (HAP) for tissue engineering applications. In the present work, a biopolymer-based novel nanocomposite chitosan/montmorillonite (MMT)/hydroxyapatite (HAP) was developed for biomedical applications. The composite was prepared from chitosan, unmodified MMT and HAP precipitate in aqueous media. The properties of the composites were investigated using x-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM) and thermogravimetric analysis (TGA). Nanomechanical properties were measured using nanoindentation. Cell culture experiments were also conducted in order to ascertain the biocompatibility of the composite. The XRD results indicate that an intercalated structure was formed with an increase in d-spacing of montmorillonite. FTIR studies provide the evidence of molecular interaction among the three different constituents of the composite. AFM images show well-distributed nanoparticles in the chitosan matrix. The composites also exhibit a significant enhancement in nanomechanical property as compared to pure chitosan as well as the chi/HAP and chi/MMT composites. The TGA results indicate that an intercalated nanocomposite was formed with improved thermal properties even compared to chi/MMT composites. The results of cell culture experiments show that the composite is biocompatible and has a better cell proliferation rate compared to chi/HAP composites. This work represents the design of a novel clay-chitosan-hydroxyapatite composite with improved mechanical properties that has potential applications in bone tissue engineering. PMID:18765898

  4. Electrospun fibrous scaffold of hydroxyapatite/poly (ε-caprolactone) for bone regeneration.

    PubMed

    Li, Lingli; Li, Guang; Jiang, Jianming; Liu, Xiaona; Luo, Li; Nan, Kaihui

    2012-02-01

    Development of fibrous scaffold of hydroxyapatite/biopolymer nanocomposite offers great potential in the field of bone regeneration and tissue engineering. Hydroxyapatite (HA)/poly (ε-caprolactone) (PCL) fibrous scaffolds were successfully prepared by electrospinning dopes containing HA and PCL in this work. It was found that pre-treating HA with γ-glycioxypropyltrimethoxysilane (A-187) was effective in improving HA dispersion both in solutions and in a PCL matrix. Mechanical properties of the scaffolds were greatly enhanced by the filling of A187-HA. The bioactivity of PCL was remarkably improved by the addition of HA and A187-HA. Fibroblasts and osteoblasts were seeded on scaffolds to evaluate the effect of A-187 on biocompatibility of HA/PCL composites. Based on this study, good dispersion of HA in PCL matrix was granted by pretreatment of HA with A-187 and A187-HA/PCL fibrous scaffolds were obtained by electrospinning. These results demonstrated that the scaffolds may possess improved mechanical performance and good bioactivity due to A187-HA incorporation. PMID:22143907

  5. Cancellous bone healing around strontium-doped hydroxyapatite in osteoporotic rats previously treated with zoledronic acid.

    PubMed

    Li, Yunfeng; Shui, Xueping; Zhang, Li; Hu, Jing

    2016-04-01

    Bisphosphonates (BPs) are potent anti-osteoporotic agents. Strontium-doped hydroxyapatite (HA) (SrHA) has been reported to increase bone density and improve trabecular microarchitecture in osteoporotic animals. But information about the effect of SrHA on the surrounding bone tissue in osteoporotic animals previously on BPs treatment is limited. We hypothesize that SrHA will induce increased bone density in the vicinity of the material when compared to HA, even in osteoporotic animals previously treated with BPs. HA and 10%SrHA (HA with 10 mol % calcium substituted by strontium) implants were prepared and characterized by scanning electronic microscopy (SEM), X-ray photoemission spectroscopy (XPS), and X-ray diffraction (XRD). Osteoporotic animal model was established by bilateral ovariectomy. Twelve weeks later, all OVX rats accepted subcutaneous injection of zoledronic acid (ZOL) at the dose of 1.5 μg/kg weekly for another twelve weeks. Subsequently, rod-shaped HA and SrHA implants were inserted in the distal femur of the OVX animals previously treated with ZOL. Eight weeks after implantation, specimens were harvested for histological and micro-computed tomography (micro-CT) analysis. Compared to HA, 10%SrHA raised the percent bone volume by 32.7%, the mean trabecular thickness by 36.5%, the mean trabecular number by 34.3%, the mean connectivity density by 38.4%, while the mean trabecular separation showed no significant difference. 10%SrHA also increased the bone area density by 36.3% in histological analysis. Results from this study indicated that 10%SrHA increased bone density and improved trabecular microarchitecture around implants in osteoporotic animals previously treated with ZOL when compared to HA. PMID:25891947

  6. Development of a new carbon nanotube–alginate–hydroxyapatite tricomponent composite scaffold for application in bone tissue engineering

    PubMed Central

    Rajesh, Rajendiran; Dominic Ravichandran, Y

    2015-01-01

    In recent times, tricomponent scaffolds prepared from naturally occurring polysaccharides, hydroxyapatite, and reinforcing materials have been gaining increased attention in the field of bone tissue engineering. In the current work, a tricomponent scaffold with an oxidized multiwalled carbon nanotube (fMWCNT)–alginate–hydroxyapatite with the required porosity was prepared for the first time by a freeze-drying method and characterized using analytical techniques. The hydroxyapatite for the scaffold was isolated from chicken bones by thermal calcination at 800°C. The Fourier transform infrared spectra and X-ray diffraction data confirmed ionic interactions and formation of the fMWCNT–alginate–hydroxyapatite scaffold. Interconnected porosity with a pore size of 130–170 µm was evident from field emission scanning electron microscopy. The total porosity calculated using the liquid displacement method was found to be 93.85%. In vitro biocompatibility and cell proliferation on the scaffold was checked using an MG-63 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell attachment by Hoechst stain assay. In vitro studies showed better cell proliferation, cell differentiation, and cell attachment on the prepared scaffold. These results indicate that this scaffold could be a promising candidate for bone tissue engineering. PMID:26491303

  7. Bioinspired synthesis of hydroxyapatite nanocrystals in the presence of collagen and l-arginine: Candidates for bone regeneration.

    PubMed

    Brasinika, Despoina; Tsigkou, Olga; Tsetsekou, Athena; Missirlis, Yiannis F

    2016-04-01

    This work aims at the bioinspired synthesis of hydroxyapatite (HAp) crystals in the presence of both collagen and l-arginine, in an effort to obtain a homogeneous hybrid material, having a bone-like nanostructure. Collagen (Col) is the most commonly utilized protein in most species of life, while L-arginine (Arg) encourages cell attachment, proliferation, and differentiation on HAp surfaces. Transmission electron microscopy, X-ray diffraction and Fourier transform-infrared spectroscopy were used to analyze surface morphology and structure of nanocrystals obtained under different synthesis conditions. It was shown that collagen and arginine content affect HAp crystallization. Collagen has an inhibition effect since HAp crystal size is reduced with the increase of collagen content. The presence of arginine is crucial as a critical content exists (Ca(2+):Arg = 1:1) under which HAp nanocrystals coexist with brushite. Under the optimum synthesis conditions (HAp/Col weight ratio 70/30 and Ca(2+):Arg molar ratio 1:1) HAp nanoplates of a uniform size (around 10 × 10 nm) were obtained. The biocompatibility of this hybrid powder was assessed using human bone marrow derived mesenchymal stem cells (MSCs). Cell response in terms of MSC attachment (scanning electron microscopy) and viability/proliferation (Alamar Blue) demonstrated a noncytotoxic effect of the new material. PMID:25891542

  8. Hydroxyapatite ceramics from hydrothermally prepared powders

    SciTech Connect

    Lin, C.H.; Huang, C.W.; Chang, S.C.

    1994-12-31

    Hydroxyapatite (Ca{sub 5}(PO{sub 4}){sub 3}(OH)) is an effective material for artificial human bone production. Hydroxyapatite powders were hydrothermally produced in this work by reacting Ca(OH){sub 2} with Na{sub 3}PO{sub 4}{center_dot}12H{sub 2}O in an autoclave at various temperature and for various times. The particle size of hydroxyapatite was observed to be very fine, uniform, around 50 nm, as well as independent of reaction time. The hydroxyapatite powders were compacted and sintered at various temperatures for 2 hrs. The density, grain size, and hardness of the hydroxyapatite ceramics were measured and compared with those of the hydroxyapatite ceramics produced by the powders from the commercial source. The hydroxyapatite ceramics from the hydrothermal powders were found to have a higher density, smaller grain size, and higher hardness. After the hydroxyapatite ceramics were dipped in a simulated biological body liquid for 10 days, the density and hardness of the hydroxyapatite ceramics from the hydrothermal powders were less deteriorated than those of the hydroxyapatite ceramics from the commercial powder.

  9. Hydroxyapatite whisker reinforced 63s glass scaffolds for bone tissue engineering.

    PubMed

    Shuai, Cijun; Cao, Yiyuan; Gao, Chengde; Feng, Pei; Xiao, Tao; Peng, Shuping

    2015-01-01

    Bioactive glass (BG) is widely used for bone tissue engineering. However, poor mechanical properties are the major shortcomings. In the study, hydroxyapatite nanowhisker (HANw) was used as a reinforcement to improve the mechanical properties. 63s glass/HANw scaffolds were successfully fabricated by selective laser sintering (SLS). It was found that the optimal compressive strength and fracture toughness were achieved when 10 wt.% HANw was added. This led to 36% increase in compressive strength and 83% increase in fracture toughness, respectively, compared with pure 63s glass scaffolds. Different reinforcement mechanisms were analyzed based on the microstructure investigation. Whisker bridging and whisker pulling-out were efficient in absorbing crack propagating energy, resulting in the improvement of the mechanical properties. Moreover, bioactivity and biocompatibility of the scaffolds were evaluated in vitro. The results showed that composite scaffolds with 10 wt.% HANw exhibited good apatite-forming ability and cellular affinity. PMID:25821798

  10. Hydroxyapatite Whisker Reinforced 63s Glass Scaffolds for Bone Tissue Engineering

    PubMed Central

    Shuai, Cijun; Cao, Yiyuan; Gao, Chengde; Feng, Pei; Xiao, Tao; Peng, Shuping

    2015-01-01

    Bioactive glass (BG) is widely used for bone tissue engineering. However, poor mechanical properties are the major shortcomings. In the study, hydroxyapatite nanowhisker (HANw) was used as a reinforcement to improve the mechanical properties. 63s glass/HANw scaffolds were successfully fabricated by selective laser sintering (SLS). It was found that the optimal compressive strength and fracture toughness were achieved when 10 wt.% HANw was added. This led to 36% increase in compressive strength and 83% increase in fracture toughness, respectively, compared with pure 63s glass scaffolds. Different reinforcement mechanisms were analyzed based on the microstructure investigation. Whisker bridging and whisker pulling-out were efficient in absorbing crack propagating energy, resulting in the improvement of the mechanical properties. Moreover, bioactivity and biocompatibility of the scaffolds were evaluated in vitro. The results showed that composite scaffolds with 10 wt.% HANw exhibited good apatite-forming ability and cellular affinity. PMID:25821798

  11. Hydroxyapatite coating of cellulose sponge does not improve its osteogenic potency in rat bone.

    PubMed

    Ekholm, Erika; Tommila, Miretta; Forsback, Ari-Pekka; Märtson, Matis; Holmbom, Johanna; Aäritalo, Virpi; Finnberg, Christa; Kuusilehto, Asko; Salonen, Jukka; Yli-Urpo, Antti; Penttinen, Risto

    2005-09-01

    Regenerated cellulose sponges were coated biomimetically with hydroxyapatite to increase their osteogenic properties. Induction of apatite precipitation was carried out with bioactive glass in simulated body fluid (SBF) for 24 h and the final coating was carried out in 1.5 x concentrated SBF for 14 days. Biomimetically mineralized and non-mineralized sponges were then implanted into standard size femoral cortical defects of rats, and the invasion of bone into the implant was followed up to one year. The apatite coating did not improve the osteoconductive property of cellulose in this rat cortical defect model. In fact, it generated a strong and highly cellular inflammatory reaction and less osteoid tissue. The biomimetic implants contained more immunodetectable TGFbeta1 (a strong stimulator of fibroblast activity) than untreated implants, and also bound more TGFbeta1 in vitro, which could, at least in part, explain the fibrotic invasion of biomimetically mineralized sponges. PMID:16701833

  12. In vitro study on the degradation of lithium-doped hydroxyapatite for bone tissue engineering scaffold.

    PubMed

    Wang, Yaping; Yang, Xu; Gu, Zhipeng; Qin, Huanhuan; Li, Li; Liu, Jingwang; Yu, Xixun

    2016-09-01

    Li-doped hydroxyapatite (LiHA) which is prepared through introducing low dose of Li into hydroxyapatite (HA) has been increasingly studied as a bone tissue-engineered scaffold. The degradation properties play a crucial role in the success of long-term implantation of a bone tissue-engineered construct. Herein, the in vitro degradation behaviors of LiHA scaffolds via two approaches were investigated in this study: solution-mediated degradation and osteoblast-mediated degradation. In solution-mediated degradation, after being immersed in simulated body fluid (SBF) for some time, some characteristics of these scaffolds (such as release of ionized lithium and phosphate, pH change, mechanical properties, cytocompatibility and SEM surface characterization) were systematically tested. A similar procedure was also employed to research the degradation behaviors of LiHA scaffolds in osteoblast-mediated degradation. The results suggested that the degradation in SBF and degradation in culture medium with cell existed distinguishing mechanisms. LiHA scaffolds were degraded via a hydrolytic mechanism when they were soaked in SBF. Upon degradation, an apatite precipitation (layer) was formed on the surfaces of scaffolds. While a biological mechanism was presented for the degradation of scaffolds in cell-mediated degradation. Compared with pure HA, LiHA scaffolds had a better effect on the growth of osteoblast cells, meanwhile, the release amount of PO4(3-) in a degradation medium indicated that osteoblasts could accelerate the degradation of LiHA due to the more physiological activities of osteoblast. According to the results from compressive strength test, doping Li into HA could enhance the strength of HA. Moreover, the results from MTT assay and SEM observation showed that the degradation products of LiHA scaffolds were beneficial to the proliferation of osteoblasts. The results of this research can provide the theoretical basis for the clinical application of LiHA scaffolds

  13. Hydroxyapatite crystallinity does not affect the repair of critical size bone defects

    PubMed Central

    CONZ, Marcio Baltazar; GRANJEIRO, José Mauro; SOARES, Gloria de Almeida

    2011-01-01

    Objective The physicochemical properties of hydroxyapatite (HA) granules were observed to affect the biological behavior of graft materials. The aim of this work was to analyze the tissue response of two HA granules with different crystallinity and Ca/P ratio in vivo. Material and Methods The HA granules were produced in the Biomaterials Laboratory (COPPE/UFRJ). The testing materials were HA granules presenting a Ca/P molar ratio of 1.60 and 28% crystallinity (HA-1), and a Ca/P molar ratio of 1.67 and 70% crystallinity (HA-2). Both HAs were implanted into a critical-size calvaria rat defects. Results To note, in the control group, the bone defects were filled with blood clot only. Descriptive and histomorphometric analyses after 1, 3, and 6 months postoperatively showed mild inflammatory infiltrate, mainly comprising macrophage-like and multinucleated giant cells, and an increase in the volume density of the fibrous tissues (p<0.05), which was in contrast to the similar volume density of the newly formed bone and biomaterials in relation to the control group. Conclusion Thus, we concluded that HA-1 and HA-2 are biocompatible and non-degradable, and that crystallinity does not affect bone repair of critical size defects. PMID:21655775

  14. Aligned porous barium titanate/hydroxyapatite composites with high piezoelectric coefficients for bone tissue engineering.

    PubMed

    Zhang, Yan; Chen, Liangjian; Zeng, Jing; Zhou, Kechao; Zhang, Dou

    2014-06-01

    It was proposed that the piezoelectric effect played an important physiological role in bone growth, remodelling and fracture healing. An aligned porous piezoelectric composite scaffold was fabricated by freeze casting hydroxyapatite/barium titanate (HA/BT) suspensions. The highest compressive strength and lowest porosity of 14.5MPa and 57.4% with the best parallelism of the pore channels were achieved in the HA10/BT90 composite. HA30/BT70 and HA10/BT90 composites exhibited piezoelectric coefficient d33 of 1.2 and 2.8pC/N, respectively, both of which were higher than the piezoelectric coefficient of natural bone. Increase of the solid loading of the suspension and solidification velocity led to the improvement of piezoelectric coefficient d33. Meanwhile, double-templates resulted in the coexistence of lamellar pores and aligned macro-pores, exhibiting the ability to produce an oriented long-range ordered architecture. The manipulation flexibility of this method indicated the potential for customized needs in the application of bone substitute. An MTT assay indicated that the obtained scaffolds had no cytotoxic effects on L929 cells. PMID:24863210

  15. Functionalization of a Collagen-Hydroxyapatite Scaffold with Osteostatin to Facilitate Enhanced Bone Regeneration.

    PubMed

    Quinlan, Elaine; Thompson, Emmet M; Matsiko, Amos; O'Brien, Fergal J; López-Noriega, Adolfo

    2015-12-01

    Defects within bones caused by trauma and other pathological complications may often require the use of a range of therapeutics to facilitate tissue regeneration. A number of approaches have been widely utilized for the delivery of such therapeutics via physical encapsulation or chemical immobilization suggesting significant promise in the healing of bone defects. The study focuses on the chemical immobilization of osteostatin, a pentapeptide of the parathyroid hormone (PTHrP107-111), within a collagen-hydroxyapatite scaffold. The chemical attachment method via crosslinking supports as little as 4% release of the peptide from the scaffolds after 21 d whereas non-crosslinking leads to 100% of the peptide being released by as early as 4 d. In vitro characterization demonstrates that this cross-linking method of immobilization supports a pro-osteogenic effect on osteoblasts. Most importantly, when implanted in a critical-sized calvarial defect within a rat, these scaffolds promote significantly greater new bone volume and area compared to nonfunctionalized scaffolds (**p < 0.01) and an empty defect control (***p < 0.001). Collectively, this study suggests that such an approach of chemical immobilization offers greater spatiotemporal control over growth factors and can significantly modulate tissue regeneration. Such a system may be adopted for a range of different proteins and thus offers the potential for the treatment of various complex pathologies that require localized mediation of drug delivery. PMID:26414944

  16. Bone-repair properties of biodegradable hydroxyapatite nano-rod superstructures.

    PubMed

    D'Elía, Noelia L; Mathieu, Colleen; Hoemann, Caroline D; Laiuppa, Juan A; Santillán, Graciela E; Messina, Paula V

    2015-11-28

    Nano-hydroxyapatite (nano-HAp) materials show an analogous chemical composition to the biogenic mineral components of calcified tissues and depending on their topography they may mimic the specific arrangement of the crystals in bone. In this work, we have evaluated the potential of four synthesized nano-HAp superstructures for the in vitro conditions of bone-repair. Experiments are underway to investigate the effects of the material microstructure, surface roughness and hydrophilicity on their osseo-integration, osteo-conduction and osteo-induction abilities. Materials were tested in the presence of both, rat primary osteoblasts and rabbit mesenchymal stem cells. The following aspects are discussed: (i) cytotoxicity and material degradation; (ii) rat osteoblast spreading, proliferation and differentiation; and (iii) rabbit mesenchymal stem cell adhesion on nano-HAp and nano-HAp/collagen type I coatings. We effectively prepared a material based on biomimetic HAp nano-rods displaying the appropriate surface topography, hydrophilicity and degradation properties to induce the in vitro desired cellular responses for bone bonding and healing. Cells seeded on the selected material readily attached, proliferated and differentiated, as confirmed by cell viability, mitochondrial metabolic activity, alkaline phosphatase (ALP) activity and cytoskeletal integrity analysis by immunofluorescence localization of alpha-smooth muscle actin (α-SMA) protein. These results highlight the influence of material's surface characteristics to determine their tissue regeneration potential and their future use in engineering osteogenic scaffolds for orthopedic implants. PMID:26505580

  17. Brushite-based calcium phosphate cement with multichannel hydroxyapatite granule loading for improved bone regeneration.

    PubMed

    Sarkar, Swapan Kumar; Lee, Byung Yeol; Padalhin, Andrew Reyas; Sarker, Avik; Carpena, Nathaniel; Kim, Boram; Paul, Kallyanshish; Choi, Hwan Jun; Bae, Sang-Ho; Lee, Byong Taek

    2016-01-01

    In this work, we report brushite-based calcium phosphate cement (CPC) system to enhance the in vivo biodegradation and tissue in-growth by incorporation of micro-channeled hydroxyapatite (HAp) granule and silicon and sodium addition in calcium phosphate precursor powder. Sodium- and silicon-rich calcium phosphate powder with predominantly tri calcium phosphate (TCP) phase was synthesized by an inexpensive wet chemical route to react with mono calcium phosphate monohydrate (MCPM) for making the CPC. TCP nanopowder also served as a packing filler and moderator of the reaction kinetics of the setting mechanism. Strong sintered cylindrical HAp granules were prepared by fibrous monolithic (FM) process, which is 800 µm in diameter and have seven micro-channels. Acid sodium pyrophosphate and sodium citrate solution was used as the liquid component which acted as a homogenizer and setting time retarder. The granules accelerated the degradation of the brushite cement matrix as well as improved the bone tissue in-growth by permitting an easy access to the interior of the CPC through the micro-channels. The addition of micro-channeled granule in the CPC introduced porosity without sacrificing much of its compressive strength. In vivo investigation by creating a critical size defect in the femur head of a rabbit model for 1 and 2 months showed excellent bone in-growth through the micro-channels. The granules enhanced the implant degradation behavior and bone regeneration in the implanted area was significantly improved after two months of implantation. PMID:26333790

  18. Hydroxyapatite/polyurethane scaffold incorporated with drug-loaded ethyl cellulose microspheres for bone regeneration.

    PubMed

    Liu, Haohuai; Zhang, Li; Shi, Pujiang; Zou, Qin; Zuo, Yi; Li, Yubao

    2010-10-01

    The purpose of this study is to explore and develop biodegradable scaffold for bone regeneration or tissue engineering with the capacity of controlled drug delivery. Ceftazidime as a model drug was encapsulated in ethyl cellulose (EC) microspheres, which were subsequently incorporated in a hydroxyapatite/polyurethane (HA/PU) composite scaffold to generate an antibiotic drug delivery system. HA/PU scaffolds had an interconnected pore network with an average porosity of about 83%. The presence of microspheres in the composite scaffolds was confirmed by scanning electron microscopy. The drug-loaded EC microspheres were uniformly distributed in the HA/PU scaffold matrix and showed no significant effect on the pore structure of the scaffold. Incorporation of microspheres into scaffolds significantly reduced the initial burst release, and the system exhibited a sustained release of the model drug for up to 60 days. Moreover, the scaffold with drug-loaded microspheres was proved to be an effective drug delivery system with good cytocompatibility and antibacterial properties. The novel drug-loaded microsphere/scaffold composites developed in this study are promising to serve as vehicles for controlled drug delivery in bone regeneration or bone tissue engineering. PMID:20665683

  19. Increase of stability in external fracture fixation by hydroxyapatite-coated bone screws.

    PubMed

    Augat, P; Claes, L; Hanselmann, K F; Suger, G; Fleischmann, W

    1995-01-01

    A major problem in fracture treatment by external fixation is screw loosening, which often results in reduced stability and can lead to prolonged treatment. A load-carrying experiment was conducted to determine whether coating implants with bioactive hydroxyapatite (HA) increases screw stability. Twelve HA-coated ASIF screws with 3 different macroporosities were inserted in 12 sheep that had already been fitted with a 6-pin external fixator for the treatment of a tibial osteotomy. The same number of uncoated polished steel screws served as controls. Although initial stability was not different for HA-coated screws, average removal torque after a 9-week implantation period increased with increasing macroporosity of the HA coating (p < .002). Instability of some screws was accompanied by histologic findings of cartilagenous tissue and proliferation of periosteal callus. Near the threads in the tibial cortex and in the shaft area of the screw were seen large numbers of HA particles that had been sheared off during implantation as well as during screw removal because of high contact forces between the HA coating and bone. Particulate debris of HA particles as well as the release of small bone fragments during explanation is likely to be unavoidable since HA adherence to bone is greater than adherence to steel after several weeks of implantation. PMID:7640445

  20. A comparative biomechanical study of bone ingrowth in two porous hydroxyapatite bioceramics

    NASA Astrophysics Data System (ADS)

    Ren, Li-Mei; Todo, Mitsugu; Arahira, Takaaki; Yoshikawa, Hideki; Myoui, Akira

    2012-12-01

    Calcium phosphate-based bioceramics have been widely used as artificial bone substitute materials because of their superior biocompatibility and osteoconductivity. In the present study, mechanical properties changes of two hydroxyapatite (HA) ceramics induced by bone ingrowth were tested and evaluated in a rabbit model. Both materials (NEOBONE®, Apaceram-AX®) have highly interconnected pores with a porosity of 75-85%. The major structural difference between them lies in that Apaceram-AX® has micropores smaller than 10 micrometers in diameter, whereas NEOBONE® does not contain such micropores. Both materials were implanted into the femoral condyles of rabbits for the specified observation period (1, 5, 12, 24, and 48 weeks) and then evaluated by experimental approach in combination with finite element method (FEM). Results indicate that two porous bioceramics exhibit different degradability in vivo, and remarkably different variation of total stiffness, elastic modulus distribution, as well as strain energy density distribution calculated by FE simulation. These results demonstrate how the internal microstructures affect the progress of bone regeneration and mechanical properties with the duration of implantation, emphasizing the importance of biomaterial design tailored to various clinic applications. Additionally, this study showed a potential for applying the computational method to monitor the time-dependent biomechanical changes of implanted porous bioceramics.

  1. Effects of zinc-substituted nano-hydroxyapatite coatings on bone integration with implant surfaces*

    PubMed Central

    Zhao, Shi-fang; Dong, Wen-jing; Jiang, Qiao-hong; He, Fu-ming; Wang, Xiao-xiang; Yang, Guo-li

    2013-01-01

    Objective: The purpose of this study was to investigate the effects of a zinc-substituted nano-hydroxyapatite (Zn-HA) coating, applied by an electrochemical process, on implant osseointegraton in a rabbit model. Methods: A Zn-HA coating or an HA coating was deposited using an electrochemical process. Surface morphology was examined using field-emission scanning electron microscopy. The crystal structure and chemical composition of the coatings were examined using an X-ray diffractometer (XRD) and Fourier transform infrared spectroscopy (FTIR). A total of 78 implants were inserted into femurs and tibias of rabbits. After two, four, and eight weeks, femurs and tibias were retrieved and prepared for histomorphometric evaluation and removal torque (RTQ) tests. Results: Rod-like HA crystals appeared on both implant surfaces. The dimensions of the Zn-HA crystals seemed to be smaller than those of HA. XRD patterns showed that the peaks of both coatings matched well with standard HA patterns. FTIR spectra showed that both coatings consisted of HA crystals. The Zn-HA coating significantly improved the bone area within all threads after four and eight weeks (P<0.05), the bone to implant contact (BIC) at four weeks (P<0.05), and RTQ values after four and eight weeks (P<0.05). Conclusions: The study showed that an electrochemically deposited Zn-HA coating has potential for improving bone integration with an implant surface. PMID:23733429

  2. In vitro proliferation and osteogenic differentiation of human dental pulp stem cells in injectable thermo-sensitive chitosan/β-glycerophosphate/hydroxyapatite hydrogel.

    PubMed

    Chen, Yantian; Zhang, Fengli; Fu, Qiang; Liu, Yong; Wang, Zejian; Qi, Nianmin

    2016-09-01

    Injectable thermo-sensitive hydrogels have a potential application in bone tissue engineering for their sensitivities and minimal invasive properties. Human dental pulp stem cells have been considered a promising tool for tissue reconstruction. The objective of this study was to investigate the proliferation and osteogenic differentiation of dental pulp stem cells in injectable thermo-sensitive chitosan/β-glycerophosphate/hydroxyapatite hydrogel in vitro. The chitosan /β-glycerophosphate hydrogel and chitosan/β-glycerophosphate/hydroxyapatite hydrogel were prepared using the sol-gel method. The injectability of chitosan /β-glycerophosphate hydrogel and chitosan/β-glycerophosphate/hydroxyapatite hydrogel was measured using a commercial disposable syringe. Scanning electron microscopy was used to observe the inner structure of hydrogels. Then dental pulp stem cells were seeded in chitosan /β-glycerophosphate hydrogel and chitosan/β-glycerophosphate/hydroxyapatite hydrogel, respectively. The growth of dental pulp stem cells was periodically observed under an inverted microscope. The proliferation of dental pulp stem cells was detected by using an Alamar Blue kit, while cell apoptosis was determined by using a Live/Dead Viability/Cytotoxicity kit. The osteogenic differentiations of dental pulp stem cells in chitosan /β-glycerophosphate hydrogel and chitosan/β-glycerophosphate/hydroxyapatite hydrogel were evaluated by alkaline phosphatase activity assay and mRNA expression of osteogenesis gene for 21 days in osteogenic medium. The results indicated that there was no significant difference between chitosan /β-glycerophosphate hydrogel and chitosan/β-glycerophosphate/hydroxyapatite hydrogel in injectability. Cells within the chitosan/β-glycerophosphate/hydroxyapatite hydrogel displayed a typical adherent cell morphology and rapid proliferation with high cellular viability after 14 days of culture. Dental pulp stem cells seeded in chitosan/β-glycerophosphate/hydroxyapatite

  3. In vivo bone response to 3D periodic hydroxyapatite scaffolds assembled by direct ink writing.

    PubMed

    Simon, Joshua L; Michna, Sarah; Lewis, Jennifer A; Rekow, E Dianne; Thompson, Van P; Smay, James E; Yampolsky, Andrew; Parsons, J Russell; Ricci, John L

    2007-12-01

    The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures. PMID:17559109

  4. Vascular and micro-environmental influences on MSC-coral hydroxyapatite construct-based bone tissue engineering.

    PubMed

    Cai, Lei; Wang, Qian; Gu, Congmin; Wu, Jingguo; Wang, Jian; Kang, Ning; Hu, Jiewei; Xie, Fang; Yan, Li; Liu, Xia; Cao, Yilin; Xiao, Ran

    2011-11-01

    Bone tissue engineering (BTE) has been demonstrated an effective approach to generate bone tissue and repair bone defect in ectopic and orthotopic sites. The strategy of using a prevascularized tissue-engineered bone grafts (TEBG) fabricated ectopically to repair bone defects, which is called live bone graft surgery, has not been reported. And the quantitative advantages of vascularization and osteogenic environment in promoting engineered bone formation have not been defined yet. In the current study we generated a tissue engineered bone flap with a vascular pedicle of saphenous arteriovenous in which an organized vascular network was observed after 4 weeks implantation, and followed by a successful repaire of fibular defect in beagle dogs. Besides, after a 9 months long term observation of engineered bone formation in ectopic and orthotopic sites, four CHA (coral hydroxyapatite) scaffold groups were evaluated by CT (computed tomography) analysis. By the comparison of bone formation and scaffold degradation between different groups, the influences of vascularization and micro-environment on tissue engineered bone were quantitatively analyzed. The results showed that in the first 3 months vascularization improved engineered bone formation by 2 times of non-vascular group and bone defect micro-environment improved it by 3 times of ectopic group, and the CHA-scaffold degradation was accelerated as well. PMID:21855129

  5. Evaluation of Bone Healing on Sandblasted and Acid Etched Implants Coated with Nanocrystalline Hydroxyapatite: An In Vivo Study in Rabbit Femur

    PubMed Central

    Melin Svanborg, Lory; Meirelles, Luiz; Franke Stenport, Victoria; Currie, Fredrik; Andersson, Martin

    2014-01-01

    This study aimed at investigating if a coating of hydroxyapatite nanocrystals would enhance bone healing over time in trabecular bone. Sandblasted and acid etched titanium implants with and without a submicron thick coat of hydroxyapatite nanocrystals (nano-HA) were implanted in rabbit femur with healing times of 2, 4, and 9 weeks. Removal torque analyses and histological evaluations were performed. The torque analysis did not show any significant differences between the implants at any healing time. The control implant showed a tendency of more newly formed bone after 4 weeks of healing and significantly higher bone area values after 9 weeks of healing. According to the results from this present study, both control and nano-HA surfaces were biocompatible and osteoconductive. A submicron thick coating of hydroxyapatite nanocrystals deposited onto blasted and acid etched screw shaped titanium implants did not enhance bone healing, as compared to blasted and etched control implants when placed in trabecular bone. PMID:24723952

  6. The Behavior of Water in Collagen and Hydroxyapatite Sites of Cortical Bone: Fracture, Mechanical Wear, and Load Bearing Studies

    PubMed Central

    Gul-E-Noor, Farhana; Singh, Chandan; Papaioannou, Antonios; Sinha, Neeraj; Boutis, Gregory S.

    2015-01-01

    The mechanical properties of cortical bone, which is largely comprised of collagen, hydroxyapatite, and water, are known to hinge on hydration. Recently, the characteristics of water in bone have drawn attention as potential markers of bone quality. We report on the dynamics, diffusion, population, and exchange of water in cortical bone by NMR relaxation and diffusion methodologies. Relaxation measurements over timescales ranging from 0.001 to 4.2 s reveal two distinguishable water environments. Systematic exposure to ethylenediaminetetraacetic acid or collagenase reveals one peak in our 2D relaxation map belonging to water present in the hydroxyapatite rich environment, and a second peak with shorter relaxation times arising from a collagen rich site. Diffusion-T2 measurements allowed for direct measurement of the diffusion coefficient of water in all observable reservoirs. Further, deuterium relaxation methods were applied to study cortical bone under an applied force, following mechanical wear or fracture. The tumbling correlation times of water reduce in all three cases, indicating that water dynamics may be used as a probe of bone quality. Lastly, changes in the relative populations and correlation times of water in bone under an applied force suggest that load bearing occurs largely in the collagen rich environment and is reversible. PMID:26659838

  7. In vitro and in vivo bioactivity assessment of a polylactic acid/hydroxyapatite composite for bone regeneration

    PubMed Central

    Danoux, Charlène B; Barbieri, Davide; Yuan, Huipin; de Bruijn, Joost D; van Blitterswijk, Clemens A; Habibovic, Pamela

    2014-01-01

    Synthetic bone graft substitutes based on composites consisting of a polymer and a calcium-phosphate (CaP) ceramic are developed with the aim to satisfy both mechanical and bioactivity requirements for successful bone regeneration. In the present study, we have employed extrusion to produce a composite consisting of 50 wt.% poly(D,L-lactic acid) (PLA) and 50 wt.% nano-sized hydroxyapatite (HA) powder, achieving homogeneous distribution of the ceramic within the polymeric phase. In vitro, in both a simulated physiological saline (SPS) and a simulated body fluid (SBF), a greater weight loss was observed for PLA/HA than for PLA particles upon 12-week immersion. Furthermore, in SPS, a continuous release of calcium and phosphate from the composite was measured, whereas in SBF, decrease of the amount of the two ions in the solution was observed both for PLA and PLA/HA accompanied with the formation of a CaP layer on the surface. In vitro characterization of the composite bioactivity was performed by culturing human mesenchymal stromal cells (hMSCs) and assessing proliferation and osteogenic differentiation, with PLA as a control. Both PLA/HA composite and PLA control were shown to support hMSCs proliferation over a period of two weeks. In addition, the composite significantly enhanced alkaline phosphatase (ALP) activity of hMSCs in osteogenic medium as compared with the polymer control. A novel implant design was employed to develop implants from dense, extruded materials, suitable for testing osteoinductivity in vivo. In a preliminary study in dogs, PLA/HA composite implants induced heterotopic bone formation upon 12-week intramuscular implantation in all animals, in contrast to PLA control, which was not osteoinductive. Unlike in vitro, a more pronounced degradation of PLA was observed in vivo as compared with PLA/HA composite. PMID:24441389

  8. In vitro and in vivo bioactivity assessment of a polylactic acid/hydroxyapatite composite for bone regeneration.

    PubMed

    Danoux, Charlène B; Barbieri, Davide; Yuan, Huipin; de Bruijn, Joost D; van Blitterswijk, Clemens A; Habibovic, Pamela

    2014-01-01

    Synthetic bone graft substitutes based on composites consisting of a polymer and a calcium-phosphate (CaP) ceramic are developed with the aim to satisfy both mechanical and bioactivity requirements for successful bone regeneration. In the present study, we have employed extrusion to produce a composite consisting of 50 wt.% poly(D,L-lactic acid) (PLA) and 50 wt.% nano-sized hydroxyapatite (HA) powder, achieving homogeneous distribution of the ceramic within the polymeric phase. In vitro, in both a simulated physiological saline (SPS) and a simulated body fluid (SBF), a greater weight loss was observed for PLA/HA than for PLA particles upon 12-week immersion. Furthermore, in SPS, a continuous release of calcium and phosphate from the composite was measured, whereas in SBF, decrease of the amount of the two ions in the solution was observed both for PLA and PLA/HA accompanied with the formation of a CaP layer on the surface. In vitro characterization of the composite bioactivity was performed by culturing human mesenchymal stromal cells (hMSCs) and assessing proliferation and osteogenic differentiation, with PLA as a control. Both PLA/HA composite and PLA control were shown to support hMSCs proliferation over a period of two weeks. In addition, the composite significantly enhanced alkaline phosphatase (ALP) activity of hMSCs in osteogenic medium as compared with the polymer control. A novel implant design was employed to develop implants from dense, extruded materials, suitable for testing osteoinductivity in vivo. In a preliminary study in dogs, PLA/HA composite implants induced heterotopic bone formation upon 12-week intramuscular implantation in all animals, in contrast to PLA control, which was not osteoinductive. Unlike in vitro, a more pronounced degradation of PLA was observed in vivo as compared with PLA/HA composite. PMID:24441389

  9. Hydroxyapatite Formation on a Novel Dental Cement in Human Saliva

    PubMed Central

    Engstrand, Johanna; Unosson, Erik; Engqvist, Håkan

    2012-01-01

    Dental materials have to meet high standards regarding mechanical strength and handling properties. There is however only a limited amount of research that has been devoted to natural formation of hydroxyapatite (HA) in contact with the materials. The objective of the current investigation was to study the surface reactions occurring in human salvia on a novel dental cement. Ceramir Crown & Bridge, a bioceramic luting agent intended for permanent cementation of conventional oral prosthetics, was evaluated by immersing discs made from the cement in human saliva and phosphate buffered saline (PBS) for seven days, after which they were dried and analyzed. The analytical methods used in order to verify HA formation on the surface were grazing incidence X-ray diffraction (GI-XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS). All results showed that HA was formed on the surfaces of samples stored in saliva as well as on samples stored in PBS. The possibility of a dental luting cement to promote natural formation of HA at the tooth interface increases the stability and durability of the system and could help prevent secondary caries. PMID:23056955

  10. Morphology of the osteonal cement line in human bone

    SciTech Connect

    Schaffler, M.B.; Burr, D.B.; Frederickson, R.G.

    1987-03-01

    While current consensus suggests the absence of collagen in osteonal cement lines, the extent of cement line mineralization and the nature of the ground substance within the cement line are unclear. Samples of human radius were examined by using scanning electron microscopy, electron microprobe, and histochemical techniques. X-ray intensities were used to compare the amount of calcium, phosphorus, and sulfur in cement lines with amounts in surrounding lamellar bone. The results indicate that cement lines contain significantly less calcium and phosphorus, but significantly more sulfur, than surrounding bone matrix. The Ca/P ratio of cement lines was significantly greater than that of lamellar bone, suggesting that the mineral in cement lines may not be in the form of mature hydroxyapatite. No selective staining of the cement lines could be demonstrated by using periodic acid-Schiff, Sudan black B, or alcian blue critical electrolyte concentration techniques.

  11. Phenotype expression of human bone cells cultured on implant substrates.

    PubMed

    Locci, P; Becchetti, E; Pugliese, M; Rossi, L; Belcastro, S; Calvitti, M; Pietrarelli, G; Staffolani, N

    1997-09-01

    Bone cells derived from the human jaw were cultured on titanium, titanium coated with hydroxyapatite (THA) or with plasma spray (TPS) to study the behaviour of the cells anchored to implant substrates. Bone cells were cultured in MEM with the addition of [3H]-thymidine to evaluate cellular proliferation, and [3H]-glucosamine to evaluate GAG synthesis and accumulation in the extra-cellular matrix (ECM). Moreover, to study the degradation of GAG bone cells were cultured in the presence of NH4Cl, an amine known to inhibit lysosomal activity. Our results show that TPS is the substrate that favours both cellular proliferation and the accumulation of GAG in the ECM. PMID:9377794

  12. Development of porous polyurethane/strontium-substituted hydroxyapatite composites for bone regeneration.

    PubMed

    Sariibrahimoglu, Kemal; Yang, Wanxun; Leeuwenburgh, Sander C G; Yang, Fang; Wolke, Joop G C; Zuo, Yi; Li, Yubao; Jansen, John A

    2015-06-01

    Polyurethane (PU) has been widely used for the biomedical applications but its potential for bone regeneration is limited due to its lack of osteoconductive properties. Strontium substituted hydroxyapatite (SrHA) particles, on the other hand, are known to exhibit a positive effect on bone formation. Therefore, the aim of this study was to (i) develop porous polyurethane scaffolds containing strontium SrHA nanoparticles (PU/SrHA) and (ii) compare their in vitro biological performance for applications in bone regeneration to PU scaffolds. SrHA and HA was synthesized using a conventional wet-chemical neutralization reaction at temperatures of 25, 50, and 80°C. Chemical analysis was performed by inductively coupled plasma-optical emission spectrometry. Synthesizing temperatures at 25 and at 50°C were selected for the composite preparation (abbreviated as HA-25, SrHA-25, HA-50, and SrHA-50, respectively). PU was synthesized from isophorone diisocyanate, polytetramethylene ether glycol, and 1,4-butanediol. Composite scaffolds were prepared by addition of HA or SrHA nanoparticles into PU scaffolds during polymer preparation. The results showed that the Sr content in HA nanoparticles increased with increasing synthesis temperature. The addition of nanoparticles decreased the elongation-at-break and tensile strength, but significantly increased the surface wettability of the PU scaffolds. In vitro degradation tests demonstrated that release of cations was significantly higher from PU/SrHA-50 composite scaffolds. Cell culture tests indicated that PU composites containing either HA or SrHA nanoparticles increased proliferation of bone marrow stem cells as compared to plain PU scaffolds, whereas osteogenic differentiation was not affected by the incorporation of HA nanoparticles irrespective of the incorporation of Sr. PMID:25203691

  13. Microsphere-based scaffolds encapsulating tricalcium phosphate and hydroxyapatite for bone regeneration.

    PubMed

    Gupta, Vineet; Lyne, Dina V; Barragan, Marilyn; Berkland, Cory J; Detamore, Michael S

    2016-07-01

    Bioceramic mixtures of tricalcium phosphate (TCP) and hydroxyapatite (HAp) are widely used for bone regeneration because of their excellent cytocompatibility, osteoconduction, and osteoinduction. Therefore, we hypothesized that incorporation of a mixture of TCP and HAp in microsphere-based scaffolds would enhance osteogenesis of rat bone marrow stromal cells (rBMSCs) compared to a positive control of scaffolds with encapsulated bone-morphogenic protein-2 (BMP-2). Poly(D,L-lactic-co-glycolic acid) (PLGA) microsphere-based scaffolds encapsulating TCP and HAp mixtures in two different ratios (7:3 and 1:1) were fabricated with the same net ceramic content (30 wt%) to evaluate how incorporation of these ceramic mixtures would affect the osteogenesis in rBMSCs. Encapsulation of TCP/HAp mixtures impacted microsphere morphologies and the compressive moduli of the scaffolds. Additionally, TCP/HAp mixtures enhanced the end-point secretion of extracellular matrix components relevant to bone tissue compared to the "blank" (PLGA-only) microsphere-based scaffolds as evidenced by the biochemical, gene expression, histology, and immunohistochemical characterization. Moreover, the TCP/HAp mixture groups even surpassed the BMP-2 positive control group in some instances in terms of matrix synthesis and gene expression. Lastly, gene expression data suggested that the rBMSCs responded differently to different TCP/HAp ratios presented to them. Altogether, it can be concluded that TCP/HAp mixtures stimulated the differentiation of rBMSCs toward an osteoblastic phenotype, and therefore may be beneficial in gradient microsphere-based scaffolds for osteochondral regeneration. PMID:27272903

  14. Similar healthy osteoclast and osteoblast activity on nanocrystalline hydroxyapatite and nanoparticles of tri-calcium phosphate compared to natural bone

    PubMed Central

    MacMillan, Adam K; Lamberti, Francis V; Moulton, Julia N; Geilich, Benjamin M; Webster, Thomas J

    2014-01-01

    While there have been numerous studies to determine osteoblast (bone forming cell) functions on nanocrystalline compared to micron crystalline ceramics, there have been few studies which have examined osteoclast activity (including tartrate-resistant acid phosphatase, formation of resorption pits, size of resorption pits, and receptor activator of nuclear factor κB [RANK]). This is despite the fact that osteoclasts are an important part of maintaining healthy bone since they resorb bone during the bone remodeling process. Moreover, while it is now well documented that bone formation is enhanced on nanoceramics compared to micron ceramics, some have pondered whether osteoblast functions (such as osteoprotegerin and RANK ligand [RANKL]) are normal (ie, non-diseased) on such materials compared to natural bone. For these reasons, the objective of the present in vitro study was to determine various functions of osteoclasts and osteoblasts on nanocrystalline and micron crystalline hydroxyapatite as well as tri-calcium phosphate materials and compare such results to cortical and cancellous bone. Results showed for the first time similar osteoclast activity (including tartrate-resistant acid phosphatase, formation of resorption pits, size of resorption pits, and RANK) and osteoblast activity (osteoprotegerin and RANKL) on nanocrystalline hydroxyapatite compared to natural bone, whereas osteoclast and osteoblast functions on micron crystalline versions of these ceramics were much different than natural bone. In this manner, this study provides additional evidence that nanocrystalline calcium phosphates can serve as suitable synthetic analogs to natural bone to improve numerous orthopedic applications. It also provides the first data of healthy osteoclast and osteoblast functions on nanocrystalline calcium phosphates compared to natural bone. PMID:25506216

  15. Effect of bioactive borate glass microstructure on bone regeneration, angiogenesis, and hydroxyapatite conversion in a rat calvarial defect model.

    PubMed

    Bi, Lianxiang; Rahaman, Mohamed N; Day, Delbert E; Brown, Zackary; Samujh, Christopher; Liu, Xin; Mohammadkhah, Ali; Dusevich, Vladimir; Eick, J David; Bonewald, Lynda F

    2013-08-01

    Borate bioactive glasses are biocompatible and enhance new bone formation, but the effect of their microstructure on bone regeneration has received little attention. In this study scaffolds of borate bioactive glass (1393B3) with three different microstructures (trabecular, fibrous, and oriented) were compared for their capacity to regenerate bone in a rat calvarial defect model. 12weeks post-implantation the amount of new bone, mineralization, and blood vessel area in the scaffolds were evaluated using histomorphometric analysis and scanning electron microscopy. The amount of new bone formed was 33%, 23%, and 15%, respectively, of the total defect area for the trabecular, oriented, and fibrous microstructures. In comparison, the percent new bone formed in implants composed of silicate 45S5 bioactive glass particles (250-300μm) was 19%. Doping the borate glass with copper (0.4 wt.% CuO) had little effect on bone regeneration in the trabecular and oriented scaffolds, but significantly enhanced bone regeneration in the fibrous scaffolds (from 15 to 33%). The scaffolds were completely converted to hydroxyapatite within the 12week implantation. The amount of hydroxyapatite formed, 22%, 35%, and 48%, respectively, for the trabecular, oriented, and fibrous scaffolds, increased with increasing volume fraction of glass in the as-fabricated scaffold. Blood vessels infiltrated into all the scaffolds, but the trabecular scaffolds had a higher average blood vessel area compared with the oriented and fibrous scaffolds. While all three scaffold microstructures were effective in supporting bone regeneration, the trabecular scaffolds supported more bone formation and may be more promising in bone repair. PMID:23643606

  16. Composite Scaffolds Containing Silk Fibroin, Gelatin, and Hydroxyapatite for Bone Tissue Regeneration and 3D Cell Culturing

    PubMed Central

    Moisenovich, M. M.; Arkhipova, A. Yu.; Orlova, A. A.; Drutskaya, M. S; Volkova, S. V.; Zacharov, S. E.; Agapov, I. I.; Kirpichnikov, M. P.

    2014-01-01

    Three-dimensional (3D) silk fibroin scaffolds were modified with one of the major bone tissue derivatives (nano-hydroxyapatite) and/or a collagen derivative (gelatin). Adhesion and proliferation of mouse embryonic fibroblasts (MEF) within the scaffold were increased after modification with either nano-hydroxyapatite or gelatin. However, a significant increase in MEF adhesion and proliferation was observed when both additives were introduced into the scaffold. Such modified composite scaffolds provide a new and better platform to study wound healing, bone and other tissue regeneration, as well as artificial organ bioengineering. This system can further be applied to establish experimental models to study cell-substrate interactions, cell migration and other complex processes, which may be difficult to address using the conventional two-dimensional culture systems. PMID:24772332

  17. Preparation of collagen/hydroxyapatite/alendronate hybrid hydrogels as potential scaffolds for bone regeneration.

    PubMed

    Ma, Xin; He, Zhiwei; Han, Fengxuan; Zhong, Zhiyuan; Chen, Liang; Li, Bin

    2016-07-01

    Development of biomimetic scaffolds represents a promising direction in bone tissue engineering. In this study, we designed a two-step process to prepare a type of biomimetic hybrid hydrogels that were composed of collagen, hydroxyapatite (HAP) and alendronate (ALN), an anti-osteoporosis drug. First, water-soluble ALN-conjugated HAP (HAP-ALN) containing 4.0wt.% of ALN was synthesized by treating HAP particles with ALN. Hydrogels were then formed from HAP-ALN conjugate and collagen under physiological conditions using genipin (GNP) as the crosslinker. Depending on the ALN/collagen molar ratio and GNP concentration, the gelation time of hydrogels ranged from 5 to 37min. Notably, these hybrid hydrogels exhibited markedly improved mechanical property (storage modulus G'=38-187kPa), higher gel contents, and lower swelling ratios compared to the hydrogels prepared from collagen alone under similar conditions. Moreover, they showed tunable degradation behaviors against collagenase. The collagen/HAP-ALN hybrid hydrogels supported the adhesion and growth of murine MC3T3-E1 osteoblastic cells well. Such tough yet enzymatically degradable hybrid hydrogels hold potential as scaffolds for bone tissue engineering. PMID:26998869

  18. Carbon nanotubes in nanocomposites and hybrids with hydroxyapatite for bone replacements.

    PubMed

    Shin, Ueon Sang; Yoon, Il-Kyu; Lee, Gil-Su; Jang, Won-Cheoul; Knowles, Jonathan C; Kim, Hae-Won

    2011-01-01

    Hydroxyapatite (HA), as a bone mineral component, has been an attractive bioceramic for the reconstruction of hard tissues. However, its poor mechanical properties, including low fracture toughness and tensile strength, have been a significant challenge to the application of HA for the replacement of load-bearing and/or large bone defects. Among materials studied to reinforce HA, carbon nanotubes (CNTs: single-walled or multiwalled) have recently gained significant attention because of their unprecedented mechanical properties (high strength and toughness) and physicochemical properties (high surface area, electrical and thermal conductivity, and low weight). Here, we review recent studies of the organization of HA-CNTs at the nanoscale, with a particular emphasis on the functionalization of CNTs and their dispersion within an HA matrix and induction of HA mineralization. The organization of CNTs and HA implemented at the nanoscale can further be developed in the form of coatings, nanocomposites, and hybrid powders to enable potential applications in hard tissue reconstruction. PMID:21776341

  19. Adverse Biological Effect of TiO₂ and Hydroxyapatite Nanoparticles Used in Bone Repair and Replacement.

    PubMed

    Wang, Jiangxue; Wang, Liting; Fan, Yubo

    2016-01-01

    The adverse biological effect of nanoparticles is an unavoidable scientific problem because of their small size and high surface activity. In this review, we focus on nano-hydroxyapatite and TiO₂ nanoparticles (NPs) to clarify the potential systemic toxicological effect and cytotoxic response of wear nanoparticles because they are attractive materials for bone implants and are widely investigated to promote the repair and reconstruction of bone. The wear nanoparticles would be prone to binding with proteins to form protein-particle complexes, to interacting with visible components in the blood including erythrocytes, leukocytes, and platelets, and to being phagocytosed by macrophages or fibroblasts to deposit in the local tissue, leading to the formation of fibrous local pseudocapsules. These particles would also be translocated to and disseminated into the main organs such as the lung, liver and spleen via blood circulation. The inflammatory response, oxidative stress, and signaling pathway are elaborated to analyze the potential toxicological mechanism. Inhibition of the oxidative stress response and signaling transduction may be a new therapeutic strategy for wear debris-mediated osteolysis. Developing biomimetic materials with better biocompatibility is our goal for orthopedic implants. PMID:27231896

  20. Carbon Nanotubes in Nanocomposites and Hybrids with Hydroxyapatite for Bone Replacements

    PubMed Central

    Shin, Ueon Sang; Yoon, Il-Kyu; Lee, Gil-Su; Jang, Won-Cheoul; Knowles, Jonathan C.; Kim, Hae-Won

    2011-01-01

    Hydroxyapatite (HA), as a bone mineral component, has been an attractive bioceramic for the reconstruction of hard tissues. However, its poor mechanical properties, including low fracture toughness and tensile strength, have been a significant challenge to the application of HA for the replacement of load-bearing and/or large bone defects. Among materials studied to reinforce HA, carbon nanotubes (CNTs: single-walled or multiwalled) have recently gained significant attention because of their unprecedented mechanical properties (high strength and toughness) and physicochemical properties (high surface area, electrical and thermal conductivity, and low weight). Here, we review recent studies of the organization of HA-CNTs at the nanoscale, with a particular emphasis on the functionalization of CNTs and their dispersion within an HA matrix and induction of HA mineralization. The organization of CNTs and HA implemented at the nanoscale can further be developed in the form of coatings, nanocomposites, and hybrid powders to enable potential applications in hard tissue reconstruction. PMID:21776341

  1. PCL-coated hydroxyapatite scaffold derived from cuttlefish bone: in vitro cell culture studies.

    PubMed

    Milovac, Dajana; Gamboa-Martínez, Tatiana C; Ivankovic, Marica; Gallego Ferrer, Gloria; Ivankovic, Hrvoje

    2014-09-01

    In the present study, we examined the potential of using highly porous poly(ε-caprolactone) (PCL)-coated hydroxyapatite (HAp) scaffold derived from cuttlefish bone for bone tissue engineering applications. The cell culture studies were performed in vitro with preosteoblastic MC3T3-E1 cells in static culture conditions. Comparisons were made with uncoated HAp scaffold. The attachment and spreading of preosteoblasts on scaffolds were observed by Live/Dead staining Kit. The cells grown on the HAp/PCL composite scaffold exhibited greater spreading than cells grown on the HAp scaffold. DNA quantification and scanning electron microscopy (SEM) confirmed a good proliferation of cells on the scaffolds. DNA content on the HAp/PCL scaffold was significantly higher compared to porous HAp scaffolds. The amount of collagen synthesis was determined using a hydroxyproline assay. The osteoblastic differentiation of the cells was evaluated by determining alkaline phosphatase (ALP) activity and collagen type I secretion. Furthermore, cell spreading and cell proliferation within scaffolds were observed using a fluorescence microscope. PMID:25063118

  2. A novel squid pen chitosan/hydroxyapatite/β-tricalcium phosphate composite for bone tissue engineering.

    PubMed

    Shavandi, Amin; Bekhit, Alaa El-Din A; Sun, Zhifa; Ali, Azam; Gould, Maree

    2015-10-01

    Squid pen chitosan was used in the fabrication of biocomposite scaffolds for bone tissue engineering. Hydroxyapatite (HA) and beta-tricalcium phosphate (β-TCP) obtained from waste mussel shells were used as the calcium phosphate source. The composite was prepared using 2.5% tripolyphosphate (TPP) and 1% glycerol as a cross-linker and plasticizer, respectively. The weight percent (wt.%) ratios of the ceramic components in the composite were 20/10/70, 30/20/50 and 40/30/30 (HA/β-TCP/Chi). The biodegradation rate and structural properties of the scaffolds were investigated. Scanning electron microscopy (SEM) and microCT(μCT) results indicated that the composites have a well defined lamellar structure with an average pore size of 200 μm. The porosity of the composites decreased from 88 to 56% by increasing the ratio of HA/β-TCP from 30 to 70%. After 28 days of incubation in a physiological solution, the scaffolds were degraded by approximately 30%. In vitro investigations showed that the composites were cytocompatible and supported the growth of L929 and Saos-2 cells. The obtained data suggests that the squid pen chitosan composites are potential candidates for bone regeneration. PMID:26117768

  3. Characterization of natural nanostructured hydroxyapatite obtained from the bones of Brazilian river fish

    NASA Astrophysics Data System (ADS)

    Coelho, T. M.; Nogueira, E. S.; Steimacher, A.; Medina, A. N.; Weinand, W. R.; Lima, W. M.; Baesso, M. L.; Bento, A. C.

    2006-11-01

    In this study, the characterization of the optical properties of natural hydroxyapatite (HAp) [Ca10(PO4)6(OH)2] is discussed. In the first stage of the experiment, natural HAp was processed from the bones of Brazilian river fish such as pintado (Pseudoplatystoma corruscans), jaú (Paulicea lutkeni), and cachara (Pseudoplatystoma fasciatum). The bones were calcined at 900°C for different amounts of time (4-12h) and reduced to powder using two different milling times (2 and 4h) in a high-energy ball mill, in order to determine the best procedure for obtaining natural nanostructured HAp powder for the study. In the second stage, material calcined for 8h was milled for 2, 4, 8, and 16h. The techniques of photoacoustic spectroscopy, scanning electron microscopy, and flame atomic absorption spectrometry were applied to characterize these samples. The O-H stretching shown by photoacoustic spectroscopy was correlated to the HAp nanostructure. Structural analysis indicated a Ca /P ratio close to 1.67 (˜1.64±0.04) and the presence of morphology and particle phase structure in the nanostructured HAp powder.

  4. In vitro deposition of hydroxyapatite on cortical bone collagen stimulated by deformation-induced piezoelectricity.

    PubMed

    Noris-Suárez, Karem; Lira-Olivares, Joaquin; Ferreira, Ana Marina; Feijoo, José Luis; Suárez, Nery; Hernández, Maria C; Barrios, Esteban

    2007-03-01

    In the present work, we have studied the effect of the piezoelectricity of elastically deformed cortical bone collagen on surface using a biomimetic approach. The mineralization process induced as a consequence of the piezoelectricity effect was evaluated using scanning electron microscopy (SEM), thermally stimulated depolarization current (TSDC), and differential scanning calorimetry (DSC). SEM micrographs showed that mineralization occurred predominantly over the compressed side of bone collagen, due to the effect of piezoelectricity, when the sample was immersed in the simulated body fluid (SBF) in a cell-free system. The TSDC method was used to examine the complex collagen dielectric response. The dielectric spectra of deformed and undeformed collagen samples with different hydration levels were compared and correlated with the mineralization process followed by SEM. The dielectric measurements showed that the mineralization induced significant changes in the dielectric spectra of the deformed sample. DSC and TSDC results demonstrated a reduction of the collagen glass transition as the mineralization process advanced. The combined use of SEM, TSDC, and DSC showed that, even without osteoblasts present, the piezoelectric dipoles produced by deformed collagen can produce the precipitation of hydroxyapatite by electrochemical means, without a catalytic converter as occurs in classical biomimetic deposition. PMID:17261065

  5. Preparation and characterization of (PCL-crosslinked-PEG)/hydroxyapatite as bone tissue engineering scaffolds.

    PubMed

    Koupaei, Narjes; Karkhaneh, Akbar; Daliri Joupari, Morteza

    2015-12-01

    In this study, interconnected porous bioactive scaffolds were synthesized for bone tissue engineering. At the first step, poly( ɛ-caprolactone) (PCL) diols were diacrylated with acryloyl chloride. Then, the scaffolds were synthesized by radical crosslinking reaction of PCL and poly(ethyleneglycol) (PEG) diacrylates in the presence of hydroxyapatite (HA) particles. Morphological, swelling, thermal, and mechanical characteristics as well as degradability of the scaffolds were investigated. Results showed that increasing the ratio of PEG to PCL led to significant increase of swelling ratio and degradation rate, and decrease of crystallinity and compressive modulus of the networks, respectively. It was found that the incorporation of HA particles with the polymer matrices resulted in an augmented crystallinity, a decreased swelling ratio, and also a significantly increased compressive modulus of the networks. Cytocompatability and osteoconductivity of the scaffolds were assessed by MTT and alkaline phosphatase (ALP) assays, respectively. The results confirmed the cytocompatible nature of PCL/PEG/HA scaffolds with no toxicity. MG-63 cells attached and spread on the pore walls offered by the scaffolds. PCL/PEG/HA scaffolds compared with PCL/PEG ones showed higher ALP activity. Thus, the results indicated that the PCL/PEG/HA scaffolds have the potential of being used as promising substrates in bone tissue engineering. PMID:26015080

  6. Hydroxyapatite nanoparticle injectable hydrogel scaffold to support osteogenic differentiation of human mesenchymal stem cells.

    PubMed

    Thorpe, A A; Creasey, S; Sammon, C; Le Maitre, C L

    2016-01-01

    Bone loss associated with degenerative disease and trauma is a clinical problem increasing with the aging population. Thus, effective bone augmentation strategies are required; however, many have the disadvantages that they require invasive surgery and often the addition of expensive growth factors to induce osteoblast differentiation. Here, we investigated a LaponiteÒ crosslinked, pNIPAM-DMAc copolymer (L-pNIPAM-co-DMAc) hydrogel with hydroxyapatite nanoparticles (HAPna), which can be maintained as a liquid ex vivo, injected via narrow-gauge needle into affected bone, followed by in situ gelation to deliver and induce osteogenic differentiation of human mesenchymal stem cells (hMSC). L-pNIPAM-co-DMAc hydrogels were synthesised and HAPna added post polymerisation. Commercial hMSCs from one donor (Lonza) were incorporated in liquid hydrogel, the mixture solidified and cultured for up to 6 weeks. Viability of hMSCs was maintained within hydrogel constructs containing 0.5 mg/mL HAPna. SEM analysis demonstrated matrix deposition in cellular hydrogels which were absent in acellular controls. A significant increase in storage modulus (G') was observed in cellular hydrogels with 0.5 mg/mL HAPna. Semi-quantitative immunohistochemistry and histological analysis demonstrated that bone differentiation markers and collagen deposition was induced within 48 h, with increased calcium deposition with time. The thermally triggered hydrogel system, described here, was sufficient without the need of additional growth factors or osteogenic media to induce osteogenic differentiation of commercial hMSCs. Preliminary data presented here will be expanded on multiple patient samples to ensure differentiation is seen in these samples. This system could potentially reduce treatment costs and simplify the treatment strategy for orthopaedic repair and regeneration. PMID:27377664

  7. Comparison of microstructures and phase compositions of artificial and bone-derived hydroxyapatites by transmission electron microscopy and energy dispersive electron spectroscopy.

    PubMed

    Lee, Jong Kook

    2014-11-01

    Biological problems associated with sintered implants fabricated from artificially synthesized hydroxyapatite include selective dissolution at grain boundaries, microstructural disintegration in the body due to particle loosening, and slow crack growth at the implant surface. In addition, mechanical degradation has been shown to be significant, thus limiting their application as load-bearing medical implants. In contrast, bone-derived hydroxyapatite bioceramics have highly dissolution-resistant properties and excellent biocompatibility in the body. They are also easy to synthesize by thermal decomposition of animal bone. In this study, microstructural observations and crystal phase analysis of bone-derived hydroxyapatite were investigated by TEM and EDS using sintered hydroxyapatite samples. In addition, a comparative investigation into elemental distributions and the microstructures of artificial hydroxyapatite, bovine, and tuna bone-derived hydroxyapatites was performed. Bone-derived HA consists mainly of HA and a small amount of MgO. Hot-pressed HA compacts showed homogeneous microstructures and densities of 95-97%, however, grain sizes and microstructures varied with the starting powders. PMID:25958617

  8. Osseointegration aspects of placed implant in bone reconstruction with newly developed block-type interconnected porous calcium hydroxyapatite

    PubMed Central

    DOI, Kazuya; KUBO, Takayasu; MAKIHARA, Yusuke; OUE, Hiroshi; MORITA, Koji; OKI, Yoshifumi; KAJIHARA, Shiho; TSUGA, Kazuhiro

    2016-01-01

    ABSTRACT Artificial bone has been employed to reconstruct bone defects. However, only few reports on implant placement after block bone grafting exist. Objectives The purpose of this study was to evaluate the osseointegration of dental implant in bone reconstructions with interconnected porous calcium hydroxyapatite (IP-CHA). Material and Methods The IP-CHA cylinders (D; 4.3 mm, H; 10.0 mm) were placed into bone sockets in each side of the femurs of four male dogs. The IP-CHA on the right side was a 24-week sample. Twelve weeks after placement, a titanium implant was placed into a socket that was prepared in half of the placed IP-CHA cylinder on the right side. On the left side, another IP-CHA cylinder was placed as a 12-week sample. After another 12 weeks, the samples were harvested, and the bone regeneration and bone-implant contact (BIC) ratios were measured. Results New bone formation area was superior in the 24-week IP-CHA compared with the 12-week IP-CHA. BIC was not significantly different between IP-CHA and the parent sites. Osseointegration was detected around the implant in IP-CHA-reconstructed bone. Conclusion Our preliminary results suggest that IP-CHA may be a suitable bone graft material for reconstructing bones that require implant placement. PMID:27556202

  9. Bone-repair properties of biodegradable hydroxyapatite nano-rod superstructures

    NASA Astrophysics Data System (ADS)

    D'Elía, Noelia L.; Mathieu, Colleen; Hoemann, Caroline D.; Laiuppa, Juan A.; Santillán, Graciela E.; Messina, Paula V.

    2015-11-01

    Nano-hydroxyapatite (nano-HAp) materials show an analogous chemical composition to the biogenic mineral components of calcified tissues and depending on their topography they may mimic the specific arrangement of the crystals in bone. In this work, we have evaluated the potential of four synthesized nano-HAp superstructures for the in vitro conditions of bone-repair. Experiments are underway to investigate the effects of the material microstructure, surface roughness and hydrophilicity on their osseo-integration, osteo-conduction and osteo-induction abilities. Materials were tested in the presence of both, rat primary osteoblasts and rabbit mesenchymal stem cells. The following aspects are discussed: (i) cytotoxicity and material degradation; (ii) rat osteoblast spreading, proliferation and differentiation; and (iii) rabbit mesenchymal stem cell adhesion on nano-HAp and nano-HAp/collagen type I coatings. We effectively prepared a material based on biomimetic HAp nano-rods displaying the appropriate surface topography, hydrophilicity and degradation properties to induce the in vitro desired cellular responses for bone bonding and healing. Cells seeded on the selected material readily attached, proliferated and differentiated, as confirmed by cell viability, mitochondrial metabolic activity, alkaline phosphatase (ALP) activity and cytoskeletal integrity analysis by immunofluorescence localization of alpha-smooth muscle actin (α-SMA) protein. These results highlight the influence of material's surface characteristics to determine their tissue regeneration potential and their future use in engineering osteogenic scaffolds for orthopedic implants.Nano-hydroxyapatite (nano-HAp) materials show an analogous chemical composition to the biogenic mineral components of calcified tissues and depending on their topography they may mimic the specific arrangement of the crystals in bone. In this work, we have evaluated the potential of four synthesized nano-HAp superstructures

  10. Bone Regeneration Using Hydroxyapatite Sponge Scaffolds with In Vivo Deposited Extracellular Matrix.

    PubMed

    Ventura, Reiza Dolendo; Padalhin, Andrew Reyes; Min, Young-Ki; Lee, Byong-Taek

    2015-11-01

    There is currently an increased interest in studying the extracellular matrix (ECM) and its potential applications for tissue engineering and regenerative medicine. The ECM plays an important role by providing adhesive substrates to cells during migration, morphogenesis, differentiation, and homeostasis by signaling biochemical and biomechanical cues to cells. In this study, the ECM was incorporated into hydroxyapatite by implanting sponge replica scaffolds in subcutaneous pockets in rats, and the implants were tested for bone regeneration potential. The resulting scaffolds were characterized using scanning electron microscopy, confocal microscopy, DNA and RNA quantification, tissue staining, energy dispersive X-ray spectroscopy analysis, compressive strength testing, porosity, and pore size distribution analysis using bare scaffolds as a control reference. Biocompatibility was assessed using MC3T3-E1 preosteoblast cells and in vivo studies were carried out by implanting decellularized scaffolds in 11 mm radial defects in New Zealand rabbits for 4 and 8 weeks to determine the effect of the in vivo deposited ECM. Material characterization indicated that a 2-week decellularized scaffold was the best among the samples, with an evenly distributed ECM visible on hematoxylin and eosin-stained tissue sections, a compressive strength of 2.53 ± 0.68 MPa, a porosity of 58.08 ± 3.32% and a pore size distribution range of 10-150 μm. In vivo results showed no severe inflammation, with increased cell infiltration followed by dense matrix deposition after 4 weeks and new bone formation at 8 weeks. The results indicate that incorporation of an in vivo deposited ECM into ceramic scaffolds can potentially improve bone regeneration. PMID:26228909

  11. Treatment of a Simple Bone Cyst Using a Cannulated Hydroxyapatite Pin.

    PubMed

    Shirai, Toshiharu; Tsuchiya, Hiroyuki; Terauchi, Ryu; Tsuchida, Shinji; Mizoshiri, Naoki; Ikoma, Kazuya; Fujiwara, Hiroyoshi; Miwa, Shinji; Kimura, Hiroaki; Takeuchi, Akihiko; Hayashi, Katsuhiro; Yamamoto, Norio; Kubo, Toshikazu

    2015-06-01

    Simple bone cysts (SBCs) are benign bone tumors. However, the treatment of SBCs remains controversial because of their healing rate and the invasiveness of surgery. The purpose of the present study was to evaluate the treatment of SBCs using a cannulated hydroxyapatite (HA) pin.A total of 43 patients (35 males, 8 females; mean age 12.1 years; age range, 5-22 years) with SBCs were treated with continuous decompression by inserting ceramic HA pins between 1989 and 2014. The SBCs were located in the calcaneus in 23, the humerus in 15, the femur in 3, and the pelvis in 2 cases. In all patients, minimal fenestration of the cyst wall and curettage and multiple drilling in the cyst wall were performed, followed by insertion of the HA pin. The mean follow-up period was 26.6 months. Operating time, healing period, risk factors for recurrence, and the cure rate were evaluated.Healing was achieved without intervention in 38 patients after a mean of 6.4 months. Two patients had persistent small residual cysts, which had no changes after 1 year at the latest follow-up. There were 5 patients with recurrences (humerus 4, femur 1), who were cured by curettage and artificial bone grafting. The final healing rate by cannulation only using an HA pin was 88.2%. On Fisher exact test, age, site of SBCs, and distance from the physis were found to be significantly associated with SBC recurrence (P < 0.05).In the present study, cannulation using an HA pin for SBCs was found to be a useful technique, particularly for calcaneal cysts, because it is a minimally invasive procedure with a high cure rate. In patients <10 years, involvement of the humerus and contact with the growth plate were significant risk factors for SBC recurrence. PMID:26107670

  12. Treatment of a Simple Bone Cyst Using a Cannulated Hydroxyapatite Pin

    PubMed Central

    Shirai, Toshiharu; Tsuchiya, Hiroyuki; Terauchi, Ryu; Tsuchida, Shinji; Mizoshiri, Naoki; Ikoma, Kazuya; Fujiwara, Hiroyoshi; Miwa, Shinji; Kimura, Hiroaki; Takeuchi, Akihiko; Hayashi, Katsuhiro; Yamamoto, Norio; Kubo, Toshikazu

    2015-01-01

    Abstract Simple bone cysts (SBCs) are benign bone tumors. However, the treatment of SBCs remains controversial because of their healing rate and the invasiveness of surgery. The purpose of the present study was to evaluate the treatment of SBCs using a cannulated hydroxyapatite (HA) pin. A total of 43 patients (35 males, 8 females; mean age 12.1 years; age range, 5–22 years) with SBCs were treated with continuous decompression by inserting ceramic HA pins between 1989 and 2014. The SBCs were located in the calcaneus in 23, the humerus in 15, the femur in 3, and the pelvis in 2 cases. In all patients, minimal fenestration of the cyst wall and curettage and multiple drilling in the cyst wall were performed, followed by insertion of the HA pin. The mean follow-up period was 26.6 months. Operating time, healing period, risk factors for recurrence, and the cure rate were evaluated. Healing was achieved without intervention in 38 patients after a mean of 6.4 months. Two patients had persistent small residual cysts, which had no changes after 1 year at the latest follow-up. There were 5 patients with recurrences (humerus 4, femur 1), who were cured by curettage and artificial bone grafting. The final healing rate by cannulation only using an HA pin was 88.2%. On Fisher exact test, age, site of SBCs, and distance from the physis were found to be significantly associated with SBC recurrence (P < 0.05). In the present study, cannulation using an HA pin for SBCs was found to be a useful technique, particularly for calcaneal cysts, because it is a minimally invasive procedure with a high cure rate. In patients <10 years, involvement of the humerus and contact with the growth plate were significant risk factors for SBC recurrence. PMID:26107670

  13. Chemical and physicochemical characterization of porous hydroxyapatite ceramics made of natural bone.

    PubMed

    Joschek, S; Nies, B; Krotz, R; Göferich, A

    2000-08-01

    The properties of a porous hydroxyapatite ceramic produced by sintering of bovine bone were investigated by using a number of physicochemical methods such as scanning electron microscopy (SEM), SEM in combination with energy dispersive X-ray spectroscopy (SEM-EDX), mercury intrusion porosimetry, krypton-adsorption, contact angle measurements, wide angle X-ray diffraction. Fourier transform infrared spectroscopy, thermal analysis, inductively coupled plasma optical atom emissions spectroscopy and flame atomic absorption spectroscopy. The results indicate that there are considerable differences between the ceramic and native bone. However, the most important properties with respect to the use of such ceramics as a biomaterial for filling bone defects namely the high porosity (> or = 57 +/- 2%) and the interconnecting pore system are maintained. While macropores with an average diameter of approx. 300 microm contribute 97% to porosity, micropores with an average diameter of 1.3 microm account for only 3% of the total porosity. The surface area was found to be approx. 0.1 m2/g. The contact angles of water (44.6 +/- 15.4 degrees, n = 5) and tetrahydrofurane (10 degrees) allow the processing of the ceramic to a drug carrier by incubation with aqueous or organic drug solutions. The ceramic is highly crystalline with crystal sizes of 1-7 microm and contains crystal bridges. The investigation of its chemical composition revealed small amounts of other inorganic compounds such as Ca4O(PO4)2, NaCaPO4, Ca3(PO4)2, CaO, and MgO. Besides trace amounts of aluminum, iron, magnesium, potassium, silica, sodium, vanadium and zinc it contains probably carbonated apatite. PMID:10905406

  14. Bone integration capability of nanopolymorphic crystalline hydroxyapatite coated on titanium implants

    PubMed Central

    Yamada, Masahiro; Ueno, Takeshi; Tsukimura, Naoki; Ikeda, Takayuki; Nakagawa, Kaori; Hori, Norio; Suzuki, Takeo; Ogawa, Takahiro

    2012-01-01

    The mechanism by which hydroxyapatite (HA)-coated titanium promotes bone–implant integration is largely unknown. Furthermore, refining the fabrication of nano-structured HA to the level applicable to the mass production process for titanium implants is challenging. This study reports successful creation of nanopolymorphic crystalline HA on microroughened titanium surfaces using a combination of flame spray and low-temperature calcination and tests its biological capability to enhance bone–implant integration. Sandblasted microroughened titanium implants and sandblasted + HA-coated titanium implants were subjected to biomechanical and histomorphometric analyses in a rat model. The HA was 55% crystallized and consisted of nanoscale needle-like architectures developed in various diameters, lengths, and orientations, which resulted in a 70% increase in surface area compared to noncoated microroughened surfaces. The HA was free from impurity contaminants, with a calcium/phosphorus ratio of 1.66 being equivalent to that of stoichiometric HA. As compared to microroughened implants, HA-coated implants increased the strength of bone–implant integration consistently at both early and late stages of healing. HA-coated implants showed an increased percentage of bone–implant contact and bone volume within 50 μm proximity of the implant surface, as well as a remarkably reduced percentage of soft tissue intervention between bone and the implant surface. In contrast, bone volume outside the 50 μm border was lower around HA-coated implants. Thus, this study demonstrated that the addition of pure nanopolymorphic crystalline HA to microroughened titanium not only accelerates but also enhances the level of bone–implant integration and identified the specific tissue morphogenesis parameters modulated by HA coating. In particular, the nanocrystalline HA was proven to be drastic in increasing osteoconductivity and inhibiting soft tissue infiltration, but the effect was

  15. Physicochemical study of plasma-sprayed hydroxyapatite-coated implants in humans.

    PubMed

    MacDonald, D E; Betts, F; Stranick, M; Doty, S; Boskey, A L

    2001-03-15

    This study represents the first report of the physical and chemical changes occurring in coatings of failed hydroxyapatite (HA)-coated titanium implants obtained from a comprehensive, multicenter human dental implant study. A total of 53 retrieved samples were obtained and compared with unimplanted controls with the same manufacturer and similar manufacture dates. Forty-five retrieved implants were examined for surface characteristics and bulk composition. Implants were staged based on implantation history: stage 1 (implants retrieved between surgical placement and surgical uncovering), stage 2 (implants retrieved at surgical uncovering and evaluation), stage 3 (implants retrieved between surgical uncovering evaluation and occlusal loading), and stage 4 (implants retrieved after occlusal loading). Scanning electron microscopy showed progressive coating thinning with implantation time. At later stages, bare Ti metal was detected by energy-dispersive X-ray analysis and electron spectroscopy for chemical analysis. Increases in Ti and Al (2-7.5 atm % each) were detected at the apical ends of all stage 4 samples. In unimplanted coatings, X-ray diffraction analysis demonstrated the presence of amorphous calcium phosphate, beta-tricalcium phosphate, tetracalcium phosphate, and calcium oxide in addition to large hydroxyapatite crystals (c axis size, D002 = 429 +/- 13 A; a axis size, D300 = 402 +/- 11 A, a/c aspect ratio 0.92). The nonapatitic phases disappeared with increased implantation time, although there was a persistence of amorphous calcium phosphate. Bulk coating chemical analysis showed that Ca/P ratios for implant controls (1.81 +/- 0.01) were greater than stoichiometric HA (1.67) and decreased for implant stages 3 and 4 (1.69 +/- 0.09 and 1.67 +/- 0.09, respectively), explained by the dissolution of the non apatitic phases. Crystal sizes also changed with implantation times, being smaller than the control at all but stage 4. Fourier transform infrared analyses

  16. Effect of incorporation of nanoscale bioactive glass and hydroxyapatite in PCL/chitosan nanofibers for bone and periodontal tissue engineering.

    PubMed

    Shalumon, K T; Sowmya, S; Sathish, D; Chennazhi, K P; Nair, Shantikumar V; Jayakumar, R

    2013-03-01

    A biomimetic scaffold which can very closely mimic the extracellular matrix of the bone was fabricated by incorporating nano-bioceramic particles such as nano bioglass (nBG) and nano hydroxyapatite (nHAp) within electrospun nanofibrous scaffold. A comparative study between nHAp incorporated poly(caprolactone) (PCL)-chitosan (CS) and nBG incorporated PCL-CS nanofibrous scaffolds was carried out and their feasibility in tissue engineering was investigated. All the samples were optimized to obtain fibers of similar diameter from 100-200 nm for the ease of comparison between the samples. Protein adsorption studies showed that PCL-CS incorporated with 3 wt% nHAp and 3 wt% nBG adsorbed more proteins on their surface than other samples. Cell attachment and proliferation studies using human periodontal ligament fibroblast cells (hPLFs) and osteoblast like cells (MG-63 cell lines) showed that nBG incorporated samples are slightly superior to nHAp incorporated counterparts. Cell viability test using alamar blue assay and live/dead staining confirms that the scaffolds are cytocompatible. ALP activity confirmed the osteoblastic behavior of hPDLFs. Also the presence of nHAp and nBG enhanced the ALP activity of hPDLF on the PCH3 and PCB3 scaffolds. These studies indicate that nBG incorporated electrospun scaffolds are comparatively better candidates for orthopedic and periodontal tissue engineering applications. PMID:23620999

  17. Thermoplastic polyurethane/hydroxyapatite electrospun scaffolds for bone tissue engineering: effects of polymer properties and particle size.

    PubMed

    Mi, Hao-Yang; Palumbo, SunMi; Jing, Xin; Turng, Lih-Sheng; Li, Wan-Ju; Peng, Xiang-Fang

    2014-10-01

    Thermoplastic polyurethane (TPU)/hydroxyapatite (HA) scaffolds were fabricated via electrospinning. The effects of TPU properties and HA particle size on scaffold physical properties and osteoblast-like cell performance were investigated. It was found that the addition of micro-HA (mHA), which was inlayed in the fiber, decreased the electrospun fiber diameter. On the contrary, nano-HA (nHA), which was either embedded or existed inside of the fiber, increased the fiber diameter for both soft and hard TPUs. The soft TPU had a much lower Young's modulus and higher strain-at-break than the hard TPU. The addition of both mHA and nHA decreased the tensile properties; this decrease was more significant with mHA. The cells on the hard scaffolds actively proliferated and migrated compared to those on the soft scaffolds. On the other hand, cells on the soft scaffolds more effectively induced osteogenesis of human mesenchymal stem cells (hMSCs) than those on the hard scaffolds. In addition, our data suggest that the soft scaffolds with supplementation of nHA further enhanced osteogenesis of hMSCs compared to those without nHA. The soft TPU scaffolds containing nano-HA have the potential to be used in bone tissue engineering applications. PMID:24574168

  18. Electrospun composites of PHBV, silk fibroin and nano-hydroxyapatite for bone tissue engineering.

    PubMed

    Paşcu, Elena I; Stokes, Joseph; McGuinness, Garrett B

    2013-12-01

    Electrospinning of fibrous scaffolds containing nano-hydroxyapatite (nHAp) embedded in a matrix of functional biomacromolecules offers an attractive route to mimicking the natural bone tissue architecture. Functional fibrous substrates will support cell attachment, proliferation and differentiation, while the role of HAp is to induce cells to secrete extracellular matrix (ECM) for mineralization to form bone. Electrospinning of biomaterials composed of polyhydroxybutyrate-co-(3-hydroxyvalerate) with 2% valerate fraction (PHBV), nano-hydroxyapatite (nHAp), and Bombyx mori silk fibroin essence (SF), Mw=90KDa, has been achieved for nHAp and SF solution concentrations of 2 (w/vol) % each and 5 (w/vol) % each. The structure and properties of the nanocomposite fibrous membranes were investigated by means of Scanning Electron Microscopy in combination with Energy Dispersive X-Ray Analysis (SEM/EDX), Fourier Transformed Infrared Spectroscopy (FT-IR), uniaxial tensile and compressive mechanical testing, degradation tests and in vitro bioactivity tests. SEM images showed smooth, uniform and continuous fibre deposition with no bead formation, and fibre diameters of between 10 and 15 μm. EDX and FT-IR confirmed the presence of nHAp and SF. After one month in deionised water, tests showed less than 2% weight loss with the samples retaining their fibrous morphology, confirming that this material biodegrades slowly. After 28 days of immersion in Simulated Body Fluid (SBF) an apatite layer was visible on the surface of the fibres, proving their bioactivity. Preliminary in vitro biological assessment showed that after 1 and 3 days in culture, cells were attached to the fibres, retaining their morphology while presenting a flattened appearance and elongated shape on the surface of fibres. Young's modulus was found to increase from 0.7 kPa (±0.33 kPa) for electrospun samples of PHBV only to 1.4 kPa (±0.54 kPa) for samples with 2 (w/vol) % each of nHAp and SF. Samples prepared with

  19. Graphene and hydroxyapatite self-assemble into homogeneous, free standing nanocomposite hydrogels for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Xie, Xingyi; Hu, Kaiwen; Fang, Dongdong; Shang, Lihong; Tran, Simon D.; Cerruti, Marta

    2015-04-01

    Graphene-nanoparticle (NP) composites have shown potential in applications ranging from batteries to, more recently, tissue engineering. Graphene and NPs should be integrated into uniform free-standing structures for best results. However, to date, this has been achieved only in few examples; in most cases, graphene/NP powders lacking three-dimensional (3D) structure were produced. Here we report a facile and universal method that can be used to synthesize such structures based on colloidal chemistry. We start from aqueous suspensions of both graphene oxide nanosheets and citrate-stabilized hydroxyapatite (HA) NPs. Hydrothermal treatment of the mixtures of both suspensions reduces graphene oxide to graphene, and entraps colloidal HA NPs into the 3D graphene network thanks to a self-assembled graphite-like shell formed around it. Dialysis through this shell causes uniform NP deposition onto the graphene walls. The resulting graphene-HA gels are highly porous, strong, electrically conductive and biocompatible, making them promising scaffolds for bone tissue engineering. This method can be applied to produce a variety of free-standing 3D graphene-based nanocomposites with unprecedented homogeneity.Graphene-nanoparticle (NP) composites have shown potential in applications ranging from batteries to, more recently, tissue engineering. Graphene and NPs should be integrated into uniform free-standing structures for best results. However, to date, this has been achieved only in few examples; in most cases, graphene/NP powders lacking three-dimensional (3D) structure were produced. Here we report a facile and universal method that can be used to synthesize such structures based on colloidal chemistry. We start from aqueous suspensions of both graphene oxide nanosheets and citrate-stabilized hydroxyapatite (HA) NPs. Hydrothermal treatment of the mixtures of both suspensions reduces graphene oxide to graphene, and entraps colloidal HA NPs into the 3D graphene network thanks to

  20. Recombinant human bone morphogenetic protein induces bone formation.

    PubMed Central

    Wang, E A; Rosen, V; D'Alessandro, J S; Bauduy, M; Cordes, P; Harada, T; Israel, D I; Hewick, R M; Kerns, K M; LaPan, P

    1990-01-01

    We have purified and characterized active recombinant human bone morphogenetic protein (BMP) 2A. Implantation of the recombinant protein in rats showed that a single BMP can induce bone formation in vivo. A dose-response and time-course study using the rat ectopic bone formation assay revealed that implantation of 0.5-115 micrograms of partially purified recombinant human BMP-2A resulted in cartilage by day 7 and bone formation by day 14. The time at which bone formation occurred was dependent on the amount of BMP-2A implanted; at high doses bone formation could be observed at 5 days. The cartilage- and bone-inductive activity of the recombinant BMP-2A is histologically indistinguishable from that of bone extracts. Thus, recombinant BMP-2A has therapeutic potential to promote de novo bone formation in humans. Images PMID:2315314

  1. Angiogenesis and bone regeneration of porous nano-hydroxyapatite/coralline blocks coated with rhVEGF165 in critical-size alveolar bone defects in vivo

    PubMed Central

    Du, Bing; Liu, Weizhen; Deng, Yue; Li, Shaobing; Liu, Xiangning; Gao, Yan; Zhou, Lei

    2015-01-01

    To improve the regenerative performance of nano-hydroxyapatite/coralline (nHA/coral) block grafting in a canine mandibular critical-size defect model, nHA/coral blocks were coated with recombinant human vascular endothelial growth factor165 (rhVEGF) via physical adsorption (3 μg rhVEGF165 per nHA/coral block). After the nHA/coral blocks and VEGF/nHA/coral blocks were randomly implanted into the mandibular box-shaped defects in a split-mouth design, the healing process was evaluated by histological observation and histomorphometric and immunohistological analyses. The histological evaluations revealed the ingrowth of newly formed blood vessels and bone at the periphery and cores of the blocks in both groups at both 3 and 8 weeks postsurgery, respectively. In the histomorphometric analysis, the VEGF/nHA/coral group exhibited a larger quantity of new bone formation at 3 and 8 weeks postsurgery. The percentages of newly formed bone within the entire blocks in the VEGF/nHA/coral group were 27.3%±8.1% and 39.3%±12.8% at 3 weeks and 8 weeks, respectively, and these values were slightly greater than those of the nHA/coral group (21.7%±3.0% and 32.6%±10.3%, respectively), but the differences were not significant (P>0.05). The immunohistological evaluations revealed that the neovascular density in the VEGF/nHA/coral group (146±32.9 vessel/mm2) was much greater than that in the nHA/coral group (105±51.8 vessel/mm2) at the 3-week time point (P<0.05), but no significant difference was observed at the 8-week time point (341±86.1 and 269±50.7 vessel/mm2, respectively, P>0.05). The present study indicated that nHA/coral blocks might be optimal scaffolds for block grafting in critical-size mandibular defects and that additional VEGF coating via physical adsorption can promote angiogenesis in the early stage of bone healing, which suggests that prevascularized nHA/coral blocks have significant potential as a bioactive material for bone regeneration in large-scale alveolar

  2. Development of biomimetic nanocomposites as bone extracellular matrix for human osteoblastic cells.

    PubMed

    Bhowmick, Arundhati; Mitra, Tapas; Gnanamani, Arumugam; Das, Manas; Kundu, Patit Paban

    2016-05-01

    Here, we have developed biomimetic nanocomposites containing chitosan, poly(vinyl alcohol) and nano-hydroxyapatite-zinc oxide as bone extracellular matrix for human osteoblastic cells and characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction. Scanning electron microscopy images revealed interconnected macroporous structures. Moreover, in this study, the problem related to fabricating a porous composite with good mechanical strength has been resolved by incorporating 5wt% of nano-hydroxyapatite-zinc oxide into chitosan-poly(vinyl alcohol) matrix; the present composite showed high tensile strength (20.25MPa) while maintaining appreciable porosity (65.25%). These values are similar to human cancellous bone. These nanocomposites also showed superior water uptake, antimicrobial and biodegradable properties than the previously reported results. Compatibility with human blood and pH was observed, indicating nontoxicity of these materials to the human body. Moreover, proliferation of osteoblastic MG-63 cells onto the nanocomposites was also observed without having any negative effect. PMID:26876999

  3. Titania-hydroxyapatite nanocomposite coatings support human mesenchymal stem cells osteogenic differentiation.

    PubMed

    Dimitrievska, Sashka; Bureau, Martin N; Antoniou, John; Mwale, Fackson; Petit, Alain; Lima, Rogerio S; Marple, Basil R

    2011-09-15

    In addition to mechanical and chemical stability, the third design goal of the ideal bone-implant coating is the ability to support osteogenic differentiation of mesenchymal stem cells (MSCs). Plasma-sprayed TiO(2)-based bone-implant coatings exhibit excellent long-term mechanical properties, but their applications in bone implants are limited by their bioinertness. We have successfully produced a TiO(2) nanostructured (grain size <50 nm) based coating charged with 10% wt hydroxyapatite (TiO(2)-HA) sprayed by high-velocity oxy-fuel. On Ti64 substrates, the novel TiO(2)-HA coating bond 153× stronger and has a cohesive strength 4× higher than HA coatings. The HA micro- and nano-sized particles covering the TiO(2)-HA coating surface are chemically bound to the TiO(2) coating matrix, producing chemically stable coatings under high mechanical solicitations. In this study, we elucidated the TiO(2)-HA nanocomposite coating surface chemistry, and in vitro osteoinductive potential by culturing human MSCs (hMSCs) in basal and in osteogenic medium (hMSC-ob). We assessed the following hMSCs and hMSC-ob parameters over a 3-week period: (i) proliferation; (ii) cytoskeleton organization and cell-substrate adhesion; (iii) coating-cellular interaction morphology and growth; and (iv) cellular mineralization. The TiO(2) -HA nanocomposite coatings demonstrated 3× higher hydrophilicity than HA coatings, a TiO(2)-nanostructured surface in addition to the chemically bound HA micron- and nano-sized rod to the surface. hMSCs and hMSC-ob demonstrated increased proliferation and osteoblastic differentiation on the nanostructured TiO(2)-HA coatings, suggesting the TiO(2)-HA coatings nanostructure surface properties induce osteogenic differentiation of hMSC and support hMSC-ob osteogenic potential better than our current golden standard HA coating. PMID:21702080

  4. Hydroxyapatite surface roughness: complex modulation of the osteoclastogenesis of human precursor cells.

    PubMed

    Costa-Rodrigues, João; Fernandes, Anabela; Lopes, Maria A; Fernandes, Maria H

    2012-03-01

    It is recognized that the surface roughness affects osteoblastic differentiation, but little information is available regarding its effect on osteoclastogenesis. With this work, the osteoclastogenic behaviour of human peripheral blood mononuclear cells (PBMCs), cultured isolated (1.5×10(6)cellscm(-2)) or co-cultured with human bone marrow cells (hBMCs; 10(3)cellscm(-2)), was assessed on surface-abraded hydroxyapatite disks with three different surface roughnesses (R(a) 0.0437-0.582 μm). Monocultures and co-cultures were performed for 21 days in the absence or presence of recombinant M-CSF and RANKL. Results showed that PBMCs supplemented with M-CSF and RANKL or co-cultured with hBMCs displayed typical osteoclastic features, i.e. multinucleated cells with actin rings, vitronectin and calcitonin receptors, gene expression of TRAP, cathepsin K, carbonic anhydrase 2, c-myc and c-src, TRAP activity and resorbing activity. The osteoclastogenic response increased with surface roughness in PBMCs cultured with M-CSF and RANKL but decreased in PBMCs co-cultured with hBMCs. However, co-cultures supplemented with the osteoclastogenic inducers displayed high and similar levels of osteoclast differentiation in the three tested surfaces. In conclusion, modulation of osteoclast differentiation by surface roughness seemed to be dependent on the mechanisms subjacent to the osteoclastogenic stimulus, i.e. the presence of soluble factors or direct cell-to-cell contacts between osteoblastic and osteoclastic cells. PMID:22178652

  5. Mechanical and biological properties of the micro-/nano-grain functionally graded hydroxyapatite bioceramics for bone tissue engineering.

    PubMed

    Zhou, Changchun; Deng, Congying; Chen, Xuening; Zhao, Xiufen; Chen, Ying; Fan, Yujiang; Zhang, Xingdong

    2015-08-01

    Functionally graded materials (FGM) open the promising approach for bone tissue repair. In this study, a novel functionally graded hydroxyapatite (HA) bioceramic with micrograin and nanograin structure was fabricated. Its mechanical properties were tailored by composition of micrograin and nanograin. The dynamic mechanical analysis (DMA) indicated that the graded HA ceramics had similar mechanical property compared to natural bones. Their cytocompatibility was evaluated via fluorescent microscopy and MTT colorimetric assay. The viability and proliferation of rabbit bone marrow mesenchymal stem cells (BMSCs) on ceramics indicated that this functionally graded HA ceramic had better cytocompatibility than conventional HA ceramic. This study demonstrated that functionally graded HA ceramics create suitable structures to satisfy both the mechanical and biological requirements of bone tissues. PMID:25910818

  6. Effect of cyclic loading on the nanoscale deformation of hydroxyapatite and collagen fibrils in bovine bone.

    PubMed

    Singhal, Anjali; Stock, Stuart R; Almer, Jonathan D; Dunand, David C

    2014-06-01

    Cyclic compressive loading tests were carried out on bovine femoral bones at body temperature (37 °C), with varying mean stresses (-55 to -80 MPa) and loading frequencies (0.5-5 Hz). At various times, the cyclic loading was interrupted to carry out high-energy X-ray scattering measurements of the internal strains developing in the hydroxyapatite (HAP) platelets and the collagen fibrils. The residual strains upon unloading were always tensile in the HAP and compressive in the fibrils, and each increases in magnitude with loading cycles, which can be explained from damage at the HAP–collagen interface and accumulation of plastic deformation within the collagen phase. The samples tested at a higher mean stress and stress amplitude, and at lower loading frequencies exhibit greater plastic deformation and damage accumulation, which is attributed to greater contribution of creep. Synchrotron microcomputed tomography of some of the specimens showed that cracks are produced during cyclic loading and that they mostly occur concentric with Haversian canals. PMID:23958833

  7. Bioactive nanocomposite PLDL/nano-hydroxyapatite electrospun membranes for bone tissue engineering.

    PubMed

    Rajzer, Izabella; Menaszek, Elżbieta; Kwiatkowski, Ryszard; Chrzanowski, Wojciech

    2014-05-01

    New nanocomposite membranes with high bioactivity were fabricated using the electrospinning. These nanocomposites combine a degradable polymer poly(L/DL)-lactide and bone cell signaling carbonate nano-hydroxyapatite (n-HAp). Chemical and physical characterization of the membranes using scanning electron microscopy, Fourier transform infrared spectroscopy and the wide angle X-ray diffraction evidenced that nanoparticles were successfully incorporated into the fibers and membrane structure. The incorporation of the n-HAp into the structure increased significantly the mineralization of the membrane in vitro. It has been demonstrated that after a 3-day incubation of composite membrane in the Simulated Body Fluid a continuous compact apatite layer was formed. In vitro experiments demonstrated that the incorporation of n-HAp significantly improved cell attachment, upregulated cells proliferation and stimulated cell differentiation quantified using Alkaline Phosphatase and OsteoImage tests. In conclusion, the results demonstrated that the addition of n-HAp provided chemical cues that were a key factor that regulated osteoblastic differentiation. PMID:24458535

  8. A novel two-step sintering for nano-hydroxyapatite scaffolds for bone tissue engineering

    PubMed Central

    Feng, Pei; Niu, Man; Gao, Chengde; Peng, Shuping; Shuai, Cijun

    2014-01-01

    In this study, nano-hydroxyapatite scaffolds with high mechanical strength and an interconnected porous structure were prepared using NTSS for the first time. The first step was performed using a laser characterized by the rapid heating to skip the surface diffusion and to obtain the driving force for grain boundary diffusion. Additionally, the interconnected porous structure was achieved by SLS. The second step consisted of isothermal heating in a furnace at a lower temperature (T2) than that of the laser beam to further increase the density and to suppress grain growth by exploiting the difference in kinetics between grain-boundary diffusion and grain-boundary migration. The results indicated that the mechanical properties first increased and then decreased as T2 was increased from 1050 to 1250°C. The optimal fracture toughness, compressive strength and stiffness were 1.69 MPam1/2, 18.68 MPa and 245.79 MPa, respectively. At the optimal point, the T2 was 1100°C, the grain size was 60 nm and the relative density was 97.6%. The decrease in mechanical properties was due to the growth of grains and the decomposition of HAP. The cytocompatibility test results indicated that cells adhered and spread well on the scaffolds. A bone-like apatite layer formed, indicating good bioactivity. PMID:24998362

  9. 3D printing of porous hydroxyapatite scaffolds intended for use in bone tissue engineering applications.

    PubMed

    Cox, Sophie C; Thornby, John A; Gibbons, Gregory J; Williams, Mark A; Mallick, Kajal K

    2015-02-01

    A systematic characterisation of bone tissue scaffolds fabricated via 3D printing from hydroxyapatite (HA) and poly(vinyl)alcohol (PVOH) composite powders is presented. Flowability of HA:PVOH precursor materials was observed to affect mechanical stability, microstructure and porosity of 3D printed scaffolds. Anisotropic behaviour of constructs and part failure at the boundaries of interlayer bonds was highlighted by compressive strength testing. A trade-off between the ability to facilitate removal of PVOH thermal degradation products during sintering and the compressive strength of green parts was revealed. The ultimate compressive strength of 55% porous green scaffolds printed along the Y-axis and dried in a vacuum oven for 6h was 0.88 ± 0.02 MPa. Critically, the pores of 3D printed constructs could be user designed, ensuring bulk interconnectivity, and the imperfect packing of powder particles created an inherent surface roughness and non-designed porosity within the scaffold. These features are considered promising since they are known to facilitate osteoconduction and osteointegration in-vivo. Characterisation techniques utilised in this study include two funnel flow tests, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), compressive strength testing and computed tomography (CT). PMID:25492194

  10. Poly (D,L-lactide)/nano-hydroxyapatite composite scaffolds for bone tissue engineering and biocompatibility evaluation.

    PubMed

    Ren, Jie; Zhao, Peng; Ren, Tianbin; Gu, Shuying; Pan, Kefeng

    2008-03-01

    Biodegradable polymer/bioceramic composite scaffolds can overcome the limitations of conventional ceramic bone substitutes such as brittleness and difficulty in shaping. However, conventional methods for fabricating polymer/bioceramic composite scaffolds often use organic solvents (e.g., the solvent casting and particulate leaching (SC/PL) method), which might be harmful to cells or tissues. In this study, Poly (D,L-lactide)/nano-hydroxyapatite (PDLLA/NHA) composites were prepared by in-situ polymerization, and highly porous scaffolds were fabricated using a novel method, supercritical CO2/salt-leaching method (SC CO2/SL). The materials and scaffolds were investigated by scanning electronic microscopy (SEM), transmission electronic microscopy (TEM) and gel permeation chromatography (GPC). GPC showed that the molecular weight of composites decreased with increase of NHA content. However, the water absorption and compressive strength increased dramatically. The SEM micrographs showed that the scaffolds with pore size about 250 microm were obtained by controlling parameters of SC CO2/SL. The biocompatibility of PDLLA/NHA porous scaffolds were evaluated in vitro and in vivo. The evaluation on the cytotoxicity were carried out by cell relative growth rate (RGR) method and cell direct contact method. The cytotoxicity of these scaffolds was in grade I according to ISO 10993-1. There was no toxicosis and death cases observed in acute systemic toxicity test. And histological observation of the tissue response (1 and 9 weeks after the implantation) showed that there are still some slight inflammation responses. PMID:17701303

  11. Study of bone-like hydroxyapatite/polyamino acid composite materials for their biological properties and effects on the reconstruction of long bone defects

    PubMed Central

    Yan, Ling; Jiang, Dian-ming

    2015-01-01

    Purpose The purpose of this study was to investigate the effect of bone-like hydroxyapatite/polyamino acid (BHA/PAA) in the osteogenesis and reconstruction of long segmental bone defects. Methods In vitro, MG63 cells were cultured with BHA/PAA. The osteoinductive activity of the BHA/PAA material was evaluated using inverted microscopy, scanning electron microscopy, MTT proliferation assay, and the determination of alkaline phosphatase activity and Ca2+ content. In vivo, the radial bone defect was made in 20 New Zealand White rabbits, and then these animal were randomly divided into two groups (n=10), the experimental group (with BHA/PAA) and the control group (without BHA/PAA). Postoperatively, the osteogenesis effect of BHA/PAA was evaluated through X-ray, hematoxylin–eosin staining, observation of the gross bone specimen, immunohistochemistry, and fluorescent confocal scanning microscopy. Results In vitro, BHA/PAA promoted the adhesion, growth, and calcium nodule formation of MG63 cells, and it had good osteogenesis activity. In vivo, with BHA/PAA material degradation and absorption, the new bone gradually formed, and the bone defect gradually recovered in the experimental group. In the control group, a limited bone formation was found at the bone broken ends, and the bone defect was obviously visible. Conclusion In vitro and in vivo, we confirmed that BHA/PAA was effective in inducing osteogenesis and reconstructing a long segmental bone defect. PMID:26719675

  12. Mode of heparin attachment to nanocrystalline hydroxyapatite affects its interaction with bone morphogenetic protein-2.

    PubMed

    Goonasekera, Chandhi S; Jack, Kevin S; Bhakta, Gajadhar; Rai, Bina; Luong-Van, Emma; Nurcombe, Victor; Cool, Simon M; Cooper-White, Justin J; Grøndahl, Lisbeth

    2015-01-01

    Heparin has a high affinity for bone morphogenetic protein-2 (BMP-2), which is a key growth factor in bone regeneration. The aim of this study was to investigate how the rate of release of BMP-2 was affected when adsorbed to nanosized hydroxyapatite (HAP) particles functionalized with heparin by different methods. Heparin was attached to the surface of HAP, either via adsorption or covalent coupling, via a 3-aminopropyltriethoxysilane (APTES) layer. The chemical composition of the particles was evaluated using X-ray photoelectron spectroscopy and elemental microanalysis, revealing that the heparin grafting densities achieved were dependent on the curing temperature used in the fabrication of APTES-modified HAP. Comparable amounts of heparin were attached via both covalent coupling and adsorption to the APTES-modified particles, but characterization of the particle surfaces by zeta potential and Brunauer-Emmett-Teller measurements indicated that the conformation of the heparin on the surface was dependent on the method of attachment, which in turn affected the stability of heparin on the surface. The release of BMP-2 from the particles after 7 days in phosphate-buffered saline found that 31% of the loaded BMP-2 was released from the APTES-modified particles with heparin covalently attached, compared to 16% from the APTES-modified particles with the heparin adsorbed. Moreover, when heparin was adsorbed onto pure HAP, it was found that the BMP-2 released after 7 days was 5% (similar to that from unmodified HAP). This illustrates that by altering the mode of attachment of heparin to HAP the release profile and total release of BMP-2 can be manipulated. Importantly, the BMP-2 released from all the heparin particle types was found by the SMAD 1/5/8 phosphorylation assay to be biologically active. PMID:26474791

  13. Porous biomorphic silicon carbide ceramics coated with hydroxyapatite as prospective materials for bone implants.

    PubMed

    Gryshkov, Oleksandr; Klyui, Nickolai I; Temchenko, Volodymyr P; Kyselov, Vitalii S; Chatterjee, Anamika; Belyaev, Alexander E; Lauterboeck, Lothar; Iarmolenko, Dmytro; Glasmacher, Birgit

    2016-11-01

    Porous and cytocompatible silicon carbide (SiC) ceramics derived from wood precursors and coated with bioactive hydroxyapatite (HA) and HA-zirconium dioxide (HA/ZrO2) composite are materials with promising application in engineering of bone implants due to their excellent mechanical and structural properties. Biomorphic SiC ceramics have been synthesized from wood (Hornbeam, Sapele, Tilia and Pear) using a forced impregnation method. The SiC ceramics have been coated with bioactive HA and HA/ZrO2 using effective gas detonation deposition approach (GDD). The surface morphology and cytotoxicity of SiC ceramics as well as phase composition and crystallinity of deposited coatings were analyzed. It has been shown that the porosity and pore size of SiC ceramics depend on initial wood source. The XRD and FTIR studies revealed the preservation of crystal structure and phase composition of in the HA coating, while addition of ZrO2 to the initial HA powder resulted in significant decomposition of the final HA/ZrO2 coating and formation of other calcium phosphate phases. In turn, NIH 3T3 cells cultured in medium exposed to coated and uncoated SiC ceramics showed high re-cultivation efficiency as well as metabolic activity. The recultivation efficiency of cells was the highest for HA-coated ceramics, whereas HA/ZrO2 coating improved the recultivation efficiency of cells as compared to uncoated SiC ceramics. The GDD method allowed generating homogeneous HA coatings with no change in calcium to phosphorus ratio. In summary, porous and cytocompatible bio-SiC ceramics with bioactive coatings show a great promise in construction of light, robust, inexpensive and patient-specific bone implants for clinical application. PMID:27524006

  14. Reverse micelle mediated synthesis, processing, mechanical and biological characterization of hydroxyapatite nanopowders for bone graft application

    NASA Astrophysics Data System (ADS)

    Banerjee, Ashis

    Hydroxyapatite (HA) is the most widely used bioceramic material in bone graft applications because of its compositional similarity with natural bone. However, synthetic HA does not show similar mechanical and biological properties to the inorganic component of bone. Properties of ceramic material depend on starting materials, processing techniques, densification and microstructure of the final product. The objective of this research was to process HA whisker reinforced HA composite using HA nanopowders and whiskers. HA nanopowders with different length scale and morphology were synthesized by reverse micelle system using NP5 and NP12 as surfactants and cyclohexane as organic solvent. The lowest average aspect ratio was 1.357+/-0.39 with average particle size of 66 nm and the highest average aspect ratio was 7.277+/-3.227 with average length of 150 nm and width of 20 nm, were synthesized. Micron sized HA whiskers with aspect ratio between 20 and 50, average particle length of 15 mum and width of 400 nm was synthesized using urea as a precipitating agent. Desired microstructure was obtained after sintering with spherical HA nanopowder and whiskers along with dopants. Addition of whiskers decreased density of the sintered compacts. However, at 10 wt% whisker content sample showed microhardness and fracture toughness of 3.6 GPa and 1.5 MPa.m1/2, respectively, and a compressive strength of 80 MPa was obtained. Mineralization study in simulated body fluid (SBF) showed formation of apatite layer on the dense HA compacts indicating a good tendency of bond formation with natural bone. Cytotoxicity results showed excellent cell attachment on the HA surface. In the Appendices, 3 journal articles have been attached which describe synthesis, processing and characterization of undoped and doped PZT nanopowders. Free standing and agglomerated PZT nanopowders were synthesized by the sucrose templated method and the citrate nitrate autocombustion method. Particle size in the range

  15. Comparison of bone regeneration in alveolar bone of dogs on mineralized collagen grafts with two composition ratios of nano-hydroxyapatite and collagen

    PubMed Central

    Wang, Yan-Fu; Wang, Cheng-Yue; Wan, Peng; Wang, Shao-Gang; Wang, Xiu-Mei

    2016-01-01

    To study the effect of two composition ratios of nano-hydroxyapatite and collagen (NHAC) composites on repairing alveolar bone defect of dogs. Eighteen healthy adult dogs were randomly divided into three groups. Two kinds of the NHAC composites were prepared according to the constituent ratios of 3:7 and 5:5; immediately after extraction of the mandibular second premolars, each kind of the NHAC composite was implanted into extraction socket, respectively: Group I, nHA/Col = 3:7; Group II, nHA/Col = 5:5 and Group III, blank control group. The bone-repairing ability of the two grafts was separately analyzed by morphometric measurement, X-ray tomography examination and biomechanical analysis at 1st, 3rd and 6th month post-surgical, respectively. The NHAC composites were absorbed gradually after implanting into alveolar bone defect and were replaced by new bone. The ratios of new bone formation of Group I was significantly higher than that of Group II after 3 months (P < 0.05). The structure and bioactive performance can be improved when the ratio between the collagen and the hydroxyapatite was reasonable, and the repairing ability and effect in extraction sockets are obviously better. PMID:26816654

  16. Human Cementum Protein 1 induces expression of bone and cementum proteins by human gingival fibroblasts

    SciTech Connect

    Carmona-Rodriguez, Bruno; Alvarez-Perez, Marco Antonio; Narayanan, A. Sampath; Zeichner-David, Margarita; Reyes-Gasga, Jose; Molina-Guarneros, Juan; Garcia-Hernandez, Ana Lilia; Suarez-Franco, Jose Luis; Chavarria, Ivet Gil; Villarreal-Ramirez, Eduardo; Arzate, Higinio . E-mail: harzate@servidor.unam.mx

    2007-07-06

    We recently presented evidence showing that a human cementoblastoma-derived protein, named Cementum Protein 1 (CEMP1) may play a role as a local regulator of cementoblast differentiation and cementum-matrix mineralization. This protein was shown to be expressed by cementoblasts and progenitor cells localized in the periodontal ligament. In this study we demonstrate that transfection of CEMP1 into human gingival fibroblasts (HGF) induces mineralization and expression of bone and cementum-matrix proteins. The transfected HGF cells had higher alkaline phosphatase activity and proliferation rate and they expressed genes for alkaline phosphatase, bone sialoprotein, osteocalcin, osteopontin, the transcription factor Runx2/Cbfa1, and cementum attachment protein (CAP). They also produced biological-type hydroxyapatite. These findings indicate that the CEMP1 might participate in differentiation and mineralization of nonosteogenic cells, and that it might have a potential function in cementum and bone formation.

  17. Histologic Evaluation of Human Alveolar Sockets Treated With an Artificial Bone Substitute Material

    PubMed Central

    Wakimoto, Mari; Ueno, Takaaki; Hirata, Azumi; Iida, Seiji; Aghaloo, Tara; Moy, Peter K.

    2012-01-01

    This study involved a histologic, enzyme histologic, immunohistologic, and three-dimensional microstructure evaluating the extent of osteogenesis and repair in the human alveolar extraction socket achievable with an artificial bone substitute. After tooth extraction in 7 patients, extraction sockets were filled with Mastergraft (15% hydroxyapatite, 85% β-tricalcium phosphate complex). Radio-micrographs and histologic examinations were performed on samples obtained during dental implant placement procedure. On micro– computed tomography, new bone was observed in all collected samples, and osteogenesis was observed to have taken place around the artificial bone substitute. Histologically, active osteogenesis was found throughout the region observed. Addition of new bone around the Mastergraft was observed, and osteoblast-like cells were present. Cells that had partially invaded the artificial bone included tartrate-resistant acid phosphate–positive and CD34-positive cells. These findings indicate that the Mastergraft artificial bone induced osteogenesis in the jawbone and seemed effective for repairing bone defects. PMID:21415629

  18. Selenium-Substituted Hydroxyapatite/Biodegradable Polymer/Pamidronate Combined Scaffold for the Therapy of Bone Tumour

    PubMed Central

    Oledzka, Ewa; Sobczak, Marcin; Kolmas, Joanna; Nalecz-Jawecki, Grzegorz

    2015-01-01

    The present study evaluated a new concept of combined scaffolds as a promising bone replacement material for patients with a bone tumour or bone metastasis. The scaffolds were composed of hydroxyapatite doped with selenium ions and a biodegradable polymer (linear or branched), and contained an active substance—bisphosphonate. For this purpose, a series of biodegradable polyesters were synthesized through a ring-opening polymerization of ε-caprolactone or d,l-lactide in the presence of 2-hydroxyethyl methacrylate (HEMA) or hyperbranched 2,2-bis(hydroxymethyl)propionic acid polyester-16-hydroxyl (bis-MPA) initiators, substances often used in the synthesis of medical materials. The polymers were obtained with a high yield and a number-average molecular weight up to 45,300 (g/mol). The combined scaffolds were then manufactured by a direct compression of pre-synthesized hydroxyapatite doped with selenite or selenate ions, obtained polymer and pamidronate as a model drug. It was found that the kinetic release of the drug from the scaffolds tested in vitro under physiological conditions is strongly dependent on the physicochemical properties and average molecular weight of the polymers. Furthermore, there was good correlation with the hydrolytic biodegradation results of the scaffolds fabricated without drug. The preliminary findings suggest that the fabricated combined scaffolds could be effectively used for the sustained delivery of bioactive molecules at bone defect sites. PMID:26389884

  19. Selenium-Substituted Hydroxyapatite/Biodegradable Polymer/Pamidronate Combined Scaffold for the Therapy of Bone Tumour.

    PubMed

    Oledzka, Ewa; Sobczak, Marcin; Kolmas, Joanna; Nalecz-Jawecki, Grzegorz

    2015-01-01

    The present study evaluated a new concept of combined scaffolds as a promising bone replacement material for patients with a bone tumour or bone metastasis. The scaffolds were composed of hydroxyapatite doped with selenium ions and a biodegradable polymer (linear or branched), and contained an active substance-bisphosphonate. For this purpose, a series of biodegradable polyesters were synthesized through a ring-opening polymerization of ε-caprolactone or d,l-lactide in the presence of 2-hydroxyethyl methacrylate (HEMA) or hyperbranched 2,2-bis(hydroxymethyl)propionic acid polyester-16-hydroxyl (bis-MPA) initiators, substances often used in the synthesis of medical materials. The polymers were obtained with a high yield and a number-average molecular weight up to 45,300 (g/mol). The combined scaffolds were then manufactured by a direct compression of pre-synthesized hydroxyapatite doped with selenite or selenate ions, obtained polymer and pamidronate as a model drug. It was found that the kinetic release of the drug from the scaffolds tested in vitro under physiological conditions is strongly dependent on the physicochemical properties and average molecular weight of the polymers. Furthermore, there was good correlation with the hydrolytic biodegradation results of the scaffolds fabricated without drug. The preliminary findings suggest that the fabricated combined scaffolds could be effectively used for the sustained delivery of bioactive molecules at bone defect sites. PMID:26389884

  20. Biological assessment of a calcium silicate incorporated hydroxyapatite-gelatin nanocomposite: a comparison to decellularized bone matrix.

    PubMed

    Lee, Dong Joon; Padilla, Ricardo; Zhang, He; Hu, Wei-Shou; Ko, Ching-Chang

    2014-01-01

    Our laboratory utilized biomimicry to develop a synthetic bone scaffold based on hydroxyapatite-gelatin-calcium silicate (HGCS). Here, we evaluated the potential of HGCS scaffold in bone formation in vivo using the rat calvarial critical-sized defect (CSD). Twelve Sprague-Dawley rats were randomized to four groups: control (defect only), decellularized bone matrix (DECBM), and HGCS with and without multipotent adult progenitor cells (MAPCs). DECBM was prepared by removing all the cells using SDS and NH4OH. After 12 weeks, the CSD specimens were harvested to evaluate radiographical, histological, and histomorphometrical outcomes. The in vitro osteogenic effects of the materials were studied by focal adhesion, MTS, and alizarin red. Micro-CT analysis indicated that the DECBM and the HGCS scaffold groups developed greater radiopaque areas than the other groups. Bone regeneration, assessed using histological analysis and fluorochrome labeling, was the highest in the HGCS scaffold seeded with MAPCs. The DECBM group showed limited osteoinductivity, causing a gap between the implant and host tissue. The group grafted with HGCS+MAPCs resulting in twice as much new bone formation seems to indicate a role for effective bone regeneration. In conclusion, the novel HGCS scaffold could improve bone regeneration and is a promising carrier for stem cell-mediated bone regeneration. PMID:25054149

  1. Organic matter within crystalline aggregates of hydroxyapatite: A new substrate for stable isotopic and possibly other biogeochemical analyses of bone

    SciTech Connect

    DeNiro, M.J.; Weiner, S. )

    1988-10-01

    The portion of organic matrix in bones that is present within fused aggregates of hydroxyapatite crystals was isolated by oxidizing the rest of the bone organic matter with sodium hypochlorite. The aggregate organic matter from a variety of modern and prehistoric bones was subjected to elemental and stable C and N isotopic analysis. For comparison, collagen from modern bones and the fraction from prehistoric bones with the same solubility characteristics as collagen (referred to herein as collagen) were subjected to the same analytical procedures. Collagen and aggregate organic matter in modern bones have similar {delta}{sup 15}N values but dissimilar {delta}{sup 13}C values. The difference may be caused by the presence of non-collagenous proteins (NCPs) in the aggregate organic matter, because the NCPs have {delta}{sup 13}C values different from those of collagen from the same bone. The organic matter in aggregates is not subject to the same diagenetic processes that can alter the isotope ratios of collagen, and appears to retain an in vivo isotope signal even in cases in which that of collagen has been altered. These conclusions apply to samples that were burned prehistorically as well as to those that suffered postmortem alteration only in the depositional environment. The organic matter in aggregates represents a new substrate that should prove useful for stable isotopic studies and possibly for radiocarbon and other biogeochemical analyses of bone.

  2. Improvement in bone properties by using risedronate adsorbed hydroxyapatite novel nanoparticle based formulation in a rat model of osteoporosis.

    PubMed

    Sahana, H; Khajuria, Deepak Kumar; Razdan, Rema; Mahapatra, D Roy; Bhat, M R; Suresh, Sarasija; Rao, R Ramachandra; Mariappan, L

    2013-02-01

    A superior drug formulation capable of achieving efficient osteogenesis is in imperative demand for the treatment of osteoporosis. In the present study we investigated the potential of using novel risedronate-hydroxyapatite (HA) nanoparticle based formulation in an animal model of established osteoporosis. Nanoparticles of HA loaded with risedronate (NHLR) of various sizes (80-130 nm) were generated for bone targeted drug delivery. Three months after ovariectomy, 36 ovariectomized (OVX) rats were divided into 6 equal groups and treated with various doses of NHLR (500, 350 and 250 microg/kg intravenous single dose) and sodium risedronate (500 microg/kg, intravenous single dose). Untreated OVX and sham OVX served as controls. One month after drug administration, the left tibia and femur were tested for bone mechanical properties and histology, respectively. In the right femur, bone density was measured by method based on Archimedes principle and bone porosity analyses were performed using X-ray imaging. NHLR (250 microg/kg) showed a significant increase in bone density and reduced bone porosity when compared with OVX control. Moreover, NHLR (250 microg/kg) significantly increased bone mechanical properties and bone quality when compared with OVX control. The results strongly suggest that the NHLR, which is a novel nanoparticle based formulation, has a therapeutic advantage over risedronate sodium monotherapy for the treatment of osteoporosis in a rat model of postmenopausal osteoporosis. PMID:23627045

  3. Cortical bone growth and maturational changes in dwarf rats induced by recombinant human growth hormone

    NASA Technical Reports Server (NTRS)

    Martinez, D. A.; Orth, M. W.; Carr, K. E.; Vanderby, R. Jr; Vailas, A. C.

    1996-01-01

    The growth hormone (GH)-deficient dwarf rat was used to investigate recombinant human (rh) GH-induced bone formation and to determine whether rhGH facilitates simultaneous increases in bone formation and bone maturation during rapid growth. Twenty dwarf rats, 37 days of age, were randomly assigned to dwarf plus rhGH (GH; n = 10) and dwarf plus vehicle (n = 10) groups. The GH group received 1.25 mg rhGH/kg body wt two times daily for 14 days. Biochemical, morphological, and X-ray diffraction measurements were performed on the femur middiaphysis. rhGH stimulated new bone growth in the GH group, as demonstrated by significant increases (P < 0.05) in longitudinal bone length (6%), middiaphyseal cross-sectional area (20%), and the amount of newly accreted bone collagen (28%) in the total pool of middiaphyseal bone collagen. Cortical bone density, mean hydroxyapatite crystal size, and the calcium and collagen contents (microgram/mm3) were significantly smaller in the GH group (P < 0.05). Our findings suggest that the processes regulating new collagen accretion, bone collagen maturation, and mean hydroxyapatite crystal size may be independently regulated during rapid growth.

  4. Graphene oxide nanoflakes incorporated gelatin-hydroxyapatite scaffolds enhance osteogenic differentiation of human mesenchymal stem cells

    NASA Astrophysics Data System (ADS)

    Nair, Manitha; Nancy, D.; Krishnan, Amit G.; Anjusree, G. S.; Vadukumpully, Sajini; Nair, Shantikumar V.

    2015-04-01

    In this study, graphene oxide (GO) nanoflakes (0.5 and 1 wt%) were incorporated into a gelatin-hydroxyapatite (GHA) matrix through a freeze drying technique and its effect to enhance mechanical strength and osteogenic differentiation was studied. The GHA matrix with GO demonstrated less brittleness in comparison to GHA scaffolds. There was no significant difference in mechanical strength between GOGHA0.5 and GOGHA1.0 scaffolds. When the scaffolds were immersed in phosphate buffered saline (to mimic physiologic condition) for 60 days, around 50-60% of GO was released in sustained and linear manner and the concentration was within the toxicity limit as reported earlier. Further, GOGHA0.5 scaffolds were continued for cell culture experiments, wherein the scaffold induced osteogenic differentiation of human adipose derived mesenchymal stem cells without providing supplements like dexamethasone, L-ascorbic acid and β glycerophosphate in the medium. The level of osteogenic differentiation of stem cells was comparable to those cultured on GHA scaffolds with osteogenic supplements. Thus biocompatible, biodegradable and porous GO reinforced gelatin-HA 3D scaffolds may serve as a suitable candidate in promoting bone regeneration in orthopaedics.

  5. Poly(l-Lactic Acid)/Gelatin Fibrous Scaffold Loaded with Simvastatin/Beta-Cyclodextrin-Modified Hydroxyapatite Inclusion Complex for Bone Tissue Regeneration.

    PubMed

    Lee, Jung Bok; Kim, Ji Eun; Balikov, Daniel A; Bae, Min Soo; Heo, Dong Nyoung; Lee, Donghyun; Rim, Hyun Joon; Lee, Deok-Won; Sung, Hak-Joon; Kwon, Il Keun

    2016-07-01

    Recently, the application of nanostructured materials in the field of tissue engineering has garnered attention to mediate treatment and regeneration of bone defects. In this study, poly(l-lactic acid) (PLLA)/gelatin (PG) fibrous scaffolds are fabricated and β-cyclodextrin (βCD) grafted nano-hydroxyapatite (HAp) is coated onto the fibrous scaffold surface via an interaction between βCD and adamantane. Simvastatin (SIM), which is known to promote osteoblast viability and differentiation, is loaded into the remaining βCD. The specimen morphologies are characterized by scanning electron microscopy. The release profile of SIM from the drug loaded scaffold is also evaluated. In vitro proliferation and osteogenic differentiation of human adipose derived stem cells on SIM/HAp coated PG composite scaffolds is characterized by alkaline phosphatase (ALP) activity, mineralization (Alizarin Red S staining), and real time Polymerase chain reaction (PCR). The scaffolds are then implanted into rabbit calvarial defects and analyzed by microcomputed tomography for bone formation after four and eight weeks. These results demonstrate that SIM loaded PLLA/gelatin/HAp-(βCD) scaffolds promote significantly higher ALP activity, mineralization, osteogenic gene expression, and bone regeneration than control scaffolds. This suggests the potential application of this material toward bone tissue engineering. PMID:26996294

  6. Biocompatibility and bone-repairing effects: comparison between porous poly-lactic-co-glycolic acid and nano-hydroxyapatite/poly(lactic acid) scaffolds.

    PubMed

    Zong, Chen; Qian, Xiaodan; Tang, Zihua; Hu, Qinghong; Chen, Jiarong; Gao, Changyou; Tang, Ruikang; Tong, Xiangmin; Wang, Jinfu

    2014-06-01

    Copolymer composite scaffolds and bioceramic/polymer composite scaffolds are two representative forms of composite scaffolds used for bone tissue engineering. Studies to compare biocompatibility and bone-repairing effects between these two scaffolds are significant for selecting or improving the scaffold for clinical application. We prepared two porous scaffolds comprising poly-lactic-acid/poly-glycolic-acid (PLGA) and poly-lactic-acid/nano-hydroxyapatite (nHAP/PLA) respectively, and examined their biocompatibility with human bone marrow-derived mesenchymal stem cells (hMSCs) through evaluating adhesion, proliferation and osteogenic differentiation potentials of hMSCs in the scaffold. Then, the PLGA scaffold with hMSCs (PM construct) and the nHAP/PLA scaffold with hMSCs (HPM construct) were transplanted into the rat calvarial defect areas to compare their effects on the bone reconstruction. The results showed that the nHAP/PLA scaffold was in favor of adhesion, matrix deposition and osteogenic differentiation of hMSCs. For in vivo transplantation, both HPM and PM constructs led to mineralization and osteogenesis in the defect area of rat. However, the area grafted with PM construct showed a better formation of mature bone than that with HPM construct. In addition, the evaluation of in vitro and in vivo degradation indicated that the degradation rate of nHAP/PLA scaffold was much lower than that of PLGA scaffold. It is inferred that the lower degradation of nHAP/PLA scaffold should result in its inferior bone reconstruction in rat calvaria. Therefore, the preparation of an ideal composite scaffold for bone tissue engineering should be taken into account of the balance between its biocompatibility, degradation rate, osteoconductivity and mechanical property. PMID:24749403

  7. Effect of glycerol-L-lactide coating polymer on bone ingrowth of bFGF-coated hydroxyapatite implants.

    PubMed

    Alt, Volker; Pfefferle, Hans-Joachim; Kreuter, Jörg; Stahl, Jens-Peter; Pavlidis, Theodoros; Meyer, Christof; Mockwitz, Jürgen; Wenisch, Sabine; Schnettler, Reinhard

    2004-09-14

    Basic fibroblast growth factor (bFGF)-coated hydroxyapatite (HA) cylinders showed good bony incorporation in a previously conducted animal study. However, some cylinders exhibited focal inhomogeneous bone ingrowth. The purpose of the current study was to test whether glycerol-L-lactide polymer coating could improve release properties and bone incorporation of bFGF-coated HA implants. bFGF-coated HA cylinders with or without coating polymer were investigated for in vitro release of bFGF by an immuno-ligand-assay and also for bone ingrowth in miniature pigs after 42 and 84 days. Release from bFGF polymer composites was lower for the first 3 days compared to the other group but was more homogenous and detectable amounts were still found after 20 days. There was significant delay in bone ingrowth of the polymer implants in which even after 84 days bone ingrowth was not completed, whereas in the other group incorporation after 42 days occurred. Detailed histology revealed filling of the HA pores with the polymer, making ingrowth of the surrounding host bone impossible. Only after 84 days starting resorption of the polymer accompanied by bone ingrowth was found. The current study showed that glycerol-L-lactide is not suitable for coating of HA implants due to polymer induced "locking" of HA pores. PMID:15342184

  8. Preparation and evaluation of an Arg-Gly-Asp-modified chitosan/hydroxyapatite scaffold for application in bone tissue engineering

    PubMed Central

    CHEN, LIN; LI, BAOLIN; XIAO, XIAO; MENG, QINGGANG; LI, WEI; YU, QIAN; BI, JIAQI; CHENG, YONG; QU, ZHIWEI

    2015-01-01

    Bone tissue engineering has become a promising method for the repair of bone defects, and the production of a scaffold with high cell affinity and osseointegrative properties is crucial for successful bone substitute. Chitosan (CS)/hydroxyapatite (HA) composite was prepared by in situ compositing combined with lyophilization, and further modified by arginine-glycine-aspartic acid (RGD) via physical adsorption. In order to evaluate the cell adhesion rate, viability, morphology, and alkaline phosphatase (ALP) activity, the RGD-CS/HA scaffold was seeded with bone marrow stromal cells (BMSCs). The osseointegrative properties of the RGD-CS/HA scaffold were evaluated by in vivo heterotopic ossification and in vivo bone defect repair. After 4 h culture with the RGD-CS/HA scaffold, the adhesion rate of the BMSCs was 80.7%. After 3 days, BMSCs were fusiform in shape and evenly distributed on the RGD-CS/HA scaffold. Formation of extracellular matrix and numerous cell-cell interactions were observed after 48 h of culture, with an ALP content of 0.006±0.0008 U/l/ng. Furthermore, the osseointegrative ability and biomechanical properties of the RGD-CS/HA scaffold were comparable to that of normal bone tissue. The biocompatibility, cytocompatibility, histocompatibility and osseointegrative properties of the RGD-CS/HA scaffold support its use in bone tissue engineering applications. PMID:26459053

  9. The biocompatibility of carbon hydroxyapatite/β-glucan composite for bone tissue engineering studied with Raman and FTIR spectroscopic imaging.

    PubMed

    Sroka-Bartnicka, Anna; Kimber, James A; Borkowski, Leszek; Pawlowska, Marta; Polkowska, Izabela; Kalisz, Grzegorz; Belcarz, Anna; Jozwiak, Krzysztof; Ginalska, Grazyna; Kazarian, Sergei G

    2015-10-01

    The spectroscopic approaches of FTIR imaging and Raman mapping were applied to the characterisation of a new carbon hydroxyapatite/β-glucan composite developed for bone tissue engineering. The composite is an artificial bone material with an apatite-forming ability for the bone repair process. Rabbit bone samples were tested with an implanted bioactive material for a period of several months. Using spectroscopic and chemometric methods, we were able to determine the presence of amides and phosphates and the distribution of lipid-rich domains in the bone tissue, providing an assessment of the composite's bioactivity. Samples were also imaged in transmission using an infrared microscope combined with a focal plane array detector. CaF2 lenses were also used on the infrared microscope to improve spectral quality by reducing scattering artefacts, improving chemometric analysis. The presence of collagen and lipids at the bone/composite interface confirmed biocompatibility and demonstrate the suitability of FTIR microscopic imaging with lenses in studying these samples. It confirmed that the composite is a very good background for collagen growth and increases collagen maturity with the time of the bone growth process. The results indicate the bioactive and biocompatible properties of this composite and demonstrate how Raman and FTIR spectroscopic imaging have been used as an effective tool for tissue characterisation. PMID:26277184

  10. Preparation and evaluation of an Arg-Gly-Asp-modified chitosan/hydroxyapatite scaffold for application in bone tissue engineering.

    PubMed

    Chen, Lin; Li, Baolin; Xiao, Xiao; Meng, Qinggang; Li, Wei; Yu, Qian; Bi, Jiaqi; Cheng, Yong; Qu, Zhiwei

    2015-11-01

    Bone tissue engineering has become a promising method for the repair of bone defects, and the production of a scaffold with high cell affinity and osseointegrative properties is crucial for successful bone substitute. Chitosan (CS)/hydroxyapatite (HA) composite was prepared by in situ compositing combined with lyophilization, and further modified by arginine‑glycine‑aspartic acid (RGD) via physical adsorption. In order to evaluate the cell adhesion rate, viability, morphology, and alkaline phosphatase (ALP) activity, the RGD‑CS/HA scaffold was seeded with bone marrow stromal cells (BMSCs). The osseointegrative properties of the RGD‑CS/HA scaffold were evaluated by in vivo heterotopic ossification and in vivo bone defect repair. After 4 h culture with the RGD‑CS/HA scaffold, the adhesion rate of the BMSCs was 80.7%. After 3 days, BMSCs were fusiform in shape and evenly distributed on the RGD‑CS/HA scaffold. Formation of extracellular matrix and numerous cell‑cell interactions were observed after 48 h of culture, with an ALP content of 0.006 ± 0.0008 U/l/ng. Furthermore, the osseointegrative ability and biomechanical properties of the RGD‑CS/HA scaffold were comparable to that of normal bone tissue. The biocompatibility, cytocompatibility, histocompatibility and osseointegrative properties of the RGD‑CS/HA scaffold support its use in bone tissue engineering applications. PMID:26459053

  11. Comparative Study of Bone Repair Using Porous Hydroxyapatite/ β-Tricalcium Phosphate and Xenograft Scaffold in Rabbits with Tibia Defect

    PubMed Central

    Bagher, Zohreh; Rajaei, Farzad; Shokrgozar, Mohammadali

    2012-01-01

    Background: Bone tissue engineering requires materials that are biocompatible, mechanically suited for bone function, integrated with the host skeleton, and support osteoinduction of the implanted cells for new bone formation. The aim of this study was to compare the osteogenic potential of xenograft with hydroxyapatite/β- tricalcium phosphate (HA/β-TCP) scaffold. Methods: New Zealand rabbits (n = 9) were divided into 3 groups. Osteoblast cells were originally isolated from rabbit iliac crest and cultured in DMEM/F12. After creating a critical-sized defect (2 × 3 cm) in rabbit tibia bone, the defect was filled with an implant of HA/TCP with osteoblasts and xenograft in the hole of left (as control) and right tibia, respectively. The new bone formation and the development of bone union within the defect were evaluated by x-ray images and eosine and hematoxylin staining at 4, 8, and 12 weeks post-operation. Results: The bone partially formed in both groups was filled with osteoblast cultured on porous implants at 4 weeks. Over time, progressive bone regeneration was observed inside the pores. Moreover, a progressive vascular ingrowth and progressive integration with the host bone were obvious in xenograft when compared to HA/β-TCP. A good integration between the xenograft implants and the bone was observed radiographically and confirmed by histological section. Conclusion: The result showed that the bone defect can be repaired using both synthetic and xenograft implants. However, the xenograft showed a better osteointegration as compared to HA/β-TCP scaffold. PMID:22562028

  12. Strontium-Doped Calcium Phosphate and Hydroxyapatite Granules Promote Different Inflammatory and Bone Remodelling Responses in Normal and Ovariectomised Rats

    PubMed Central

    Xia, Wei; Emanuelsson, Lena; Norlindh, Birgitta; Omar, Omar; Thomsen, Peter

    2013-01-01

    The healing of bone defects may be hindered by systemic conditions such as osteoporosis. Calcium phosphates, with or without ion substitutions, may provide advantages for bone augmentation. However, the mechanism of bone formation with these materials is unclear. The aim of this study was to evaluate the healing process in bone defects implanted with hydroxyapatite (HA) or strontium-doped calcium phosphate (SCP) granules, in non-ovariectomised (non-OVX) and ovariectomised (OVX) rats. After 0 (baseline), six and 28d, bone samples were harvested for gene expression analysis, histology and histomorphometry. Tumour necrosis factor-α (TNF-α), at six days, was higher in the HA, in non-OVX and OVX, whereas interleukin-6 (IL-6), at six and 28d, was higher in SCP, but only in non-OVX. Both materials produced a similar expression of the receptor activator of nuclear factor kappa-B ligand (RANKL). Higher expression of osteoclastic markers, calcitonin receptor (CR) and cathepsin K (CatK), were detected in the HA group, irrespective of non-OVX or OVX. The overall bone formation was comparable between HA and SCP, but with topological differences. The bone area was higher in the defect centre of the HA group, mainly in the OVX, and in the defect periphery of the SCP group, in both non-OVX and OVX. It is concluded that HA and SCP granules result in comparable bone formation in trabecular bone defects. As judged by gene expression and histological analyses, the two materials induced different inflammatory and bone remodelling responses. The modulatory effects are associated with differences in the spatial distribution of the newly formed bone. PMID:24376855

  13. Strontium-doped calcium phosphate and hydroxyapatite granules promote different inflammatory and bone remodelling responses in normal and ovariectomised rats.

    PubMed

    Cardemil, Carina; Elgali, Ibrahim; Xia, Wei; Emanuelsson, Lena; Norlindh, Birgitta; Omar, Omar; Thomsen, Peter

    2013-01-01

    The healing of bone defects may be hindered by systemic conditions such as osteoporosis. Calcium phosphates, with or without ion substitutions, may provide advantages for bone augmentation. However, the mechanism of bone formation with these materials is unclear. The aim of this study was to evaluate the healing process in bone defects implanted with hydroxyapatite (HA) or strontium-doped calcium phosphate (SCP) granules, in non-ovariectomised (non-OVX) and ovariectomised (OVX) rats. After 0 (baseline), six and 28d, bone samples were harvested for gene expression analysis, histology and histomorphometry. Tumour necrosis factor-α (TNF-α), at six days, was higher in the HA, in non-OVX and OVX, whereas interleukin-6 (IL-6), at six and 28d, was higher in SCP, but only in non-OVX. Both materials produced a similar expression of the receptor activator of nuclear factor kappa-B ligand (RANKL). Higher expression of osteoclastic markers, calcitonin receptor (CR) and cathepsin K (CatK), were detected in the HA group, irrespective of non-OVX or OVX. The overall bone formation was comparable between HA and SCP, but with topological differences. The bone area was higher in the defect centre of the HA group, mainly in the OVX, and in the defect periphery of the SCP group, in both non-OVX and OVX. It is concluded that HA and SCP granules result in comparable bone formation in trabecular bone defects. As judged by gene expression and histological analyses, the two materials induced different inflammatory and bone remodelling responses. The modulatory effects are associated with differences in the spatial distribution of the newly formed bone. PMID:24376855

  14. Fabrication, Properties and Applications of Dense Hydroxyapatite: A Review

    PubMed Central

    Prakasam, Mythili; Locs, Janis; Salma-Ancane, Kristine; Loca, Dagnija; Largeteau, Alain; Berzina-Cimdina, Liga

    2015-01-01

    In the last five decades, there have been vast advances in the field of biomaterials, including ceramics, glasses, glass-ceramics and metal alloys. Dense and porous ceramics have been widely used for various biomedical applications. Current applications of bioceramics include bone grafts, spinal fusion, bone repairs, bone fillers, maxillofacial reconstruction, etc. Amongst the various calcium phosphate compositions, hydroxyapatite, which has a composition similar to human bone, has attracted wide interest. Much emphasis is given to tissue engineering, both in porous and dense ceramic forms. The current review focusses on the various applications of dense hydroxyapatite and other dense biomaterials on the aspects of transparency and the mechanical and electrical behavior. Prospective future applications, established along the aforesaid applications of hydroxyapatite, appear to be promising regarding bone bonding, advanced medical treatment methods, improvement of the mechanical strength of artificial bone grafts and better in vitro/in vivo methodologies to afford more particular outcomes. PMID:26703750

  15. Fabrication, Properties and Applications of Dense Hydroxyapatite: A Review.

    PubMed

    Prakasam, Mythili; Locs, Janis; Salma-Ancane, Kristine; Loca, Dagnija; Largeteau, Alain; Berzina-Cimdina, Liga

    2015-01-01

    In the last five decades, there have been vast advances in the field of biomaterials, including ceramics, glasses, glass-ceramics and metal alloys. Dense and porous ceramics have been widely used for various biomedical applications. Current applications of bioceramics include bone grafts, spinal fusion, bone repairs, bone fillers, maxillofacial reconstruction, etc. Amongst the various calcium phosphate compositions, hydroxyapatite, which has a composition similar to human bone, has attracted wide interest. Much emphasis is given to tissue engineering, both in porous and dense ceramic forms. The current review focusses on the various applications of dense hydroxyapatite and other dense biomaterials on the aspects of transparency and the mechanical and electrical behavior. Prospective future applications, established along the aforesaid applications of hydroxyapatite, appear to be promising regarding bone bonding, advanced medical treatment methods, improvement of the mechanical strength of artificial bone grafts and better in vitro/in vivo methodologies to afford more particular outcomes. PMID:26703750

  16. Carbon nanotube-reinforced hydroxyapatite composite and their interaction with human osteoblast in vitro.

    PubMed

    Khalid, P; Hussain, M A; Rekha, P D; Arun, A B

    2015-05-01

    As a bone mineral component, hydroxyapatite (HA) has been an attractive bioceramic for the reconstruction of hard tissues. However, its poor mechanical properties, including low fracture toughness and tensile strength, have been a substantial challenge to the application of HA for the replacement of load-bearing and/or large bone defects. In this study, HA is reinforced with high-purity and well-functionalized multiwalled carbon nanotubes (MWCNTs; >99 wt%) having an average diameter of 15 nm and length from 10 to 20 μm. The cellular response of these functionalized CNTs and its composites were examined in human osteoblast sarcoma cell lines. Calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) and diammonium hydrogen phosphate ((NH4)2HPO4) were used to synthesize HA in situ. MWCNTs were functionalized by heating at 100°C in 3:1 ratio of sulfuric acid and nitric acid for 60 min with stirring and dispersed in sodium dodecyl benzene sulfonate by sonication. HA particles were produced in MWCNTs solution by adding Ca(NO3)2·4H2O and (NH4)2HPO4 under vigorously stirring conditions. The composite was dried and washed in distilled water followed by heat treatment at 250°C to obtain CNT-HA powder. Physiochemical characterization of the composite material was carried out using Fourier transform infrared spectroscopy, field-emission scanning electron microscopy, energy-dispersive X-ray spectrometer, and X-ray diffractometer. Furthermore, this study investigates the cytotoxic effects of functionalized-MWCNTs (f-MWCNTs) and its composites with HA in human osteoblast sarcoma cell lines. Human osteoblast cells were exposed with different concentrations of f-MWCNTs and its composite with HA. The interactions of f-MWCNT and MWCNT-HA composites were analyzed by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The results indicate no detrimental effect on survival or mitochondrial activity of the osteoblast cells. Cell viability decreased with an increase in CNT

  17. Open-Porous Hydroxyapatite Scaffolds for Three-Dimensional Culture of Human Adult Liver Cells

    PubMed Central

    Schmelzer, Eva; Over, Patrick; Nettleship, Ian; Gerlach, Joerg C.

    2016-01-01

    Liver cell culture within three-dimensional structures provides an improved culture system for various applications in basic research, pharmacological screening, and implantable or extracorporeal liver support. Biodegradable calcium-based scaffolds in such systems could enhance liver cell functionality by providing endothelial and hepatic cell support through locally elevated calcium levels, increased surface area for cell attachment, and allowing three-dimensional tissue restructuring. Open-porous hydroxyapatite scaffolds were fabricated and seeded with primary adult human liver cells, which were embedded within or without gels of extracellular matrix protein collagen-1 or hyaluronan. Metabolic functions were assessed after 5, 15, and 28 days. Longer-term cultures exhibited highest cell numbers and liver specific gene expression when cultured on hydroxyapatite scaffolds in collagen-1. Endothelial gene expression was induced in cells cultured on scaffolds without extracellular matrix proteins. Hydroxyapatite induced gene expression for cytokeratin-19 when cells were cultured in collagen-1 gel while culture in hyaluronan increased cytokeratin-19 gene expression independent of the use of scaffold in long-term culture. The implementation of hydroxyapatite composites with extracellular matrices affected liver cell cultures and cell differentiation depending on the type of matrix protein and the presence of a scaffold. The hydroxyapatite scaffolds enable scale-up of hepatic three-dimensional culture models for regenerative medicine applications. PMID:27403430

  18. Hydroxyapatite from fish scale for potential use as bone scaffold or regenerative material.

    PubMed

    Pon-On, Weeraphat; Suntornsaratoon, Panan; Charoenphandhu, Narattaphol; Thongbunchoo, Jirawan; Krishnamra, Nateetip; Tang, I Ming

    2016-05-01

    The present paper studies the physico-chemical, bioactivity and biological properties of hydroxyapatite (HA) which is derived from fish scale (FS) (FSHA) and compares them with those of synthesized HA (sHA) obtained by co-precipitation from chemical solution as a standard. The analysis shows that the FSHA is composed of flat-plate nanocrystal with a narrow width size of about 15-20 nm and having a range of 100 nm in length and that the calcium phosphate ratio (Ca/P) is 2.01 (Ca-rich CaP). Whereas, synthesized HA consists of sub-micron HA particle having a Ca/P ratio of 1.65. Bioactivity test shows that the FSHA forms more new apatite than does the sHA after being incubated in simulated body fluid (SBF) for 7 days. Moreover, the biocompatibility study shows a higher osteoblast like cell adhesion on the FSHA surface than on the sHA substrate after 3 days of culturing. Our results also show the shape of the osteoblast cells on the FSHA changes from being a rounded shape to being a flattened shape reflecting its spreading behavior on this surface. MTT assay and ALP analysis show significant increases in the proliferation and activity of osteoblasts over the FSHA scaffold after 5 days of culturing as compared to those covering the sHA substrates. These results confirm that the bio-materials derived from fish scale (FSHA) are biologically better than the chemically synthesized HA and have the potential for use as a bone scaffold or as regenerative materials. PMID:26952413

  19. Mechanical and cytotoxicity evaluation of nanostructured hydroxyapatite-bredigite scaffolds for bone regeneration.

    PubMed

    Eilbagi, Marjan; Emadi, Rahmatollah; Raeissi, Keyvan; Kharaziha, Mahshid; Valiani, Ali

    2016-11-01

    Despite the attractive characteristics of three-dimensional pure hydroxyapatite (HA) scaffolds, due to their weak mechanical properties, researches have focused on the development of composite scaffolds via introducing suitable secondary components. The aim of this study was to develop, for the first time, three-dimensional HA-bredigite (Ca7MgSi4O16) scaffolds containing various amounts of bredigite nanopowder (0, 5, 10 and 15wt.%) using space holder technique. Transmission electron microscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction spectroscopy were applied in order to study the morphology, fracture surface and phase compositions of nanopowders and scaffolds. Furthermore, the effects of scaffold composition on the mechanical properties, bioactivity, biodegradability, and cytotoxicity were also evaluated. Results showed that the composite scaffolds with average pore size in the range of 220-310μm, appearance porosity of 63.1-75.9% and appearance density of 1.1±0.04g/cm(3) were successfully developed, depending on bredigite content. Indeed, the micropore size of the scaffolds reduced with increasing bredigite content confirming that the sinterability of the scaffolds was improved. Furthermore, the compression strength and modulus of the scaffolds significantly enhanced via incorporation of bredigite content from 0 to 15wt.%. The composite scaffolds revealed superior bioactivity and biodegradability with increasing bredigite content. Moreover, MTT assay confirmed that HA-15wt.% bredigite scaffold significantly promoted cell proliferation compared to tissue culture plate (control) and HA scaffold. Based on these results, three-dimensional HA-bredigite scaffolds could be promising replacements for HA scaffolds in bone regeneration. PMID:27524060

  20. Magnetic resonance functional nano-hydroxyapatite incorporated poly(caprolactone) composite scaffolds for in situ monitoring of bone tissue regeneration by MRI.

    PubMed

    Ganesh, Nitya; Ashokan, Anusha; Rajeshkannan, Ramiah; Chennazhi, Krishnaprasad; Koyakutty, Manzoor; Nair, Shantikumar V

    2014-10-01

    In this study, we have reported the incorporation of a multi-modal contrast agent based on hydroxyapatite nanocrystals, within a poly(caprolactone)(PCL) nanofibrous scaffold by electrospinning. The multifunctional hydroxyapatite nanoparticles (MF-nHAp) showed simultaneous contrast enhancement for three major molecular imaging techniques. In this article, the magnetic resonance (MR) contrast enhancement ability of the MF-nHAp was exploited for the purpose of potentially monitoring as well as for influencing tissue regeneration. These MF-nHAp containing PCL scaffolds were engineered in order to enhance the osteogenic potential as well as its MR functionality for their application in bone tissue engineering. The nano-composite scaffolds along with pristine PCL were evaluated physico-chemically and biologically in vitro, in the presence of human mesenchymal stem cells (hMSCs). The incorporation of 30-40 nm sized MF-nHAp within the nanofibers showed a substantial increase in scaffold strength, protein adsorption, proliferation, and osteogenic differentiation of hMSCs along with enhanced MR functionality. This preliminary study was performed to eventually exploit the MR contrast imaging capability of MF-nHAp in nanofibrous scaffolds for real-time imaging of the changes in the tissue engineered construct. PMID:24785187

  1. Magnetic Resonance Functional Nano-Hydroxyapatite Incorporated Poly(Caprolactone) Composite Scaffolds for In Situ Monitoring of Bone Tissue Regeneration by MRI

    PubMed Central

    Ganesh, Nitya; Ashokan, Anusha; Rajeshkannan, Ramiah; Chennazhi, Krishnaprasad; Koyakutty, Manzoor

    2014-01-01

    In this study, we have reported the incorporation of a multi-modal contrast agent based on hydroxyapatite nanocrystals, within a poly(caprolactone)(PCL) nanofibrous scaffold by electrospinning. The multifunctional hydroxyapatite nanoparticles (MF-nHAp) showed simultaneous contrast enhancement for three major molecular imaging techniques. In this article, the magnetic resonance (MR) contrast enhancement ability of the MF-nHAp was exploited for the purpose of potentially monitoring as well as for influencing tissue regeneration. These MF-nHAp containing PCL scaffolds were engineered in order to enhance the osteogenic potential as well as its MR functionality for their application in bone tissue engineering. The nano-composite scaffolds along with pristine PCL were evaluated physico-chemically and biologically in vitro, in the presence of human mesenchymal stem cells (hMSCs). The incorporation of 30–40 nm sized MF-nHAp within the nanofibers showed a substantial increase in scaffold strength, protein adsorption, proliferation, and osteogenic differentiation of hMSCs along with enhanced MR functionality. This preliminary study was performed to eventually exploit the MR contrast imaging capability of MF-nHAp in nanofibrous scaffolds for real-time imaging of the changes in the tissue engineered construct. PMID:24785187

  2. In Vitro Corrosion and Cytocompatibility Properties of Nano-Whisker Hydroxyapatite Coating on Magnesium Alloy for Bone Tissue Engineering Applications

    PubMed Central

    Yang, Huawei; Yan, Xueyu; Ling, Min; Xiong, Zuquan; Ou, Caiwen; Lu, Wei

    2015-01-01

    We report here the successful fabrication of nano-whisker hydroxyapatite (nHA) coatings on Mg alloy by using a simple one-step hydrothermal process in aqueous solution. The nHA coating shows uniform structure and high crystallinity. Results indicate that nHA coating is promising for improving the in vitro corrosion and cytocompatibility properties of Mg-based implants and devices for bone tissue engineering. In addition, the simple hydrothermal deposition method used in the current study is also applicable to substrates with complex shapes or surface geometries. PMID:25789500

  3. Evaluation of an injectable bone substitute (betaTCP/hydroxyapatite/hydroxy-propyl-methyl-cellulose) in severely osteopenic and aged rats.

    PubMed

    Blouin, S; Moreau, M F; Weiss, P; Daculsi, G; Baslé, M F; Chappard, D

    2006-09-01

    The use of injectable biomaterials is of interest in osteoporotic patients to locally restore bone mass in sites at risk of fracture. An injectable bone substitute (IBS1 made of betaTCP/hydroxyapatite as a calcium phosphate substitute and hydroxy-propyl-methyl-cellulose as a polymer carrier) was used in a severely osteopenic rat model obtained by combining orchidectomy (ORX) and disuse (paralysis induced by botulinum toxin - BTX). Fifty-six aged male rats were randomized into three groups: 18 were SHAM operated; 38 were ORX and BTX injected in the right hindlimb; they constituted the OP (osteoporotic) group. One month after ORX-BTX surgery, 20 of these OP rats received a IBS1 injection in the right femur (OP-IBS1 rats). Animals were studied at the time of IBS1 injection 1 month post ORX-BTX (M1), 1 month (M2) and 2 months (M3) after IBS1 injection. Bone mass (BV/TV) and microarchitectural parameters were measured by microCT. BV/TV was decreased after ORX-BTX; ORX and BTX had cumulative effects on bone loss (differences maximized on the right femur). BV/TV (combining the volume of both bone and material in OP-IBS1 rats) was elevated at M1 but decreased at M2. Marked bone formation was found onto the biomaterial granules but bone had a woven texture. A marked increase in the number of nonosteoclastic TRAcP+ cells was found in the implanted area. IBS1 induced new bone formation shortly after implantation but both IBS1 and woven bone were resorbed without inducing lamellar bone. Biomaterial trials must be conducted with long-term implantation periods, in aged osteoporotic animals. PMID:16739169

  4. Synthesis of antimicrobial monophase silver-doped hydroxyapatite nanopowders for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Stanić, Vojislav; Janaćković, Djordje; Dimitrijević, Suzana; Tanasković, Sladjana B.; Mitrić, Miodrag; Pavlović, Mirjana S.; Krstić, Aleksandra; Jovanović, Dragoljub; Raičević, Slavica

    2011-02-01

    Monophase silver-doped hydroxyapatite (AgxCa10-x(PO4)6(OH)2; 0.002 ≤ x ≤ 0.04) nanoparticles were prepared using a neutralization method and investigated with respect to potential medical applications. This method consists of dissolving Ag2O in solution of H3PO4, and the slow addition to suspension of Ca(OH)2 was applied for the purpose of homogenous distribution of silver ions. Characterization studies from XRD, TEM and FTIR spectra showed that obtained crystals are monophase hydroxyapatites and that particles of all samples are of nano size, with average length of 70 nm and about 15-25 nm in diameter. Antimicrobial studies have demonstrated that all silver-doped hydroxyapatite samples exhibit excellent antimicrobial activity in vitro against the following pathogens: Staphylococcus aureus, Escherichia coli and Candida albicans. The hydroxyapatite sample with the highest content of silver has shown the highest antimicrobial activity; killed all cells of E. coli and brought to more than 99% reduction in viable counts of S. aureus and C. albicans. The atomic force microscopic studies illustrate that silver-doped hydroxyapatite sample causes considerable morphological changes of microorganism cells which might be the cause of cells' death. Hemolysis ratios of the silver-doped hydroxyapatite samples were below 3%, indicating good blood compatibility and that are promising as biomaterials.

  5. Degradation and biocompatibility of porous nano-hydroxyapatite/polyurethane composite scaffold for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Dong, Zhihong; Li, Yubao; Zou, Qin

    2009-04-01

    Porous scaffold containing 30 wt% nano-hydroxyapatite (n-HA) and 70 wt% polyurethane (PU) from castor oil was prepared by a foaming method and investigated by X-ray diffraction (XRD), Fourier transform infrared absorption (FTIR), scanning electron microscopy (SEM) techniques. The results show that n-HA particles disperse homogeneously in the PU matrix. The porous scaffold has not only macropores of 100-800 μm in size but also a lot of micropores on the walls of macropores. The porosity and compressive strength of scaffold are 80% and 271 kPa, respectively. After soaking in simulated body fluid (SBF), hydrolysis and deposition partly occur on the scaffold. The biological evaluation in vitro and in vivo shows that the n-HA/PU scaffold is non-cytotoxic and degradable. The porous structure provides a good microenvironment for cell adherence, growth and proliferation. The n-HA/PU composite scaffold can be satisfied with the basic requirement for tissue engineering, and has the potential to be applied in repair and substitute of human menisci of the knee-joint and articular cartilage.

  6. Efficacy of nano-hydroxyapatite prepared by an aqueous solution combustion technique in healing bone defects of goat

    PubMed Central

    Nandi, Samit Kumar; Ghosh, Samir Kumar; De, Dipak Kumar; Basu, Debabrata

    2008-01-01

    The present study was undertaken to evaluate porous hydroxyapatite (HAp), the powder of which was prepared by a novel aqueous solution combustion technique, as a bone substitute in healing bone defects in vivo, as assessed by radiologic and histopathologic methods, oxytetracycline labeling, and angiogenic features in Bengal goat. Bone defects were created in the diaphysis of the radius and either not filled (group I) or filled with a HAp strut (group II). The radiologic study in group II showed the presence of unabsorbed implants which acted as a scaffold for new bone growth across the defect, and the quality of healing of the bone defect was almost indistinguishable from the control group, in which the defect was more or less similar, although the newly formed bony tissue was more organized when HAp was used. Histologic methods showed complete normal ossification with development of Haversian canals and well-defined osteoblasts at the periphery in group II, whereas the control group had moderate fibro-collagenization and an adequate amount of marrow material, fat cells, and blood vessels. An oxytetracycline labeling study showed moderate activity of new bone formation with crossing-over of new bone trabeculae along with the presence of resorption cavities in group II, whereas in the control group, the process of new bone formation was active from both ends and the defect site appeared as a homogenous non-fluoroscent area. Angiograms of the animals in the control group showed uniform angiogenesis in the defect site with establishment of trans-transplant angiogenesis, whereas in group II there was complete trans-transplant shunting of blood vessel communication. Porous HAp ceramic prepared by an aqueous combustion technique promoted bone formation over the defect, confirming their biologic osteoconductive property. PMID:18487940

  7. Pure hydroxyapatite phantoms for the calibration of in vivo X-ray fluorescence systems of bone lead and strontium quantification.

    PubMed

    Da Silva, Eric; Kirkham, Brian; Heyd, Darrick V; Pejović-Milić, Ana

    2013-10-01

    Plaster of Paris [poP, CaSO4·(1)/(2) H2O] is the standard phantom material used for the calibration of in vivo X-ray fluorescence (IVXRF)-based systems of bone metal quantification (i.e bone strontium and lead). Calibration of IVXRF systems of bone metal quantification employs the use of a coherent normalization procedure which requires the application of a coherent correction factor (CCF) to the data, calculated as the ratio of the relativistic form factors of the phantom material and bone mineral. Various issues have been raised as to the suitability of poP for the calibration of IVXRF systems of bone metal quantification which include its chemical purity and its chemical difference from bone mineral (a calcium phosphate). This work describes the preparation of a chemically pure hydroxyapatite phantom material, of known composition and stoichiometry, proposed for the purpose of calibrating IVXRF systems of bone strontium and lead quantification as a replacement for poP. The issue with contamination by the analyte was resolved by preparing pure Ca(OH)2 by hydroxide precipitation, which was found to bring strontium and lead levels to <0.7 and <0.3 μg/g Ca, respectively. HAp phantoms were prepared from known quantities of chemically pure Ca(OH)2, CaHPO4·2H2O prepared from pure Ca(OH)2, the analyte, and a HPO4(2-) containing setting solution. The final crystal structure of the material was found to be similar to that of the bone mineral component of NIST SRM 1486 (bone meal), as determined by powder X-ray diffraction spectrometry. PMID:23980923

  8. Efficacy of nano-hydroxyapatite prepared by an aqueous solution combustion technique in healing bone defects of goat.

    PubMed

    Nandi, Samit Kumar; Kundu, Biswanath; Ghosh, Samir Kumar; De, Dipak Kumar; Basu, Debabrata

    2008-06-01

    The present study was undertaken to evaluate porous hydroxyapatite (HAp), the powder of which was prepared by a novel aqueous solution combustion technique, as a bone substitute in healing bone defects in vivo, as assessed by radiologic and histopathologic methods, oxytetracycline labeling, and angiogenic features in Bengal goat. Bone defects were created in the diaphysis of the radius and either not filled (group I) or filled with a HAp strut (group II). The radiologic study in group II showed the presence of unabsorbed implants which acted as a scaffold for new bone growth across the defect, and the quality of healing of the bone defect was almost indistinguishable from the control group, in which the defect was more or less similar, although the newly formed bony tissue was more organized when HAp was used. Histologic methods showed complete normal ossification with development of Haversian canals and well-defined osteoblasts at the periphery in group II, whereas the control group had moderate fibro-collagenization and an adequate amount of marrow material, fat cells, and blood vessels. An oxytetracycline labeling study showed moderate activity of new bone formation with crossing-over of new bone trabeculae along with the presence of resorption cavities in group II, whereas in the control group, the process of new bone formation was active from both ends and the defect site appeared as a homogenous non-fluoroscent area. Angiograms of the animals in the control group showed uniform angiogenesis in the defect site with establishment of trans-transplant angiogenesis, whereas in group II there was complete trans-transplant shunting of blood vessel communication. Porous HAp ceramic prepared by an aqueous combustion technique promoted bone formation over the defect, confirming their biologic osteoconductive property. PMID:18487940

  9. Enhanced fixation of implants by bone ingrowth to titanium fiber mesh: effect of incorporation of hydroxyapatite powder.

    PubMed

    Tsukeoka, Tadashi; Suzuki, Masahiko; Ohtsuki, Chikara; Tsuneizumi, Yoshikazu; Miyagi, Jin; Sugino, Atsushi; Inoue, Takayuki; Michihiro, Ryouichi; Moriya, Hideshige

    2005-10-01

    Tight fixation between bone and implant materials is of great importance for a successful outcome of procedures such as total knee arthroplasty (TKA) and total hip arthroplasty (THA). Titanium fiber mesh is an attractive structure for the establishment of tight fixation between bone and implant by bone ingrowth into the spaces among the fibers. Enhancement of bone ingrowth is desired not only for tight fixation but also for a fast recovery. Our hypothesis is that just the presence of hydroxyapatite (HA) particles ensures improved bone ingrowth, and that long-term stability can be obtained by mechanical anchoring of bone in the spaces among titanium fibers. In this study, we examine our hypothesis by in vivo experiment using dog femur. HA particles were incorporated in titanium fiber mesh coated on titanium alloy rod by dipping in a slurry of HA with hydroxy-propyl-cellulose in an ethanol solution. Specimens were implanted for 3, 5, and 8 weeks, and were then compared with the results from specimens without the use of HA. Bonding strength was evaluated by push-out test, and histomorphometric measurements were made with analysis software to calculate the average value of bone ingrowth. A significantly higher bonding strength was observed for the specimens with HA-incorporated implant at 3 and 5 weeks, and larger bone ingrowth deep inside the titanium fiber mesh was measured at 3 weeks. Our proposed method has the additional advantage of not requiring a high temperature that may result in changes in characters of HA powder such as phase transition, grain growth, and decomposition. Moreover, this technique of HA powder incorporation without high-temperature treatment allows the use of several types of metallic fiber mesh, as well as the application to fiber mesh made of organic polymers. We conclude that this simple modification of titanium fiber mesh with HA powder can improve the fixation of implant to bone in the initial stage after operation. PMID:16025466

  10. Spatial distribution and survival of human and goat mesenchymal stromal cells on hydroxyapatite and β-tricalcium phosphate.

    PubMed

    Prins, Henk-Jan; Fernandes, Hugo; Rozemuller, Henk; van Blitterswijk, Clemens; de Boer, Jan; Martens, Anton C M

    2016-03-01

    The combination of scaffolds and mesenchymal stromal cells (MSCs) is a promising approach in bone tissue engineering (BTE). Knowledge on the survival, outgrowth and bone-forming capacity of MSCs in vivo is limited. Bioluminescence imaging (BLI), histomorphometry and immunohistochemistry were combined to study the fate of gene-marked goat and human MSCs (gMSCs, hMSCs) on scaffolds with different osteoinductive properties. Luciferase-GFP-labelled MSCs were seeded on hydroxyapatite (HA) or β-tricalcium phosphate (TCP), cultured for 7 days in vitro in osteogenic medium, implanted subcutaneously in immunodeficient mice and monitored with BLI for 6 weeks. The constructs were retrieved and processed for histomorphometry and detection of luciferase-positive cells (LPCs). For gMSCs, BLI revealed doubling of signal after 1 week, declining to 60% of input after 3 weeks and remaining constant until week 6. hMSCs showed a constant decrease of BLI signal to 25% of input, indicating no further expansion. Bone formation of gMSCs was two-fold higher on TCP than HA. hMSCs and gMSCs control samples produced equal amounts of bone on TCP. Upon transduction, there was a four-fold reduction in bone formation compared with untransduced hMSCs, and no bone was formed on HA. LPCs were detected at day 14, but were much less frequent at day 42. Striking differences were observed in spatial distribution. MSCs in TCP were found to be aligned and interconnected on the surface but were scattered in an unstructured fashion in HA. In conclusion, the spatial distribution of MSCs on the scaffold is critical for cell-scaffold-based BTE. PMID:23255230

  11. Three-dimensional poly (ε-caprolactone)/hydroxyapatite/collagen scaffolds incorporating bone marrow mesenchymal stem cells for the repair of bone defects.

    PubMed

    Qi, Xin; Huang, Yinjun; Han, Dan; Zhang, Jieyuan; Cao, Jiaqing; Jin, Xiangyun; Huang, Jinghuan; Li, Xiaolin; Wang, Ting

    2016-04-01

    We previously demonstrated that three-dimensional (3D) hydroxyapatite (HAP)-collagen (COL)-coated poly(ε-caprolactone) (PCL) scaffolds (HAP-COL-PCL) possess appropriate nano-structures, surface roughness, and nutrients, providing a favorable environment for osteogenesis. However, the effect of using 3D HAP-COL-PCL scaffolds incorporating BMSCs for the repair of bone defects in rats has been not evaluated. 3D PCL scaffolds coated with HAP, collagen or HAP/COL and incorporating BMSCs were implanted into calvarial defects. At 12 weeks after surgery, the rats were sacrificed and crania were harvested to assess the bone defect repair using microcomputed tomography (micro-CT), histology, immunohistochemistry and sequential fluorescent labeling analysis. 3D micro-CT reconstructed images and quantitative analysis showed that HAP-COL-PCL groups possessed better bone-forming capacity than HAP-PCL groups or COL-PCL groups. Fluorescent labeling analysis revealed the percentage of tetracycline labeling, alizarin red labeling, and calcein labeling in HAP-COL-PCL groups were all greater than in the other two groups (P < 0.05), and the result was confirmed by immunohistochemical staining and histological analysis of bone regeneration. This study demonstrates that 3D HAP-COL-PCL scaffolds incorporating BMSCs markedly enhance bone regeneration of bone defects in rats. PMID:26964015

  12. Fabrication and in vitro biological evaluation of photopolymerisable hydroxyapatite hydrogel composites for bone regeneration.

    PubMed

    Killion, John A; Geever, Luke M; Devine, Declan M; Higginbotham, Clement L

    2014-04-01

    The aim of this study was to improve the bioactive and compressive properties of photopolymerisable polyethylene glycol hydrogels with the incorporation of hydroxyapatite at different loadings. The synthesis of pure hydroxyapatite was verified through Fourier transform infrared spectroscopy (FTIR) analysis by the complete reaction of all constituents. The formation of a bioactive layer of the hydrogel based composites was confirmed through the formation of carbonate hydroxyapatite after soaking the samples in simulated body fluid. The incorporation of hydroxyapatite into the system resulted in an increase in Young's modulus from 4.36 to 12.73 MPa and an increase in the stress at limit value from 1.20 to 4.42 MPa. This was due to the hydroxyapatite absorbing the compressive load, the polymer matrix distributing the load, a reduction in swelling and the presence of physical crosslinking between both components. Drug dissolution testing showed that the release rate of a drug from the hydrogels was dependent on the molecular weight of the polymer and the type of drug used. PMID:24114559

  13. The effect on bone growth enhancement of implant coatings with hydroxyapatite and collagen deposited electrochemically and by plasma spray

    PubMed Central

    Daugaard, Henrik; Elmengaard, Brian; Bechtold, Joan E.; Jensen, Thomas; Soballe, Kjeld

    2013-01-01

    Skeletal bone consists of hydroxyapatite (HA) [Ca10(PO4)6(OH)2] and collagen type I, both of which are osseoconductive. The goal of osseointegration of orthopedic and dental implants is the rapid achievement of a mechanically stable long-lasting fixation between bone and an implant surface. In this study, we evaluated the mechanical fixation and tissue distribution surrounding implants coated with three surfaces: plasma-sprayed HA coating, thinner coating of electrochemical-assisted deposition of HA, and an identical thin coating with a top layer of mineralized collagen. Uncoated plasma-sprayed titanium (Ti-6Al-4V) served as negative control. The electrochemical-assisted deposition was performed near physiological conditions. We used a canine experimental joint replacement model with four cylindrical implants (one of each treatment group) inserted in the humeri cancellous metaphyseal bone in a 1 mm gap. Observation time was 4 weeks. The mechanical fixation was quantified by push-out test to failure, and the peri-implant tissue formation by histomorphometric evaluation. HA coatings deposited by plasma spray technique or electrochemically, increased the mechanical fixation and bone ongrowth, but there was no statistical difference between the individual HA applications. Addition of collagen to the mineralized phase of the coating to create a more bone natural surface did not improve the osseoconductive effect of HA. PMID:19291683

  14. Bone plate composed of a ternary nano-hydroxyapatite/polyamide 66/glass fiber composite: biomechanical properties and biocompatibility.

    PubMed

    Qiao, Bo; Li, Jidong; Zhu, Qingmao; Guo, Shuquan; Qi, Xiaotong; Li, Weichao; Wu, Jun; Liu, Yang; Jiang, Dianming

    2014-01-01

    An ideal bone plate for internal fixation of bone fractures should have good biomechanical properties and biocompatibility. In this study, we prepared a new nondegradable bone plate composed of a ternary nano-hydroxyapatite/polyamide 66/glass fiber (n-HA/PA66/GF) composite. A breakage area on the n-HA/PA66/GF plate surface was characterized by scanning electron microscopy. Its mechanical properties were investigated using bone-plate constructs and biocompatibility was evaluated in vitro using bone marrow-derived mesenchymal stem cells. The results confirmed that adhesion between the n-HA/PA66 matrix and the glass fibers was strong, with only a few fibers pulled out at the site of breakage. Fractures fixed by the n-HA/PA66/GF plate showed lower stiffness and had satisfactory strength compared with rigid fixation using a titanium plate. Moreover, the results with regard to mesenchymal stem cell morphology, MTT assay, Alizarin Red S staining, enzyme-linked immunosorbent assay, and reverse transcription polymerase chain reaction for alkaline phosphatase and osteocalcin showed that the n-HA/PA66/GF composite was suitable for attachment and proliferation of mesenchymal stem cells, and did not have a negative influence on matrix mineralization or osteogenic differentiation of mesenchymal stem cells. These observations indicate that the n-HA/PA66/GF plate has good biomechanical properties and biocompatibility, and may be considered a new option for internal fixation in orthopedic surgery. PMID:24669191

  15. Injectable porous nano-hydroxyapatite/chitosan/tripolyphosphate scaffolds with improved compressive strength for bone regeneration.

    PubMed

    Uswatta, Suren P; Okeke, Israel U; Jayasuriya, Ambalangodage C

    2016-12-01

    In this study we have fabricated porous injectable spherical scaffolds using chitosan biopolymer, sodium tripolyphosphate (TPP) and nano-hydroxyapatite (nHA). TPP was primarily used as an ionic crosslinker to crosslink nHA/chitosan droplets. We hypothesized that incorporating nHA into chitosan could support osteoconduction by emulating the mineralized cortical bone structure, and improve the Ultimate Compressive Strength (UCS) of the scaffolds. We prepared chitosan solutions with 0.5%, 1% and 2% (w/v) nHA concentration and used simple coacervation and lyophilization techniques to obtain spherical scaffolds. Lyophilized spherical scaffolds had a mean diameter of 1.33mm (n=25). Further, portion from each group lyophilized scaffolds were soaked and dried to obtain Lyophilized Soaked and Dried (LSD) scaffolds. LSD scaffolds had a mean diameter of 0.93mm (n=25) which is promising property for the injectability. Scanning Electron Microscopy images showed porous surface morphology and interconnected pore structures inside the scaffolds. Lyophilized and LSD scaffolds had surface pores <10 and 2μm, respectively. 2% nHA/chitosan LSD scaffolds exhibited UCS of 8.59MPa compared to UCS of 2% nHA/chitosan lyophilized scaffolds at 3.93MPa. Standardize UCS values were 79.98MPa and 357MPa for 2% nHA/chitosan lyophilized and LSD particles respectively. One-way ANOVA results showed a significant increase (p<0.001) in UCS of 1% and 2% nHA/chitosan lyophilized scaffolds compared to 0% and 0.5% nHA/chitosan lyophilized scaffolds. Moreover, 2% nHA LSD scaffolds had significantly increased (p<0.005) their mean UCS by 120% compared to 2% nHA lyophilized scaffolds. In a drawback, all scaffolds have lost their mechanical properties by 95% on the 2nd day when fully immersed in phosphate buffered saline. Additionally live and dead cell assay showed no cytotoxicity and excellent osteoblast attachment to both lyophilized and LSD scaffolds at the end of 14th day of in vitro studies. 2% n

  16. Bone integration capability of a series of strontium-containing hydroxyapatite coatings formed by micro-arc oxidation.

    PubMed

    Yan, Jun; Sun, Ji-Feng; Chu, Paul K; Han, Yong; Zhang, Yu-Mei

    2013-09-01

    Strontium-containing hydroxyapatites (Sr-HA) combine the desirable bone regenerative properties of hydroxyapatites (HA) with anabolic and anti-catabolic effects of strontium cations. In the present work, a series of Sr(y)HA [Sr(y)Ca(10-y)(PO4)6(OH)2; y = 0, 0.5, 1, 2] coatings on titanium are produced by micro-arc oxidation (MAO), and the effects of the in vivo osseointegration ability of the coatings are investigated by using a rabbit model. All samples are subjected to biomechanical, surface elemental, micro-CT and histological analysis after 4 and 12 weeks of healing. The obtained results show that the MAO-formed coatings exhibit a microporous network structure composed of Sr(y)HA/Sr(y)HA-Sr(x)Ca(1-x)TiO3/Sr(x)Ca(1-x)TiO3-TiO2 multilayers, in which the outer Sr(y)HA and intermediate Sr(y)HA-Sr(x)Ca(1-x)TiO3 layers have a nanocrystalline structure. All Sr-HA coated implants induce marked improvements in the behavior of bone formation, quantity and quality of bone tissue around the implants than the control HA implant and in particular, the 20%Sr-HA coating promotes early bone formation as identified by polyfluorochrome sequential labeling. The bone-to-implant contact is increased by 46% (p < 0.05) and the pull-out strength is increased by 103% over the HA group (p < 0.01). Extensive areas of mineralized tissue densely deposit on the 20%Sr-HA coating after biomechanical testing, and the greatest improvement of bone microarchitecture are observed around the 20%Sr-HA implant. The identified biological parameters successfully demonstrate the osteoconductivity of 20%Sr-HA surfaces, which results not only in an acceleration but also an improvement of bone-implant integration. The study demonstrates the immense potential of 20%Sr-HA coatings in dental and orthopedic applications. PMID:23348908

  17. A study on the promotion and suppression of demineralization of human dental hard tissues and hydroxyapatite.

    PubMed

    Odajima, T; Onishi, M

    1998-01-01

    Demineralization of human dental enamel and dentine and their analogue compound, hydroxyapatite, was examined by using pH-metry to measure the time-courses of neutralization of acetate, formate, lactate or propionate buffer solution or of acidification of EDTA solution. The extent of neutralization by enamel, dentine and hydroxyapatite was different for each acid but increased in the same order: propionate, acetate, lactate and formate. This order was consistent with that of the K values of these acids. The pH-metry was used to determine the influences of sodium chloride and sucrose on demineralization of enamel, dentine and hydroxyapatite by acetate, formate, lactate and propionate and by EDTA. The demineralization by these bioorganic acids was suppressed by sucrose but promoted by sodium chloride, except that the demineralization of enamel by acetate and propionate was little affected. The demineralization of enamel, dentine and hydroxyapatite by EDTA was little affected by sucrose but promoted by sodium chloride. The promotive effect of sodium chloride on demineralization may be due to the increasing of solubility product by this salt and the suppressive effect of sucrose may be due to the formation of a calcium saccharate formed from the sucrose reacted with calcium on the surface of apatite crystal and/or the reduction of solubility product by the sucrose. In this study, it was also ascertained that the use of pH-metry made it possible to determine easily the demineralization. PMID:11063021

  18. Gentamicin-loaded strontium-containing hydroxyapatite bioactive bone cement--an efficient bioactive antibiotic drug delivery system.

    PubMed

    Liu, W C; Wong, C T; Fong, M K; Cheung, W S; Kao, R Y T; Luk, K D K; Lu, W W

    2010-11-01

    Modified strontium-containing hydroxyapatite (Sr-HA) bone cement was loaded with gentamicin sulfate to generate an efficient bioactive antibiotic drug delivery system for treatment of bone defects. Gentamicin release and its antibacterial property were determined by fluorometric method and inhibition of Staphylococcus aureus (S. aureus) growth. Gentamicin was released from Sr-HA bone cement during the entire period of study and reached around 38% (w/w) cumulatively after 30 days. Antibacterial activity of the gentamicin loaded in the cements is clearly confirmed by the growth inhibition of S. aureus. The results of the amount and duration of gentamicin release suggest a better drug delivery efficiency in Sr-HA bone cement over polymethylmethacrylate bone cement. Bioactivity of the gentamicin-loaded Sr-HA bone cement was confirmed with the formation of apatite layer with 1.836 ± 0.037 μm thick on day 1 and 5.177 ± 1.355 μm thick on day 7 after immersion in simulated body fluid. Compressive strengths of the gentamicin-loaded Sr-HA cement reached 132.60 ± 10.08 MPa, with a slight decrease from the unloaded groups by 4-9%. Bending moduli of Sr-HA cements with and without gentamicin were 1.782 ± 0.072 GPa and 1.681 ± 0.208 GPa, respectively. On the contrary, unloaded Sr-HA cement obtained slightly larger bending strength of 35.48 ± 2.63 MPa comparing with 33.00 ± 1.65 MPa for loaded cement. No statistical difference was found on the bending strengths and modulus of gentamicin-loaded and -unloaded Sr-HA cements. Sr-HA bone cement loaded with gentamicin was proven to be an efficient drug delivery system with uncompromised mechanical properties and bioactivity. PMID:20878924

  19. Study the bonding mechanism of binders on hydroxyapatite surface and mechanical properties for 3DP fabrication bone scaffolds.

    PubMed

    Wei, Qinghua; Wang, Yanen; Li, Xinpei; Yang, Mingming; Chai, Weihong; Wang, Kai; zhang, Yingfeng

    2016-04-01

    In 3DP fabricating artificial bone scaffolds process, the interaction mechanism between binder and bioceramics power determines the microstructure and macro mechanical properties of Hydroxyapatite (HA) bone scaffold. In this study, we applied Molecular Dynamics (MD) methods to investigating the bonding mechanism and essence of binders on the HA crystallographic planes for 3DP fabrication bone scaffolds. The cohesive energy densities of binders and the binding energies, PCFs g(r), mechanical properties of binder/HA interaction models were analyzed through the MD simulation. Additionally, we prepared the HA bone scaffold specimens with different glues by 3DP additive manufacturing, and tested their mechanical properties by the electronic universal testing machine. The simulation results revealed that the relationship of the binding energies between binders and HA surface is consistent with the cohesive energy densities of binders, which is PAM/HA>PVA/HA>PVP/HA. The PCFs g(r) indicated that their interfacial interactions mainly attribute to the ionic bonds and hydrogen bonds which formed between the polar atoms, functional groups in binder polymer and the Ca, -OH in HA. The results of mechanical experiments verified the relationship of Young׳s modulus for three interaction models in simulation, which is PVA/HA>PAM/HA>PVP/HA. But the trend of compressive strength is PAM/HA>PVA/HA>PVP/HA, this is consistent with the binding energies of simulation. Therefore, the Young׳s modulus of bone scaffolds are limited by the Young׳s modulus of binders, and the compressive strength is mainly decided by the viscosity of binder. Finally, the major reasons for differences in mechanical properties between simulation and experiment were found, the space among HA pellets and the incomplete infiltration of glue were the main reasons influencing the mechanical properties of 3DP fabrication HA bone scaffolds. These results provide useful information in choosing binder for 3DP fabrication

  20. Ectopic bone formation cannot occur by hydroxyapatite/β-tricalcium phosphate bioceramics in green fluorescent protein chimeric mice

    NASA Astrophysics Data System (ADS)

    Cheng, Lijia; Duan, Xin; Xiang, Zhou; Shi, Yujun; Lu, Xiaofeng; Ye, Feng; Bu, Hong

    2012-12-01

    Many studies have shown that calcium phosphate ceramics (CP) have osteoconductive and osteoinductive properties; however, the exact mechanism of bone induction has not yet been reported. This study was performed to investigate if destroying immunological function will influence osteogenesis, to explain the mechanism which is unclear. In this study, twenty C57BL/6 mice were divided into two groups (n = 10), in group 1, a hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ceramic was implanted into both the left and right leg muscles of each mouse; in group 2, ten mice experienced lethal irradiation, then were injected bone marrow (BM) cells from green fluorescent protein (GFP) transgenic mice by tail veil, after bone marrow transplantation (BMT), heart, liver, spleen, lung, kidney, and muscle were harvested for biological analysis, after the GFP chimera model was established successfully, the same HA/β-TCP ceramic was implanted into both leg muscles of each mouse immediately after irradiation. 45 and 90 days after implantation, the ceramics of the two groups were harvested to perform with hematoxylin and eosin (HE) and immunohistochemistry (IHC) staining; the results showed that there was no bone formation in group 2, while new bone tissues were detected in group 1. Our findings suggest that the BM cell from GFP transgenic mice is a good biomarker and it could set a good platform for chimera model; it also shows that BM cell is one of cell resources of bone induction, and destruction of immune function will impede osteoinduction by CP. Overall, our results may shed light on clear mechanism study of bone induction in the future.

  1. Development of hydroxyapatite/calcium silicate composites addressed to the design of load-bearing bone scaffolds.

    PubMed

    Sprio, Simone; Tampieri, Anna; Celotti, Giancarlo; Landi, Elena

    2009-04-01

    This work deals with the preparation of bioactive ceramic composites to be employed for the development of load-bearing bone substitutes, made of hydroxyapatite (Ca(10)(PO(4))(6)(OH)(2), HA) and bioactive dicalcium silicate (Ca(2)SiO(4), C(2)S) as a reinforcing phase. The composite materials were prepared by Fast Hot-Pressing (FHP), which allowed the rapid sintering of monolithic ceramics at temperatures up to 1500 degrees C, well above the commonly adopted temperatures for the consolidation of hydroxyapatite (1200-1300 degrees C). The purpose was to achieve the grain coalescence of both HA and the strengthening phase, so that to obtain a homogeneous ceramic material characterized by controlled phase composition and improved mechanical strength; the dwell time was reduced as much as possible to prevent HA decomposition and excessive grain growth. The most remarkable result, in terms of phase composition, was the absence of any secondary phases in the final ceramics other than HA and C(2)S, even after sintering at 1500 degrees C. The flexure strength of the composite materials was found to be much higher than that of HA alone. Further mechanical characterization was also carried out on HA and composites, sintered in different conditions, to evaluate the elastic properties and fracture toughness, and properties close to those of mineral bone were found. These preliminary results confirmed that composites of HA and Ca(2)SiO(4) are promising for the development of bioactive load-bearing ceramic bone substitutes with controlled phase composition. PMID:19627818

  2. Bone responses to zirconia implants with a thin carbonate-containing hydroxyapatite coating using a molecular precursor method.

    PubMed

    Hirota, Masatsugu; Hayakawa, Tohru; Ohkubo, Chikahiro; Sato, Mitsunobu; Hara, Hiroki; Toyama, Takeshi; Tanaka, Yasuhiro

    2014-08-01

    Thin carbonate-containing hydroxyapatite (CA) films coating partially stabilized zirconia (Y-TZP) were prepared (CA-Y-TZP) to establish a metal-free implant system. CA was coated using a molecular precursor method. The CA film was deposited on the surface of Y-TZP using a precursor solution, which was a mixture of a calcium-ethylenediaminetetraacetic acid (EDTA) complex and phosphate compounds. The deposited CA film was characterized by X-ray diffraction, Fourier transform infrared spectroscopy, and energy dispersive X-ray spectroscopy measurements. A focus ion beam system technique revealed that the thickness of the CA film was less than 1.0 µm. Biological evaluations of CA-Y-TZP were performed by immersion experiments in simulated body fluid (SBF) and implantation experiments in the tibiae and femoral condyles of rabbits. In the SBF immersion experiment, apatite deposition progressed more on CA-Y-TZP at the early stage of immersion than on Y-TZP without the CA coating. Animal experiments revealed that bone formation on CA-Y-TZP was similar with than on Y-TZP. Histomorphometrical evaluations showed a significantly higher bone-to-implant contact ratio and bone mass on CA-Y-TZP after implantation into the femoral trabecular bone of rabbits. Therefore, CA-Y-TZP appears to be applicable as a metal-free implant. PMID:24449291

  3. Polyurethane/fluor-hydroxyapatite nanocomposite scaffolds for bone tissue engineering. Part I: morphological, physical, and mechanical characterization.

    PubMed

    Asefnejad, Azadeh; Behnamghader, Aliasghar; Khorasani, Mohammad Taghi; Farsadzadeh, Babak

    2011-01-01

    In this study, new nano-fluor-hydroxyapatite (nFHA)/polyurethane composite scaffolds were fabricated for potential use in bone tissue engineering. Polyester urethane samples were synthesized from polycaprolactone, hexamethylene diisocyanate, and 1,4-butanediol as chain extender. Nano fluor-hydroxyapatite (nFHA) was successfully synthesized by sol-gel method. The solid-liquid phase separation and solvent sublimation methods were used for preparation of the porous composites. Mechanical properties, chemical structure, and morphological characteristics of the samples were investigated by compressive test, Fourier transform infrared, and scanning electron microscopy (SEM) techniques, respectively. The effect of nFHA powder content on porosity and pore morphology was investigated. SEM images demonstrated that the scaffolds were constituted of interconnected and homogeneously distributed pores. The pore size of the scaffolds was in the range 50-250 μm. The result obtained in this research revealed that the porosity and pore average size decreased and compressive modulus increased with nFHA percentage. Considering morphological, physical, and mechanical properties, the scaffold with a higher ratio of nFHA has suitable potential use in tissue regeneration. PMID:21289986

  4. Environmental pH-controlled loading and release of protein on mesoporous hydroxyapatite nanoparticles for bone tissue engineering.

    PubMed

    Zhang, Ning; Gao, Tianlin; Wang, Yu; Wang, Zongliang; Zhang, Peibiao; Liu, Jianguo

    2015-01-01

    To explore the controlled delivery of protein drugs in micro-environment established by osteoblasts or osteoclasts, the loading/release properties of bovine serum albumin (BSA) depending on pH environment were assessed. The adsorption amounts over mesoporous hydroxyapatite (MHA) or hydroxyapatite (HA) decreased as the pH increased, negatively correlating with zeta-potential values. The adsorption behavior over MHA fits well with the Freundlich and Langmuir models at different pHs. The results suggest that the adsorbed amount of protein on MHA or HA depended on the pH of protein solution. MHA adsorbed BSA at basic pH (MHApH 8.4) exhibited a different release kinetics compared with those in acid and neutral environments (MHApH 4.7 and MHApH 7.4), indicating that the release of protein could be regulated by environmental pH at which MHAs adsorb protein. MHApH 8.4 showed a sustained release for 6h before a gradual release when immersing in acidic environment, which is 2h longer than that in neutral environment. This suggests that MHApH 8.4 showed a more sustained release in acidic environment, which can be established by osteoclasts. The variation of adsorption strength between protein and MHA may be responsible for these behaviors. Our findings may be very useful for the development of MHA applications on both bone repair and protein delivery. PMID:25491972

  5. Nanoscale Bonding between Human Bone and Titanium Surfaces: Osseohybridization

    PubMed Central

    Kang, Seok-Man; Chung, Kyu-Rhim; Kim, Seong-Hun; Ahn, Jae-Pyeong

    2015-01-01

    Until now, the chemical bonding between titanium and bone has been examined only through a few mechanical detachment tests. Therefore, in this study, a sandblasted and acid-etched titanium mini-implant was removed from a human patient after 2 months of placement in order to identify the chemical integration mechanism for nanoscale osseointegration of titanium implants. To prepare a transmission electron microscopy (TEM) specimen, the natural state was preserved as much as possible by cryofixation and scanning electron microscope/focused ion beam (SEM-FIB) milling without any chemical treatment. High-resolution TEM (HRTEM), energy dispersive X-ray spectroscopy (EDS), and scanning TEM (STEM)/electron energy loss spectroscopic analysis (EELS) were used to investigate the chemical composition and structure at the interface between the titanium and bone tissue. HRTEM and EDS data showed evidence of crystalline hydroxyapatite and intermixing of bone with the oxide layer of the implant. The STEM/EELS experiment provided particularly interesting results: carbon existed in polysaccharides, calcium and phosphorus existed as tricalcium phosphate (TCP), and titanium existed as oxidized titanium. In addition, the oxygen energy loss near edge structures (ELNESs) showed a possibility of the presence of CaTiO3. These STEM/EELS results can be explained by structures either with or without a chemical reaction layer. The possible existence of the osseohybridization area and the form of the carbon suggest that reconsideration of the standard definition of osseointegration is necessary. PMID:25667930

  6. Bone Repair with Differentiated Osteoblasts from Adipose-derived Stem Cells in Hydroxyapatite/Tricalcium Phosphate In vivo

    PubMed Central

    Hashemibeni, Batool; Dehghani, Leila; Sadeghi, Farzaneh; Esfandiari, Ebrahim; Gorbani, Masood; Akhavan, Ali; Tahani, Soheil T.; Bahramian, Hamid; Goharian, Vahid

    2016-01-01

    Background: Recently, tissue engineering has developed approaches for repair and restoration of damaged skeletal system based on different scaffolds and cells. This study evaluated the ability of differentiated osteoblasts from adipose-derived stem cells (ADSCs) seeded into hydroxyapatite/tricalcium phosphate (HA-TCP) to repair bone. Methods: In this study, ADSCs of 6 canines were seeded in HA-TCP and differentiated into osteoblasts in osteogenic medium in vitro and bone markers evaluated by reverse transcription polymerase chain reaction (RT-PCR). Scanning electron microscopy (SEM) was applied for detection of cells in the pores of scaffold. HA-TCP with differentiated cells as the test group and without cells as the cell-free group were implanted in separate defected sites of canine's tibia. After 8 weeks, specimens were evaluated by histological, immunohistochemical methods, and densitometry test. The data were analyzed using the SPSS 18 version software. Results: The expression of Type I collagen and osteocalcin genes in differentiated cells were indicated by RT-PCR. SEM results revealed the adhesion of cells in scaffold pores. Formation of trabecular bone confirmed by histological sections that revealed the thickness of bone trabecular was more in the test group. Production of osteopontin in extracellular matrix was indicated in both groups. Densitometry method indicated that strength in the test group was similar to cell-free group and natural bone (P > 0.05). Conclusions: This research suggests that ADSCs-derived osteoblasts in HA-TCP could be used for bone tissue engineering and repairing. PMID:27141281

  7. In Vitro characterization of low modulus linoleic acid coated strontium-substituted hydroxyapatite containing PMMA bone cement.

    PubMed

    Lam, W M; Pan, H B; Fong, M K; Cheung, W S; Wong, K L; Li, Z Y; Luk, K D K; Chan, W K; Wong, C T; Yang, C; Lu, W W

    2011-01-01

    Poly (methyl methacrylate) (PMMA) bone cement is widely used in vertebral body augmentation procedures such as vertebroplasty and balloon kyphoplasty. Filling high modulus PMMA increases the modulus of filled verterbra, increasing the risk of fracture in the adjacent vertebra. On the other hand, in porous PMMA bone cements, wear particle generation and deterioration of mechanical performance are the major drawbacks. This study adopts a new approach by utilizing linoleic acid coated strontium substituted hydroxyapatite nanoparticle (Sr-5 HA) and linoleic acid as plasticizer reducing bone cement's modulus with minimal impact on its strength. We determined the compressive strength (UCS) and modulus (Ec), hydrophobicity, injectability, in vitro bioactivity and biocompatibility of this bone cement at different filler and linoleic acid loading. At 20 wt % Sr5-HA incorporation, UCS and Ec were reduced from 63 ± 2 MPa, 2142 ± 129 MPa to 58 ± 2 MPa, 1785 ± 64 MPa, respectively. UCS and Ec were further reduced to 49 ± 2 MPa and 774 ± 70 MPa respectively when 15 v/v of linoleic acid was incorporated. After 7 days of incubation, pre-osteoblast cells (MC3T3-E1) attached on 20 wt % Sr5-HA and 20 wt % Sr5-HA with 15 v/v of linoleic acid group were higher (3.73 ± 0.01 x 10⁴, 2.27 ± 0.02 x 10⁴) than their PMMA counterpart (1.83 ± 0.04 x 10⁴). Incorporation of Sr5-HA with linoleic acid in monomer phase is more effective in reducing the bone cement's stiffness than Sr5-HA alone. Combination of low stiffness and high mechanical strength gives the novel bone cement the potential for use in vertebroplasty cement applications. PMID:21053263

  8. A preclinical study of stem subsidence and graft incorporation after femoral impaction grafting using porous hydroxyapatite as a bone graft extender.

    PubMed

    Howie, Donald W; McGee, Margaret A; Callary, Stuart A; Carbone, Angelo; Stamenkov, Roumen B; Bruce, Warrick J; Findlay, David M

    2011-10-01

    This preclinical in vivo screening study compared bone graft incorporation and stem subsidence in cemented hemiarthroplasty after femoral impaction bone grafting with either morselized allograft bone (n = 5, control group) or a 1:1 mix of allograft and porous hydroxyapatite ceramics (HA) granules (n = 5, HA group). At 14 weeks, there was excellent bone graft incorporation by bone, and the stems were well fixed in both groups. The median subsidence at the cement-bone interface, measured using radiostereometric analysis, was 0.14 and 0.93 mm in the control and HA groups, respectively. The comparable histologic results between groups and good stem fixation in this study support the conduct of a larger scale investigation of the use of porous HA in femoral impaction bone grafting at revision hip arthroplasty. PMID:21802252

  9. Free form fabricated features on CoCr implants with and without hydroxyapatite coating in vivo: a comparative study of bone contact and bone growth induction.

    PubMed

    Grandfield, Kathryn; Palmquist, Anders; Gonçalves, Stéphane; Taylor, Andy; Taylor, Mark; Emanuelsson, Lena; Thomsen, Peter; Engqvist, Håkan

    2011-04-01

    The current study evaluates the in vivo response to free form fabricated cobalt chromium (CoCr) implants with and without hydroxyapatite (HA) plasma sprayed coatings. The free form fabrication method allowed for integration of complicated pyramidal surface structures on the cylindrical implant. Implants were press fit into the tibial metaphysis of nine New Zealand white rabbits. Animals were sacrificed and implants were removed and embedded. Histological analysis, histomorphometry and electron microscopy studies were performed. Focused ion beam was used to prepare thin sections for high-resolution transmission electron microscopy examination. The fabricated features allowed for effective bone in-growth and firm fixation after 6 weeks. Transmission electron microscopy investigations revealed intimate bone-implant integration at the nanometre scale for the HA coated samples. In addition, histomorphometry revealed a significantly higher bone contact on HA coated implants compared to native CoCr implants. It is concluded that free form fabrication in combination with HA coating improves the early fixation in bone under experimental conditions. PMID:21305340

  10. Accumulation of aluminium in lamellar bone after implantation of titanium plates, Ti-6Al-4V screws, hydroxyapatite granules.

    PubMed

    Zaffe, Davide; Bertoldi, Carlo; Consolo, Ugo

    2004-08-01

    Titanium plates, Ti6Al4V screws and surrounding tissues, and biopsies of hydroxyapatite (Osprovit) grafts of maxillary sinus lifting were investigated to evaluate the release and accumulation of ions. Optical microscopy, SEM and X-ray microanalysis were carried out to evaluate the plates and screws removed from patients presenting inflammation and biopsies. Ions release from metallic appliances or leaching from granules towards soft tissues was observed. An accumulation of aluminium but not titanium was found in soft tissues. A peculiar accumulation of aluminium in the dense lamella of newly formed bone was recorded. The results seem to indicate that biological perturbations may be related to aluminium release from the tested biomaterials. The aluminium content of these biomaterials, its diffusion and accumulation are discussed. Further studies on ion release from biomaterials and aluminium fate in skeletal tissues are suggested. PMID:15020159

  11. Cooling rate effects on thermal, structural, and microstructural properties of bio-hydroxyapatite obtained from bovine bone.

    PubMed

    Ramirez-Gutierrez, Cristian F; Palechor-Ocampo, Anderzon F; Londoño-Restrepo, Sandra M; Millán-Malo, Beatriz M; Rodriguez-García, Mario E

    2016-02-01

    This article is focused on the study of cooling rate effects on the thermal, structural, and microstructural properties of hydroxyapatite (HAp) obtained from bovine bone. A three-step process was used to obtain BIO-HAp: hydrothermal, calcinations, and cooling. Calcined samples in a furnace and cooling in air (HAp-CAir), water (HAp-CW), and liquid nitrogen (HAp-CN2), as well as an air cooled sample inside the furnace (HAp-CFAir), were studied. According to this study, the low cooling rate that was achieved for air cooled samples inside the furnace produce single crystal BIO-HAp with better crystalline quality; other samples exhibited polycrystalline structures forming micron and submicron grains. PMID:25952013

  12. Influence of the fluoride ion in the stability of bone hydroxyapatite achieved in patients after the fluo-calcic therapy.

    PubMed

    Mongiorgi, R; Bertocchi, G; Gnudi, S; Maggi, G; Moroni, A; Coppola, G; Riva di Sanseverino, L

    1991-04-01

    It has been proposed that fluoride ion might actively enter into the basic structural framework of the bone, within its main constituent, the hydroxyapatite (HA), by substituting systematically the OH- ion in the crystal chemical formula Ca5[(PO4)3OH]. Accurate compositional studies have become necessary in order to explain an eventual stabilizing effect. Loss in weight (TG) and chemical reaction (DTG) while varying the temperature have been carried out for the first time on specimens under normal conditions and after fluo-calcic treatment, in parallel with accurate chemical compositional determinations by atomic absorption analysis. Our investigation shows that the new structure present after treatment is mechanically stable and proves more resistant to osteolytic processes. PMID:1910745

  13. Engineering anatomically shaped human bone grafts

    PubMed Central

    Grayson, Warren L.; Fröhlich, Mirjam; Yeager, Keith; Bhumiratana, Sarindr; Chan, M. Ete; Cannizzaro, Christopher; Wan, Leo Q.; Liu, X. Sherry; Guo, X. Edward; Vunjak-Novakovic, Gordana

    2009-01-01

    The ability to engineer anatomically correct pieces of viable and functional human bone would have tremendous potential for bone reconstructions after congenital defects, cancer resections, and trauma. We report that clinically sized, anatomically shaped, viable human bone grafts can be engineered by using human mesenchymal stem cells (hMSCs) and a “biomimetic” scaffold-bioreactor system. We selected the temporomandibular joint (TMJ) condylar bone as our tissue model, because of its clinical importance and the challenges associated with its complex shape. Anatomically shaped scaffolds were generated from fully decellularized trabecular bone by using digitized clinical images, seeded with hMSCs, and cultured with interstitial flow of culture medium. A bioreactor with a chamber in the exact shape of a human TMJ was designed for controllable perfusion throughout the engineered construct. By 5 weeks of cultivation, tissue growth was evidenced by the formation of confluent layers of lamellar bone (by scanning electron microscopy), markedly increased volume of mineralized matrix (by quantitative microcomputer tomography), and the formation of osteoids (histologically). Within bone grafts of this size and complexity cells were fully viable at a physiologic density, likely an important factor of graft function. Moreover, the density and architecture of bone matrix correlated with the intensity and pattern of the interstitial flow, as determined in experimental and modeling studies. This approach has potential to overcome a critical hurdle—in vitro cultivation of viable bone grafts of complex geometries—to provide patient-specific bone grafts for craniofacial and orthopedic reconstructions. PMID:19820164

  14. Intermittent administration of human parathyroid hormone (1-34) increases fixation of strontium-doped hydroxyapatite coating titanium implants via electrochemical deposition in ovariectomized rat femur.

    PubMed

    Tao, Zhou-Shan; Zhou, Wan-Shu; Qiang, Zhou; Tu, Kai-kai; Huang, Zheng-Liang; Xu, Hong-Ming; Sun, Tao; Lv, Yang-Xun; Cui, Wei; Yang, Lei

    2016-02-01

    Previous studies have demonstrated the effect of human parathyroid hormone (1-34) (PTH) or strontium-doped hydroxyapatite coating (Sr-HA) on osteoporotic bone implantation. However, reports about effects of PTH plus Sr-HA on bone osseointegration of titanium implants in a state of osteoporosis were limited. This study was designed to investigate the effects of intermittent administration of human parathyroid hormone (1-34) on strontium-doped hydroxyapatite coating (Sr-HA) implant fixation in ovariectomized (OVX) rats. Twelve weeks after bilateral ovariectomy, all animals were randomly divided into four groups including control group, Sr group, PTH group and PTH+Sr group. Forty OVX rats accepted implant insertion in the distal femurs, control group, and PTH group with HA implants and the Sr group and PTH+Sr group with Sr-HA implants. Animals from PTH group and PTH+Sr group then randomly received PTH (60 µg/kg, 3 times a week) until death at 12 weeks. After 12-week healing period, implants from group PTH+Sr revealed improved osseointegration compared with other treatment groups, which is manifested by the exceeding increase of bone area ratio and bone-to-implant contact, the trabecular microarchitecture and the maximal push-out force displayed by tests like histomorphometry, micro-CT, and biomechanics evaluation. These results demonstrated that PTH+ Sr-HA coatings could enhance implant osseointegration in OVX rats, and suggested the feasibility of using this method to improve implant fixation in osteoporotic bone. PMID:26482573

  15. Biodegradable synthetic bone composites

    DOEpatents

    Liu, Gao; Zhao, Dacheng; Saiz, Eduardo; Tomsia, Antoni P.

    2013-01-01

    The invention provides for a biodegradable synthetic bone composition comprising a biodegradable hydrogel polymer scaffold comprising a plurality of hydrolytically unstable linkages, and an inorganic component; such as a biodegradable poly(hydroxyethylmethacrylate)/hydroxyapatite (pHEMA/HA) hydrogel composite possessing mineral content approximately that of human bone.

  16. Ca L2,3-edge XANES and Sr K-edge EXAFS study of hydroxyapatite and fossil bone apatite

    NASA Astrophysics Data System (ADS)

    Zougrou, I. M.; Katsikini, M.; Brzhezinskaya, M.; Pinakidou, F.; Papadopoulou, L.; Tsoukala, E.; Paloura, E. C.

    2016-08-01

    Upon burial, the organic and inorganic components of hard tissues such as bone, teeth, and tusks are subjected to various alterations as a result of interactions with the chemical milieu of soil, groundwater, and presence of microorganisms. In this study, simulation of the Ca L 2,3-edge X-ray absorption near edge structure (XANES) spectrum of hydroxyapatite, using the CTM4XAS code, reveals that the different symmetry of the two nonequivalent Ca(1) and Ca(2) sites in the unit cell gives rise to specific spectral features. Moreover, Ca L 2,3-edge XANES spectroscopy is applied in order to assess variations in fossil bone apatite crystallinity due to heavy bacterial alteration and catastrophic mineral dissolution, compared to well-preserved fossil apatite, fresh bone, and geologic apatite reference samples. Fossilization-induced chemical alterations are investigated by means of Ca L 2,3-edge XANES and scanning electron microscopy (SEM) and are related to histological evaluation using optical microscopy images. Finally, the variations in the bonding environment of Sr and its preference for substitution in the Ca(1) or Ca(2) sites upon increasing the Sr/Ca ratio is assessed by Sr K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy.

  17. Revisiting bone targeting potential of novel hydroxyapatite based surface modified PLGA nanoparticles of risedronate: Pharmacokinetic and biochemical assessment.

    PubMed

    Rawat, Purnima; Ahmad, Iqbal; Thomas, Shindu C; Pandey, Shweta; Vohora, Divya; Gupta, Sarika; Ahmad, Farhan Jalees; Talegaonkar, Sushama

    2016-06-15

    Hydroxyapatite based biodegradable mPEG-PLGA nanoparticles of risedronate (mPEG-PLGA-RIS-HA) were prepared by water miscible dialysis method for synergistic treatment of osteoporosis. The bone targeting potential of prepared nanoparticles was evaluated by performing the cell viability study and protein estimation in pre-osteoblast cell line (MC3T3E1). Biochemical and in-vivo pharmacokinetic studies on osteoporotic rat model treated with different formulations were performed. Under the biochemical study ALP, TRAP, HxP and Calcium levels were determined. Osteoporotic model treated with prepared nanoparticles indicated significant effect on bone. Pharmacokinetic studies revealed 6-fold and 4-fold increase in the relative bioavailability after intravenous and oral administration of nanoparticles respectively as compared to marketed formulation confirming better effective drug transport. Biochemical investigations also showed a significant change in biomarker level which ultimately lead to bone formation/resorption. A stability analysis has also been carried out according to ICH guidelines (Q1AR2) and shelf life was found to be 1year and 4 months for the prepared formulation. Thus the results of present studies indicated that mPEG-PLGA-RIS-HA NPs has a great potential for sustained delivery of RIS for the treatment and prevention of osteoporosis and to minimize the adverse effects of RIS typically induced by its oral administration. PMID:27113864

  18. Three dimensional melt-deposition of polycaprolactone/bio-derived hydroxyapatite composite into scaffold for bone repair.

    PubMed

    Jiang, Wenbo; Shi, Jun; Li, Wei; Sun, Kang

    2013-01-01

    In this study, three dimensional (3D) polycaprolactone/bio-derived hydroxyapatite (PCL/BHA) composite scaffolds were fabricated by using a melt-deposition system (MDS) for the applications in bone repair. PCL/BHA composites with BHA contents of 0, 10, 20, and 40% were successfully processed into 3D scaffolds by using MDS, while it was failed to fabricate PCL/BHA scaffold with BHA content of 60%. The scaffolds produced were demonstrated to possess the same structures as the predefined with highly uniform and completely interconnected pores. The compressive modulus and strength of the PCL/BHA scaffold increased from 27 to 56 MPa and from 1.9 to 4.5 MPa, respectively, as BHA content increased from 0 to 40%. The wettability of PCL/BHA composite scaffold was also improved with the increase of BHA content. Moreover, the PCL/BHA scaffolds fabricated by MDS showed satisfactory biocompatibility and were capable of being integrated with the surrounding host bone. This study shows the feasibility of fabricating 3D PCL/BHA composite scaffolds with favorable pore structures, mechanical properties, wettability and biocompatibility by using MDS and supports further research of developing novel PCL/BHA composite scaffolds with MDS for the applications in bone repair. PMID:23565866

  19. Ca L2,3-edge XANES and Sr K-edge EXAFS study of hydroxyapatite and fossil bone apatite.

    PubMed

    Zougrou, I M; Katsikini, M; Brzhezinskaya, M; Pinakidou, F; Papadopoulou, L; Tsoukala, E; Paloura, E C

    2016-08-01

    Upon burial, the organic and inorganic components of hard tissues such as bone, teeth, and tusks are subjected to various alterations as a result of interactions with the chemical milieu of soil, groundwater, and presence of microorganisms. In this study, simulation of the Ca L 2,3-edge X-ray absorption near edge structure (XANES) spectrum of hydroxyapatite, using the CTM4XAS code, reveals that the different symmetry of the two nonequivalent Ca(1) and Ca(2) sites in the unit cell gives rise to specific spectral features. Moreover, Ca L 2,3-edge XANES spectroscopy is applied in order to assess variations in fossil bone apatite crystallinity due to heavy bacterial alteration and catastrophic mineral dissolution, compared to well-preserved fossil apatite, fresh bone, and geologic apatite reference samples. Fossilization-induced chemical alterations are investigated by means of Ca L 2,3-edge XANES and scanning electron microscopy (SEM) and are related to histological evaluation using optical microscopy images. Finally, the variations in the bonding environment of Sr and its preference for substitution in the Ca(1) or Ca(2) sites upon increasing the Sr/Ca ratio is assessed by Sr K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy. PMID:27379398

  20. In vivo degradation behavior of Ca-deficient hydroxyapatite coated Mg-Zn-Ca alloy for bone implant application.

    PubMed

    Wang, Huanxin; Guan, Shaokang; Wang, Yisheng; Liu, Hongjian; Wang, Haitao; Wang, Liguo; Ren, Chenxing; Zhu, Shijie; Chen, Kuisheng

    2011-11-01

    In present paper, an in vivo study was carried out on uncoated and calcium-deficient hydroxyapatite (Ca-def HA) coated Mg-Zn-Ca alloy to investigate the effect of Ca-def HA coating on the degradation behavior and bone response of magnesium substrate. Magnesium alloy rods were implanted into rabbit femora and evaluated during 24 weeks implantation. The characterization of both implants indicates that in vivo degradation of the Ca-def HA coating and magnesium substrate occurs almost simultaneously, and in vivo valid life of the coating is about 8 weeks, after that the degradation rate of the coated implants increases obviously. The main reasons for the Ca-def HA coating degradation can be attributed to its reaction with body fluid and the substitution of Mg(2+) ions in Ca-def HA. Histopathological examinations show that the Ca-def HA coating has good osteoconductivity and is in favor of the formation of more new bone on the surface of magnesium alloy. So the Ca-def HA coating could not only slow down in vivo degradation of magnesium alloy but also improve its bone response. PMID:21783346

  1. Hydroxyapatite-coated magnesium implants with improved in vitro and in vivo biocorrosion, biocompatibility, and bone response.

    PubMed

    Kim, Sae-Mi; Jo, Ji-Hoon; Lee, Sung-Mi; Kang, Min-Ho; Kim, Hyoun-Ee; Estrin, Yuri; Lee, Jong-Ho; Lee, Jung-Woo; Koh, Young-Hag

    2014-02-01

    Magnesium and its alloys are candidate materials for biodegradable implants; however, excessively rapid corrosion behavior restricts their practical uses in biological systems. For such applications, surface modification is essential, and the use of anticorrosion coatings is considered as a promising avenue. In this study, we coated Mg with hydroxyapatite (HA) in an aqueous solution containing calcium and phosphate sources to improve its in vitro and in vivo biocorrosion resistance, biocompatibility and bone response. A layer of needle-shaped HA crystals was created uniformly on the Mg substrate even when the Mg sample had a complex shape of a screw. In addition, a dense HA-stratum between this layer and the Mg substrate was formed. This HA-coating layer remarkably reduced the corrosion rate of the Mg tested in a simulated body fluid. Moreover, the biological response, including cell attachment, proliferation and differentiation, of the HA-coated samples was enhanced considerably compared to samples without a coating layer. The preliminary in vivo experiments also showed that the biocorrosion of the Mg implant was significantly retarded by HA coating, which resulted in good mechanical stability. In addition, in the case of the HA-coated implants, biodegradation was mitigated, particularly over the first 6 weeks of implantation. This considerably promoted bone growth at the interface between the implant and bone. These results confirmed that HA-coated Mg is a promising material for biomedical implant applications. PMID:23533169

  2. Near-Infrared (NIR) Spectroscopy of Synthetic Hydroxyapatites and Human Dental Tissues.

    PubMed

    Kolmas, Joanna; Marek, Dariusz; Kolodziejski, Waclaw

    2015-08-01

    Near-infrared spectroscopy (NIR) was used to analyze synthetic hydroxyapatite calcined at various temperatures, synthetic carbonated hydroxyapatite, and human hard dental tissues (enamel and dentin). The NIR bands of those materials in the combination, first-overtone, and second-overtone spectral regions were assigned and evaluated for structural characterization. They were attributed to adsorbed and structural water, structural hydroxyl (OH) groups and surface P-OH groups. The NIR spectral features were quantitatively discussed in view of proton solid-state magic-angle spinning nuclear magnetic resonance ((1)H MAS NMR) results. We conclude that the NIR spectra of apatites are useful in the structural characterization of synthetic and biogenic apatites. PMID:26163232

  3. Mesenchymal stem cell responses to bone-mimetic electrospun matrices composed of polycaprolactone, collagen I and nanoparticulate hydroxyapatite.

    PubMed

    Phipps, Matthew C; Clem, William C; Catledge, Shane A; Xu, Yuanyuan; Hennessy, Kristin M; Thomas, Vinoy; Jablonsky, Michael J; Chowdhury, Shafiul; Stanishevsky, Andrei V; Vohra, Yogesh K; Bellis, Susan L

    2011-01-01

    The performance of biomaterials designed for bone repair depends, in part, on the ability of the material to support the adhesion and survival of mesenchymal stem cells (MSCs). In this study, a nanofibrous bone-mimicking scaffold was electrospun from a mixture of polycaprolactone (PCL), collagen I, and hydroxyapatite (HA) nanoparticles with a dry weight ratio of 50/30/20 respectively (PCL/col/HA). The cytocompatibility of this tri-component scaffold was compared with three other scaffold formulations: 100% PCL (PCL), 100% collagen I (col), and a bi-component scaffold containing 80% PCL/20% HA (PCL/HA). Scanning electron microscopy, fluorescent live cell imaging, and MTS assays showed that MSCs adhered to the PCL, PCL/HA and PCL/col/HA scaffolds, however more rapid cell spreading and significantly greater cell proliferation was observed for MSCs on the tri-component bone-mimetic scaffolds. In contrast, the col scaffolds did not support cell spreading or survival, possibly due to the low tensile modulus of this material. PCL/col/HA scaffolds adsorbed a substantially greater quantity of the adhesive proteins, fibronectin and vitronectin, than PCL or PCL/HA following in vitro exposure to serum, or placement into rat tibiae, which may have contributed to the favorable cell responses to the tri-component substrates. In addition, cells seeded onto PCL/col/HA scaffolds showed markedly increased levels of phosphorylated FAK, a marker of integrin activation and a signaling molecule known to be important for directing cell survival and osteoblastic differentiation. Collectively these results suggest that electrospun bone-mimetic matrices serve as promising degradable substrates for bone regenerative applications. PMID:21346817

  4. Indigenous hydroxyapatite coated and bioactive glass coated titanium dental implant system – Fabrication and application in humans

    PubMed Central

    Mistry, Surajit; Kundu, Debabrata; Datta, Someswar; Basu, Debabrata; Soundrapandian, Chidambaram

    2011-01-01

    Background: The use of different bioactive materials as coating on dental implant to restore tooth function is a growing trend in modern Dentistry. In the present study, hydroxyapatite and the bioactive glass-coated implants were evaluated for their behavior in osseous tissue following implantation in 14 patients. Materials and Methods: Bioactive glass and hydroxyapatite formulated and prepared for coating on Ti-6Al-4V alloy. Hydroxyapatite coating was applied on the implant surface by air plasma spray technique and bioactive glass coating was applied by vitreous enameling technique. Their outcome was assessed after 6 months in vivo study in human. Results: Hydroxyapatite and bioactive glass coating materials were nontoxic and biocompatible. Uneventful healing was observed with both types of implants. Conclusion: The results showed bioactive glass is a good alternative coating material for dental implant. PMID:22028507

  5. COMPOSITE POLYMER-COATED MINERAL SCAFFOLDS FOR BONE REGENERATION: FROM MATERIAL CHARACTERIZATION TO HUMAN STUDIES.

    PubMed

    Pertici, G; Carinci, F; Carusi, G; Epistatus, D; Villa, T; Crivelli, F; Rossi, F; Perale, G

    2015-01-01

    Bovine bone xenografts, made of hydroxyapatite (HA), were coated with poly(L-lactide-co-ε- caprolactone) (PLCL) and RGD-containing collagen fragments in order to increase mechanical properties, hydrophilicity, cell adhesion and osteogenicity. In vitro the scaffold microstructure was investigated with Environmental Scanning Electronic Microscopy (ESEM) analysis and micro tomography, while mechanical properties were investigated by means compression tests. In addition, cell attachment and growth within the three-dimensional scaffold inner structure were validated using human osteosarcoma cell lines (SAOS-2 and MG-63). Standard ISO in vivo biocompatibility studies were carried out on model animals, while bone regenerations in humans were performed to assess the efficacy of the product. All results from in vitro to in vivo investigations are here reported, underlining that this scaffold promotes bone regeneration and has good clinical outcome. PMID:26511194

  6. A Signal-Inducing Bone Cement for Magnetic Resonance Imaging-Guided Spinal Surgery Based on Hydroxyapatite and Polymethylmethacrylate

    SciTech Connect

    Wichlas, Florian Seebauer, Christian J.; Schilling, Rene; Rump, Jens; Chopra, Sascha S.; Walter, Thula; Teichgraeber, Ulf K. M.; Bail, Hermann J.

    2012-06-15

    The aim of this study was to develop a signal-inducing bone cement for magnetic resonance imaging (MRI)-guided cementoplasty of the spine. This MRI cement would allow precise and controlled injection of cement into pathologic lesions of the bone. We mixed conventional polymethylmethacrylate bone cement (PMMA; 5 ml methylmethacrylate and 12 g polymethylmethacrylate) with hydroxyapatite (HA) bone substitute (2-4 ml) and a gadolinium-based contrast agent (CA; 0-60 {mu}l). The contrast-to-noise ratio (CNR) of different CA doses was measured in an open 1.0-Tesla scanner for fast T1W Turbo-Spin-Echo (TSE) and T1W TSE pulse sequences to determine the highest signal. We simulated MRI-guided cementoplasty in cadaveric spines. Compressive strength of the cements was tested. The highest CNR was (1) 87.3 (SD 2.9) in fast T1W TSE for cements with 4 {mu}l CA/ml HA (4 ml) and (2) 60.8 (SD 2.4) in T1W TSE for cements with 1 {mu}l CA/ml HA (4 ml). MRI-guided cementoplasty in cadaveric spine was feasible. Compressive strength decreased with increasing amounts of HA from 46.7 MPa (2 ml HA) to 28.0 MPa (4 ml HA). An MRI-compatible cement based on PMMA, HA, and CA is feasible and clearly visible on MRI images. MRI-guided spinal cementoplasty using this cement would permit direct visualization of the cement, the pathologic process, and the anatomical surroundings.

  7. Fabrication and in vivo evaluation of an osteoblast-conditioned nano-hydroxyapatite/gelatin composite scaffold for bone tissue regeneration.

    PubMed

    Samadikuchaksaraei, Ali; Gholipourmalekabadi, Mazaher; Erfani Ezadyar, Elham; Azami, Mahmoud; Mozafari, Masoud; Johari, Behrooz; Kargozar, Saeid; Jameie, Seyed Behnamedin; Korourian, Alireza; Seifalian, Alexander M

    2016-08-01

    In this study, the effects of osteoblast-conditioning on mechanical behavior, biocompatibility, biodegradation and osteoinductive properties of a nano-hydroxyapatite/gelatin (HA/GEL) nanocomposite scaffold was investigated. The scaffold was fabricated using the layer solvent casting combined with the freeze-drying and lamination techniques. The scaffolds were conditioned by culture of osteoblasts on their surface and their elimination by a repeated freeze-thawing process. The potential of the osteoblast-conditioned HA/GEL (HA/GEL/OC) scaffold to support cell adhesion and growth and its cytotoxicity was assessed in vitro using rat mesenchymal stem cells. For in vivo studies, the HA/GEL/OC nanocomposite was implanted in the critical size bone defect created on rat calvarium and studied after 7, 30 and 90 days. The results showed that mechanical and in vitro biological properties of the scaffold were not affected by the process of conditioning. However, in vivo studies demonstrated that osteoblast-conditioning enhanced biocompatibility and osteoinductivity and of the nanocomposite scaffold. The osteoblast conditioning also accelerated collagen content during the bone healing. In the experimental group that received the HA/GEL/OC and MSCs, the newly formed bone occupied almost the entire defect (93.4 ± 3.3%) within 3 months. In conclusion, this study indicates that osteoblast-conditioning is a viable strategy for the development of bone tissue engineering scaffolds. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2001-2010, 2016. PMID:27027855

  8. FACILE SYNTHESIS, CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY OF CELLULOSE-CHITOSAN-HYDROXYAPATITE COMPOSITE MATERIAL, A POTENTIAL MATERIAL FOR BONE TISSUE ENGINEERING

    PubMed Central

    Mututuvari, Tamutsiwa M.; Harkins, April L.

    2013-01-01

    Hydroxyapatite (HAp) is often used as a bone-implant material because it is biocompatible and osteoconductive. However, HAp possesses poor rheological properties and it is inactive against disease-causing microbes. To improve these properties, we developed a green method to synthesize multifunctional composites containing: (1) cellulose (CEL) to impart mechanical strength; (2) chitosan (CS) to induce antibacterial activity thereby maintaining a microbe-free wound site; and (3) HAp. In this method, CS and CEL were co-dissolved in an ionic liquid (IL) and then regenerated from water. HAp was subsequently formed in situ by alternately soaking [CEL+CS] composites in aqueous solutions of CaCl2 and Na2HPO4. At least 88% of IL used was recovered for reuse by distilling the aqueous washings of [CEL+CS]. The composites were characterized using FTIR, XRD and SEM. These composites retained the desirable properties of their constituents. For example, the tensile strength of the composites was enhanced 1.9X by increasing CEL loading from 20% to 80%. Incorporating CS in the composites resulted in composites which inhibited the growth of both Gram positive (MRSA, S. aureus and VRE) and Gram negative (E. coli and P. aeruginosa) bacteria. These findings highlight the potential use of [CEL+CS+HAp] composites as scaffolds in bone tissue engineering. PMID:23595871

  9. Adverse Biological Effect of TiO2 and Hydroxyapatite Nanoparticles Used in Bone Repair and Replacement

    PubMed Central

    Wang, Jiangxue; Wang, Liting; Fan, Yubo

    2016-01-01

    The adverse biological effect of nanoparticles is an unavoidable scientific problem because of their small size and high surface activity. In this review, we focus on nano-hydroxyapatite and TiO2 nanoparticles (NPs) to clarify the potential systemic toxicological effect and cytotoxic response of wear nanoparticles because they are attractive materials for bone implants and are widely investigated to promote the repair and reconstruction of bone. The wear nanoparticles would be prone to binding with proteins to form protein-particle complexes, to interacting with visible components in the blood including erythrocytes, leukocytes, and platelets, and to being phagocytosed by macrophages or fibroblasts to deposit in the local tissue, leading to the formation of fibrous local pseudocapsules. These particles would also be translocated to and disseminated into the main organs such as the lung, liver and spleen via blood circulation. The inflammatory response, oxidative stress, and signaling pathway are elaborated to analyze the potential toxicological mechanism. Inhibition of the oxidative stress response and signaling transduction may be a new therapeutic strategy for wear debris–mediated osteolysis. Developing biomimetic materials with better biocompatibility is our goal for orthopedic implants. PMID:27231896

  10. Low-Temperature Additive Manufacturing of Biomimic Three-Dimensional Hydroxyapatite/Collagen Scaffolds for Bone Regeneration.

    PubMed

    Lin, Kai-Feng; He, Shu; Song, Yue; Wang, Chun-Mei; Gao, Yi; Li, Jun-Qin; Tang, Peng; Wang, Zheng; Bi, Long; Pei, Guo-Xian

    2016-03-23

    Low-temperature additive manufacturing (AM) holds promise for fabrication of three-dimensional (3D) scaffolds containing bioactive molecules and/or drugs. Due to the strict technical limitations of current approaches, few materials are suitable for printing at low temperature. Here, a low-temperature robocasting method was employed to print biomimic 3D scaffolds for bone regeneration using a routine collagen-hydroxyapatite (CHA) composite material, which is too viscous to be printed via normal 3D printing methods at low temperature. The CHA scaffolds had excellent 3D structure and maintained most raw material properties after printing. Compared to nonprinted scaffolds, printed scaffolds promoted bone marrow stromal cell proliferation and improved osteogenic outcome in vitro. In a rabbit femoral condyle defect model, the interconnecting pores within the printed scaffolds facilitated cell penetration and mineralization before the scaffolds degraded and enhanced repair, compared to nonprinted CHA scaffolds. Additionally, the optimal printing parameters for 3D CHA scaffolds were investigated; 600-μm-diameter rods were optimal in terms of moderate mechanical strength and better repair outcome in vivo. This low-temperature robocasting method could enable a variety of bioactive molecules to be incorporated into printed CHA materials and provides a method of bioprinting biomaterials without compromising their natural properties. PMID:26930140

  11. An amphiphilic degradable polymer/hydroxyapatite composite with enhanced handling characteristics promotes osteogenic gene expression in bone marrow stromal cells.

    PubMed

    Kutikov, Artem B; Song, Jie

    2013-09-01

    Electrospun polymer/hydroxyapatite (HA) composites combining biodegradability with osteoconductivity are attractive for skeletal tissue engineering applications. However, most biodegradable polymers such as poly(lactic acid) (PLA) are hydrophobic and do not blend with adequate interfacial adhesion with HA, compromising the structural homogeneity, mechanical integrity and biological performance of the composite. To overcome this challenge, we combined a hydrophilic polyethylene glycol (PEG) block with poly(d,l-lactic acid) to improve the adhesion of the degradable polymer with HA. The amphiphilic triblock copolymer PLA-PEG-PLA (PELA) improved the stability of HA-PELA suspension at 25wt.% HA content, which was readily electrospun into HA-PELA composite scaffolds with uniform fiber dimensions. HA-PELA was highly extensible (failure strain>200% vs. <40% for HA-PLA), superhydrophilic (∼0° water contact angle vs. >100° for HA-PLA), and exhibited an 8-fold storage modulus increase (unlike deterioration for HA-PLA) upon hydration, owing to the favorable interaction between HA and PEG. HA-PELA also better promoted osteochondral lineage commitment of bone marrow stromal cells in unstimulated culture and supported far more potent osteogenic gene expression upon induction than HA-PLA. We demonstrate that the chemical incorporation of PEG is an effective strategy to improve the performance of degradable polymer/HA composites for bone tissue engineering applications. PMID:23791675

  12. Integration of Rabbit Adipose Derived Mesenchymal Stem Cells to Hydroxyapatite Burr Hole Button Device for Bone Interface Regeneration

    PubMed Central

    Gayathri, Viswanathan; Harikrishnan, Varma; Mohanan, Parayanthala Valappil

    2016-01-01

    Adipose Derived Mesenchymal Stem Cells, multipotent stem cells isolated from adipose tissue, present close resemblance to the natural in vivo milieu and microenvironment of bone tissue and hence widely used for in bone tissue engineering applications. The present study evaluates the compatibility of tissue engineered hydroxyapatite burr hole button device (HAP-BHB) seeded with Rabbit Adipose Derived Mesenchymal Stem Cells (ADMSCs). Cytotoxicity, oxidative stress response, apoptotic behavior, attachment, and adherence of adipose MSC seeded on the device were evaluated by scanning electron and confocal microscopy. The results of the MTT (3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide) assay indicated that powdered device material was noncytotoxic up to 0.5 g/mL on cultured cells. It was also observed that oxidative stress related reactive oxygen species production and apoptosis on cell seeded device were similar to those of control (cells alone) except in 3-day period which showed increased reactive oxygen species generation. Further scanning electron and confocal microscopy indicated a uniform attachment of cells and viability up to 200 μm deep inside the device, respectively. Based on the results, it can be concluded that the in-house developed HAP-BHB device seeded with ADMSCs is nontoxic/safe compatible device for biomedical application and an attractive tissue engineered device for calvarial defect regeneration. PMID:26880922

  13. An amphiphilic degradable polymer/hydroxyapatite composite with enhanced handling characteristics promotes osteogenic gene expression in bone marrow stromal cells

    PubMed Central

    Kutikov, Artem B.; Song, Jie

    2013-01-01

    Electrospun polymer/hydroxyapatite (HA) composites combining biodegradability with osteoconductivity are attractive for skeletal tissue engineering applications. However, most biodegradable polymers such as PLA are hydrophobic and do not blend with adequate interfacial adhesion with HA, compromising the structural homogeneity, mechanical integrity, and biological performance of the composite. To overcome this challenge, we incorporated a hydrophilic polyethylene glycol (PEG) block to poly(D,L-lactic acid) to improve the adhesion of the degradable polymer with HA. The amphiphilic triblock copolymer PLA-PEG-PLA (PELA) improved the stability of HA-PELA suspension at 25 wt% HA content, which was readily electrospun into HA-PELA composite scaffolds with uniform fiber dimensions. HA-PELA was highly extensible (failure strain >200% vs. <40% for HA-PLA), superhydrophilic (~0° water contact angle vs. >100° for HA-PLA), and exhibited an 8-fold storage modulus increase (unlike deterioration for HA-PLA) upon hydration, owing to the favorable interaction between HA and PEG. HA-PELA also better promoted osteochondral lineage commitment of bone marrow stromal cells in unstimulated culture and supported far more potent osteogenesis upon induction than HA-PLA. We demonstrate that the chemical incorporation of PEG is an effective strategy to improve the performance of degradable polymer/HA composites for bone tissue engineering applications. PMID:23791675

  14. Fabrication and characterization of novel nano-biocomposite scaffold of chitosan-gelatin-alginate-hydroxyapatite for bone tissue engineering.

    PubMed

    Sharma, Chhavi; Dinda, Amit Kumar; Potdar, Pravin D; Chou, Chia-Fu; Mishra, Narayan Chandra

    2016-07-01

    A novel nano-biocomposite scaffold was fabricated in bead form by applying simple foaming method, using a combination of natural polymers-chitosan, gelatin, alginate and a bioceramic-nano-hydroxyapatite (nHAp). This approach of combining nHAp with natural polymers to fabricate the composite scaffold, can provide good mechanical strength and biological property mimicking natural bone. Environmental scanning electron microscopy (ESEM) images of the nano-biocomposite scaffold revealed the presence of interconnected pores, mostly spread over the whole surface of the scaffold. The nHAp particulates have covered the surface of the composite matrix and made the surface of the scaffold rougher. The scaffold has a porosity of 82% with a mean pore size of 112±19.0μm. Swelling and degradation studies of the scaffold showed that the scaffold possesses excellent properties of hydrophilicity and biodegradability. Short term mechanical testing of the scaffold does not reveal any rupturing after agitation under physiological conditions, which is an indicative of good mechanical stability of the scaffold. In vitro cell culture studies by seeding osteoblast cells over the composite scaffold showed good cell viability, proliferation rate, adhesion and maintenance of osteoblastic phenotype as indicated by MTT assay, ESEM of cell-scaffold construct, histological staining and gene expression studies, respectively. Thus, it could be stated that the nano-biocomposite scaffold of chitosan-gelatin-alginate-nHAp has the paramount importance for applications in bone tissue-engineering in future regenerative therapies. PMID:27127072

  15. Fabrication of novel Si-doped hydroxyapatite/gelatine scaffolds by rapid prototyping for drug delivery and bone regeneration.

    PubMed

    Martínez-Vázquez, F J; Cabañas, M V; Paris, J L; Lozano, D; Vallet-Regí, M

    2015-03-01

    Porous 3-D scaffolds consisting of gelatine and Si-doped hydroxyapatite were fabricated at room temperature by rapid prototyping. Microscopic characterization revealed a highly homogeneous structure, showing the pre-designed porosity (macroporosity) and a lesser in-rod porosity (microporosity). The mechanical properties of such scaffolds are close to those of trabecular bone of the same density. The biological behavior of these hybrid scaffolds is greater than that of pure ceramic scaffolds without gelatine, increasing pre-osteoblastic MC3T3-E1 cell differentiation (matrix mineralization and gene expression). Since the fabrication process of these structures was carried out at mild conditions, an antibiotic (vancomycin) was incorporated in the slurry before the extrusion of the structures. The release profile of this antibiotic was measured in phosphate-buffered saline solution by high-performance liquid chromatography and was adjusted to a first-order release kinetics. Vancomycin released from the material was also shown to inhibit bacterial growth in vitro. The implications of these results for bone tissue engineering applications are discussed. PMID:25560614

  16. A mild one-pot process for synthesising hydroxyapatite/biomolecule bone scaffolds for sustained and controlled antibiotic release.

    PubMed

    Hess, Ulrike; Hill, Sebastian; Treccani, Laura; Streckbein, Philipp; Heiss, Christian; Rezwan, Kurosch

    2015-02-01

    The release of active molecules or the control of nosocomial infections for improved osteoinduction is ideally addressed by a bone substitute material. For this purpose, the feasibility of a mild one-pot process is probed for incorporating directly active proteins and antibiotics in a hydroxyapatite (HAp) based scaffold. The effect of two serum model proteins, bovine serum albumin (BSA) and fibrinogen (FIB), on the microstructure, on selected mechanical properties as well as on degradation behaviour and on protein release are investigated. By protein incorporation, the porosity can be adjusted between 54 and 70% especially due to the foaming ability of BSA. The addition of 5 wt% FIB doubles the biaxial flexural strength up to 6 MPa in comparison to samples without proteins (3 MPa). Protein release experiments show that a rapid release takes place within the first days (between around 3% for FIB and 38% for BSA). As a possible application for osteomyelitis treatment, vancomycin and gentamicin were subsequently added instead of proteins to study their release behaviour and their antibacterial activity, respectively. A controlled antibiotic release was observed for a period of 18 d. By varying the protein type, mixture and quantity, the mechanical strength porosity as well as the protein release and calcium solubility can be controlled. Our studies underpin the suitability of this mild one-pot process as a promising simple-to-use platform for controlled local drug release and bone treatment. PMID:25594361

  17. Surfactant free rapid synthesis of hydroxyapatite nanorods by a microwave irradiation method for the treatment of bone infection

    NASA Astrophysics Data System (ADS)

    Vani, R.; Bharathi Raja, Subramaniya; Sridevi, T. S.; Savithri, K.; Niranjali Devaraj, S.; Girija, E. K.; Thamizhavel, A.; Narayana Kalkura, S.

    2011-07-01

    Mesoporous nanocrystalline hydroxyapatite (nHAp) rods of size 40-75 nm long and 25 nm wide (resembling bone mineral) were synthesized under microwave irradiation without using any surfactants or modifiers. The surface area and average pore size of the nHAp were found to be 32 m2 g - 1 and 4 nm, respectively. Rifampicin (RIF) and ciprofloxacin (CPF) loaded nHAp displayed an initial burst followed by controlled release (zero order kinetics). Combination of CPF and RIF loaded nHAp showed enhanced bacterial growth inhibition against Staphylococcus aureus (S aureus), Staphylococcus epidermidis (S epidermidis) and Escherichia coli (E coli) compared to individual agent loaded nHAp and pure nHAp. In addition, decreased bacterial adhesion (90%) was observed on the surface of CPF plus RIF loaded nHAp. The biocompatibility test toward MG63 cells infected with micro-organisms showed better cell viability and alkaline phosphatase activity (ALP) for the combination of CPF and RIF loaded nHAp. The influence on cell viability of infected MG63 cells was attributed to the simultaneous and controlled release of CPF and RIF from nHAp, which prevented the emergence of subpopulations that were resistant to each other. Hence, apart from the issue of the rapid synthesis of nHAp without surfactants or modifiers, the simultaneous and controlled release of dual drugs from nHAp would be a simple, non-toxic and cost-effective method to treat bone infections.

  18. Molecular interactions in biomineralized hydroxyapatite amino acid modified nanoclay: in silico design of bone biomaterials.

    PubMed

    Katti, Dinesh R; Sharma, Anurag; Ambre, Avinash H; Katti, Kalpana S

    2015-01-01

    A simulations driven approach to design of a novel biomaterial nanocomposite system is described in this study. Nanoclays modified with amino acids (OMMT) were used to mineralize hydroxyapatite (HAP), mimicking biomineralization. Representative models of organically modified montmorillonite clay (OMMT) and OMMT-hydroxyapatite (OMMT-HAP) were constructed using molecular dynamics and validated using X-ray Diffraction (XRD), Fourier Transforms Infrared (FTIR) spectroscopy and Transmission Electron Microscopy (TEM). Attractive interactions exist between Ca atoms of HAP and C=O group of aminovaleric acid, indicating chelate formation in OMMT-HAP. Interaction energy maps describe molecular interactions among different constituents and their quantitative contributions in the OMMT and OMMT-HAP systems at both parallel and perpendicular orientations. High attractive and high repulsive interactions were found between PO4(3-) and MMT clay as well as aminovaleric molecules in OMMT-HAP perpendicular and parallel models. Large non-bonded interactions in OMMT-HAP indicate influence of neighboring environment on PO4(3-) in in situ HAPclay. Extensive hydrogen bonds were observed between functional hydrogen atoms of modifier and MMT clay in OMMT-HAP as compared to OMMT. Thus, HAP interacts with clay through the aminovaleric acid. This computational study provides a framework for materials design and selection for biomaterials used in tissue engineering and other areas of regenerative medicine. PMID:25491979

  19. A human retrieval study of plasma-sprayed hydroxyapatite-coated plateau root form implants after 2 months to 13 years in function.

    PubMed

    Coelho, Paulo G; Bonfante, Estevam A; Marin, Charles; Granato, Rodrigo; Giro, Gabriela; Suzuki, Marcelo

    2010-01-01

    Calcium phosphate-based bioactive ceramics in various physical and chemical formulations have been extensively utilized as biomaterials for bone regeneration/conduction. However, the determination of their in vivo temporal behavior from the short to long term in humans has been a challenge due to the lack of physical reference for morphologic and morphometric evaluation. The present study evaluated bone morphology and morphometry (bone-to-implant contact [BIC]) around plasma-sprayed hydroxyapatite (PSHA)-coated endosseous implants that were retrieved due to prosthetic reasons while successfully in function at the posterior region of the jaws from as early as 2 months to ∼13 years after a 6-month healing period after placement. Bone morphology was evaluated by light microscopy, and BIC was determined using computer software. Irrespective of the time in vivo, lamellar bone was observed in close contact with the implant PSHA-coated surface and between plateaus. BIC ranged from ∼35-95%, was highly directional, and Haversian-like osteonic morphology between plateaus was observed for most implants. The PSHA coating was present with little variation in thickness between the samples retrieved regardless of time in vivo. PMID:21488826

  20. Biocompatible cephalosporin-hydroxyapatite-poly(lactic-co-glycolic acid)-coatings fabricated by MAPLE technique for the prevention of bone implant associated infections

    NASA Astrophysics Data System (ADS)

    Rădulescu, Dragoş; Grumezescu, Valentina; Andronescu, Ecaterina; Holban, Alina Maria; Grumezescu, Alexandru Mihai; Socol, Gabriel; Oprea, Alexandra Elena; Rădulescu, Marius; Surdu, Adrian; Trusca, Roxana; Rădulescu, Radu; Chifiriuc, Mariana Carmen; Stan, Miruna S.; Constanda, Sabrina; Dinischiotu, Anca

    2016-06-01

    In this study we aimed to obtain functionalized thin films based on hydroxyapatite/poly(lactic-co-glycolic acid) (HAp/PLGA) containing ceftriaxone/cefuroxime antibiotics (ATBs) deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The prepared thin films were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-Ray diffraction (XRD), selected area electron diffraction (SAED), and infra red (IR) analysis. HAp/PLGA/ATBs thin films sustained the growth of human osteoblasts, proving their good biocompatibility. The microscopic evaluation and the culture-based quantitative assay of the E. coli biofilm development showed that the thin films inhibited the initial step of microbial attachment as well as the subsequent colonization and biofilm development on the respective surfaces. This study demonstrates that MAPLE technique could represent an appealing technique for the fabrication of antibiotics-containing polymeric implant coatings. The bioevaluation results recommend this type of surfaces for the prevention of bone implant microbial contamination and for the enhanced stimulation of the implant osseointegration process.

  1. Human iPSC-derived osteoblasts and osteoclasts together promote bone regeneration in 3D biomaterials

    PubMed Central

    Jeon, Ok Hee; Panicker, Leelamma M.; Lu, Qiaozhi; Chae, Jeremy J.; Feldman, Ricardo A.; Elisseeff, Jennifer H.

    2016-01-01

    Bone substitutes can be designed to replicate physiological structure and function by creating a microenvironment that supports crosstalk between bone and immune cells found in the native tissue, specifically osteoblasts and osteoclasts. Human induced pluripotent stem cells (hiPSC) represent a powerful tool for bone regeneration because they are a source of patient-specific cells that can differentiate into all specialized cell types residing in bone. We show that osteoblasts and osteoclasts can be differentiated from hiPSC-mesenchymal stem cells and macrophages when co-cultured on hydroxyapatite-coated poly(lactic-co-glycolic acid)/poly(L-lactic acid) (HA–PLGA/PLLA) scaffolds. Both cell types seeded on the PLGA/PLLA especially with 5% w/v HA recapitulated the tissue remodeling process of human bone via coupling signals coordinating osteoblast and osteoclast activity and finely tuned expression of inflammatory molecules, resulting in accelerated in vitro bone formation. Following subcutaneous implantation in rodents, co-cultured hiPSC-MSC/-macrophage on such scaffolds showed mature bone-like tissue formation. These findings suggest the importance of coupling matrix remodeling through osteoblastic matrix deposition and osteoclastic tissue resorption and immunomodulation for tissue development. PMID:27225733

  2. In vitro osteogenic and odontogenic differentiation of human dental pulp stem cells seeded on carboxymethyl cellulose-hydroxyapatite hybrid hydrogel

    PubMed Central

    Teti, Gabriella; Salvatore, Viviana; Focaroli, Stefano; Durante, Sandra; Mazzotti, Antonio; Dicarlo, Manuela; Mattioli-Belmonte, Monica; Orsini, Giovanna

    2015-01-01

    Stem cells from human dental pulp have been considered as an alternative source of adult stem cells in tissue engineering because of their potential to differentiate into multiple cell lineages. Recently, polysaccharide based hydrogels have become especially attractive as matrices for the repair and regeneration of a wide variety of tissues and organs. The incorporation of inorganic minerals as hydroxyapatite nanoparticles can modulate the performance of the scaffolds with potential applications in tissue engineering. The aim of this study was to verify the osteogenic and odontogenic differentiation of dental pulp stem cells (DPSCs) cultured on a carboxymethyl cellulose—hydroxyapatite hybrid hydrogel. Human DPSCs were seeded on carboxymethyl cellulose—hydroxyapatite hybrid hydrogel and on carboxymethyl cellulose hydrogel for 1, 3, 5, 7, 14, and 21 days. Cell viability assay and ultramorphological analysis were carried out to evaluate biocompatibility and cell adhesion. Real Time PCR was carried out to demonstrate the expression of osteogenic and odontogenic markers. Results showed a good adhesion and viability in cells cultured on carboxymethyl cellulose—hydroxyapatite hybrid hydrogel, while a low adhesion and viability was observed in cells cultured on carboxymethyl cellulose hydrogel. Real Time PCR data demonstrated a temporal up-regulation of osteogenic and odontogenic markers in dental pulp stem cells cultured on carboxymethyl cellulose—hydroxyapatite hybrid hydrogel. In conclusion, our in vitro data confirms the ability of DPSCs to differentiate toward osteogenic and odontogenic lineages in presence of a carboxymethyl cellulose—hydroxyapatite hybrid hydrogel. Taken together, our results provide evidence that DPSCs and carboxymethyl cellulose—hydroxyapatite hybrid hydrogel could be considered promising candidates for dental pulp complex and periodontal tissue engineering. PMID:26578970

  3. Human progenitor cells for bone engineering applications.

    PubMed

    de Peppo, G M; Thomsen, P; Karlsson, C; Strehl, R; Lindahl, A; Hyllner, J

    2013-06-01

    In this report, the authors review the human skeleton and the increasing burden of bone deficiencies, the limitations encountered with the current treatments and the opportunities provided by the emerging field of cell-based bone engineering. Special emphasis is placed on different sources of human progenitor cells, as well as their pros and cons in relation to their utilization for the large-scale construction of functional bone-engineered substitutes for clinical applications. It is concluded that, human pluripotent stem cells represent a valuable source for the derivation of progenitor cells, which combine the advantages of both embryonic and adult stem cells, and indeed display high potential for the construction of functional substitutes for bone replacement therapies. PMID:23642054

  4. Coaxial electrospun aligned tussah silk fibroin nanostructured fiber scaffolds embedded with hydroxyapatite-tussah silk fibroin nanoparticles for bone tissue engineering.

    PubMed

    Shao, Weili; He, Jianxin; Sang, Feng; Ding, Bin; Chen, Li; Cui, Shizhong; Li, Kejing; Han, Qiming; Tan, Weilin

    2016-01-01

    The bone is a composite of inorganic and organic materials and possesses a complex hierarchical architecture consisting of mineralized fibrils formed by collagen molecules and coated with oriented hydroxyapatite. To regenerate bone tissue, it is necessary to provide a scaffold that mimics the architecture of the extracellular matrix in native bone. Here, we describe one such scaffold, a nanostructured composite with a core made of a composite of hydroxyapatite and tussah silk fibroin. The core is encased in a shell of tussah silk fibroin. The composite fibers were fabricated by coaxial electrospinning using green water solvent and were characterized using different techniques. In comparison to nanofibers of pure tussah silk, composite notably improved mechanical properties, with 90-fold and 2-fold higher initial modulus and breaking stress, respectively, obtained. Osteoblast-like MG-63 cells were cultivated on the composite to assess its suitability as a scaffold for bone tissue engineering. We found that the fiber scaffold supported cell adhesion and proliferation and functionally promoted alkaline phosphatase and mineral deposition relevant for biomineralization. In addition, the composite were more biocompatible than pure tussah silk fibroin or cover slip. Thus, the nanostructured composite has excellent biomimetic and mechanical properties and is a potential biocompatible scaffold for bone tissue engineering. PMID:26478319

  5. A Prospective Randomized Clinical Trial Comparing Bone Union Rate Following Anterior Cervical Discectomy and Fusion Using a Polyetheretherketone Cage: Hydroxyapatite/B-Tricalcium Phosphate Mixture versus Hydroxyapatite/Demineralized Bone Matrix Mixture

    PubMed Central

    Yi, Jemin; Nam, Woo Dong; Han, Kye Young; Kim, Myung-Ho; Kang, Jong Won; Won, Jonghwa; Kim, Seong Wan; Noh, Won; Yeom, Jin S

    2015-01-01

    Study Design Prospective randomized noninferiority trial. Purpose To evaluate whether the union rate of anterior cervical discectomy and fusion (ACDF) using a polyetheretherketone (PEEK) cage filled with a mixture of hydroxyapatite (HA) and demineralized bone matrix (DBM) is inferior to that of a mixture of β-tricalcium phosphate (β-TCP) and HA. Overview of Literature There have been no clinical trials investigating the outcomes of a mixture of HA and DBM in a PEEK cage in ACDF. Methods Eighty-five eligible patients were randomly assigned to group B (n=43), in which a PEEK cage with a mixture of HA and DBM was used, or group C (n=42), in which a PEEK cage with a mixture of HA and β-TCP was used. The primary study endpoint was the fusion rate, which was assessed with dynamic radiographs and computed tomography (CT) scans. Secondary endpoints included pain intensity using a visual analogue scale, functional outcome using a neck disability index score, laboratory tests of inflammatory profiles, and the infection rate. Results Seventy-seven patients (38 in group B and 39 in group C) were included in the final analysis. One year postoperatively, bone fusion was achieved in 87% of group B patients and 87% of group C patients on dynamic radiographs, and 87% of group B patients and 72% of group C patients on CT scans (p=1.00 and 0.16, respectively). There were also no between-groups differences with respect to the secondary endpoints. Conclusions A HA/DBM mixture inside a PEEK cage can provide noninferior outcomes compared to a HA/TCP mixture in ACDF. PMID:25705332

  6. Micro-/Nano- sized hydroxyapatite directs differentiation of rat bone marrow derived mesenchymal stem cells towards an osteoblast lineage

    NASA Astrophysics Data System (ADS)

    Huang, Yan; Zhou, Gang; Zheng, Lisha; Liu, Haifeng; Niu, Xufeng; Fan, Yubo

    2012-03-01

    Regenerative medicine consisting of cells and materials provides a new way for the repair and regeneration of tissues and organs. Nano-biomaterials are highlighted due to their advantageous features compared with conventional micro-materials. The aim of this study is to investigate the effects of micro-/nano- sized hydroxyapatite (μ/n-HA) on the osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs). μ/n-HA were prepared by a microwave synthesizer and precipitation method, respectively. Different sizes of μ/n-HA were characterized by IR, XRD, SEM, TEM and co-cultured with rBMSCs. It was shown that rBMSCs expressed higher levels of osteoblast-related markers by n-HA than μ-HA stimulation. The size of HA is an important factor for affecting the osteogenic differentiation of rBMSCs. This provides a new avenue for mechanistic studies of stem cell differentiation and a new approach to obtain more committed differentiated cells.

  7. Effect of negatively charged cellulose nanofibers on the dispersion of hydroxyapatite nanoparticles for scaffolds in bone tissue engineering.

    PubMed

    Park, Minsung; Lee, Dajung; Shin, Sungchul; Hyun, Jinho

    2015-06-01

    Nanofibrous 2,2,6,6-tetramethylpiperidine-1-oxyl(TEMPO)-oxidized bacterial cellulose (TOBC) was used as a dispersant of hydroxyapatite (HA) nanoparticles in aqueous solution. The surfaces of TOBC nanofibers were negatively charged after the reaction with the TEMPO/NaBr/NaClO system at pH 10 and room temperature. HA nanoparticles were simply adsorbed on the TOBC nanofibers (HA-TOBC) and dispersed well in DI water. The well-dispersed HA-TOBC colloidal solution formed a hydrogel after the addition of gelatin, followed by crosslinking with glutaraldehyde (HA-TOBC-Gel). The chemical modification of the fiber surfaces and the colloidal stability of the dispersion solution confirmed TOBC as a promising HA dispersant. Both the Young's modulus and maximum tensile stress increased as the amount of gelatin increased due to the increased crosslinking of gelatin. In addition, the well-dispersed HA produced a denser scaffold structure resulting in the increase of the Young's modulus and maximum tensile stress. The well-developed porous structures of the HA-TOBC-Gel composites were incubated with Calvarial osteoblasts. The HA-TOBC-Gel significantly improved cell proliferation as well as cell differentiation confirming the material as a potential candidate for use in bone tissue engineering scaffolds. PMID:25910635

  8. A simple approach for synthesis, characterization and bioactivity of bovine bones to fabricate the polyurethane nanofiber containing hydroxyapatite nanoparticles.

    PubMed

    Sheikh, F A; Kanjwal, M A; Macossay, J; Barakat, N A M; Kim, H Y

    2012-01-01

    In the present study, we had introduced polyurethane (PU) nanofibers that contain hydroxyapatite (HAp) nanoparticles (NPs) as a result of an electrospinning process. A simple method that does not depend on additional foreign chemicals had been employed to synthesize HAp NPs through the calcination of bovine bones. Typically, a colloidal gel consisting of HAp/PU had been electrospun to form nanofibers. In this communication, physiochemical aspects of prepared nanofibers were characterized by FE-SEM, TEM and TEM-EDS, which confirmed that nanofibers were well-oriented and good dispersion of HAp NPs, over the prepared nanofibers. Parameters, affecting the utilization of the prepared nanofibers in various nano-biotechnological fields have been studied; for instance, the bioactivity of the produced nanofiber mats was investigated while incubating in simulated body fluid (SBF). The results from incubation of nanofibers, indicated that incorporation of HAp strongly activates the precipitation of the apatite-like particles, because of the HAp NPs act as seed, that accelerate crystallization of the biological HAp from the utilized SBF. PMID:24416082

  9. Chemical Synthesis, Characterization, and Biocompatibility Study of Hydroxyapatite/Chitosan Phosphate Nanocomposite for Bone Tissue Engineering Applications

    PubMed Central

    Pramanik, Nabakumar; Mishra, Debasish; Banerjee, Indranil; Maiti, Tapas Kumar; Bhargava, Parag; Pramanik, Panchanan

    2009-01-01

    A novel bioanalogue hydroxyapatite (HAp)/chitosan phosphate (CSP) nanocomposite has been synthesized by a solution-based chemical methodology with varying HAp contents from 10 to 60% (w/w). The interfacial bonding interaction between HAp and CSP has been investigated through Fourier transform infrared absorption spectra (FTIR) and x-ray diffraction (XRD). The surface morphology of the composite and the homogeneous dispersion of nanoparticles in the polymer matrix have been investigated through scanning electron microscopy (SEM) and transmission electron microscopy (TEM), respectively. The mechanical properties of the composite are found to be improved significantly with increase in nanoparticle contents. Cytotoxicity test using murine L929 fibroblast confirms that the nanocomposite is cytocompatible. Primary murine osteoblast cell culture study proves that the nanocomposite is osteocompatible and highly in vitro osteogenic. The use of CSP promotes the homogeneous distribution of particles in the polymer matrix through its pendant phosphate groups along with particle-polymer interfacial interactions. The prepared HAp/CSP nanocomposite with uniform microstructure may be used in bone tissue engineering applications. PMID:20130797

  10. Characterization and degradation behavior of AZ31 alloy surface modified by bone-like hydroxyapatite for implant applications

    NASA Astrophysics Data System (ADS)

    Wen, Cuilian; Guan, Shaokang; Peng, Li; Ren, Chenxing; Wang, Xiang; Hu, Zhonghua

    2009-04-01

    Hydroxyapatite (HA) coating on AZ31 alloy substrate was prepared by a cathodic electrodeposition method. The as-deposited specimen was then post-treated with hot alkali solution to improve the corrosion resistance and bioactivity for implant applications. The microstructure and composition of HA coating, as well as its degradation behavior in simulated body fluid (SBF) were investigated. It reveals that the as-deposited coating consists of dicalcium phosphate dehydrate (DCPD, CaHPO 4·2H 2O) and HA. While 10 μm-thick nanowhisker HA coatings doped with Na +, Mg +, HPO 42-and CO 32- can be found after NaOH alkali treatment, which exhibits a very similar composition of natural bone. The post-treated coating was composed of needle-like particles with 1000 nm in length and 35 nm in diameter, having a slenderness ratio of about 28.6. Electrochemical tests shows that the Ecorr of Mg substrate significantly increased from -1.6 to -1.42 V after surface modified by HA coatings. There was obvious mass gain on post-treated specimen immersed in SBF during the first 30 days due to the Ca-P-Mg deposition. The HA-coated AZ31 alloy could slow down the degradation rate and effectively induce the deposition of Ca-P-Mg apatite in SBF, showing a good bioactivity.

  11. Micro-/nano- sized hydroxyapatite directs differentiation of rat bone marrow derived mesenchymal stem cells towards an osteoblast lineage.

    PubMed

    Huang, Yan; Zhou, Gang; Zheng, Lisha; Liu, Haifeng; Niu, Xufeng; Fan, Yubo

    2012-04-01

    Regenerative medicine consisting of cells and materials provides a new way for the repair and regeneration of tissues and organs. Nano-biomaterials are highlighted due to their advantageous features compared with conventional micro-materials. The aim of this study is to investigate the effects of micro-/nano- sized hydroxyapatite (μ/n-HA) on the osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs). μ/n-HA were prepared by a microwave synthesizer and precipitation method, respectively. Different sizes of μ/n-HA were characterized by IR, XRD, SEM, TEM and co-cultured with rBMSCs. It was shown that rBMSCs expressed higher levels of osteoblast-related markers by n-HA than μ-HA stimulation. The size of HA is an important factor for affecting the osteogenic differentiation of rBMSCs. This provides a new avenue for mechanistic studies of stem cell differentiation and a new approach to obtain more committed differentiated cells. PMID:22371072

  12. Chondrogenic Regeneration Using Bone Marrow Clots and a Porous Polycaprolactone-Hydroxyapatite Scaffold by Three-Dimensional Printing

    PubMed Central

    Yao, Qingqiang; Wei, Bo; Liu, Nancy; Li, Chenshuang; Guo, Yang; Shamie, Arya Nick; Chen, James; Tang, Cheng; Jin, Chengzhe; Xu, Yan

    2015-01-01

    Scaffolds play an important role in directing three-dimensional (3D) cartilage regeneration. Our recent study reported the potential advantages of bone marrow clots (MC) in promoting extracellular matrix (ECM) scaffold chondrogenic regeneration. The aim of this study is to build a new scaffold for MC, with improved characteristics in mechanics, shaping, and biodegradability, compared to our previous study. To address this issue, this study prepared a 3D porous polycaprolactone (PCL)-hydroxyapatite (HA) scaffold combined with MC (Group A), while the control group (Group B) utilized a bone marrow stem cell seeded PCL-HA scaffold. The results of in vitro cultures and in vivo implantation demonstrated that although an initial obstruction of nutrient exchange caused by large amounts of fibrin and erythrocytes led to a decrease in the ratio of live cells in Group A, these scaffolds also showed significant improvements in cell adhesion, proliferation, and chondrogenic differentiation with porous recanalization in the later culture, compared to Group B. After 4 weeks of in vivo implantation, Group A scaffolds have a superior performance in DNA content, Sox9 and RunX2 expression, cartilage lacuna-like cell and ECM accumulation, when compared to Group B. Furthermore, Group A scaffold size and mechanics were stable during in vitro and in vivo experiments, unlike the scaffolds in our previous study. Our results suggest that the combination with MC proved to be a highly efficient, reliable, and simple new method that improves the biological performance of 3D PCL-HA scaffold. The MC-PCL-HA scaffold is a candidate for future cartilage regeneration studies. PMID:25530453

  13. Hydroxyapatite-TiO(2)-based nanocomposites synthesized in supercritical CO(2) for bone tissue engineering: physical and mechanical properties.

    PubMed

    Salarian, Mehrnaz; Xu, William Z; Wang, Zhiqiang; Sham, Tsun-Kong; Charpentier, Paul A

    2014-10-01

    Calcium phosphate-based nanocomposites offer a unique solution toward producing scaffolds for orthopedic and dental implants. However, despite attractive bioactivity and biocompatibility, hydroxyapatite (HAp) has been limited in heavy load-bearing applications due to its intrinsically low mechanical strength. In this work, to improve the mechanical properties of HAp, we grew HAp nanoplates from the surface of one-dimensional titania nanorod structures by combining a coprecipitation and sol-gel methodology using supercritical fluid processing with carbon dioxide (scCO2). The effects of metal alkoxide concentration (1.1-1.5 mol/L), reaction temperature (60-80 °C), and pressure (6000-8000 psi) on the morphology, crystallinity, and surface area of the resulting nanostructured composites were examined using scanning electron microscopy (SEM), transmission electron microscopy (TEM), powder X-ray diffraction (XRD), and Brunauer-Emmet-Teller (BET) method. Chemical composition of the products was characterized using Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and X-ray absorption near-edge structure (XANES) analyses. HAp nanoplates and HAp-TiO2 nanocomposites were homogeneously mixed within poly(ε-caprolactone) (PCL) to develop scaffolds with enhanced physical and mechanical properties for bone regeneration. Mechanical behavior analysis demonstrated that the Young's and flexural moduli of the PCL/HAp-TiO2 composites were substantially higher than the PCL/HAp composites. Therefore, this new synthesis methodology in scCO2 holds promise for bone tissue engineering with improved mechanical properties. PMID:25184699

  14. Chondrogenic regeneration using bone marrow clots and a porous polycaprolactone-hydroxyapatite scaffold by three-dimensional printing.

    PubMed

    Yao, Qingqiang; Wei, Bo; Liu, Nancy; Li, Chenshuang; Guo, Yang; Shamie, Arya Nick; Chen, James; Tang, Cheng; Jin, Chengzhe; Xu, Yan; Bian, Xiuwu; Zhang, Xinli; Wang, Liming

    2015-04-01

    Scaffolds play an important role in directing three-dimensional (3D) cartilage regeneration. Our recent study reported the potential advantages of bone marrow clots (MC) in promoting extracellular matrix (ECM) scaffold chondrogenic regeneration. The aim of this study is to build a new scaffold for MC, with improved characteristics in mechanics, shaping, and biodegradability, compared to our previous study. To address this issue, this study prepared a 3D porous polycaprolactone (PCL)-hydroxyapatite (HA) scaffold combined with MC (Group A), while the control group (Group B) utilized a bone marrow stem cell seeded PCL-HA scaffold. The results of in vitro cultures and in vivo implantation demonstrated that although an initial obstruction of nutrient exchange caused by large amounts of fibrin and erythrocytes led to a decrease in the ratio of live cells in Group A, these scaffolds also showed significant improvements in cell adhesion, proliferation, and chondrogenic differentiation with porous recanalization in the later culture, compared to Group B. After 4 weeks of in vivo implantation, Group A scaffolds have a superior performance in DNA content, Sox9 and RunX2 expression, cartilage lacuna-like cell and ECM accumulation, when compared to Group B. Furthermore, Group A scaffold size and mechanics were stable during in vitro and in vivo experiments, unlike the scaffolds in our previous study. Our results suggest that the combination with MC proved to be a highly efficient, reliable, and simple new method that improves the biological performance of 3D PCL-HA scaffold. The MC-PCL-HA scaffold is a candidate for future cartilage regeneration studies. PMID:25530453

  15. Influence of Fe3O4 Nanoparticles in Hydroxyapatite Scaffolds on Proliferation of Primary Human Fibroblast Cells

    NASA Astrophysics Data System (ADS)

    Maleki-Ghaleh, H.; Aghaie, E.; Nadernezhad, A.; Zargarzadeh, M.; Khakzad, A.; Shakeri, M. S.; Beygi Khosrowshahi, Y.; Siadati, M. H.

    2016-05-01

    Modern techniques for expanding stem cells play a substantial role in tissue engineering: the raw material that facilitates regeneration of damaged tissues and treats diseases. The environmental conditions and bioprocessing methods are the primary determinants of the rate of cultured stem cell proliferation. Bioceramic scaffolds made of calcium phosphate are effective substrates for optimal cell proliferation. The present study investigates the effects of two bioceramic scaffolds on proliferating cells in culture media. One scaffold was made of hydroxyapatite and the other was a mixture of hydroxyapatite and ferromagnetic material (Fe3O4 nanoparticles). Disk-shaped (10 mm × 2 mm) samples of the two scaffolds were prepared. Primary human fibroblast proliferation was 1.8- and 2.5-fold faster, respectively, when cultured in the presence of hydroxyapatite or ferrous nanoparticle/hydroxyapatite mixtures. Optical microscopy images revealed that the increased proliferation was due to enhanced cell-cell contact. The presence of magnetic Fe3O4 nanoparticles in the ceramic scaffolds significantly increased cell proliferation compared to hydroxyapatite scaffolds and tissue culture polystyrene.

  16. Influence of Fe3O4 Nanoparticles in Hydroxyapatite Scaffolds on Proliferation of Primary Human Fibroblast Cells

    NASA Astrophysics Data System (ADS)

    Maleki-Ghaleh, H.; Aghaie, E.; Nadernezhad, A.; Zargarzadeh, M.; Khakzad, A.; Shakeri, M. S.; Beygi Khosrowshahi, Y.; Siadati, M. H.

    2016-06-01

    Modern techniques for expanding stem cells play a substantial role in tissue engineering: the raw material that facilitates regeneration of damaged tissues and treats diseases. The environmental conditions and bioprocessing methods are the primary determinants of the rate of cultured stem cell proliferation. Bioceramic scaffolds made of calcium phosphate are effective substrates for optimal cell proliferation. The present study investigates the effects of two bioceramic scaffolds on proliferating cells in culture media. One scaffold was made of hydroxyapatite and the other was a mixture of hydroxyapatite and ferromagnetic material (Fe3O4 nanoparticles). Disk-shaped (10 mm × 2 mm) samples of the two scaffolds were prepared. Primary human fibroblast proliferation was 1.8- and 2.5-fold faster, respectively, when cultured in the presence of hydroxyapatite or ferrous nanoparticle/hydroxyapatite mixtures. Optical microscopy images revealed that the increased proliferation was due to enhanced cell-cell contact. The presence of magnetic Fe3O4 nanoparticles in the ceramic scaffolds significantly increased cell proliferation compared to hydroxyapatite scaffolds and tissue culture polystyrene.

  17. Coralline hydroxyapatite in complex acetabular reconstruction.

    PubMed

    Wasielewski, Ray C; Sheridan, Kate C; Lubbers, Melissa A

    2008-04-01

    This retrospective study examined whether a coralline hydroxyapatite bone graft substitute adequately repaired bone defects during complex acetabular reconstructions. Seventeen patients who underwent acetabular revision using Pro Osteon 500 were assessed to determine whether any cups required re-revision, whether bone had incorporated into the coralline hydroxyapatite grafts, and whether the coralline hydroxyapatite grafts resorbed with time. At latest follow-up, no cups required re-revision, but 1 had failed. Radiographic evidence of bone incorporation was observed in every coralline hydroxyapatite graft. Graft resorption was not observed. PMID:19292282

  18. In vitro and in vivo evaluation of a novel nanosize hydroxyapatite particles/poly(ester-urethane) composite scaffold for bone tissue engineering.

    PubMed

    Laschke, M W; Strohe, A; Menger, M D; Alini, M; Eglin, D

    2010-06-01

    Scaffolds for bone tissue engineering should provide an osteoconductive surface to promote the ingrowth of new bone after implantation into bone defects. This may be achieved by hydroxyapatite loading of distinct scaffold biomaterials. Herein, we analyzed the in vitro and in vivo properties of a novel nanosize hydroxyapatite particles/poly(ester-urethane) (nHA/PU) composite scaffold which was prepared by a salt leaching-phase inverse process. Microtomography, scanning electron microscopy and X-ray spectroscopy analyses demonstrated the capability of the material processing to create a three-dimensional porous PU scaffold with nHA on the surface. Compared to nHA-free PU scaffolds (control), this modified scaffold type induced a significant increase in in vitro adsorption of model proteins. In vivo analysis of the inflammatory and angiogenic host tissue response to implanted nHA/PU scaffolds in the dorsal skinfold chamber model indicated that the incorporation of nHA particles into the scaffold material did not affect biocompatibility and vascularization when compared to control scaffolds. Thus, nHA/PU composite scaffolds represent a promising new type of scaffold for bone tissue engineering, combining the flexible material properties of PU with the advantage of an osteoconductive surface. PMID:20004748

  19. A histological study of non-ceramic hydroxyapatite as a bone graft substitute material in the vertical bone augmentation of the posterior mandible using an interpositional inlay technique: A split mouth evaluation.

    PubMed

    Bechara, Karen; Dottore, Alexandre M; Kawakami, Paulo Y; Gehrke, Sergio A; Coelho, Paulo G; Piattelli, Adriano; Iezzi, Giovanna; Shibli, Jamil Awad

    2015-11-01

    The aim of this study was to compare the influence of graft material (non-ceramic hydroxyapatite versus autologous bone) on bone behaviour and perform a resonance frequency analysis of implants placed in augmented sites to evaluate stability. For this study, 11 patients with bilateral edentulous areas in the mandibular posterior region were selected. Alveolar augmentation osteotomies were bilaterally (split mouth design) performed. In one hemiarch, the space generated by the osteotomy was grafted with an interpositional intra-oral autologous bone graft (control group). In the other hemiarch, the space generated by the osteotomy was grafted with an interpositional non-ceramic hydroxyapatite (ncHA) (test group). The groups were randomized. After 6 months of healing, a bone sample was retrieved from each side for histological evaluation using a trephine drill that was 2-mm in internal diameter. The implant stability quotient (ISQ) was measured by the resonance frequency immediately following implant placement at baseline and after 6 months of follow-up. Good incorporation of the graft was observed in both groups; however, in the test group, a residual-grafted material was observed. Bone density and marrow spaces were similar between groups. Correlations between the ISQ values and the histometric variables were not observed (p>0.05). The results of this trial suggest that both intra-oral autologous bone and ncHA may be elected as interpositional grafting materials to vertically augment posterior atrophic mandibles. PMID:26325427

  20. Human Adipose-Derived Stem Cells on Rapid Prototyped Three-Dimensional Hydroxyapatite/Beta-Tricalcium Phosphate Scaffold.

    PubMed

    Canciani, Elena; Dellavia, Claudia; Ferreira, Lorena Maria; Giannasi, Chiara; Carmagnola, Daniela; Carrassi, Antonio; Brini, Anna Teresa

    2016-05-01

    In the study, we assess a rapid prototyped scaffold composed of 30/70 hydroxyapatite (HA) and beta-tricalcium-phosphate (β-TCP) loaded with human adipose-derived stem cells (hASCs) to determine cell proliferation, differentiation toward osteogenic lineage, adhesion and penetration on/into the scaffold.In this in vitro study, hASCs isolated from fat tissue discarded after plastic surgery were expanded, characterized, and then loaded onto the scaffold. Cells were tested for: viability assay (Alamar Blue at days 3, 7 and Live/Dead at day 32), differentiation index (alkaline phosphatase activity at day 14), scaffold adhesion (standard error of the mean analysis at days 5 and 18), and penetration (ground sections at day 32).All the hASC populations displayed stemness markers and the ability to differentiate toward adipogenic and osteogenic lineages.Cellular vitality increased between 3 and 7 days, and no inhibitory effect by HA/β-TCP was observed. Under osteogenic stimuli, scaffold increased alkaline phosphatase activity of +243% compared with undifferentiated samples. Human adipose-derived stem cells adhered on HA/β-TCP surface through citoplasmatic extensions that occupied the macropores and built networks among them. Human adipose derived stem cells were observed in the core of HA/β-TCP. The current combination of hASCs and HA/β-TCP scaffold provided encouraging results. If authors' data will be confirmed in preclinical models, the present engineering approach could represent an interesting tool in treating large bone defects. PMID:27092915

  1. Cadmium content of human cancellous bone.

    PubMed

    Knuuttila, M; Lappalainen, R; Olkkonen, H; Lammi, S; Alhava, E M

    1982-01-01

    The cadmium content of human cancellous bone was related to age, sex, bone loss, physical properties, and elemental composition. Bone specimens from the anterior iliac crest were collected from 889 cadavers with a normal mineral status, and from 50 cadavers which had bone loss from chronic diseases and immobilization. The element concentrations were analyzed using atomic absorption spectrophotometry. Bone fluoride levels ere determined with the ion specific electrode, the mineral density with the gamma ray attenuation method, and the compressive strength with a strain transducer. The data were analyzed using multiple linear regression analysis. The mean cadmium content of 0.22 +/- 9.16 micrograms/g dry weight (+/- SD) in the samples did not change with age and its content was slightly greater in males than in females. Furthermore, no statistically significant relation was found in cadmium content to bone loss changes or to the calcium content of bone. The cadmium content had a high statistically significant positive correlation with the strontium and nickel content. PMID:7138079

  2. Cadmium content of human cancellous bone

    SciTech Connect

    Knuuttila, M.; Lappalainen, R.; Olkkonen, H.; Lammi, S.; Albava, E.M.

    1982-09-01

    The cadmium content of human cancellous bone was related to age, sex, bone loss, physical properties, and elemental composition. Bone specimens from the anterior iliac crest were collected from 88 cadavers with a normal mineral status, and from 50 cadavers which had bone loss from chronic diseases and immobilization. The element concentrations were analyzed using atomic absorption spectrophotometry. Bone fluoride levels were determined with the ion specific electrode, the mineral density with the gamma ray attenuation method, and the compressive strength with a strain transducer. The data were analyzed using multiple linear regression analysis. The mean cadmium content of 0.22 +/- 0.16 ..mu..g/g dry weight (+/- SD) in the samples did not change with age and its content was slightly greater in males than in females. Furthermore, no statistically significant relationship was found in cadmium content to bone loss changes or to the calcium content of bone. The cadmium content had a high statistically significant positive correlation with the strontium and nickel content.

  3. Future human bone research in space

    NASA Technical Reports Server (NTRS)

    LeBlanc, A.; Shackelford, L.; Schneider, V.

    1998-01-01

    Skylab crewmembers demonstrated negative calcium (Ca) balance reaching about -300 mg/day by flight day 84. Limited bone density (BMD) measurements documented that bone was not lost equally from all parts of the skeleton. Subsequent BMD studies during long duration Russian flights documented the regional extent of bone loss. These studies demonstrated mean losses in the spine, femur neck, trochanter, and pelvis of about 1%-1.6% with large differences between individuals as well as between bone sites in a given individual. Limited available data indicate postflight bone recovery occurred in some individuals, but may require several years for complete restoration. Long duration bedrest studies showed a similar pattern of bone loss and calcium balance (-180 mg/day) as spaceflight. During long duration bedrest, resorption markers were elevated, formation markers were unchanged, 1,25 vitamin D (VitD) and calcium absorption were decreased, and serum ionized Ca was increased. Although this information is a good beginning, additional spaceflight research is needed to assess architectural and subregional bone changes, elucidate mechanisms, and develop efficient as well as effective countermeasures. Space research poses a number of unique problems not encountered in ground-based laboratory research. Therefore, researchers contemplating human spaceflight research need to consider a number of unique problems related to spaceflight in their experimental design.

  4. Fabrication and characterization of a porous multidomain hydroxyapatite scaffold for bone tissue engineering investigations.

    PubMed

    Buckley, Conor Timothy; O'Kelly, Kevin Unai

    2010-05-01

    Tissue-engineering scaffold-based strategies have suffered from limited cell depth viability when cultured in vitro, with viable cells existing within the outer periphery of the fluid-scaffold interface. This is primarily believed to be due to the lack of nutrient delivery into and waste removal from the inner regions of the scaffold construct. This work develops a hydroxyapatite trimodal porous scaffold architecture (i.e., a scaffold providing a discrete domain for cell occupancy and a separate domain for nutrient delivery) through a freeze drying process. Unidirectional channels (500 microm diameter) were incorporated through CNC machining with total combined apparent porosities of 85.1% +/- 0.22%. Effective diffusion coefficients for the bimodal phase (consisting of micro- and meso-pores, without channels) were also determined (7.9 x 10(-10) m(2) s(-1)). Trimodal scaffolds also demonstrated enhanced permeability values (approximately 18-fold increase) compared with bimodal scaffold architectures. In vitro experiments were used to assess initial seeding efficiency and distribution as well as cell viability. The presence of unidirectional channels significantly enhanced initial cell seeding distribution throughout the scaffold depth, while maintaining relatively high seeding efficiencies (67.7% +/- 2.2% for trimodal, 79.1% +/- 2.1% for bimodal scaffolds). Numerical models demonstrated the effectiveness and efficacy of incorporating channels to increase the core oxygen concentration, with the accuracy of these models improved by using experimentally measured cellular oxygen consumption rates and effective diffusion coefficients. The presence of channels had a positive influence in minimizing the concentration gradients compared with bimodal scaffolds for the same cell density distributions. PMID:20166121

  5. Response of Human Osteoblast to n-HA/PEEK—Quantitative Proteomic Study of Bio-effects of Nano-Hydroxyapatite Composite

    NASA Astrophysics Data System (ADS)

    Zhao, Minzhi; Li, Haiyun; Liu, Xiaochen; Wei, Jie; Ji, Jianguo; Yang, Shu; Hu, Zhiyuan; Wei, Shicheng

    2016-03-01

    Nano-sized hydroxyapatite (n-HA) is considered as a bio-active material, which is often mixed into bone implant material, polyetheretherketone (PEEK). To reveal the global protein expression modulations of osteoblast in response to direct contact with the PEEK composite containing high level (40%) nano-sized hydroxyapatite (n-HA/PEEK) and explain its comprehensive bio-effects, quantitative proteomic analysis was conducted on human osteoblast-like cells MG-63 cultured on n-HA/PEEK in comparison with pure PEEK. Results from quantitative proteomic analysis showed that the most enriched categories in the up-regulated proteins were related to calcium ion processes and associated functions while the most enriched categories in the down-regulated proteins were related to RNA process. This enhanced our understanding to the molecular mechanism of the promotion of the cell adhesion and differentiation with the inhibition of the cell proliferation on n-HA/PEEK composite. It also exhibited that although the calcium ion level of incubate environment hadn’t increased, merely the calcium fixed on the surface of material had influence to intracellular calcium related processes, which was also reflect by the higher intracellular Ca2+ concentration of n-HA/PEEK. This study could lead to more comprehensive cognition to the versatile biocompatibility of composite materials. It further proves that proteomics is useful in new bio-effect discovery.

  6. Response of Human Osteoblast to n-HA/PEEK—Quantitative Proteomic Study of Bio-effects of Nano-Hydroxyapatite Composite

    PubMed Central

    Zhao, Minzhi; Li, Haiyun; Liu, Xiaochen; Wei, Jie; Ji, Jianguo; Yang, Shu; Hu, Zhiyuan; Wei, Shicheng

    2016-01-01

    Nano-sized hydroxyapatite (n-HA) is considered as a bio-active material, which is often mixed into bone implant material, polyetheretherketone (PEEK). To reveal the global protein expression modulations of osteoblast in response to direct contact with the PEEK composite containing high level (40%) nano-sized hydroxyapatite (n-HA/PEEK) and explain its comprehensive bio-effects, quantitative proteomic analysis was conducted on human osteoblast-like cells MG-63 cultured on n-HA/PEEK in comparison with pure PEEK. Results from quantitative proteomic analysis showed that the most enriched categories in the up-regulated proteins were related to calcium ion processes and associated functions while the most enriched categories in the down-regulated proteins were related to RNA process. This enhanced our understanding to the molecular mechanism of the promotion of the cell adhesion and differentiation with the inhibition of the cell proliferation on n-HA/PEEK composite. It also exhibited that although the calcium ion level of incubate environment hadn’t increased, merely the calcium fixed on the surface of material had influence to intracellular calcium related processes, which was also reflect by the higher intracellular Ca2+ concentration of n-HA/PEEK. This study could lead to more comprehensive cognition to the versatile biocompatibility of composite materials. It further proves that proteomics is useful in new bio-effect discovery. PMID:26956660

  7. Isolation of osteocytes from human trabecular bone.

    PubMed

    Prideaux, Matthew; Schutz, Christine; Wijenayaka, Asiri R; Findlay, David M; Campbell, David G; Solomon, Lucian B; Atkins, Gerald J

    2016-07-01

    Osteocytes are essential regulators of bone homeostasis. However, they are difficult to study due to their location within the bone mineralised matrix. Although several techniques have been published for the isolation of osteocytes from mouse bone, no such technique has been described for human osteocytes. We have therefore developed a protocol for the isolation of osteocytes from human trabecular bone samples acquired during surgery. The cells were digested from the bone matrix by sequential collagenase and ethylenediaminetetraacetic acid (EDTA) digestions and the cells from later digests displayed characteristic dendritic osteocyte morphology when cultured ex vivo. Furthermore, the cells expressed characteristic osteocyte marker genes, such as E11, dentin matrix protein 1 (DMP1), SOST, matrix extracellular phosphoglycoprotein (MEPE) and phosphate regulating endopeptidase homologue, X-linked (PHEX). In addition, genes associated with osteocyte perilacunar remodelling, including matrix metallopeptidase-13 (MMP13), cathepsin K (CTSK) and carbonic anhydrase 2 (CAR2) were expressed. The cells also responded to parathyroid hormone (PTH) by downregulating SOST mRNA expression and to 1α,25-dihydroxyvitamin D3 (1,25D) by upregulating fibroblast growth factor 23 (FGF23) mRNA expression. Therefore, the cells behave in a similar manner to osteocytes in vivo. These cells represent an important tool in enhancing current knowledge in human osteocyte biology. PMID:27109824

  8. Sphere-shaped nano-hydroxyapatite/chitosan/gelatin 3D porous scaffolds increase proliferation and osteogenic differentiation of human induced pluripotent stem cells from gingival fibroblasts.

    PubMed

    Ji, Jun; Tong, Xin; Huang, Xiaofeng; Wang, Tiancong; Lin, Zitong; Cao, Yazhou; Zhang, Junfeng; Dong, Lei; Qin, Haiyan; Hu, Qingang

    2015-08-01

    Hydroxyapatite (HA) is an important component of human bone and bone tissue engineering scaffolds. A plethora of bone tissue engineering scaffolds have been synthesized so far, including nano-HA/chitosan/gelatin (nHA/CG) scaffolds; and for seeding cells, stem cells, especially induced pluripotent stem cells (iPSCs), have been a promising cell source for bone tissue engineering recently. However, the influence of different HA nano-particle morphologies on the osteogenic differentiation of human iPSCs (hiPSCs) from human gingival fibroblasts (hGFs) is unknown. The purpose of this study was to investigate the osteogenic differentiation of hiPSCs from hGFs seeded on nHA/CG scaffolds with 2 shapes (rod and sphere) of nHA particles. Firstly, hGFs isolated from discarded normal gingival tissues were reprogrammed into hiPSCs. Secondly, hiPSCs were seeded on rod-like nHA/CG (rod-nHA/CG) and sphere-shaped nHA/CG (sphere-nHA/CG) scaffolds respectively and then cell/scaffold complexes were cultured in vitro. Scanning electron microscope, hematoxyline and eosin (HE) staining, Masson's staining, and quantitative real-time polymerase chain reaction techniques were used to examine hiPSC morphology, proliferation, and differentiation on rod-nHA/CG and sphere-nHA/CG scaffolds. Finally, hiPSCs composited with 2 kinds of nHA/CG were transplanted in vivo in a subcutaneous implantation model for 12 weeks; pure scaffolds were also transplanted as a blank control. HE, Masson's, and immunohistochemistry staining were applied to detect new bone regeneration ability. The results showed that sphere-nHA/CG significantly increased hiPSCs from hGF proliferation and osteogenic differentiation in vitro. hiPSCs and sphere-nHA/CG composities generated large bone, whereas hiPSCs and rod-nHA/CG composities produced tiny bone in vivo. Moreover, pure scaffolds without cells almost produced no bone. In conclusion, our work provided a potential innovative bone tissue engineering approach using

  9. Bone formation and degradation behavior of nanocrystalline hydroxyapatite with or without collagen-type 1 in osteoporotic bone defects - an experimental study in osteoporotic goats.

    PubMed

    Alt, Volker; Cheung, Wing Hoi; Chow, Simon K H; Thormann, Ulrich; Cheung, Edmond N M; Lips, Katrin S; Schnettler, Reinhard; Leung, Kwok-Sui

    2016-06-01

    The intention of the current work is to assess new bone formation and degradation behavior of nanocrystalline hydroxyapatite with (HA/col-1) or without collagen-type I (HA) in osteoporotic metaphyseal bone defects in goats. After ovariectomy and special low-calcium diet for three months, 3 drill hole defects in the vertebrae of L3, L4, L5, 4 drill hole defects in the right and left iliac crest and 1 drill hole defect at the distal femur were created in three Chinese mountain goats with a total of 24 defects. The defects were either filled with one of the biomaterials or left empty (empty defect control group). After 42 days, the animals were euthanized and the samples were assessed for new bone formation using high-resolution peripheral quantitative computed tomography (HR-pQCT) and histomorphometry with 2 regions of interest. Detail histology, enzymehistochemistry and immunohistochemistry as well as connexin-43 in situ hybridization and transmission electron microscopy were carried out for evaluation of degradation behavior of the materials and cellular responses of the surrounding tissue in respect to the implants. HR-pQCT showed the highest BV/TV ratio (p = 0.008) and smallest trabecular spacing (p = 0.005) for HA compared to the other groups in the region of interest at the interface with 1mm distance to the initially created defect. The HA/col-1 yielded the highest connectivity density (Conn.D) (p = 0.034) and the highest number of trabeculae (Tb.N) (p = 0.002) compared to the HA and the control group. Histomorphometric analysis for the core region of the initially created defect revealed a statistically higher new bone formation in the HA (p = 0.001) and HA/col-1 group (p = 0.001) compared to the empty defect group including all defect sites. This result was confirmed for site specific analysis with significant higher new bone formation for the HA group for vertebral defects compared to the empty defect group (p = 0.029). For the interface region, no

  10. A process for the development of strontium hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Zahra, N.; Fayyaz, M.; Iqbal, W.; Irfan, M.; Alam, S.

    2014-06-01

    A procedure for the preparation of Strontium Hydroxyapatite is adapted to produce high purity and better homogeneity ceramic with good Crystallinity. The strontium substituted bone cement has potential for use in orthopedic surgeries. Ionic Strontium (Sr) in humans shares the same physiological pathway as calcium and can be deposited in the mineral structure of the bone. In the present study, a novel concept of preparing Sr-contained Hydroxyapatite bone cement by using a precipitation method is proposed to get an ideal biomaterial that possesses potential degradability and more excellent pharmacological effect. Chemical analysis, Fourier Transform Infra Red analysis and Thermogravimetric/ Differential Scanning Calorimetric studies were conducted on prepared Strontium Hydroxyapatite sample to characterize the incorporation of 15% Sr2+ into the crystal lattice of Hydroxyapatite. Strontium was quantitatively incorporated into Hydroxyapatite where its substitution for calcium provoked a linear shift of the infrared absorption bands of the hydroxyl and phosphate groups. Thus, the formation of Sr-HAp was confirmed by Chemical Analysis, FT-IR and TGA/DSC results.

  11. The Osteogenesis of Bone Marrow Stem Cells on mPEG-PCL-mPEG/Hydroxyapatite Composite Scaffold via Solid Freeform Fabrication

    PubMed Central

    Liao, Han-Tsung; Jiang, Cho-Pei

    2014-01-01

    The study described a novel bone tissue scaffold fabricated by computer-aided, air pressure-aided deposition system to control the macro- and microstructure precisely. The porcine bone marrow stem cells (PBMSCs) seeded on either mPEG-PCL-mPEG (PCL) or mPEG-PCL-mPEG/hydroxyapatite (PCL/HA) composite scaffold were cultured under osteogenic medium to test the ability of osteogenesis in vitro. The experimental outcomes indicated that both scaffolds possessed adequate pore size, porosity, and hydrophilicity for the attachment and proliferation of PBMSCs and the PBMSCs expressed upregulated genes of osteogensis and angiogenesis in similar manner on both scaffolds. The major differences between these two types of the scaffolds were the addition of HA leading to higher hardness of PCL/HA scaffold, cell proliferation, and VEGF gene expression in PCL/HA scaffold. However, the in vivo bone forming efficacy between PBMSCs seeded PCL and PCL/HA scaffold was different from the in vitro results. The outcome indicated that the PCL/HA scaffold which had bone-mimetic environment due to the addition of HA resulted in better bone regeneration and mechanical strength than those of PCL scaffold. Therefore, providing a bone-mimetic scaffold is another crucial factor for bone tissue engineering in addition to the biocompatibility, 3D architecture with high porosity, and interpored connection. PMID:24868523

  12. Enhanced healing of rat calvarial defects with sulfated chitosan-coated calcium-deficient hydroxyapatite/bone morphogenetic protein 2 scaffolds.

    PubMed

    Zhao, Jun; Shen, Gang; Liu, Changsheng; Wang, Shaoyi; Zhang, Wenjie; Zhang, Xiaochen; Zhang, Xiuli; Ye, Dongxia; Wei, Jie; Zhang, Zhiyuan; Jiang, Xinquan

    2012-01-01

    Calcium phosphate cements (CPCs), which are widely used in bone regeneration, possess good biocompatibility and osteoconductivity and have been demonstrated to be candidate carriers for bone growth factors. However, limited release of growth factors from CPCs and slow degradation of the materials are not desirable for certain clinical applications. Previous studies have shown that calcium-deficient hydroxyapatite (CDHA) from CPCs presents more rapid degradation rate than CPCs. In this study, a hybrid growth factor delivery system was prepared by using bone morphogenetic protein 2 (BMP-2) loaded CDHA porous scaffold with sulfated chitosan (SCS) coating for improved release profile. We tested the BMP-2 release characteristic of CDHA/BMP-2/SCS composite in vitro and its ability to repair rat calvarial bone defects. A higher percentage of BMP-2 was released when sulfated chitosan coating was present compared with CDHA/BMP-2 group. Eight weeks postoperation, the repaired crania were evaluated by microcomputed tomography, sequential fluorescent labeling, histological analysis, and immunohistochemistry. CDHA/BMP-2/SCS group promoted the most extensive new bone formation than CDHA/BMP-2 and CDHA groups. Our observations suggest that sulfated chitosan coating could enhance the release profile of CDHA/BMP-2 composite in vitro and promote new bone formation in vivo. The hybrid CDHA/BMP-2/SCS system is a promising growth factor delivery strategy for bone regeneration. PMID:21830854

  13. Nanostructured Tendon-Derived Scaffolds for Enhanced Bone Regeneration by Human Adipose-Derived Stem Cells.

    PubMed

    Ko, Eunkyung; Alberti, Kyle; Lee, Jong Seung; Yang, Kisuk; Jin, Yoonhee; Shin, Jisoo; Yang, Hee Seok; Xu, Qiaobing; Cho, Seung-Woo

    2016-09-01

    Decellularized matrix-based scaffolds can induce enhanced tissue regeneration due to their biochemical, biophysical, and mechanical similarity to native tissues. In this study, we report a nanostructured decellularized tendon scaffold with aligned, nanofibrous structures to enhance osteogenic differentiation and in vivo bone formation of human adipose-derived stem cells (hADSCs). Using a bioskiving method, we prepared decellularized tendon scaffolds from tissue slices of bovine Achilles and neck tendons with or without fixation, and investigated the effects on physical and mechanical properties of decellularized tendon scaffolds, based on the types and concentrations of cross-linking agents. In general, we found that decellularized tendon scaffolds without fixative treatments were more effective in inducing osteogenic differentiation and mineralization of hADSCs in vitro. When non-cross-linked decellularized tendon scaffolds were applied together with hydroxyapatite for hADSC transplantation in critical-sized bone defects, they promoted bone-specific collagen deposition and mineralized bone formation 4 and 8 weeks after hADSC transplantation, compared to conventional collagen type I scaffolds. Interestingly, stacking of decellularized tendon scaffolds cultured with osteogenically committed hADSCs and those containing human cord blood-derived endothelial progenitor cells (hEPCs) induced vascularized bone regeneration in the defects 8 weeks after transplantation. Our study suggests that biomimetic nanostructured scaffolds made of decellularized tissue matrices can serve as functional tissue-engineering scaffolds for enhanced osteogenesis of stem cells. PMID:27502160

  14. Improved in vitro biocompatibility of surface-modified hydroxyapatite sponge scaffold with gelatin and BMP-2 in comparison against a commercial bone allograft.

    PubMed

    Carpena, Nathaniel T; Min, Young-Ki; Lee, Byong-Taek

    2015-01-01

    This study aims to demonstrate the morphology and in vitro biocompatibility of neat and surface-modified hydroxyapatite sponge scaffold (SM-HASS) which was fabricated using a sponge replica method, and compared with the commercially available demineralized freeze-dried bone allograft (DFDBA). Surface-modifications were done by coating the surface area of the neat hydroxyapatite sponge scaffold (HASS) with either gelatin alone (HASS/G) or gelatin and BMP-2 growth factor (HASS/G+B). Scanning electron microscope (SEM), Fourier transform infrared (FTIR), porosity, pore size distribution, and compressive strength analyses showed that the addition of gelatin in HASS/G produced a morphologically and structurally similar scaffold to that of the allograft. The addition of BMP-2 improved the biocompatibility of the HASS/G+B in vitro using MC3T3-E1 cells which showed better cell viability, proliferation, and cell adhesion than on the allograft. Therefore, hydroxyapatite scaffold coated with gelatin polymer and gelatin with BMP-2 growth factor showed comparable performance against commercially available DFDBA from cadaver with regards to structure and in vitro biocompatibility. PMID:25248041

  15. Ab initio investigation of the adsorption of zoledronic acid molecule on hydroxyapatite (001) surface: an atomistic insight of bone protection

    NASA Astrophysics Data System (ADS)

    Ri, Mun-Hyok; Yu, Chol-Jun; Jang, Yong-Man; Kim, Song-Un

    2016-03-01

    We report a computational study of the adsorption of zoledronic acid molecule on hydroxyapatite (001) surface within ab initio density functional theory. The systematic study has been performed, from hydroxyapatite bulk and surface, and zoledronic acid molecule to the adsorption of the molecule on the surface. The optimized bond lengths and bond angles were obtained and analyzed, giving an evidence of structural similarity between subjects under study. The formation energies of hydroxyapatite (001) surfaces with two kinds of terminations were computed as about 1.2 and 1.5 J/m^2 with detailed atomistic structural information. We determined the adsorption energies of zoledronic acid molecule on the surfaces, which are -260 kJ/mol at 0.25 ML and -400 kJ/mol at 0.5 ML. An atomistic insight of strong binding affinity of zoledronic acid to the hydroxyapatite surface was given and discussed.

  16. Local delivery of parathyroid hormone-related protein-derived peptides coated onto a hydroxyapatite-based implant enhances bone regeneration in old and diabetic rats.

    PubMed

    Ardura, Juan A; Portal-Núñez, Sergio; Lozano, Daniel; Gutiérrez-Rojas, Irene; Sánchez-Salcedo, Sandra; López-Herradón, Ana; Mulero, Francisca; Villanueva-Peñacarrillo, María L; Vallet-Regí, María; Esbrit, Pedro

    2016-08-01

    Diabetes mellitus (DM) and aging are associated with bone fragility and increased fracture risk. Both (1-37) N- and (107-111) C-terminal parathyroid hormone-related protein (PTHrP) exhibit osteogenic properties. We here aimed to evaluate and compare the efficacy of either PTHrP (1-37) or PTHrP (107-111) loaded into gelatin-glutaraldehyde-coated hydroxyapatite (HA-Gel) foams to improve bone repair of a transcortical tibial defect in aging rats with or without DM, induced by streptozotocin injection at birth. Diabetic old rats showed bone structural deterioration compared to their age-matched controls. Histological and μ-computerized tomography studies showed incomplete bone repair at 4 weeks after implantation of unloaded Ha-Gel foams in the transcortical tibial defects, mainly in old rats with DM. However, enhanced defect healing, as shown by an increase of bone volume/tissue volume and trabecular and cortical thickness and decreased trabecular separation, occurred in the presence of either PTHrP peptide in the implants in old rats with or without DM. This was accompanied by newly formed bone tissue around the osteointegrated HA-Gel implant and increased gene expression of osteocalcin and vascular endothelial growth factor (bone formation and angiogenic markers, respectively), and decreased expression of Sost gene, a negative regulator of bone formation, in the healing bone area. Our findings suggest that local delivery of PTHrP (1-37) or PTHrP (107-111) from a degradable implant is an attractive strategy to improve bone regeneration in aged and diabetic subjects. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2060-2070, 2016. PMID:27086979

  17. Characterization and inhibitive study of gel-grown hydroxyapatite crystals at physiological temperature

    NASA Astrophysics Data System (ADS)

    Parekh, Bharat; Joshi, Mihir; Vaidya, Ashok

    2008-04-01

    Hydroxyapatite is very useful for various biomedical applications, due to its chemical similarity with mineralized bone of human. Hydroxyapatite is also responsible for arthropathy (joint disease). In the present study, the growth of hydroxyapatite crystals was carried out by using single-diffusion gel growth technique in silica hydro gel media, at physiological temperature. The growth of hydroxyapatite crystals under slow and controlled environment in gel medium can be simulated in a simple manner to the growth in human body. The crystals, formed in the Liesegang rings, were characterized by powder XRD, FTIR and dielectric study. The diffusion study is also carried out for the hydroxyapatite crystals using the moving boundary model. The inhibitive influence of various Ayurvedic medicinal plant extracts such as Boswellia serrata gum resin , Tribulus terrestris fruits, Rotula aquatica roots, Boerhaavia diffusa roots and Commiphora wightii, on the growth of hydroxyapatite was studied. Roots of R. aquatica and B. diffusa show some inhibition of the hydroxyapatite crystals in vitro. This preclinical study will be helpful to design the therapy for prevention of hydroxyapatite-based ailments.

  18. Preparation of bone-implants by coating hydroxyapatite nanoparticles on self-formed titanium dioxide thin-layers on titanium metal surfaces.

    PubMed

    Wijesinghe, W P S L; Mantilaka, M M M G P G; Chathuranga Senarathna, K G; Herath, H M T U; Premachandra, T N; Ranasinghe, C S K; Rajapakse, R P V J; Rajapakse, R M G; Edirisinghe, Mohan; Mahalingam, S; Bandara, I M C C D; Singh, Sanjleena

    2016-06-01

    Preparation of hydroxyapatite coated custom-made metallic bone-implants is very important for the replacement of injured bones of the body. Furthermore, these bone-implants are more stable under the corrosive environment of the body and biocompatible than bone-implants made up of pure metals and metal alloys. Herein, we describe a novel, simple and low-cost technique to prepare biocompatible hydroxyapatite coated titanium metal (TiM) implants through growth of self-formed TiO2 thin-layer (SFTL) on TiM via a heat treatment process. SFTL acts as a surface binder of HA nanoparticles in order to produce HA coated implants. Colloidal HA nanorods prepared by a novel surfactant-assisted synthesis method, have been coated on SFTL via atomized spray pyrolysis (ASP) technique. The corrosion behavior of the bare and surface-modified TiM (SMTiM) in a simulated body fluid (SBF) medium is also studied. The highest corrosion rate is found to be for the bare TiM plate, but the corrosion rate has been reduced with the heat-treatment of TiM due to the formation of SFTL. The lowest corrosion rate is recorded for the implant prepared by heat treatment of TiM at 700 °C. The HA-coating further assists in the passivation of the TiM in the SBF medium. Both SMTiM and HA coated SMTiM are noncytotoxic against osteoblast-like (HOS) cells and are in high-bioactivity. The overall production process of bone-implant described in this paper is in high economic value. PMID:27040209

  19. Bone regeneration of hydroxyapatite/alumina bilayered scaffold with 3 mm passage-like medullary canal in canine tibia model.

    PubMed

    Kim, Jong Min; Son, Jun Sik; Kang, Seong Soo; Kim, Gonhyung; Choi, Seok Hwa

    2015-01-01

    The aim of this study was to evaluate the bone regeneration of hydroxyapatite (HA)/alumina bilayered scaffold with a 3 mm passage-like medullary canal in a beagle tibia model. A porous HA/alumina scaffold was fabricated using a polymeric template-coating technique. HA/alumina scaffold dimensions were 10 mm in outer diameter, 20 mm in length, and with either a 3 mm passage or no passage. A 20 mm segmental defect was induced using an oscillating saw through the diaphysis of the beagle tibia. The defects of six beagles were filled with HA/alumina bilayered scaffolds with a 3 mm passage or without. The segmental defect was fixated using one bone plate and six screws. Bone regeneration within the HA/alumina scaffolds was observed at eight weeks after implantation. The evaluation of bone regeneration within the scaffolds after implantation in a beagle tibia was performed using radiography, computerized tomography (CT), micro-CT, and fluorescence microscopy. New bone successfully formed in the tibia defects treated with 3 mm passage HA/alumina scaffolds compared to without-passage HA/alumina scaffolds. It was concluded that the HA/alumina bilayered scaffold with 3 mm passage-like medullary canal was instrumental in inducing host-scaffold engraftment of the defect as well as distributing the newly formed bone throughout the scaffold at 8 weeks after implantation. PMID:25688353

  20. Excavating the Role of Aloe Vera Wrapped Mesoporous Hydroxyapatite Frame Ornamentation in Newly Architectured Polyurethane Scaffolds for Osteogenesis and Guided Bone Regeneration with Microbial Protection.

    PubMed

    Selvakumar, M; Pawar, Harpreet Singh; Francis, Nimmy K; Das, Bodhisatwa; Dhara, Santanu; Chattopadhyay, Santanu

    2016-03-01

    Guided bone regeneration (GBR) scaffolds are unsuccessful in many clinical applications due to a high incidence of postoperative infection. The objective of this work is to fabricate GBR with an anti-infective electrospun scaffold by ornamenting segmented polyurethane (SPU) with two-dimensional Aloe vera wrapped mesoporous hydroxyapatite (Al-mHA) nanorods. The antimicrobial characteristic of the scaffold has been retrieved from the prepared Al-mHA frame with high aspect ratio (∼14.2) via biosynthesis route using Aloe vera (Aloe barbadensis miller) extract. The Al-mHA frame was introduced into an unprecedented SPU matrix (solution polymerized) based on combinatorial soft segments of poly(ε-caprolactone) (PCL), poly(ethylene carbonate) (PEC), and poly(dimethylsiloxane) (PDMS), by an in situ technique followed by electrospinning to fabricate scaffolds. For comparison, pristine mHA nanorods are also ornamented into it. An enzymatic ring-opening polymerization technique was adapted to synthesize soft segment of (PCL-PEC-b-PDMS). Structure elucidation of the synthesized polymers is established by nuclear magnetic resonance spectroscopy. Sparingly, Al-mHA ornamented scaffolds exhibit tremendous improvement (175%) in the mechanical properties with promising antimicrobial activity against various human pathogens. After confirmation of high osteoconductivity, improved biodegradation, and excellent biocompatibility against osteoblast-like MG63 cells (in vitro), the scaffolds were implanted in rabbits as an animal model by subcutaneous and intraosseous (tibial) sites. Improved in vivo biocompatibilities, biodegradation, osteoconductivity, and the ability to provide an adequate biomimetic environment for biomineralization for GBR of the scaffolds (SPU and ornamented SPUs) have been found from the various histological sections. Early cartilage formation, endochondral ossification, and rapid bone healing at 4 weeks were found in the defects filled with Al-mHA ornamented

  1. Ectopic Bone Formation by Mesenchymal Stem Cells Derived from Human Term Placenta and the Decidua

    PubMed Central

    Gronthos, Stan; Manuelpillai, Ursula; Abumaree, Mohamed H.; Pertile, Mark D.; Brennecke, Shaun P.; Kalionis, Bill

    2015-01-01

    Mesenchymal stem cells (MSCs) are one of the most attractive cell types for cell-based bone tissue repair applications. Fetal-derived MSCs and maternal-derived MSCs have been isolated from chorionic villi of human term placenta and the decidua basalis attached to the placenta following delivery, respectively. Chorionic-derived MSCs (CMSCs) and decidua-derived MSCs (DMSCs) generated in this study met the MSCs criteria set by International Society of Cellular Therapy. These criteria include: (i) adherence to plastic; (ii) >90% expression of CD73, CD105, CD90, CD146, CD44 and CD166 combined with <5% expression of CD45, CD19 and HLA-DR; and (iii) ability to differentiate into osteogenic, adipogenic, and chondrogenic lineages. In vivo subcutaneous implantation into SCID mice showed that both bromo-deoxyuridine (BrdU)-labelled CMSCs and DMSCs when implanted together with hydroxyapatite/tricalcium phosphate particles were capable of forming ectopic bone at 8-weeks post-transplantation. Histological assessment showed expression of bone markers, osteopontin (OPN), osteocalcin (OCN), biglycan (BGN), bone sialoprotein (BSP), and also a marker of vasculature, alpha-smooth muscle actin (α-SMA). This study provides evidence to support CMSCs and DMSCs as cellular candidates with potent bone forming capacity. PMID:26484666

  2. Effects of Nano-hydroxyapatite/Poly(DL-lactic-co-glycolic acid) Microsphere-Based Composite Scaffolds on Repair of Bone Defects: Evaluating the Role of Nano-hydroxyapatite Content.

    PubMed

    He, Shu; Lin, Kai-Feng; Sun, Zhen; Song, Yue; Zhao, Yi-Nan; Wang, Zheng; Bi, Long; Liu, Jian

    2016-07-01

    The aim of the current study was to prepare microsphere-based composite scaffolds made of nano-hydroxyapatite (nHA)/poly (DL-lactic-co-glycolic acid) (PLGA) at different ratios and evaluate the effects of nHA on the characteristics of scaffolds for tissue engineering application. First, microsphere-based composite scaffolds made of two ratios of nHA/PLGA (nHA/PLGA = 20/80 and nHA/PLGA = 50/50) were prepared. Then, the effects of nHA on the wettability, mechanical strength, and degradation of scaffolds were investigated. Second, the biocompatibility and osteoinductivity were evaluated and compared by co-culture of scaffolds with bone marrow stromal stem cells (BMSCs). The results showed that the adhesion, proliferation, and osteogenic differentiation of BMSCs with nHA/PLGA (50/50) were better than those with nHA/PLGA (20/80). Finally, we implanted the scaffolds into femur bone defects in a rabbit model, then the capacity of guiding bone regeneration as well as the in vivo degradation were observed by micro-CT and histological examinations. After 4 weeks' implantation, there was no significant difference on the repair of bone defects. However, after 8 and 12 weeks' implantation, the nHA/PLGA (20/80) exhibited better bone formation than nHA/PLGA (50/50). These results suggested that a proper concentration of nHA in the nHA/PLGA composite should be taken into account when the composite scaffolds were prepared, which plays an important role in the biocompatibility, degradation rate and osteoconductivity. PMID:27378617

  3. Polydopamine-Templated Hydroxyapatite Reinforced Polycaprolactone Composite Nanofibers with Enhanced Cytocompatibility and Osteogenesis for Bone Tissue Engineering.

    PubMed

    Gao, Xiang; Song, Jinlin; Ji, Ping; Zhang, Xiaohong; Li, Xiaoman; Xu, Xiao; Wang, Mengke; Zhang, Siqi; Deng, Yi; Deng, Feng; Wei, Shicheng

    2016-02-10

    Nanohydroxyapatite (HA) synthesized by biomimetic strategy is a promising nanomaterial as bone substitute due to its physicochemical features similar to those of natural nanocrystal in bone tissue. Inspired by mussel adhesive chemistry, a novel nano-HA was synthesized in our work by employing polydopamine (pDA) as template under weak alkaline condition. Subsequently, the as-prepared pDA-templated HA (tHA) was introduced into polycaprolactone (PCL) matrix via coelectrospinning, and a bioactive tHA/PCL composite nanofiber scaffold was developed targeted at bone regeneration application. Our research showed that tHA reinforced PCL composite nanofibers exhibited favorable cytocompatibility at given concentration of tHA (0-10 w.t%). Compared to pure PCL and traditional nano-HA enriched PCL (HA/PCL) composite nanofibers, enhanced cell adhesion, spreading and proliferation of human mesenchymal stem cells (hMSCs) were observed on tHA/PCL composite nanofibers on account of the contribution of pDA present in tHA. More importantly, tHA nanoparticles exposed on the surface of composite nanofibers could further promote osteogenesis of hMSCs in vitro even in the absence of osteogenesis soluble inducing factors when compared to traditional HA/PCL scaffolds, which was supported by in vivo test as well according to the histological analysis. Overall, our study demonstrated that the developed tHA/PCL composite nanofibers with enhanced cytocompatibility and osteogenic capacity hold great potential as scaffolds for bone tissue engineering. PMID:26756224

  4. Comparative Study on the Efficacy of Anorganic Bovine Bone (Bio-Oss) and Nanocrystalline Hydroxyapatite (Ostim) in Maxillary Sinus Floor Augmentation

    PubMed Central

    Shirmohammadi, Adileh; Roshangar, Leila; Chitsazi, Mohammad Taghi; Pourabbas, Reza; Faramarzie, Masoumeh; Rahmanpour, Nasrin

    2014-01-01

    Purpose. The aim of this study was to evaluate the efficacy of anorganic bovine bone (Bio-Oss) in comparison with nanocrystalline hydroxyapatite (Ostim) in sinus floor augmentation. Methods. Ten patients aged 40–80 were selected. All the patients needed sinus floor augmentation due to insufficient bone for simultaneous implant placement. The patients underwent panoramic radiography and cone beam computed tomography (CBCT) prior to surgical procedure. After lifting the sinus membrane, Bio-Oss and Ostim are randomly grafted at one of the two sides. Biopsies were obtained from areas identified 5 months after the surgery and before implant placement and then were prepared for histological analysis. Statistical analysis was performed with nonparametric Wilcoxon signed-rank test for comparison of histological and radiological parameters between the two groups. Results. Histological findings revealed a significant increase in percentages of new bone in the Ostim group (P = 0.015). Furthermore, new bone density was greater with Ostim compared to Bio-Oss (P = 0.038); however, the difference in height increase after surgery did not reach statistical significance (P = 0.191). Conclusion. Despite the limitations of this trial, Ostim and Bio-Oss are useful biomaterials in sinus augmentation and Ostim seems to be even more effective in new bone formation.

  5. Comparative Study on the Efficacy of Anorganic Bovine Bone (Bio-Oss) and Nanocrystalline Hydroxyapatite (Ostim) in Maxillary Sinus Floor Augmentation.

    PubMed

    Shirmohammadi, Adileh; Roshangar, Leila; Chitsazi, Mohammad Taghi; Pourabbas, Reza; Faramarzie, Masoumeh; Rahmanpour, Nasrin

    2014-01-01

    Purpose. The aim of this study was to evaluate the efficacy of anorganic bovine bone (Bio-Oss) in comparison with nanocrystalline hydroxyapatite (Ostim) in sinus floor augmentation. Methods. Ten patients aged 40-80 were selected. All the patients needed sinus floor augmentation due to insufficient bone for simultaneous implant placement. The patients underwent panoramic radiography and cone beam computed tomography (CBCT) prior to surgical procedure. After lifting the sinus membrane, Bio-Oss and Ostim are randomly grafted at one of the two sides. Biopsies were obtained from areas identified 5 months after the surgery and before implant placement and then were prepared for histological analysis. Statistical analysis was performed with nonparametric Wilcoxon signed-rank test for comparison of histological and radiological parameters between the two groups. Results. Histological findings revealed a significant increase in percentages of new bone in the Ostim group (P = 0.015). Furthermore, new bone density was greater with Ostim compared to Bio-Oss (P = 0.038); however, the difference in height increase after surgery did not reach statistical significance (P = 0.191). Conclusion. Despite the limitations of this trial, Ostim and Bio-Oss are useful biomaterials in sinus augmentation and Ostim seems to be even more effective in new bone formation. PMID:27382621

  6. Analyzing the behavior of a porous nano-hydroxyapatite/polyamide 66 (n-HA/PA66) composite for healing of bone defects.

    PubMed

    Xiong, Yan; Ren, Cheng; Zhang, Bin; Yang, Hongsheng; Lang, Yun; Min, Li; Zhang, Wenli; Pei, Fuxing; Yan, Yonggang; Li, Hong; Mo, Anchun; Tu, Chongqi; Duan, Hong

    2014-01-01

    The aim of this study was to analyze the behavior of the porous nano-hydroxyapatite/polyamide 66 (n-HA/PA66) composite grafted for bone defect repair through a series of biological safety experiments, animal experiments, and a more than 5-year long clinical follow-up. The biological safety experiments, carried out in accordance with the Chinese Guo Biao and Tolerancing (GB/T)16886 and GB/T16175, revealed that porous n-HA/PA66 composite had no cytotoxicity, no sensitization effect, no pyrogenic reaction, and that its hemolysis rate was 0.59% (less than 5%). Rabbit models of tibia defects with grafted porous n-HA/PA66 composite were established. After 2 weeks, the experiment showed that osteogenesis was detected in the porous n-HA/PA66 composite; the density of new bone formation was similar to the surrounding host bone at 12 weeks. After 26 weeks, the artificial bone rebuilt to lamellar bone completely. In the clinical study, a retrospective review was carried out for 21 patients who underwent serial radiographic assessment after treatment with porous n-HA/PA66 composite grafts following bone tumor resection. All wounds healed to grade A. No postoperative infections, delayed deep infection, nonspecific inflammation, rejection, or fractures were encountered. At a mean follow-up of 5.3 years, the mean Musculoskeletal Tumor Society's (MSTS) 93 score was 29.3 points (range: 28-30 points) and mean radiopaque density ratio was 0.77±0.10. The radiologic analysis showed that porous n-HA/PA66 composite had been completely incorporated with the host bone about 1.5 years later. In conclusion, this study indicated that the porous n-HA/PA66 composite had biological safety, and good biocompatibility, osteoinduction, and osseointegration. Thus, the porous n-HA/PA66 composite is an ideal artificial bone substitute and worthy of promotion in the field. PMID:24531621

  7. Biosafety of the Novel Vancomycin-loaded Bone-like Hydroxyapatite/Poly-amino Acid Bony Scaffold

    PubMed Central

    Cao, Zhi-Dong; Jiang, Dian-Ming; Yan, Ling; Wu, Jun

    2016-01-01

    Background: Recently, local sustained-release antibiotics systems have been developed because they can increase local foci of concentrated antibiotics without increasing the plasma concentration, and thereby effectively decrease any systemic toxicity and side effects. A vancomycin-loaded bone-like hydroxyapatite/poly-amino acid (V-BHA/PAA) bony scaffold was successfully fabricated with vancomycin-loaded poly lactic-co-glycolic acid microspheres and BHA/PAA, which was demonstrated to exhibit both porosity and perfect biodegradability. The aim of this study was to systematically evaluate the biosafety of this novel scaffold by conducting toxicity tests in vitro and in vivo. Methods: According to the ISO rules for medical implant biosafety, for in vitro tests, the scaffold was incubated with L929 fibroblasts or rabbit noncoagulant blood, with simultaneous creation of positive control and negative control groups. The growth condition of L929 cells and hemolytic ratio were respectively evaluated after various incubation periods. For in vivo tests, a chronic osteomyelitis model involving the right proximal tibia of New Zealand white rabbits was established. After bacterial identification, the drug-loaded scaffold, drug-unloaded BHA/PAA, and poly (methyl methacrylate) were implanted, and a blank control group was also set up. Subsequently, the in vivo blood drug concentrations were measured, and the kidney and liver functions were evaluated. Results: In the in vitro tests, the cytotoxicity grades of V-BHA/PAA and BHA/PAA-based on the relative growth rate were all below 1. The hemolysis ratios of V-BHA/PAA and BHA/PAA were 2.27% and 1.42%, respectively, both below 5%. In the in vivo tests, the blood concentration of vancomycin after implantation of V-BHA/PAA was measured at far below its toxic concentration (60 mg/L), and the function and histomorphology of the liver and kidney were all normal. Conclusion: According to ISO standards, the V-BHA/PAA scaffold is considered to

  8. The effects of Paraloid B-72 and Butvar B-98 treatment and organic solvent removal on δ(13)C, δ(15)N, and δ(18)O values of collagen and hydroxyapatite in a modern bone.

    PubMed

    France, Christine A M; Giaccai, Jennifer A; Doney, Charlotte R

    2015-06-01

    Stable isotopes in bones are a powerful tool for diet, provenance, climate, and physiological reconstructions, but necessarily require well-preserved specimens unaltered by postmortem diagenesis or conservation practices. This study examines the effects of Paraloid B-72 and Butvar B-98, two common consolidants used in field and museum conservation, on δ(13)C, δ(15)N, and δ(18)O values from bone collagen and hydroxyapatite. The effects of solvent removal (100% acetone, 100% ethanol, 9:1 acetone:xylenes, 9:1 ethanol:xylenes) and drying methods (ambient air, vacuum, oven drying at 80°C) were also examined to determine if bones treated with these consolidants can successfully be cleaned and used for stable isotope analyses. Results show that introduction of Paraloid B-72 or Butvar B-98 in 100% acetone or 100% ethanol, respectively, with subsequent removal by the same solvents and drying at 80°C facilitates the most successful removal of consolidants and solvents. The δ(13)C values in collagen, δ(15)N in collagen, δ(18)O in hydroxyapatite phosphate, and δ(13)C in hydroxyapatite structural carbonate were unaltered by treatments with Paraloid or Butvar and subsequent solvent removal. The δ(18)O in hydroxyapatite structural carbonate showed nonsystematic variability when bones were treated with Paraloid and Butvar, which is hypothesized to be a result of hydroxyl exchange when bones are exposed to consolidants in solution. It is therefore recommended that δ(18)O in hydroxyapatite structural carbonate should not be used in stable isotope studies if bones have been treated with Paraloid or Butvar. PMID:25639211

  9. [Implantation of collagen coated hydroxyapatite particles. A clinical-histological study in humans].

    PubMed

    Sanz, M; Bascones, A; Kessler, A; García Nuñez, J; Newman, M G; Robertson, M A; Carranza, F A

    1989-05-01

    In this study, histologic behaviour of collagen coated hydroxylapatite particles implanted in human periodontal osseous defects has been analyzed. This material was surgically implanted in four patients, and reentry and block biopsies were carried out 4 and 6 months later. The histologic results demonstrate that this material is well tolerated by surrounding tissues, not eliciting an inflammatory reaction. At four months, the hydroxylapatite particles appear encapsulated by a very cellular connective tissue and at 6 months are found in direct contact with osteoid and mature bone. This material acts as a filler material, being fully biocompatible and stimulating an osseoconductive reaction of the adjacent alveolar bone. PMID:2637052

  10. Effects of Spaceflight on Bone: The Rat as an Animal Model for Human Bone Loss

    NASA Technical Reports Server (NTRS)

    Halloran, B.; Weider, T.; Morey-Holton, E.

    1999-01-01

    The loss of weight bearing during spaceflight results in osteopenia in humans. Decrements in bone mineral reach 3-10% after as little as 75-184 days in space. Loss of bone mineral during flight decreases bone strength and increases fracture risk. The mechanisms responsible for, and the factors contributing to, the changes in bone induced by spaceflight are poorly understood. The rat has been widely used as an animal model for human bone loss during spaceflight. Despite its potential usefulness, the results of bone studies performed in the rat in space have been inconsistent. In some flights bone formation is decreased and cancellous bone volume reduced, while in others no significant changes in bone occur. In June of 1996 Drs. T. Wronski, S. Miller and myself participated in a flight experiment (STS 78) to examine the effects of glucocorticoids on bone during weightlessness. Technically the 17 day flight experiment was flawless. The results, however, were surprising. Cancellous bone volume and osteoblast surface in the proximal tibial metaphysis were the same in flight and ground-based control rats. Normal levels of cancellous bone mass and bone formation were also detected in the lumbar vertebrae and femoral neck of flight rats. Furthermore, periosteal bone formation rate was found to be identical in flight and ground-based control rats. Spaceflight had little or no effect on bone metabolism! These results prompted us to carefully review the changes in bone observed in, and the flight conditions of previous spaceflight missions.

  11. Influence of plasma spraying deposition process on optical properties of hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Belka, Radosław; Kowalski, Szymon; Żórawski, Wojciech; Suchańska, Małgorzata

    2015-09-01

    Hydroxyapatite (HAp) is a well-known bioceramic, nonorganic material of the bones of the vertebrate responsible for their mechanical durability. In human bones it occupies 60-80 % of the volume depending on a number of factors. Synthetic HAp is valued in bone endoprosthetic to its high biocompatibility. It is widely used to fill cavities of bone and as the coating of bone implants to increase their biocompatibility and adhesion to bone surface. In this paper a diffuse reflectance spectra of plasma-spraying deposited hydroxyapatite were presented and compared with pure powder samples. Optical band gap were estimated basing on Kubelka-Munk functions and Tauc plot extrapolation. We found that deposition process affects the value of band gap.

  12. Mechanistic fracture criteria for the failure of human cortical bone

    SciTech Connect

    Nalla, Ravi K.; Kinney, John H.; Ritchie, Robert O.

    2002-12-13

    A mechanistic understanding of fracture in human bone is critical to predicting fracture risk associated with age and disease. Despite extensive work, a mechanistic framework for describing how the underlying microstructure affects the failure mode in bone is lacking.

  13. Enhancing the bioactivity of Poly(lactic-co-glycolic acid) scaffold with a nano-hydroxyapatite coating for the treatment of segmental bone defect in a rabbit model

    PubMed Central

    Wang, De-Xin; He, Yao; Bi, Long; Qu, Ze-Hua; Zou, Ji-Wei; Pan, Zhen; Fan, Jun-Jun; Chen, Liang; Dong, Xin; Liu, Xiang-Nan; Pei, Guo-Xian; Ding, Jian-Dong

    2013-01-01

    Purpose Poly(lactic-co-glycolic acid) (PLGA) is excellent as a scaffolding matrix due to feasibility of processing and tunable biodegradability, yet the virgin scaffolds lack osteoconduction and osteoinduction. In this study, nano-hydroxyapatite (nHA) was coated on the interior surfaces of PLGA scaffolds in order to facilitate in vivo bone defect restoration using biomimetic ceramics while keeping the polyester skeleton of the scaffolds. Methods PLGA porous scaffolds were prepared and surface modification was carried out by incubation in modified simulated body fluids. The nHA coated PLGA scaffolds were compared to the virgin PLGA scaffolds both in vitro and in vivo. Viability and proliferation rate of bone marrow stromal cells of rabbits were examined. The constructs of scaffolds and autogenous bone marrow stromal cells were implanted into the segmental bone defect in the rabbit model, and the bone regeneration effects were observed. Results In contrast to the relative smooth pore surface of the virgin PLGA scaffold, a biomimetic hierarchical nanostructure was found on the surface of the interior pores of the nHA coated PLGA scaffolds by scanning electron microscopy. Both the viability and proliferation rate of the cells seeded in nHA coated PLGA scaffolds were higher than those in PLGA scaffolds. For bone defect repairing, the radius defects had, after 12 weeks implantation of nHA coated PLGA scaffolds, completely recuperated with significantly better bone formation than in the group of virgin PLGA scaffolds, as shown by X-ray, Micro-computerized tomography and histological examinations. Conclusion nHA coating on the interior pore surfaces can significantly improve the bioactivity of PLGA porous scaffolds. PMID:23690683

  14. MicroRNA-26a-modified adipose-derived stem cells incorporated with a porous hydroxyapatite scaffold improve the repair of bone defects

    PubMed Central

    WANG, ZHENLIN; ZHANG, DAWEI; HU, ZHIQIANG; CHENG, JIWEI; ZHUO, CHUANMENG; FANG, XIANCONG; XING, YONGMING

    2015-01-01

    Tissue-engineered bone substitutes are frequently used to repair bone defects. Adipose-derived stem cells (ASCs) are a promising source of cells for repairing bone tissue, however, insufficient osteogenic potency remains the main obstacle for their application. The present study aimed to enhance the osteogenic potency of ASCs by transfection of microRNA (miR)-26a, a novel osteogenic and angiogenic promoting miRNA. An inverted fluorescence microscope was used to observe transfection efficiency, while a scanning electron microscope was used to detect morphological alterations. Cell proliferation was monitored continuously for 7 days using a Cell Counting kit-8 assay. Osteogenic differentiation was determined by reverse transcription quantitative polymerase chain reaction, alkaline phosphatase (ALP) staining, collagen secretion and extracellular matrix (ECM) mineralization. ASCs were incorporated with a porous hydroxyapatite (HA) scaffold to create a novel tissue-engineered bone substitute and inserted into the critical tibia defect of rats. New bone formation was evaluated by hematoxylin and eosin and Masson's trichrome staining. The results demonstrated that miR-26a was successfully delivered into the cytoplasm, while the morphology and proliferation of ASCs were not significantly altered. Osteogenic-associated genes were markedly upregulated and ALP production, collagen secretion and ECM mineralization were all increased following transfection of miR-26a. Histological evaluation demonstrated that the modified cells accompanied with a porous HA scaffold markedly promoted new bone formation within the defective area. In conclusion, miR-26a transfection significantly improved the osteogenic potency of ASCs suggesting that modified ASCs incorporated with a HA scaffold may be used as a potential bone substitute. PMID:25997460

  15. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells

    PubMed Central

    Wu, Xiaonan; Miao, Leiying; Yao, Yingfang; Wu, Wenlei; Liu, Yu; Chen, Xiaofeng; Sun, Weibin

    2014-01-01

    Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(ε-caprolactone) (COL/PCL) and type I collagen/poly(ε-caprolactone)/nanoscale hydroxyapatite (COL/PCL/nHA) with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF) immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then extended gradually with stretched filopodia, indicating an ability to fill the fiber pores. A Cell Counting Kit-8 assay showed that both scaffolds supported cell proliferation. However, real-time quantitative polymerase chain reaction analysis showed that expression of the bone-related markers, alkaline phosphatase and osteocalcin, was upregulated only on the COL/PCL/nHA-SBF scaffold, indicating that this scaffold had the ability to induce osteogenic differentiation of periodontal ligament cells. In this study, COL/PCL/nHA-SBF produced by electrospinning followed by biomimetic mineralization had combined electrospun fibers with nHA in it. This scaffold has

  16. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells.

    PubMed

    Wu, Xiaonan; Miao, Leiying; Yao, Yingfang; Wu, Wenlei; Liu, Yu; Chen, Xiaofeng; Sun, Weibin

    2014-01-01

    Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(ε-caprolactone) (COL/PCL) and type I collagen/poly(ε-caprolactone)/nanoscale hydroxyapatite (COL/PCL/nHA) with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF) immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then extended gradually with stretched filopodia, indicating an ability to fill the fiber pores. A Cell Counting Kit-8 assay showed that both scaffolds supported cell proliferation. However, real-time quantitative polymerase chain reaction analysis showed that expression of the bone-related markers, alkaline phosphatase and osteocalcin, was upregulated only on the COL/PCL/nHA-SBF scaffold, indicating that this scaffold had the ability to induce osteogenic differentiation of periodontal ligament cells. In this study, COL/PCL/nHA-SBF produced by electrospinning followed by biomimetic mineralization had combined electrospun fibers with nHA in it. This scaffold has

  17. Aesthetic recovery of alveolar atrophy following autogenous onlay bone grafting using interconnected porous hydroxyapatite ceramics (IP-CHA) and resorbable poly-L-lactic/polyglycolic acid screws: case report

    PubMed Central

    2014-01-01

    Background Onlay bone grafting techniques have some problems related to the limited volume of autogenous grafted bone and need for surgery to remove bone fixing screws. Here, we report a case of horizontal alveolar ridge atrophy following resection of a maxillary bone cyst, in which autogenous onlay bone grafting with interconnected porous hydroxyapatite ceramics (IP-CHA) and bioresorbable poly-L-lactic/polyglycolic acid (PLLA-PGA) screws was utilized. Case presentation A 51-year-old man had aesthetic complications related to alveolar atrophy following maxillary bone cyst extraction. We performed onlay grafting for aesthetic alveolar bone recovery using IP-CHA to provide adequate horizontal bone volume and PLLA-PGA screws for bone fixing to avoid later damage to host bone during surgical removal. During the operation, an autogenous cortical bone block was collected from the ramus mandibular and fixed to the alveolar ridge with PLLA-PGA screws, then the gap between the bone block and recipient bone was filled with a granular type of IP-CHA. Post-surgery orthopantomograph and CT scan findings showed no abnormal resorption of the grafted bone, and increased radiopacity, which indicated new bone formation in the area implanted with IP-CHA. Conclusion Our results show that IP-CHA and resorbable PLLA-PGA screws are useful materials for autogenous onlay bone grafting. PMID:24889647

  18. Development and characterization of an injectable cement of nano calcium-deficient hydroxyapatite/multi(amino acid) copolymer/calcium sulfate hemihydrate for bone repair

    PubMed Central

    Qi, Xiaotong; Li, Hong; Qiao, Bo; Li, Weichao; Hao, Xinyan; Wu, Jun; Su, Bao; Jiang, Dianming

    2013-01-01

    A novel injectable bone cement was developed by integration of nano calcium-deficient hydroxyapatite/multi(amino acid) copolymer (n-CDHA/MAC) and calcium sulfate hemihydrate (CSH; CaSO4 · 1/2H2O). The structure, setting time, and compressive strength of the cement were investigated. The results showed that the cement with a liquid to powder ratio of 0.8 mL/g exhibited good injectability and appropriate setting time and mechanical properties. In vitro cell studies indicated that MC3T3-E1 cells cultured on the n-CDHA/MAC/CSH composite spread well and showed a good proliferation state. The alkaline phosphatase activity of the MC3T3-E1 cells cultured on the n-CDHA/MAC/CSH composite was significantly higher than that of the cells on pure CSH at 4 and 7 days of culture. The n-CDHA/MAC/CSH cement was implanted into critical size defects of the femoral condyle in rabbits to evaluate its biocompatibility and osteogenesis in vivo. Radiological and histological results indicated that introduction of the n-CDHA/MAC into CSH enhanced new bone formation, and the n-CDHA/MAC/CSH cement exhibited good biocompatibility and degradability. In conclusion, the injectable n-CDHA/MAC/CSH composite cement has a significant clinical advantage over pure CSH cement, and may be a promising bone graft substitute for the treatment of bone defects. PMID:24293996

  19. Development and characterization of an injectable cement of nano calcium-deficient hydroxyapatite/multi(amino acid) copolymer/calcium sulfate hemihydrate for bone repair.

    PubMed

    Qi, Xiaotong; Li, Hong; Qiao, Bo; Li, Weichao; Hao, Xinyan; Wu, Jun; Su, Bao; Jiang, Dianming

    2013-01-01

    A novel injectable bone cement was developed by integration of nano calcium-deficient hydroxyapatite/multi(amino acid) copolymer (n-CDHA/MAC) and calcium sulfate hemihydrate (CSH; CaSO4 · 1/2H2O). The structure, setting time, and compressive strength of the cement were investigated. The results showed that the cement with a liquid to powder ratio of 0.8 mL/g exhibited good injectability and appropriate setting time and mechanical properties. In vitro cell studies indicated that MC3T3-E1 cells cultured on the n-CDHA/MAC/CSH composite spread well and showed a good proliferation state. The alkaline phosphatase activity of the MC3T3-E1 cells cultured on the n-CDHA/MAC/CSH composite was significantly higher than that of the cells on pure CSH at 4 and 7 days of culture. The n-CDHA/MAC/CSH cement was implanted into critical size defects of the femoral condyle in rabbits to evaluate its biocompatibility and osteogenesis in vivo. Radiological and histological results indicated that introduction of the n-CDHA/MAC into CSH enhanced new bone formation, and the n-CDHA/MAC/CSH cement exhibited good biocompatibility and degradability. In conclusion, the injectable n-CDHA/MAC/CSH composite cement has a significant clinical advantage over pure CSH cement, and may be a promising bone graft substitute for the treatment of bone defects. PMID:24293996

  20. Development of bone and cartilage in tissue-engineered human middle phalanx models.

    PubMed

    Wada, Yoshitaka; Enjo, Mitsuhiro; Isogai, Noritaka; Jacquet, Robin; Lowder, Elizabeth; Landis, William J

    2009-12-01

    Human middle phalanges were tissue-engineered with midshaft scaffolds of poly(L-lactide-epsilon-caprolactone) [P(LA-CL)], hydroxyapatite-P(LA-CL), or beta-tricalcium phosphate-P(LA-CL) and end plate scaffolds of bovine chondrocyte-seeded polyglycolic acid. Midshafts were either wrapped with bovine periosteum or left uncovered. Constructs implanted in nude mice for up to 20 weeks were examined for cartilage and bone development as well as gene expression and protein secretion, which are important in extracellular matrix (ECM) formation and mineralization. Harvested 10- and 20-week constructs without periosteum maintained end plate cartilage but no growth plate formation. They also consisted of chondrocytes secreting type II collagen and proteoglycan, and they were composed of midshaft regions devoid of bone. In all periosteum-wrapped constructs at like times, end plate scaffolds held chondrocytes elaborating type II collagen and proteoglycan and cartilage growth plates resembling normal tissue. Chondrocyte gene expression of type II collagen, aggrecan, and bone sialoprotein varied depending on midshaft composition, presence of periosteum, and length of implantation time. Periosteum produced additional cells, ECM, and mineral formation within the different midshaft scaffolds. Periosteum thus induces midshaft development and mediates chondrocyte gene expression and growth plate formation in cartilage regions of phalanges. This work is important for understanding developmental principles of tissue-engineered phalanges and by extension those of normal growth plate cartilage and bone. PMID:19527181

  1. Development of Bone and Cartilage in Tissue-Engineered Human Middle Phalanx Models

    PubMed Central

    Wada, Yoshitaka; Enjo, Mitsuhiro; Isogai, Noritaka; Jacquet, Robin; Lowder, Elizabeth

    2009-01-01

    Human middle phalanges were tissue-engineered with midshaft scaffolds of poly(L-lactide-ɛ-caprolactone) [P(LA-CL)], hydroxyapatite-P(LA-CL), or β-tricalcium phosphate-P(LA-CL) and end plate scaffolds of bovine chondrocyte-seeded polyglycolic acid. Midshafts were either wrapped with bovine periosteum or left uncovered. Constructs implanted in nude mice for up to 20 weeks were examined for cartilage and bone development as well as gene expression and protein secretion, which are important in extracellular matrix (ECM) formation and mineralization. Harvested 10- and 20-week constructs without periosteum maintained end plate cartilage but no growth plate formation. They also consisted of chondrocytes secreting type II collagen and proteoglycan, and they were composed of midshaft regions devoid of bone. In all periosteum-wrapped constructs at like times, end plate scaffolds held chondrocytes elaborating type II collagen and proteoglycan and cartilage growth plates resembling normal tissue. Chondrocyte gene expression of type II collagen, aggrecan, and bone sialoprotein varied depending on midshaft composition, presence of periosteum, and length of implantation time. Periosteum produced additional cells, ECM, and mineral formation within the different midshaft scaffolds. Periosteum thus induces midshaft development and mediates chondrocyte gene expression and growth plate formation in cartilage regions of phalanges. This work is important for understanding developmental principles of tissue-engineered phalanges and by extension those of normal growth plate cartilage and bone. PMID:19527181

  2. Clinical and radiographic evaluation of Nano-crystalline hydroxyapatite bone graft (Sybograf®) in combination with bioresorbable collagen membrane (Periocol®) in periodontal intrabony defects

    PubMed Central

    Singh, Vijendra P.; Nayak, Dilip G.; Uppoor, Ashita S.; Shah, Dipen

    2012-01-01

    Background: Nanosized ceramics may represent a promising class of bone graft substitutes due to their improved osseointegrative properties. Nanocrystalline Hydroxyapatite (NcHA) bind to bone and stimulate bone healing by stimulation of osteoblast activity. The present study aims to explore the clinical and radiographical outcome of NcHA bonegraft (Sybograf®) with collagen membrane (Periocol®), in comparison with open flap debridement (OFD), in the treatment of intrabony periodontal defects. Materials and Methods: A parallel-group, randomized, controlled clinical trial was designed to conduct the study. Eighteen intrabony defects in 14 systemically healthy patients aged between 25 to 65 years were randomly assigned to test and control group. The plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL), and gingival recession (REC) were recorded at baseline, and were reevaluated at 6 months. In addition to this, radiographic bone fill was assessed using digital software. At the test site, NcHA bone graft and collagen membrane was placed, whereas at the control site, only OFD was done. Recall appointments were made at 7 days, 30 days, and then at 3 months and 6 months. Results: The data were subjected to statistical analysis using the Mann-Whitney ‘U’ Test and Wilcoxon signed rank sum test. In the control group, the mean reduction of PPD was 3.22±1.09 mm (P=0.007) and CAL gain was 2.77±1.09 mm (P=0.007). In the test group, the mean PPD reduction of 4.33±0.5 mm (P=0.006) and mean gain in CAL was 3.77±0.66 mm (P=0.006) at 6 months. The mean increase in REC was 0.55±0.72 mm (P=0.025) in test, and 0.44±0.52 mm (P=0.046) in control group. The mean gain in radiographic defect fill was 2.07±0.67 mm (P=0.008) in test and 0.91±0.21 mm (P=0.007) in control group. Conclusion: The nanocrystalline hydroxyapatite bone graft in combination with collagen membrane demonstrated clinical advantages beyond that achieved by OFD alone. PMID

  3. A glass-reinforced hydroxyapatite and surgical-grade calcium sulfate for bone regeneration: In vivo biological behavior in a sheep model.

    PubMed

    Cortez, Paulo Pegado; Silva, Marta Alves; Santos, Marta; Armada-da-Silva, Paulo; Afonso, Amrico; Lopes, Maria A; Santos, Jose Domingos; Maurício, Ana Colette

    2012-08-01

    A glass-reinforced hydroxyapatite (HA) composite (Bonelike®) was developed for bone grafting. This biomaterial is composed of a modified HA matrix with α- and β-tricalcium phosphate secondary phases, resulting in higher solubility than single HA type of materials. Several in vitro and in vivo studies demonstrated that Bonelike® has a highly bioactive behavior, which was also confirmed by employing granular forms of this biomaterial in orthopedics and dental applications. However, a fast consolidation vehicle was needed to promote the fixation of Bonelike® granules if applied in larger defects or in unstable sites. Surgical-grade calcium sulfate (CS), which is widely recognized as a well-tolerated and inexpensive bone graft material, was the chosen vehicle to improve the handling characteristics of Bonelike® as it can be used in the form of a powder that is mixed with a liquid to form a paste that sets in situ. After application in non-critical monocortical defects in sheep, histological, and scanning electron microscopy evaluations demonstrated that Bonelike® associated to CS functioned as a very satisfactory scaffold for bone regeneration as it achieved synchronization of the ingrowing bone with biomaterial resorption and subsequent preservation of the bone graft initial volume. Therefore, our results indicate that CS is an effective vehicle for Bonelike® granules as it facilitates their application and does not interfere with their proven highly osteoconductive properties. In the opposite way, the incorporation of Bonelike® improves the bone regeneration capabilities of CS. PMID:21602251

  4. A novel strontium(II)-modified calcium phosphate bone cement stimulates human-bone-marrow-derived mesenchymal stem cell proliferation and osteogenic differentiation in vitro.

    PubMed

    Schumacher, M; Lode, A; Helth, A; Gelinsky, M

    2013-12-01

    In the present study, the in vitro effects of novel strontium-modified calcium phosphate bone cements (SrCPCs), prepared using two different approaches on human-bone-marrow-derived mesenchymal stem cells (hMSCs), were evaluated. Strontium ions, known to stimulate bone formation and therefore already used in systemic osteoporosis therapy, were incorporated into a hydroxyapatite-forming calcium phosphate bone cement via two simple approaches: incorporation of strontium carbonate crystals and substitution of Ca(2+) by Sr(2+) ions during cement setting. All modified cements released 0.03-0.07 mM Sr(2+) under in vitro conditions, concentrations that were shown not to impair the proliferation or osteogenic differentiation of hMSCs. Furthermore, strontium modification led to a reduced medium acidification and Ca(2+) depletion in comparison to the standard calcium phosphate cement. In indirect and direct cell culture experiments with the novel SrCPCs significantly enhanced cell proliferation and differentiation were observed. In conclusion, the SrCPCs described here could be beneficial for the local treatment of defects, especially in the osteoporotic bone. PMID:23917042

  5. Characterization of hydroxyapatite coating by pulse laser deposition technique on stainless steel 316 L by varying laser energy

    NASA Astrophysics Data System (ADS)

    Khandelwal, Himanshu; Singh, Gurbhinder; Agrawal, Khelendra; Prakash, Satya; Agarwal, R. D.

    2013-01-01

    Hydroxyapatite is an attractive biomaterial mainly used in bone and tooth implants because it closely resembles human tooth and bone mineral and has proven to be biologically compatible with these tissues. In spite of this advantage of hydroxyapatite it has also certain limitation like inferior mechanical properties which do not make it suitable for long term load bearing applications; hence a lot of research is going on in the development of hydroxyapatite coating over various metallic implants. These metallic implants have good biocompatibility and mechanical properties. The aim of the present work is to deposit hydroxyapatite coating over stainless steel grade 316 L by pulse laser deposition technique by varying laser energy. To know the effect of this variation, the coatings were than characterized in detail by X-ray diffraction, finite emission-scanning electron microscope, atomic force microscope and energy dispersive X-ray spectroscopy.

  6. In Vitro and In Vivo Evaluations of Nano-Hydroxyapatite/Polyamide 66/Glass Fibre (n-HA/PA66/GF) as a Novel Bioactive Bone Screw

    PubMed Central

    Su, Bao; Peng, Xiaohua; Jiang, Dianming; Wu, Jun; Qiao, Bo; Li, Weichao; Qi, Xiaotong

    2013-01-01

    In this study, we prepared nano-hydroxyapatite/polyamide 66/glass fibre (n-HA/PA66/GF) bioactive bone screws. The microstructure, morphology and coating of the screws were characterised, and the adhesion, proliferation and viability of MC3T3-E1 cells on n-HA/PA66/GF scaffolds were determined using scanning electron microscope, CCK-8 assays and cellular immunofluorescence analysis. The results confirmed that n-HA/PA66/GF scaffolds were biocompatible and had no negative effect on MC3T3-E1 cells in vitro. To investigate the in vivo biocompatibility, internal fixation properties and osteogenesis of the bioactive screws, both n-HA/PA66/GF screws and metallic screws were used to repair intercondylar femur fractures in dogs. General photography, CT examination, micro-CT examination, histological staining and biomechanical assays were performed at 4, 8, 12 and 24 weeks after operation. The n-HA/PA66/GF screws exhibited good biocompatibility, high mechanical strength and extensive osteogenesis in the host bone. Moreover, 24 weeks after implantation, the maximum push-out load of the bioactive screws was greater than that of the metallic screws. As shown by their good cytocompatibility, excellent biomechanical strength and fast formation and ingrowth of new bone, n-HA/PA66/GF screws are thus suitable for orthopaedic clinical applications. PMID:23861888

  7. A specific groove design for individualized healing in a canine partial sternal defect model by a polycaprolactone/hydroxyapatite scaffold coated with bone marrow stromal cells.

    PubMed

    Xuan, Yiwen; Tang, Hua; Wu, Bin; Ding, Xinyu; Lu, Zhongyuan; Li, Wei; Xu, Zhifei

    2014-10-01

    The reconstruction of sternal defects remains clinically challenging for thoracic surgeons. Here we aimed to explore the individualized reconstruction of partial sternal defects with new biodegradable material in a large animal model. We used the fused deposition modeling (FDM) technique to manufacture polycaprolactone/hydroxyapatite (PCL/HA) tissue scaffolds with individualized grooves to repair the sternal defect. The defects were surgically created in a sternocostal joint of eighteen Beagle dogs. The animals were separated into three groups (n = 6): Blank group, PCL/HA group, and PCL/HA/BMSCs group. Radiographic examination, histological, and histomorphometric analyses were performed to evaluate the result. In the blank group, the defect site couldn't maintain its original integrity due to no bone union. In the PCL/HA group and PCL/HA/BMSCs group, it was observed that the scaffolds retained their shapes without significant degradation at 12 weeks. Both groups could observe new bone-union by radiographic and histological examination. And PCL/HA/BMSCs would be more mineralized tissue area at implant sites (p < 0.05). These results reveal that using the FDM technique to manufacture the PCL/HA scaffolds with specific grooves could repair the sternal defect satisfactorily. Furthermore the scaffolds with BMSCs-seeded could enhance the amount of bone ingrowth and seemed to be more promising. PMID:24142768

  8. Defect-Related Luminescent Hydroxyapatite-Enhanced Osteogenic Differentiation of Bone Mesenchymal Stem Cells Via an ATP-Induced cAMP/PKA Pathway.

    PubMed

    Wang, Chao; Liu, Dandan; Zhang, Cuimiao; Sun, Jiadong; Feng, Weipei; Liang, Xing-Jie; Wang, Shuxiang; Zhang, Jinchao

    2016-05-11

    Novel defect-related hydroxyapatite (DHAP), which combines the advantages of HAP and defect-related luminescence, has the potential application in tissue engineering and biomedical area, because of its excellent capability of monitoring the osteogenic differentiation and material biodegradation. Although the extracellular mechanism of DHAP minerals and PO4(3-) functioning in osteogenic differentiation has been widely studied, the intracellular molecular mechanism through which PO4(3-) mediates osteogenesis of bone mesenchymal stem cells (BMSCs) is not clear. We examined a previously unknown molecular mechanism through which PO4(3-) promoted osteogenesis of BMSCs with an emphasis on adenosine-triphosphate (ATP)-induced cAMP/PKA pathway. Our studies showed that DHAP could be uptaken into lysosome, in which PO4(3-) was released from DHAP, because of the acid environment of lysosome. The released PO4(3-) interacted with ADP to form ATP, and then degraded into adenosine, an ATP metabolite, which interacted with A2b adenosine receptor to activate the cAMP/PKA pathway, resulting in the high expression of osteogenesis-related genes, such as Runx2, BMP-2, and OCN. These findings first revealed the function of ATP-metabolism in bone physiological homeostasis, which may be developed to cure bone metabolic diseases. PMID:27088570

  9. Parameters in three-dimensional osteospheroids of telomerized human mesenchymal (stromal) stem cells grown on osteoconductive scaffolds that predict in vivo bone-forming potential.

    PubMed

    Burns, Jorge S; Rasmussen, Pernille L; Larsen, Kenneth H; Schrøder, Henrik Daa; Kassem, Moustapha

    2010-07-01

    Osteoblastic differentiation of human mesenchymal stem cells (hMSC) in monolayer culture is artefactual, lacking an organized bone-like matrix. We present a highly reproducible microwell protocol generating three-dimensional ex vivo multicellular aggregates of telomerized hMSC (hMSC-telomerase reverse transcriptase (TERT)) with improved mimicry of in vivo tissue-engineered bone. In osteogenic induction medium the hMSC were transitioned with time-dependent specification toward the osteoblastic lineage characterized by production of alkaline phosphatase, type I collagen, osteonectin, and osteocalcin. Introducing a 1-2 mm(3) crystalline hydroxyapatite/beta-tricalcium phosphate scaffold generated osteospheroids with upregulated gene expression of transcription factors RUNX2/CBFA1, Msx-2, and Dlx-5. An organized lamellar bone-like collagen matrix, evident by birefringence of polarized light, was deposited in the scaffold concavities. Here, mature osteoblasts stained positively for differentiated osteoblast markers TAZ, biglycan, osteocalcin, and phospho-AKT. Quantification of collagen birefringence and relatively high expression of genes for matrix proteins, including type I collagen, biglycan, decorin, lumican, elastin, microfibrillar-associated proteins (MFAP2 and MFAP5), periostin, and tetranectin, in vitro correlated predictively with in vivo bone formation. The three-dimensional hMSC-TERT/hydroxyapatite-tricalcium phosphate osteospheroid cultures in osteogenic induction medium recapitulated many characteristics of in vivo bone formation, providing a highly reproducible and resourceful platform for improved in vitro modeling of osteogenesis and refinement of bone tissue engineering. PMID:20196644

  10. A human salivary protein which promotes adhesion of Streptococcus mutans serotype c strains to hydroxyapatite.

    PubMed Central

    Kishimoto, E; Hay, D I; Gibbons, R J

    1989-01-01

    The aim of this study was to investigate the nature of one of the factors in human submandibular-sublingual (SMSL) saliva which promotes the adhesion of Streptococcus mutans serotype c strains to hydroxyapatite (HA) surfaces. Gel filtration chromatography of SMSL saliva on Trisacryl GF2000 gave a void volume peak which contained the major fraction of adhesion-promoting activity for S. mutans JBP to HA. Maximum adhesion-promoting activity, however, eluted slightly later than the maximum 220-nm absorbance of the void volume peak. Gel filtration of the void volume material after treatment with sodium dodecyl sulfate (SDS) gave an early-eluting larger peak followed by a smaller peak with which the adhesion-promoting activity was associated. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed the presence of relatively slowly migrating material associated with the larger inactive peak, presumably mucin, and a faster-migrating band(s) associated with the smaller active peak. SDS-PAGE indicated molecular weights in the range of 300,000 to 350,000 by extrapolation from size standards. Comparison of SMSL from five individuals showed the presence of single bands or double bands associated with adhesion-promoting activity, indicating genetic polymorphism. The active material did not resemble either secretory immunoglobulin A, based on SDS-PAGE and immunoassay, or fibronectin, based on SDS-PAGE, and also differed in molecular weight from salivary mucins and salivary constituents previously reported to promote aggregation of certain oral bacteria, but a relationship to these materials cannot be excluded. This adhesion-promoting material may play a significant role in the initial colonization of tooth surfaces by S. mutans strains. Images PMID:2807544

  11. The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part II: milled bone powders, reprecipitated hydroxyapatite and the potential uses of these materials.

    PubMed

    Johnson, G S; Mucalo, M R; Lorier, M A; Gieland, U; Mucha, H

    2000-11-01

    Further studies on the processing and use of animal-bone-derived calcium phosphate materials in biomedical applications are presented. Bone powders sourced either from the direct crushing and milling of bovine, ovine and cervine bone or after being subjected to defatting and acid digestion/NaOH reprecipitation and sodium hypochlorite hydrogen peroxide treatment of animal bones were characterized using Fourier transform infra-red (FTIR) spectroscopy, 13C solid state magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy, atomic absorption (AA) and inductively coupled plasma (ICP) spectrometric techniques. Bone powders were trialled for their potential use as a substrate for phosphine coupling and enzyme immobilization as well as a feedstock powder for plasma spraying on titanium metal substrates. Results indicated that enzyme immobilization by phosphine coupling could be successfully achieved on milled cervine bone with the immobilized enzyme retaining some activity. It was found that the presence of impurities normally carried down with the processing of the bone materials (viz., fat and collagen) played an important role in influencing the adsorbency and reactivity of the powders. Plasma spraying studies using reprecipitated bovine-derived powders produced highly adherent coatings on titanium metal, the composition of which was mostly hydroxyapatite (Ca10(PO4)6(OH)2) with low levels of alpha-tricalcium phosphate (alpha-Ca3(PO4)2) and tetracalcium phosphate (Ca4P2O9) also detected. In general, animal derived calcium phosphate materials constitute a potentially cheaper source of calcium phosphate materials for biomedical applications and make use of a largely under-utilized resource from abattoir wastes. PMID:15348079

  12. Hydroxyapatite formation on titania-based materials in a solution mimicking body fluid: Effects of manganese and iron addition in anatase.

    PubMed

    Shin, Euisup; Kim, Ill Yong; Cho, Sung Baek; Ohtsuki, Chikara

    2015-03-01

    Hydroxyapatite formation on the surfaces of implanted materials plays an important role in osteoconduction of bone substitutes in bone tissues. Titania hydrogels are known to instigate hydroxyapatite formation in a solution mimicking human blood plasma. To date, the relationship between the surface characteristics of titania and hydroxyapatite formation on its surface remains unclear. In this study, titania powders with varying surface characteristics were prepared by addition of manganese or iron to examine hydroxyapatite formation in a type of simulated body fluid (Kokubo solution). Hydroxyapatite formation was monitored by observation of deposited particles with scale-like morphology on the prepared titania powders. The effect of the titania surface characteristics, i.e., crystal structure, zeta potential, hydroxy group content, and specific surface area, on hydroxyapatite formation was examined. Hydroxyapatite formation was observed on the surface of titania powders that were primarily anatase, and featured a negative zeta potential and low specific surface areas irrespective of the hydroxy group content. High specific surface areas inhibited the formation of hydroxyapatite because calcium and phosphate ions were mostly consumed by adsorption on the titania surface. Thus, these surface characteristics of titania determine its osteoconductivity following exposure to body fluid. PMID:25579924

  13. Application of synchrotron radiation computed microtomography for quantification of bone microstructure in human and rat bones

    NASA Astrophysics Data System (ADS)

    Nogueira, Liebert Parreiras; Barroso, Regina Cély; de Almeida, André Pereira; Braz, Delson; de Almeida, Carlos Eduardo; de Andrade, Cherley Borba; Tromba, Giuliana

    2012-05-01

    This work aims to evaluate histomorphometric quantification by synchrotron radiation computed microto-mography in bones of human and rat specimens. Bones specimens are classified as normal and pathological (for human samples) and irradiated and non-irradiated samples (for rat ones). Human bones are specimens which were affected by some injury, or not. Rat bones are specimens which were irradiated, simulating radiotherapy procedures, or not. Images were obtained on SYRMEP beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. The system generated 14 μm tomographic images. The quantification of bone structures were performed directly by the 3D rendered images using a home-made software. Resolution yielded was excellent what facilitate quantification of bone microstructures.

  14. Application of synchrotron radiation computed microtomography for quantification of bone microstructure in human and rat bones

    SciTech Connect

    Parreiras Nogueira, Liebert; Barroso, Regina Cely; Pereira de Almeida, Andre; Braz, Delson; Almeida, Carlos Eduardo de; Borba de Andrade, Cherley; Tromba, Giuliana

    2012-05-17

    This work aims to evaluate histomorphometric quantification by synchrotron radiation computed microto-mography in bones of human and rat specimens. Bones specimens are classified as normal and pathological (for human samples) and irradiated and non-irradiated samples (for rat ones). Human bones are specimens which were affected by some injury, or not. Rat bones are specimens which were irradiated, simulating radiotherapy procedures, or not. Images were obtained on SYRMEP beamline at the Elettra Synchrotron Laboratory in Trieste, Italy. The system generated 14 {mu}m tomographic images. The quantification of bone structures were performed directly by the 3D rendered images using a home-made software. Resolution yielded was excellent what facilitate quantification of bone microstructures.

  15. Complex effect of hydroxyapatite nanoparticles on the differentiation and functional activity of human pre-osteoclastic cells.

    PubMed

    Costa-Rodrigues, João; Silva, Ana; Santos, Catarina; Almeida, Maria Margarida; Costa, Maria Elisabete; Fernandes, Maria Helena

    2014-12-01

    Nanosized hydroxyapatite (HA) is a promising material in clinical applications targeting the bone tissue. NanoHA is able to modulate bone cellular events, which accounts for its potential utility, but also raises safety concerns regarding the maintenance of the bone homeostasis. This work analyses the effects of HA nanoparticles (HAnp) on osteoclastic differentiation and activity, an issue that has been barely addressed. Rod-like HAnp, produced by a hydrothermal precipitation method, were tested on peripheral blood mononuclear cells (PBMC), which contains the CD14+ osteoclastic precursors, in unstimulated or osteoclastogenic-induced conditions. HAnp were added at three time-points during the osteoclastic differentiation pathway, and cell response was evaluated for osteoclastic related parameters. Results showed that HAnp modulated the differentiation and function of osteoclastic cells in a dose- and time-dependent manner. In addition, the effects were dependent on the stage of osteoclastic differentiation. In unstimulated PBMC, HAnp significantly increased osteoclastogenesis, leading to the formation of mature osteoclasts, as evident by the significant increase of TRAP activity, number of TRAP-positive multinucleated cells, osteoclastic gene expression and resorbing ability. However, in a population of mature osteoclasts (formed in osteoclastogenic-induced PBMC cultures), HAnp caused a dose-dependent decrease on the osteoclastic-related parameters. These results highlight the complex effects of HAnp in osteoclastic differentiation and activity, and suggest the possibility of HAnp to modulate/disrupt osteoclastic behavior, with eventual imbalances in the bone metabolism. This should be carefully considered in bone-related and other established and prospective biomedical applications of HAnp. PMID:26000372

  16. Pathogenesis of Age-Related Bone Loss in Humans

    PubMed Central

    2013-01-01

    Background. Although data from rodent systems are extremely useful in providing insights into possible mechanisms of age-related bone loss, concepts evolving from animal models need to ultimately be tested in humans. Methods. This review provides an update on mechanisms of age-related bone loss in humans based on the author’s knowledge of the field and focused literature reviews. Results. Novel imaging, experimental models, biomarkers, and analytic techniques applied directly to human studies are providing new insights into the patterns of bone mass acquisition and loss as well as the role of sex steroids, in particular estrogen, on bone metabolism and bone loss with aging in women and men. These studies have identified the onset of trabecular bone loss at multiple sites that begins in young adulthood and remains unexplained, at least based on current paradigms of the mechanisms of bone loss. In addition, estrogen appears to be a major regulator of bone metabolism not only in women but also in men. Studies assessing mechanisms of estrogen action on bone in humans have identified effects of estrogen on RANKL expression by several different cell types in the bone microenvironment, a role for TNF-α and IL-1β in mediating effects of estrogen deficiency on bone, and possible regulation of the Wnt inhibitor, sclerostin, by estrogen. Conclusions. There have been considerable advances in our understanding of age-related bone loss in humans. However, there are also significant gaps in knowledge, particularly in defining cell autonomous changes in bone in human studies to test or validate concepts emerging from studies in rodents. Decision Editor: Luigi Ferrucci, MD, PhD PMID:22923429

  17. Ex-vivo transduced autologous skin fibroblasts expressing human Lim Mineralization Protein-3 efficiently form new bone in animal models

    PubMed Central

    Lattanzi, Wanda; Parrilla, Claudio; Fetoni, Annarita; Logroscino, Giandomenico; Straface, Giuseppe; Pecorini, Giovanni; Stigliano, Egidio; Tampieri, Anna; Bedini, Rossella; Pecci, Raffaella; Michetti, Fabrizio; Gambotto, Andrea; Robbins, Paul D.; Pola, Enrico

    2012-01-01

    Local gene transfer of the human LIM Mineralization Protein (LMP), a novel intracellular positive regulator of the osteoblast differentiation program, can induce efficient bone formation in rodents. In order to develop a clinically relevant gene therapy approach to facilitate bone healing, we have used primary dermal fibroblasts transduced ex vivo with Ad.LMP3 and seeded on an hydroxyapatite/collagen matrix prior to autologous implantation. Here we demonstrate that genetically modified autologous dermal fibroblasts expressing Ad.LMP-3 are able to induce ectopic bone formation following implantation of the matrix into the mouse triceps and paravertebral muscles. Moreover, implantation of the Ad.LMP-3-modified dermal fibroblasts into a rat mandibular bone critical size defect model results in efficient healing as determined by X-ray, histology and three dimensional micro computed tomography (3DμCT). These results demonstrate the effectiveness of the non-secreted intracellular osteogenic factor LMP-3, in inducing bone formation in vivo. Moreover, the utilization of autologous dermal fibroblasts implanted on a biomaterial represents a promising approach for possible future clinical applications aimed at inducing new bone formation. PMID:18633445

  18. Bone scaffold architecture modulates the development of mineralized bone matrix by human embryonic stem cells

    PubMed Central

    Marcos-Campos, Ivan; Marolt, Darja; Petridis, Petros; Bhumiratana, Sarindr; Schmidt, Daniel; Vunjak-Novakovic, Gordana

    2012-01-01

    Decellularized bone has been widely used as a scaffold for bone formation, due to its similarity to the native bone matrix and excellent osteoinductive and biomechanical properties. We have previously shown that human mesenchymal and embryonic stem cells form functional bone matrix on such scaffolds, without the use of growth factors. In this study, we focused on differences in bone matrix that exist even among identical harvesting sites, and the effects of the matrix architecture and mineral content on bone formation by human embryonic stem cells (hESC). Mesenchymal progenitors derived from hESCs were cultured for 5 weeks in decellularized bone scaffolds with three different densities: low (0.281 ± 0.018 mg/mm3), medium (0.434 ± 0.015 mg/mm3) and high (0.618 ± 0.027 mg/mm3). The medium-density group yielded highest densities of cells and newly assembled bone matrix, presumably due to the best balance between the transport of nutrients and metabolites to and from the cells, space for cell infiltration, surface for cell attachment and the mechanical strength of the scaffolds, all of which depend on the scaffold density. Bone mineral was beneficial for the higher expression of bone markers in cultured cells and more robust accumulation of the new bone matrix. PMID:22901965

  19. Synergistic role of hydroxyapatite nanoparticles and pulsed electromagnetic field therapy to prevent bone loss in rats following exposure to simulated microgravity

    PubMed Central

    Prakash, D; Behari, J

    2009-01-01

    The purpose of the present study was to use capacitive coupling of pulsed electromagnetic field (CC-PEMF) and hydroxyapatite nanoparticles (HAp) as a countermeasure to prevent osteoporosis induced by simulated microgravity. We used the hind-limb suspension (HLS) rat model to simulate microgravity-induced bone losses for 45 days. In order to compare the resulting changes, mineralogical (bone mineral density [BMD], calcium [Ca], and phosphorus [P]), biochemical (osteocalcin, alkaline phosphatase [ALP], and type I collagen), and histological (scanning electron microscopy) parameters were adopted. As a countermeasure to the above, the effect of PEMF and HAp application were examined. Three-month-old female Wistar rats were randomly divided into control (n = 8), HLS (n = 8), HLS with PEMF (n = 8), HLS with HAp nanoparticles (n = 8), and HLS with HAp and PEMF (n = 8). We observed: 1) significant decrease (p < 0.01) in BMD, Ca, P, type I collagen, and ALP activity in femur and tibia in hind-limb bone and serum osteocalcin in HLS rats as compared with the ground control. 2) Nonsignificant increase in BMD (p < 0.1), Ca (p < 0.1), P (p < 0.5), type I collagen (p < 0.1), and ALP activity (p < 0.5) in femur and tibia in hind-limb bone and serum osteocalcin (p < 0.5) in HLS + PEMF rats compared with HLS rats. 3) Significant increase in BMD (p < 0.02), Ca (p < 0.05), P (p < 0.05), type I collagen (p < 0.02), and ALP activity (p > 0.02) in femur and tibia in hind-limb bone with a nonsignificant increase in serum osteocalcin (p > 0.1) in HLS + HAp rats compared to HLS rats. 4) Significant increase in BMD (p > 0.01). Ca (p > 0.01). P (p > 0.01). type I collagen (p > 0.01). and ALP activity (p > 0.01) in femur and tibia in hind-limb bone and serum osteocalcin (p > 0.02) were also observed. Results suggest that a combination of low level PEMF and Hap nanoparticles has potential to control bone loss induced by simulated microgravity. PMID:19774112

  20. Histological determination of the human origin of bone fragments.

    PubMed

    Cattaneo, Cristina; Porta, David; Gibelli, Daniele; Gamba, Corrado

    2009-05-01

    A frequently encountered task in the forensic scenario is verification of the human origin of severely degraded fragments of bone. In these cases histological methods which consider osteon size and morphology can prove to be useful. The authors in the present study verify the applicability of published algorithms to flat and subadult bones from human, dog, cat, cow, rabbit, sheep, pig, chicken, quail, and turkey samples. Metric analysis was performed on 2031 Haversian canals. Analyses carried out on human samples confirmed a success rate of around 70% on long adult bones; however the percentage of wrong answers was particularly high in the case of newborns and older subadults as well as on flat bones in general. Results therefore suggest that such regression equations should be limited only to bone fragments from long adult bones. PMID:19298464

  1. Incorporation of polymeric microparticles into collagen-hydroxyapatite scaffolds for the delivery of a pro-osteogenic peptide for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    López-Noriega, Adolfo; Quinlan, Elaine; Celikkin, Nehar; O'Brien, Fergal J.

    2015-01-01

    Collagen-hydroxyapatite scaffolds are outstanding materials for bone tissue engineering as they are biocompatible, bioresorbable, osteoconductive, and osteoinductive. The objective of the present work was to assess the potential of increasing their regenerative capacity by functionalising the scaffolds for therapeutic delivery. This was achieved by the utilization of polymeric drug carriers. With this purpose, alginate, chitosan, gelatine, and poly(lactic-co-glycolic acid) (PLGA) microparticles eluting PTHrP 107-111, an osteogenic pentapeptide, were fabricated and tested by incorporating them into the scaffolds. Among them, PLGA microparticles show the most promising characteristics for use as drug delivery devices. Following the incorporation of the microparticles, the scaffolds maintained their interconnected porous structure and the mechanical properties of the materials were not adversely affected. In addition, the microparticles released all their PTHrP 107-111 cargo. Most importantly, the delivered peptide proved to be bioactive and promoted enhanced osteogenesis as assessed by alkaline phosphatase production and osteocalcin and osteopontin gene expression when pre-osteoblastic cells were seeded on the scaffolds. While the focus was on bone repair, the release system described in this study can be used for the delivery of therapeutics for healing and regeneration of a variety of tissue types depending on the type of collagen scaffold chosen.

  2. Towards the Design of 3D Fiber-Deposited Poly(ε-caprolactone)/lron-Doped Hydroxyapatite Nanocomposite Magnetic Scaffolds for Bone Regeneration.

    PubMed

    De Santis, Roberta; Russo, Alessandro; Gloria, Antonio; D'Amora, Ugo; Russo, Teresa; Panseri, Silvia; Sandri, Monica; Tampieri, Anna; Marcacci, Maurilio; Dediu, Valentin A; Wilde, Colin J; Ambrosio, Luigi

    2015-07-01

    In the past few years, researchers have focused on the design and development of three-dimensional (3D) advanced scaffolds, which offer significant advantages in terms of cell performance. The introduction of magnetic features into scaffold technology could offer innovative opportunities to control cell populations within 3D microenvironments, with the potential to enhance their use in tissue regeneration or in cell-based analysis. In the present study, 3D fully biodegradable and magnetic nanocomposite scaffolds for bone tissue engineering, consisting of a poly(ε-caprolactone) (PCL) matrix reinforced with iron-doped hydroxyapatite (FeHA) nanoparticles, were designed and manufactured using a rapid prototyping technique. The performances of these novel 3D PCL/FeHA scaffolds were assessed through a combination of theoretical evaluation, experimental in vitro analyses and in vivo testing in a rabbit animal model. The results from mechanical com- pression tests were consistent with FEM simulations. The in vitro results showed that the cell growth in the magnetized scaffolds was 2.2-fold greater than that in non-magnetized ones. In vivo experiments further suggested that, after only 4 weeks, the PCL/FeHA scaffolds were completely filled with newly formed bone, proving a good level of histocompatibility. All of the results suggest that the introduction of magnetic features into biocompatible materials may confer significant advantages in terms of 3D cell assembly. PMID:26307846

  3. Human Placenta-Derived Adherent Cells Prevent Bone loss, Stimulate Bone formation, and Suppress Growth of Multiple Myeloma in Bone

    PubMed Central

    Li, Xin; Ling, Wen; Pennisi, Angela; Wang, Yuping; Khan, Sharmin; Heidaran, Mohammad; Pal, Ajai; Zhang, Xiaokui; He, Shuyang; Zeitlin, Andy; Abbot, Stewart; Faleck, Herbert; Hariri, Robert; Shaughnessy, John D.; van Rhee, Frits; Nair, Bijay; Barlogie, Bart; Epstein, Joshua; Yaccoby, Shmuel

    2011-01-01

    Human placenta has emerged as a valuable source of transplantable cells of mesenchymal and hematopoietic origin for multiple cytotherapeutic purposes, including enhanced engraftment of hematopoietic stem cells, modulation of inflammation, bone repair, and cancer. Placenta-derived adherent cells (PDACs) are mesenchymal-like stem cells isolated from postpartum human placenta. Multiple myeloma is closely associated with induction of bone disease and large lytic lesions, which are often not repaired and are usually the sites of relapses. We evaluated the antimyeloma therapeutic potential, in vivo survival, and trafficking of PDACs in the severe combined immunodeficiency (SCID)–rab model of medullary myeloma-associated bone loss. Intrabone injection of PDACs into non-myelomatous and myelomatous implanted bone in SCID-rab mice promoted bone formation by stimulating endogenous osteoblastogenesis, and most PDACs disappeared from bone within 4 weeks. PDACs inhibitory effects on myeloma bone disease and tumor growth were dose-dependent and comparable with those of fetal human mesenchymal stem cells (MSCs). Intrabone, but not subcutaneous, engraftment of PDACs inhibited bone disease and tumor growth in SCID-rab mice. Intratumor injection of PDACs had no effect on subcutaneous growth of myeloma cells. A small number of intravenously injected PDACs trafficked into myelomatous bone. Myeloma cell growth rate in vitro was lower in coculture with PDACs than with MSCs from human fetal bone or myeloma patients. PDACs also promoted apoptosis in osteoclast precursors and inhibited their differentiation. This study suggests that altering the bone marrow microenvironment with PDAC cytotherapy attenuates growth of myeloma and that PDAC cytotherapy is a promising therapeutic approach for myeloma osteolysis. PMID:21732484

  4. Fundamental ratios and logarithmic periodicity in human limb bones

    PubMed Central

    Pietak, Alexis; Ma, Siyan; Beck, Caroline W; Stringer, Mark D

    2013-01-01

    Fundamental mathematical relationships are widespread in biology yet there is little information on this topic with regard to human limb bone lengths and none related to human limb bone volumes. Forty-six sets of ipsilateral upper and lower limb long bones and third digit short bones were imaged by computed tomography. Maximum bone lengths were measured manually and individual bone volumes calculated from computed tomography images using a stereologic method. Length ratios of femur : tibia and humerus : ulna were remarkably similar (1.21 and 1.22, respectively) and varied little (<7%) between individuals. The volume ratio of femur : tibia was approximately half that of humerus : ulna (1.58 and 3.28, respectively; P < 0.0001). Lower limb bone volume ratios varied much more than upper limb ratios. The relationship between bone length and volume was found to be well described by power laws, with R2 values ranging from 0.983 to 0.995. The most striking finding was a logarithmic periodicity in bone length moving from distal to proximal up the limb (upper limb λ = 0.72, lower limb λ = 0.93). These novel data suggest that human limb bone lengths and volumes follow fundamental and highly conserved mathematical relationships, which may contribute to our understanding of normal and disordered growth, stature estimation, and biomechanics. PMID:23521756

  5. Facile green synthesis of silver doped fluor-hydroxyapatite/β-cyclodextrin nanocomposite in the dual acting fluorine-containing ionic liquid medium for bone substitute applications

    NASA Astrophysics Data System (ADS)

    Jegatheeswaran, S.; Selvam, S.; Sri Ramkumar, V.; Sundrarajan, M.

    2016-05-01

    A novel green route has approached for the synthesis of silver doped fluor-hydroxyapatite/β-cyclodextrin composite by the assistance of fluorine-based ionic liquid. The selected [BMIM]BF4 ionic liquid for this work plays a dual role as fluoride source and templating agent. It helps to improve the crystalline structures and the shape of the composites. The crystallinity, surface morphology, topographical studies of the synthesized composite were validated. The XRD results of the composite show typical Ag reflection peaks at 38.1°, 44.2° and 63.4°. The ionic liquid assisted composite displayed the hexagonal shaped HA particles, which are surrounded by spherical nano-Ag particles and these particles are uniformly dispersed in the β-cyclodextrin matrix in both horizontal and cross sections from surface morphology observations. The Ionic liquid assisted silver doped fluor-hydroxyapatite/β-cyclodextrin composite exhibited very good antibacterial activities against Escherichia coli, Salmonella typhi, Klebsiella pneumonia and Serratia liquefaciens pathogens. The antibacterial proficiencies were established using Confocal Laser Scanning Microscopic developed biofilms images and bacterial growth curve analysis. The cytotoxicity results of the ionic liquid assisted composite analyzed by cell proliferation in vitro studies using human osteosarcoma cell line (MG-63) and this study has shown excellent biocompatibility.

  6. Characterization of porous hydroxyapatite.

    PubMed

    Hing, K A; Best, S M; Bonfield, W

    1999-03-01

    Hydroxyapatite has been considered for use in the repair of osseous defects for the last 20 years. Recent developments have led to interest in the potential of porous hydroxyapatite as a synthetic bone graft. However, despite considerable activity in this field, regarding assessment of the biological response to such materials, the basic materials characterization is often inadequate. This paper documents the characterization of the chemical composition, mechanical integrity, macro- and microstructure of a porous hydroxyapatite, Endobon (E. Merck GmbH), intended for the bone-graft market. Specimens possesed a range of apparent densities from 0.35 to 1.44 g cm(-3). Chemical analysis demonstrated that the natural apatite precursor of Endobon was not converted to pure hydroxyapatite, but retained many of the ionic substituents found in bone mineral, notably carbonate, sodium and magnesium ions. Investigation of the microstructure illustrated that the struts of the material were not fully dense, but had retained some traces of the network of osteocyte lacunae. Macrostructural analysis demonstrated the complex inter-relationship between the structural features of an open pore structure. Both pore size and connectivity were found to be inversely dependent on apparent density. Furthermore, measurement of pore aspect ratio and orientation demonstrated a relationship between apparent density and the degree of macrostructural anisotropy within the specimens, while, it was also noted that pore connectivity was sensitive to anisotropy. Compression testing demonstrated the effect of apparent density and macrostructural anisotropy on the mechanical properties. An increase in apparent density from 0.38 to 1.25 g cm(-3) resulted in increases in ultimate compressive stress and compressive modulus of 1 to 11 MPa and 0.2 to 3.1 GPa, respectively. Furthermore, anisotropic high density (> 0.9 g cm(-3)) specimens were found to possess lower compressive moduli than isotropic specimens

  7. Ferroelectric polarization in nanocrystalline hydroxyapatite thin films on silicon.

    PubMed

    Lang, S B; Tofail, S A M; Kholkin, A L; Wojtaś, M; Gregor, M; Gandhi, A A; Wang, Y; Bauer, S; Krause, M; Plecenik, A

    2013-01-01

    Hydroxyapatite nanocrystals in natural form are a major component of bone--a known piezoelectric material. Synthetic hydroxyapatite is widely used in bone grafts and prosthetic pyroelectric coatings as it binds strongly with natural bone. Nanocrystalline synthetic hydroxyapatite films have recently been found to exhibit strong piezoelectricity and pyroelectricity. While a spontaneous polarization in hydroxyapatite has been predicted since 2005, the reversibility of this polarization (i.e. ferroelectricity) requires experimental evidence. Here we use piezoresponse force microscopy to demonstrate that nanocrystalline hydroxyapatite indeed exhibits ferroelectricity: a reversal of polarization under an electrical field. This finding will strengthen investigations on the role of electrical polarization in biomineralization and bone-density related diseases. As hydroxyapatite is one of the most common biocompatible materials, our findings will also stimulate systematic exploration of lead and rare-metal free ferroelectric devices for potential applications in areas as diverse as in vivo and ex vivo energy harvesting, biosensing and electronics. PMID:23884324

  8. Ferroelectric Polarization in Nanocrystalline Hydroxyapatite Thin Films on Silicon

    NASA Astrophysics Data System (ADS)

    Lang, S. B.; Tofail, S. A. M.; Kholkin, A. L.; Wojtaś, M.; Gregor, M.; Gandhi, A. A.; Wang, Y.; Bauer, S.; Krause, M.; Plecenik, A.

    2013-07-01

    Hydroxyapatite nanocrystals in natural form are a major component of bone- a known piezoelectric material. Synthetic hydroxyapatite is widely used in bone grafts and prosthetic pyroelectric coatings as it binds strongly with natural bone. Nanocrystalline synthetic hydroxyapatite films have recently been found to exhibit strong piezoelectricity and pyroelectricity. While a spontaneous polarization in hydroxyapatite has been predicted since 2005, the reversibility of this polarization (i.e. ferroelectricity) requires experimental evidence. Here we use piezoresponse force microscopy to demonstrate that nanocrystalline hydroxyapatite indeed exhibits ferroelectricity: a reversal of polarization under an electrical field. This finding will strengthen investigations on the role of electrical polarization in biomineralization and bone-density related diseases. As hydroxyapatite is one of the most common biocompatible materials, our findings will also stimulate systematic exploration of lead and rare-metal free ferroelectric devices for potential applications in areas as diverse as in vivo and ex vivo energy harvesting, biosensing and electronics.

  9. Ferroelectric Polarization in Nanocrystalline Hydroxyapatite Thin Films on Silicon

    PubMed Central

    Lang, S. B.; Tofail, S. A. M.; Kholkin, A. L.; Wojtaś, M.; Gregor, M.; Gandhi, A. A.; Wang, Y.; Bauer, S.; Krause, M.; Plecenik, A.

    2013-01-01

    Hydroxyapatite nanocrystals in natural form are a major component of bone- a known piezoelectric material. Synthetic hydroxyapatite is widely used in bone grafts and prosthetic pyroelectric coatings as it binds strongly with natural bone. Nanocrystalline synthetic hydroxyapatite films have recently been found to exhibit strong piezoelectricity and pyroelectricity. While a spontaneous polarization in hydroxyapatite has been predicted since 2005, the reversibility of this polarization (i.e. ferroelectricity) requires experimental evidence. Here we use piezoresponse force microscopy to demonstrate that nanocrystalline hydroxyapatite indeed exhibits ferroelectricity: a reversal of polarization under an electrical field. This finding will strengthen investigations on the role of electrical polarization in biomineralization and bone-density related diseases. As hydroxyapatite is one of the most common biocompatible materials, our findings will also stimulate systematic exploration of lead and rare-metal free ferroelectric devices for potential applications in areas as diverse as in vivo and ex vivo energy harvesting, biosensing and electronics. PMID:23884324

  10. Electrospun poly (3-hydroxybutyrate-co-3-hydroxyvalerate)/hydroxyapatite scaffold with unrestricted somatic stem cells for bone regeneration.

    PubMed

    Biazar, Esmaeil; Heidari Keshel, Saeed

    2015-01-01

    The combination of scaffolds and cells can be useful in tissue reconstruction. In this study, nanofibrous poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)/nanohydroxyapatite (nano-HAp) scaffolds, filled with unrestricted somatic stem cells (USSCs), were used for healing calvarial bone in rat model. The healing effects of these scaffolds, with and without stem cells, in bone regeneration were investigated by computed tomography (CT) analysis and pathology assays after 28 days of grafting. The results of CT analysis showed that bone regeneration on the scaffolds, and the amounts of regenerated new bone for polymer/nano-HAp scaffold with USSC, was significantly greater than the scaffold without cell and untreated control samples. Therefore, the combination of scaffold especially with USSC could be considered as a useful method for bone regeneration. PMID:25710767

  11. Early Osteogenic Signal Expression of Rat Bone Marrow Stromal Cells is Influenced by Both Hydroxyapatite Nanoparticle Content and Initial Cell Seeding Density in Biodegradable Nanocomposite Scaffolds

    PubMed Central

    Kim, Kyobum; Dean, David; Lu, Anqi; Mikos, Antonios G.; Fisher, John P.

    2010-01-01

    Incorporation of hydroxyapatite (HA) within a degradable polymeric scaffold may provide a favorable synthetic microenvironment that more closely mimics natural bone tissue physiology. Both incorporation of HA nanoparticles and alteration of paracrine cell-cell signaling distances may affect the intercellular signaling mechanism and facilitate the enhanced osteogenic signal expressions among the implanted cell population. In this study, we investigate the effect of the incorporation of HA nanoparticles into poly(propylene fumarate) (PPF) scaffolds on the surface properties of composite scaffolds and early osteogenic growth factor gene expression in relation to initial cell seeding density. The result of surface characterization indicated that HA addition improved surface properties of PPF/HA composite scaffolds by showing increased roughness, hydrophilicity, protein adsorption, and initial cell attachment. Rat bone marrow stromal cells (BMSCs), which were CD34(−), CD45(−), CD29(+), and CD90(+), were cultured on 3D macroporous PPF/HA scaffolds with two different initial cell seeding densities (0.33 and 1.00 million cells per scaffold) for 8 days. Results demonstrated that endogenous osteogenic signal expression profiles, including bone morphogenetic protein-2, fibroblast growth factor-2, and transforming growth factor-β1, as well as the transcriptional factor Runx2 were affected by both HA amount and initial cell seeding density. Upregulated expression of osteogenic growth factor genes was related to subsequent osteoblastic differentiation of rat BMSCs on 3D scaffolds, as characterized by alkaline phosphatase activity, osteocalcin mRNA expression, and calcium deposition. Thus PPF/HA composite scaffold construction parameters, including incorporated HA amount and initial cell seeding density, may be utilized to induce the osteoblastic differentiation of transplanted rat BMSCs. PMID:21074640

  12. Nano-crystalline hydroxyapatite bone graft combined with bioresorbable collagen membrane in the treatment of periodontal intrabony defects: A randomized controlled clinical trial

    PubMed Central

    Singh, Vijendra P.; Nayak, Dilip G.; Uppoor, Ashita S.; Shah, Dipen

    2012-01-01

    Aim: To evaluate the clinical outcome of nanocrystalline hydroxyapatite (NcHA) bonegraft (Sybograf®) in combination with collagen membrane (PerioCol®) compared with open flap debridement (OFD) only in the treatment of intrabony periodontal defects. Materials and Methods: Eighteen intrabony defects in 16 systemically healthy patients aged between 25-65 years, were randomly assigned to test and control groups. The Plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL), and gingival recession were recorded at baseline, and were reevaluated at 6 months. In addition to this, radiographic bone fill was assessed using digital software. At the test site NcHA bone graft and collagen membrane was placed, whereas at the control site only, OFD was done. Recall appointments were made at 7th day, 1st month, 3rd month, and 6th month. Results: The data were subjected to statistical analysis using the Mann-Whitney ‘U’ Test and Wilcoxon signed rank sum test. In the control group, the mean reduction of PPD was 3.22±1.09 mm and CAL gain was 2.78±1.09 mm. In the test group, the mean PPD reduction of 4.33±0.5 mm and mean gain in CAL was 3.78±0.66 mm at 6 months. The mean increase in gingival recession was 0.55±0.72 mm in test and 0.44±0.52 mm in control group. Conclusion: The NcHA bone graft in combination with collagen membrane demonstrated better clinical outcomes compared with OFD alone. PMID:23493628

  13. Treatment of Staphylococcus aureus-induced chronic osteomyelitis with bone-like hydroxyapatite/poly amino acid loaded with rifapentine microspheres

    PubMed Central

    Yan, Ling; Jiang, Dian-Ming; Cao, Zhi-Dong; Wu, Jun; Wang, Xin; Wang, Zheng-Long; Li, Ya-Jun; Yi, Yong-Fen

    2015-01-01

    Purpose The purpose of this study was to investigate the curative effect of bone-like hydroxyapatite/poly amino acid (BHA/PAA) as a carrier for poly(lactic-co-glycolic acid)-coated rifapentine microsphere (RPM) in the treatment of rabbit chronic osteomyelitis induced by Staphylococcus aureus. Methods RPM was prepared through an oil-in-water emulsion solvent evaporation method, and RPM was combined with BHA/PAA to obtain drug-loaded, slow-releasing materials. Twenty-six New Zealand white rabbits were induced to establish the animal model of chronic osteomyelitis. After debridement, the animals were randomly divided into three groups (n=8): the experimental group (with RPM-loaded BHA/PAA), the control group (with BHA/PAA), and the blank group. The RPM-loaded BHA/PAA was evaluated for antibacterial activity, dynamics of drug release, and osteogenic ability through in vitro and in vivo experiments. Results In vitro, RPM-loaded BHA/PAA released the antibiotics slowly, inhibiting the bacterial growth of S. aureus for up to 5 weeks. In vivo, at week 4, the bacterial colony count was significantly lower in the experimental group than in the control and blank groups (P<0.01). At week 12, the chronic osteomyelitis was cured and the bone defect was repaired in the experimental group, whereas the infection and bone defect persisted in the control and blank groups. Conclusion In vitro and in vivo experiments demonstrated that RPM-loaded BHA/PAA effectively cured S. aureus-induced chronic osteomyelitis. Therefore, BHA/PAA has potential value as a slow-releasing material in clinical setting. Further investigation is needed to determine the optimal dosage for loading rifapentine. PMID:26213463

  14. Property of Human Bone Marrow Stromal Cells Derived From Bone Fragments Removed in Sagittal Split Ramus Osteotomy.

    PubMed

    Yoshida, Chihiro; Yamaguchi, Satoshi; Abe, Shigehiro; Harada, Kiyoshi

    2016-06-01

    Bone tissue engineering is in the process of making the shift from bench to bed. Organ as a cell source is important for tissue engineering. The appropriate cells should be harvested without invasiveness and ethical problems. The authors focused on mandibular cortex bone fragments removed in sagittal split ramus osteotomy as a cell source for bone tissue engineering. These bone fragments were discarded after surgery until now. Bone marrow stromal cells (BMSCs) were harvested from inside of bone fragments, which is an endosteal region. Endosteal region is known to be a hematopoietic stem cell niche and harbors osteoblasts, preosteoblasts, and mesenchymal stem cells (MSCs). Bone marrow stromal cells could be cultured easily, and grew rapidly in vitro under ordinary serum-supplemented culture condition. The expression pattern of surface markers of BMSCs was the same as that of MSCs. Bone marrow stromal cells could differentiated into multiple mesenchymal lineages (osteoblasts, adipocytes, chondrocytes, and smooth muscle cells). These results indicated the existence of MSCs in BMSCs. The osteoblastic characters of BMSCs were examined more closely. Bone marrow stromal cells showed a high alkaline phosphatase activity, and expressed osteoblastic markers (PTHr, bone sialoprotein, Type I collagen, Rnut-related transcription factor 2, and osteocalcin). In transplantation experiments, BMSCs generated ectopic bone tissues on the border of hydroxyapatite scaffold without osteogenic differentiation-inducing agents such as dexamethasone (Dex) or bone morphogenetic protein. The results of this study suggest that mandibular cortex bone fragments removed in sagittal split ramus osteotomy are a good cell source for bone tissue engineering. PMID:27171960

  15. Scanning acoustic microscopy study of human cortical and trabecular bone.

    PubMed

    Bumrerraj, S; Katz, J L

    2001-12-01

    Scanning acoustic microscopy (SAM) has been used in the burst mode to study the properties of human cortical and trabecular bone. An Olympus UH3 SAM (Olympus Co., Tokyo, Japan) was used with a 400 MHz burst mode lens (120 degrees aperture, nominal lateral resolution 2.5 microm). The human cortical bone was from the midshaft of a femur from a 60+ male cadaver; the trabecular bone specimens were obtained from the distal femoral condyles of another 60+ human male cadaver. Elastic moduli for both trabecular and cortical bone were obtained by means of a series of calibration curves correlating SAM gray levels of known materials with their elastic moduli; specimens included: polypropylene, PMMA, Teflon, aluminum, Pyrex glass, titanium, and stainless steel. Values obtained by this method are in good agreement with those obtained by nanoindentation techniques. The three critical findings earlier by Katz and Meunier were observed here as well in both the cortical and trabecular bone samples. PMID:11853252

  16. Comparative cortical bone thickness between the long bones of humans and five common non-human mammal taxa.

    PubMed

    Croker, Sarah L; Reed, Warren; Donlon, Denise

    2016-03-01

    The task of identifying fragments of long bone shafts as human or non-human is difficult but necessary, for both forensic and archaeological cases, and a fast simple method is particularly useful. Previous literature suggests there may be differences in the thickness of the cortical bone between these two groups, but this has not been tested thoroughly. The aim of this study was not only to test this suggestion, but also to provide data that could be of practical assistance for future comparisons. The major limb bones (humerus, radius, femur and tibia) of 50 Caucasoid adult skeletons of known age and sex were radiographed, along with corresponding skeletal elements from sheep, pigs, cattle, large dogs and kangaroos. Measurements were taken from the radiographs at five points along the bone shaft, of shaft diameter, cortical bone thickness, and a cortical thickness index (sum of cortices divided by shaft diameter) in both anteroposterior and mediolateral orientations. Each variable for actual cortical bone thickness as well as cortical thickness indices were compared between the human group (split by sex) and each of the non-human groups in turn, using Student's t-tests. Results showed that while significant differences did exist between the human groups and many of the non-human groups, these were not all in the same direction. That is, some variables in the human groups were significantly greater than, and others were significantly less than, the corresponding variable in the non-human groups, depending on the particular non-human group, sex of the human group, or variable under comparison. This was the case for measurements of both actual cortical bone thickness and cortical thickness index. Therefore, for bone shaft fragments for which the skeletal element is unknown, the overlap in cortical bone thickness between different areas of different bones is too great to allow identification using this method alone. However, by providing extensive cortical bone

  17. Revision stapes surgery for lysis of the long process of the incus: comparing hydroxyapatite bone cement versus malleovestibulopexy and total ossicular replacement prosthesis.

    PubMed

    Pitiot, Vincent; Hermann, Ruben; Tringali, Stéphane; Dubreuil, Christian; Truy, Eric

    2016-09-01

    The objective of the study was to report audiological results in revision stapes surgery, comparing hydroxyapatite (HAP) bone cement, malleovestibular (MV) prosthesis, and total ossicular replacement prosthesis (TORP). The study is a retrospective case review conducted in a tertiary referral center. Patients treated for revision stapes surgery from 2010 to 2014, where a lysis of the long process of the incus (LPI) was observed with the use of HAP bone cement, MV prosthesis, or a TORP were included in the study. The main outcomes measured were pre- and postoperative bone conduction (BC) and air conduction (AC) pure-tone averages (PTA) (0.5, 1, 2, 3 kHz), including high frequencies BC (HFBC) (1, 2, 3, 4 kHz) and air-bone gap (ABG). 107 revision stapes surgery were performed in 96 ears. Main cause of failure was LPI lysis in 38 cases (39.6 %). 31 patients were analyzed: HAP bone cement was used in 11 patients (Group I), MV prosthesis in ten patients (Group II), and TORP in ten patients (Group III). The mean post-operative ABG was 10.7 dB (±7.4) (p = 0.003), 10.7 dB (±8.8) (p = 0.001), and 16.9 dB (±9.8) (p = 0.001), respectively. There were no significant differences between groups. In Group I, the mean change in HFBC revealed an improvement of 5.6 dB (±7.9) (p = 0.03), while in Group III there was a significant deterioration of the thresholds of 5.8 dB (±7.6) (p = 0.04). There were no cases of post-operative anacusis. In revision stapes surgery when LPI is eroded, we recommend to perform a cement ossiculoplasty for stabilizing a standard Teflon piston when LPI is still usable, the LPI lengthening with cement being not recommended. When LPI is too eroded, we prefer performing a malleovestibulopexy, and reserve TORP for cases with a bad anatomical presentation. PMID:26690574

  18. Cross-correlative 3D micro-structural investigation of human bone processed into bone allografts.

    PubMed

    Singh, Atul Kumar; Gajiwala, Astrid Lobo; Rai, Ratan Kumar; Khan, Mohd Parvez; Singh, Chandan; Barbhuyan, Tarun; Vijayalakshmi, S; Chattopadhyay, Naibedya; Sinha, Neeraj; Kumar, Ashutosh; Bellare, Jayesh R

    2016-05-01

    Bone allografts (BA) are a cost-effective and sustainable alternative in orthopedic practice as they provide a permanent solution for preserving skeletal architecture and function. Such BA however, must be processed to be disease free and immunologically safe as well as biologically and clinically useful. Here, we have demonstrated a processing protocol for bone allografts and investigated the micro-structural properties of bone collected from osteoporotic and normal human donor samples. In order to characterize BA at different microscopic levels, a combination of techniques such as Solid State Nuclear Magnetic Resonance (ssNMR), Scanning Electron Microscope (SEM), micro-computed tomography (μCT) and Thermal Gravimetric Analysis (TGA) were used for delineating the ultra-structural property of bone. ssNMR revealed the extent of water, collagen fine structure and crystalline order in the bone. These were greatly perturbed in the bone taken from osteoporotic bone donor. Among the processing methods analyzed, pasteurization at 60 °C and radiation treatment appeared to substantially alter the bone integrity. SEM study showed a reduction in Ca/P ratio and non-uniform distribution of elements in osteoporotic bones. μ-CT and MIMICS (Materialize Interactive Medical Image Control System) demonstrated that pasteurization and radiation treatment affects the BA morphology and cause a shift in the HU unit. However, the combination of all these processes restored all-important parameters that are critical for BA integrity and sustainability. Cross-correlation between the various probes we used quantitatively demonstrated differences in morphological and micro-structural properties between BA taken from normal and osteoporotic human donor. Such details could also be instrumental in designing an appropriate bone scaffold. For the best restoration of bone microstructure and to be used as a biomaterial allograft, a step-wise processing method is recommended that preserves all

  19. Osteogenic differentiation of human bone marrow mesenchymal stem cells in hydrogel containing nacre powder.

    PubMed

    Flausse, Alicia; Henrionnet, Christel; Dossot, Manuel; Dumas, Dominique; Hupont, Sébastien; Pinzano, Astrid; Mainard, Didier; Galois, Laurent; Magdalou, Jacques; Lopez, Evelyne; Gillet, Pierre; Rousseau, Marthe

    2013-11-01

    Nacre (or mother of pearl) can facilitate bone cell differentiation and can speed up their mineralization. Here we report on the capability of nacre to induce differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) and the production of extracellular matrix. hBM-MSCs were encapsulated in an alginate hydrogel containing different concentrations of powdered nacre and cultured in the same environment until Day 28. Analysis of osteogenic gene expression, histochemistry, second harmonic generation (SHG) microscopy, and Raman scattering spectroscopy were used to characterize the synthesis of the extracellular matrix. In the presence of nacre powder, a significant increase in matrix synthesis from D21 in comparison with pure alginate was observed. Histochemistry revealed the formation of a new tissue composed of collagen fibers in the presence of nacre (immunostaining and SHG), and hydroxyapatite crystals (Raman) in the alginate beads. These results suggest that nacre is efficient in hBM-MSCs differentiation, extracellular matrix production and mineralization in alginate 3D biomaterials. PMID:23554327

  20. Hydroxyapatite-doped polycaprolactone nanofiber membrane improves tendon-bone interface healing for anterior cruciate ligament reconstruction.

    PubMed

    Han, Fei; Zhang, Peng; Sun, Yaying; Lin, Chao; Zhao, Peng; Chen, Jiwu

    2015-01-01

    Hamstring tendon autograft is a routine graft for anterior cruciate ligament (ACL) reconstruction. However, ways of improving the healing between the tendon and bone is often overlooked in clinical practice. This issue can be addressed by using a biomimetic scaffold. Herein, a biomimetic nanofiber membrane of polycaprolactone/nanohydroxyapatite/collagen (PCL/nHAp/Col) is fabricated that mimics the composition of native bone tissue for promoting tendon-bone healing. This membrane has good cytocompatibility, allowing for osteoblast cell adhesion and growth and bone formation. As a result, MC3T3 cells reveal a higher mineralization level in PCL/nHAp/Col membrane compared with PCL membrane alone. Further in vivo studies in ACL reconstruction in a rabbit model shows that PCL/nHAp/Col-wrapped tendon may afford superior tissue integration to nonwrapped tendon in the interface between the tendon and host bone as well as improved mechanical strength. This study shows that PCL/nHAp/Col nanofiber membrane wrapping of autologous tendon is effective for improving tendon healing with host bone in ACL reconstruction. PMID:26677323

  1. Hydroxyapatite-doped polycaprolactone nanofiber membrane improves tendon–bone interface healing for anterior cruciate ligament reconstruction

    PubMed Central

    Han, Fei; Zhang, Peng; Sun, Yaying; Lin, Chao; Zhao, Peng; Chen, Jiwu

    2015-01-01

    Hamstring tendon autograft is a routine graft for anterior cruciate ligament (ACL) reconstruction. However, ways of improving the healing between the tendon and bone is often overlooked in clinical practice. This issue can be addressed by using a biomimetic scaffold. Herein, a biomimetic nanofiber membrane of polycaprolactone/nanohydroxyapatite/collagen (PCL/nHAp/Col) is fabricated that mimics the composition of native bone tissue for promoting tendon–bone healing. This membrane has good cytocompatibility, allowing for osteoblast cell adhesion and growth and bone formation. As a result, MC3T3 cells reveal a higher mineralization level in PCL/nHAp/Col membrane compared with PCL membrane alone. Further in vivo studies in ACL reconstruction in a rabbit model shows that PCL/nHAp/Col-wrapped tendon may afford superior tissue integration to nonwrapped tendon in the interface between the tendon and host bone as well as improved mechanical strength. This study shows that PCL/nHAp/Col nanofiber membrane wrapping of autologous tendon is effective for improving tendon healing with host bone in ACL reconstruction. PMID:26677323

  2. Bioreactor Cultivation of Anatomically Shaped Human Bone Grafts

    PubMed Central

    Temple, Joshua P.; Yeager, Keith; Bhumiratana, Sarindr; Vunjak-Novakovic, Gordana; Grayson, Warren L.

    2015-01-01

    In this chapter, we describe a method for engineering bone grafts in vitro with the specific geometry of the temporomandibular joint (TMJ) condyle. The anatomical geometry of the bone grafts was segmented from computed tomography (CT) scans, converted to G-code, and used to machine decellularized trabecular bone scaffolds into the identical shape of the condyle. These scaffolds were seeded with human bone marrow-derived mesenchymal stem cells (MSCs) using spinner flasks and cultivated for up to 5 weeks in vitro using a custom-designed perfusion bioreactor system. The flow patterns through the complex geometry were modeled using the FloWorks module of SolidWorks to optimize bioreactor design. The perfused scaffolds exhibited significantly higher cellular content, better matrix production, and increased bone mineral deposition relative to non-perfused (static) controls after 5 weeks of in vitro cultivation. This technology is broadly applicable for creating patient-specific bone grafts of varying shapes and sizes. PMID:24014312

  3. Bone mineral density, Bone mineral contents, MMP-8 and MMP-9 levels in Human Mandible and alveolar bone: Simulated microgravity

    NASA Astrophysics Data System (ADS)

    Rai, Balwant; Kaur, Jasdeep; Catalina, Maria

    Exposure to microgravity has been associated with several physiological changes in astronauts and cosmonauts, including an osteoporosis-like loss of bone mass. It has been reported that head-down tilt bed-rest studies mimic many of the observations seen in flights. There is no study on the correlation on effects of mandibular bone and alveolar bone loss in both sex in simulating microgravity. This study was designed to determine the Bone mineral density and GCF MMP-8 MMP-9 in normal healthy subject of both sexes in simulated microgravity condition of -6 head-down-tilt (HDT) bed rest. The subjects of this investigation were 10 male and 10 female volunteers participated in three weeks 6 HDT bed-rest exposure. The Bone density and bone mineral contents were measured by dual energy X-ray absorptiometry before and in simulated microgravity. The GCF MMP-8 MMP-8 were measured by Enzyme-linked immunosorbent assays (Human Quantikine MMP-8,-9 ELISA kit). The bone mineral density and bone mineral contents levels were significantly decreased in simulated microgravity condition in both genders, although insignificantly loss was higher in females as compared to males. MMP-8 MMP-9 levels were significantly increased in simulated microgravity as compared to normal condition although insignificantly higher in females as compared to males. Further study is required on large samples size including all factors effecting in simulated microgravity and microgravity. Keys words-Simulated microgravity condition, head-down-tilt, Bone loss, MMP-8, MMP-9, Bone density, Bone mineral contents.

  4. Degradation pattern of porous CaCO3 and hydroxyapatite microspheres in vitro and in vivo for potential application in bone tissue engineering.

    PubMed

    Zhong, Qiwei; Li, Wenhua; Su, Xiuping; Li, Geng; Zhou, Ying; Kundu, Subhas C; Yao, Juming; Cai, Yurong

    2016-07-01

    Despite superior clinical handling, excellent biocompatibility, biodegradation property of calcium phosphate needs to be improved to coincide with the rate of new bone formation. In this study, spherical CaCO3 are fabricated in the presence of the silk sericin and then transformed into porous hydroxyapatite (HAP) microspheres via hydrothermal method. The degradation behavior of obtained CaCO3, HAP and their mixture is first investigated in vitro. The result demonstrates that the weight loss of HAP microspheres are almost 24.3% after immersing in pH 7.40 Tris-HCl buffer solution for 12 weeks, which is far slower than that of spherical CaCO3 (97.5%). The degradation speed of the mixtures depends on the proportion of CaCO3 and HAP. The mixture with higher content of CaCO3 possesses a quicker degradation speed. The obtained CaCO3 and HAP microspheres are injected into subcutaneous tissue of ICR mice with the assistance of sodium alginate. The result in vivo also shows an obvious difference of degradation speed between the obtained CaCO3 and HAP microspheres, implying it is feasible to modulate the degradation property of the mixture through changing the proportion of CaCO3 and HAP The good cytocompatibility of the two kinds of microspheres is proved and a mild inflammation response is observed only at early stage of implantation. The job offers a simple method to modify the degradation properties of biomaterial for potential use in bone tissue engineering. PMID:26998866

  5. Mechanical testing of recombinant human bone morphogenetic protein-7 regenerated bone in sheep mandibles.

    PubMed

    Kontaxis, A; Abu-Serriah, M; Ayoub, A F; Barbenel, J C

    2004-01-01

    A new method was developed in this study for testing excised sheep mandibles as a cantilever. The method was used to determine the strength and stiffness of sheep hemi-mandibles including a 35 mm defect bridged by regenerated bone. Recombinant human bone morphogenetic protein-7 (rhBMP-7) in a bovine collagen type-I carrier was used for the bone regeneration. Initial tests on ten intact sheep mandibles confirmed that the strength, stiffness and area beneath the load-deformation curves of the right and left hemi-mandibles were not significantly different, confirming the validity of using the contra-lateral hemi-mandible as a control side. Complete bone regeneration occurred in six hemi-mandibles treated with rhBMP, but the quality and mechanical properties of the bone were very variable. The new bone in three samples contained fibrous tissue and was weaker and less stiff than the contra-lateral side (strength, 10-20 per cent; stiffness, 6-15 per cent). The other half had better-quality bone and was significantly stiffer and stronger (p < 0.05), with strength 45-63 per cent and stiffness 35-46 per cent of the contra-lateral side. Hemi-mandibles treated with collagen alone had no regenerated bone bridge suggesting that 35 mm is a critical-size bone defect. PMID:15648662

  6. Sequential extracts of human bone show differing collagen synthetic rates.

    PubMed

    Babraj, J; Cuthbertson, D J; Rickhuss, P; Meier-Augenstein, W; Smith, K; Bohé, J; Wolfe, R R; Gibson, J N A; Adams, C; Rennie, M J

    2002-04-01

    Type I collagen is the major bone protein. Little is known quantitatively about human bone collagen synthesis in vivo, despite its importance for the understanding of bone formation and turnover. Our aim was to develop a method that could be used for the physiological and pathophysiological investigation of human bone collagen synthesis. We have carried out preliminary studies in patients undergoing hip replacement and in pigs to validate the use of the flooding dose method using (13)C- or (15)N-labelled proline and we have now refined our techniques to allow them to be used in a normal clinical or physiological setting. The results show that the application of a flooding dose causes bone free-proline labelling to equilibrate with that of blood in pigs and human beings, so that only 150 mg of bone will provide enough sample to prepare and measure the labelling of three fractions of bone collagen (dissolved in NaCl, acetic acid and pepsin/acetic acid) which have the same relative labelling (1.0:0.43:0.1) as measured by GC-combustion-isotope ratio MS. The rates of incorporation were substantially faster than in skeletal muscle samples taken at the same time. The results suggest that different fractions of human bone collagen turnover at markedly higher rates than had been previously considered. This approach should allow us to discover how growth and development, food, activity and drugs affect bone collagen turnover and to measure the effects on it of ageing and bone disease. PMID:12023825

  7. Bone marrow stromal cells cultured on poly (lactide-co-glycolide)/nano-hydroxyapatite composites with chemical immobilization of Arg-Gly-Asp peptide and preliminary bone regeneration of mandibular defect thereof.

    PubMed

    Huang, Yanxia; Ren, Jie; Ren, Tianbin; Gu, Shuying; Tan, Qinggang; Zhang, Lihong; Lv, Kaige; Pan, Kefeng; Jiang, Xinquan

    2010-12-15

    Polyethyleneimine (PEI) was used to create active groups on the poly (lactide-co-glycolide)/nano-hydroxyapatite (PLGA/NHA) surface and Arg-Gly-Asp (RGD) was grafted on the active groups and novel PLGA/NHA 2-D membranes and 3D scaffolds modified with RGD were obtained. X-ray photoelectron spectrum (XPS) results show that sulfur displays only on the modified surface. The RGD-modified PLGA/NHA materials also have much lower static water contact angle and much higher water-absorption ability, which shows that after chemical treatment, the modified materials show better hydrophilic properties. Atomic force microscope (AFM) shows that after surface modification, the surface morphology of PLGA is greatly changed. All these results indicate that RGD peptide has successfully grafted on the surface of PLGA. Rabbit bone marrow stromal cells (MSCs) were seeded in the 2D membranes and 3D scaffolds materials. The influences of the RGD on the cell attachment, growth and differentiation, and proliferation on the different materials were studied. The modified scaffolds were implanted into rabbits to observe preliminary application in regeneration of mandibular defect. The PLGA/NHA-RGD presents better results in bone regeneration in rabbit mandibular defect. PMID:20872750

  8. Preparation of laminated poly(ε-caprolactone)-gelatin-hydroxyapatite nanocomposite scaffold bioengineered via compound techniques for bone substitution.

    PubMed

    Hamlekhan, Azhang; Moztarzadeh, Fathollah; Mozafari, Masoud; Azami, Mahmoud; Nezafati, Nader

    2011-01-01

    In this research, new bioactive nanocomposite scaffolds were successfully developed using poly(ε-caprolactone) (PCL), cross-linked gelatin and nanoparticles of hydroxyapatite (HAp) after testing different solvents and methods. First, HAp powder was synthesized via a chemical precipitation technique and characterized. Then, the nanocomposites were prepared through layer solvent casting combined with freeze-drying and lamination techniques. According to the results, the increasing of the PCL weight in the scaffolds led to the improvement of the mechanical properties. The amount of ultimate stress, stiffness and also elastic modulus increased from 8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL. The biomineralization study revealed the formation of an apatite layer on the scaffolds after immersion in simulated body fluid (SBF). The Ca-P ratios were in accordance to nonstoichiometric biological apatite, which was approximately 1.67. The in vitro biocompatibility and cytocompatibility of the scaffolds were tested using mesenchymal stem cells (MSCs), and the results indicated no sign of toxicity, and cells were found to be attached to the scaffold walls. The in vivo biocompatibility and osteogenesis of these scaffolds in the animal experiments is also under investigation, and the result will be published at the end of the study. PMID:23507731

  9. Preparation and characterization of nano-sized hydroxyapatite/alginate/chitosan composite scaffolds for bone tissue engineering.

    PubMed

    Kim, Hye-Lee; Jung, Gil-Yong; Yoon, Jun-Ho; Han, Jung-Suk; Park, Yoon-Jeong; Kim, Do-Gyoon; Zhang, Miqin; Kim, Dae-Joon

    2015-09-01

    The aim of this study was to develop chitosan composite scaffolds with high strength and controlled pore structures by homogenously dispersed nano-sized hydroxyapatite (nano-HAp) powders. In the fabrication of composite scaffolds, nano-HAp powders distributed in an alginate (AG) solution with a pH higher than 10 were mixed with a chitosan (CS) solution and then freeze dried. While the HAp content increased up to 70 wt.%, the compressive strength and the elastic modulus of the composite scaffolds significantly increased from 0.27 MPa and 4.42 MPa to 0.68 MPa and 13.35 MPa, respectively. Higher content of the HAp also helped develop more differentiation and mineralization of the MC3T3-E1 cells on the composite scaffolds. The uniform pore structure and the excellent mechanical properties of the HAp/CS composite scaffolds likely resulted from the use of the AG solution at pH 10 as a dispersant for the nano-HAp powders. PMID:26046263

  10. Generation of clinical grade human bone marrow stromal cells for use in bone regeneration.

    PubMed

    Robey, Pamela G; Kuznetsov, Sergei A; Ren, Jiaqiang; Klein, Harvey G; Sabatino, Marianna; Stroncek, David F

    2015-01-01

    In current orthopaedic practice, there is a need to increase the ability to reconstruct large segments of bone lost due to trauma, resection of tumors and skeletal deformities, or when normal regenerative processes have failed such as in non-unions and avascular necrosis. Bone marrow stromal cells (BMSCs, also known as bone marrow-derived mesenchymal stem cells), when used in conjunction with appropriate carriers, represent a means by which to achieve bone regeneration in such cases. While much has been done at the bench and in pre-clinical studies, moving towards clinical application requires the generation of clinical grade cells. What is described herein is an FDA-approved cell manufacturing procedure for the ex vivo expansion of high quality, biologically active human BMSCs. This article is part of a Special Issue entitled Stem Cells and Bone. PMID:25064527

  11. Estrogen enhances the bone regeneration potential of periodontal ligament stem cells derived from osteoporotic rats and seeded on nano-hydroxyapatite/collagen/poly(L-lactide)

    PubMed Central

    E, LING-LING; XU, WEN-HUAN; FENG, LIN; LIU, YI; CAI, DONG-QING; WEN, NING; ZHENG, WEN-JIE

    2016-01-01

    This study investigated the effects of estrogen on the bone regeneration potential of periodontal ligament stem cells (PDLSCs) derived from osteoporotic rats and seeded on a collagen-based composite scaffold [nano-hydroxyapatite/collagen/poly(L-lactide) (nHAC/PLA)]. For this purpose, 48 healthy 3-month-old Sprague-Dawley female rats were divided into 2 groups as follows: the bilaterally ovariectomized (OVX) rats and sham-operated rats. The PDLSCs were isolated at 3 months after surgery (by which time postmenopausal osteoporosis had developed). The effects of estrogen on the characteristics of these cells seeded in a culture plate and of the cells seeded on nHAC/PLA were then investigated. The PDLSC + nHAC/PLA constructs were implanted subcutaneously into the backs of severe combined immunodeficient (SCID) mice for 12 weeks in order to examine the role of estrogen in the bone formation ability of PDLSCs derived from osteoporotic rats. The results from methyl thiazolyl tetrazolium (MTT) assay revealed that the proliferation of the cells derived from the rats in the OVX group was significantly higher than that of the cells derived from the rats in the sham-operated group at the stage of logarithmic growth. The staining intensity of alkaline phosphatase (ALP) and the mineralization of the cells derived from the rats in the OVX group was significantly weaker than that of the cells from the rats in the sham-operated group. When the PDLSCs were seeded on nHAC/PLA, ALP activity, osteocalcin (OCN) secretion, mineral formation and the mRNA expression levels of ALP, OCN, estrogen receptor (ER)α and ERβ in the cells derived from the rats in the OVX group were markedly decreased. Treatment with 17β-estradiol (E2) significantly weakened the proliferative ability of the cells derived from the OVX group rats, and enhanced their osteogenic differentiation ability and the mRNA expression levels of ALP, OCN, ERα and ERβ. When the constructs were implanted into the backs of SCID

  12. Estrogen enhances the bone regeneration potential of periodontal ligament stem cells derived from osteoporotic rats and seeded on nano-hydroxyapatite/collagen/poly(L-lactide).

    PubMed

    E, Ling-Ling; Xu, Wen-Huan; Feng, Lin; Liu, Yi; Cai, Dong-Qing; Wen, Ning; Zheng, Wen-Jie

    2016-06-01

    This study investigated the effects of estrogen on the bone regeneration potential of periodontal ligament stem cells (PDLSCs) derived from osteoporotic rats and seeded on a collagen-based composite scaffold [nano-hydroxyapatite/collagen/poly(L-lactide) (nHAC/PLA)]. For this purpose, 48 healthy 3‑month-old Sprague-Dawley female rats were divided into 2 groups as follows: the bilaterally ovariectomized (OVX) rats and sham‑operated rats. The PDLSCs were isolated at 3 months after surgery (by which time postmenopausal osteoporosis had developed). The effects of estrogen on the characteristics of these cells seeded in a culture plate and of the cells seeded on nHAC/PLA were then investigated. The PDLSC + nHAC/PLA constructs were implanted subcutaneously into the backs of severe combined immunodeficient (SCID) mice for 12 weeks in order to examine the role of estrogen in the bone formation ability of PDLSCs derived from osteoporotic rats. The results from methyl thiazolyl tetrazolium (MTT) assay revealed that the proliferation of the cells derived from the rats in the OVX group was significantly higher than that of the cells derived from the rats in the sham-operated group at the stage of logarithmic growth. The staining intensity of alkaline phosphatase (ALP) and the mineralization of the cells derived from the rats in the OVX group was significantly weaker than that of the cells from the rats in the sham-operated group. When the PDLSCs were seeded on nHAC/PLA, ALP activity, osteocalcin (OCN) secretion, mineral formation and the mRNA expression levels of ALP, OCN, estrogen receptor (ER)α and ERβ in the cells derived from the rats in the OVX group were markedly decreased. Treatment with 17β-estradiol (E2) significantly weakened the proliferative ability of the cells derived from the OVX group rats, and enhanced their osteogenic differentiation ability and the mRNA expression levels of ALP, OCN, ERα and ERβ. When the constructs were implanted

  13. Rapid biomimetic mineralization of collagen fibrils and combining with human umbilical cord mesenchymal stem cells for bone defects healing.

    PubMed

    Ye, Bihua; Luo, Xueshi; Li, Zhiwen; Zhuang, Caiping; Li, Lihua; Lu, Lu; Ding, Shan; Tian, Jinhuan; Zhou, Changren

    2016-11-01

    Collagen biomineralization is regulated by complicated interactions between the collagen matrix and non-collagenous extracellular proteins. Here, the use of sodium tripolyphosphate to simulate the templating functional motif of the C-terminal fragment of non-collagenous proteins is reported, and a low molecular weight polyacrylic acid served as a sequestration agent to stabilize amorphous calcium phosphate into nanoprecursors. Self-assembled collagen fibrils served as a fixed template for achieving rapid biomimetic mineralization in vitro. Results demonstrated that, during the mineralization process, intrafibrillar and extrafibrillar hydroxyapatite mineral with collagen fibrils formed and did so via bottom-up nanoparticle assembly based on the non-classical crystallization approach in the presence of these dual biomimetic functional analogues. In vitro human umbilical cord mesenchymal stem cell (hUCMSC) culture found that the mineralized scaffolds have a better cytocompatibility in terms of cell viability, adhesion, proliferation, and differentiation into osteoblasts. A rabbit femoral condyle defect model was established to confirm the ability of the n-HA/collagen scaffolds to facilitate bone regeneration and repair. The images of gross anatomy, MRI, CT and histomorphology taken 6 and 12weeks after surgery showed that the biomimetic mineralized collagen scaffolds with hUCMSCs can promote the healing speed of bone defects in vivo, and both of the scaffolds groups performing better than the bone defect control group. As new bone tissue formed, the scaffolds degraded and were gradually absorbed. All these results demonstrated that both of the scaffolds and cells have better histocompatibility. PMID:27523994

  14. Quantitative analysis on orientation of human bone integrated with midpalatal implant by micro X-ray diffractometer

    NASA Astrophysics Data System (ADS)

    Murata, Masaru; Akazawa, Toshiyuki; Yuasa, Toshihiro; Okayama, Miki; Tazaki, Junichi; Hanawa, Takao; Arisue, Makoto; Mizoguchi, Itaru

    2012-12-01

    A midpalatal implant system has been used as the unmoved anchorage for teeth movement. An 18-year-old male patient presented with reversed occlusion and was diagnosed as malocclusion. A pure titanium fixture (lengths: 4 mm, diameter: 3.3 mm, Orthosystem®, Institute Straumann, Switzerland) was implanted into the palatal bone of the patient as the orthodontic anchorage. The implant anchorage was connected with the upper left and right first molars, and had been used for 3 years. After dynamic treatments, the titanium fixture connected with bone was removed surgically, fixed in formalin solution, and embedded in resin. Specimens were cut along the frontal section of face and the direction of longitudinal axis of the implant, stained, and observed histologically. The titanium fixture was integrated directly with compact bone showing cortical bone-like structure such as lamella and osteon. In addition, to qualitatively characterize the implant-supported human bone, the crystallinity and orientation of hydroxyapatite (HAp) phase were evaluated by the microbeam X-ray diffraction analysis. Preferential alignment of c-axis of HAp crystals was represented by the relative intensity ratio of (0 0 2)-face diffraction peak to (3 1 0)-face one. The values decreased monotonously along the direction of the lateral stress from the site near the implant thread to the distant site in all horizontal lines of the map. These results indicated that the X-ray images for the intensity of c-face in HAp revealed functionally graded distribution of cortical bone quality. The micro-scale measurements of HAp structure could be a useful method for evaluating the mechanical stress distribution in human hard tissues.

  15. Adsorption of Amorphous Silica Nanoparticles onto Hydroxyapatite Surfaces Differentially Alters Surfaces Properties and Adhesion of Human Osteoblast Cells.

    PubMed

    Kalia, Priya; Brooks, Roger A; Kinrade, Stephen D; Morgan, David J; Brown, Andrew P; Rushton, Neil; Jugdaohsingh, Ravin

    2016-01-01

    Silicon (Si) is suggested to be an important/essential nutrient for bone and connective tissue health. Silicon-substituted hydroxyapatite (Si-HA) has silicate ions incorporated into its lattice structure and was developed to improve attachment to bone and increase new bone formation. Here we investigated the direct adsorption of silicate species onto an HA coated surface as a cost effective method of incorporating silicon on to HA surfaces for improved implant osseointegration, and determined changes in surface characteristics and osteoblast cell adhesion. Plasma-sprayed HA-coated stainless steel discs were incubated in silica dispersions of different concentrations (0-42 mM Si), at neutral pH for 12 h. Adsorbed Si was confirmed by XPS analysis and quantified by ICP-OES analysis following release from the HA surface. Changes in surface characteristics were determined by AFM and measurement of surface wettability. Osteoblast cell adhesion was determined by vinculin plaque staining. Maximum Si adsorption to the HA coated disc occurred after incubation in the 6 mM silica dispersion and decreased progressively with higher silica concentrations, while no adsorption was observed with dispersions below 6 mM Si. Comparison of the Si dispersions that produced the highest and lowest Si adsorption to the HA surface, by TEM-based analysis, revealed an abundance of small amorphous nanosilica species (NSP) of ~1.5 nm in diameter in the 6 mM Si dispersion, with much fewer and larger NSP in the 42 mM Si dispersions. 29Si-NMR confirmed that the NSPs in the 6 mM silica dispersion were polymeric and similar in composition to the larger NSPs in the 42 mM Si dispersion, suggesting that the latter were aggregates of the former. Amorphous NSP adsorbed from the 6 mM dispersion on to a HA-coated disc surface increased the surface's water contact angle by 53°, whereas that adsorbed from the 42 mM dispersion decreased the contact angle by 18°, indicating increased and decreased

  16. Adsorption of Amorphous Silica Nanoparticles onto Hydroxyapatite Surfaces Differentially Alters Surfaces Properties and Adhesion of Human Osteoblast Cells

    PubMed Central

    Kalia, Priya; Brooks, Roger A.; Kinrade, Stephen D.; Morgan, David J.; Brown, Andrew P.; Rushton, Neil; Jugdaohsingh, Ravin

    2016-01-01

    Silicon (Si) is suggested to be an important/essential nutrient for bone and connective tissue health. Silicon-substituted hydroxyapatite (Si-HA) has silicate ions incorporated into its lattice structure and was developed to improve attachment to bone and increase new bone formation. Here we investigated the direct adsorption of silicate species onto an HA coated surface as a cost effective method of incorporating silicon on to HA surfaces for improved implant osseointegration, and determined changes in surface characteristics and osteoblast cell adhesion. Plasma-sprayed HA-coated stainless steel discs were incubated in silica dispersions of different concentrations (0–42 mM Si), at neutral pH for 12 h. Adsorbed Si was confirmed by XPS analysis and quantified by ICP-OES analysis following release from the HA surface. Changes in surface characteristics were determined by AFM and measurement of surface wettability. Osteoblast cell adhesion was determined by vinculin plaque staining. Maximum Si adsorption to the HA coated disc occurred after incubation in the 6 mM silica dispersion and decreased progressively with higher silica concentrations, while no adsorption was observed with dispersions below 6 mM Si. Comparison of the Si dispersions that produced the highest and lowest Si adsorption to the HA surface, by TEM-based analysis, revealed an abundance of small amorphous nanosilica species (NSP) of ~1.5 nm in diameter in the 6 mM Si dispersion, with much fewer and larger NSP in the 42 mM Si dispersions. 29Si-NMR confirmed that the NSPs in the 6 mM silica dispersion were polymeric and similar in composition to the larger NSPs in the 42 mM Si dispersion, suggesting that the latter were aggregates of the former. Amorphous NSP adsorbed from the 6 mM dispersion on to a HA-coated disc surface increased the surface’s water contact angle by 53°, whereas that adsorbed from the 42 mM dispersion decreased the contact angle by 18°, indicating increased and decreased

  17. Human temporal bone findings in acquired hypothyroidism.