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1

Primary lymphomas of bone.  

PubMed

Primary bone lymphoma is rare. The majority of cases are diffuse large B-cell non-Hodgkin's lymphomas. Classification, staging, and treatment are controversial. The relatively small number of cases has led to many case reports and series describing institutional experiences but precludes the use of randomized clinical trials to address the question of optimal management. This article will review clinical and radiologic presentations, diagnostic techniques, and histologic characteristics. Most important, it will present what limited information we do have regarding effective treatment options for this unusual type of lymphoma. PMID:16231853

Gill, Paula; Wenger, Doris E; Inwards, David J

2005-09-01

2

Primary bone tumors of adulthood  

PubMed Central

Imaging plays a crucial role in the evaluation of primary bone tumors in adults. Initial radiographic evaluation is indicated in all cases with suspected primary bone tumors. Radiographs are useful for providing the diagnosis, a short list of differential diagnosis or at least indicating the degree of aggressiveness of the lesion. More detailed information about the lesion, such as cortical destruction or local spread, can be obtained using cross-sectional imaging techniques such as computed tomography and magnetic resonance imaging. This article discusses the characteristic features of the more common primary bone tumors of adulthood, and also the pre-treatment evaluation and staging of these lesions using imaging techniques.

Teo, Harvey E L; Peh, Wilfred C G

2004-01-01

3

Bone disease in primary hyperparathyrodism  

PubMed Central

Nowadays, primary hyperparathyroidism (PHPT) is mostly a mild disease. Overt skeletal manifestations are rare but decreased bone mineral density (BMD) can still be demonstrated. Even in mild cases, excess parathyroid hormone (PTH) increases bone turnover leading to bone loss particularly at cortical sites. Conversely, a relative preservation of cancellous bone has been shown by histomorphometric analyses and advanced imaging techniques. An increased fracture rate has been demonstrated in untreated patients with PHPT at peripheral sites and in the spine. Parathyroidectomy (PTx) is the definitive cure for PHPT. With the restoration of normal PTH, bone resorption is quickly tapered down, while bone formation proceeds at the level of bone multicellular units, which were activated prior to PTx. The rapid refilling of the enlarged remodeling space and the subsequent matrix mineralization will result in an increase in BMD at sites rich in trabecular bone, such as lumbar spine and hip, which mainly occurs during the first 6–12 months after PTx. Cortical bone is less responsive to PTX because of the low rate of bone turnover, but sensible increases in BMD at the distal third of the radius can be observed in the long term. PTx seems to decrease the risk of fractures but more data are needed before a definitive conclusion on this important matter can be reached. Treatment with bisphosphonates can be considered for patients with low BMD who do not undergo PTx. Two-year treatment with alendronate has been shown to decrease bone turnover markers and increase BMD at the lumbar spine and hip, but not at the distal radius. Cinacalcet stably decreased serum calcium levels across a broad range of PHPT severity, but no change in BMD occurred in patients treated for up to 5.5 years.

Cianferotti, Luisella; Cetani, Filomena

2012-01-01

4

Primary bone marrow oedema syndromes.  

PubMed

MRI scanning in patients with rheumatological conditions often shows bone marrow oedema, which can be secondary to inflammatory, degenerative, infective or malignant conditions but can also be primary. The latter condition is of uncertain aetiology and it is also uncertain whether it represents a stage in the progression to osteonecrosis in some patients. Patients with primary bone marrow oedema usually have lower limb pain, commonly the hip, knee, ankle or feet. The diagnosis is one of exclusion with the presence of typical MRI findings. Treatment is usually conservative and includes analgesics and staying off the affected limb. The natural history is that of gradual resolution of symptoms over a number of months. Evidence for medical treatment is limited, but open-label studies suggest bisphosphonates may help in the resolution of pain and improve radiological findings. Surgical decompression is usually used as a last resort. PMID:24080251

Patel, Sanjeev

2014-05-01

5

Primary extra nodal Hodgkin disease: Bone presentation  

PubMed Central

Summary Background Extra nodal and extra lymphatic propagation of Hodgkin’s disease is a characteristic of the fourth stage of disease when the organs are affected. Primary appearances of the disease outside the lymph node is a rare event. Therefore, it makes diagnostic problem. Skeletal system is possible localization of primary extra nodal Hodgkin’s disease. Case Report Women, 42-years-old, was admitted to hospital because of swelling and pain in the right shoulder. After imaging and histological examination diagnosed Hodgkin’s nodular sclerosing histological subtype disease has been established. The patient starts to receive chemotherapy. Conclusions Primary extra nodal Hodgkin’s disease of bone is manifested with painful swelling in geared area. Imaging method shows destruction of the affected bone, with swelling of the soft tissues. Propagation in soft tissue is not accompanied by their destruction, but rather manifested swelling of the surrounding soft tissue.

Nikolica, Goran; Badnjar, Zorka; Cadjenovic, Tanja; Raceta-Masic, Dijana

2014-01-01

6

Is miniscrew primary stability influenced by bone density?  

PubMed

Primary stability is absence of mobility in the bone bed after mini-implant placement and depends on bone quality among other factors. Bone quality is a subjective term frequently considered as bone density. The aim of this preliminary study was to evaluate bone density in two bovine pelvic regions and verify the primary stability of miniscrews inserted into them. Forty bone blocks were extracted from bovine pelvic bones, 20 from iliac and 20 from pubic bone, all of them containing cortical bone about 1 mm thick. Half of the sections extracted from each bone were designated for histological evaluation of bone density (trabecular bone area - TBA) and the other half for bone mineral density (BMD) evaluation by means of central dual-energy X-ray absorptiometry (DEXA). Then, twenty self-drilling miniscrews (INP®, São Paulo, Brazil) 1.4 mm in diameter and 6 mm long were inserted into the bone blocks used for BMD evaluation. Peak implant insertion torque (IT) and pull-out strength (PS) were used for primary stability evaluation. It was found that iliac and pubic bones present different bone densities, iliac bone being less dense considering BMD and TBA values (P > 0.05). However, the miniscrew primary stability was not different when varying the bone type (P < 0.05). IT and PS were not influenced by these differences in bone density when cortical thickness was about 1 mm thick. PMID:22031056

Marquezan, Mariana; Souza, Margareth Maria Gomes de; Araújo, Mônica Tirre de Souza; Nojima, Lincoln Issamu; Nojima, Matilde da Cunha Gonçalves

2011-01-01

7

Localization of primary cilia in mouse retina.  

PubMed

The primary cilia are considered as "cellular antennae" that sense and interchange information with the extracellular environment. Nearly all mammalian cells have a single primary cilium. In the retina, the outer segment of the photoreceptor is known to be a specialized form of primary cilium, but studies on cilia in other layers of the retina are scarce. In this study, we investigated the expression of primary cilia in the whole thickness of the mouse retina using immunofluorescence with three different ciliary markers: Arl13b, acetylated ?-tubulin and adenylyl cyclase III. Our results show positive reactions in the photoreceptor layer, outer plexiform layer and ganglion cell layer, which might suggest the possible presence of primary cilia in these areas, but we could not directly prove the strand-like shape of cilia in those areas. In the outer plexiform layer, all three markers showed intense staining along the neuronal synapses, which suggests that the neuronal processes themselves might share the features of cilia. PMID:23608602

Kim, Yong-Kyu; Kim, Jin Hyoung; Yu, Young Suk; Ko, Hyuk Wan; Kim, Jeong Hun

2013-10-01

8

The Primary Cilium as a Novel Extracellular Sensor in Bone  

PubMed Central

Mechanically induced adaptation of bone is required to maintain a healthy skeleton and defects in this process can lead to dramatic changes in bone mass, resulting in bone diseases such as osteoporosis. Therefore, understanding how this process occurs could yield novel therapeutics to treat diseases of excessive bone loss or formation. Over the past decade the primary cilium has emerged as a novel extracellular sensor in bone, being required to transduce changes in the extracellular mechanical environment into biochemical responses regulating bone adaptation. In this review, we introduce the primary cilium as a novel extracellular sensor in bone; discuss the in vitro and in vivo findings of primary cilia based sensing in bone; explore the role of the primary cilium in regulating stem cell osteogenic fate commitment and finish with future directions of research and possible development of cilia targeting therapeutics to treat bone diseases.

Hoey, David A.; Chen, Julia C.; Jacobs, Christopher R.

2012-01-01

9

Mouse Models in Bone Marrow Transplantation and Adoptive Cellular Therapy  

PubMed Central

Mouse models of transplantation have been indispensable to the development of bone marrow transplantation (BMT). Their role in the generation of basic science knowledge is invaluable and is subject to discussion below. However, this article focuses on the direct role and relevance of mouse models towards the clinical development and advances in BMT and adoptive T-cell therapy for human diseases. The authors aim to present a thoughtful perspective on the pros and cons of mouse models while noting that despite imperfections these models are obligatory for the development of science-based medicine.

Arber, Caroline; Brenner, Malcolm K.; Reddy, Pavan

2014-01-01

10

Cellular lead toxicity and metabolism in primary and clonal osteoblastic bone cells  

SciTech Connect

A knowledge of bone lead metabolism is critical for understanding the toxicological importance of bone lead, as a toxicant both to bone cells and to soft tissues of the body, as lead is mobilized from large reservoirs in hard tissues. To further understand the processes that mediate metabolism of lead in bone, it is necessary to determine lead metabolism at the cellular level. Experiments were conducted to determine the intracellular steady-state {sup 210}Pb kinetics in cultures of primary and clonal osteoblastic bone cells. Osteoblastic bone cells obtained by sequential collagenase digestion of mouse calvaria or rat osteosarcoma (ROS 17/2.8) cells were labeled with {sup 210}Pb as 5 microM lead acetate for 20 hr, and kinetic parameters were determined by measuring the efflux of {sup 210}Pb from the cells over a {sup 210}-min period. The intracellular metabolism of {sup 210}Pb was characterized by three kinetic pools of {sup 210}Pb in both cell types. Although the values of these parameters differed between the primary osteoblastic cells and ROS cells, the profile of {sup 210}Pb was remarkably similar in both cell types. Both types exhibited one large, slowly exchanging pool (S3), indicative of mitochondrial lead. These data show that primary osteoblastic bone cells and ROS cells exhibit similar steady-state lead kinetics, and intracellular lead distribution. These data also establish a working model of lead kinetics in osteoblastic bone cells and now permit an integrated view of lead kinetics in bone.

Long, G.J.; Rosen, J.F.; Pounds, J.G. (Brookhaven National Laboratory, Upton, NY (USA))

1990-02-01

11

Bone histomorphometry and serum bone gla-protein in the diagnosis of primary hyperparathyroidism  

Microsoft Academic Search

The diagnosis of primary hyperparathyroidism (PHP) may be difficult, especially in the case of asymptomatic hypercalcemia. Since bone is the major target organ of parathyroid hormone (PTH), the hypersecretion of PTH in patients with PHP can be assessed by bone biopsy and by measured markers of bone turnover. In a first study, a transiliac bone biopsy was performed in 184

Pierre D. Delmas; Pierre J. Meunier; E. Faysse; E. C. Saubier

1986-01-01

12

Cranial bone morphometric study among mouse strains  

Microsoft Academic Search

BACKGROUND: Little is known about the molecular mechanism which regulates how the whole cranium is shaped. Mouse models currently available for genetic research include several hundreds of unique inbred strains and genetically engineered mutants. By cross comparing their genomic structures, we can elucidate the cause of any differences in the phenotype between two strains. The craniometry of subspecies, or closely

Minoru Kawakami; Ken-ichi Yamamura

2008-01-01

13

How tough is brittle bone? Investigating osteogenesis imperfecta in mouse bone.  

PubMed

The multiscale hierarchical structure of bone is naturally optimized to resist fractures. In osteogenesis imperfecta, or brittle bone disease, genetic mutations affect the quality and/or quantity of collagen, dramatically increasing bone fracture risk. Here we reveal how the collagen defect results in bone fragility in a mouse model of osteogenesis imperfecta (oim), which has homotrimeric ?1(I) collagen. At the molecular level, we attribute the loss in toughness to a decrease in the stabilizing enzymatic cross-links and an increase in nonenzymatic cross-links, which may break prematurely, inhibiting plasticity. At the tissue level, high vascular canal density reduces the stable crack growth, and extensive woven bone limits the crack-deflection toughening during crack growth. This demonstrates how modifications at the bone molecular level have ramifications at larger length scales affecting the overall mechanical integrity of the bone; thus, treatment strategies have to address multiscale properties in order to regain bone toughness. In this regard, findings from the heterozygous oim bone, where defective as well as normal collagen are present, suggest that increasing the quantity of healthy collagen in these bones helps to recover toughness at the multiple length scales. © 2014 American Society for Bone and Mineral Research. PMID:24420672

Carriero, Alessandra; Zimmermann, Elizabeth A; Paluszny, Adriana; Tang, Simon Y; Bale, Hrishikesh; Busse, Bjorn; Alliston, Tamara; Kazakia, Galateia; Ritchie, Robert O; Shefelbine, Sandra J

2014-06-01

14

The construction of an in vitro three-dimensional hematopoietic microenvironment for mouse bone marrow cells employing porous carriers  

Microsoft Academic Search

Spatial development of mouse bone marrow cellsemploying porous carriers was investigated in order todesign a bioreactor with a three-dimensionalhematopoietic microenvironment. Three types of porouscarriers were used for examining the spatialdevelopment of anchorage-dependent primary stromalcells as feeder cells. Stromal cells were found tospread well at a high density on a polyester nonwovendisc carrier (Fibra cel (FC)) under a scanningelectron microscope, while

Yoshiaki Tomimori; Mutsumi Takagi; Toshiomi Yoshida

2000-01-01

15

Identification of Clonogenic Common Lymphoid Progenitors in Mouse Bone Marrow  

Microsoft Academic Search

The existence of a common lymphoid progenitor that can only give rise to T cells, B cells, and natural killer (NK) cells remains controversial and constitutes an important gap in the hematopoietic lineage maps. Here, we report that the Lin?IL-7R+Thy-1?Sca-1loc-Kitlo population from adult mouse bone marrow possessed a rapid lymphoid-restricted (T, B, and NK) reconstitution capacity in vivo but completely

Motonari Kondo; Irving L. Weissman; Koichi Akashi

1997-01-01

16

Porphyromonas gingivalis infection increases osteoclastic bone resorption and osteoblastic bone formation in a periodontitis mouse model  

PubMed Central

Background Porphyromonas gingivalis has been shown to invade osteoblasts and inhibit their differentiation and mineralization in vitro. However, it is unclear if P. gingivalis can invade osteoblasts in vivo and how this would affect alveolar osteoblast/osteoclast dynamics. This study aims to answer these questions using a periodontitis mouse model under repetitive P. gingivalis inoculations. Methods For 3-month-old BALB/cByJ female mice, 109 CFU of P. gingivalis were inoculated onto the gingival margin of maxillary molars 4 times at 2-day intervals. After 2 weeks, another 4 inoculations at 2-day intervals were applied. Calcein was injected 7 and 2 days before sacrificing animals to label the newly formed bone. Four weeks after final inoculation, mice were sacrificed and maxilla collected. Immunohistochemistry, micro-CT, and bone histomorphometry were performed on the specimens. Sham infection with only vehicle was the control. Results P. gingivalis was found to invade gingival epithelia, periodontal ligament fibroblasts, and alveolar osteoblasts. Micro-CT showed alveolar bone resorption and significant reduction of bone mineral density and content in the infected mice compared to the controls. Bone histomorphometry showed a decrease in osteoblasts, an increase in osteoclasts and bone resorption, and a surprisingly increased osteoblastic bone formation in the infected mice compared to the controls. Conclusions P. gingivalis invades alveolar osteoblasts in the periodontitis mouse model and cause alveolar bone loss. Although P. gingivalis appears to suppress osteoblast pool and enhance osteoclastic bone resorption, the bone formation capacity is temporarily elevated in the infected mice, possibly via some anti-microbial compensational mechanisms.

2014-01-01

17

Primary aneurysmal bone cyst of coronoid process  

Microsoft Academic Search

BACKGROUND: Aneurysmal bone cysts are relatively uncommon in the facial skeleton. These usually affect the mandible but origin from the coronoid process is even rarer. To the best of our knowledge, this is the first reported case of a coronoid process aneurysmal bone cyst presenting as temporal fossa swelling. CASE PRESENTATION: A 17 year old boy presented with a progressively

Amit Goyal; Isha Tyagi; Rajan Syal; Tanu Agrawal; Manoj Jain

2006-01-01

18

Smooth muscle actin expression in primary bone tumours.  

PubMed

Alpha isoform of smooth muscle actin (SMA) expression has been reported in giant cell tumour of bone (GCTB) and other benign and malignant bone tumours, but the pattern of SMA expression and the precise nature of SMA-expressing cells in these lesions is uncertain. We determined by immunohistochemistry the expression of SMA and other muscle and vascular markers in normal bone, GCTB and a wide range of primary benign and malignant bone tumours. Cultured stromal cells of GCTB, chondroblastoma (CB), and aneurysmal bone cyst (ABC) were also analysed for SMA expression. SMA was only noted in blood vessels in normal bone. SMA was expressed by mononuclear stromal cells (MSC) cultured from GCTB, ABC and CB. SMA was strongly and diffusely expressed by MSC in non-ossifying fibroma, fibrous dysplasia, and "brown tumour" of hyperparathyroidism. SMA expression was also noted in GCTB, ABC, CB, chondromyxoid fibroma, malignant fibrous histiocytoma of bone and osteosarcoma. Little or no SMA was noted in Langerhans cell histiocytosis, simple bone cyst, Ewing's sarcoma, osteoblastoma, osteoid osteoma, enchondroma, osteochondroma, chondrosarcoma, myeloma, lymphoma, chordoma and adamantinoma. Our findings show that there is differential SMA expression in primary bone tumours and that identifying the presence or absence of SMA is useful in the differential diagnosis of these lesions. The nature of SMA-expressing cells in bone tumours is uncertain but they are negative for desmin and caldesmon and could represent either myofibroblasts or perivascular cells, such as pericytes. PMID:22543453

Hemingway, F; Kashima, T G; Mahendra, G; Dhongre, A; Hogendoorn, P C W; Mertens, F; Athanasou, N A

2012-05-01

19

[Distribution of compact bone mesenchymal stem cells in lung tissue and bone marrow of mouse].  

PubMed

This study was aimed to investigate the distribution of compact bone mesenchymal stem cells(MSC) marked with lentiviral plasmid pGC FU-RFP-LV in lung tissue and bone marrow of mouse. The MSC were infected by lentivirus with infection efficiency 78%, the infected MSC were injected into BALB/c mice via tail veins in concentration of 1×10(6) /mouse. The mice were randomly divided into 4 group according to 4 time points as 1, 2, 5 and 7 days. The lung tissue and bone marrow were taken and made of frozen sections and smears respectively in order to observed the distributions of MSC. The results indicated that the lentiviral infected MSC displayed phenotypes and biological characteristics which conformed to MSC by immunophenotyping analysis and induction differentiation detection. After the MSC were infected with optimal viral titer MOI = 50, the cell growth no significantly changed; the fluorescent microscopy revealed that the distributions of MSC in bone marrow on day 1, 2, 5 and 7 were 0.50 ± 0.20, 0.67 ± 0.23, 0.53 ± 0.14, 0.33 ± 0.16; those in lung tissue were 0.55 ± 0.15, 0.47 ± 0.13, 0.29 ± 0.13, 0.26 ± 0.08. It is concluded that the distribution of MSC in lung tissue reaches a peak on day 1, while distribution of MSC in bone marrow reaches a peak on day 2. The distribution of mouse MSC relates with RFP gene expression and implantation of MSC in lung tissue and bone marrow. PMID:24598672

Wang, Rui-Ping; Wu, Ren-Na; Guo, Yu-Qing; Zhang, Bin; Chen, Hu

2014-02-01

20

FDG PET/CT of Primary Bone Tumors.  

PubMed

OBJECTIVE. Numerous primary bone tumors are encountered on (18)F-FDG PET/CT, and many are FDG avid. The degree of FDG uptake in bone tumors does not necessarily reflect malignant potential. In conjunction with radiographs, evaluation of morphologic characteristics on the CT portion of PET/CT scans is important for characterization of the lesions. FDG PET/CT has been found to be useful for staging and has also been found to reflect prognosis in some primary bone malignancies. The purpose of this article is to familiarize the reader with topics regarding FDG PET/CT and both malignant and benign primary bone tumors. CONCLUSION. FDG uptake alone is not adequate for characterizing primary bone tumors, and morphologic evaluation is an important factor in the interpretation of PET/CT scans. After diagnosis, FDG avidity and morphologic features can play an important role in staging and determining response to therapy. On completion of this article, readers should have an improved ability to evaluate the FDG uptake and CT morphologic features of malignant and benign primary bone tumors. Readers should also have a better understanding of the potential role of FDG PET/CT in the management of patients with osteosarcoma and Ewing sarcoma. PMID:24848845

Costelloe, Colleen M; Chuang, Hubert H; Madewell, John E

2014-06-01

21

Mouse bone marrow cytogenetic damage produced by residues of tequila.  

PubMed

Five concentrations (50-860 mg/kg) of residues obtained after distillation and lyophilization of commercial tequila were injected into mice for evaluation of chromosome aberrations, sister-chromatid exchanges, and proliferation kinetics in mouse bone marrow cells. Appropriate positive and negative controls were included. Our results showed significant dose-related increases of chromosomal aberrations starting at 50 mg/kg and for sister-chromatid exchanges at 430 mg/kg. Cellular proliferation kinetics showed no alterations. With these data we demonstrated that the residues of tequila are genotoxic in vivo. PMID:2345552

Madrigal-Bujaidar, E; Rojas, A; Ramos, A; Rosas, E; Díaz Barriga-Arceo, S

1990-06-01

22

BONE FORMATION INDUCED IN MOUSE THIGH BY CULTURED HUMAN CELLS  

PubMed Central

Cultured FL human amnion cells injected intramuscularly into cortisone-conditioned mice proliferate to form discrete nodules which become surrounded by fibroblasts. Within 12 days, fibroblastic zones differentiate into cartilage which calcifies to form bone. Experiments were conducted to test the hypothesis that FL cells behave as an inductor of bone formation. In the electron microscope, FL cells were readily distinguished from surrounding fibroblasts. Transitional forms between the two cell types were not recognized. Stains for acid mucopolysaccharides emphasized the sharp boundary between metachromatic fibroblastic and cartilaginous zones and nonmetachromatic FL cells. 35S was taken up preferentially by fibroblasts and chondrocytes and then deposited extracellularly in a manner suggesting active secretion of sulfated mucopolysaccharides. FL cells showed negligible 35S utilization and secretion. FL cells, labeled in vitro with thymidine-3H, were injected and followed radioautographically, during bone formation. Nuclear label of injected FL cells did not appear in adjacent fibroblasts in quantities sufficient to indicate origin of the latter from FL cells. The minimal fibroblast nuclear labeling seen may represent reutilization of label from necrotic FL cells. It is suggested that FL cells injected into the mouse thigh induced cartilage and bone formation by host fibroblasts.

Anderson, H. Clarke; Coulter, P. R.

1967-01-01

23

Primary fibrosarcoma of bone. A clinicopathologic study of 130 patients.  

PubMed

One hundred thirty patients with histologically verified primary fibrosarcoma of bone, unassociated with any pre-existent benign bone condition, were treated at Memorial Sloan-Kettering Cancer Center between 1918 and 1973. This series of cases represents approximately 5% of primary malignant bone tumors treated in our institution. Eighty-nine of the lesions were medullary or central in location, and 41 were periosteal or peripheral. There was a nearly equal sex distribution, and a mean age of 38 years ranging from 4 to 83 years. This lesion exhibited a strong predilection for long bones, with the most common location being the femur (43 cases), humerus (16 cases), and tibia (12 cases). In 19 instances, bones of the head and neck area were the primary sites. The roentgenographic differential diagnoses included osteolytic osteogenic sarcoma, malignant giant cell tumor, metastatic carcinoma, or solitary plasma cell myeloma. Major ablative surgery was the primary method of therapy. Amputation was performed, yielding the best curative results in high-grade tumors, while radical local excision sufficed for most low-grade periosteal fibrosarcomas. Thirty-four percent of the patients survived 5 years (27% medullary and 52% periosteal), while 28% were alive after 10 years (20% medullary and 48% periosteal). These survival rates provide further evidence that fibrosarcoma of bone is a distinct clinicopathologic entity and not a variant of osteosarcoma, which carries a much poorer 5-year survival rate of approximately 17%. PMID:1053940

Huvos, A G; Higinbotham, N L

1975-03-01

24

Cementless fixation for primary segmental bone tumor endoprostheses.  

PubMed

To combat the high incidence of aseptic loosening for young patients and for patients with failed implants after resection for bone tumors, intramedullary cementless fixation of massive tumor implants was investigated. These implants consist of a hydroxyapatite coated titanium stem. To date, 47 of these prostheses have been inserted for the treatment of primary bone tumors. Radiographs indicate that the stems are osseointegrated. Radiolucent lines have not been seen between the implant and the bone. Bone remodeling changes have been observed. In several cases in which the implant was not seated properly on the transaction site, bone grew to the shoulder of the implant. Bone remodeling was particularly evident in stems that were coated over their entire surface. In these cases, the implant induced local bone resorption so that the bone around the midstream region became thinner, with resorption of cortical bone on the periosteal surface and maintenance of bone on the endosteal surface adjacent to the stem. This effect was attributed to stress shielding, and a three-dimensional finite element model using loading data obtained from a telemetry study indicated that, where the stem was bonded to the bone over the entire surface, stresses in the outer cortex became reduced. In the finite element model, reducing the region of hydroxyapatite coating to approximately 1/3 of the stem length reduced the extent of the low-stress area in the outer cortex. Subsequently, prostheses have been coated with hydroxyapatite over only approximately 1/3 of their stem. This method of fixing the massive endoprosthesis to the bone is thought to be successful in the short-term and offers an alternative to cemented fixation. PMID:10738431

Blunn, G W; Briggs, T W; Cannon, S R; Walker, P S; Unwin, P S; Culligan, S; Cobb, J P

2000-03-01

25

A practical guide to culturing mouse and human bone marrow stromal cells.  

PubMed

Bone marrow stromal cells (BMSCs, frequently also called MSCs) represent a cell population within the bone marrow, a subset of which contains multipotent stem cells. Their primary role is to produce and maintain both bone tissue and bone marrow microenvironment necessary for hematopoiesis. The latter is achieved by secreting a wide variety of different cytokines and growth factors, many of which also have a regulatory role in immune processes. BMSCs have recently been introduced into the field of immunobiology after their successful clinical use in GVHD was reported in 2004. Since then, numerous studies confirmed and expanded the knowledge on the immunosuppressive potential of BMSCs in various in vitro and in vivo models. Although the immunomodulatory capacity of BMSCs is well established, there are still many unanswered questions regarding the cytokines, chemokines, receptors, and molecular pathways that play a role in this effect. To study these cells and answer many of the questions, researchers must be able to reliably and reproducibly isolate, culture, and use these cells. Below a practical guide on how to culture and characterize mouse and human BMSCs, which can then be applied in various in vitro and in vivo assays, is provided. PMID:24510517

Nemeth, K; Mayer, B; Sworder, B J; Kuznetsov, S A; Mezey, E

2013-01-01

26

Trabecular bone structure in patients with primary hyperparathyroidism  

Microsoft Academic Search

To evaluate the effects of increased parathyroid hormone (PTH)-secretion on trabecular bone structure in patients with primary hyperparathyroidism (PHPT) we have analysed iliac crest biopsies from 84 patients (38 male, 46 female, age 20–85 years) with surgically proven PHPT by using quantitative histomorphometry. As there might be an influence of age or sex, subjects were divided into 4 subgroups according

Martin Vogel; Michael Hahn; Günter Delling

1995-01-01

27

[Limb-saving resection procedures in primary malignant bone tumors].  

PubMed

Thoroughly indicated and performed limb salvaging resections of primary malignant bone tumors can be as well affective as amputations and disarticulations. Possible limb salvaging operative procedures are described with regard to histodiagnosis grading, localisation, expansion of neoplasma as well as to the personality of the patient. PMID:6367249

Cotta, H; Rohe, K

1984-01-01

28

Bone formation following transplantation of genetically modified primary bone marrow stromal cells  

Microsoft Academic Search

Bone marrow stromal cells contain mesenchymal stem cells that can differentiate into a variety of mesenchymal tissues; in the presence of BMP-2, for example, they differentiate into osteoblasts. We constructed replication-deficient adenoviral vectors encoding human BMP-2 (BMP-2\\/Ad) or BMP-4 (BMP-4\\/Ad) and used them to transduce primary bone marrow stromal cells from the femurs of four-week-old female C3H mice, which then

Osamu Sugiyama; Hideo Orimo; Satoru Suzuki; Kazuo Yamashita; Hiromoto Ito; Takashi Shimada

2003-01-01

29

Cytogenetic risk assessment of etoposide from mouse bone marrow.  

PubMed

Increased clinical applications of the anticancer drug etoposide (a non-intercalative epipodophyllotoxin derivative) and the frequent induction of a second malignancy, particularly leukaemia, in post-etoposide-treated cancer survivors warrant detailed genotoxicity testing of etoposide. The genotoxicity test results available on etoposide are either primarily in in vitro test systems or in lower organisms after treatment with unusually high doses, or after chronic exposures, having little extrapolative value to humans. Therefore, a cytogenetic risk assessment study on etoposide in mouse in vivo was undertaken after a low dose (in accordance with the human therapeutic dose) single exposure. The cytogenetic toxicity of etoposide was assessed from bone marrow of mouse at three separate endpoints: chromosomal aberration and mitotic index studies at 24 h post-treatment and the micronucleus test (MNT) at 30 h post-treatment. The flame drying technique using colchicine, hypotonic sodium citrate, methanol-glacial acetic acid and Giemsa was followed for the preparation of slides for the metaphase chromosomal aberration and mitotic index studies and a simple technique was followed for the MNT. Although induction of chromosomal aberrations, excluding gaps, per 100 metaphases by 10 and 15 mg kg(-1) etoposide was not significant statistically, 20 mg kg(-1) of etoposide induced a significantly higher number of chromosomal aberrations in female (P < or = 0.01) and male (P < or = 0.05) mice. There was no significant change in the induced percentages of dividing cells by any of the doses of etoposide tested. The micronucleus induction also was not significant statistically with the lowest dose but it was significant in female (P mouse bone marrow after a single treatment with such low doses. However, the drug did not interfere with cell cycle progression. Although it is a DNA-non-intercalating agent, etoposide is known for its interference in the activity of DNA topoisomerase IIalpha enzyme, particularly in the proliferative cells where the concentration and activity of the enzyme are greater. This might be the reason for the induction of leukaemia in post-etoposide-treated cancer survivors. Therefore, it has become absolutely necessary to make etoposide target-specific, i.e. specific to the topoisomerase II enzymes of cancerous cells. PMID:15052606

Choudhury, Ramesh C; Palo, Anil K; Sahu, Prajyoti

2004-01-01

30

Incidental pedal manifestation of primary bone lymphoma: a case report.  

PubMed

Primary lymphoma of bone (PLB) is an uncommon entity and is extremely rare in the foot and ankle. In this case, PLB was identified from the bone specimen after a bunionectomy of the first and fifth metatarsals. The diagnosis was confirmed with pathologic analysis, genetic karyotyping, positron emission and computed tomography scans, and fluorescence in situ hybridization (FISH). We felt that reporting this case was essential due to the rarity of its pedal occurrence and the lack of preoperative signs or symptoms. PMID:24901590

DeLauro, Nicole M; Sharma, Shital; Shah, Nrupa; Ahmed, Imtiaz

2014-05-01

31

Primary cholesteatoma of petrous bone presenting as cervical fistula.  

PubMed

A unique case of primary cholesteatoma of the petrous bone is described. The patient presented with a cutaneous fistula of the anterior cervical triangle and was initially diagnosed and treated as sebaceous cyst or imflammatory lympha node for 3 months. Later examination showed white mass in the right external auditory canal and a severe conductive hearing loss. Radiological findings revealed a massive soft tissue consistent with cholesteatoma in the right petrous bone. The lesion was successfully managed with an endoscopy-assisted technique leaving the neurovascular structures in situ without operative complications. A review of the literature shows that only a few cases of cervical abscess associated with mastoid cholesteatoma have been reported. We also discussed the diagnosis, clinical features and surgical strategy for petrous bone cholesteatoma. PMID:19167849

Lin, Ying; Chen, Yang; Lu, Lian-Jun; Qiao, Li; Qiu, Jian-Hua

2009-08-01

32

Binocular integration and disparity selectivity in mouse primary visual cortex  

PubMed Central

Signals from the two eyes are first integrated in primary visual cortex (V1). In many mammals, this binocular integration is an important first step in the development of stereopsis, the perception of depth from disparity. Neurons in the binocular zone of mouse V1 receive inputs from both eyes, but it is unclear how that binocular information is integrated and whether this integration has a function similar to that found in other mammals. Using extracellular recordings, we demonstrate that mouse V1 neurons are tuned for binocular disparities, or spatial differences, between the inputs from each eye, thus extracting signals potentially useful for estimating depth. The disparities encoded by mouse V1 are significantly larger than those encoded by cat and primate. Interestingly, these larger disparities correspond to distances that are likely to be ecologically relevant in natural viewing, given the stereo-geometry of the mouse visual system. Across mammalian species, it appears that binocular integration is a common cortical computation used to extract information relevant for estimating depth. As such, it is a prime example of how the integration of multiple sensory signals is used to generate accurate estimates of properties in our environment.

Scholl, Benjamin; Burge, Johannes

2013-01-01

33

Inhibition of Drynariae Rhizoma extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts  

Microsoft Academic Search

In the traditional Korean medicine, Drynariae Rhizoma (DR) [Drynaria fortunei (kunze) J. Sm] has been reported as a good enhancer for bone healing. In this experiment, we investigate the effects of DR on bone resorption using the bone cells culture. Different concentrations of crude extract of DR were added to mouse bone cells culture. The mitochondria activity of the bone

Ji-Cheon Jeong; Sung-Koo Kang; Cheol-Ho Youn; Chang-Whan Jeong; Hyung-Min Kim; Young-Choon Lee; Young-Chae Chang; Cheorl-Ho Kim

2003-01-01

34

[A new method for isolating and culturing mouse bone marrow mesenchymal stem cells].  

PubMed

This study was purposed to establish a convenient and efficient method for isolating and culturing mouse bone marrow mesenchymal stem cells (MSC). The femurs and tibias of mouse were taken under sterile condition. MSC were isolated and cultured with flushing- out bone marrow or collagenase-digested bone fragment or bone marrow plus bone fragment. MSC colony number and size were compared. Immunophenotype and differentiation ability were tested to identify MSC. The results showed that colonies from bone marrow plus bone fragment group came out earliest and the colony number was 20 ± 4 at day 4; there were 11.5 ± 2.5 colonies in collagenase-digested bone fragment group and 9.5 ± 1.5 in flushing- out bone marrow group. The total cell yields of MSC after passaging showed best in bone marrow plus bone fragment group. Flow cytometry data showed the cultured cells expressed Sca-1, CD44 and CD29, not expressed pan-leukocyte surface marker CD45 and endothelial cell marker CD31. The isolated and cultured MSC could differentiate into osteoblast at the osteogenic differentiation condition, or adipocyte at adipogenic differentiation condition. It is concluded that the method of bone marrow plus bone fragment is convenient and efficient for isolating and culturing MSC. PMID:24370049

Yang, Yan-Mei; Li, Hong; Zhang, Lei; Dang, Rui-Jie; Li, Ping; Wang, Xiao-Yan; Zhu, Heng; Guo, Xi-Min; Zhang, Yi; Liu, Yuan-Lin; Mao, Ning; Jiang, Xiao-Xia; Wen, Ning

2013-12-01

35

Longitudinal assessment of in vivo bone dynamics in a mouse tail model of postmenopausal osteoporosis.  

PubMed

Recently, it has been shown that transient bone biology can be observed in vivo using time-lapse micro-computed tomography (?CT) in the mouse tail bone. Nevertheless, in order for the mouse tail bone to be a model for human disease, the hallmarks of any disease must be mimicked. The aim of this study was to investigate whether postmenopausal osteoporosis could be modeled in caudal vertebrae of C57Bl/6 mice, considering static and dynamic bone morphometry as well as mechanical properties, and to describe temporal changes in bone remodeling rates. Twenty C57Bl/6 mice were ovariectomized (OVX, n = 11) or sham-operated (SHM, n = 9) and monitored with in vivo ?CT on the day of surgery and every 2 weeks after, up to 12 weeks. There was a significant decrease in bone volume fraction for OVX (-35%) compared to SHM (+16%) in trabecular bone (P < 0.001). For OVX, high-turnover bone loss was observed, with the bone resorption rate exceeding the bone formation rate (P < 0.001). Furthermore there was a significant decrease in whole-bone stiffness for OVX (-16%) compared to SHM (+11%, P < 0.001). From these results we conclude that the mouse tail vertebra mimics postmenopausal bone loss with respect to these parameters and therefore might be a suitable model for postmenopausal osteoporosis. When evaluating temporal changes in remodeling rates, we found that OVX caused an immediate increase in bone resorption rate (P < 0.001) and a delayed increase in bone formation rate (P < 0.001). Monitoring transient bone biology is a promising method for future research. PMID:22159822

Lambers, Floor M; Kuhn, Gisela; Schulte, Friederike A; Koch, Kathleen; Müller, Ralph

2012-02-01

36

In vitro organoid culture of primary mouse colon tumors  

PubMed Central

Several human and murine colon cancer cell lines have been established, physiologic integrity of colon tumors such as multiple cell layers, basal-apical polarity, ability to differentiate, and anoikis are not maintained in colon cancer derived cell lines. The present study demonstrates a method for culturing primary mouse colon tumor organoids adapted from Sato T et al. 1, which retains important physiologic features of colon tumors. This method consists of mouse colon tumor tissue collection, adjacent normal colon epithelium dissociation, colon tumor cells digestion into single cells, embedding colon tumor cells into matrigel, and selective culture based on the principle that tumor cells maintain growth on limiting nutrient conditions compared to normal epithelial cells. The primary tumor organoids if isolated from genetically modified mice provide a very useful system to assess tumor autonomous function of specific genes. Moreover, the tumor organoids are amenable to genetic manipulation by virus meditated gene delivery; therefore signaling pathways involved in the colon tumorigenesis could also be extensively investigated by overexpression or knockdown. Primary tumor organoids culture provides a physiologic relevant and feasible means to study the mechanisms and therapeutic modalities for colon tumorigenesis.

Shah, Yatrik M.

2014-01-01

37

Compartmental Bone Morphometry in the Mouse Femur: Reproducibility and Resolution Dependence of Microtomographic Measurements  

Microsoft Academic Search

Microcomputed tomography (?CT) is widely used for nondestructive bone phenotyping in small animals, especially in the mouse. Here, we investigated the reproducibility and resolution dependence of ?CT analysis of microstructural parameters in three different compartments in the mouse femur. Reproducibility was assessed with respect to precision error (PE%CV) and intraclass correlation coefficient (ICC). We examined 14 left femurs isolated postmortem

T. Kohler; M. Beyeler; D. Webster; R. Müller

2005-01-01

38

MRI detection of early bone metastases in B16 mouse melanoma models  

PubMed Central

Bone metastasis causes significant morbidity in cancer patients, including bone pain, pathologic fractures, nerve compression syndrome, and hypercalcemia. Animal models are utilized to study the pathogenesis of skeletal metastases and to evaluate potential therapeutic agents. Previously published methods for imaging bone metastasis in rodent models have focused on identifying advanced stage metastasis using simple X-rays. Here we report MRI as a method for detecting early bone metastases in mouse models in vivo. B16 mouse melanoma cells were injected into the left cardiac ventricle of C57BL/6 mice and magnetic resonance (MR) images were obtained of the left leg following the development of metastatic disease, when tumor associated bone destruction was histologically present but not visible by X-ray. T1 and T2 relaxation times of bone marrow were measured in healthy control mice and B16 melanoma tumor-bearing mice. Mean T2 values for normal marrow were 28 ms (SD 5) and for diseased bone marrow were 41 ms (SD 3). T2 relaxation time of diseased bone marrow is significantly longer than that of normal bone marrow (P < 0.0001) and can be used as a marker of early bone metastases. These studies demonstrate that MR imaging can detect bone marrow metastases in small animals prior to development of cortical bone loss identified by X-ray.

Gauvain, Karen M.; Garbow, Joel R.; Song, Sheng-Kwei; Hirbe, Angela C.; Weilbaecher, Katherine

2009-01-01

39

The relevance of mouse models for investigating age-related bone loss in humans.  

PubMed

Mice are increasingly used for investigation of the pathophysiology of osteoporosis because their genome is easily manipulated, and their skeleton is similar to that of humans. Unlike the human skeleton, however, the murine skeleton continues to grow slowly after puberty and lacks osteonal remodeling of cortical bone. Yet, like humans, mice exhibit loss of cancellous bone, thinning of cortical bone, and increased cortical porosity with advancing age. Histologic evidence in mice and humans alike indicates that inadequate osteoblast-mediated refilling of resorption cavities created during bone remodeling is responsible. Mouse models of progeria also show bone loss and skeletal defects associated with senescence of early osteoblast progenitors. Additionally, mouse models of atherosclerosis, which often occurs in osteoporotic participants, also suffer bone loss, suggesting that common diseases of aging share pathophysiological pathways. Knowledge of the causes of skeletal fragility in mice should therefore be applicable to humans if inherent limitations are recognized. PMID:23689830

Jilka, Robert L

2013-10-01

40

Curcumin analogue UBS109 prevents bone loss in breast cancer bone metastasis mouse model: involvement in osteoblastogenesis and osteoclastogenesis.  

PubMed

Bone metastasis of breast cancer typically leads to osteolysis, which causes severe pathological bone fractures and hypercalcemia. Bone homeostasis is skillfully regulated through osteoblasts and osteoclasts. Bone loss with bone metastasis of breast cancer may be due to both activation of osteoclastic bone resorption and suppression of osteoblastic bone formation. This study was undertaken to determine whether the novel curcumin analogue UBS109 has preventive effects on bone loss induced by breast cancer cell bone metastasis. Nude mice were inoculated with breast cancer MDA-MB-231 bone metastatic cells (10(6) cells/mouse) into the head of the right and left tibia. One week after inoculation, the mice were treated with control (vehicle), oral administration (p.o.) of UBS109 (50 or 150 mg/kg body weight), or intraperitoneal administration (i.p.) of UBS109 (10 or 20 mg/kg body weight) once daily for 5 days per week for 7 weeks. After UBS109 administration for 7 weeks, hind limbs were assessed using an X-ray diagnosis system and hematoxylin and eosion staining to determine osteolytic destruction. Bone marrow cells obtained from the femurs and tibias were cultured to estimate osteoblastic mineralization and osteoclastogenesis ex vivo and in vitro. Remarkable bone loss was demonstrated in the tibias of mice inoculated with breast cancer MDA-MB-231 bone metastatic cells. This bone loss was prevented by p.o. administration of UBS109 (50 and 150 mg/kg body weight) and i.p. treatment of UBS109 (10 and 20 mg/kg) in vivo. Culture of bone marrow cells obtained from the bone tissues of mice with breast cancer cell bone metastasis showed suppressed osteoblastic mineralization and stimulated osteoclastogenesis ex vivo. These changes were not seen after culture of the bone marrow cells obtained from mice treated with UBS109. Moreover, UBS109 was found to stimulate osteoblastic mineralization and suppress lipopolysaccharide (LPS)-induced osteoclastogenesis in bone marrow cells obtained from normal nude mice in vitro. These findings suggest that the novel curcumin analogue UBS109 prevents breast cancer cell bone metastasis-induced bone loss by stimulating osteoblastic mineralization and suppressing osteoclastogenesis. PMID:24723227

Yamaguchi, Masayoshi; Zhu, Shijun; Zhang, Shumin; Wu, Daqing; Moore, Terry M; Snyder, James P; Shoji, Mamoru

2014-07-01

41

Isolation of primary myofibroblasts from mouse and human colon tissue.  

PubMed

The myofibroblast is a stromal cell of the gastrointestinal (GI) tract that has been gaining considerable attention for its critical role in many GI functions. While several myofibroblast cell lines are commercially available to study these cells in vitro, research results from a cell line exposed to experimental cell culture conditions have inherent limitations due to the overly reductionist nature of the work. Use of primary myofibroblasts offers a great advantage in terms of confirming experimental findings identified in a cell line. Isolation of primary myofibroblasts from an animal model allows for the study of myofibroblasts under conditions that more closely mimic the disease state being studied. Isolation of primary myofibroblasts from human colon tissue provides arguably the most relevant experimental data, since the cells come directly from patients with the underlying disease. We describe a well-established technique that can be utilized to isolate primary myofibroblasts from both mouse and human colon tissue. These isolated cells have been characterized to be alpha-smooth muscle actin and vimentin-positive, and desmin-negative, consistent with subepithelial intestinal myofibroblasts. Primary myofibroblast cells can be grown in cell culture and used for experimental purposes over a limited number of passages. PMID:24145735

Khalil, Hassan; Nie, Wenxian; Edwards, Robert A; Yoo, James

2013-01-01

42

Bone and mineral metabolism in patients with primary aldosteronism.  

PubMed

Primary aldosteronism represents major cause of secondary hypertension, strongly associated with high cardiovascular morbidity and mortality. Aldosterone excess may influence mineral homeostasis, through higher urinary calcium excretion inducing secondary increase of parathyroid hormone. Recently, in a cohort of PA patients a significant increase of primary hyperparathyroidism was found, suggesting a bidirectional functional link between the adrenal and parathyroid glands. The aim of this study was to evaluate the impact of aldosterone excess on mineral metabolism and bone mass density. In 73 PA patients we evaluated anthropometric and biochemical parameters, renin-angiotensin-aldosterone system, calcium-phosphorus metabolism, and bone mineral density; control groups were 73 essential hypertension (EH) subjects and 40 healthy subjects. Compared to HS and EH, PA subjects had significantly lower serum calcium levels and higher urinary calcium excretion. Moreover, PA patients showed higher plasma PTH, lower serum 25(OH)-vitamin D levels, higher prevalence of vitamin D deficiency (65% versus 25% and 25%; P < 0.001), and higher prevalence of osteopenia/osteoporosis (38.5 and 10.5%) than EH (28% and 4%) and NS (25% and 5%), respectively. This study supports the hypothesis that bone loss and fracture risk in PA patients are potentially the result of aldosterone mediated hypercalciuria and the consecutive secondary hyperparathyroidism. PMID:24864141

Petramala, Luigi; Zinnamosca, Laura; Settevendemmie, Amina; Marinelli, Cristiano; Nardi, Matteo; Concistrè, Antonio; Corpaci, Francesco; Tonnarini, Gianfranco; De Toma, Giorgio; Letizia, Claudio

2014-01-01

43

Bone and Mineral Metabolism in Patients with Primary Aldosteronism  

PubMed Central

Primary aldosteronism represents major cause of secondary hypertension, strongly associated with high cardiovascular morbidity and mortality. Aldosterone excess may influence mineral homeostasis, through higher urinary calcium excretion inducing secondary increase of parathyroid hormone. Recently, in a cohort of PA patients a significant increase of primary hyperparathyroidism was found, suggesting a bidirectional functional link between the adrenal and parathyroid glands. The aim of this study was to evaluate the impact of aldosterone excess on mineral metabolism and bone mass density. In 73 PA patients we evaluated anthropometric and biochemical parameters, renin-angiotensin-aldosterone system, calcium-phosphorus metabolism, and bone mineral density; control groups were 73 essential hypertension (EH) subjects and 40 healthy subjects. Compared to HS and EH, PA subjects had significantly lower serum calcium levels and higher urinary calcium excretion. Moreover, PA patients showed higher plasma PTH, lower serum 25(OH)-vitamin D levels, higher prevalence of vitamin D deficiency (65% versus 25% and 25%; P < 0.001), and higher prevalence of osteopenia/osteoporosis (38.5 and 10.5%) than EH (28% and 4%) and NS (25% and 5%), respectively. This study supports the hypothesis that bone loss and fracture risk in PA patients are potentially the result of aldosterone mediated hypercalciuria and the consecutive secondary hyperparathyroidism.

Petramala, Luigi; Zinnamosca, Laura; Settevendemmie, Amina; Marinelli, Cristiano; Nardi, Matteo; Concistre, Antonio; Corpaci, Francesco; Tonnarini, Gianfranco; De Toma, Giorgio; Letizia, Claudio

2014-01-01

44

Mechanical loading, damping, and load-driven bone formation in mouse tibiae  

PubMed Central

Mechanical loads play a pivotal role in the growth and maintenance of bone and joints. Although loading can activate anabolic genes and induce bone remodeling, damping is essential for preventing traumatic bone injury and fracture. In this study we investigated the damping capacity of bone, joint tissue, muscle, and skin using a mouse hindlimb model of enhanced loading in conjunction with finite element modeling to model bone curvature. Our hypothesis was that loads were primarily absorbed by the joints and muscle tissue, but that bone also contributed to damping through its compression and natural bending. To test this hypothesis, fresh mouse distal lower limb segments were cyclically loaded in axial compression in sequential bouts, with each subsequent bout having less surrounding tissue. A finite element model was generated to model effects of bone curvature in silico. Two damping-related parameters (phase shift angle and energy loss) were determined from the output of the loading experiments. Interestingly, the experimental results revealed that the knee joint contributed to the largest portion of the damping capacity of the limb, and bone itself accounted for approximately 38% of the total phase shift angle. Computational results showed that normal bone curvature enhanced the damping capacity of the bone by approximately 40%, and the damping effect grew at an accelerated pace as curvature was increased. Although structural curvature reduces critical loads for buckling in beam theory, evolution apparently favors maintaining curvature in the tibia. Histomorphometric analysis of the tibia revealed that in response to axial loading, bone formation was significantly enhanced in the regions that were predicted to receive a curvature-induced bending moment. These results suggest that in addition to bone’s compressive damping capacity, surrounding tissues, as well as naturally-occurring bone curvature, also contribute to mechanical damping, which may ultimately affect bone remodeling and bone quality.

Dodge, Todd; Wanis, Mina; Ayoub, Ramez; Zhao, Liming; Watts, Nelson B.; Bhattacharya, Amit; Akkus, Ozan; Robling, Alexander; Yokota, Hiroki

2012-01-01

45

Resveratrol Inhibited Hydroquinone-Induced Cytotoxicity in Mouse Primary Hepatocytes  

PubMed Central

Hydroquinone (1,4-benzenediol) has been widely used in clinical situations and the cosmetic industry because of its depigmenting effects. Most skin-lightening hydroquinone creams contain 4%–5% hydroquinone. We have investigated the role of resveratrol in prevention of hydroquinone induced cytotoxicity in mouse primary hepatocytes. We found that 400 µM hydroquinone exposure alone induced apoptosis of the cells and also resulted in a significant drop of cell viability compared with the control, and pretreatment of resveratrol to a final concentration of 0.5 mM 1 h before hydroquinone exposure did not show a significant improvement in the survival rate of the hepatocytes, however, relatively higher concentrations of resveratrol (?1 mM) inhibited apoptosis of the mouse primary hepatocytes and increased cell viability in a dose-dependent manner, and in particular the survival rate of the hepatocytes was recovered from 28% to near 100% by 5 mM resveratrol. Interestingly, pretreatment with resveratrol for longer time (24 h), even in very low concentrations (50 µM, 100 µM), blocked the damage of hydroquinone to the cells. We also observed that resveratrol pretreatment suppressed hydroquinone-induced expression of cytochrome P450 2E1 mRNA dose-dependently. The present study suggests that resveratrol protected the cells against hydroquinone-induced toxicity through its antioxidant function and possibly suppressive effect on the expression of cytochrome P450 2E1.

Wang, Da-Hong; Ootsuki, Yoshie; Fujita, Hirofumi; Miyazaki, Masahiro; Yie, Qinxia; Tsutsui, Ken; Sano, Kuniaki; Masuoka, Noriyoshi; Ogino, Keiki

2012-01-01

46

[Treatment of primary bone sarcoma of the jaws].  

PubMed

Head and neck bone sarcomas are very uncommon tumors. Therefore, treatment modalities are not clearly established. The medical record of 12 patients with bone sarcoma of the jaw were reviewed. Six patients with osteosarcoma underwent primary chemotherapy followed by wide surgical resection, radiation therapy or combined radiosurgical treatment. The 2 and 5 years survivals were 66% and 40% respectively. Five patients with chondrosarcoma were treated by wide surgical resection alone or combined with postoperative radiotherapy and possibly chemotherapy. All patients were alive; the mean follow-up was 9 years. One patient had Ewing's tumor. Osteosarcoma and chondrosarcoma in head and neck patients have a high rate of local recurrences. Surgery is the mainstay of treatment. Patients with voluminous tumors and high-grade lesions should receive postoperative radiotherapy. The role of chemotherapy has not been defined. PMID:7939361

Barthélémy, I; Coustal, B; Michelet, V; Pinsolle, J; Siberchicot, F; Caix, P; Michelet, F X

1994-01-01

47

Primary homologies of the circumorbital bones of snakes.  

PubMed

Some snakes have two circumorbital ossifications that in the current literature are usually referred to as the postorbital and supraorbital. We review the arguments that have been proposed to justify this interpretation and provide counter-arguments that reject those conjectures of primary homology based on the observation of 32 species of lizards and 81 species of snakes (both extant and fossil). We present similarity arguments, both topological and structural, for reinterpretation of the primary homologies of the dorsal and posterior orbital ossifications of snakes. Applying the test of similarity, we conclude that the posterior orbital ossification of snakes is topologically consistent as the homolog of the lacertilian jugal, and that the dorsal orbital ossification present in some snakes (e.g., pythons, Loxocemus, and Calabaria) is the homolog of the lacertilian postfrontal. We therefore propose that the terms postorbital and supraorbital should be abandoned as reference language for the circumorbital bones of snakes, and be replaced with the terms jugal and postfrontal, respectively. The primary homology claim for the snake "postorbital" fails the test of similarity, while the term "supraorbital" is an unnecessary and inaccurate application of the concept of a neomorphic ossification, for an element that passes the test of similarity as a postfrontal. This reinterpretation of the circumorbital bones of snakes is bound to have important repercussions for future phylogenetic analyses and consequently for our understanding of the origin and evolution of snakes. PMID:23630161

Palci, Alessandro; Caldwell, Michael W

2013-09-01

48

Mechanical loading, damping, and load-driven bone formation in mouse tibiae.  

PubMed

Mechanical loads play a pivotal role in the growth and maintenance of bone and joints. Although loading can activate anabolic genes and induce bone remodeling, damping is essential for preventing traumatic bone injury and fracture. In this study we investigated the damping capacity of bone, joint tissue, muscle, and skin using a mouse hindlimb model of enhanced loading in conjunction with finite element modeling to model bone curvature. Our hypothesis was that loads were primarily absorbed by the joints and muscle tissue, but that bone also contributed to damping through its compression and natural bending. To test this hypothesis, fresh mouse distal lower limb segments were cyclically loaded in axial compression in sequential bouts, with each subsequent bout having less surrounding tissue. A finite element model was generated to model effects of bone curvature in silico. Two damping-related parameters (phase shift angle and energy loss) were determined from the output of the loading experiments. Interestingly, the experimental results revealed that the knee joint contributed to the largest portion of the damping capacity of the limb, and bone itself accounted for approximately 38% of the total phase shift angle. Computational results showed that normal bone curvature enhanced the damping capacity of the bone by approximately 40%, and the damping effect grew at an accelerated pace as curvature was increased. Although structural curvature reduces critical loads for buckling in beam theory, evolution apparently favors maintaining curvature in the tibia. Histomorphometric analysis of the tibia revealed that in response to axial loading, bone formation was significantly enhanced in the regions that were predicted to receive a curvature-induced bending moment. These results suggest that in addition to bone's compressive damping capacity, surrounding tissues, as well as naturally-occurring bone curvature, also contribute to mechanical damping, which may ultimately affect bone remodeling and bone quality. PMID:22878153

Dodge, Todd; Wanis, Mina; Ayoub, Ramez; Zhao, Liming; Watts, Nelson B; Bhattacharya, Amit; Akkus, Ozan; Robling, Alexander; Yokota, Hiroki

2012-10-01

49

Probiotic L. reuteri Treatment Prevents Bone Loss in a Menopausal Ovariectomized Mouse Model.  

PubMed

Estrogen deficiency is a major risk factor for osteoporosis that is associated with bone inflammation and resorption. Half of women over the age of 50 will experience an osteoporosis related fracture in their lifetime, thus novel therapies are needed to combat post-menopausal bone loss. Recent studies suggest an important role for gut-bone signaling pathways and the microbiota in regulating bone health. Given that the bacterium Lactobacillus reuteri ATCC PTA 6475 (L. reuteri) secretes beneficial immunomodulatory factors, we examined if this candidate probiotic could reduce bone loss associated with estrogen deficiency in an ovariectomized (Ovx) mouse menopausal model. Strikingly, L. reuteri treatment significantly protected Ovx mice from bone loss. Osteoclast bone resorption markers and activators (Trap5 and RANKL) as well as osteoclastogenesis are significantly decreased in L. reuteri-treated mice. Consistent with this, L. reuteri suppressed Ovx-induced increases in bone marrow CD4+ T-lymphocytes (which promote osteoclastogenesis) and directly suppressed osteoclastogenesis in vitro. We also identified that L. reuteri treatment modifies microbial communities in the Ovx mouse gut. Together, our studies demonstrate that L. reuteri treatment suppresses bone resorption and loss associated with estrogen deficiency. Thus, L. reuteri treatment may be a straightforward and cost-effective approach to reduce post-menopausal bone loss. J. Cell. Physiol. 229: 1822-1830, 2014. © 2014 Wiley Periodicals, Inc. PMID:24677054

Britton, Robert A; Irwin, Regina; Quach, Darin; Schaefer, Laura; Zhang, Jing; Lee, Taehyung; Parameswaran, Narayanan; McCabe, Laura R

2014-11-01

50

Primary calvarial bone grafting in oro-ocular clefts.  

PubMed

Oro-ocular clefts were classified by Tessier as 3-11 or 4-10 facial clefts. These rare clefts require both bony and soft tissue correction and present a significant challenge to the innovative skill of the craniofacial surgeon. A review of three male children with oro-ocular clefts and their management (including surgical planning and operative treatment) is presented. We would propose that the use of calvarial bone grafting as part of the primary closure is regarded as desirable based on our experience. PMID:19464971

O'Sullivan, J B; Earley, M J

2010-05-01

51

Evaluation of critical size defects of mouse calvarial bone: An organ culture study.  

PubMed

Mouse calvarial organ culture has been used widely for the study of bone biology. The purpose of this study was to evaluate the healing potential of neonatal mouse parietal defects in different culture media. The critical size defect (CSD) was also investigated. The parietal bones of neonatal mice were used. Full-thickness, 0.8-mm circular defects were created through the bones from one litter of mice. The bones were divided into three groups: Dulbecco's Modified Eagle Medium (DMEM) group, DMEM/osteogenic medium (OM) group, and OM group. Cultures were analyzed with microcomputed-tomography, dissecting-microscope, phase-contrast-microscope, Von Kossa stain, scanning-electron-microscopy, and energy-dispersive-X-ray. Continuous bone formation of parietal bones was observed in all groups. Defects in the DMEM/OM group showed the highest healing potential and exhibited woven bone formation. Defects in the OM group showed limited bone healing at the defect edge. Defects in the DMEM group showed fibrous healing. The most effective culture medium (DMEM/OM) was used to determine the CSD of mouse calvaria in a separate experiment. Circular defects (diameters: 0.8, 1.0, and 1.5 mm) were made in the parietal bones from another litter of mice. The bones were analyzed with microcomputed-tomography, and phase-contrast-microscopy. The bone filling percentages of different size defects were statistically significant: 1.5-mm defects (4.49%), 1.0-mm defects (47.65%), and 0.8-mm defects (73.45%). In three culture conditions, DMEM/OM was the most effective approach to repair bone defects. A 1.5 mm in diameter, full-thickness parietal defect was found to be the CSD under the DMEM/OM culture conditions. PMID:19937748

Wu, Xiaohong; Downes, Sandra; Watts, David C

2010-05-01

52

Application of retinoic acid to obtain osteocytes cultures from primary mouse osteoblasts.  

PubMed

The need for osteocyte cultures is well known to the community of bone researchers; isolation of primary osteocytes is difficult and produces low cell numbers. Therefore, the most widely used cellular system is the osteocyte-like MLO-Y4 cell line. The method here described refers to the use of retinoic acid to generate a homogeneous population of ramified cells with morphological and molecular osteocyte features. After isolation of osteoblasts from mouse calvaria, all-trans retinoic acid (ATRA) is added to cell medium, and cell monitoring is conducted daily under an inverted microscope. First morphological changes are detectable after 2 days of treatment and differentiation is generally complete in 5 days, with progressive development of dendrites, loss of the ability to produce extracellular matrix, down-regulation of osteoblast markers and up-regulation of osteocyte-specific molecules. Daily cell monitoring is needed because of the inherent variability of primary cells, and the protocol can be adapted with minimal variation to cells obtained from different mouse strains and applied to transgenic models. The method is easy to perform and does not require special instrumentation, it is highly reproducible, and rapidly generates a mature osteocyte population in complete absence of extracellular matrix, allowing the use of these cells for unlimited biological applications. PMID:24894124

Mattinzoli, Deborah; Messa, Piergiorgio; Corbelli, Alessandro; Ikehata, Masami; Mondini, Anna; Zennaro, Cristina; Armelloni, Silvia; Li, Min; Giardino, Laura; Rastaldi, Maria Pia

2014-01-01

53

Fine-needle aspirate of primary lymphoma of bone.  

PubMed

Primary lymphoma of bone (PLB) is a rare bone tumor. Fine-needle aspirates (FNA) were done on large destructive bone tumors from 2 elderly men, and both were initially read as inconclusive for malignancy because of scant cellularity. On retrospective study of the FNA slides after examining tissue histology, low numbers of diagnostic cells for lymphoma were recognized on the smears. There was extensive crush artifact, and most intact cells were stripped of their cytoplasm. In neither case was enough material harvested to make cell blocks or to perform special studies. Tissue histology disclosed abundant fibroconnective tissue stroma which probably made it difficult to acquire adequate FNA specimens. Another FNA done on a postoperative PLB tissue specimen disclosed similar features. Our experience in the 3 cases is consistent with the view that even though smears show scant cellularity, the diagnosis of PLB can at least be suggested by FNA. It is therefore important not to undercall the specimen because of low cellularity, and recommendation for tissue diagnosis can be given. This process is facilitated by a high index of suspicion based on clinical and radiographic findings. PMID:8989547

Htwe, W M; Lucas, D R; Bedrossian, C W; Ryan, J R

1996-12-01

54

Endochondral bone formation in embryonic mouse pre-metatarsals  

NASA Technical Reports Server (NTRS)

Long term exposure to a reduced gravitational environment has a deleterious effect on bone. The developmental events which occur prior to initial bone deposition will provide insight into the regulation of mature bone physiology. We have characterized a system in which the events preceding bone formation take place in an isolated in vitro organ culture environment. We show that cultured pre-metatarsal tissue parallels development of pre-metatarsal tissue in the embryo. Both undergo mesenchyme differentiation and morphogenesis to form a cartilage rod, which resembles the future bone, followed by terminal chondrocyte differentiation in a definite morphogenetic pattern. These sequential steps occur prior to osteoblast maturation and bone matrix deposition in the developing organism. Alkaline phosphatase (ALP) activity is a distinctive enzymatic marker for mineralizing tissues. We have measured this activity throughout pre-metatarsal development and show (a) where in the tissue it is predominantly found, and (b) that this is indeed the mineralizing isoform of the enzyme.

Klement, B. J.; Spooner, B. S.

1992-01-01

55

Cadmium stimulates osteoclast-like multinucleated cell formation in mouse bone marrow cell cultures  

SciTech Connect

Most of cadmium (Cd)-treated animals have been reported to show osteoporosis-like changes in bones. This suggests that Cd may promote bone loss by a direct action on bone. It was found that Cd stimulated prostaglandin E{sub 2}(PGE{sub 2}) production in the osteoblast-like cell, MC3T3-E1. Therefore, Cd stimulates bone resorption by increasing PGE{sub 2} production. Recently, several bone marrow cell culture systems have been developed for examining the formation of osteoclast-like multinucleated cells in vitro. As osteoblasts produce PGE{sub 2} by Cd-induced cyclooxygenase and may play an important role in osteoclast formation, the present study was undertaken to clarify the possibility that Cd might stimulate osteoclast formation in a mouse bone marrow culture system.

Miyahara, Tatsuro; Takata, Masakazu; Miyata, Masaki; Nagai, Miyuki; Sugure, Akemi; Kozuka, Hiroshi; Kuze, Shougo (Toyama Medical and Pharmaceutical Univ. (Japan))

1991-08-01

56

Mineral metabolism in isolated mouse long bones: Opposite effects of microgravity on mineralization and resorption  

NASA Technical Reports Server (NTRS)

An experiment using isolated skeletal tissues under microgravity, is reported. Fetal mouse long bones (metatarsals) were cultured for 4 days in the Biorack facility of Spacelab during the IML-1 (International Microgravity Laboratory) mission of the Space Shuttle. Overall growth was not affected, however glucose consumption was significantly reduced under microgravity. Mineralization of the diaphysis was also strongly reduced under microgravity as compared to the on-board 1 g group. In contrast, mineral resorption by osteoclasts was signficantly increased. These results indicate that these fetal mouse long bones are a sensitive and useful model to further study the cellular mechanisms involved in the changed mineral metabolism of skeletal tissues under microgravity.

Veldhuijzen, Jean Paul; Vanloon, Jack J. W. A.

1994-01-01

57

Tumor-induced injury of primary afferent sensory nerve fibers in bone cancer pain  

Microsoft Academic Search

Bone is the most common site of chronic pain in patients with metastatic cancer. What remains unclear are the mechanisms that generate this pain and why bone cancer pain can be so severe and refractory to treatment with opioids. Here we show that following injection and confinement of NCTC 2472 osteolytic tumor cells within the mouse femur, tumor cells sensitize

Christopher M. Peters; Joseph R. Ghilardi; Cathy P. Keyser; Kazufumi Kubota; Theodore H. Lindsay; Nancy M. Luger; David B. Mach; Matthew J. Schwei; Molly A. Sevcik; Patrick W. Mantyh

2005-01-01

58

Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Neurocytes and Adipocytes Induced in vitro  

Microsoft Academic Search

Objective) To establish a system for isolation and culture of bone marrow mesenchymal stem cells (MSCs) from Kun-ming mouse in intro and to identify the characteristics of the cells, directional neuronal and adipocytic induction after culture expansion. (Method)MSCs were isolated and cultivated from the bone marrow of Kun-ming mice by investigating their adherence-dependent growth characters. The morphological characters of MSCs

ZHAO Xing; BAI Xue-jin; FENG Ji-wu; DONG Ya-juan

2008-01-01

59

Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Neurocytes and Adipocytes Induced in vitro  

Microsoft Academic Search

Objective) To establish a system for isolation and culture of bone marrow mesenchymal stem cells (MSCs) from Kun-ming mouse in intro and to identify the characteristics of the cells, directional neuronal and adipocytic induction after culture expansion. (Method)MSCs were isolated and cultivated from the bone marrow of Kun-ming mice by investigating their adherence-dependent growth characters. The morphological characters of MSCs

60

Primary myoepithelioma of bone: a report of 8 cases.  

PubMed

The clinical and pathologic features of 8 primary myoepitheliomas of bone were analyzed. There were 5 female and 3 male patients who ranged in age from 16 to 49 (mean, 33.5) years. Three tumors arose in the ilium, 2 in the tibia, and 1 each in the maxilla, sacrum, and L1 vertebral body. Microscopically, the tumors had a solid, lobulated, reticular, or storiform growth pattern and were predominantly composed of spindle-shaped cells arranged in intersecting fascicles with eosinophilic cytoplasm. The round to polygonal epithelioid cells were arranged randomly or formed small clusters and contained variable amounts of eosinophilic or clear cytoplasm. Immunohistochemically, all the tumors were positive for vimentin and S100 protein, and 7 were positive for epithelial membrane antigen. No tumors were positive for keratin (AE1.3/CAM5.2). Smooth muscle actin was positive in 3 tumors and negative in 4, whereas desmin was negative in all 7 tumors tested. Nuclear staining for p63 was negative in 3 tested tumors. Staining for GFAP and CD34 was performed on 4 and 5 tumors, respectively, and all showed no expression. Fluorescence in situ hybridization for EWSR1 rearrangement was performed in 7 tumors. Five tumors (71%) showed the presence of EWSR1 gene rearrangement, and 2 were negative. Cytogenetic studies conducted on 1 tumor showed 46,XY,t(1;22)(q21;q12) associated with EWSR1-PBX1 fusion. Surgical procedures included curettage in 3 patients, resection in 3 patients, and 2 patients only had an open biopsy. Follow-up information was available for 4 patients; all remain free of disease with no recurrence. Although experience with primary myoepithelioma of bone is limited, histologically, banal tumors appear to behave in a benign manner, and conservative surgery appears to be sufficient treatment. Immunohistochemical and molecular analyses are helpful in their accurate identification. PMID:23681076

Kurzawa, Pawel; Kattapuram, Susan; Hornicek, Francis J; Antonescu, Cristina R; Rosenberg, Andrew E; Nielsen, G Petur

2013-07-01

61

Stromal Gene Expression and Function in Primary Breast Tumors that Metastasize to Bone Cancer.  

National Technical Information Service (NTIS)

A clinically relevant syngeneic model of breast cancer metastasis has been used to determine gene expression alterations that occur both between primary breast cancers with varying metastatic potential and between matched primary and bone metastases. We h...

B. Parker R. L. Anderson

2004-01-01

62

Morphometric studies on mouse bone using a computer-based image-analysis system.  

PubMed

The morphological structure of the ilium, femur, third lumbar vertebra and a central caudal vertebra of the female CBA mouse has been studied using 5 micrometer thick, plastic embedded, transverse and longitudinal sections. The sections were analysed on a Quantimet 720, system 30, image analyser connected on-line to a PDP11 computer. Separate endosteal and periosteal surface to volume ratios were calculated for each position of sampling in each bone. For this calculation the anisotrophy of the bone was estimated from measurements of mean chord lengths in longitudinal sections of the bone using a new analytical technique. Chord length distributions in transverse sections of bone were also measured and the relevance of such measurements to the study of morphological changes in the bone and its included marrow are briefly discussed. PMID:7012370

Green, D; Howells, G R; Thorne, M C

1981-04-01

63

Effects of 810 nm laser on mouse primary cortical neurons  

NASA Astrophysics Data System (ADS)

In the past four decades numerous studies have reported the efficacy of low level light (laser) therapy (LLLT) as a treatment for diverse diseases and injuries. Recent studies have shown that LLLT can biomodulate processes in the central nervous system and has been extensively studied as a stroke treatment. However there is still a lack of knowledge on the effects of LLLT at the cellular level in neurons. The present study aimed to study the effect of 810 nm laser on several cellular processes in primary cortical neurons cultured from mouse embryonic brains. Neurons were irradiated with light dose of 0.03, 0.3, 3, 10 and 30 J/cm2 and intracellular levels of reactive oxygen species, nitric oxide and calcium were measured. The changes in mitochondrial function in response to light were studied in terms of adenosine triphosphate (ATP) and mitochondrial membrane potential (MMP). Light induced a significant increase in calcium, ATP and MMP at lower fluences and a decrease at higher fluence. ROS was induced significantly by light at all light doses. Nitric oxide levels also showed an increase on treatment with light. The results of the present study suggest that LLLT at lower fluences is capable of inducing mediators of cell signaling process which in turn may be responsible for the biomodulatory effects of the low level laser. At higher fluences beneficial mediators are reduced but potentially harmful mediators are increased thus offering an explanation for the biphasic dose response.

Kharkwal, Gitika B.; Sharma, Sulbha K.; Huang, Ying-Ying; de Taboada, Luis; McCarthy, Thomas; Hamblin, Michael R.

2011-02-01

64

Prospective isolation of mesenchymal stem cells from mouse compact bone.  

PubMed

Bone marrow from numerous species, including rodents and man, has been shown to contain a rare population of cells known as marrow stromal cells or mesenchymal stem cells (MSC). Given the innate ability of these cells to give rise to multiple tissue types including bone, fat and cartilage, there is considerable interest in utilizing MSC in a broad repertoire of cell-based therapies for the treatment of human disease. In order for such therapies to be realized, a preclinical animal model in which to refine strategies utilizing MSC is required.We have described methodology allowing for the prospective isolation by fluorescence activated cell sorting (FACS) of a highly purified population of MSC from murine compact bone (CB). These cells are multipotent and capable of extensive proliferation in vitro and thus represent an ideal source of cells with which to explore both the fundamental biology of MSC and their efficacy in a variety of cellular therapies. PMID:19089361

Short, Brenton J; Brouard, Nathalie; Simmons, Paul J

2009-01-01

65

Primary malignant tumours of bone and soft tissue in the elderly  

Microsoft Academic Search

Aims. This study reports outcome, functional results and quality of life of 45 elderly patients with age over 70 after surgery for primary malignant bone and soft tissue tumours.Methods. There were 24 primary malignant bone tumours and 21 soft tissue sarcomas. The most frequent diagnoses were: chondrosarcoma, malignant fibrous histiocytoma and liposarcoma. Local tumour resection with and without osteosynthesis, endoprostheses,

M. Buchner; L. Bernd; A. Zahlten-Hinguranage; D. Sabo

2004-01-01

66

Effect of sex hormones on bone in primary osteoporosis  

PubMed Central

The effect of sex hormones on bone tissue was studied in 12 osteoporotic patients. Surfaces of bone undergoing formation and resorption were determined by quantitative microradiography of iliac crest biopsy samples before and after treatment with estrogens in 11 postmenopausal women and with testosterone in one gonadally competent man. Before treatment, bone resorption was greater than normal in all but one patient and bone formation was normal. After treatment, bone resorption decreased to within the normal range in all patients, and bone formation did not change significantly. Biochemical studies showed significant decreases in serum calcium, phosphorus, and alkaline phosphatase levels and in urinary excretion of calcium and hydroxyproline. These changes are believed to be the consequence of the effect of the hormones on bone. The data indicate that the major effect of sex hormones in osteoporosis is an inhibition of bone resorption. Images

Riggs, B. Lawrence; Jowsey, Jenifer; Kelly, Patrick J.; Jones, James D.; Maher, Frank T.

1969-01-01

67

Efficacy of serotonin inhibition in mouse models of bone loss.  

PubMed

In a proof-of-concept study it was shown that decreasing synthesis of gut serotonin through a small molecule inhibitor of Tph1 could prevent and treat ovariectomy-induced osteoporosis in young mice and rats. In this study, we define the minimal efficacy of this Tph1 inhibitor, demonstrate that its activity is improved with the duration of treatment, and show that its anabolic effect persists on interruption. Importantly, given the prevalence of osteoporosis in the aging population, we then show that Tph1 inhibition rescues ovariectomy-induced bone loss in aged mice. It also cures the low bone mass of Lrp5-deficient mice through a sole anabolic effect. Lastly, we provide evidence that inhibition of gut serotonin synthesis can work in concert with an antiresorptive agent to increase bone mass in ovariectomized mice. This study provides a more comprehensive view of the anabolic efficacy of Tph1 inhibitors and further establishes the spectrum of their therapeutic potential in the treatment of bone-loss disorders. PMID:21608033

Inose, Hiroyuki; Zhou, Bin; Yadav, Vijay K; Guo, X Edward; Karsenty, Gerard; Ducy, Patricia

2011-09-01

68

Comparative analyses of B cell populations in trout kidney and mouse bone marrow; establishing "B cell signatures"  

PubMed Central

This study aimed to identify the frequency and distribution of developing B cell populations in the kidney of the rainbow trout, using four molecular B cell markers that are highly conserved between species, including two transcription factors, Pax5 and EBF1, recombination activating gene RAG1, and the immunoglobulin heavy chain mu. Three distinct B cell stages were defined: early developing B cells (CLP, pro-B, and early pre-B cells), late developing B cell (late pre-B, immature B, and mature B cells), and IgM-secreting cells. Developmental stage-specific, combinatorial expression of Pax5, EBF1, RAG1 and immunoglobulin mu was determined in trout anterior kidney cells by flow cytometry. Trout staining patterns were compared to a well-defined primary immune tissue, mouse bone marrow, and using mouse surface markers B220 and CD43. A remarkable level of similarity was uncovered between the primary immune tissues of both species. Subsequent analysis of the entire trout kidney, divided into five contiguous segments K1-K5, revealed a complex pattern of early developing, late developing, and IgM-secreting B cells. Patterns in anterior kidney segment K1 were most similar to those of mouse bone marrow, while the most posterior part of the kidney, K5, had many IgM-secreting cells, but lacked early developing B cells. A potential second B lymphopoiesis site was uncovered in segment K4 of the kidney. The B cell patterns, or “B cell signatures” described here provide information on the relative abundance of distinct developing B cell populations in the trout kidney, and can be used in future studies on B cell development in other vertebrate species.

Zwollo, Patty; Mott, Katrina; Barr, Maggie

2010-01-01

69

Marrow Stromal Cell-Based Cyclooxygenase 2 Ex Vivo Gene-Transfer Strategy Surprisingly Lacks Bone-Regeneration Effects and Suppresses the Bone-Regeneration Action of Bone Morphogenetic Protein 4 in a Mouse Critical-Sized Calvarial Defect Model  

Microsoft Academic Search

This study evaluated whether the murine leukemia virus (MLV)–based cyclooxygenase-2 (Cox-2) ex vivo gene-transfer strategy promotes healing of calvarial defects and\\/or synergistically enhances bone morphogenetic\\u000a protein (BMP) 4–mediated bone regeneration. Gelatin scaffolds impregnated with mouse marrow stromal cells (MSCs) transduced\\u000a with MLV-expressing BMP4, Cox-2, or a control gene were implanted into mouse calvarial defects. Bone regeneration was assessed by X-ray,

K.-H. William Lau; Reinhard Gysin; Shin-Tai Chen; Jon E. Wergedal; David J. Baylink; Subburaman Mohan

2009-01-01

70

Interleukin-10 inhibits the osteogenic activity of mouse bone marrow.  

PubMed

Murine bone marrow cells synthesize bone proteins, including alkaline phosphatase (ALP), collagen type I, and osteocalcin, and form a mineralized extracellular matrix when cultured in the presence of beta-glycerophosphate and vitamin C. Interleukin-10 (IL-10) suppressed the synthesis of these bone proteins and mineralization without affecting cell proliferation. In addition, mRNA levels for the latter proteins were reduced in IL-10-treated cultures. This inhibitory effect was most outspoken when IL-10 was added before ALP activity peaked, eg, day 15 of culture. No significant effect was observed when IL-10 was added at later time points. This finding suggests that IL-10 acts at osteogenic differentiation stages that precede ALP expression but is ineffective on cells that progressed beyond this maturation stage. Likewise, IL-10 appeared to be unable to block both ALP activity and collagen synthesis in the preosteosteoblastic cell lines MN7 and MC3T3 that constitutively synthesize these proteins. Whereas IL-10 did not alter the number of fibroblast colony-forming cells of the marrow, it significantly reduced their osteogenic differentiation potential. In contrast to control cultures, IL-10-treated stroma was unable to either synthesize osteocalcin or to mineralize when subcultured over a 25-day period in the absence of IL-10. The inhibitory activity of IL-10 coincided with significant changes in stroma morphology. Whereas control cultures contained mainly flat adherent polygonal cells, significant numbers of rounded semiadherent to nonadherent cells were observed in the presence of IL-10. Scanning and transmission electron microscopy showed that, in contrast to control cultures, IL-10-treated stromas completely lacked a mineralized extracellular matrix. Collectively, these data suggest that IL-10 may have important regulatory effects on bone biology because of its capacity to downregulate early steps of osteogenic differentiation. PMID:8400287

Van Vlasselaer, P; Borremans, B; Van Den Heuvel, R; Van Gorp, U; de Waal Malefyt, R

1993-10-15

71

Effect of cyclophosphamide and electromagnetic fields on mouse bone marrow  

Microsoft Academic Search

The authors have previously shown that the exposure to low frequency pulsing electromagnetic fields (PEMF) of mice X-ray irradiated resulted in an increased damage to the bone marrow. The series of experiments here reported were designed to investigate the effect of PEMF exposure after intraperitoneum injection of 200mg\\/kg of cyclophosphamide (CY). Control mice were CY injected only; experimental mice were

R. Cadossi; P. Zucchini; G. Emilia; G. Torelli

1990-01-01

72

Effect of cyclophosphamide and electromagnetic fields on mouse bone marrow  

SciTech Connect

The authors have previously shown that the exposure to low frequency pulsing electromagnetic fields (PEMF) of mice X-ray irradiated resulted in an increased damage to the bone marrow. The series of experiments here reported were designed to investigate the effect of PEMF exposure after intraperitoneum injection of 200mg/kg of cyclophosphamide (CY). Control mice were CY injected only; experimental mice were CY injected and then exposed to PEMF. Exposure to PEMF (24 hours/day) increased the rate of decline of white blood cells in peripheral blood. Spleen weight was statistically higher among control mice than among mice exposed to PEMF at day 6, 8 and 10 after CY injection. Spleen autoradiography proved to be higher among PEMF exposed mice than among controls at day 8 and 9 after CY injection. The grafting efficiency of the bone marrow obtained from control mice was higher than the grafting efficiency of the bone marrow recovered from mice exposed to PEMF. All these data indicate that the exposure to PEMF increases the cytotoxic effect of CY.

Cadossi, R.; Zucchini, P.; Emilia, G.; Torelli, G. (Univ. di Modena (Italy))

1990-02-26

73

Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism  

PubMed Central

Primary cilia are sensory organelles that translate extracellular chemical and mechanical cues into cellular responses. Bone is an exquisitely mechanosensitive organ, and its homeostasis depends on the ability of bone cells to sense and respond to mechanical stimuli. One such stimulus is dynamic fluid flow, which triggers biochemical and transcriptional changes in bone cells by an unknown mechanism. Here we report that bone cells possess primary cilia that project from the cell surface and deflect during fluid flow and that these primary cilia are required for osteogenic and bone resorptive responses to dynamic fluid flow. We also show that, unlike in kidney cells, primary cilia in bone translate fluid flow into cellular responses in bone cells independently of Ca2+ flux and stretch-activated ion channels. These results suggest that primary cilia might regulate homeostasis in diverse tissues by allowing mechanical signals to alter cellular activity via tissue-specific pathways. Our identification of a mechanism for mechanotransduction in bone could lead to therapeutic approaches for combating bone loss due to osteoporosis and disuse.

Malone, Amanda M. D.; Anderson, Charles T.; Tummala, Padmaja; Kwon, Ronald Y.; Johnston, Tyler R.; Stearns, Tim; Jacobs, Christopher R.

2007-01-01

74

Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism.  

PubMed

Primary cilia are sensory organelles that translate extracellular chemical and mechanical cues into cellular responses. Bone is an exquisitely mechanosensitive organ, and its homeostasis depends on the ability of bone cells to sense and respond to mechanical stimuli. One such stimulus is dynamic fluid flow, which triggers biochemical and transcriptional changes in bone cells by an unknown mechanism. Here we report that bone cells possess primary cilia that project from the cell surface and deflect during fluid flow and that these primary cilia are required for osteogenic and bone resorptive responses to dynamic fluid flow. We also show that, unlike in kidney cells, primary cilia in bone translate fluid flow into cellular responses in bone cells independently of Ca(2+) flux and stretch-activated ion channels. These results suggest that primary cilia might regulate homeostasis in diverse tissues by allowing mechanical signals to alter cellular activity via tissue-specific pathways. Our identification of a mechanism for mechanotransduction in bone could lead to therapeutic approaches for combating bone loss due to osteoporosis and disuse. PMID:17673554

Malone, Amanda M D; Anderson, Charles T; Tummala, Padmaja; Kwon, Ronald Y; Johnston, Tyler R; Stearns, Tim; Jacobs, Christopher R

2007-08-14

75

Bisphosphonate treatment in the oim mouse model alters bone modeling during growth  

PubMed Central

Osetogenesis Imperfecta (OI) is a heritable disease, which results from an abnormal amount or structure of Type I collagen. Bisphosphonates, a class of synthetic antiresorptive drugs used in osteoporosis management, are also used to decrease fracture incidence and improve quality of life in children with OI. In this study we used the oim mouse to test the hypotheses that pamidronate treatment during active growth 1. produces larger, stronger, stiffer long bone diaphyses without altering bone material properties, and 2. negatively impacts longitudinal bone growth. Our results indicate that femoral cross-sectional moment of inertia in the distal metaphysis tended to increase with pamidronate treatment and that the treated bones are thicker and structurally stiffer, but shorter than their control-dose counterpar

Rao, S.H.; Evans, K.D.; Oberbauer, A.M.; Martin, R.B.

2009-01-01

76

Characterization of a calcitonin gene-related peptide (CGRP) receptor on mouse bone marrow cells.  

PubMed

Calcitonin gene-related peptide, (CGRP), a vasoactive neuropeptide, is found throughout the peripheral nervous system, and CGRP receptors are present on mature lymphocytes. The current studies describe a CGRP receptor on isolated mouse bone marrow cells. The affinity, distribution and specificity of CGRP receptors were analyzed using radioligand binding assays. [125I]CGRP binding in mouse bone marrow cells was dependent on cell concentration and was stable from 5 to 60 min at room temperature. The average Kd is 3.29 +/- 1.24 nM and the average receptor density is 2796 +/- 365 sites/cell. Competition binding analysis found rat alpha and beta CGRP to be the most inhibitory, (Ki values 0.899 and 0.711 nM, respectively), followed by human alpha CGRP and the antagonist human CGRP8-37. The neuropeptides human and salmon calcitonin did not inhibit [125I]CGRP binding to bone marrow cells. The presence of CGRP receptors on mouse bone marrow cells provides further evidence for a direct role for CGRP in modulating the function and differentiation of cellular components of the immune and inflammatory systems. PMID:8278635

Mullins, M W; Ciallella, J; Rangnekar, V; McGillis, J P

1993-11-19

77

Ex vivo determination of bone tissue strains for an in vivo mouse tibial loading model  

PubMed Central

Previous studies introduced the digital image correlation (DIC) as a viable technique for measuring bone strain during loading. In this study, we investigated the sensitivity of a DIC system in determining surface strains in a mouse tibia while loaded in compression through the knee joint. Specifically, we examined the effect of speckle distribution, facet size and overlap, initial vertical alignment of the bone into the loading cups, rotation with respect to cameras, and ex vivo loading configurations on the strain contour maps measured with a DIC system. We loaded tibiae of C57BL/6 mice (12 and 18 weeks old male) up to 12 N at 8 N/min. Images of speckles on the bone surface were recorded at 1 N intervals and DIC was used to compute strains. Results showed that speckles must have the correct size and density with respect to the facet size of choice for the strain distribution to be computed and reproducible. Initial alignment of the bone within the loading cups does not influence the strain distribution measured during peak loading, but bones must be placed in front of the camera with the same orientation in order for strains to be comparable. Finally, the ex vivo loading configurations with the tibia attached to the entire mouse, or to the femur and foot, or only to the foot, showed different strain contour maps. This work provides a better understanding of parameters affecting full field strain measurements from DIC in ex vivo murine tibial loading tests.

Carriero, Alessandra; Abela, Lisa; Pitsillides, Andrew A.; Shefelbine, Sandra J.

2014-01-01

78

Ex vivo determination of bone tissue strains for an in vivo mouse tibial loading model.  

PubMed

Previous studies introduced the digital image correlation (DIC) as a viable technique for measuring bone strain during loading. In this study, we investigated the sensitivity of a DIC system in determining surface strains in a mouse tibia while loaded in compression through the knee joint. Specifically, we examined the effect of speckle distribution, facet size and overlap, initial vertical alignment of the bone into the loading cups, rotation with respect to cameras, and ex vivo loading configurations on the strain contour maps measured with a DIC system. We loaded tibiae of C57BL/6 mice (12 and 18 weeks old male) up to 12N at 8N/min. Images of speckles on the bone surface were recorded at 1N intervals and DIC was used to compute strains. Results showed that speckles must have the correct size and density with respect to the facet size of choice for the strain distribution to be computed and reproducible. Initial alignment of the bone within the loading cups does not influence the strain distribution measured during peak loading, but bones must be placed in front of the camera with the same orientation in order for strains to be comparable. Finally, the ex vivo loading configurations with the tibia attached to the entire mouse, or to the femur and foot, or only to the foot, showed different strain contour maps. This work provides a better understanding of parameters affecting full field strain measurements from DIC in ex vivo murine tibial loading tests. PMID:24835472

Carriero, Alessandra; Abela, Lisa; Pitsillides, Andrew A; Shefelbine, Sandra J

2014-07-18

79

Survival of Free and Encapsulated Human and Rat Islet Xenografts Transplanted into the Mouse Bone Marrow  

PubMed Central

Bone marrow was recently proposed as an alternative and potentially immune-privileged site for pancreatic islet transplantation. The aim of the present study was to assess the survival and rejection mechanisms of free and encapsulated xenogeneic islets transplanted into the medullary cavity of the femur, or under the kidney capsule of streptozotocin-induced diabetic C57BL/6 mice. The median survival of free rat islets transplanted into the bone marrow or under the kidney capsule was 9 and 14 days, respectively, whereas that of free human islets was shorter, 7 days (bone marrow) and 10 days (kidney capsule). Infiltrating CD8+ T cells and redistributed CD4+ T cells, and macrophages were detected around the transplanted islets in bone sections. Recipient mouse splenocytes proliferated in response to donor rat stimulator cells. One month after transplantation under both kidney capsule or into bone marrow, encapsulated rat islets had induced a similar degree of fibrotic reaction and still contained insulin positive cells. In conclusion, we successfully established a small animal model for xenogeneic islet transplantation into the bone marrow. The rejection of xenogeneic islets was associated with local and systemic T cell responses and macrophage recruitment. Although there was no evidence for immune-privilege, the bone marrow may represent a feasible site for encapsulated xenogeneic islet transplantation.

Meier, Raphael P. H.; Seebach, Jorg D.; Morel, Philippe; Mahou, Redouan; Borot, Sophie; Giovannoni, Laurianne; Parnaud, Geraldine; Montanari, Elisa; Bosco, Domenico; Wandrey, Christine; Berney, Thierry; Buhler, Leo H.; Muller, Yannick D.

2014-01-01

80

Use of a calcium sulfate-calcium phosphate synthetic bone graft composite in the surgical management of primary bone tumors.  

PubMed

Benign primary bone tumors are commonly treated with intralesional curettage with or without the use of surgical adjuvants. The reconstructive approach to the resulting contained bone defects is controversial, and clinical practice is varied. Synthetic bone substitutes may provide early mechanical support while minimizing the risks of disease transmission, nonunion, infection, and donor-site morbidity. Limited data exists regarding the use of calcium sulfate-calcium phosphate composite bone substitute for this purpose. The authors retrospectively reviewed the clinical outcomes of 24 patients with benign primary bone tumors who underwent intralesional curettage followed by reconstruction with a calcium sulfate-calcium phosphate composite bone substitute. Mean follow-up was 23 months. The most common diagnosis was giant cell tumor of bone. Six patients had upper-extremity tumors and 18 had lower-extremity tumors. Mean preoperative radiographic tumor volume was 41.0 cm(3). Mean volume of PRO-DENSE (Wright Medical Technology, Arlington, Tennessee) used in each patient was 15.6 cm(3). Mean time to full weight bearing for all patients was 7.3 weeks. Two patients sustained local tumor recurrences. No postoperative fractures occurred, and no complications occurred related to the use of the calcium sulfate-calcium phosphate composite. One case of deep infection occurred secondary to wound breakdown. The use of a calcium sulfate-calcium phosphate composite was associated with rapid biological integration and an early return to activities of daily living, with no composite-related complications. This technique is a viable option in the reconstruction of cavitary bone defects following intralesional curettage of primary benign bone tumors. PMID:23380017

Evaniew, Nathan; Tan, Victoria; Parasu, Naveen; Jurriaans, Erik; Finlay, Karen; Deheshi, Benjamin; Ghert, Michelle

2013-02-01

81

Expression and methylation analysis of p15 and p16 in mouse bone marrow cells exposed to 1,4-benzoquinone.  

PubMed

Benzene is an important industrial chemical. It is also an environmental pollutant recognized as a human carcinogen. Both prenatal and adult exposures to benzene are associated with the development of leukemia. To understand the mechanism of benzene-induced epigenetic variations, we investigated the expression and methylation patterns of CpG (phosphodiester bond between cytosine and guanine) islands in p15 and p16 promoter regions in 1,4-benzoquinone (1,4-BQ)-treated primary cultivated C57BL/6J mouse bone marrow cells in vitro. The cell toxicity of 1,4-BQ was evaluated by cell viability test, real-time PCR was used to measure the mRNA expression levels, and bisulfite sequencing PCR (BSP) was used to look into the methylation patterns. The cell viability test indicates that 1,4-BQ exhibited a dose-dependent toxicity to mouse bone marrow cells. After a 24-h exposure to 1,4-BQ at final concentrations of 0, 0.1, 1, and 10 ?mol/L, the mRNA expression of p15 and p16 decreased with the increase in 1,4-BQ concentration. The BSP results gathered from the exposure and the control groups were the same. In summary, despite the observation that short-term exposure to 1,4-BQ primary cultivated mouse bone marrow cells decreased the p15 and p16 transcripts, with no influence by their gene promoter methylation. PMID:22027503

Tian, J-F; Peng, C-H; Yu, X-Y; Yang, X-J; Yan, Hong-Tao

2012-07-01

82

Transgenic mouse model for neurocristopathy: Schwannomas and facial bone tumors.  

PubMed Central

We have characterized a strain of double transgenic mice with simian virus 40 large tumor antigen and prokaryotic lacZ under the control of the myelin basic protein promoter that develops spindle-cell sarcomas and osteogenic sarcomas at 5-7 months of age. Although poorly differentiated, the spindle-cell sarcomas were characterized as malignant Schwannomas based on their neural association, the presence of basal lamina, and expression of Schwann cell-specific genes. The osteogenic sarcomas were often multiple and appeared predominantly in the facial bones, less frequently in the ribs and vertebral column, and only rarely in the appendicular skeleton. Benign osteoblastic lesions were often observed adjacent to these sarcomas. Both the osteoblastic cells in the facial skeleton and Schwann cells are regarded as neural crest derivatives. The biological properties and anatomical location of these tumors suggest that they may share a common origin from the neural crest or its derivatives. R.P. Bolande [Hum. Pathol. (1974) 5, 409-429] introduced the term neurocristopathy as a unifying concept to describe such lesions arising from the neural crest or its derivatives. Cell lines established from both bone and Schwann cell tumors arising in these transgenic mice express simian virus 40 large tumor antigen mRNA as well as functional large tumor antigen. Such cell lines are potentially valuable in the search for markers that identify mammalian neural crest derivatives. Images Fig. 1 Fig. 2 Fig. 3

Jensen, N A; Rodriguez, M L; Garvey, J S; Miller, C A; Hood, L

1993-01-01

83

Proinflammatory Cytokine Response and Viral Replication in Mouse Bone Marrow Derived Macrophages Infected with Influenza H1N1 and H5N1 Viruses  

PubMed Central

The pathogenesis of human influenza H5N1 virus infection remains poorly understood and controversial. Cytokine dysregulation in human infection has been hypothesized to contribute to disease severity. We developed in vitro cultures of mouse bone marrow derived macrophages (BMDM?) from C57BL/6N mouse to compare influenza A (H5N1 and H1N1) virus replication and pro-inflammatory cytokine and chemokine responses. While both H1N1 and H5N1 viruses infected the mouse bone marrow derived macrophages, only the H1N1 virus had showed evidence of productive viral replication from the infected cells. In comparison with human seasonal influenza H1N1 (A/HK/54/98) and mouse adapted influenza H1N1 (A/WSN/33) viruses, the highly pathogenic influenza H5N1 virus (A/HK/483/97) was a more potent inducer of the chemokine, CXCL 10 (IP-10), while there was not a clear differential TNF-? protein expression pattern. Although human influenza viruses rarely cause infection in mice without prior adaption, the use of in vitro cell cultures of primary mouse cells is of interest, especially given the availability of gene-defective (knock-out) mice for specific genes.

Chan, Renee W. Y.; Leung, Connie Y. H.; Nicholls, John M.; Peiris, J. S. Malik; Chan, Michael C. W.

2012-01-01

84

Identification of the Earliest B Lineage Stage in Mouse Bone Marrow  

Microsoft Academic Search

We have identified a very early stage of B lineage cells in the CD45R (B220)+CD24 (HSA)? pre-pro-B fraction of mouse bone marrow delineated by expression of AA4.1, a molecule found on stem cells and early B lineage cells. These cells are B lineage precursors based on their capacity to generate B lineage cells rapidly in stromal-dependent culture and their expression

Yue-Sheng Li; Robert Wasserman; Kyoko Hayakawa; Richard R. Hardy

1996-01-01

85

Primary bone marrow B-cell non-Hodgkin's lymphoma successfully treated with R-CHOP.  

PubMed

Primary isolated bone marrow disease as a presenting feature of lymphoma is very rare. We describe the case of a Chinese with isolated bone marrow small B-cell lymphoma as a first manifestation. A 55-year old woman was admitted to our hospital with fever. Her peripheral blood smear and laboratory findings were suggestive of bicytopenia. Bone marrow specimen showed diffusely distributed small-sized lymphocytes. Combined with immunophenotypic and chromosomal analysis, a diagnosis of primary bone marrow B-cell non-Hodgkin's lymphoma was made. The patient was treated with R-CHOP (rituximab and cyclophosphamide, epirubicin, vindesine, and prednisone) regimen for six cycles. She had complete remission and is still alive without relapse. We concluded that primary bone marrow mature small B-cell lymphoma is a rare but distinctive subtype of lymphoma. The prognosis for this entity is poor but rituximab-based treatment is promising for improving its outcomes. PMID:24171336

Qian, Liren; Zhang, Zhi; Shen, Jianliang; Liu, Yi

2013-01-01

86

OPG knockout mouse teeth display reduced alveolar bone mass and hypermineralization in enamel and dentin.  

PubMed

Recent studies showed that local injection or upregulation of OPG gene would result in early temporal retardation of tooth development. It was assumed that this retardation might cause defective tooth mineralization and pulp formation as the long-term effects. However, since those OPG treatments were transient, any possible long-term effects of OPG addition could not be assessed previously. In the present study, a high-resolution microCT was used to evaluate the long-term effect of OPG gene deprivation on the mineralization and morphology of mouse tooth. Our results showed that the mineralization of alveolar bone in OPG(-/-) mouse tooth was decreased while those of enamel and dentin were increased, compared with the wild-type (WT) group. The labial and lingual dentin thicknesses of OPG(-/-) group were significantly higher and with larger area in enamel and dentin than those of WT group. The size of pulp chamber was also substantially decreased in OPG(-/-) mouse incisor. Different responses in mineralization and morphogenesis to OPG gene deprivation were found between bone and tooth. These effects may be independent of the early odontogenesis, and further studies are warranted to investigate the molecular mechanism of the effect of OPG gene expression on bone formation and later tooth development. PMID:20233613

Sheng, Zhi-Feng; Ye, Wei; Wang, Jie; Li, Chun-Hai; Liu, Jiang-Hua; Liang, Qing-Chun; Li, Shan; Xu, Kang; Liao, Er-Yuan

2010-04-01

87

Magnetic assembly-mediated enhancement of differentiation of mouse bone marrow cells cultured on magnetic colloidal assemblies.  

PubMed

Here we reported an interesting phenomenon that the field-induced assemblies of magnetic nanoparticles can promote the differentiation of primary mouse bone marrow cells into osteoblasts. The reason was thought to lie in the remnant magnetic interaction inside the assemblies which resulted from the magnetic field-directed assembly. Influence of the assemblies on the cells was realized by means of interface effect rather than the internalization effect. We fabricated a stripe-like assemblies array on the glass plate and cultured cells on this surface. We characterized the morphology of assemblies and measured the mechanic property as well as the magnetic property. The cellular differentiation was measured by staining and quantitative PCR. Finally, Fe uptake was excluded as the reason to cause the phenomenon. PMID:24874764

Sun, Jianfei; Liu, Xuan; Huang, Jiqing; Song, Lina; Chen, Zihao; Liu, Haoyu; Li, Yan; Zhang, Yu; Gu, Ning

2014-01-01

88

Magnetic assembly-mediated enhancement of differentiation of mouse bone marrow cells cultured on magnetic colloidal assemblies  

PubMed Central

Here we reported an interesting phenomenon that the field-induced assemblies of magnetic nanoparticles can promote the differentiation of primary mouse bone marrow cells into osteoblasts. The reason was thought to lie in the remnant magnetic interaction inside the assemblies which resulted from the magnetic field-directed assembly. Influence of the assemblies on the cells was realized by means of interface effect rather than the internalization effect. We fabricated a stripe-like assemblies array on the glass plate and cultured cells on this surface. We characterized the morphology of assemblies and measured the mechanic property as well as the magnetic property. The cellular differentiation was measured by staining and quantitative PCR. Finally, Fe uptake was excluded as the reason to cause the phenomenon.

Sun, Jianfei; Liu, Xuan; Huang, Jiqing; Song, Lina; Chen, Zihao; Liu, Haoyu; Li, Yan; Zhang, Yu; Gu, Ning

2014-01-01

89

COLLAGEN MUTATION CAUSES CHANGES OF THE MICRODAMAGE MORPHOLOGY IN BONE OF AN OI MOUSE MODEL  

PubMed Central

Previous studies have postulated that ultrastructural changes may alter the pattern and capacity of microdamage accumulation in bone. Using an osteogenesis imperfecta (OI) mouse model, this study was performed to investigate the correlation of collagen mutation with the microdamage morphology and the associated brittleness of bone. In this study, femurs from mild OI and wild type mice were fatigued under four-point bending to create microdamage in the specimens. Then, the microdamage morphology of these specimens was examined using the bulk-staining technique with basic fuchsin. Similar with the results of previous studies, it was observed that linear microcracks were formed more easily in compression, whereas diffuse damage was induced more readily in tension for both wild-type and mild-type mice. However, less diffuse damage was found in the tensile side of mild OI mouse femurs (collagen mutation) compared with those of wild type mice, showing that the microdamage morphology is correlated to the brittleness of bone. The results of this study provide direct evidence that supports the prediction made by the previous numerical simulation studies, suggesting that microdamage morphology in bone is significantly correlated with the integrity of the collagen phase.

Dong, X. Neil; Zoghi, Mahyar; Ran, Qitao; Wang, Xiaodu

2010-01-01

90

Participation of Adult Mouse Bone Marrow Cells in Reconstitution of Skin  

PubMed Central

Results of recent studies have indicated that bone marrow cells can differentiate into various cells of ectodermal, mesodermal, and endodermal origins when transplanted into the body. However, the problems associated with those experiments such as the long latent period, rareness of the event, and difficulty in controlling the processes have hampered detailed mechanistic studies. In the present study, we examined the potency of mouse bone marrow cells to differentiate into cells comprising skin tissues using a skin reconstitution assay. Bone marrow cells from adult green fluorescent protein (GFP)-transgenic mice were transplanted in a mixture of embryonic mouse skin cells (17.5 days post-coitus) onto skin defects made on the backs of nude mice. Within 3 weeks, fully differentiated skin with hair was reconstituted. GFP-positive cells were found in the epidermis, hair follicles, sebaceous glands, and dermis. The localization and morphology of the cells, results of immunohistochemistry, and results of specific staining confirmed that the bone marrow cells had differentiated into epidermal keratinocytes, sebaceous gland cells, follicular epithelial cells, dendritic cells, and endothelial cells under the present conditions. These results indicate that this system is suitable for molecular and cellular mechanistic studies on differentiation of stem cells to various epidermal and dermal cells.

Kataoka, Ken; Medina, Reinhold J.; Kageyama, Tomofumi; Miyazaki, Masahiro; Yoshino, Tadashi; Makino, Teruhiko; Huh, Nam-ho

2003-01-01

91

Chromosome behaviour during early meiotic prophase of mouse primary spermatocytes  

Microsoft Academic Search

A correlative light- and electron microscopical study has yielded both a description of chromosome behaviour during early meiotic prophase in mouse spermatocytes, and an accurate timing of the zygotene and early pachytene stage. The light microscopic part of the study consists of an analysis of orcein stained or C-banded, air-dried preparations, and series of separately squashed sections of seminiferous tubules.

J. L. Oud; A. H. H. Reutlinger

1981-01-01

92

Malathion and fenvalerate induce micronuclei in mouse bone marrow cells.  

PubMed

Health effects of pesticides are a major public health concern. In this study, the genotoxic effects of two commonly-used pesticides, malathion, and fenvalerate, were investigated in mice in vivo. Induction of micronuclei in bone marrow cells was used as the test parameter following exposure to 2.5, 5 or 10 mg/kg malathion by intraperitoneal (i.p.) or per oral (p.o.) exposure. Exposure by both routes was found to cause a significant increase in micronucleated polychromatic erythrocytes (PCEs) in a dose-dependent manner (r = 0.9769; P < 0.05). The highest dose (10 mg/kg) induced significant (P < 0.05) cytotoxicity. In contrast, fenvalerate caused an increase in micronucleated PCEs only at higher doses (10 and 20 mg/kg) via i.p. injection, and was not associated with cytotoxicity. A significant dose-response correlation was not observed in the dose ranges tested for fenvalerate (r = 0.8704; P > 0.05). The results suggest that technical grade malathion is a genotoxic agent. In contrast, technical grade fenvalerate appears to be a potent genotoxic agent, but this observation should be confirmed with further investigation(s). PMID:21538555

Giri, A; Giri, S; Sharma, G D

2011-10-01

93

QUANTITATIVE STUDIES ON THE STIMULATION OF MOUSE BONE MARROW COLONY GROWTH IN VITRO BY NORMAL HUMAN URINE  

Microsoft Academic Search

Normal human urine was shown to stimulate colony formation in vitro by mouse bone marrow cells. Linear relationships were demonstrated between urine dose and the number and size of colonies stimulated to develop from a standard number of bone marrow cells. These criteria provide an assay system for quantifying levels of colony stimulating factor In human urine. A survey of

D Metcalf; ER Stanley

1969-01-01

94

Return to Primary Service Among Bone Marrow Transplant Rehabilitation Inpatients: An Index for Predicting Outcomes Authors  

PubMed Central

Objective To assess rehabilitation inpatient risk of return to primary service in bone marrow transplant patients. Design Retrospective review. Setting Inpatient rehabilitation unit within a tertiary referral based cancer center Participants All bone marrow transplant patients (131) who were admitted a total of 147 times to inpatient rehabilitation between January 1, 2002, and April 30, 2010. Interventions None. Main Outcome Measures We analyzed return to primary service and demographic information, cancer characteristics, medications, hospital admission characteristics, and laboratory values. Results 41% (61/147) of bone marrow transplant admissions were transferred from the inpatient rehabilitation unit back to the primary service. Of those transferred back, 38% (23/61) died after being transferred back to the primary service. Significant or near significant relationships were found for a platelet count < 43,000 per microliter (p<.01), a creatinine level > 0.9 milligrams/deciliter (p<.01), the presence of an antiviral agent (p=.0501), the presence of an antibacterial agent (p=.0519), the presence of an antifungal agent (p<.05) and leukemia, lymphoma or multiple myeloma diagnosis (p<.05). Using five of these factors the Return to Primary-Bone Marrow Transplant (RTP-BMT) index was formulated to determine the likelihood of return to the primary team. Conclusion Bone marrow transplant patients have a high rate of transfer from the inpatient rehabilitation unit back to the primary service. The RTP-BMT score can be a useful tool to help clinicians predict the likelihood of return to the primary acute care service.

Fu, Jack Brian; Lee, Jay; Smith, Dennis W.; Guo, Ying; Bruera, Eduardo

2012-01-01

95

?-Actinin-3 deficiency is associated with reduced bone mass in human and mouse.  

PubMed

Bone mineral density (BMD) is a complex trait that is the single best predictor of the risk of osteoporotic fractures. Candidate gene and genome-wide association studies have identified genetic variations in approximately 30 genetic loci associated with BMD variation in humans. ?-Actinin-3 (ACTN3) is highly expressed in fast skeletal muscle fibres. There is a common null-polymorphism R577X in human ACTN3 that results in complete deficiency of the ?-actinin-3 protein in approximately 20% of Eurasians. Absence of ?-actinin-3 does not cause any disease phenotypes in muscle because of compensation by ?-actinin-2. However, ?-actinin-3 deficiency has been shown to be detrimental to athletic sprint/power performance. In this report we reveal additional functions for ?-actinin-3 in bone. ?-Actinin-3 but not ?-actinin-2 is expressed in osteoblasts. The Actn3(-/-) mouse displays significantly reduced bone mass, with reduced cortical bone volume (-14%) and trabecular number (-61%) seen by microCT. Dynamic histomorphometry indicated this was due to a reduction in bone formation. In a cohort of postmenopausal Australian women, ACTN3 577XX genotype was associated with lower BMD in an additive genetic model, with the R577X genotype contributing 1.1% of the variance in BMD. Microarray analysis of cultured osteoprogenitors from Actn3(-/-) mice showed alterations in expression of several genes regulating bone mass and osteoblast/osteoclast activity, including Enpp1, Opg and Wnt7b. Our studies suggest that ACTN3 likely contributes to the regulation of bone mass through alterations in bone turnover. Given the high frequency of R577X in the general population, the potential role of ACTN3 R577X as a factor influencing variations in BMD in elderly humans warrants further study. PMID:21784188

Yang, Nan; Schindeler, Aaron; McDonald, Michelle M; Seto, Jane T; Houweling, Peter J; Lek, Monkol; Hogarth, Marshall; Morse, Alyson R; Raftery, Joanna M; Balasuriya, Dominic; MacArthur, Daniel G; Berman, Yemima; Quinlan, Kate G R; Eisman, John A; Nguyen, Tuan V; Center, Jacqueline R; Prince, Richard L; Wilson, Scott G; Zhu, Kathy; Little, David G; North, Kathryn N

2011-10-01

96

Targeting of Primary Breast Cancers and Metastases in a Transgenic Mouse Model Using Rationally Designed Multifunctional SPIONs  

PubMed Central

Breast cancer remains one of the most prevalent and lethal malignancies in women. The inability to diagnose small volume metastases early has limited effective treatment of stage 4 breast cancer. Here we report the rational development and use of a multifunctional superparamagnetic iron oxide nanoparticle (SPION) for targeting metastatic breast cancer in a transgenic mouse model and imaging with magnetic resonance (MR). SPIONs coated with a copolymer of chitosan and polyethylene glycol (PEG) were labeled with a fluorescent dye for optical detection and conjugated with a monoclonal antibody against the neu receptor (NP-neu). SPIONs labeled with mouse IgG were used as a non-targeting control (NP-IgG). These SPIONs had desirable physiochemical properties for in vivo applications such as near neutral zeta potential and hydrodynamic size around 40 nm, and were highly stable in serum containing medium. Only NP-neu showed high uptake in neu expressing mouse mammary carcinoma (MMC) cells which was reversed by competing free neu antibody, indicating their specificity to the neu antigen. In vivo, NP-neu was able to tag primary breast tumors and significantly, only NP-neu bound to spontaneous liver, lung, and bone marrow metastases in a transgenic mouse model of metastatic breast cancer, highlighting the necessity of targeting for delivery to metastatic disease. The SPIONs provided significant contrast enhancement in MR images of primary breast tumors; thus, they have the potential for MRI detection of micrometastases, and provide an excellent platform for further development of an efficient metastatic breast cancer therapy.

Kievit, Forrest M.; Stephen, Zachary R.; Veiseh, Omid; Arami, Hamed; Wang, Tingzhong; Lai, Vy P.; Park, James O.; Ellenbogen, Richard G.; Disis, Mary L.; Zhang, Miqin

2012-01-01

97

Ex vivo expansions and transplantations of mouse bone marrow-derived hematopoietic stem/progenitor cells.  

PubMed

To examine the effects of co-culture with bone marrow mesenchymal stem cells on expansion of hematopoietic stem/progenitor cells and the capacities of rapid neutrophil engraftment and hematopoietic reconstitution of the expanded cells, we expanded mononuclear cells (MNCs) and CD34+/c-kit+ cells from mouse bone marrow and transplanted the expanded cells into the irradiated mice. MNCs were isolated from mouse bone marrow and CD34+/c-kit+ cells were selected from MNCs by using MoFlo Cell Sorter. MNCs and CD34+/c-kit+ cells were co-cultured with mouse bone marrow-derived mesenchymal stem cells (MSCs) under a two-step expansion. The expanded cells were then transplanted into sublethally irradiated BDF1 mice. Results showed that the co-culture with MSCs resulted in expansions of median total nucleated cells, CD34+ cells, GM-CFC and HPP-CFC respectively by 10.8-, 4.8-, 65.9- and 38.8-fold for the mononuclear cell culture, and respectively by 76.1-, 2.9-, 71.7- and 51.8-fold for the CD34+/c-kit+ cell culture. The expanded cells could rapidly engraft in the sublethally irradiated mice and reconstitute their hematopoiesis. Co-cultures with MSCs in conjunction with two-step expansion increased expansions of total nucleated cells, GM-CFC and HPP-CFC, which led us to conclude MSCs may create favorable environment for expansions of hematopoietic stem/progenitor cells. The availability of increased numbers of expanded cells by the co-culture with MSCs may result in more rapid engraftment of neutrophils following infusion to transplant recipients. PMID:14674026

Wang, Jin-fu; Wu, Yi-fan; Harrintong, Jenny; McNiece, Ian K

2004-02-01

98

Newly developed multiple myeloma in a patient with primary T-cell lymphoma of bone.  

PubMed

Primary non-Hodgkin's lymphoma of bone (PLB) is rare, and generally presents as a single extensive and destructive bone lesion. Histopathologically, most cases present as diffuse large B-cell lymphoma, and T-cell lymphoma is rare. By contrast, multiple myeloma is a disease defined as the neoplastic proliferation of a single clone of plasma cells producing a monoclonal immunoglobulin. We report a case of multiple myeloma that developed during treatment of PLB in a type of T-cell. A 48-yr-old man was diagnosed as T-cell PLB, stage IE, 18 months ago. The patient received the chemoradiotherapy and salvage chemotherapy for PLB. However, the lymphoma progressed with generalized bone pain, and laboratory findings showed bicytopenia and acute renal failure. On bone marrow biopsy, the patient was diagnosed as having multiple myeloma newly developed with primary T-cell lymphoma of bone. In spite of chemotherapy, the patient died of renal failure. PMID:18583898

Hwang, Jun-Eul; Cho, Sang-Hee; Kim, Ok-Ki; Shim, Hyun-Jeong; Lee, Se-Ryeon; Ahn, Jae-Sook; Yang, Duk-Hwan; Kim, Yeo-Kyeoung; Lee, Je-Jung; Kim, Hyeoung-Joon; Chung, Ik-Joo

2008-06-01

99

Newly Developed Multiple Myeloma in a Patient with Primary T-Cell Lymphoma of Bone  

PubMed Central

Primary non-Hodgkin's lymphoma of bone (PLB) is rare, and generally presents as a single extensive and destructive bone lesion. Histopathologically, most cases present as diffuse large B-cell lymphoma, and T-cell lymphoma is rare. By contrast, multiple myeloma is a disease defined as the neoplastic proliferation of a single clone of plasma cells producing a monoclonal immunoglobulin. We report a case of multiple myeloma that developed during treatment of PLB in a type of T-cell. A 48-yr-old man was diagnosed as T-cell PLB, stage IE, 18 months ago. The patient received the chemoradiotherapy and salvage chemotherapy for PLB. However, the lymphoma progressed with generalized bone pain, and laboratory findings showed bicytopenia and acute renal failure. On bone marrow biopsy, the patient was diagnosed as having multiple myeloma newly developed with primary T-cell lymphoma of bone. In spite of chemotherapy, the patient died of renal failure.

Hwang, Jun-Eul; Cho, Sang-Hee; Kim, Ok-Ki; Shim, Hyun-Jeong; Lee, Se-Ryeon; Ahn, Jae-Sook; Yang, Duk-Hwan; Kim, Yeo-Kyeoung; Lee, Je-Jung; Kim, Hyeoung-Joon

2008-01-01

100

Age-Related Changes in Bone Mass in the Senescence-Accelerated Mouse (SAM)  

PubMed Central

Age-related changes of the femoral bone mass in several strains of the senescence-accelerated mouse (SAM) were investigated. Microdensitometrically, all strains exhibited essentially the same patterns of age changes, that is, bone mass corrected by the diameter of the shaft reached the peak value when the mice were 4 or 5 months of age and then fell linearly with age up to over 20 months of age. Two strains, SAM-R/3 and SAM-P/6, which originated from the same ancestry on pedigree, had a significantly lower peak bone mass than other strains (SAM-R/1, SAM-R/2, SAM-P/1, and SAM-P/2). On the other hand, the strains with a low peak bone mass had the same rate of decrease as other strains. Mineral and collagen contents per dry weight of bone showed little difference among the strains. Histologic studies of tibia, femur, and lumbar spine revealed that the osteopenia was not due to osteomalacia but, rather, to osteoporosis. The elderly mice in these two strains were prone to fracture, thus should be important models for study of senile osteoporosis seen clinically. ImagesFigure 7Figure 8

Matsushita, Mutsumi; Tsuboyama, Tadao; Kasai, Ryuichi; Okumura, Hideo; Yamamuro, Takao; Higuchi, Keiichi; Higuchi, Kayoko; Kohno, Atsuko; Yonezu, Tomonori; Utani, Atsushi; Umezawa, Makiko; Takeda, Toshio

1986-01-01

101

Assessment of lamellar level properties in mouse bone utilizing a novel spherical nanoindentation data analysis method.  

PubMed

In this work, we demonstrate the viability of using our recently developed data analysis procedures for spherical nanoindentation in conjunction with Raman spectroscopy for studying lamellar-level correlations between the local composition and local mechanical properties in mouse bone. Our methodologies allow us to convert the raw load-displacement datasets to much more meaningful indentation stress-strain curves that accurately capture the loading and unloading elastic moduli, the indentation yield points, as well as the post-yield characteristics in the tested samples. Using samples of two different inbred mouse strains, A/J and C57BL/6J (B6), we successfully demonstrate the correlations between the mechanical information obtained from spherical nanoindentation measurements to the local composition measured using Raman spectroscopy. In particular, we observe that a higher mineral-to-matrix ratio correlated well with a higher local modulus and yield strength in all samples. Thus, new bone regions exhibited lower moduli and yield strengths compared to more mature bone. The B6 mice were also found to exhibit lower modulus and yield strength values compared to the more mineralized A/J strain. PMID:22842281

Pathak, Siddhartha; Vachhani, Shraddha J; Jepsen, Karl J; Goldman, Haviva M; Kalidindi, Surya R

2012-09-01

102

Abnormal bone formation induced by implantation of osteosarcoma-derived bone-inducing substance in the X-linked hypophosphatemic mouse  

Microsoft Academic Search

The X-linked hypophosphatemic mouse (Hyp) has been proposed as a model for the human familial hypophosphatemia (the most common form of vitamin D-resistant rickets). An osteosarcoma-derived bone-inducing substance was subcutaneously implanted into the Hyp mouse. The implant was consistently replaced by cartilage tissue at 2 weeks after implantation. The cartilage matrix seemed to be normal, according to the histological examination,

H. Yoshikawa; K. Masuhara; K. Takaoka; K. Ono; H. Tanaka; Y. Seino

1985-01-01

103

Basic fibroblast growth factor supports expansion of mouse compact bone-derived mesenchymal stem cells (MSCs) and regeneration of bone from MSC in vivo.  

PubMed

Some progress has been made in development of methods to regenerate bone from cultured cells, however no method is put to practical use. Here, we developed methods to isolate, purify, and expand mesenchymal stem cells (MSCs) from mouse compact bone that may be used to regenerate bone in vivo. These cells were maintained in long-term culture and were capable of differentiating along multiple lineages, including chondrocyte, osteocyte, and adipocyte trajectories. We used standard cell isolation and culture methods to establish cell cultures from mouse compact bone and bone marrow. Cultures were grown in four distinct media to determine the optimal composition of culture medium for bone-derived MSCs. Putative MSCs were subjected to flow cytometry, alkaline phosphatase assays, immunohistochemical staining, and several differentiation assays to assess cell identity, protein expression, and developmental potential. Finally, we used an in vivo bone formation assay to determine whether putative MSCs were capable of regenerating bone. We found that compact bone of mice was a better source of MCSs than the bone marrow, that growth in plastic flasks served to purify MSCs from hematopoietic cells, and that MSCs grown in basic fibroblast growth factor (bFGF)-conditioned medium were, based on multiple criteria, superior to those grown in leukemia inhibitory factor-conditioned medium. Moreover, we found that the MSCs isolated from compact bone and grown in bFGF-conditioned medium were capable of supporting bone formation in vivo. The methods and results described here have implications for understanding MSC biology and for clinical purpose. PMID:22203245

Yamachika, Eiki; Tsujigiwa, Hidetsugu; Matsubara, Masakazu; Hirata, Yasuhisa; Kita, Kenichiro; Takabatake, Kiyofumi; Mizukawa, Nobuyoshi; Kaneda, Yoshihiro; Nagatsuka, Hitoshi; Iida, Seiji

2012-04-01

104

Changes in bone micro-architecture and biomechanical properties in the th3 thalassemia mouse are associated with decreased bone turnover and occur during the period of bone accrual  

PubMed Central

Osteoporosis and fractures occur frequently in patients with beta thalassemias, a group of congenital hemolytic anemias characterized by decreased synthesis of the beta chain of hemoglobin. In this study, we determined the bone abnormalities of the th3 thalassemia mouse, generated by deletion of the mouse beta chain genes. The heterozygote th3/+ mouse has moderate anemia, and serves as a model of beta thalassemia intermedia (TI), which represents the mild thalassemia phenotype. The th3/th3 mouse has lethal anemia and is a model of beta thalassemia major (TM), which is characterized by life-threatening anemia requiring regular transfusions to sustain life. Compared to controls: i) Micro-CT of trabecular bone showed decreased bone volume fraction, number of trabeculae and trabecular thickness in both th3/+ and th3/th3 (p<0.05). ii) Cortical bone analysis showed thinner cortices and increased marrow area in th3/+ animals (p<0.05). iii) Micro-CT abnormalities in th3/+ mice were present by 2 months and did not worsen with age. iv) Histomorphometry was significant for decreased bone formation and resorption in both th3/+ and th3/th3. Similarly, cathepsin K and osteocalcin expression from bone of both th3/+and th3/th3 animals was reduced (p<0.05). vi) Biomechanics showed reduced maximum load, maximum moment and structural stiffness in both th3/+and th3/th3 (p<0.01). In conclusion, the th3 mouse model of thalassemia manifests bone changes reminiscent of those in humans, and can be used for further bone studies in thalassemia. Bone changes are associated with decreased bone turnover, and develop early on during the period of bone accrual.

Vogiatzi, Maria G.; Tsay, Jaime; Verdelis, Kostas; Rivella, Stefano; Grady, Robert W; Doty, Stephen; Giardina, Patricia J; Boskey, Adele L

2010-01-01

105

Cardiopulmonary dysfunction in the Osteogenesis imperfecta mouse model Aga2 and human patients are caused by bone-independent mechanisms.  

PubMed

Osteogenesis imperfecta (OI) is an inherited connective tissue disorder with skeletal dysplasia of varying severity, predominantly caused by mutations in the collagen I genes (COL1A1/COL1A2). Extraskeletal findings such as cardiac and pulmonary complications are generally considered to be significant secondary features. Aga2, a murine model for human OI, was systemically analyzed in the German Mouse Clinic by means of in vivo and in vitro examinations of the cardiopulmonary system, to identify novel mechanisms accounting for perinatal lethality. Pulmonary and, especially, cardiac fibroblast of perinatal lethal Aga2/+ animals display a strong down-regulation of Col1a1 transcripts in vivo and in vitro, resulting in a loss of extracellular matrix integrity. In addition, dysregulated gene expression of Nppa, different types of collagen and Agt in heart and lung tissue support a bone-independent vicious cycle of heart dysfunction, including hypertrophy, loss of myocardial matrix integrity, pulmonary hypertension, pneumonia and hypoxia leading to death in Aga2. These murine findings are corroborated by a pediatric OI cohort study, displaying significant progressive decline in pulmonary function and restrictive pulmonary disease independent of scoliosis. Most participants show mild cardiac valvular regurgitation, independent of pulmonary and skeletal findings. Data obtained from human OI patients and the mouse model Aga2 provide novel evidence for primary effects of type I collagen mutations on the heart and lung. The findings will have potential benefits of anticipatory clinical exams and early intervention in OI patients. PMID:22589248

Thiele, Frank; Cohrs, Christian M; Flor, Armando; Lisse, Thomas S; Przemeck, Gerhard K H; Horsch, Marion; Schrewe, Anja; Gailus-Durner, Valerie; Ivandic, Boris; Katus, Hugo A; Wurst, Wolfgang; Reisenberg, Catherine; Chaney, Hollis; Fuchs, Helmut; Hans, Wolfgang; Beckers, Johannes; Marini, Joan C; Hrabé de Angelis, Martin

2012-08-15

106

Cardiopulmonary dysfunction in the Osteogenesis imperfecta mouse model Aga2 and human patients are caused by bone-independent mechanisms  

PubMed Central

Osteogenesis imperfecta (OI) is an inherited connective tissue disorder with skeletal dysplasia of varying severity, predominantly caused by mutations in the collagen I genes (COL1A1/COL1A2). Extraskeletal findings such as cardiac and pulmonary complications are generally considered to be significant secondary features. Aga2, a murine model for human OI, was systemically analyzed in the German Mouse Clinic by means of in vivo and in vitro examinations of the cardiopulmonary system, to identify novel mechanisms accounting for perinatal lethality. Pulmonary and, especially, cardiac fibroblast of perinatal lethal Aga2/+ animals display a strong down-regulation of Col1a1 transcripts in vivo and in vitro, resulting in a loss of extracellular matrix integrity. In addition, dysregulated gene expression of Nppa, different types of collagen and Agt in heart and lung tissue support a bone-independent vicious cycle of heart dysfunction, including hypertrophy, loss of myocardial matrix integrity, pulmonary hypertension, pneumonia and hypoxia leading to death in Aga2. These murine findings are corroborated by a pediatric OI cohort study, displaying significant progressive decline in pulmonary function and restrictive pulmonary disease independent of scoliosis. Most participants show mild cardiac valvular regurgitation, independent of pulmonary and skeletal findings. Data obtained from human OI patients and the mouse model Aga2 provide novel evidence for primary effects of type I collagen mutations on the heart and lung. The findings will have potential benefits of anticipatory clinical exams and early intervention in OI patients.

Thiele, Frank; Cohrs, Christian M.; Flor, Armando; Lisse, Thomas S.; Przemeck, Gerhard K. H.; Horsch, Marion; Schrewe, Anja; Gailus-Durner, Valerie; Ivandic, Boris; Katus, Hugo A.; Wurst, Wolfgang; Reisenberg, Catherine; Chaney, Hollis; Fuchs, Helmut; Hans, Wolfgang; Beckers, Johannes; Marini, Joan C.; Hrabe de Angelis, Martin

2012-01-01

107

ATRX dysfunction induces replication defects in primary mouse cells.  

PubMed

The chromatin remodeling protein ATRX, which targets tandem repetitive DNA, has been shown to be required for expression of the alpha globin genes, for proliferation of a variety of cellular progenitors, for chromosome congression and for the maintenance of telomeres. Mutations in ATRX have recently been identified in tumours which maintain their telomeres by a telomerase independent pathway involving homologous recombination thought to be triggered by DNA damage. It is as yet unknown whether there is a central underlying mechanism associated with ATRX dysfunction which can explain the numerous cellular phenomena observed. There is, however, growing evidence for its role in the replication of various repetitive DNA templates which are thought to have a propensity to form secondary structures. Using a mouse knockout model we demonstrate that ATRX plays a direct role in facilitating DNA replication. Ablation of ATRX alone, although leading to a DNA damage response at telomeres, is not sufficient to trigger the alternative lengthening of telomere pathway in mouse embryonic stem cells. PMID:24651726

Clynes, David; Jelinska, Clare; Xella, Barbara; Ayyub, Helena; Taylor, Stephen; Mitson, Matthew; Bachrati, Csanád Z; Higgs, Douglas R; Gibbons, Richard J

2014-01-01

108

Bone marrow metastasis in primary bronchial mucoepidermoid carcinoma: a case report  

PubMed Central

Primary bronchial mucoepidermoid carcinoma in the lung is relatively rare. It rarely presents with the highly malignant biological characteristic of bone marrow metastasis. We describe a case of this disease with bone marrow metastasis. A 56-year-old man with the primary manifestation of bone pain and bloodstained sputum had two abnormal shadows on the left inferior lobar bronchus and peripheral tissue of the lower lobe of the left lung, respectively. Computed tomography-guided percutaneous puncture biopsy and bone imaging confirmed the diagnosis of high-grade bronchial mucoepidermoid carcinoma with bone metastasis. However, the patient soon presented with progressive hemoglobin and platelet decline and severe multi-organ hemorrhage. Subsequently, we performed bone marrow aspiration and biopsy, which revealed malignant cells and necrosis. The patient deteriorated rapidly from the disease, and died on the 16th day of admission. We hope that this case report will increase awareness of the possibility of primary high-grade bronchial mucoepidermoid carcinoma metastasizing to the bone marrow, which might be a poor prognostic factor.

2014-01-01

109

Autograft reconstructions for bone defects in primary total knee replacement in severe varus knees  

PubMed Central

Background: Large posteromedial defects encountered in severe varus knees during primary total knee arthroplasty can be treated by cementoplasty, structural bone grafts or metallic wedges. The option is selected depending upon the size of the defect. We studied the outcome of autograft (structural and impaction bone grafting) reconstruction of medial tibial bone defects encountered during primary total knee replacement in severe varus knees. Materials and Methods: Out of 675 primary varus knees operated, bone defects in proximal tibia were encountered in 54 knees. Posteromedial defects involving 25-40% of the tibial condyle cut surface and measuring more than 5 mm in depth were grafted using a structural graft obtained from cut distal femur or proximal tibia in 48 knees. For larger, peripheral uncontained vertical defects in six cases, measuring >25 mm in depth and involving >40% cut surface of proximal tibial condyle, impaction bone grafting with a mesh support was used. Results: Bone grafts incorporated in 54 knees in 6 months. There was no graft collapse or stress fractures, loosening or nonunion. The average followup period was 7.8 years (range 5-10 years). We observed an average postoperative increase in the Knee Society Score from 40 to 90 points. There was improvement in the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores in terms of pain, stiffness and physical function during activities of daily living. Conclusion: Bone grafting for defects in primary total knee is justified as it is biological, available then and is cost effective besides preserving bone stock for future revisions. Structural grafts should be used in defects >5 mm deep and involving 25-40% of the cut proximal tibial condyle surface. For larger peripheral vertical defects, impaction bone grafting contained in a mesh should be done.

Kharbanda, Yatinder; Sharma, Mrinal

2014-01-01

110

Activity of iPMS and nPMS in mouse bone marrow micronucleus assays: comparison with mouse dominant lethal assay data.  

PubMed

isoPropyl methanesulphonate (iPMS) and its n-propyl analogue (nPMS) are shown to be active in mouse bone marrow micronucleus assays using male CBA, male and female (C3H/El X 102/E1)Fl and male and female Muta Mouse mice. iPMS was significantly more active than nPMS. No significant strain or gender differences were observed. These findings reflect the differences reported earlier for these two chemicals in mouse dominant lethal mutation assays. The earlier described dominant lethal assay data are represented schematically and discussed. PMID:8600355

Adler, I D; Tinwell, H; Kliesch, U; Ashby, J

1996-02-01

111

Mechanical Unloading of Mouse Bone in Microgravity Significantly Alters Cell Cycle Gene Set Expression  

NASA Astrophysics Data System (ADS)

Spaceflight factors, including microgravity and space radiation, have many detrimental short-term effects on human physiology, including muscle and bone degradation, and immune system dysfunction. The long-term progression of these physiological effects is still poorly understood, and a serious concern for long duration spaceflight missions. We hypothesized that some of the degenerative effects of spaceflight may be caused in part by an inability of stem cells to proliferate and differentiate normally resulting in an impairment of tissue regenerative processes. Furthermore, we hypothesized that long-term bone tissue degeneration in space may be mediated by activation of the p53 signaling network resulting in cell cycle arrest and/or apoptosis in osteoprogenitors. In our analyses we found that spaceflight caused significant bone loss in the weight-bearing bones of mice with a 6.3% reduction in bone volume and 11.9% decrease in bone thickness associated with increased osteoclastic activity. Along with this rapid bone loss we also observed alterations in the cell cycle characterized by an increase in the Cdkn1a/p21 cell cycle arrest molecule independent of Trp53. Overexpression of Cdkn1a/p21 was localized to osteoblasts lining the periosteal surface of the femur and chondrocytes in the head of the femur, suggesting an inhibition of proliferation in two key regenerative cell types of the femur in response to spaceflight. Additionally we found overexpression of several matrix degradation molecules including MMP-1a, 3 and 10, of which MMP-10 was localized to osteocytes within the shaft of the femur. This, in conjunction with 40 nm resolution synchrotron nano-Computed Tomography (nano-CT) observations of an increase in osteocyte lacunae cross-sectional area, perimeter and a decrease in circularity indicates a potential role for osteocytic osteolysis in the observed bone degeneration in spaceflight. To further investigate the genetic response of bone to mechanical unloading in spaceflight, we conducted genome wide microarray analysis of total RNA isolated from the mouse pelvis. Specifically, 16 week old mice were subjected to 15 days spaceflight onboard NASA's STS-131 space shuttle mission. The pelvis of the mice was dissected, the bone marrow was flushed and the bones were briefly stored in RNAlater. The pelvii were then homogenized, and RNA was isolated using TRIzol. RNA concentration and quality was measured using a Nanodrop spectrometer, and 0.8% agarose gel electrophoresis. Samples of cDNA were analyzed using an Affymetrix GeneChip\\S Gene 1.0 ST (Sense Target) Array System for Mouse and GenePattern Software. We normalized the ST gene arrays using Robust Multichip Average (RMA) normalization, which summarizes perfectly matched spots on the array through the median polish algorithm, rather than normalizing according to mismatched spots. We also used Limma for statistical analysis, using the BioConductor Limma Library by Gordon Smyth, and differential expression analysis to identify genes with significant changes in expression between the two experimental conditions. Finally we used GSEApreRanked for Gene Set Enrichment Analysis (GSEA), with Kolmogorov-Smirnov style statistics to identify groups of genes that are regulated together using the t-statistics derived from Limma. Preliminary results show that 6,603 genes expressed in pelvic bone had statistically significant alterations in spaceflight compared to ground controls. These prominently included cell cycle arrest molecules p21, and p18, cell survival molecule Crbp1, and cell cycle molecules cyclin D1, and Cdk1. Additionally, GSEA results indicated alterations in molecular targets of cyclin D1 and Cdk4, senescence pathways resulting from abnormal laminin maturation, cell-cell contacts via E-cadherin, and several pathways relating to protein translation and metabolism. In total 111 gene sets out of 2,488, about 4%, showed statistically significant set alterations. These alterations indicate significant impairment of normal cellular function in the mechanically unloaded envi

Blaber, Elizabeth; Dvorochkin, Natalya; Almeida, Eduardo; Kaplan, Warren; Burns, Brnedan

2012-07-01

112

Improved Culture-Based Isolation of Differentiating Endothelial Progenitor Cells from Mouse Bone Marrow Mononuclear Cells  

PubMed Central

Numerous endothelial progenitor cell (EPC)-related investigations have been performed in mouse experiments. However, defined characteristics of mouse cultured EPC have not been examined. We focused on fast versus slow adherent cell population in bone marrow mononuclear cells (BMMNCs) in culture and examined their characteristics. After 24 h-culture of BMMNCs, attached (AT) cells and floating (FL) cells were further cultured in endothelial differentiation medium separately. Immunological and molecular analyses exhibited more endothelial-like and less monocyte/macrophage-like characteristics in FL cells compared with AT cells. FL cells formed thick/stable tube and hypoxia or shear stress overload further enhanced these endothelial-like features with increased angiogenic cytokine/growth factor mRNA expressions. Finally, FL cells exhibited therapeutic potential in a mouse myocardial infarction model showing the specific local recruitment to ischemic border zone and tissue preservation. These findings suggest that slow adherent (FL) but not fast attached (AT) BMMNCs in culture are EPC-rich population in mouse.

Sekiguchi, Haruki; Ii, Masaaki; Jujo, Kentaro; Yokoyama, Ayumi; Hagiwara, Nobuhisa; Asahara, Takayuki

2011-01-01

113

EZH2 suppresses hepatocellular differentiation of mouse bone marrow mesenchymal stem cells.  

PubMed

Our previous studies have indicated that mouse bone marrow mesenchymal stem cells (mBMMSCs) have potential to differentiate into hepatocytes with high efficiency. Our study aimed to evaluate the role of the mouse histone methyltransferase enhancer of zeste homolog 2 gene (EZH2) in the hepatocellular differentiation of mBMMSCs. The mBMMSCs isolated from femurs and tibias were cultured in Iscove's modified Eagle's medium (IMEM) supplemented with 10% fetal bovine serum. Hepatocellular differentiation was induced by 20 ng/mL hepatocyte growth factor and 10 ng/mL fibroblast growth factor 4. The mouse histone methyltransferase EZH2 gene was introduced via PLenti-eGFP-EZH2 or PLenti-eGFP-NEO as a control. Hepatocellular-induced mBMMSCs showed lower expression of EZH2 and lower level of histone H3 lysine 27 trimethylation (H3K27me3) in the AFP and FOXa2 gene promoter regions compared to uninduced mBMMSCs. Introduction of EZH2 inhibited hepatocellular induction, reduced both the mRNA and protein levels of AFP and FOXa2, and increased the level of histone H3K27me3 in the AFP and FOXa2 gene promoter regions. In summary, the mouse histone methyltransferase EZH2 gene could suppress hepatocellular differentiation of mBMMSCs by increasing the level of H3K27me3 in the AFP and FOXa2 gene promoter regions. PMID:24737471

Lu, T; Sun, H; Lv, J; Yang, M F; Zhang, F; Qian, Y; Dong, X J

2014-01-01

114

Guidelines for histopathological specimen examination and diagnostic reporting of primary bone tumours  

PubMed Central

This review is intended to provide histopathologists with guidelines for clinical assessment, specimen handling and diagnostic reporting of benign and malignant primary bone tumours. Information from radiology, surgical, oncology and other clinical colleagues involved in the diagnosis and treatment of primary bone tumours should be properly assessed before undertaking a structured approach to specimen handling and histological reporting. This ensures that the information needed for planning appropriate treatment of these complex tumours is provided. Consistency in diagnostic evaluation with respect to both terminology and report content facilitates liaison at multidisciplinary bone tumour meetings and collaboration between cancer units and networks, as well as providing a common database for audit of the clinical, radiological and pathological aspects of bone tumours.

2011-01-01

115

Tenofovir treatment of primary osteoblasts alters gene expression profiles: implications for bone mineral density loss  

PubMed Central

There is strong clinical evidence that implicates tenofovir in the loss of bone mineral density during treatment of human immunodeficiency virus infection. In this study, we sought to test the hypothesis that tenofovir treatment of osteoblasts causes changes in the gene expression profile that would impact osteoblast function during bone formation. Primary osteoblasts were isolated and then treated with the tenofovir prodrug, tenofovir disoproxil fumarate (TDF). Total RNA from TDF-treated and untreated osteoblasts were extracted and used for microarray analysis to assess TDF-associated changes in the gene expression profile. Strikingly, the changes in gene expression profiles involved in cell signaling, cell cycle and amino acid metabolism, which would likely impact osteoblast function in bone formation. Our findings demonstrate for the first time that tenofovir treatment of primary osteoblasts results in gene expression changes that implicate loss of osteoblast function in tenofovir-associated bone mineral density loss.

Grigsby, Iwen F.; Pham, Lan; Mansky, Louis M.; Gopalakrishnan, Raj; Carlson, Ann E.; Mansky, Kim C.

2010-01-01

116

Bone morphogenetic proteins 4 and 2/7 induce osteogenic differentiation of mouse skin derived fibroblast and dermal papilla cells.  

PubMed

Heterotopic ossification is a pathological condition in which bone forms outside the skeletal system. It can also occur in skin, which is the case in some genetic disorders. In addition to precursor cells and the appropriate tissue environment, heterotopic ossification requires inductive signals such as bone morphogenetic proteins (BMP). BMPs are growth and differentiation factors that have the ability to induce cartilage and bone formation in ectopic sites. The objective of this study is to explore the effect of the BMP-4 homodimer and BMP-2/7 heterodimer on the osteogenic differentiation of primary mouse skin fibroblasts and hair follicle dermal papilla (DP) cells. Osteogenic differentiation was induced by osteogenic induction medium (OS) containing 10 nM dexamethasone. The effect of BMP-4 and BMP-2/7 was studied using alkaline phosphatase (ALP) and calcium assays after 1.5, 3 and 5 weeks of differentiation. Fibroblasts and DP cells were able to differentiate into osteoblast-like matrix mineralizing cells. The first visible sign of differentiation was the change of morphology from rounded to more spindle-shaped cells. BMP-4 and BMP-2/7 exposure elevated ALP activity and calcium production significantly more than OS alone. The osteogenic response to BMP-4 and BMP-2/7 was similar in fibroblasts, whereas, in DP cells, BMP-2/7 was more potent than BMP-4. OS alone could not induce osteogenic differentiation in DP cells. Clear and consistent results show that dermal fibroblasts and stem cells from the dermal papilla were capable of osteogenic differentiation. The BMP-2/7 heterodimer was significantly more effective on hair follicular dermal stem cell differentiation. PMID:24253465

Myllylä, Riina M; Haapasaari, Kirsi-Maria; Lehenkari, Petri; Tuukkanen, Juha

2014-02-01

117

Antibiotic bone cements: their use in routine primary total joint arthroplasty is justified.  

PubMed

The use of antibiotic bone cement in total hip arthroplasty (THA) is the standard of care in countries with long-standing national registries, as data from the registries demonstrates an improvement in survivorship by reducing the incidence of both septic and aseptic failures. There is reluctance in North America to embrace antibiotic bone cement, although the percentage of use is increasing. Reasons cited for not using antibiotic cement include lack of efficacy, adverse effects on mechanical properties, increased costs, bacterial resistance, and systemic toxicity. Little to no compelling data in the literature support these claims, and significant evidence refutes them, specifically: registry data and randomized controlled trials demonstrate a clear reduction in deep joint infections with the use of antibiotic bone cement; antibiotic bone cement has lower incidence of both septic and aseptic loosening, indicating no negative effect on mechanical properties; antibiotic bone cement is cost-effective, given its proven ability to reduce revision rates and prevent poor patient outcomes; there is no evidence to support bacterial resistance, and antibiotic bone cement may reduce the incidence of resistance; and there are no reported cases of systemic toxicity from manufacturer-prepared antibiotic cement in primary THA or total knee arthroplasty. Based on the strong evidence supporting the benefits of antibiotic bone cement and the lack of evidence against its use, it is difficult to justify why antibiotic cement is not the standard of care for primary cemented THA in North America. PMID:19751021

Dunbar, Michael J

2009-09-01

118

Identification of differentially expressed genes and their subpathways in recurrent versus primary bone giant cell tumors.  

PubMed

Giant cell tumor (GCT) of the bone is a benign but locally aggressive bone neoplasm with a strong tendency to develop local recurrent and metastatic disease. Thus, it provides a useful model system for the identification of biological mechanisms involved in bone tumor progression and metastasis. This study profiled 24 cases of recurrent versus primary bone GCT tissues using QuantiGene 2.0 Multiplex Arrays that included Human p53 80-Plex Panels and Human Stem Cell 80-Plex Panels. A total of 32 differentially expressed genes were identified, including the 20 most upregulated genes and the 12 most downregulated genes in recurrent GCT. The genes identified are related to cell growth, adhesion, apoptosis, signal transduction and bone formation. Furthermore, iSubpathwayMiner analyses were performed to identify significant biological pathway regions (subpathway) associated with this disease. The pathway analysis identified 11 statistically significant enriched subpathways, including pathways in cancer, p53 signaling pathway, osteoclast differentiation pathway and Wnt signaling pathway. Among these subpathways, four genes (IGF1, MDM2, STAT1 and RAC1) were presumed to play an important role in bone GCT recurrence. The differentially expressed MDM2 protein was immunohistochemically confirmed in the recurrent versus primary bone GCT tissues. This study identified differentially expressed genes and their subpathways in recurrent GCT, which may serve as potential biomarkers for the prediction of GCT recurrence. PMID:24969034

Chen, Shuxin; Li, Chunquan; Wu, Bingli; Zhang, Chunlong; Liu, Cheng; Lin, Xiaoxu; Wu, Xiangqiao; Sun, Lingling; Liu, Chunpeng; Chen, Bo; Zhong, Zhigang; Xu, Liyan; Li, Enmin

2014-09-01

119

Sodium current properties of primary skeletal myocytes and cardiomyocytes derived from different mouse strains  

PubMed Central

The mouse has become the preferred animal for genetic manipulations. Because of the diverse genetic backgrounds of various mouse strains, these can manifest strikingly different characteristics. Here, we studied the functional properties of currents through voltage-gated sodium channels in primary cultures of skeletal myocytes and cardiomyocytes derived from the three commonly used mouse strains BL6, 129/Sv, and FVB, by using the whole-cell patch-clamp technique. We found strain-specific sodium current function in skeletal myocytes, which could partly be explained by differences in sodium channel isoform expression. In addition, we found significant effects of cell source (neonatal or adult animal-derived) and variation of the differentiation time period. In contrast to skeletal myocytes, sodium current function in cardiomyocytes was similar in all strains. Our findings are relevant for the design and proper interpretation of electrophysiological studies, which use excitable cells in primary culture as a model system.

Mille, M.; Koenig, X.; Zebedin, E.; Uhrin, P.; Cervenka, R.; Todt, H.; Hilber, K.

2010-01-01

120

Depth dose dependence of the mouse bone using kilovoltage photon beams: A Monte Carlo study for small-animal irradiation  

NASA Astrophysics Data System (ADS)

This study investigated the dose enhancement due to the presence of mouse bone irradiated by the kilovoltage (kV) photon beams. Dosimetry of the bone associated with soft and lung tissue was determined by Monte Carlo simulations using the EGSnrc-based code in millimeter scale. Two inhomogeneous phantoms with 2 mm of bone layer sandwiched by: (1) water and lung (bone-lung phantom); and (2) water (bone-water phantom), were used. Relative depth doses along the central beam axes in the phantoms and dose enhancement ratios (bone dose in the above inhomogeneous phantoms to the dose at the same point in the water phantom) were determined using the 100 and 225 kVp photon beams. For the 100 kVp photon beams, the depth dose gradient in the bone was significantly larger compared to that in a water phantom without the bone. This is due to the beam hardening effect that some low-energy photons were filtered out in the deeper depth, resulting in less photoelectric interactions and hence energy depositions in the bone. Moreover, dose differences between the top and downstream (bottom) bone edges at depths of 1-5 mm were 168-192% and 149-166% for the bone-lung and bone-water phantom, respectively. These differences were larger than 21-27% (bone-lung) and 12-23% (bone-water) for the 225 kVp photon beams. The maximum dose enhancement ratio in the bone for the bone-lung and bone-water phantoms in various depths was about 5.7 using the 100 kVp photon beams. This ratio was larger than two times of that (2.4) for the 225 kVp photon beams. It is concluded that, apart from the basic beam characteristics such as attenuation and penumbra, which are related to the photon beam energy in the mouse irradiation, the bone dose is another important factor to consider when selecting the beam energy in the small-animal treatment planning, provided that the bone dose enhancement is a concern in the preclinical model.

Chow, James C. L.

2010-05-01

121

"In-bone" utricle cultures - A simplified, atraumatic technique for in situ cultures of the adult mouse (Mus musculus) utricle  

PubMed Central

Hypothesis The “in-bone” method of culturing utricles described here is a reliable and atraumatic technique for culturing mature mouse hair cells and studying hair cell death and protection. Background The current in vitro technique for studying hair cells of the mature mouse utricle involves removal from the temporal bone and free floating culture in media. This technique can be problematic due to variability in the preservation of the sensory epithelium and a steep learning curve that results in injury of the sensory epithelium in less experienced hands. We present a new atraumatic technique of culturing the utricle in situ within the temporal bone. Methods Leaving the temporal bone largely intact, a window is opened in the bony vestibule overlying the mouse utricle. The entire temporal bone is then placed into culture media. Utricles were cultured in situ for several days with minimal damage to the epithelium. The utricles are then fixed in situ, removed from the temporal bone, and processed. A standardized aminoglycoside-induced hair cell damage protocol was developed. Results Mature mouse utricles maintained hair cell numbers for 3 days in culture. Exposure to neomycin resulted in significant dose-dependent hair cell toxicity (p<.0001, one-way ANOVA). Exposure to the protective drug tacrine resulted in significant protection against neomycin (p<.05, three-way ANOVA). Conclusion The “in-bone” technique is a reliable and atraumatic method for culturing mature mouse utricles and studying hair cell death and protection. It is easily mastered and can make in vitro study of hair cells accessible to more research groups.

Ou, Henry C.; Lin, Vincent; Rubel, Edwin W

2013-01-01

122

Fusobacterium nucleatum and Tannerella forsythia Induce Synergistic Alveolar Bone Loss in a Mouse Periodontitis Model  

PubMed Central

Tannerella forsythia is strongly associated with chronic periodontitis, an inflammatory disease of the tooth-supporting tissues, leading to tooth loss. Fusobacterium nucleatum, an opportunistic pathogen, is thought to promote dental plaque formation by serving as a bridge bacterium between early- and late-colonizing species of the oral cavity. Previous studies have shown that F. nucleatum species synergize with T. forsythia during biofilm formation and pathogenesis. In the present study, we showed that coinfection of F. nucleatum and T. forsythia is more potent than infection with either species alone in inducing NF-?B activity and proinflammatory cytokine secretion in monocytic cells and primary murine macrophages. Moreover, in a murine model of periodontitis, mixed infection with the two species induces synergistic alveolar bone loss, characterized by bone loss which is greater than the additive alveolar bone losses induced by each species alone. Further, in comparison to the single-species infection, mixed infection caused significantly increased inflammatory cell infiltration in the gingivae and osteoclastic activity in the jaw bones. These data show that F. nucleatum subspecies and T. forsythia synergistically stimulate the host immune response and induce alveolar bone loss in a murine experimental periodontitis model.

Settem, Rajendra P.; El-Hassan, Ahmed Taher; Honma, Kiyonobu; Stafford, Graham P.

2012-01-01

123

Chondrogenic differentiation of mouse bone marrow mesenchymal stem cells induced by cartilage-derived morphogenetic protein-2 in vitro  

Microsoft Academic Search

Summary  To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2\\u000a in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human\\u000a cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng\\/mL). After 14 days of

Hongtao Tian; Shuhua Yang; Liang Xu; Yukun Zhang; Weihua Xu

2007-01-01

124

Gene Expression of Noncollagenous Bone Matrix Proteins in the Limb Joints and Intervertebral Disks of the twy Mouse  

Microsoft Academic Search

.   The twy (tiptoe walking Yoshimura) mouse is an autosomal recessive mutant manifesting multiple osteochondral lesions characterized\\u000a by pathologic calcium deposition. To elucidate the pathophysiology of the limb joint lesions and the intervertebral disk lesions\\u000a of the twy mouse, we assessed the mRNA expression of noncollagenous bone matrix proteins such as osteocalcin, osteonectin,\\u000a osteopontin, and matrix Gla protein (MGP) by

T. Ohtsuki; S. Furuya; T. Yamada; S. Nomura; J.-i. Hata; Y. Yabe; Y. Hosoda

1998-01-01

125

Comparison of bone tissue properties in mouse models with collagenous and non-collagenous genetic mutations using FTIRI.  

PubMed

Understanding how the material properties of bone tissue from the various forms of osteogenesis imperfecta (OI) differ will allow us to tailor treatment regimens for affected patients. To this end, we characterized the bone structure and material properties of two mouse models of OI, the osteogenesis imperfecta mouse (oim/oim) and fragilitas ossium (fro/fro), in which bone fragility is due to a genetic defect in collagen type I and a defect in osteoblast matrix mineralization, respectively. Bones from 3 to 6 month old animals were examined using Fourier transform infrared spectroscopic imaging (FTIRI), microcomputed tomography (micro-CT), histology, and biochemical analysis. The attributes of oim/oim bone tissue were relatively constant over time when compared to wild type animals. The mineral density in oim/oim cortices and trabecular bone was higher than wild type while the bones had thinner cortices and fewer trabeculae that were thinner and more widely spaced. The fro/fro animals exhibited osteopenic attributes at 3 months. However, by 6 months, their spectroscopic and geometric properties were similar to wild type animals. Despite the lack of a specific collagen defect in fro/fro mice, both fro/fro and oim/oim genotypes exhibited abnormal collagen crosslinking as determined by FTIRI at both time points. These results demonstrate that abnormal extracellular matrix assembly plays a role in the bone fragility in both of these models. PMID:22910579

Coleman, Rhima M; Aguilera, Laura; Quinones, Layla; Lukashova, Lyudamila; Poirier, Christophe; Boskey, Adele

2012-11-01

126

Comparison of bone tissue properties in mouse models with collagenous and non-collagenous genetic mutations using FTIRI  

PubMed Central

Understanding how the material properties of bone tissue from the various forms of osteogenesis imperfecta (OI) differ will allow us to tailor treatment regimens for affected patients. To this end, we characterized the bone structure and material properties of two mouse models of OI, the osteogenesis imperfecta mouse (oim/oim) and fragilitas ossium (fro/fro), in which bone fragility is due to a genetic defect in collagen type I and a defect in osteoblast matrix mineralization, respectively. Bones from 3 to 6 month old animals were examined using Fourier transform infrared spectroscopic imaging (FTIRI), microcomputed tomography (micro-CT), histology, and biochemical analysis. The attributes of oim/oim bone tissue were relatively constant over time when compared to wild type animals. The mineral density in oim/oim cortices and trabecular bone was higher than wild type while the bones had thinner cortices and fewer trabeculae that were thinner and more widely spaced. The fro/fro animals exhibited osteopenic attributes at 3 months. However, by 6 months, their spectroscopic and geometric properties were similar to wild type animals. Despite the lack of a specific collagen defect in fro/fro mice, both fro/fro and oim/oim genotypes exhibited abnormal collagen crosslinking as determined by FTIRI at both time points. These results demonstrate that abnormal extracellular matrix assembly plays a role in the bone fragility in both of these models.

Coleman, Rhima M.; Aguilera, Laura; Quinones, Layla; Lukashova, Lyudamila; Poirier, Christophe; Boskey, Adele

2012-01-01

127

Site-specific bone loss in senescence-accelerated mouse (SAMP6): a murine model for senile osteoporosis.  

PubMed

The senescence-accelerated mouse strain P6 (SAMP6) is a model of senile osteoporosis, which possesses many features of senile osteoporosis in humans. So far, little is known about the systemic bone microstructural changes that occur at multiple skeletal sites. In this study, we therefore, investigated site (vertebra, femur and tibia) dependence of bone microstructure and bone mineral density (BMD) in SAMP6 and the normal control mouse (SAMR1) at 5 and 12months of age using quantitative micro computed tomography (micro-CT) and image analysis software. As compared with SAMR1, the most prominent change in SAMP6 was the reduction of vertebral trabecular bone volume fraction (BV/TV) and trabecular BMD. Moderate decrease of trabecular bone mass was observed in the proximal tibia and distal femur. Increased marrow area and periosteal perimeter were investigated, though the cortical area and cortical thickness had no marked changes in the mid-tibial and mid-femoral cortical bones. These results indicate that bone microstructural properties in SAMP6 are remarkably heterogeneous throughout the skeleton, which is analogous to changes that occur in human bones. These findings further validate the relevance of SAMP6 as a model of senile osteoporosis. PMID:19815059

Chen, Huayue; Zhou, Xiangrong; Emura, Shoichi; Shoumura, Shizuko

2009-12-01

128

Direct observation of the major components of mouse bones and related compounds by electron Rutherford backscattering spectroscopy  

NASA Astrophysics Data System (ADS)

We present measurements, using electron Rutherford backscattering spectroscopy (eRBS), aiming to estimate the concentration of the various elements in mouse bone. We first successfully determined the composition of calcium carbonate, followed by an analysis of the more complicated case of hydroxyapatite. Finally we studied bone powder itself and established in this way that eRBS presents an interesting new flavor of microanalysis.

Vos, M.; Tökési, K.; Benkö, I.

2014-04-01

129

Stromal derived factor-1 regulates bone morphogenetic protein 2-induced osteogenic differentiation of primary mesenchymal stem cells  

PubMed Central

Stromal derived factor-1 (SDF-1) is a chemokine signaling molecule that binds to its transmembrane receptor CXC chemokine receptor-4 (CXCR4). While we previously detected that SDF-1 was co-required with bone morphogenetic protein 2 (BMP2) for differentiating mesenchymal C2C12 cells into osteoblastic cells, it is unknown whether SDF-1 is similarly involved in the osteogenic differentiation of mesenchymal stem cells (MSCs). Therefore, here we examined the role of SDF-1 signaling during BMP2-induced osteogenic differentiation of primary MSCs that were derived from human and mouse bone marrow. Our data showed that blocking of the SDF-1/CXCR4 signal axis or adding SDF-1 protein to MSCs significantly affected BMP2-induced alkaline phosphatase (ALP) activity and osteocalcin (OCN) synthesis, markers of preosteoblasts and mature osteoblasts, respectively. Moreover, disrupting the SDF-1 signaling impaired bone nodule mineralization during terminal differentiation of MSCs. Furthermore, we detected that blocking of the SDF-1 signaling inhibited the BMP2-induced early expression of Runt-related factor-2 (Runx2) and osterix (Osx), two “master” regulators of osteogenesis, and the SDF-1 effect was mediated via intracellular Smad and Erk activation. In conclusion, our results demonstrated a regulatory role of SDF-1 in BMP2-induced osteogenic differentiation of MSCs, as perturbing the SDF-1 signaling affected the differentiation of MSCs towards osteoblastic cells in response to BMP2 stimulation. These data provide novel insights into molecular mechanisms underlying MSC osteogenesis, and will contribute to the development of MSC therapies for enhancing bone formation and regeneration in broad orthopaedic situations.

Hosogane, Naobumi; Huang, Zhiping; Rawlins, Bernard A.; Liu, Xia; Boachie-Adjei, Oheneba; Boskey, Adele L.; Zhu, Wei

2010-01-01

130

Induction of bone formation in biphasic calcium phosphate scaffolds by bone morphogenetic protein-2 and primary osteoblasts.  

PubMed

Bone tissue engineering strategies mainly depend on porous scaffold materials. In this study, novel biphasic calcium phosphate (BCP) matrices were generated by 3D-printing. High porosity was achieved by starch consolidation. This study aimed to characterise the porous BCP-scaffold properties and interactions of osteogenic cells and growth factors under in vivo conditions. Five differently treated constructs were implanted subcutaneously in syngeneic rats: plain BCP constructs (group A), constructs pre-treated with BMP-2 (group B; 1.6?µg BMP-2 per scaffold), seeded with primary osteoblasts (OB) (group C), seeded with OB and BMP-2 (group D) and constructs seeded with OB and pre-cultivated in a flow bioreactor for 6?weeks (group E). After 2, 4 and 6?weeks, specimens were explanted and subjected to histological and molecular biological analyses. Explanted scaffolds were invaded by fibrovascular tissue without significant foreign body reactions. Morphometric analysis demonstrated significantly increased bone formation in samples from group D (OB?+?BMP-2) compared to all other groups. Samples from groups B-E displayed significant mRNA expression of bone-specific genes after 6?weeks. Pre-cultivation in the flow bioreactor (group E) induced bone formation comparable with group B. In this study, differences in bone distribution between samples with BMP-2 or osteoblasts could be observed. In conclusion, combination of osteoblasts and BMP-2 synergistically enhanced bone formation in novel ceramic scaffolds. These results provide the basis for further experiments in orthotopic defect models with a focus on future applications in orthopaedic and reconstructive surgery. PMID:22740314

Strobel, L A; Rath, S N; Maier, A K; Beier, J P; Arkudas, A; Greil, P; Horch, R E; Kneser, U

2014-03-01

131

Multimodality Imaging of Tumor and Bone Response in a Mouse Model of Bony Metastasis1  

PubMed Central

Cancer drug development generally performs in vivo evaluation of treatment effects that have traditionally relied on detection of morphologic changes. The emergence of new targeted therapies, which may not result in gross morphologic changes, has spurred investigation into more specific imaging methods to quantify response, such as targeted fluorescent probes and bioluminescent cells. The present study investigated tissue response to docetaxel or zoledronic acid (ZA) in a mouse model of bony metastasis. Intratibial implantations of breast cancer cells (MDA-MB-231) were monitored throughout this study using several modalities: molecular resonance imaging (MRI) tumor volume and apparent diffusion coefficient (ADC), micro-computed tomography (µCT) bone volume, bioluminescence imaging (BLI) reporting cancer cell apoptosis, and fluorescence using Osteosense 800 and CatK 680-FAST. Docetaxel treatment resulted in tumor cell kill reflected by ADC and BLI increases and tumor volume reduction, with delayed bone recovery seen in µCT prefaced by increased osteoblastic activity (Osteosense 800). In contrast, the ZA treatment group produced similar values in MRI, BLI, and Osteosense 800 fluorescence imaging readouts when compared to controls. However, µCT bone volume increased significantly by the first week post-treatment and the CatK 680-FAST signal was slightly diminished by 4 weeks following ZA treatment. Multimodality imaging provides a more comprehensive tool for new drug evaluation and efficacy screening through identification of morphology as well as function and apoptotic signaling.

Hoff, Benjamin A; Chughtai, Komal; Jeon, Yong Hyun; Kozloff, Kenneth; Galban, Stefanie; Rehemtulla, Alnawaz; Ross, Brian D; Galban, Craig J

2012-01-01

132

Impaired cholesterol esterification in primary brain cultures of the lysosomal cholesterol storage disorder (LCSD) mouse mutant  

SciTech Connect

Esterification of cholesterol was investigated in primary neuroglial cultures obtained from newborn lysosomal cholesterol storage disorder (LCSD) mouse mutants. An impairment in /sup 3/H-oleic acid incorporation into cholesteryl esters was demonstrated in cultures of homozygous LCSD brain. Primary cultures derived from other phenotypically normal pups of the carrier breeders esterified cholesterol at normal levels or at levels which were intermediary between normal and deficient indicating a phenotypic expression of the LCSD heterozygote genotype. These observations on LCSD mutant brain cells indicate that the defect in cholesterol esterification is closely related to the primary genetic defect and is expressed in neuroglial cells in culture.

Patel, S.C.; Suresh, S.; Weintroub, H.; Brady, R.O.; Pentchev, P.G.

1987-02-27

133

Improved quantitative analysis of primary bone marrow megakaryocytes utilizing imaging flow cytometry.  

PubMed

Life-threatening thrombocytopenia can develop following bone marrow injury due to decreased platelet production from megakaryocytes (MKs). However, the study of primary MKs has been complicated by their low frequency in the bone marrow and by technical challenges presented by their unique maturation properties. More accurate and efficient methods for the analysis of in vivo MKs are needed to enhance our understanding of megakaryopoiesis and ultimately develop new therapeutic strategies for thrombocytopenia. Imaging flow cytometry (IFC) combines the morphometric capabilities of microscopy with the high-throughput analyses of flow cytometry (FC). Here, we investigate the application of IFC on the ImageStream(X) platform to the analysis of primary MKs isolated from murine bone marrow. Our data highlight and address technical challenges for conventional FC posed by the wide range of cellular size within the MK lineage as well as the shared surface phenotype with abundant platelet progeny. We further demonstrate that IFC can be used to reproducibly and efficiently quantify the frequency of primary murine MKs in the marrow, both at steady-state and in the setting of radiation-induced bone marrow injury, as well as assess their ploidy distribution. The ability to accurately analyze the full spectrum of maturing MKs in the bone marrow now allows for many possible applications of IFC to enhance our understanding of megakaryopoiesis and platelet production. PMID:24616422

Niswander, Lisa M; McGrath, Kathleen E; Kennedy, John C; Palis, James

2014-04-01

134

Validated Laser Doppler protocol for measurement of mouse bone blood perfusion - response to age or ovariectomy differs with genetic background.  

PubMed

The physiological role of bone vascularization in bone metabolism begins to be understood; however, its involvement in pathological situations remains poorly explored. Bone blood supply depends on both vascular density and blood flow. However, in mice, the specific evaluation of perfusion in bone suffers from a lack of easy-handling measurement tools. In the present study, we first developed a Laser Doppler Perfusion Measurement (LDPM) protocol in mouse tibia, which we validated with ex vivo and in vivo experiments. Then we carried out a study associating both structural (vascular quantitative histomorphometry) and functional (LDPM) approaches. We studied the effects of aging in 4, 7 and 17 month-old male mice and the early effects of ovariectomy in 4 month-old females. Both studies were carried out in inbred mice (C57BL/6) and in mice of mixed background (129sv/CD1). The significant differences we observed between strains in unchallenged 4 month-old animals concerned both perfusion and vascular density and depended on gender. Additionally, the age-related bone loss observed in male mice was not temporally associated with vascular changes in either strain. Between 7 and 17 months, we did not find any decrease in bone vascular density or perfusion. In contrast, ovariectomy triggered early vascular structural and functional adaptations which differed between genetic backgrounds. We observed that bone vessel density did not generally account for bone perfusion levels. In conclusion, we describe here a LDPM-based experimental protocol which provides a reproducible quantitative evaluation of bone perfusion in mouse tibia, hence allowing intergroup comparisons. This integrative structural and functional approach of bone vascularization showed that bone vascular adaptation occurs during aging or after ovariectomy and is affected by the genetic background. PMID:23571049

Roche, Bernard; Vanden-Bossche, Arnaud; Normand, Myriam; Malaval, Luc; Vico, Laurence; Lafage-Proust, Marie-Hélène

2013-08-01

135

Primary B-lymphoblastic lymphoma of gallbladder involving mandibular bone.  

PubMed

We report the case of a 75-year-old man who presented for evaluation of painless hematuria persisting for more than 1 month. At the time of presentation, the patient did not report any systemic symptoms and had no fever, weight loss, or dysuria. Computed tomography showed several enhancing, sessile polyps in the gall bladder (1.5 cm or smaller). There was no associated stone or biliary dilation. Since no other abnormality was evident, we performed laparoscopic cholecystectomy. He was diagnosed as having B-cell lymphoblastic lymphoma (B-LBL) after surgical resection of the gall bladder (GB). As the left mandibular swelling was developed after the diagnosis of the B-LBL involving GB, facial magnetic resonance imaging (MRI) was added to the imaging scan. Facial MRI revealed mass formation in the left mandible, left medial pterygoid, masticator, and buccinator muscles. The biopsy samples from the mandibular bone were also diagnosed as B-LBL. The definitive pathological diagnosis was B-LBL, stage IV. Systemic chemotherapy was done with subsequent response in size of the left mandible mass. PMID:24789124

Kim, Hee-Jun; Lee, Tae Jin; Choi, Yoo Shin

2014-06-01

136

Generation of competent bone marrow-derived antigen presenting cells from the deer mouse (Peromyscus maniculatus)  

PubMed Central

Background Human infections with Sin Nombre virus (SNV) and related New World hantaviruses often lead to hantavirus cardiopulmonary syndrome (HCPS), a sometimes fatal illness. Lungs of patients who die from HCPS exhibit cytokine-producing mononuclear infiltrates and pronounced pulmonary inflammation. Deer mice (Peromyscus maniculatus) are the principal natural hosts of SNV, in which the virus establishes life-long persistence without conspicuous pathology. Little is known about the mechanisms SNV employs to evade the immune response of deer mice, and experimental examination of this question has been difficult because of a lack of methodologies for examining such responses during infection. One such deficiency is our inability to characterize T cell responses because susceptible syngeneic deer mice are not available. Results To solve this problem, we have developed an in vitro method of expanding and generating competent antigen presenting cells (APC) from deer mouse bone marrow using commercially-available house mouse (Mus musculus) granulocyte-macrophage colony stimulating factor. These cells are capable of processing and presenting soluble protein to antigen-specific autologous helper T cells in vitro. Inclusion of antigen-specific deer mouse antibody augments T cell stimulation, presumably through Fc receptor-mediated endocytosis. Conclusions The use of these APC has allowed us to dramatically expand deer mouse helper T cells in culture and should permit extensive characterization of T cell epitopes. Considering the evolutionary divergence between deer mice and house mice, it is probable that this method will be useful to other investigators using unconventional models of rodent-borne diseases.

Davenport, Bennett J; Willis, Derall G; Prescott, Joseph; Farrell, Regina M; Coons, Teresa A; Schountz, Tony

2004-01-01

137

Cortical and trabecular bone adaptation to incremental load magnitudes using the mouse tibial axial compression loading model.  

PubMed

The mouse tibial axial compression loading model has recently been described to allow simultaneous exploration of cortical and trabecular bone adaptation within the same loaded element. However, the model frequently induces cortical woven bone formation and has produced inconsistent results with regards to trabecular bone adaptation. The aim of this study was to investigate bone adaptation to incremental load magnitudes using the mouse tibial axial compression loading model, with the ultimate goal of revealing a load that simultaneously induced lamellar cortical and trabecular bone adaptation. Adult (16 weeks old) female C57BL/6 mice were randomly divided into three load magnitude groups (5, 7 and 9N), and had their right tibia axially loaded using a continuous 2-Hz haversine waveform for 360 cycles/day, 3 days/week for 4 consecutive weeks. In vivo peripheral quantitative computed tomography was used to longitudinally assess midshaft tibia cortical bone adaptation, while ex vivo micro-computed tomography and histomorphometry were used to assess both midshaft tibia cortical and proximal tibia trabecular bone adaptation. A dose response to loading magnitude was observed within cortical bone, with increasing load magnitude inducing increasing levels of lamellar cortical bone adaptation within the upper two thirds of the tibial diaphysis. Greatest cortical bone adaptation was observed at the midshaft where there was a 42% increase in estimated mechanical properties (polar moment of inertia) in the highest (9N) load group. A dose response to load magnitude was not clearly evident within trabecular bone, with only the highest load (9N) being able to induce measureable adaptation (31% increase in trabecular bone volume fraction at the proximal tibia). The ultimate finding was that a load of 9N (engendering a tensile strain of 1833 ?? on medial surface of the midshaft tibia) was able to simultaneously induce measurable lamellar cortical and trabecular bone adaptation when using the mouse tibial axial compression loading model in 16 week old female C57BL/6 mice. This finding will help plan future studies aimed at exploring simultaneous lamellar cortical and trabecular bone adaptation within the same loaded element. PMID:23111313

Weatherholt, Alyssa M; Fuchs, Robyn K; Warden, Stuart J

2013-01-01

138

Cortical and Trabecular Bone Adaptation to Incremental Load Magnitudes Using the Mouse Tibial Axial Compression Loading Model  

PubMed Central

The mouse tibial axial compression loading model has recently been described to allow simultaneous exploration of cortical and trabecular bone adaptation within the same loaded element. However, the model frequently induces cortical woven bone formation and has produced inconsistent results with regards to trabecular bone adaptation. The aim of this study was to investigate bone adaptation to incremental load magnitudes using the mouse tibial axial compression loading model, with the ultimate goal of revealing a load that simultaneously induced lamellar cortical and trabecular bone adaptation. Adult (16 week old) female C57BL/6 mice were randomly divided into three load magnitude groups (5, 7 and 9 N), and had their right tibia axially loaded using a continuous 2-Hz haversine waveform for 360 cycles/d, 3 d/wk for 4 consecutive weeks. In vivo peripheral quantitative computed tomography was used to longitudinally assess midshaft tibia cortical bone adaptation, while ex vivo micro-computed tomography and histomorphometry were used to assess both midshaft tibia cortical and proximal tibia trabecular bone adaptation. A dose response to loading magnitude was observed within cortical bone, with increasing load magnitude inducing increasing levels of lamellar cortical bone adaptation within the upper two thirds of the tibial diaphysis. Greatest cortical bone adaptation was observed at the midshaft where there was a 42% increase in estimated mechanical properties (polar moment of inertia) in the highest (9 N) load group. A dose response to load magnitude was not clearly evident within trabecular bone, with only the highest load (9 N) being able to induce measureable adaptation (31% increase in trabecular bone volume fraction at the proximal tibia). The ultimate finding was that a load of 9 N (engendering a tensile strain of 1,833 ?? on medial surface of the midshaft tibia) was able to simultaneously induce measurable lamellar cortical and trabecular bone adaptation when using the mouse tibial axial compression loading model in 16 week old female C57BL/6 mice. This finding will help plan future studies aimed at exploring simultaneous lamellar cortical and trabecular bone adaptation within the same loaded element.

Weatherholt, Alyssa M.; Fuchs, Robyn K.; Warden, Stuart J.

2012-01-01

139

[Isolation of mesenchymal stem cells from bone marrow filters by primary explant culture].  

PubMed

This study was aimed to investigate whether mesenchymal stem cells (MSC) can be isolated from bone marrow filters which have always been discarded. The bone marrow (BM) particles from BM filters of 2 healthy donors were cultivated by primary explant culture. After expansion, the number of MSC was counted and their immunophenotype and differentiation potential were detected. The results indicated that many MSC were found in bone marrow particles from filters, and nearly 10(7) MSC were obtained at 3 passages of expansion. They not only possessed the characteristics of morphology and immunophenotype of MSC, but also could differentiate into osteoblasts, chondrocytes and adipocytes. In conclusion, a large amount of MSC can be obtained from BM filters if the BM particles were cultivated by primary explant culture. PMID:21518508

Xing, Wen; Yang, Shao-Guang; Liu, Meng; Lu, Shi-Hong; Zhao, Qin-Jun; Pang, Ai-Ming; Yao, Jian-Feng; Li, Jian-Ping; Ren, Qian; Han, Zhong-Chao

2011-04-01

140

Percutaneous Ethibloc injection in the treatment of primary aneurysmal bone cysts.  

PubMed

The effects of percutaneous Ethibloc (Ethicon/Johnson & Johnson, St-Stevens-Woluwe, Belgium) injection into primary aneurysmal bone cysts were analysed. Two patients with a venous drainage after injection of a medium contrast were excluded. Twelve patients underwent at least one percutaneous injection of Ethibloc. The average follow-up period was 5.1 years. At final follow-up, six patients had complete healing of the cyst, three had partial healing and three, who had no response, were treated by curettage and bone grafting. Complete healing was observed for all the aggressive lesions. No major complications were noted. Ethibloc injection may be performed as a primary treatment of aneurysmal bone cysts if the technique is followed with precision. PMID:16093949

de Gauzy, Jérôme Sales; Abid, Abdelazis; Accadbled, Franck; Knorr, Gorka; Darodes, Philippe; Cahuzac, Jean Philippe

2005-09-01

141

Knee loading stimulates healing of mouse bone wounds in a femur neck  

PubMed Central

Healing of bone wounds is sensitive to various environmental stimuli. Using knee loading, which has been shown to stimulate bone formation in mouse femora and tibiae, we addressed a question: Does knee loading accelerate a closure of open wounds in a femur neck? A surgical wound (0.5 mm in diameter) was generated at the femur neck in the left and right femora of C57/BL/6 female mice, and knee loading was applied to the left knee for 3 min/day for 3 consecutive days. Surgical holes at the femoral midshaft were used as control. Animals were sacrificed 1, 2, and 3 weeks after surgery for analyses with µCT and pQCT as well as mechanical testing. The results showed load-driven acceleration of the closure of surgical holes. Compared to a sham-loaded contralateral control, knee loading reduced the size of surgical wounds in the femoral midshaft by 14% (p < 0.05), 21% (p < 0.01), and 32% (p < 0.001) in 1, 2, and 3 weeks, respectively. It also decreased the wound size in the femur neck by 16% (p < 0.001; 1 week), 18% (p < 0.001; 2 weeks), and 21% (p < 0.001; 3 weeks). Images with pQCT revealed that bone mineral density (BMD) was increased from 571 ± 19 mg/cm3 (control) to 686 ± 19 mg/cm3 (loaded) (p < 0.01), and bone mineral content (BMC) from 3.05 ± 0.12 mg/mm (control) to 3.42 ± 0.11 mg/mm (loaded) (p < 0.05). Furthermore, mechanical testing showed that stiffness of the femur was increased by knee loading (p < 0.05). This study demonstrates that knee loading is capable of accelerating healing of surgical wounds throughout the femur including the femoral midshaft and neck.

Zhang, Ping; Yokota, Hiroki

2011-01-01

142

Tripterygium hypoglaucum (level) Hutch induces aneuploidy of chromosome 8 in mouse bone marrow cells and sperm.  

PubMed

Aneuploidy of mouse chromosome 8 induced by a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH) was investigated by fluorescence in situ hybridization (FISH) in vivo. Male mice were treated with THH (single i.p. injection) at doses of 120, 240 and 480 mg/kg. Colchicine (COL, 1.5 mg/kg i.p.) was used as a positive control. Bone marrow cells and epididymal sperm were collected 24 h and 22 days after treatment, respectively. Chromosome 8 aneuploidies induced by THH in bone marrow cells and sperm were determined by FISH with a biotin-16-dUTP labelled DNA probe corresponding to the centromeric region of chromosome 8. The hybridized probe was detected with avidin-FITC. The frequencies of trisomy 8 in bone marrow cells were 0.16% in the solvent control group, 0.39% in the COL-treated group and 0.33, 0.41 and 0.41% in the THH-treated groups, respectively. The frequencies of disomy 8 sperm were 0.11% in the solvent control group, 0.27% in the COL-treated group and 0.23, 0.27 and 0.27% in the THH-treated groups, respectively. The experiment showed that induced aneuploidy frequencies were higher in bone marrow cells than in sperm with COL and the two higher doses of THH (P < 0.05). All groups were significantly different from the corresponding solvent controls (P < 0.01-0.001), but there was no dose-related increase in either cell type. Considering the present results together with our previous studies, it appears that THH is a potent mammalian aneugen which may pose a genetic risk to human patients. PMID:15388810

Xu, Wang; Ziqing, Liang; Yinrun, Ding; Xiaoyan, Wang; Jinglun, Xue

2004-09-01

143

Bone conducted vibration selectively activates irregular primary otolithic vestibular neurons in the guinea pig  

Microsoft Academic Search

The main objective of this study was to determine whether bone-conducted vibration (BCV) is equally effective in activating both semicircular canal and otolith afferents in the guinea pig or whether there is preferential activation of one of these classes of vestibular afferents. To answer this question a large number (346) of single primary vestibular neurons were recorded extracellularly in anesthetized

Ian S. Curthoys; Juno Kim; Samara K. McPhedran; Aaron J. Camp

2006-01-01

144

Radiation sensitivity and cycling status of mouse bone marrow prothymocytes and day 8 colony forming units spleen (CFUs)  

SciTech Connect

Mouse bone marrow prothymocytes as determined in an in vivo thymus regeneration assay have an in vitro gamma radiation sensitivity which is different from that of spleen colony forming cells (CFUs). Determination of Do according to in vivo irradiation revealed similar but insignificant differences. Prothymocytes in normal bone marrow maintain a low but slightly different proliferative state as compared to CFUs, according to determinations using the /sup 3/H-TdR suicide technique. In regenerating bone marrow prothymocytes were found to be sensitive to an inhibitory effect of in vitro incubation with cold thymidine. CFUs and normal bone marrow prothymocytes were not affected by cold thymidine. Taking into account the cold thymidine effect it can be concluded that prothymocytes and CFUs in regenerating bone marrow are fully in cycle. These results are best explained when prothymocytes and CFUs are considered to be different cells.

Boersma, W.J.

1983-11-01

145

Cell Fusion Reprogramming Leads to a Specific Hepatic Expression Pattern during Mouse Bone Marrow Derived Hepatocyte Formation In Vivo  

Microsoft Academic Search

The fusion of bone marrow (BM) hematopoietic cells with hepatocytes to generate BM derived hepatocytes (BMDH) is a natural process, which is enhanced in damaged tissues. However, the reprogramming needed to generate BMDH and the identity of the resultant cells is essentially unknown. In a mouse model of chronic liver damage, here we identify a modification in the chromatin structure

Oscar Quintana-Bustamante; Esther Grueso; Ramon Garcia-Escudero; Elvira Arza; Alberto Alvarez-Barrientos; Isabel Fabregat; Maria Garcia-Bravo; Nestor W. Meza; Jose C. Segovia

2012-01-01

146

Prognostic relevance of disseminated tumor cells in the bone marrow and biological factors of 265 primary breast carcinomas  

Microsoft Academic Search

INTRODUCTION: The prognostic significance of disseminated tumor cells in the bone marrow (DTC-BM) of breast cancer patients has been demonstrated in many studies. Yet, it is not clear which of the primary tumors' biological factors predict hematogenous dissemination. We therefore examined 'tissue micro arrays' (TMAs) of 265 primary breast carcinomas from patients with known bone marrow (BM) status for HER2,

Christian Schindlbeck; Theresa Kampik; Wolfgang Janni; Brigitte Rack; Udo Jeschke; Stan Krajewski; Harald Sommer; Klaus Friese

2005-01-01

147

A primary culture system of mouse thick ascending limb cells with preserved function and uromodulin processing.  

PubMed

The epithelial cells lining the thick ascending limb (TAL) of the loop of Henle perform essential transport processes and secrete uromodulin, the most abundant protein in normal urine. The lack of differentiated cell culture systems has hampered studies of TAL functions. Here, we report a method to generate differentiated primary cultures of TAL cells, developed from microdissected tubules obtained in mouse kidneys. The TAL tubules cultured on permeable filters formed polarized confluent monolayers in ?12 days. The TAL cells remain differentiated and express functional markers such as uromodulin, NKCC2, and ROMK at the apical membrane. Electrophysiological measurements on primary TAL monolayers showed a lumen-positive transepithelial potential (+9.4?±?0.8 mV/cm(2)) and transepithelial resistance similar to that recorded in vivo. The transepithelial potential is abolished by apical bumetanide and in primary cultures obtained from ROMK knockout mice. The processing, maturation and apical secretion of uromodulin by primary TAL cells is identical to that observed in vivo. The primary TAL cells respond appropriately to hypoxia, hypertonicity, and stimulation by desmopressin, and they can be transfected. The establishment of this primary culture system will allow the investigation of TAL cells obtained from genetically modified mouse models, providing a critical tool for understanding the role of that segment in health and disease. PMID:23887378

Glaudemans, Bob; Terryn, Sara; Gölz, Nadine; Brunati, Martina; Cattaneo, Angela; Bachi, Angela; Al-Qusairi, Lama; Ziegler, Urs; Staub, Olivier; Rampoldi, Luca; Devuyst, Olivier

2014-02-01

148

[Effect of osteogenically and adipogenically differentiated bone mesenchymal stem cells from mouse on osteoclast formation].  

PubMed

This study was purposed to investigate the regulatory effects of differentiating mesenchymal stem cells (MSC) on osteoclast formation. The MSC from mouse compact bones were cultured and induced into osteoblasts and adipocytes for one week. To test their regulatory effect on osteoclastogenesis, osteogenically differentiated and adipogenically differentiated MSC were co-cultured with CD11b(+) monocytes and osteoclasts were identified with in situ tartrate-resistant acid phosphatase (TRAP) staining. The results showed that differentiated MSC supported osteoclastogenesis but the osteoclast supporting capacity of osteogenically differentiated MSC decreased as compared with undifferentiated MSC. More interestingly, the adipogenically differentiated MSC significantly promoted osteoclasts formation when co-cultured with monocytes. It is concluded that the regulatory effect of MSC on osteoclast formation has changed while they have differentiated into different types of cells. The findings indicate that MSC may exert alternative effect on osteoclastogenesis by differentiation to descendant cells. PMID:23114145

Zhu, Heng; Liu, Yuan-Lin; Chen, Ji-De; Li, Hong; Liu, Yu-Xiao; Xu, Fen-Fen; Jiang, Xiao-Xia; Zhang, Yi; Mao, Ning

2012-10-01

149

Isolation of common dendritic cell progenitors (CDP) from mouse bone marrow.  

PubMed

In the steady-state lymphoid organ, dendritic cells (DCs) are classified into two major subsets, plasmacytoid DC (pDC) and conventional DC (cDC). A standing question was whether a common progenitor for plasmacytoid and conventional dendritic cells exists during the sequential differentiation from hematopoietic stem cells to dendritic cells. We have recently identified such a common clonogenic plasmacytoid and dendritic cell progenitor (CDP) from mouse bone marrow using antibodies for c-kit, Flt3, and M-CSFR. CDPs generated almost exclusively pDC and cDC in vitro and upon transfer in irradiated and steady-state mice in vivo. Single-cell analysis revealed the existence of clonal progenitors giving rise to both pDC and cDC within the CDP population. Thus, these results prove the existence of a common developmental pathway for at least some pDCs and cDCs in lymphoid organs in vivo. PMID:19941114

Onai, Nobuyuki; Manz, Markus G; Schmid, Michael A

2010-01-01

150

Primary bone lymphoplasmacytic lymphoma presenting with spinal cord compression: a case report.  

PubMed

Primary bone lymphoma is a rare disease, and the main pathological type is diffuse large B-cell lymphoma. The occurrence of follicular, marginal zone and lymphoplasmacytic lymphomas is rare. Vertebras are also sites that can be affected, and spinal cord compression is reported in 14% of patients with vertebral involvement. However, there is no report on primary vertebral lymphoplasmacytic lymphoma with spinal cord compression. The present report presents one case of primary vertebral lymphoplasmacytic lymphoma with spinal cord compression and increased serum and urine ? light chain, without an elevated heavy chain of immunoglobulin. Conflict of interest:None declared. PMID:24385833

Lei, Yang; Zi, Liu; Long, Su; Pei, Li; Wei, Li

2013-12-01

151

Cytogenetic effects of sildenafil citrate (Viagra) on SWR/J mouse bone marrow cells  

PubMed Central

The present study was conducted to investigate the cytogenetic effects of sildenafil citrate in SWR/J mouse bone marrow cells. Thirty-six males and 36 females were used and divided into four groups. Each group contained 18 animals (9 males and 9 females), weighing 30–35 g. These animals were orally administered with a single dose of 13, 26 or 40 mg/kg sildenafil citrate solution. A control group received normal saline in an identical condition. The animals were sacrificed at 12, 24 or 48 h, after the treatment. Chromosome aberrations were investigated in 50 metaphases per animal. No significant differences in the percentages of mitotic indices or in the frequencies of chromosome aberrations were observed between treated male and female mice at any doses or at any time intervals used, therefore, data from the two sexes were pooled when analyzed statistically. No significant (p < 0.05) differences in the percentages of mitotic indices or in the frequencies of chromosome aberrations were observed between sildenafil citrate-treated groups and the control group at any doses or at any time intervals used. However, the percentages of centromeric adhesions increased significantly (p < 0.01) in treated groups as compared with the control group at all doses and at all time intervals used. In conclusion, the results of the present study suggest that sildenafil citrate does not have cytogenetic effects on mouse bone marrow cells, but the centromeric adhesions induced by this drug need further studies to confirm them and to investigate the possible mechanism(s) responsible for such effect.

Abou-Tarboush, Faisal Mohamed; Abdel-Samad, Mohamed Fathy

2010-01-01

152

Cytogenetic effects of sildenafil citrate (Viagra) on SWR/J mouse bone marrow cells.  

PubMed

The present study was conducted to investigate the cytogenetic effects of sildenafil citrate in SWR/J mouse bone marrow cells. Thirty-six males and 36 females were used and divided into four groups. Each group contained 18 animals (9 males and 9 females), weighing 30-35 g. These animals were orally administered with a single dose of 13, 26 or 40 mg/kg sildenafil citrate solution. A control group received normal saline in an identical condition. The animals were sacrificed at 12, 24 or 48 h, after the treatment. Chromosome aberrations were investigated in 50 metaphases per animal. No significant differences in the percentages of mitotic indices or in the frequencies of chromosome aberrations were observed between treated male and female mice at any doses or at any time intervals used, therefore, data from the two sexes were pooled when analyzed statistically. No significant (p < 0.05) differences in the percentages of mitotic indices or in the frequencies of chromosome aberrations were observed between sildenafil citrate-treated groups and the control group at any doses or at any time intervals used. However, the percentages of centromeric adhesions increased significantly (p < 0.01) in treated groups as compared with the control group at all doses and at all time intervals used. In conclusion, the results of the present study suggest that sildenafil citrate does not have cytogenetic effects on mouse bone marrow cells, but the centromeric adhesions induced by this drug need further studies to confirm them and to investigate the possible mechanism(s) responsible for such effect. PMID:23961094

Abou-Tarboush, Faisal Mohamed; Abdel-Samad, Mohamed Fathy

2010-10-01

153

RAMAN AND MECHANICAL PROPERTIES CORRELATE AT WHOLE BONE- AND TISSUE- LEVELS IN A GENETIC MOUSE MODEL  

PubMed Central

The fracture resistance of bone arises from the composition, orientation, and distribution of the primary constituents at each hierarchical level of organization. Therefore, to establish the relevance of Raman Spectroscopy (RS) in identifying differences between strong or tough bone and weak or brittle bone, we investigated whether Raman-derived properties could explain the variance in biomechanical properties at both the whole bone and the tissue-level, and do so independently of traditional measurements of mineralization. We harvested femurs from wild-type mice and mice lacking matrix metalloproteinase 2 because the mutant mice have a known reduction in mineralization. Next, RS quantified compositional properties directly from the intact diaphysis followed by micro-computed tomography to quantify mineralization density (Ct.TMD). Correlations were then tested for significance between these properties and the biomechanical properties as determined by the three point bending test on the same femurs. Harvested tibia were embedded in plastic, sectioned transversely, and polished in order to acquire average Raman properties per specimen that were then correlated with average nanoindentation properties per specimen. Dividing the ?1 phosphate by the proline peak intensity provided the strongest correlation between the mineral-to-collagen ratio and the biomechanical properties (whole bone modulus, strength and post-yield deflection plus nanoindentation modulus). Moreover, the linear combination of ?1 phosphate/proline and Ct.TMD provided the best explanation of the variance in strength between the genotypes, and it alone was the best explanatory variable for brittleness. Causal relationships between Raman and fracture resistance need to be investigated, but Raman has the potential to assess fracture risk.

Bi, Xiaohong; Patil, Chetan A.; Lynch, Conor C.; Pharr, George M.; Mahadevan-Jansen, Anita; Nyman, Jeffry S.

2010-01-01

154

Bone Metastasis from Primary Hepatocellular Carcinoma: Characteristics of Soft Tissue Formation  

PubMed Central

Purpose To assess the characteristics of bone metastasis from hepatocellular carcinoma and the radiation field arrangement based on imaging studies. Materials and Methods Fifty-three patients (84 lesions) with bone metastasis from a primary hepatocellular carcinoma completed palliative radiation therapy. All patients underwent one of following imaging studies prior to the initiation of radiation therapy: a bone scan, computed tomography or magnetic resonance imaging. The median radiation dose was 30 Gy (7~40 Gy). We evaluated retrospectively the presence of soft tissue formation and the adjustment of the radiation field based on the imaging studies. Results Soft tissue formation at the site of bony disease was identified from either a CT/MRI scan (41 lesions) or from a symptomatic palpable mass (5 lesions). The adjustment of the radiation field size based on a bone scan was necessary for 31 of 41 soft tissue forming lesions (75.6%), after a review of the CT/MRI scan. The median survival from the initial indication of a hepatoma diagnosis was 8 months (2 to 71 months), with a 2-year survival rate of 38.6%. The median survival from the detection of a bone metastasis was 5 months (1 to 38 months) and the 1-year overall survival rate was 8.7%. Conclusion It was again identified that bone metastasis from a primary hepatocellular carcinoma is accompanied by soft tissue formation. From this finding, an adjustment of the radiation field size based on imaging studies is required. It is advisable to obtain a CT or MRI scan of suspected bone metastasis for better tumor volume coverage prior to the initiation of radiation therapy.

Kim, Sangwon; Wang, Heejung; Cho, Sungwon; Oh, Young-Taek; Kang, Seung-Hee; Yang, Juno

2007-01-01

155

Changes in trabecular bone, hematopoiesis and bone marrow vessels in aplastic anemia, primary osteoporosis, and old age: A comparative histomorphometric study  

Microsoft Academic Search

Retrospective histologic analyses of bone biopsies and of post mortem samples from normal persons of dif- ferent age groups, and of bone biopsies of age- and sex-matched groups of patients with primary osteo- porosis and aplastic anemia show characteristic age dependent as well as pathologic changes including at- rophy of osseous trabeculae and of hematopoiesis, and changes in the sinusoidal

G. Kettner; W. BijHM; M. Schmidmeier; R. Schlag; B. Frisch; B. Mallmann; W. Eisenmenger; Th. Gilg

1987-01-01

156

Multiple soft tissue aneurysmal cysts: An occurrence after resection of primary aneurysmal bone cyst of fibula  

PubMed Central

We report a case of multiple extraosseous aneurysmal cysts occurring in the muscle and subcutaneous plane of postero-lateral aspects of the upper right leg. They appeared about 15 months after resection of aneurysmal bone cyst of the upper end of the fibula. They varied in size from 2 cm to 5 cm. Radiologically they were well-defined lesions with central septate areas surrounded by a rim of calcification. Histologically they showed central cystic spaces separated by septa consisting of fibroblasts, osteoclast type of giant cells and reactive woven bone. Thus they showed histological similarity with aneurysmal bone cysts, but did not show any connection with the bone. Only very few examples of aneurysmal cysts of soft tissue had been described in the past one decade and they were reported in various locations including rare sites such as arterial wall and larynx. Recent cytogenetic analyses have shown abnormalities involving 17p11-13 and/or 16q22 in both osseous and extraosseous aneurysmal cysts indicating its probable neoplastic nature. Our case had unique features like multiplicity and occurrence after resection of primary aneurysmal bone cyst of the underlying bone.

Karkuzhali, P; Bhattacharyya, Mahuya; Sumitha, P

2007-01-01

157

Inhibition by Thyrocalcitonin of Estrogen-Induced Bone Resorption in the Mouse Pubic Symphysis  

PubMed Central

A specific local erosion of the medial edges of the pubic bones is induced by administration of estradiol cyclopentylpropionate (ECP) to thyroparathyroidectomized mice. This paper presents both histochemical and biochemical evidence that porcine thyrocalcitonin (TCT) inhibits this estrogen-mediated resorption of the pubic symphysis. A marked increase in osteoclasts with a resultant increase in demonstrable sites of acid phosphatase (AcP) activity was observed in serial sections of resorbing pubic symphyses from ECP-treated animals. Bioassay of excised pubic symphyses revealed a concomitant increase in AcP activity. When mice were treated with TCT in combination with ECP, a marked decrease in osteoclasts with a resultant decrease in demonstrable sites of AcP activity and inhibition of the resorption process occurred. The mouse pubic symphysis thus appears to offer a model for exploring cellular mechanisms by which TCT regulates bone metabolism. ImagesFig 5Fig 6Fig 7Fig 8Fig 9Fig 10Fig 11Fig 12Fig 1Fig 2Fig 3Fig 4

Steinetz, B. G.; Matthews, J. R.; Butler, M. C.; Thompson, S. W.

1973-01-01

158

Raman spectroscopy detects deterioration in biomechanical properties of bone in a glucocorticoid-treated mouse model of rheumatoid arthritis  

NASA Astrophysics Data System (ADS)

Although glucocorticoids are frequently prescribed for the symptomatic management of inflammatory disorders such as rheumatoid arthritis, extended glucocorticoid exposure is the leading cause of physician-induced osteoporosis and leaves patients at a high risk of fracture. To study the biochemical effects of glucocorticoid exposure and how they might affect biomechanical properties of the bone, Raman spectra were acquired from ex vivo tibiae of glucocorticoid- and placebo-treated wild-type mice and a transgenic mouse model of rheumatoid arthritis. Statistically significant spectral differences were observed due to both treatment regimen and mouse genotype. These differences are attributed to changes in the overall bone mineral composition, as well as the degree of phosphate mineralization in tibial cortical bone. In addition, partial least squares regression was used to generate a Raman-based prediction of each tibia's biomechanical strength as quantified by a torsion test. The Raman-based predictions were as accurate as those produced by microcomputed tomography derived parameters, and more accurate than the clinically-used parameter of bone mineral density. These results suggest that Raman spectroscopy could be a valuable tool for monitoring bone biochemistry in studies of bone diseases such as osteoporosis, including tests of drugs being developed to combat these diseases.

Maher, Jason R.; Takahata, Masahiko; Awad, Hani A.; Berger, Andrew J.

2011-08-01

159

A close examination of genes within quantitative trait loci of bone mineral density in whole mouse genome.  

PubMed

Bone mineral density (BMD) is one of the strongest determinants of osteoporotic fracture risk. Over the last decade, a large number of quantitative trait loci (QTL) that regulate BMD have been identified using the mouse model. In an attempt to examine the relationship between those QTL and gene distribution in the mouse genome, we searched PubMed with keywords bone and QTL for every publication up to January 2007; we obtained a total of 75 QTL of BMD. We next obtained genes within a QTL for measurements of BMD from the Ensembl database. We then evaluated the potential connection of every gene with bone biology with Online Mendelian Inheritance in Man (OMIM) and PubMed by using eight key words: bone mineral density, BMD, bone strength, bone size, osteoporosis, osteoblast, osteoclast, and fracture. We obtained a total of 15,084 genes for 75 BMD QTL covering 1,211,376,097 base pairs of genomic sequence. Although this very large number of genes exists within QTL regions, only 291 were identified as candidate genes according to our bioinformatics search. Importantly, the association between polymorphism of many candidate genes and BMD has been reported in human studies. Thus, updated genome information and resources should provide new insight for gene identification of QTL. Accordingly, the comprehensive search of candidate genes in the genome for known QTL may provide unexpected benefits for QTL studies. PMID:18652562

Xiong, Qing; Han, Caili; Beamer, Wesley G; Gu, Weikuan

2008-01-01

160

Establishment and characterization of mouse bone marrow-derived mast cell hybridomas  

SciTech Connect

Interleukin (IL)-3-dependent mouse bone marrow-derived mast cells (BMMCs) are an important model for studying the function of mucosal-type mast cells. In the present study, BMMCs were successfully immortalized by cell fusion using a hypoxanthine-aminopterin-thymidine medium-sensitive variant of P815 mouse mastocytoma (P815-6TgR) as a partner cell line. The established mouse mast cell hybridomas (MMCHs) expressed {alpha}, {beta}, and {gamma} subunits of high-affinity immunoglobulin E (IgE) receptor (Fc{epsilon}RI) and possessed cytoplasmic granules devoid of or partially filled with electron-dense material. Four independent MMCH clones continuously proliferated without supplemental exogenous IL-3 and showed a degranulation response on stimulation with IgE+antigen. Furthermore, histamine synthesis and release by degranulation were confirmed in MMCH-D5, a MMCH clone that showed the strongest degranulation response. MMCH-D5 exhibited elevated levels of IL-3, IL-4, IL-13, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor (TNF)-{alpha}, and cyclooxygenase 2, and production of prostaglandin D{sub 2} and leukotriene C{sub 4} in response to IgE-induced stimulation. MMCH clones also expressed Toll-like receptors (TLRs) 1, 2, 4, and 6 and showed elevated levels of TNF-{alpha} expression in response to stimulation with TLR2 and TLR4 ligands. The MMCHs established using this method should be suitable for studies on Fc{epsilon}RI- and TLR-mediated effector functions of mast cells.

Kawahara, Takeshi, E-mail: tkawafb@shinshu-u.ac.jp [Integrated Department of Sciences of Functional Foods, Graduate School of Agriculture, Shinshu University, Nagano (Japan)] [Integrated Department of Sciences of Functional Foods, Graduate School of Agriculture, Shinshu University, Nagano (Japan)

2012-11-01

161

Cranial irradiation induces bone marrow-derived microglia in adult mouse brain tissue.  

PubMed

Postnatal hematopoietic progenitor cells do not contribute to microglial homeostasis in adult mice under normal conditions. However, previous studies using whole-body irradiation and bone marrow (BM) transplantation models have shown that adult BM cells migrate into the brain tissue and differentiate into microglia (BM-derived microglia; BMDM). Here, we investigated whether cranial irradiation alone was sufficient to induce the generation of BMDM in the adult mouse brain. Transgenic mice that express green fluorescent protein (GFP) under the control of a murine stem cell virus (MSCV) promoter (MSCV-GFP mice) were used. MSCV-GFP mice express GFP in BM cells but not in the resident microglia in the brain. Therefore, these mice allowed us to detect BM-derived cells in the brain without BM reconstitution. MSCV-GFP mice, aged 8-12 weeks, received 13.0 Gy irradiation only to the cranium, and BM-derived cells in the brain were quantified at 3 and 8 weeks after irradiation. No BM-derived cells were detected in control non-irradiated MSCV-GFP mouse brains, but numerous GFP-labeled BM-derived cells were present in the brain stem, basal ganglia and cerebral cortex of the irradiated MSCV-GFP mice. These BM-derived cells were positive for Iba1, a marker for microglia, indicating that GFP-positive BM-derived cells were microglial in nature. The population of BMDM was significantly greater at 8 weeks post-irradiation than at 3 weeks post-irradiation in all brain regions examined. Our results clearly show that cranial irradiation alone is sufficient to induce the generation of BMDM in the adult mouse. PMID:24706998

Okonogi, Noriyuki; Nakamura, Kazuhiro; Suzuki, Yoshiyuki; Suto, Nana; Suzue, Kazutomo; Kaminuma, Takuya; Nakano, Takashi; Hirai, Hirokazu

2014-07-01

162

Lithium treatment elongates primary cilia in the mouse brain and in cultured cells  

SciTech Connect

The molecular mechanisms underlying the therapeutic effects of lithium, a first-line antimanic mood stabilizer, have not yet been fully elucidated. Treatment of the algae Chlamydomonas reinhardtii with lithium has been shown to induce elongation of their flagella, which are analogous structures to vertebrate cilia. In the mouse brain, adenylyl cyclase 3 (AC3) and certain neuropeptide receptors colocalize to the primary cilium of neuronal cells, suggesting a chemosensory function for the primary cilium in the nervous system. Here we show that lithium treatment elongates primary cilia in the mouse brain and in cultured cells. Brain sections from mice chronically fed with Li{sub 2}CO{sub 3} were subjected to immunofluorescence study. Primary cilia carrying both AC3 and the receptor for melanin-concentrating hormone (MCH) were elongated in the dorsal striatum and nucleus accumbens of lithium-fed mice, as compared to those of control animals. Moreover, lithium-treated NIH3T3 cells and cultured striatal neurons exhibited elongation of the primary cilia. The present results provide initial evidence that a psychotropic agent can affect ciliary length in the central nervous system, and furthermore suggest that lithium exerts its therapeutic effects via the upregulation of cilia-mediated MCH sensing. These findings thus contribute novel insights into the pathophysiology of bipolar mood disorder and other psychiatric diseases.

Miyoshi, Ko, E-mail: miyoshi@cc.okayama-u.ac.jp [Department of Brain Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikatacho, Okayama 700-8558 (Japan)] [Department of Brain Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikatacho, Okayama 700-8558 (Japan); Kasahara, Kyosuke; Miyazaki, Ikuko; Asanuma, Masato [Department of Brain Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikatacho, Okayama 700-8558 (Japan)] [Department of Brain Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikatacho, Okayama 700-8558 (Japan)

2009-10-30

163

Translocation frequencies and chromosomal proximities for selected mouse chromosomes in primary B lymphocytes.  

PubMed

Chromosome positions within the nucleus of mammalian cells are nonrandom and it is assumed that chromosomal neighborhoods affect the probability of translocations. Four chromosomes can be involved in c-myc-activating chromosomal translocations in mouse plasmacytoma (PCT): the c-myc gene on mouse chromosome 15 can be juxtaposed to either one of the immunoglobulin (Ig) loci on chromosomes 12 (IgH), 16 (Ig?), or 6 (Ig?). In the BALB/c mouse, the translocation between chromosomes 12 and 15, T(12;15), is most common (90%) while the other two possible translocations, T(6;15) and T(16;15), are much less common (<10%). In contrast, in the BALB/cRb6.15 mouse, T(6;15) is found with the same frequency as T(12;15). We, therefore, examined the distance between chromosomes 15 and 12, 6, and 16 in primary mouse B lymphocytes in order to examine the effect of the chromosome proximity on the translocation frequency. We performed three-dimensional fluorescent in situ hybridization (3D-FISH) with chromosome paints. We acquired three-dimensional image stacks with 90 slices per stack and used constrained iterative deconvolution. The nucleus and chromosomes were segmented from this image stack and the interchromosomal distances were measured. Chromosomes 6 and 15 were found in close proximity in BALB/cRb6.15 mice (82%), whereas they did not share this neighborhood relationship in BALB/c mice. No other chromosome combinations showed such a high percentage of close proximities in either mouse strain. Chromosome positions contribute to translocation frequencies in mouse PCTs. The BALB/cRb6.15 mouse data argue for a proximity relationship of chromosomes that engage in illegitimate recombination. These positions are not, however, the only contributing factor as the T(12;15) translocation preference in BALB/c mice could not be supported by significantly elevated proximity of chromosomes 12 and 15 versus 12 and 16 or 12 and 6. Moreover, while there is a significant increase in T(6;15) in BALB/cRb6.15 mice, T(12;15) still occurs in this mouse strain. PMID:21387545

Righolt, Christiaan H; Wiener, Francis; Taylor-Kashton, Cheryl; Harizanova, Jana; Vermolen, Bart J; Garini, Yuval; Young, Ian T; Mai, Sabine

2011-04-01

164

The relief of bone pain in primary biliary cirrhosis with calcium infusions  

PubMed Central

Intravenous calcium infusions produced subjective relief of bone pain in 14 patients with primary biliary cirrhosis. The bone pain had developed despite long-term parenteral vitamin D therapy. The pain returned after two to three months, but a subsequent course of infusions again brought relief. Before treatment satisfactory iliac crest bone biopsies were obtained in 11 of the patients and were normal in seven; two patients had biopsies indicating osteomalacia and two osteoporosis. After treatment a repeat biopsy in one of the patients with osteomalacia showed marked reduction in osteoid. The infusion treatment produced no change in plasma calcium concentration, serum phosphate, or serum alkaline phosphatase. Absorption of oral calcium was also unchanged.

Ajdukiewicz, A. B.; Agnew, J. E.; Byers, P. D.; Wills, M. R.; Sherlock, Sheila

1974-01-01

165

Imaging primary mouse sarcomas after radiation therapy using cathepsin-activatable fluorescent imaging agents  

PubMed Central

Purpose Cathepsin-activated fluorescent probes can detect tumors in mice and in canine patients. We previously showed that these probes can detect microscopic residual sarcoma in the tumor bed of mice during gross total resection. Many patients with soft tissue sarcoma (STS), and other tumors, undergo radiation therapy (RT) prior to surgery. This study assesses the effect of RT on the ability of cathepsin-activated probes to differentiate between normal and cancerous tissue. Methods and Materials A genetically engineered mouse model of STS was used to generate primary hind limb sarcomas that were treated with hypofractionated RT. Mice were injected intravenously with cathepsin-activated fluorescent probes and various tissues, including the tumor, were imaged using a handheld imaging device. Resected tumor and normal muscle samples were harvested to assess cathepsin expression by western blot. Uptake of activated probe was analyzed by flow cytometry and confocal microscopy. Parallel in vitro studies using mouse sarcoma cells were performed. Results RT of primary STS in mice and mouse sarcoma cell lines caused no change in probe activation or cathepsin protease expression. Increasing radiation dose resulted in an upward trend in probe activation. Flow cytometry and immunofluorescence showed that a substantial proportion of probe-labeled cells were CD11b positive tumor associated immune cells. Conclusions In this primary mouse model of STS, RT does not affect the ability of cathepsin-activated probes to differentiate between tumor and normal muscle. Cathepsin-activated probes label tumor cells and tumor associated macrophages. Our results support including patients who have received preoperative RT in clinical studies evaluating cathepsin-activated imaging probes.

Cuneo, Kyle C.; Mito, Jeffrey K.; Javid, Melodi P.; Ferrer, Jorge M.; Kim, Yongbaek; Lee, W. David; Bawendi, Moungi G.; Brigman, Brian E.; Kirsch, David G.

2014-01-01

166

Cloning and initial characterization of mouse meltrin beta and analysis of the expression of four metalloprotease-disintegrins in bone cells.  

PubMed

Here we report the cloning and initial biochemical characterization of the mouse metalloprotease/disintegrin/cysteine-rich (MDC) protein meltrin beta and the analysis of the mRNA expression of four MDC genes (meltrin alpha, meltrin beta, mdc9, and mdc15) in bone cells, including osteoclasts and osteoblasts. Like most other MDC proteins, the predicted meltrin beta protein consists of a signal sequence, prodomain, metalloprotease domain with a predicted catalytic site, disintegrin domain, cysteine-rich region, epidermal growth factor repeat, transmembrane domain, and cytoplasmic domain with putative signaling motifs, such as potential SH3 ligand domains. Northern blot analysis indicates that meltrin beta is widely expressed, with the highest expression in bone, heart, and lung. RNase protection studies revealed expression of all four MDC genes analyzed here in osteoblasts, whereas only mdc9 and mdc15 mRNAs were detectable in osteoclast-like cells generated in vitro. Treatment of primary osteoblasts with 10 nM calcitriol increased meltrin beta expression more than 3-fold, and both meltrin alpha and meltrin beta expression is apparently regulated in a differentiation-associated manner in a mouse osteoblastic cell line, MC3T3E1. Collectively, these results suggest that meltrin alpha and meltrin beta may play a role in osteoblast differentiation and/or function but are not likely to be involved in osteoclast fusion. PMID:9461614

Inoue, D; Reid, M; Lum, L; Krätzschmar, J; Weskamp, G; Myung, Y M; Baron, R; Blobel, C P

1998-02-13

167

The Pathophysiology of Primary Hip Osteoarthritis may Originate from Bone Alterations  

PubMed Central

Objectives: The aim of this study was to investigate whether bone alterations detected by hip magnetic resonance imaging (MRI) were associated with subsequent primary hip OA. Methods: We enrolled 7 patients with hip joint pain from their first visit, at which hip joints were classified as grade 0 or I on the Kellgren-Lawrence grading scale. Plain radiographs and magnetic resonance imaging (MRI) were performed on all cases, and pain was assessed with the Denis pain scale. Average age, height, weight, body mass index, bone mineral density (L1-4), central edge angle, Sharp’s angle, and acetabular hip index were calculated. Results: Within two months of the onset of pain, 4 of the 7 cases showed broad bone signal changes, while 3 cases showed local signal changes in the proximal femur on hip MRI. Three to 6 months after the onset of pain, in all patients whose pain was much improved, plain radiographs showed progression to further-stage OA. Conclusion: Our findings suggest that bone abnormalities in the proximal femur might be involved in the pathogenesis of primary hip OA.

Kamimura, Mikio; Nakamura, Yukio; Ikegami, Shota; Mukaiyama, Keijiro; Uchiyama, Shigeharu; Kato, Hiroyuki

2013-01-01

168

Primary giant cell tumours of the digital bones of the hand.  

PubMed

Primary giant cell tumours involving digital bones of the hand are rare lesions which are generally diagnosed at an advanced stage. Accurate diagnosis requires clinical evaluation, imaging studies and histopathological assessment. Conservative treatment by digit-sparing surgery is associated with high recurrence rates. In a ten year retrospective review, this study identified only four cases. Three cases involved a phalanx and were treated by distal amputation of the involved digit. None recurred. One involved the metacarpal and recurred twice following repeated curettage and bone grafting. No further recurrence has been detected after resection and replacement with a non-vascularised fibular graft and Silastic implant replacement of the metacarpophalangeal joint. Our small series of cases supports a policy of aggressive primary surgery, including amputation or en bloc resection and reconstruction. PMID:17222953

Ropars, M; Kaila, R; Cannon, S R; Briggs, T W R

2007-04-01

169

Vaccination with DKK1-derived Peptides Promotes Bone Formation and Bone Mass in an Aged Mouse Osteoporosis Model.  

PubMed

The investigation of agents for the treatment of osteoporosis has been a long-standing effort. The Wnt pathway plays an important role in bone formation and regeneration, and expression of Wnt pathway inhibitors, Dickkopf-1 (DKK1), appears to be associated with changes in bone mass. Inactivation of DKK1 leads to substantially increased bone mass in genetically manipulated animals. DKK1-derived peptides (DDPs) were added to BMP2-stimulated MC3T3-E1 preosteoblastic cells in vitro to evaluate inhibitory activity of DDPs in MC3T3-E1 cell differentiation. Study was extended in vivo on old female mice to show whether or not inhibition of endogenous DKK1 biological activity using DDPs vaccination approach leads to increase of bone formation, bone density, and improvement of bone microstructure. We reported that synthetic DDPs were able to reduce alkaline phosphatase activity, prevent mineralization and inhibit the differentiation of MC3T3-E1 cells in vitro. Furthermore, vaccination with these DDPs in aged female mice 4 times for a total period of 22 weeks promoted bone mass and bone microstructure. 3D microCT and histomorphometric analysis showed that there were significant increase in bone mineral densities, improvement of bone microstructure and promotion of bone formation in the vaccinated mice, especially in the mice vaccinated with DDP-A and DDP-C. Histological and scanning electron microscopy image analysis also indicated that vaccination increased trabecular bone mass and significantly decreased fragmentation of bone fibers. Taken together, these preclinical results suggest that vaccination with DDPs represents a promising new therapeutic approach for the treatment of bone-related disorders, such as osteoporosis. PMID:24907907

Wu, Qiong; Li, Rui-Shu; Zhao, Yue; Wang, Zhi-Xia; Tang, Yan-Chun; Zhang, Jing; Liu, Jian-Ning; Tan, Xiang-Yang

2014-08-01

170

Primary bone lymphoma of the left radius: a case report and related literature review  

PubMed Central

Primary bone lymphoma (PBL) is a rare but distinct clinicopathological disease. Because it is not common, the optimal treatment strategy has not been established. Here, we present a patient with PBL of the left radius and review the related literature. We focus on the standard treatment for PBL. Many aspects such as rehabilitation, local control and overall survival need to be considered. Studies on this disease should be carried out to clarify the optimal treatment in the future.

2014-01-01

171

Magnesium Deficiency: Effect on Bone and Mineral Metabolism in the Mouse  

Microsoft Academic Search

Insufficient dietary magnesium (Mg) intake has been associated in humans with low bone mass. Mg deficiency in the rat has suggested bone loss is due to increased bone resorption and\\/or inadequate bone formation during remodeling. The purpose of this study was to assess the effect of a low Mg diet on bone and mineral metabolism in the young and mature

R. K. Rude; H. E. Gruber; L. Y. Wei; A. Frausto; B. G. Mills

2003-01-01

172

Establishment of an osteogenic cell line derived from adult mouse bone marrow stroma by use of a recombinant retrovirus.  

PubMed

In order to characterize fibroblastic colony-forming units (CFU-F) from murine bone marrow in relation to osteogenesis, adherent cells of 7-day-old BALB/c mouse bone marrow cultures were infected with a recombinant retrovirus (N2/ delta fosB) containing the bacterial neomycin resistance gene. One of the G418-resistant clones, MN7, was selected for further analysis on the basis of its high expression of the bone-specific alkaline phosphatase. The cells have now been in culture for more than 1 year and maintain a stable phenotype. The osteogenic nature of the immortalized clone MN7 was demonstrated as follows: (1) Mineralization was detected by 85Sr uptake and with the Von Kossa staining method only after in vitro cultivation on a collagen type I matrix. (2) Osteoblastic phenotype markers, including the synthesis of type I collagen, osteonectin, and the bone-specific isoenzyme of alkaline phosphatase were expressed in vitro. (3) MN7 cells responded to bone effectors such as parathyroid hormone and 1,25-dihydroxyvitamin D3. (4) Intraperitoneal injection of MN7 cells into 1-day-old BALB/c mice produced typical osteosarcomas in all animals. We conclude that MN7, derived entirely in vitro from a stromal CFU-F colony, represents a stable murine osteosarcoma cell line expressing the osteoblastic phenotype and provides the first direct evidence needed to establish adult mouse marrow-derived, nonhematopoietic stromal cells as osteoprogenitors. PMID:1571849

Mathieu, E; Schoeters, G; vander Plaetse, F; Merregaert, J

1992-04-01

173

A Systems Biology Study on NF?B Signaling in Primary Mouse Hepatocytes  

PubMed Central

The cytokine tumor necrosis factor-alpha (TNF?) is one of the key factors during the priming phase of liver regeneration as well as in hepatocarcinogenesis. TNF? activates the nuclear factor ?-light-chain-enhancer of activated B cells (NF?B) signaling pathway and contributes to the conversion of quiescent hepatocytes to activated hepatocytes that are able to proliferate in response to growth factor stimulation. Different mathematical models have been previously established for TNF?/NF?B signaling in the context of tumor cells. Combining these mathematical models with time-resolved measurements of expression and phosphorylation of TNF?/NF?B pathway constituents in primary mouse hepatocytes revealed that an additional phosphorylation step of the NF?B isoform p65 has to be considered in the mathematical model in order to sufficiently describe the dynamics of pathway activation in the primary cells. Also, we addressed the role of basal protein turnover by experimentally measuring the degradation rate of pivotal players in the absence of TNF? and including this information in the model. To elucidate the impact of variations in the protein degradation rates on TNF?/NF?B signaling on the overall dynamic behavior we used global sensitivity analysis that accounts for parameter uncertainties and showed that degradation and translation of p65 had a major impact on the amplitude and the integral of p65 phosphorylation. Finally, our mathematical model of TNF?/NF?B signaling was able to predict the time-course of the complex formation of p65 and of the inhibitor of NF?B (I?B) in primary mouse hepatocytes, which was experimentally verified. Hence, we here present a mathematical model for TNF?/NF?B signaling in primary mouse hepatocytes that provides an important basis to quantitatively disentangle the complex interplay of multiple factors in liver regeneration and tumorigenesis.

Pinna, Federico; Sahle, Sven; Beuke, Katharina; Bissinger, Michaela; Tuncay, Selcan; D'Alessandro, Lorenza A.; Gauges, Ralph; Raue, Andreas; Timmer, Jens; Klingmuller, Ursula; Schirmacher, Peter; Kummer, Ursula; Breuhahn, Kai

2012-01-01

174

Reducing the risk of deep wound infection in primary joint arthroplasty with antibiotic bone cement.  

PubMed

Despite significant advances in intraoperative antimicrobial procedures, deep wound infection remains the most serious complication associated with primary, cemented total joint arthroplasty. A systematic review was conducted to evaluate studies of antibiotic bone cement prophylaxis for reducing the risk of deep wound infection. The literature included 22 articles providing estimates of the prophylactic effectiveness of antibiotic cement. In reducing deep wound infection, antibiotic cement was consistently superior to plain cement, similar to systematic antiobiotics, and independent and additive in effect when combined with other prophylactic measures. Randomized controlled trials in particular had important methodological limitations. However, the collective results nearly unanimously favored prophylactic use of antibiotic cement in primary arthoplasty procedures. PMID:16295192

Block, Jon E; Stubbs, Harrison A

2005-11-01

175

[Magnetic resonance in primary bone tumors: a review of 10 years of activities].  

PubMed

All the MR exams of primary bone tumors performed during ten years were reviewed by three different radiologists. In all, 484 exams in 220 patients were considered--namely, 160 exams (33.1%) for staging purposes, 219 (45.2%) during therapy and 105 (21.7%) performed more than 8 months after the last treatment. Its well-known accuracy in the assessment of intra/extraosseous spread confirms the major role of MRI in the staging of primary bone tumors. During treatment, the overall accuracy of this method decreased to 88.8% because of the presence of therapy-induced tissue changes. MRI was 95.2% reliable in the detection of persistent disease or relapse in the exams performed long after therapy. Conventional radiology is still the method of choice in the study of primary bone tumors at presentation as it detects the lesion, differentiates malignant tumors and usually suggests the possible histotype. Nevertheless, MRI seems to be needed to depict actual tumor extent and to find the correct therapeutic approach. In the follow-up, MRI is the best single method to assess the response to therapy and to detect tumor persistence. PMID:8643841

Balzarini, L; Sicilia, A; Ceglia, E; Tesoro Tess, J D; Trecate, G; Musumeci, R

1996-04-01

176

Prolonged Bioluminescence Monitoring in Mouse Ex Vivo Bone Culture Revealed Persistent Circadian Rhythms in Articular Cartilages and Growth Plates  

PubMed Central

The bone is a metabolically active organ which undergoes repeated remodeling cycles of bone resorption and formation. In this study, we revealed a robust and extremely long-lasting circadian rhythm in ex vivo culture maintained for over six months from the femoral bone of a PERIOD2Luciferase mouse. Furthermore, we also identified robust circadian clocks in flat bones. High- or low-magnification real-time bioluminescence microscopic imaging revealed that the robust circadian rhythms emanated from the articular cartilage and the epiphyseal cartilage within the growth plate of juvenile animals. Stimulation by forskolin or dexamethasone treatment caused type 0 phase resetting, indicating canonical entraining properties of the bone clock. Together, our findings from long-term ex vivo culture revealed that “tissue-autonomous” circadian rhythm in the articular cartilage and the growth plate of femoral bone functions for several months even in an organ culture condition, and provided a useful in vitro assay system investigating the role of the biological clock in bone formation or development.

Fujiwara, Hiroyoshi; Umemura, Yasuhiro; Tsuchiya, Yoshiki; Shirai, Toshiharu; Oda, Ryo; Inokawa, Hitoshi; Kubo, Toshikazu; Yagita, Kazuhiro

2013-01-01

177

Primary Neoplasms of Bones in Mice: Retrospective Study and Review of Literature  

PubMed Central

To compare and summarize the mechanisms, frequencies of occurrence, and classification schemes of spontaneous, experimental, and genetically engineered, mouse skeletal neoplasms, the literature was reviewed and archived case material at The Jackson Laboratory examined. The frequency of occurrence of spontaneous bone neoplasms was less than 1% for most strains, with the exceptions of osteomas in CF-1 (5.5% and 10% in two studies) and OF-1 outbred strains (35%), and osteosarcomas in NOD/ShiLtJ (11.5%) and NOD derived (7.1%) mice. The frequency was 100% for osteochondromas induced by conditional inactivation of exostoses (multiple) 1 (Ext1) in chondrocytes, osteosarcomas induced by tibial intramedullary inoculation of Moloney’s murine sarcoma virus, and osteosarcomas induced by conditional inactivation of Trp53-with or without inactivation of Rb1-in osteoblast precursors. Spontaneous osteogenic neoplasms were more frequent than spontaneous cartilaginous and vascular types. Malignant neoplasms were more frequent than benign ones. The age of occurrence for spontaneous neoplasms ranged from 37 to 720 (Mean 316.35) days for benign, and 35 to 990 (Mean 299.28) days for malignant neoplasms. In genetically engineered mice, the average age of occurrence ranged from 28 to 70 days for benign, and from 35 to 690 days for malignant neoplasms. Histologically, non-osteogenic neoplasms were similar across strains and mutant stocks; osteogenic neoplasms exhibited greater diversity. This comparison and summarization of mouse bone neoplasms provides valuable information for the selection of strains to create, compare, and validate models of bone neoplasms.

Kavirayani, A. M.; Sundberg, J. P.; Foreman, O.

2011-01-01

178

Cyclosporine Increases Human Immunodeficiency Virus Type 1 Vector Transduction of Primary Mouse Cells  

PubMed Central

Murine primary cells are poorly permissive to human immunodeficiency virus type 1 (HIV-1) vector infection. Retroviral infectivity is influenced by dominant inhibitors such as TRIM5?. Sensitivity to TRIM5? is altered by interactions between cyclophilin A and the HIV-1 capsid. Here we demonstrate that competitive inhibitors of cyclophilins, cyclosporine or the related Debio-025, stimulate HIV-1 vector transduction of primary murine cells, including bone marrow and macrophages, up to 20-fold. Unexpectedly, the infectivity of an HIV-1 mutant or a simian lentivirus that does not recruit cyclophilin A is also stimulated by these drugs. We propose that cyclosporine and related compounds will be useful tools for experimental infection of murine primary cells. It is possible that HIV-1 infection of murine cells is inhibited by dominant factors related to immunophilins.

Noser, Josh A.; Towers, Greg J.; Sakuma, Ryuta; Dumont, Jean-Maurice; Collins, Mary K. L.; Ikeda, Yasuhiro

2006-01-01

179

[Effect of alloreactive natural killer cells on immune reconstitution in mouse haploidentical bone marrow transplantation].  

PubMed

The study was purposed to investigate the effect of alloreactive natural killer (alloNK) cells on immune reconstitution in murine haploidentical bone marrow transplantation (BMT). The murine model of haploidentical BMT was established by using (C57BL/6×BALB/c)BCF(1)(H-2(d/b)) mouse as the donor, and BALB/c (H-2(d)) mouse as the recipient. Recipient mice were divided into BMT group, non-allo-reactive NK (non-alloNK) cell group and alloNK cell group according to different transfusion. The effect of adding alloNK cells to transfusion was assessed by thymus pathology, the proportion of spleen NK cells, the spleen cell proliferation, the IFN-? and IL-4 concentrations product at 24 and 48 h of recipient spleen cell culture supernatant at 2 months after BMT. The results showed that there were no obvious difference in thymus tissue among 3 groups under the optical microscope. The proportion of recipient spleen NK cells in non-alloNK group was significantly lower than that in BMT group (P < 0.05). There was no significant difference in proliferation of the recipient spleen cells among 3 groups at 2 months after BMT. The IFN-? concentration product at 24 and 48 h of recipient spleen cell culture supernatant in alloNK group was significantly lower than that in other 2 groups at 2 months after BMT (P < 0.05). The IL-4 concentration in each group was not significantly different (P > 0.05). It is concluded that alloNK cells do not damage the thymus structure and may induce Th2 immune response in murine haploidentical BMT. PMID:23114142

Wang, Hua; Wang, Hui; Liu, Ying-Hui; Feng, Si-Zhou; Han, Ming-Zhe

2012-10-01

180

Isolation and characterization of mouse bone marrow-derived Lin?/VEGF-R2? progenitor cells.  

PubMed

Circulating endothelial progenitor cells (EPCs) in the peripheral blood (PB) have physiological roles in the maintenance of the existing vascular beds and rescue of vascular injury. In this study, we have evaluated the properties of Lin?/VEGF-R2? progenitor cells isolated from the mouse bone marrow (BM) and further studied their distribution and integration in an animal model of laser-induced retinal vascular injury. Lin?/VEGF-R2? cells were enriched from C57BL/6 mice BM using magnetic cell sorting with hematopoietic lineage (Lin) depletion followed by VEGF-R2 positive selection. Lin?/VEGF-R2? BM cells were characterized using flow cytometry and immunocytochemistry and further tested for colony formation during culture and tube formation on Matrigel®. Lin?/VEGF-R2? BM cells possessed typical EPC properties such as forming cobble-stone shaped colonies after 3 to 4 weeks of culture, CD34? expression, take up of Dil-acLDL and binding to Ulex europaeus agglutinin. However, they did not form tube-like structures on Matrigel®. The progenitor cells retained their phenotype over extended period of culture. After intravitreal transplantation in eyes subjected to the laser-induced retinal vascular injury, some Lin?/VEGF-R2? cells were able to integrate into the damaged retinal vasculature but the level of cell integration seemed less efficient when compared with previous reports in which EPCs from the human PB were employed. Our results indicate that Lin?/VEGF-R2? cells isolated from the mouse BM share some similarities to EPCs from the human PB but most of them are at a very early stage of maturation and remain quiescent during culture and after intravitreal transplantation. PMID:23771478

Barthelmes, Daniel; Irhimeh, Mohammad R; Gillies, Mark C; Zhu, Ling; Shen, Weiyong

2013-11-01

181

RANKL\\/RANK\\/osteoprotegerin system as novel therapeutic target in the treatment of primary bone tumors and osteolytic metastases  

Microsoft Academic Search

Primary bone tumors and cancers that metastasize to bone require osteoclastic activity to release tumor-supportive growth factors from bone tissue. A number of systemic and locally acting factors are known to influence osteoclast formation, fusion, activation, and survival. Recently, two critical extracel- lular regulators of osteoclast differentiation and activation have been identified: receptor activator of nuclear factor (NF-kappaB) ligand (RANKL)

Zlatibor Anðelkoviæ; Vuka Katiæ; Aleksandar Petroviæ

2004-01-01

182

Metal block augmentation for bone defects of the medial tibia during primary total knee arthroplasty  

PubMed Central

Background Stable and well-aligned placement of tibial components during primary total knee arthroplasty is challenging in patients with bone defects. Although rectangular block-shaped augmentations are widely used to reduce the shearing force between the tibial tray and bone compared with wedge-shaped augmentations, the clinical result remains unclear. This study aimed to evaluate the outcome of primary total knee arthroplasty with metal block augmentation. Methods We retrospectively reviewed the 3- to 6-year follow-up results of 33 knees that underwent total knee arthroplasty with metal block augmentation (metal-augmented group) for bone defects of the medial tibia and 132 varus knees without bone defects as the control group. All surgeries were performed using posterior-stabilized cemented prostheses in both groups. Cemented stems were routinely augmented when the metal block was used. Results There were no differences in implant survival rates (100% in metal-augmented and 99.2% in control) or knee function scores (82 points in metal-augmented and 84 points in control) between the two groups at the final follow-up examination (P = 0.60 and P = 0.09, respectively). No subsidence or loosening of the tibial tray was observed. Of 33 metal-augmented total knee arthroplasties, a nonprogressive radiolucent line beneath the metal was detected in 10 knees (30.3%), and rounding of the medial edge of the tibia was observed in 17 knees (51.5%). Conclusions The clinical results of total knee arthroplasty with metal augmentation were not inferior to those in patients without bone defects. However, radiolucent lines were observed in 30.3%.

2013-01-01

183

Mouse genome-wide association and systems genetics identify Asxl2 as a regulator of bone mineral density and osteoclastogenesis.  

PubMed

Significant advances have been made in the discovery of genes affecting bone mineral density (BMD); however, our understanding of its genetic basis remains incomplete. In the current study, genome-wide association (GWA) and co-expression network analysis were used in the recently described Hybrid Mouse Diversity Panel (HMDP) to identify and functionally characterize novel BMD genes. In the HMDP, a GWA of total body, spinal, and femoral BMD revealed four significant associations (-log10P>5.39) affecting at least one BMD trait on chromosomes (Chrs.) 7, 11, 12, and 17. The associations implicated a total of 163 genes with each association harboring between 14 and 112 genes. This list was reduced to 26 functional candidates by identifying those genes that were regulated by local eQTL in bone or harbored potentially functional non-synonymous (NS) SNPs. This analysis revealed that the most significant BMD SNP on Chr. 12 was a NS SNP in the additional sex combs like-2 (Asxl2) gene that was predicted to be functional. The involvement of Asxl2 in the regulation of bone mass was confirmed by the observation that Asxl2 knockout mice had reduced BMD. To begin to unravel the mechanism through which Asxl2 influenced BMD, a gene co-expression network was created using cortical bone gene expression microarray data from the HMDP strains. Asxl2 was identified as a member of a co-expression module enriched for genes involved in the differentiation of myeloid cells. In bone, osteoclasts are bone-resorbing cells of myeloid origin, suggesting that Asxl2 may play a role in osteoclast differentiation. In agreement, the knockdown of Asxl2 in bone marrow macrophages impaired their ability to form osteoclasts. This study identifies a new regulator of BMD and osteoclastogenesis and highlights the power of GWA and systems genetics in the mouse for dissecting complex genetic traits. PMID:21490954

Farber, Charles R; Bennett, Brian J; Orozco, Luz; Zou, Wei; Lira, Ana; Kostem, Emrah; Kang, Hyun Min; Furlotte, Nicholas; Berberyan, Ani; Ghazalpour, Anatole; Suwanwela, Jaijam; Drake, Thomas A; Eskin, Eleazar; Wang, Q Tian; Teitelbaum, Steven L; Lusis, Aldons J

2011-04-01

184

Mouse Genome-Wide Association and Systems Genetics Identify Asxl2 As a Regulator of Bone Mineral Density and Osteoclastogenesis  

PubMed Central

Significant advances have been made in the discovery of genes affecting bone mineral density (BMD); however, our understanding of its genetic basis remains incomplete. In the current study, genome-wide association (GWA) and co-expression network analysis were used in the recently described Hybrid Mouse Diversity Panel (HMDP) to identify and functionally characterize novel BMD genes. In the HMDP, a GWA of total body, spinal, and femoral BMD revealed four significant associations (?log10P>5.39) affecting at least one BMD trait on chromosomes (Chrs.) 7, 11, 12, and 17. The associations implicated a total of 163 genes with each association harboring between 14 and 112 genes. This list was reduced to 26 functional candidates by identifying those genes that were regulated by local eQTL in bone or harbored potentially functional non-synonymous (NS) SNPs. This analysis revealed that the most significant BMD SNP on Chr. 12 was a NS SNP in the additional sex combs like-2 (Asxl2) gene that was predicted to be functional. The involvement of Asxl2 in the regulation of bone mass was confirmed by the observation that Asxl2 knockout mice had reduced BMD. To begin to unravel the mechanism through which Asxl2 influenced BMD, a gene co-expression network was created using cortical bone gene expression microarray data from the HMDP strains. Asxl2 was identified as a member of a co-expression module enriched for genes involved in the differentiation of myeloid cells. In bone, osteoclasts are bone-resorbing cells of myeloid origin, suggesting that Asxl2 may play a role in osteoclast differentiation. In agreement, the knockdown of Asxl2 in bone marrow macrophages impaired their ability to form osteoclasts. This study identifies a new regulator of BMD and osteoclastogenesis and highlights the power of GWA and systems genetics in the mouse for dissecting complex genetic traits.

Farber, Charles R.; Bennett, Brian J.; Orozco, Luz; Zou, Wei; Lira, Ana; Kostem, Emrah; Kang, Hyun Min; Furlotte, Nicholas; Berberyan, Ani; Ghazalpour, Anatole; Suwanwela, Jaijam; Drake, Thomas A.; Eskin, Eleazar; Wang, Q. Tian; Teitelbaum, Steven L.; Lusis, Aldons J.

2011-01-01

185

Long-term primary culture of mouse mammary tumor cells: production of virus.  

PubMed

Long-term primary cultures of mouse mammary tumor cells proved an excellent source of mouse mammary tumor virus (MMTV). Virus purified from these primary cultures had the same morphologic biochemical, immunologic, and biologic characteristics as MMTV. Quantitation of MMTV-protein equivalents released into the medium was measured by the radioimmunoassay for MMTV. Peak production levels were 20-40 mug MMTV protien equivalents/75-cm-2 flask/24 hours. These cultures produced MMTV for as long as 90 days. MMTV cultivation depended on the initial cell-plating density and hormones. Maximal MMTV release was obtained at a plating density of 1 times 10-6 cells/cm-2 in the presence of insulin and hydrocortisone. Insulin alone gave basal levels of MMTV, and hydrocortisone alone increased MMTV release only three-fold, but insulin and hydrocortisone together effected an eightfold increase in MMTV release. This suggested that hydrocortisone had a primary effect on MMTV release and insulin acted synergistically with hydrocortisone to maximize MMTV release. PMID:165313

Young, L J; Cardiff, R D; Ashley, R L

1975-05-01

186

An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow  

Microsoft Academic Search

As dendritic cells (DC) are rare populations in all organs, their generation from hematopoietic precursors in large quantities has proven critical to study their biology. From murine bone marrow about 5×106 cells at 70% purity are obtained per mouse after 8 days of culture with GM-CSF. We have improved this standard method and routinely achieve a 50-fold higher yield, i.e.,

Manfred B Lutz; Nicole Kukutsch; Alexandra L. J Ogilvie; Susanne Rößner; Franz Koch; Nikolaus Romani; Gerold Schuler

1999-01-01

187

Tissue Inhibitor of Matrix Metalloproteinase-1 Suppresses Apoptosis of Mouse Bone Marrow Stromal Cell Line MBA-1  

Microsoft Academic Search

We investigated the action of tissue inhibitor of metalloproteinase-1 (TIMP-1) on apoptosis and differentiation of mouse bone\\u000a marrow stromal cell line MBA-1. TIMP-1 did not affect alkaline phosphatase (ALP) activity, suggesting that it is not involved\\u000a in osteoblastic differentiation in MBA-1 cells. However, TIMP-1 inhibited MBA-1 apoptosis induced by serum deprivation in\\u000a a dose-dependent manner. Our study also showed increased

L.-J. Guo; X.-H. Luo; H. Xie; H.-D. Zhou; L.-Q. Yuan; M. Wang; E.-Y. Liao

2006-01-01

188

The 5T2 mouse multiple myeloma model: characterization of 5T2 cells within the bone marrow  

Microsoft Academic Search

The transplantable C57BL\\/KaLwRij mouse 5T2 multiple myeloma (MM) is a new animal model for studies on MM in man. Histological examination of the 5T2 MM cells revealed their morphological heterogeneity. In this study we investigated whether this heterogeneity reflects subpopulations of 5T2 MM cells with different biological properties. 5T2 MM bone marrow cells were separated according to their sedimentation velocity

JW Croese; CM Vas Nunes; J Radl; MHM van den Enden-Vieveen; RJ Brondijk; WJ Boersma

1987-01-01

189

Effects of Different Doses of Bone Morphogenetic Protein 4 on Viability and Proliferation Rates of Mouse Embryonic Stem Cells  

Microsoft Academic Search

Received: 20\\/Jul\\/2008, Accepted: 2\\/Nov\\/2008 Objective: In this study, we examined the effect of different doses of bone morpho- genetic protein 4 (BMP4) on CCE mouse embryonic stem cells (ESCs) viability and proliferation rates in order to improve the outcome of induction processes and make a system with highest viability and proliferation rates for further studies on BMP4 roles in multiple

Zohreh Makoolati; Mansoureh Movahedin; Mehdi Forouzandeh-Moghadam

2009-01-01

190

Imaging Primary Mouse Sarcomas After Radiation Therapy Using Cathepsin-Activatable Fluorescent Imaging Agents  

SciTech Connect

Purpose: Cathepsin-activated fluorescent probes can detect tumors in mice and in canine patients. We previously showed that these probes can detect microscopic residual sarcoma in the tumor bed of mice during gross total resection. Many patients with soft tissue sarcoma (STS) and other tumors undergo radiation therapy (RT) before surgery. This study assesses the effect of RT on the ability of cathepsin-activated probes to differentiate between normal and cancerous tissue. Methods and Materials: A genetically engineered mouse model of STS was used to generate primary hind limb sarcomas that were treated with hypofractionated RT. Mice were injected intravenously with cathepsin-activated fluorescent probes, and various tissues, including the tumor, were imaged using a hand-held imaging device. Resected tumor and normal muscle samples were harvested to assess cathepsin expression by Western blot. Uptake of activated probe was analyzed by flow cytometry and confocal microscopy. Parallel in vitro studies using mouse sarcoma cells were performed. Results: RT of primary STS in mice and mouse sarcoma cell lines caused no change in probe activation or cathepsin protease expression. Increasing radiation dose resulted in an upward trend in probe activation. Flow cytometry and immunofluorescence showed that a substantial proportion of probe-labeled cells were CD11b-positive tumor-associated immune cells. Conclusions: In this primary murine model of STS, RT did not affect the ability of cathepsin-activated probes to differentiate between tumor and normal muscle. Cathepsin-activated probes labeled tumor cells and tumor-associated macrophages. Our results suggest that it would be feasible to include patients who have received preoperative RT in clinical studies evaluating cathepsin-activated imaging probes.

Cuneo, Kyle C. [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States)] [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Mito, Jeffrey K.; Javid, Melodi P. [Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States)] [Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States); Ferrer, Jorge M. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States)] [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Kim, Yongbaek [Department of Clinical Pathology, College of Veterinary Medicine, Seoul National University, Seoul (Korea, Republic of)] [Department of Clinical Pathology, College of Veterinary Medicine, Seoul National University, Seoul (Korea, Republic of); Lee, W. David [The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States)] [The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Bawendi, Moungi G. [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States)] [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts (United States); Brigman, Brian E. [Department of Orthopedic Surgery, Duke University School of Medicine, Durham, North Carolina (United States)] [Department of Orthopedic Surgery, Duke University School of Medicine, Durham, North Carolina (United States); Kirsch, David G., E-mail: david.kirsch@duke.edu [Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina (United States); Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina (United States)

2013-05-01

191

Three Na+/Ca2+ exchanger (NCX) variants are expressed in mouse osteoclasts and mediate calcium transport during bone resorption.  

PubMed

The plasma membrane Na(+)/Ca(2+) exchanger (NCX) is a bidirectional transporter that mediates the exchange of Na(+) for Ca(2+) depending on the electrochemical gradients. Mammalian NCXs form a multigene family comprising NCX1, NCX2, and NCX3 isoforms. Although it has been known that NCX1 in rat osteoclasts is coupled with the Na(+)/ H(+) exchanger for regulation of intracellular Ca(2+) concentration ([Ca(2+)](i)), it is unclear what kind of NCX1 variants are expressed and whether the other two NCX isoforms are also present in mouse osteoclasts. To clarify the role of NCXs during bone resorption, we investigated the expression of NCXs, the ion transport via NCXs, and the effects of NCX inhibitors on bone-resorbing activity in mouse osteoclasts. Using RT-PCR, immunocytochemical, and Western blot methods, we detected three splice variants of NCX1 and NCX3, namely NCX1.3, NCX1.41, and NCX3.2. Of these, NCX1.41 is a newly identified splice variant. Low extracellular sodium ([Na(+)](o)) solution increases the intracellular Ca(2+) concentration via NCX transporter in fura-2-loaded osteoclasts. The [Na(+)](o)-free solution-induced [Ca(2+)](i) increase was suppressed by benzyloxyphenyl NCX inhibitors. Bidirectional NCX currents in mouse osteoclasts were recorded using the patch clamp method and could be suppressed with NCX inhibitors. NCX inhibitors also decreased the resorption pit area surrounding osteoclasts in a dose-dependent manner. Furthermore, small interference RNAs targeted against NCX1.3, NCX1.41, and NCX3.2 expressed in mouse osteoclasts suppressed osteoclastic pit formation. These results show that three NCX variants are expressed in mouse osteoclasts and play an important role for Ca(2+) transport and regulation during osteoclastic bone resorption. PMID:17317768

Li, Jing-Ping; Kajiya, Hiroshi; Okamoto, Fujio; Nakao, Akihiro; Iwamoto, Takahiro; Okabe, Koji

2007-05-01

192

Bone Mass Is Preserved and Cancellous Architecture Altered Due to Cyclic Loading of the Mouse Tibia After Orchidectomy*  

PubMed Central

Introduction The study of adaptation to mechanical loading under osteopenic conditions is relevant to the development of osteoporotic fracture prevention strategies. We previously showed that loading increased cancellous bone volume fraction and trabecular thickness in normal male mice. In this study, we tested the hypothesis that cyclic mechanical loading of the mouse tibia inhibits orchidectomy (ORX)-associated cancellous bone loss. Materials and Methods Ten-week-old male C57BL/6 mice had in vivo cyclic axial compressive loads applied to one tibia every day, 5 d/wk, for 6 wk after ORX or sham operation. Adaptation of proximal cancellous and diaphyseal cortical bone was characterized by ?CT and dynamic histomorphometry. Comparisons were made between loaded and nonloaded contralateral limbs and between the limbs of ORX (n = 10), sham (n = 11), and basal (n = 12) groups and tested by two-factor ANOVA with interaction. Results Cyclic loading inhibited bone loss after ORX, maintaining absolute bone mass at age-matched sham levels. Relative to sham, ORX resulted in significant loss of cancellous bone volume fraction (?78%) and trabecular number (?35%), increased trabecular separation (67%), no change in trabecular thickness, and smaller loss of diaphyseal cortical properties, consistent with other studies. Proximal cancellous bone volume fraction was greater with loading (ORX: 290%, sham: 68%) than in contralateral nonloaded tibias. Furthermore, trabeculae thickened with loading (ORX: 108%, sham: 48%). Dynamic cancellous bone histomorphometry indicated that loading was associated with greater mineral apposition rates (ORX: 32%, sham: 12%) and smaller percent mineralizing surfaces (ORX: ?47%, sham: ?39%) in the final week. Loading resulted in greater BMC (ORX: 21%, sham: 15%) and maximum moment of inertia (ORX: 39%, sham: 24%) at the cortical midshaft. Conclusions This study shows that cancellous bone mass loss can be prevented by mechanical loading after hormonal compromise and supports further exploration of nonpharmacologic measures to prevent rapid-onset osteopenia and associated fractures.

Fritton, J Christopher; Myers, Elizabeth R; Wright, Timothy M; van der Meulen, Marjolein CH

2008-01-01

193

Finite element analysis of dental implant neck effects on primary stability and osseointegration in a type IV bone mandible.  

PubMed

The purpose of this study is to investigate the effect of implant neck design and cortical bone thickness by means of 3-D linearly elastic finite element analysis and to analyze primary and secondary stability of clinical evidence based on micromotion and principal stress. Four commercial dental implants, comparable in size, for a type IV bone and mandibular segments were created. Various parameters were considered, including the osseointegration condition (non- and full bonded), force direction (vertical and horizontal) and cortical bone thickness (0.3, 0.5 and 1mm). The force was considered a static load applied at the top of the platform. The magnitudes of the vertical and horizontal loading direction were 500 N and 250 N. Micromotion and principal stresses were employed to evaluate the failure of osseointegration and bone overloading, respectively. The results show that Maximum stress of the peri-implant bone decreased as cortical bone thickness increased. The stress concentration regions were located at the implant neck between the cortical bone and cancellous bone. The micromotion level in full osseointegration is less than that in non-osseointegration and it also decreases as a increasing of cortical bone thickness. Consequently, cortical bone thickness is a key factor for primary stability. PMID:24212038

Huang, You-Min; Chou, I-Chiang; Jiang, Cho-Pei; Wu, Yi-Syun; Lee, Shyh-Yuan

2014-01-01

194

Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments  

PubMed Central

Neutrophils are critical effector cells of the innate immune system. They are rapidly recruited at sites of acute inflammation and exert protective or pathogenic effects depending on the inflammatory milieu. Nonetheless, despite the indispensable role of neutrophils in immunity, detailed understanding of the molecular factors that mediate neutrophils' effector and immunopathogenic effects in different infectious diseases and inflammatory conditions is still lacking, partly because of their short half life, the difficulties with handling of these cells and the lack of reliable experimental protocols for obtaining sufficient numbers of neutrophils for downstream functional studies and adoptive transfer experiments. Therefore, simple, fast, economical and reliable methods are highly desirable for harvesting sufficient numbers of mouse neutrophils for assessing functions such as phagocytosis, killing, cytokine production, degranulation and trafficking. To that end, we present a reproducible density gradient centrifugation-based protocol, which can be adapted in any laboratory to isolate large numbers of neutrophils from the bone marrow of mice with high purity and viability. Moreover, we present a simple protocol that uses CellTracker dyes to label the isolated neutrophils, which can then be adoptively transferred into recipient mice and tracked in several tissues for at least 4 hr post-transfer using flow cytometry. Using this approach, differential labeling of neutrophils from wild-type and gene-deficient mice with different CellTracker dyes can be successfully employed to perform competitive repopulation studies for evaluating the direct role of specific genes in trafficking of neutrophils from the blood into target tissues in vivo .

Swamydas, Muthulekha; Lionakis, Michail S.

2013-01-01

195

Primary malignant giant cell tumor of bone: "dedifferentiated" giant cell tumor.  

PubMed

Well documented examples of primary malignant giant cell tumor of bone (giant cell tumor and concurrent sarcoma arising de novo) are exceedingly rare in the literature. We report a case arising in the left ischium of a 44-yr-old man. He had no previous history of radiation therapy or multiple resections. Histologically, the tumor was a typical giant cell tumor of bone juxtaposed to a malignant fibrous histiocytoma (MFH). The juxtaposition of a high grade sarcoma (MFH) and a locally aggressive nonmalignant neoplasm such as giant cell tumor is analogous to several other tumors of bone and soft tissue in which a low grade malignant or locally aggressive tumor can be associated with MFH or fibrosarcoma de novo, namely chondrosarcoma, chordoma, liposarcoma, and well differentiated intraosseous and parosteal osteosarcoma. The presence of a high grade malignant component in each of the aforementioned neoplasms generally portends a more ominous prognosis, although this is not invariably true. Recognition of the phenomenon of "dedifferentiation" (or tumor progression) in some bone tumors and sarcomas is important to ensure appropriate treatment. Distinction from secondary malignant giant cell tumors which are usually radiation induced is also important, since the latter have a much worse prognosis than those with dedifferentiation occurring de novo. PMID:2554283

Meis, J M; Dorfman, H D; Nathanson, S D; Haggar, A M; Wu, K K

1989-09-01

196

Bone marrow fibrosis in patients with primary myelodysplastic syndromes has prognostic value using current therapies and new risk stratification systems.  

PubMed

Bone marrow fibrosis has recently been recognized as an adverse histological feature in patients with primary myelodysplastic syndromes. In this study, we assessed the prognostic impact of bone marrow fibrosis in patients with primary myelodysplastic syndromes under the recently revised new risk stratification systems: the New Comprehensive Cytogenetic Scoring System and the Revised International Prognostic Scoring System. From 2002 to 2012, a total of 79 (13%) patients with primary myelodysplastic syndromes and moderate/severe bone marrow fibrosis were identified; and these patients were compared with a control group of 166 patients with myelodysplastic syndromes but no significant fibrosis. Bone marrow fibrosis predicted an inferior overall survival and leukemia event-free survival for patients who received no hematopoietic stem cell transplant in univariate and multivariate analysis. Eleven patients with bone marrow fibrosis and 32 control group patients underwent hematopoietic stem cell transplant; and bone marrow fibrosis was an independent risk for an inferior overall survival but not leukemia-free survival. In addition, 17 (4%) patients developed bone marrow fibrosis during the course of myelodysplastic syndromes, which was accompanied by clinical and cytogenetic evidence of disease progression. JAK2 V617F mutations were detected in 6 of the 28 patients with bone marrow fibrosis presenting at the time of diagnosis and 2 of the 7 patients with bone marrow fibrosis developing in the course of disease, significantly higher than the control group patients. We conclude that bone marrow fibrosis is an adverse risk feature in primary myelodysplastic syndromes in the current therapeutic era, and this risk feature is not captured by newly revised risk stratification systems. Inclusion of bone marrow fibrosis in patient assessment may further aid in risk-adapted therapeutic decisions. PMID:24186132

Fu, Bin; Jaso, Jesse M; Sargent, Rachel L; Goswami, Maitrayee; Verstovsek, Srdan; Medeiros, L Jeffrey; Wang, Sa A

2014-05-01

197

Evaluation and validation of multiple cell lines and primary mouse macrophages to predict phospholipidosis potential.  

PubMed

Phospholipidosis (PLD) in preclinical species can lead to regulatory delays thereby creating incentives to screen for PLD during drug discovery. The objective of this work was to compare, optimize, and validate in vitro PLD assays in primary mouse macrophages and hepatocyte- (HepG2, HuH7) or macrophage-derived cells lines (I.13.35, RAW264.7) and to evaluate whether primary cells were better at predicting PLD. Assay precision, determined by a measure of signal to noise window (Z'), within assay variability, and day-to-day variability, using amiodarone, was generally acceptable for all cell types; however, precision limits for HepG2 and HuH7 were slightly below assay acceptance criteria. Up to 66 known PLD inducers and non-inducers were subsequently tested to validate the assays. The concordance for predicting PLD in primary macrophages, I-13.35, RAW264.7, HuH7, and HepG2 cells was 91%, 74%, 73%, 62%, and 62% respectively using a decision limit of EC50?125 ?M as a positive finding. Increasing the number of negative controls tested in RAW264.7 cells and changing the decision limit to ?4-fold increase in PLD, improved the specificity and overall concordance to 88%. RAW264.7 cells were selected as the primary screen for predicting PLD, and together with the primary macrophages, were integrated into an overall testing paradigm proposed for use in PLD risk identification. PMID:21767630

LeCureux, Lloyd; Cheng, Charles S; Herbst, John; Reilly, Timothy P; Lehman-McKeeman, Lois; Otieno, Monicah

2011-12-01

198

Minor histocompatibility antigens on transfused leukoreduced units of red blood cells induce bone marrow transplant rejection in a mouse model  

PubMed Central

When successful, human leukocyte antigen (HLA)–matched bone marrow transplantation with reduced-intensity conditioning is a cure for several nonmalignant hematologic disorders that require chronic transfusion, such as sickle cell disease and aplastic anemia. However, there are unusually high bone marrow transplant (BMT) rejection rates in these patients. Rejection correlates with the number of transfusions before bone marrow transplantation, and it has been hypothesized that preimmunization to antigens on transfused blood may prime BMT rejection. Using a novel mouse model of red blood cell (RBC) transfusion and major histocompatibility complex–matched bone marrow transplantation, we report that transfusion of RBC products induced BMT rejection across minor histocompatibility antigen (mHA) barriers. It has been proposed that contaminating leukocytes are responsible for transfusion-induced BMT rejection; however, filter leukoreduction did not prevent rejection in the current studies. Moreover, we generated a novel transgenic mouse with RBC-specific expression of a model mHA and demonstrated that transfusion of RBCs induced a CD8+ T-cell response. Together, these data suggest that mHAs on RBCs themselves are capable of inducing BMT rejection. Cellular immunization to mHAs is neither monitored nor managed by current transfusion medicine practice; however, the current data suggest that mHAs on RBCs may represent an unappreciated and significant consequence of RBC transfusion.

Desmarets, Maxime; Cadwell, Chantel M.; Peterson, Kenneth R.; Neades, Renee

2009-01-01

199

[The long-term results of primary total knee arthroplasty using bone grafts].  

PubMed

In the presented paper the authors evaluated the results of treatment in patients following primary total knee arthroplasty using bone grafts due to sever valgus or varus deformity. The clinical material from the years 2000-2004 consists of 18 patients aged from 64 to 77; namely, 14 women (77.8%) and 4 men (22.2%). The longest time of follow-up was 3 years and the shortestwas 17 months. The authors evaluated achieved results in the period between 17th and 24th month since total knee arthroplasty in 18 patients on the basis of the time criteria referring to the time of healing the bone implants and on the basis of the modificated HSS knee rating scale. We achieved 15 well, 2 sufficient and 1 insufficient based on time criteria and 9 very well, 5 well, 3 sufficient and 1 insufficient results based on modificated HSS knee rating scale. PMID:17639917

Nowak, Sebastian; Golec, Edward; Widawski, Antoni; Golec, Joanna; Wa?ach, Angelika

2007-01-01

200

Two types of bone resorption lacunae in the mouse parietal bones as revealed by scanning electron microscopy and histochemistry.  

PubMed

To understand the bone resorption process on the basis of the morphology of bone resorption lacunae, the inner surface of parietal bones in juvenile mice was exposed with a treatment of ultrasonic waves or NaOCl treatment and examined by scanning electron microscopy (SEM). The bone resorption lacunae were divided into two types (I and II) according to differences in morphological features of their walls; the wall of type I lacunae was covered with loose collagen fibrils, while that of type II lacunae was smooth with almost no fibrillar structures. Collagen fibrils in type I lacunae treated with ultrasonic waves differed in appearance from those treated with NaOCl; the collagen fibrils were thin and displayed a smooth surface in type I lacunae treated with ultrasonic waves, while they were thick and showed a rough surface in those treated with NaOCl-probably because superficial uncalcified collagen fibrils were digested with the chemical. The results indicated that type I lacunae occupied 77% of all of the bone resorption lacunae treated with ultrasonic waves, but 51% of those treated with NaOCl. This finding led to the idea that type I lacunae can be subdivided into two: lacunae (Ia), covered with partially calcified fibrils as well as superficial uncalcified fibrils; and lacunae (Ib), covered only with uncalcified fibrils. The presence of uncalcified fibrils in the bone resorption lacunae was further confirmed by backscattered electron (BSE) imaging of SEM. Histochemistry for acid phosphatase or immuno-histochemistry for cathepsin B or carbonic anhydrase in combination with SEM revealed that type I lacunae were located under osteoclasts but type II lacunae were not. These findings indicate that type I lacunae are in the process of bone resorption by osteoclasts, while type II lacunae are in the final stage of bone resorption and free from osteoclasts. Bone resorption may thus proceed in the order of Ia, Ib, and II. PMID:16079456

Ren, Shumei; Takano, Hiroko; Abe, Kazuhiro

2005-06-01

201

Synaptic Properties of Corticocortical Connections between the Primary and Secondary Visual Cortical Areas in the Mouse  

PubMed Central

Despite the importance of corticocortical connections, few published studies have investigated the functional, synaptic properties of such connections in any species, because most studies have been purely anatomical or aimed at functional features other than synaptic properties. We recently published a study of synaptic properties of connections between the primary and secondary cortical auditory areas in brain slices from the mouse, and in the present study we aimed to extend this by performing analogous studies of the primary and secondary visual areas (V1 and V2). We found effectively the same results. That is, connections between V1 and V2 in both directions were quite similar; in each case the glutamatergic inputs could be classified as one of two types, Classes 1B (formerly “driver”) and 2 (formerly “modulator”). There is a clear laminar correlation for these different inputs, both in terms of the laminae of origin and those in which the recorded cells were located. Our data suggest common pattern to the functional organization of corticocortical connectivity in the mouse cortex.

De Pasquale, Roberto; Sherman, S. Murray

2011-01-01

202

Establishment of primary cultures for mouse ameloblasts as a model of their lifetime  

SciTech Connect

To understand how the properties of ameloblasts are spatiotemporally regulated during amelogenesis, two primary cultures of ameloblasts in different stages of differentiation were established from mouse enamel epithelium. Mouse primary ameloblasts (MPAs) prepared from immature enamel epithelium (MPA-I) could proliferate, whereas those from mature enamel epithelium (MPA-M) could not. MPA-M but not MPA-I caused apoptosis during culture. The mRNA expression of amelogenin, a marker of immature ameloblasts, was down-regulated, and that of enamel matrix serine proteiase-1, a marker of mature ameloblasts, was induced in MPA-I during culture. Using green fluorescence protein as a reporter, a visualized reporter system was established to analyze the promoter activity of the amelogenin gene. The region between -1102 bp and -261 bp was required for the reporter expression in MPA-I. These results suggest that MPAs are valuable in vitro models for investigation of ameloblast biology, and that the visualized system is useful for promoter analysis in MPAs.

Suzawa, Tetsuo [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan)]. E-mail: suzawa@dent.showa-u.ac.jp; Itoh, Nao [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan); Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Takahashi, Naoyuki [Institute for Oral Science, Matsumoto Dental University, Shiojiri 399-0781 (Japan); Katagiri, Takenobu [Division of Pathophysiology, Research Center for Genomic Medicine, Saitama Medical School, Hidaka 350-1241 (Japan); Morimura, Naoko [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan); Kobayashi, Yasuna [Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Yamamoto, Toshinori [Department of Clinical Pharmacy, School of Pharmaceutical Science, Showa University (Japan); Kamijo, Ryutaro [Department of Biochemistry, School of Dentistry, Showa University, Tokyo 142-8555 (Japan)

2006-07-07

203

Endogenous retrovirus induces leukemia in a xenograft mouse model for primary myelofibrosis.  

PubMed

The compound immunodeficiencies in nonobese diabetic (NOD) inbred mice homozygous for the Prkdc(scid) and Il2rg(null) alleles (NSG mice) permit engraftment of a wide-range of primary human cells, enabling sophisticated modeling of human disease. In studies designed to define neoplastic stem cells of primary myelofibrosis (PMF), a myeloproliferative neoplasm characterized by profound disruption of the hematopoietic microenvironment, we observed a high frequency of acute myeloid leukemia (AML) in NSG mice. AML was of mouse origin, confined to PMF-xenografted mice, and contained multiple clonal integrations of ecotropic murine leukemia virus (E-MuLV). Significantly, MuLV replication was not only observed in diseased mice, but also in nontreated NSG controls. Furthermore, in addition to the single ecotropic endogenous retrovirus (eERV) located on chromosome 11 (Emv30) in the NOD genome, multiple de novo germ-line eERV integrations were observed in mice from each of four independent NSG mouse colonies. Analysis confirmed that E-MuLV originated from the Emv30 provirus and that recombination events were not necessary for virus replication or AML induction. Pathogenicity is thus likely attributable to PMF-mediated paracrine stimulation of mouse myeloid cells, which serve as targets for retroviral infection and transformation, as evidenced by integration into the Evi1 locus, a hotspot for retroviral-induced myeloid leukemia. This study thus corroborates a role of paracrine stimulation in PMF disease progression, underlines the importance of target cell type and numbers in MuLV-induced disease, and mandates awareness of replicating MuLV in NOD immunodeficient mice, which can significantly influence experimental results and their interpretation. PMID:24912157

Triviai, Ioanna; Ziegler, Marion; Bergholz, Ulla; Oler, Andrew J; Stübig, Thomas; Prassolov, Vladimir; Fehse, Boris; Kozak, Christine A; Kröger, Nicolaus; Stocking, Carol

2014-06-10

204

Progesterone Receptor-Induced Gene Expression in Primary Mouse Granulosa Cell Cultures1  

PubMed Central

The progesterone receptor (PGR) is induced by luteinizing hormone (LH) in granulosa cells of preovulatory follicles, and the PGR-A isoform is essential for ovulation based on the phenotypes of Pgr isoform-specific knockout mice. Although several genes regulated by PGR-A in vivo have been identified, whether these genes are primary targets of PGR-A or if their expression also depends on other signaling molecules that are induced by the LH surge has not been resolved. Therefore, to identify genes that are either induced or repressed by PGR in the absence of LH-mediated signaling cascades, we infected primary cultures of mouse granulosa cells with either PGR-A or PGR-B adenoviral vectors without or with R-5020 as a PGR ligand. Total RNA was extracted from infected cells at 16 h and analyzed by Affymetrix Mouse 430 2.0 microarrays. PGR-A in the presence or absence of ligand significantly induced approximately 50 genes 2-fold or more (local pooled error test at P ? 0.01). Fewer and different genes were induced by PGR-B in the absence of ligand. Edn1, Apoa1, and Cited1 were primarily regulated by PGR-A as verified by additional RT-PCR analyses, suppression by the PGR antagonist RU486, and the lack of induction by protein kinase A, protein kinase C, or epidermal growth factor (EGF)-like factors pathways. PGR regulation of these genes was confirmed further by gene expression analyses in hormonally primed Pgr mutant mouse ovaries. Because Edn1, Apoa1, and Cited1 are known to regulate angiogenesis, PGR may affect the neovascularization of follicles that is initiated with ovulation.

Sriraman, Venkataraman; Sinha, Mala; Richards, JoAnne S.

2009-01-01

205

Examination of ER? Signaling Pathways in Bone of Mutant Mouse Models Reveals the Importance of ERE-Dependent Signaling  

PubMed Central

The mechanisms of estrogen receptor (ER)-? activity can be categorized into those involving direct (classical) or indirect (nonclassical) DNA binding. Although various mouse models have demonstrated the importance of ER? in bone, the specific gene expression patterns affected by these modes of ER? action are unknown. In this report, the gene expression patterns of ER?-deficient (ERKO) mice and nonclassical ER knock-in (NERKI) mice, which can function only by nonclassical means, were analyzed. Three-month-old mice were ovariectomized and implanted with estrogen pellets for 1 month to normalize estrogen levels. Microarray analysis of flushed cortical bone revealed 28% (210 of 763) of the genes differentially expressed in ERKO mice were altered in NERKI mice, suggesting estrogen response element-dependent regulation of these genes in bone. Pathway analysis revealed alterations in genes involved in focal adhesion and extracellular matrix interactions. However, the majority of genes regulated in ERKO mice (72%) were unique (i.e. not altered in NERKI mice), suggesting these are regulated by nonclassical mechanisms. To further explore the pathways affected in ERKO mice, we performed focused quantitative PCR arrays for genes involved in various aspects of bone physiology. Genes involved in bone formation, senescence, apoptosis, and autophagy were significantly regulated. Overall, the majority of the genes regulated by ER? in bone are via nonclassical pathways. However, because NERKI mice display an osteoporotic phenotype, it can be deduced that the minority of the estrogen response element-dependent genes/pathways play critical roles in the regulation of bone physiology. These data demonstrate the importance of classical ER? signaling in regulating bone metabolism.

Chokalingam, Kumar; Roforth, Matthew M.; Nicks, Kristy M.; McGregor, Ulrike; Fraser, Daniel; Khosla, Sundeep

2012-01-01

206

Levels and distribution of organochlorine pollutants in primary dental tissues and bone of lamb.  

PubMed

This study examined the bioconcentration of selected organochlorine pollutants, tetra- and hexa-chlorobiphenyls with planar (PCB-80, PCB-169) and non-planar (PCB-54, PCB-155) structure, and persistent organochlorine pesticides with planar [hexachlorobenzene (HCB)] and non-planar [1,1-bis (4-chlorophenyl)-2,2-dichloroethene (4,4'-DDE)] structure in primary dental tissues (pulp, dentine, and enamel) and mandibular bone of lactationally exposed lambs, and compared it with the organochlorines distribution pattern in permanent dental tissues and bone. Also, the role of pollutants physicochemical properties and tissue specific characteristics in the bioconcentration was assessed. Residual levels of individual pollutants were analyzed by high-resolution gas chromatography with electron-capture detection. Our results showed that transfer of organochlorines to primary hard dental tissues was higher than to permanent hard dental tissues. Metabolically more stable, planar, and toxic organochlorines (e.g. PCB-169 and HCB) predominated in primary hard dental tissues, where they may represent a potential risk for developmental dental defects. PMID:24100271

Jan, Janja; Urši?, Matjaž; Vrecl, Milka

2013-11-01

207

Increased bone mass is a part of the generalized lymphoproliferative disorder phenotype in the mouse.  

PubMed

We investigated the bone phenotype of mice with generalized lymphoproliferative disorder (gld) due to a defect in the Fas ligand-mediated apoptotic pathway. C57BL/6-gld mice had greater whole body bone mineral density and greater trabecular bone volume than their wild-type controls. gld mice lost 5-fold less trabecular bone and had less osteoclasts on bone surfaces after ovariectomy-induced bone resorption. They also formed more bone in a model of osteogenic regeneration after bone marrow ablation, had less osteoclasts on bone surfaces and less apoptotic osteoblasts. gld and wild-type mice had similar numbers of osteoclasts in bone marrow cultures, but marrow stromal fibroblasts from gld mice formed more alkaline phosphatase-positive colonies. Bone diaphyseal shafts and bone marrow stromal fibroblasts produced more osteoprotegerin mRNA and protein than wild-type mice. These findings provide evidence that the disturbance of the bone system is a part of generalized lymphoproliferative syndrome and indicates the possible role of osteoprotegerin as a regulatory link between the bone and immune system. PMID:12538719

Katavi?, Vedran; Luki?, Ivan Kresimir; Kovaci?, Natasa; Grcevi?, Danka; Lorenzo, Joseph A; Marusi?, Ana

2003-02-01

208

Effects of swimming and weight loading on bone density and mechanical properties of the mouse femoral bone  

Microsoft Academic Search

Effects of swimming and weight loading on femoral bone density and mechanical properties of the bone were investigated. ICR\\u000a 8-week old female mice were divided into a swimming, weight-loading and control groups consisting 10 mice in each group. All\\u000a mice were fed a standard diet and waterad libitum during the entire experiment. Body weight was significantly lower in the swimming

Akio Hoshi; Hiromi Watanabe; Momoko Chiba; Yutaka Inaba

1996-01-01

209

[Efficient isolation of mesenchymal stem cells from human bone marrow by direct plating method combined with modified primary explant culture].  

PubMed

Human bone marrow is the major source of mesenchymal stem cells (MSC). It was reported that the standard density gradient centrifugation method was not efficient in isolating MSC and it may be caused by the existing of bone marrow particles. In previous study, a lot of MSC were obtained by culturing bone marrow particles alone combined with standard method. However, it is time- and labor-consuming to obtain bone marrow particles by filtering and to isolate MNC by density gradient centrifugation. This study was purposed to explore the more simple and efficient method to isolate MSC from bone marrow. Seven normal bone marrow aspirates were collected and centrifugated. The bone marrow particles floated on surface layers were cultured by modified primary explant culture, whereas the bone marrow aspirates deposited were cultured by direct plating method, then the immun phenotype and differentiation capability of isolated cells were analyzed. The results showed that in 3 of 7 aspirates, bone marrow particles were floated on surface layers, whereas the other bone marrow cells and some particles were deposited after centrifugation. The MSC were reliably isolated from the floating layers or deposited aspirates by modified primary explant culture and direct plating method separately. After 3 passages the isolated MSC did not express CD45 and CD34, but expressed CD105, CD73, CD44, CD90, CD49e and they could differentiate into chondrocytes and adipocytes. It is concluded that normal human bone marrow MSC can be isolated simply and efficiently by direct plating method in combination with modified primary explant culture. PMID:23628052

Xing, Wen; Pang, Ai-Ming; Yao, Jian-Feng; Li, Yuan; Shi, Hui; Sheng, Meng-Yao; Zhou, Yuan; Zhao, Ying-Xu; Xu, Ming-Jiang; Yang, Feng-Chun

2013-04-01

210

Longitudinal evaluation of mouse hind limb bone loss after spinal cord injury using novel, in vivo, methodology.  

PubMed

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher frequency of fractures. As these fractures often occur in regions that have lost normal sensory function, the patient is at a greater risk of fracture-dependent pathologies, including death. SCI-dependent loss in both bone mineral density (BMD, grams/cm2) and bone mineral content (BMC, grams) has been attributed to mechanical disuse, aberrant neuronal signaling and hormonal changes. The use of rodent models of SCI-induced osteoporosis can provide invaluable information regarding the mechanisms underlying the development of osteoporosis following SCI as well as a test environment for the generation of new therapies. Mouse models of SCI are of great interest as they permit a reductionist approach to mechanism-based assessment through the use of null and transgenic mice. While such models have provided important data, there is still a need for minimally-invasive, reliable, reproducible, and quantifiable methods in determining the extent of bone loss following SCI, particularly over time and within the same cohort of experimental animals, to improve diagnosis, treatment methods, and/or prevention of SCI-induced osteoporosis. An ideal method for measuring bone density in rodents would allow multiple, sequential (over time) exposures to low-levels of X-ray radiation. This study describes the use of a new whole-animal scanner, the IVIS Lumina XR (Caliper Instruments) that can be used to provide low-energy (1-3 milligray (mGy)) high-resolution, high-magnification X-ray images of mouse hind limb bones over time following SCI. Significant bone density loss was seen in the tibiae of mice by 10 days post-spinal transection when compared to uninjured, age-matched control (naïve) mice (13% decrease, p < 0.0005). Loss of bone density in the distal femur was also detectable by day 10 post-SCI, while a loss of density in the proximal femur was not detectable until 40 days post injury (7% decrease, p < 0.05). SCI-dependent loss of mouse femur density was confirmed post-mortem through the use of Dual-energy X-ray Absorptiometry (DXA), the current "gold standard" for bone density measurements. We detect a 12% loss of BMC in the femurs of mice at 40 days post-SCI using the IVIS Lumina XR. This compares favorably with a previously reported BMC loss of 13.5% by Picard and colleagues who used DXA analysis on mouse femurs post-mortem 30 days post-SCI (9). Our results suggest that the IVIS Lumina XR provides a novel, high-resolution/high-magnification method for performing long-term, longitudinal measurements of hind limb bone density in the mouse following SCI. PMID:22158515

McManus, Madonna M; Grill, Raymond J

2011-01-01

211

The role of bone marrow-derived cells in bone fracture repair in a green fluorescent protein chimeric mouse model  

SciTech Connect

We investigated the role of bone marrow cells in bone fracture repair using green fluorescent protein (GFP) chimeric model mice. First, the chimeric model mice were created: bone marrow cells from GFP-transgenic C57BL/6 mice were injected into the tail veins of recipient wild-type C57BL/6 mice that had been irradiated with a lethal dose of 10 Gy from a cesium source. Next, bone fracture models were created from these mice: closed transverse fractures of the left femur were produced using a specially designed device. One, three, and five weeks later, fracture lesions were extirpated for histological and immunohistochemical analyses. In the specimens collected 3 and 5 weeks after operation, we confirmed calluses showing intramembranous ossification peripheral to the fracture site. The calluses consisted of GFP- and osteocalcin-positive cells at the same site, although the femur consisted of only osteocalcin-positive cells. We suggest that bone marrow cells migrated outside of the bone marrow and differentiated into osteoblasts to make up the calluses.

Taguchi, Kazuhiro [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan) and Department of Orthopedic Surgery, Nippon Medical School, Tokyo (Japan)]. E-mail: s3061@nms.ac.jp; Ogawa, Rei [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan); Department of Plastic and Reconstructive Surgery, Nippon Medical School, Tokyo (Japan); Migita, Makoto [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan); Department of Pediatrics, Nippon Medical School, Tokyo (Japan); Hanawa, Hideki [Department of Plastic and Reconstructive Surgery, Nippon Medical School, Tokyo (Japan); Ito, Hiromoto [Department of Orthopedic Surgery, Nippon Medical School, Tokyo (Japan); Orimo, Hideo [Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo (Japan)

2005-05-27

212

Positive selection in bone morphogenetic protein 15 targets a natural mutation associated with primary ovarian insufficiency in human.  

PubMed

Bone Morphogenetic Protein 15 (BMP15) is a TGF?-like oocyte-derived growth factor involved in ovarian folliculogenesis as a critical regulator of many granulosa cell processes. Alterations of the BMP15 gene have been found associated with different ovarian phenotypic effects depending on the species, from sterility to increased prolificacy in sheep, slight subfertility in mouse or associated with primary ovarian insufficiency (POI) in women. To investigate the evolving role of BMP15, a phylogenetic analysis of this particular TGF? family member was performed. A maximum likelihood phylogenetic tree of several TGF?/BMP family members expressed by the ovary showed that BMP15 has a very strong divergence and a rapid evolution compared to others. Moreover, among 24 mammalian species, we detected signals of positive selection in the hominidae clade corresponding to F146, L189 and Y235 residues in human BMP15. The biological importance of these residues was tested functionally after site directed-mutagenesis in a COV434 cells luciferase assay. By replacing the positively selected amino acid either by alanine or the most represented residue in other studied species, only L189A, Y235A and Y235C mutants showed a significant increase of BMP15 signaling when compared to wild type. Additionally, the Y235C mutant was more potent than wild type in inhibiting progesterone secretion of ovine granulosa cells in primary culture. Interestingly, the Y235C mutation was previously identified in association with POI in women. In conclusion, this study evidences that the BMP15 gene has evolved faster than other members of the TGFß family and was submitted to a positive selection pressure in the hominidae clade. Some residues under positive selection are of great importance for the normal function of the protein and thus for female fertility. Y235 represents a critical residue in the determination of BMP15 biological activity, thus indirectly confirming its role in the onset of POI in women. PMID:24147118

Auclair, Sylvain; Rossetti, Raffaella; Meslin, Camille; Monestier, Olivier; Di Pasquale, Elisa; Pascal, Géraldine; Persani, Luca; Fabre, Stéphane

2013-01-01

213

Gene expression of bone morphogenic protein 8B in the primary site, peripheral blood and bone marrow of patients with gastric cancer  

PubMed Central

The prognosis for individuals that are diagnosed with gastric cancer remains poor due to the high frequency of metastatic disease. In response to tumor-derived secreted factors, the bone marrow generates a suitable microenvironment for the development of metastasis. However, it is largely unknown whether secreted factors in bone marrow associated with metastatic disease of patients with gastric cancer are present. Secreted factors from the bone marrow of patients with metastatic gastric cancer were identified using a DNA microarray analysis and the mRNA expression levels were investigated in 355 bone marrow, 295 peripheral blood and 144 primary site samples using quantitative PCR (qPCR). Using DNA microarray analysis, the present study identified bone morphogenetic protein 8B (BMP8B) as a secreted signaling molecule in the bone marrow that was associated with the metastatic disease of human gastric cancer. The expression levels of BMP8B in the bone marrow of 355 gastric cancer patients were increased with metastatic disease. A significant correlation was demonstrated between BMP8B mRNA expression in the bone marrow and in the peripheral blood. High BMP8B expression in the bone marrow was associated with the diffuse type of gastric cancer (P=0.009), lymph node metastasis (P=0.009), liver metastasis (P=0.044) and peritoneal dissemination (P<0.001). In the primary site, a multivariate analysis revealed BMP8B mRNA expression as one of the independent prognostic factors of gastric cancer [hazard ratio (HR), 2.066; 95% CI, 1.132–3.772]. This study suggests that BMP8B, a previously unknown secreted factor in cancer progression, has the potential to be used as a prognostic biomarker. The present study may provide insight into a new mechanism that underlies the dissemination of gastric cancer cells.

MIMA, KOSUKE; FUKAGAWA, TAKEO; KURASHIGE, JUNJI; TAKANO, YUKI; UCHI, RYUTARO; UEO, HIROKI; MATSUMURA, TAE; ISHIBASHI, MASAHISA; SAWADA, GENTA; TAKAHASHI, YUSUKE; AKIYOSHI, SAYURI; EGUCHI, HIDETOSHI; SUDO, TOMOYA; SUGIMACHI, KEISHI; WATANABE, MASAYUKI; ISHII, HIDESHI; MORI, MASAKI; BABA, HIDEO; SASAKO, MITSURU; MIMORI, KOSHI

2013-01-01

214

Constructing a multi-scan synchrotron X-ray microscope to study the function of osteocyte canaliculi in mouse bone  

SciTech Connect

Formulating a multi-scan method applied to an X-ray microscope CT with synchrotron radiation, we attempted to analyze the 3D functional structure of osteocyte canaliculi inside the cortical bone of a mouse tibia. We employed a two-method combination to scan the same position of the specimen. To extract the internal bone canalicular structure, we first combined a Talbot interferometer with an X-ray microscope, and applied a differential phase imaging method to measure the absolute value of bone mineral around the canaliculi. Next, we used the X-ray microscope without the Talbot interferometer under a defocus condition, moving the specimen toward the zone plate by 6 mm. This defocus contrast method visualizes the canaliculi by emphasizing the edges of the bone. We performed CT scans by the two configurations and precisely aligned resultant 3D images so that the same position in the specimen is compared. We could extract the osteocyte canaliculi and evaluate the mineral density of their surroundings. The degree of mineralization varied for each osteocyte lacuna and canaliculus. The multi-scan microscopic X-ray CT is a powerful tool for analyzing bone mineralization.

Nango, Nobuhito; Kubota, Shogo; Yashiro, Wataru; Momose, Atsushi; Takada, Yasunari; Matsuo, Koichi [Ratoc System Engineering Co., Ltd, Toho Edogawabashi Bldg. 4F, 1-24-8 Sekiguchi, Bunkyo-ku, Tokyo 112-0014 (Japan); Dept. of Advanced Materials Science, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa-shi, Chiba 277-8561 (Japan); Lab. of Cell and Tissue Biology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan)

2012-07-31

215

Enumeration of the colony-forming units-fibroblast from mouse and human bone marrow in normal and pathological conditions  

PubMed Central

Bone marrow stromal cell populations, containing a subset of multipotential skeletal stem cells, are increasingly contemplated for use in tissue engineering and stem cell therapy, whereas their involvement in the pathogenetic mechanisms of skeletal disorders is far less recognized. We compared the concentrations of stromal clonogenic cells, colony forming units–fibroblast (CFU-Fs), in norm and pathology. Initially, culture conditions were optimized by demonstrating that fetal bovine serum heat inactivation could significantly repress colony formation. Using non-heat-inactivated fetal bovine serum, the concentration of CFU-Fs (colony-forming efficiency, CFE) ranged from 3.5 ± 1.0 to 11.5 ± 4.0 per 1 × 105 nucleated cells in five inbred mouse strains. In four transgenic lines with profound bone involvement, CFE was either significantly reduced or increased compared to wild-type littermates. In normal human donors, CFE decreased slightly with age and averaged 52.2 ± 4.1 for children and 32.3 ± 3.0 for adults. CFE was significantly altered in patients with several skeletal, metabolic, and hematological disorders: reduced in congenital generalized lipodystrophy, achondroplasia (SADDAN), pseudoachondroplasia, and Paget disease of bone and elevated in alcaptonuria and sickle cell anemia. Our findings indicate that under appropriate culture conditions, CFE values may provide useful insights into bone/bone marrow pathophysiology.

Kuznetsov, Sergei A.; Mankani, Mahesh H.; Bianco, Paolo; Robey, Pamela G.

2009-01-01

216

Constructing a multi-scan synchrotron X-ray microscope to study the function of osteocyte canaliculi in mouse bone  

NASA Astrophysics Data System (ADS)

Formulating a multi-scan method applied to an X-ray microscope CT with synchrotron radiation, we attempted to analyze the 3D functional structure of osteocyte canaliculi inside the cortical bone of a mouse tibia. We employed a two-method combination to scan the same position of the specimen. To extract the internal bone canalicular structure, we first combined a Talbot interferometer with an X-ray microscope, and applied a differential phase imaging method to measure the absolute value of bone mineral around the canaliculi. Next, we used the X-ray microscope without the Talbot interferometer under a defocus condition, moving the specimen toward the zone plate by 6 mm. This defocus contrast method visualizes the canaliculi by emphasizing the edges of the bone. We performed CT scans by the two configurations and precisely aligned resultant 3D images so that the same position in the specimen is compared. We could extract the osteocyte canaliculi and evaluate the mineral density of their surroundings. The degree of mineralization varied for each osteocyte lacuna and canaliculus. The multi-scan microscopic X-ray CT is a powerful tool for analyzing bone mineralization.

Nango, Nobuhito; Kubota, Shogo; Yashiro, Wataru; Momose, Atsushi; Takada, Yasunari; Matsuo, Koichi

2012-07-01

217

Bone marrow stromal cells as an inducer for cardiomyocyte differentiation from mouse embryonic stem cells.  

PubMed

Bone marrow stromal cells (BMSCs) secrete soluble factors and display varied cell-biological functions. To confirm the ability and efficiency of BMSCs to induce embryonic stem cells (ESCs) into cardiomyocytes, mouse embryoid bodies (EBs) were co-cultured with rat BMSCs. After about 10 days, areas of rhythmically contracting cells in more solid aggregates became evident with bundle-like structures formed along borders between EB outgrowth and BMSC layer. ESC-derived cardiomyocytes exhibited sarcomeric striations when stained with troponin I (Trop I), organized in separated bundles. Besides, the staining for connexin 43 was detected in cell-cell junctions, which demonstrated that ESC-derived cardiomyocytes were coupled by gap junction in culture. The related genes of cardiomyocytes were found in these beating and no-beating EBs co-cultured with BMSCs. In addition, an improved efficiency of cardiomyocyte differentiation from ESC-BMSC co-culture was found in the serum-free medium: 5-fold up-regulation in the number of beating area compared with the serum medium. Effective cardiac differentiation was also recognized in transfer filter assay and in condition medium obtained from BMSC culture. A clear increase in the expression of cardiac genes and TropI protein confirmed further cardiac differentiation by BMP4 and Retinoic Acid (RA) treatment. These results demonstrate that BMSCs can induce cardiomyocyte differentiation from ESCs through soluble factors and enhance it with BMP4 or RA treatment. Serum-free ESC-BMSC co-culture represents a defined in vitro model for identifying the cardiomyocyte-inducing activity from BMSCs and, in addition, a straightforward experimental system for assessing clinical applications. PMID:20801009

Yue, Fengming; Johkura, Kohei; Tomotsune, Daihachiro; Shirasawa, Sakiko; Yokoyama, Tadayuki; Nagai, Mika; Sasaki, Katsunori

2010-09-20

218

Allogeneic bone marrow transplantation from partially mismatched related donors as therapy for primary induction failure acute myeloid leukemia  

Microsoft Academic Search

The outcome of acute myeloid leukemia patients with primary refractoriness to conventional chemotherapy is extremely poor. Allogeneic bone marrow transplants with matched sibling or matched unrelated donors provide 10–20% disease-free survival in this setting. We analyzed our transplant experience using readily available partially mismatched related donor (PMRD) in patients with primary induction failure (PIF) AML. Between March 1994 and December

KY Chiang; F Van Rhee; K Godder; K Bridges; S Adams; J Mehta; PJ Henslee-Downey

2001-01-01

219

Tissue Preparation and Immunostaining of Mouse Sensory Nerve Fibers Innervating Skin and Limb Bones  

PubMed Central

Detection and primary processing of physical, chemical and thermal sensory stimuli by peripheral sensory nerve fibers is key to sensory perception in animals and humans. These peripheral sensory nerve fibers express a plethora of receptors and ion channel proteins which detect and initiate specific sensory stimuli. Methods are available to characterize the electrical properties of peripheral sensory nerve fibers innervating the skin, which can also be utilized to identify the functional expression of specific ion channel proteins in these fibers. However, similar electrophysiological methods are not available (and are also difficult to develop) for the detection of the functional expression of receptors and ion channel proteins in peripheral sensory nerve fibers innervating other visceral organs, including the most challenging tissues such as bone. Moreover, such electrophysiological methods cannot be utilized to determine the expression of non-excitable proteins in peripheral sensory nerve fibers. Therefore, immunostaining of peripheral/visceral tissue samples for sensory nerve fivers provides the best possible way to determine the expression of specific proteins of interest in these nerve fibers. So far, most of the protein expression studies in sensory neurons have utilized immunostaining procedures in sensory ganglia, where the information is limited to the expression of specific proteins in the cell body of specific types or subsets of sensory neurons. Here we report detailed methods/protocols for the preparation of peripheral/visceral tissue samples for immunostaining of peripheral sensory nerve fibers. We specifically detail methods for the preparation of skin or plantar punch biopsy and bone (femur) sections from mice for immunostaining of peripheral sensory nerve fibers. These methods are not only key to the qualitative determination of protein expression in peripheral sensory neurons, but also provide a quantitative assay method for determining changes in protein expression levels in specific types or subsets of sensory fibers, as well as for determining the morphological and/or anatomical changes in the number and density of sensory fibers during various pathological states. Further, these methods are not confined to the staining of only sensory nerve fibers, but can also be used for staining any types of nerve fibers in the skin, bones and other visceral tissue.

Shepherd, Andrew J.; Mohapatra, Durga P.

2012-01-01

220

Skeletal effects of primary hyperparathyroidism: bone mineral density and fracture risk.  

PubMed

Parathyroid hormone (PTH) is associated with anabolic and catabolic skeletal effects that vary according to the kinetics of serum levels and the type of bone. The anabolic effects are manifested in patients with a periodic rapid transient rise in serum PTH, as seen with daily subcutaneous injection of PTH(1-34) and PTH(1-84) in the treatment of osteoporosis. These patients have an increase in bone mineral density (BMD), particularly at skeletal sites with a high trabecular component, such as the lumbar spine, and a reduction in fracture risk. The catabolic effects are typified in patients with primary hyperparathyroidism (PHPT) who have chronic persistently elevated PTH levels. Patients with long-standing PHPT have a reduction in BMD, particularly at predominately cortical skeletal sites, such as the one-third radius, with relative preservation of BMD at the lumbar spine. Some but not all studies have reported an increase in fracture risk with PHPT. Because many patients with PHPT are postmenopausal women at risk for osteoporosis owing to estrogen deficiency, BMD and fracture risk may be a result of multiple factors with variable effects on bone remodeling. The skeletal effects of normocalcemic PHPT have not yet been fully characterized, but may not be the same as hypercalcemic PHPT. PMID:23374738

Lewiecki, E Michael; Miller, Paul D

2013-01-01

221

Nonhematopoietic Cells are the Primary Source of Bone Marrow-Derived Lung Epithelial Cells  

PubMed Central

Previous studies have demonstrated that bone marrow (BM)-derived cells differentiate into nonhematopoietic cells of multiple tissues. To date, it remains unknown which population(s) of BM cells are primarily responsible for this engraftment. To test the hypothesis that nonhematopoietic stem cells in the BM are the primary source of marrow-derived lung epithelial cells, either wild-type hematopoietic or nonhematopoietic BM cells were transplanted into irradiated surfactant-protein-C (SPC)-null mice. Donor-derived, SPC-positive type 2 pneumocytes were predominantly detected in the lungs of mice receiving purified nonhematopoietic cells and were absent from mice receiving purified hematopoietic stem and progenitor cells. We conclude that cells contained in the nonhematopoietic fraction of the BM are the primary source of marrow-derived lung epithelial cells. These nonhematopoietic cells may represent a primitive stem cell population residing in adult BM.

Kassmer, Susannah H.; Bruscia, Emanuela M.; Zhang, Ping-Xia; Krause, Diane S.

2013-01-01

222

A mouse bone marrow stromal cell line with skeletal stem cell characteristics to study osteogenesis in vitro and in vivo.  

PubMed

Bone marrow stromal cells (BMSCs) are composed of progenitor and multipotent skeletal stem cells, which are able to differentiate in vitro into osteocytes, adipocytes, and chondrocytes. Mouse BMSCs (mBMSCs) are a versatile model system to investigate factors involved in BMSC differentiation in vitro and in vivo as a variety of transgenic mouse models are available. In this study, mBMSCs were isolated and osteogenic differentiation was investigated in tissue culture and in vivo. Three out of seven independent cell isolates showed the ability to differentiate into osteocytes, adipocytes, and chondrocytes in vitro. In vitro multipotency of an established mBMSC line was maintained over 45 passages. The osteogenic differentiation of this cell line was confirmed by quantitative polymerase chain reaction (qPCR) analysis of specific markers such as osteocalcin and shown to be Runx2 dependent. Notably, the cell line, when transplanted subcutaneously into mice, possesses full skeletal stem cell characteristics in vivo in early and late passages, evident from bone tissue formation, induction of vascularization, and hematopoiesis. This cell line provides, thus, a versatile tool to unravel the molecular mechanisms governing osteogenesis in vivo thereby aiding to improve current strategies in bone regenerative therapy. PMID:24405418

Raeth, Sebastian; Sacchetti, Benedetto; Siegel, Georg; Mau-Holzmann, Ulrike A; Hansmann, Jan; Vacun, Gabriele; Hauk, Thomas G; Pfizenmaier, Klaus; Hausser, Angelika

2014-05-15

223

Long Term Cyclic Pamidronate Reduces Bone Growth by Inhibiting Osteoclast Mediated Cartilage-to-Bone Turnover in the Mouse  

PubMed Central

Bisphosphonates, used to treat diseases exhibiting increased osteoclast activity, reduce longitudinal bone growth through an as yet undefined mechanism. Pamidronate, an aminobisphosphonate, was given weekly to mice at 0, 1.25, or 2.50 mg/kg/wk beginning at 4 weeks of age. At 12 weeks of age, humeral length, growth plate area, regional chondrocyte cell numbers, chondrocyte apoptosis, TRAP stained osteoclast number, and osteoclast function assessed by cathepsin K immunohistochemistry were quantified. Humeral length was decreased in pamidronate treated mice compared to vehicle control mice, and correlated with greater growth plate areas reflecting greater proliferative and hypertrophic chondrocyte cell numbers with fewer hypertrophic cells undergoing apoptosis. Pamidronate treatment increased TRAP stained osteoclast numbers yet decreased cathepsin K indicating that pamidronate repressed osteoclast maturation and function. The data suggest that long term cyclic pamidronate treatment impairs bone growth by inhibition of osteoclast maturation thereby reducing cartilage-to-bone turnover within the growth plate.

Evans, K.D; Sheppard, L.E; Grossman, D.I; Rao, S.H; Martin, R.B; Oberbauer, A.M

2008-01-01

224

On the relation between surface roughness of metallic substrates and adhesion of human primary bone cells.  

PubMed

Surface characteristics of materials, whether their topography, chemistry, or surface energy, play an essential part in osteoblast adhesion on biomaterials. Thus, the quality of cell adhesion will influence the cell's capacity to proliferate and differentiate in contact with a biomaterial. We have developed for more than ten years numerous studies on the influence of topography and chemistry of metallic substrates on the response of primary human bone cells. The originality of our approach is that contrary to most of other authors, we quantified the adhesion of primary human bone cells on metallic substrates with perfectly characterized surface topography after some hours but also over 21 days. Moreover, we have developed original statistical approaches for characterizing the relation between surface roughness and cell-adhesion parameters. In this article, we will illustrate different studies we did these last ten years concerning the development of a new adhesion parameter, the adhesion power; the correlation between short-term adhesion, long-term adhesion, and proliferation; the influence of roughness organization on cell adhesion and the development of the order parameter; our modeling approach of cell adhesion on surface topography; the relative influence of surface chemistry and topography on cell adhesion and contact angle; the relation between surface features dimensions and cell adhesion. Further, some considerations will be given on the methods for scanning surface topography for cell-adhesion studies. Finally, perspectives will be given to elucidate these intracellular mechanotransduction mechanisms induced by the deformation of cells on model sinusoidal peaks-or-valleys surfaces. PMID:23203601

Anselme, K; Bigerelle, M

2014-01-01

225

Endoprosthetic reconstruction of the distal tibia and ankle joint after resection of primary bone tumours.  

PubMed

Endoprosthetic replacement of the distal tibia and ankle joint for a primary bone tumour is a rarely attempted and technically challenging procedure. We report the outcome of six patients treated between 1981 and 2007. There were four males and two females, with a mean age of 43.5 years (15 to 75), and a mean follow-up of 9.6 years (1 to 27). No patient developed a local recurrence or metastasis. Two of the six went on to have a below-knee amputation for persistent infection after a mean 16 months (1 to 31). The four patients who retained their endoprosthesis had a mean musculoskeletal tumour society score of 70% and a mean Toronto extremity salvage score of 71%. All were pain free and able to perform most activities of daily living in comfort. A custom-made endoprosthetic replacement of the distal tibia and ankle joint is a viable treatment option for carefully selected patients with a primary bone tumour. Patients should, however, be informed of the risk of infection and the potential need for amputation if this cannot be controlled. PMID:19794176

Shekkeris, A S; Hanna, S A; Sewell, M D; Spiegelberg, B G I; Aston, W J S; Blunn, G W; Cannon, S R; Briggs, T W R

2009-10-01

226

Organization of Estrogen-Associated Circuits in the Mouse Primary Auditory Cortex.  

PubMed

Sex steroid hormones influence the perceptual processing of sensory signals in vertebrates. In particular, decades of research have shown that circulating levels of estrogen correlate with hearing function. The mechanisms and sites of action supporting this sensory-neuroendocrine modulation, however, remain unknown. Here we combined a molecular cloning strategy, fluorescence in-situ hybridization and unbiased quantification methods to show that estrogen-producing and -sensitive neurons heavily populate the adult mouse primary auditory cortex (AI). We also show that auditory experience in freely-behaving animals engages estrogen-producing and -sensitive neurons in AI. These estrogen-associated networks are greatly stable, and do not quantitatively change as a result of acute episodes of sensory experience. We further demonstrate the neurochemical identity of estrogen-producing and estrogen-sensitive neurons in AI and show that these cell populations are phenotypically distinct. Our findings provide the first direct demonstration that estrogen-associated circuits are highly prevalent and engaged by sensory experience in the mouse auditory cortex, and suggest that previous correlations between estrogen levels and hearing function may be related to brain-generated hormone production. Finally, our findings suggest that estrogenic modulation may be a central component of the operational framework of central auditory networks. PMID:22545003

Tremere, Liisa A; Burrows, Kaiping; Jeong, Jin-Kwon; Pinaud, Raphael

2011-10-25

227

Organization of Estrogen-Associated Circuits in the Mouse Primary Auditory Cortex  

PubMed Central

Sex steroid hormones influence the perceptual processing of sensory signals in vertebrates. In particular, decades of research have shown that circulating levels of estrogen correlate with hearing function. The mechanisms and sites of action supporting this sensory-neuroendocrine modulation, however, remain unknown. Here we combined a molecular cloning strategy, fluorescence in-situ hybridization and unbiased quantification methods to show that estrogen-producing and -sensitive neurons heavily populate the adult mouse primary auditory cortex (AI). We also show that auditory experience in freely-behaving animals engages estrogen-producing and -sensitive neurons in AI. These estrogen-associated networks are greatly stable, and do not quantitatively change as a result of acute episodes of sensory experience. We further demonstrate the neurochemical identity of estrogen-producing and estrogen-sensitive neurons in AI and show that these cell populations are phenotypically distinct. Our findings provide the first direct demonstration that estrogen-associated circuits are highly prevalent and engaged by sensory experience in the mouse auditory cortex, and suggest that previous correlations between estrogen levels and hearing function may be related to brain-generated hormone production. Finally, our findings suggest that estrogenic modulation may be a central component of the operational framework of central auditory networks.

Tremere, Liisa A.; Burrows, Kaiping; Jeong, Jin-Kwon; Pinaud, Raphael

2011-01-01

228

Immortalized mouse dental papilla mesenchymal cells preserve odontoblastic phenotype and respond to bone morphogenetic protein 2.  

PubMed

Odontogenesis is the result of the reciprocal interactions between epithelial-mesenchymal cells leading to terminally differentiated odontoblasts. This process from dental papilla mesenchymal cells to odontoblasts is regulated by a complex signaling pathway. When isolated from the developing tooth germs, odontoblasts quickly lose their potential to maintain the odontoblast-specific phenotype. Therefore, generation of an odontoblast-like cell line would be a good surrogate model for studying the dental mesenchymal cell differentiation into odontoblasts and the molecular events of dentin formation. In this study, immortalized dental papilla mesenchymal cell lines were generated from the first mouse mandibular molars at postnatal day 3 using pSV40. These transformed cells were characterized by RT-PCR, immunohistochemistry, Western blot, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iMDP-3, displayed a high proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers and demonstrated the ability to differentiate and form mineralized nodules. Furthermore, iMDP-3 cells had high transfection efficiency as well as were inducible and responded to BMP2 stimulation. We conclude that the establishment of the stable murine dental papilla mesenchymal cell line might be used for studying the mechanisms of dental cell differentiation and dentin formation. PMID:23813243

Wang, Feng; Wu, Li-An; Li, Wentong; Yang, Yuan; Guo, Feng; Gao, Qingping; Chuang, Hui-Hsiu; Shoff, Lisa; Wang, Wei; Chen, Shuo

2013-09-01

229

Protein tyrosine phosphatase profiling studies during brown adipogenic differentiation of mouse primary brown preadipocytes.  

PubMed

There is a correlation between obesity and the amount of brown adipose tissue; however, the molecular mechanism of brown adipogenic differentiation has not been as extensively studied. In this study, we performed a protein tyrosine phosphatase (PTP) profiling analysis during the brown adipogenic differentiation of mouse primary brown preadipocytes. Several PTPs, including PTPRF, PTPRZ, and DUSP12 showing differential expression patterns were identified. In the case of DUSP12, the expression level is dramatically downregulated during brown adipogenesis. The ectopic expression of DUSP12 using a retroviral expression system induces the suppression of adipogenic differentiation, whereas a catalytic inactive DUSP12 mutant showed no effect on differentiation. These results suggest that DUSP12 is involved in brown adipogenic differentiation and may be used as a target protein for the treatment or prevention of obesity by the regulation of brown adipogenic differentiation. PMID:24152912

Choi, Hye-Ryung; Kim, Won Kon; Park, Anna; Jung, Hyeyun; Han, Baek Soo; Lee, Sang Chul; Bae, Kwang-Hee

2013-11-01

230

Sensorimotor mismatch signals in primary visual cortex of the behaving mouse.  

PubMed

Studies in anesthetized animals have suggested that activity in early visual cortex is mainly driven by visual input and is well described by a feedforward processing hierarchy. However, evidence from experiments on awake animals has shown that both eye movements and behavioral state can strongly modulate responses of neurons in visual cortex; the functional significance of this modulation, however, remains elusive. Using visual-flow feedback manipulations during locomotion in a virtual reality environment, we found that responses in layer 2/3 of mouse primary visual cortex are strongly driven by locomotion and by mismatch between actual and expected visual feedback. These data suggest that processing in visual cortex may be based on predictive coding strategies that use motor-related and visual input to detect mismatches between predicted and actual visual feedback. PMID:22681686

Keller, Georg B; Bonhoeffer, Tobias; Hübener, Mark

2012-06-01

231

Subpopulations of mouse bone marrow high-proliferative-potential colony-forming cells.  

PubMed

Bone marrow cells taken from mice treated eight days previously with 5-fluorouracil, formed colonies consisting of 10-100 cells after four days of incubation in methylcellulose cultures containing only 500 cells/dish, in the presence of partially purified synergistic factor from human placental-conditioned medium (SFHPlac) and macrophage colony-stimulating factor (CSF-1). Replating of these colonies revealed a high incidence (27%) of another class of high-proliferative-potential colony-forming cells (HPP-CFC) responsive only to the synergistic factor in WEHI-3B-conditioned medium (SFW, which appears to be identical to interleukin 3) plus CSF-1. These colonies contained no HPP-CFC responsive to SFHPlac plus CSF-1, although primary cultures incubated for 14 days in the presence of SFHPlac plus CSF-1 formed large colonies (diameter greater than 0.5 mm), indicating the presence of HPP-CFC responsive to SFHPlac plus CSF-1 in the starting marrow. Primary cultures containing SFW alone, or purified interleukin 3 alone, also gave rise to colonies consisting of 10-100 cells after four days of incubation in methylcellulose cultures; however, the cells from these colonies were unable to form large colonies on replating in the presence of either CSF-1 plus SFHPlac or CSF-1 plus SFW. These results suggest that two distinct populations of HPP-CFC exist and that the population of HPP-CFC stimulated by CSF-1 plus SFHPlac differentiates to form HPP-CFC that respond to CSF-1 plus SFW. PMID:3489636

McNiece, I K; Bradley, T R; Kriegler, A B; Hodgson, G S

1986-10-01

232

Modeling Pathogenesis of Primary Liver Cancer in Lineage-Specific Mouse Cell Types  

PubMed Central

BACKGROUND & AIMS Human primary liver cancer (PLC) is classified into biologically distinct subgroups, based on cellular origin. Liver cancer stem cells (CSCs) have been recently described. We investigated the ability of distinct lineages of hepatic cells to become liver CSCs and the phenotypic and genetic heterogeneity of PLC. METHODS We transduced mouse primary hepatic progenitor cells (HPC), lineage-committed hepatoblasts, and differentiated adult hepatocytes with transgenes encoding oncogenic H-Ras and simian virus 40 large-T antigen. The CSC properties of transduced cells and their ability to form tumors were tested by standard in vitro and in vivo assays and transcriptome profiling. RESULTS Irrespective of origin, all transduced cells acquired markers of CSC/progenitor cells, side populations, and self-renewal capacity in vitro. They also formed a broad spectrum of liver tumors, ranging from cholangiocarcinoma to hepatocellular carcinoma, which resembled human liver tumors, based on genomic and histologic analyses. The tumor cells co-expressed hepatocyte (HNF4A), biliary progenitor cell (keratin 19, EpCAM, A6), and mesenchyme (vimentin) markers and showed disregulation of genes that control the epithelial–mesenchymal transition. Gene expression analyses could distinguish tumors of different cellular origin, indicating the contribution of lineage-stage dependent genetic changes to malignant transformation. Activation of c-Myc and its target genes was required to reprogram adult hepatocytes into CSC and for tumors to develop. Stable knockdown of c-Myc in transformed adult hepatocytes reduced their CSC properties in vitro and suppressed growth of tumors in immunodeficient mice. CONCLUSIONS Any cell type in the mouse hepatic lineage can undergo oncogenic reprogramming into a CSC, by activating different cell type-specific pathways. Identification of common and cell-of-origin specific phenotypic and genetic changes could provide new therapeutic targets for liver cancer.

Holczbauer, Agnes; Factor, Valentina M.; Andersen, Jesper B.; Marquardt, Jens U.; Kleiner, David; Raggi, Chiara; Kitade, Mitsuteru; Seo, Daekwan; Akita, Hirofumi; Durkin, Marian; Thorgeirsson, Snorri S.

2013-01-01

233

A humanized mouse identifies the bone marrow as a niche with low therapeutic IgG activity.  

PubMed

Genetic differences between humans and in vivo model systems, including mice and nonhuman primates, make it difficult to predict the efficacy of immunoglobulin G (IgG) activity in humans and understand the molecular and cellular mechanisms underlying that activity. To bridge this gap, we established a small-animal model system that allowed us to study human IgG effector functions in the context of an intact human immune system without the interference of murine Fc? receptors expressed on mouse innate immune effector cells in vivo. Using a model of B cell depletion with different human IgG variants that recognize CD20, we show that this humanized mouse model can provide unique insights into the mechanism of human IgG activity in vivo. Importantly, these studies identify the bone marrow as a niche with low therapeutic IgG activity. PMID:24685130

Lux, Anja; Seeling, Michaela; Baerenwaldt, Anne; Lehmann, Birgit; Schwab, Inessa; Repp, Roland; Meidenbauer, Norbert; Mackensen, Andreas; Hartmann, Arndt; Heidkamp, Gordon; Dudziak, Diana; Nimmerjahn, Falk

2014-04-10

234

Primary Epiphyseal Arteriopathy in a Mouse Model of Steroid-Induced Osteonecrosis  

PubMed Central

Patients undergoing glucocorticoid therapy for a variety of disorders, including autoimmune diseases and hematological malignancies, are at risk of developing osteonecrosis. Despite extensive research in both patients and animal models, the underlying pathogenesis remains unclear. Proposed inciting mechanisms include intravascular thrombotic occlusion, marrow fat hypertrophy, osteocyte and/or endothelial cell apoptosis, hypercoagulability, and vasoconstriction of specific arteries and arterioles supplying bone. Our laboratory has developed a model of steroid-induced osteonecrosis in BALBcJ mice which reflects clinically relevant exposures to glucocorticoids in which treated mice develop osteonecrosis of the distal femoral epiphysis when administered 4 to 8 mg/L dexamethasone in drinking water for 6 weeks. We identified lesions in arterioles supplying this area, with the mildest occurring in knees without any evidence of osteonecrosis. However, arteriopathy was more common among mice that did versus did not develop osteonecrosis (P < 0.0001); in mice with osteonecrosis, the associated vessels showed transmural necrosis and thickening of the vessel wall progressing to the point of luminal obstruction. In the most severe cases of osteonecrosis, end-stage lesions consisted of fully occluded vessels with marrow and bone necrosis involving the entire epiphysis. We propose that a primary arteriopathy is the initiating event in the genesis of steroid-induced osteonecrosis and provides a basis for future investigation of this disease process.

Janke, Laura J.; Liu, Chengcheng; Vogel, Peter; Kawedia, Jitesh; Boyd, Kelli L.; Funk, Amy J.; Relling, Mary V.

2014-01-01

235

Establishment of a mast cell line, NCL-2, without Kit mutation, derived from NC mouse bone marrow  

PubMed Central

Immortal mast cell lines, such as RBL-2H3 and HMC-1 cells, are commonly utilized to investigate the function of mast cells. However, they are tumor cells carrying a gain-of-function mutation of Kit. We established an immortal mast cell line without Kit mutation, NCL-2, derived from NC mouse bone marrow. NCL-2 cells could be maintained without additional growth factors and thus could respond to exogenous growth signals. Moreover, NCL-2 cells expressed Fc?RI and KIT, and release histamine and LTB4 in response to antigen stimulation. This cell line could be a useful tool to analyze proliferation, differentiation, and function of normal mast cells.

Hiragun, Takaaki; Yanase, Yuhki; Okabe, Tsutomu; Hiragun, Makiko; Kawai, Mikio; Hide, Michihiro

2014-01-01

236

Bone regeneration of mouse critical-sized calvarial defects with human mesenchymal stem cells in scaffold  

PubMed Central

Combination of tissue engineering and cell therapy represents a promising approach for bone regeneration. Human mesenchymal stem cells (hMSCs) have properties that include low immunogenicity, high proliferation rate, and multi-differentiation potential; therefore, they are an attractive seeding source for tissue engineering therapy. Here we found that hMSCs with a scaffold did not affect cell viability and osteogenic differentiation. We also investigated regenerative effect of hMSCs with the scaffold in a calvarial bone defect model. Formation of new bone was evaluated by micro-CT, histology and expression of osteogenic markers. The results clearly showed interesting evidence indicating that hMSCs with scaffold increased the formation of new bone and expression of osteogenic markers, compared to the empty and scaffold only groups. Overall, our results suggest that hMSCs with scaffold are suitable for stimulation of intense bone regeneration in critical-sized bone defects.

Im, Jin-Young; Min, Woo-Kie; You, Changkook; Kim, Hyun-Ok; Jin, Hee-Kyung

2013-01-01

237

Isolation and characterization of resident stromal macrophages and hematopoietic cell clusters from mouse bone marrow  

PubMed Central

In situ studies with the mouse macrophage (M phi)-specific antibody, F4/80, have shown that resident M phi in femoral bone marrow (RBMM) form hematopoietic islands with immature myelomonocytic and erythroid cells (Hume, D. A., et al. 1983. J. Exp. Med. 158: 1522). We have isolated these islands (clusters) by collagenase digestion, purified them from single cells by velocity sedimentation, and analyzed their cellular content. The clusters, ranging from 5- to 100 cells, constituted approximately 7% of the total nucleated cells, and greater than 70% contained at least one strongly staining, F4/80+ central M phi. In comparison, less than 26% showed reactivity for alkaline phosphatase, a marker of fibroblastoid reticulum cells. Compared with the nonclustering population, clusters were enriched with RBMM, fibroblastoid cells, and immature hematopoietic cells, but depleted of mature granulocytes and erythrocytes. The RBMM population was purified from other cells in clusters by selective adherence to glass and was compared with resident peritoneal M phi (RPM) for morphology and the presence of antigens, receptors, and enzymes. RBMM spread more extensively than RPM and frequently extended delicate plasma membrane processes. These and subsequent differences were not attributable to the collagenase treatment. Both M phi populations stained positively with antibodies F4/80 and 2.4G2 (Fc receptor IgG1/2b), bore mannosyl/fucosyl receptors, and showed reactivity for acid phosphatase and nonspecific esterase I. In contrast to RPM, RBMM had no detectable Mac-1 antigen (CR3) or complement receptors, but bore higher levels of Fc receptors (IgG2a and IgG2b) and Ia antigens. In addition, RBMM possessed a novel hemagglutinin activity for unopsonized sheep erythrocytes, which was not present on RPM. RBMM showed no respiratory burst activity in response to zymosan particles, but ingested them avidly. The growth properties of clustering and nonclustered populations were compared by measurement of [3H]thymidine incorporation and progenitor assays. Cells in clusters incorporated three- to fourfold more thymidine than nonclustered cells even in the absence of exogenous growth factors, and autoradiography demonstrated that RBMM made contact with proliferating cells. In contrast, the clusters contained over threefold fewer granulocyte/M phi progenitors compared with nonclustering cells. When clusters were cultivated for up to 3 d, there was rapid outgrowth of monocytes and fibroblastoid cells. These studies demonstrate that RBMM bear a distinct morphology and phenotype.(ABSTRACT TRUNCATED AT 400 WORDS)

1985-01-01

238

MOUSE  

NSDL National Science Digital Library

Based in New York City, the MOUSE organization works to empower "underserved students to provide technology support and leadership in their schools, supporting their academic and career success." On their homepage, visitors can learn about their programs, learn about supporting the MOUSE organization, and read up on their resources. In the "Resources" area, visitors can learn about their outreach activities in New York City, Chicago, and California. Visitors working in educational outreach will appreciate the information offered here, including materials on how different groups can receive assistance from the MOUSE organization. Also, visitors can look over the "News" updates to learn about their new programs, their educational seminars, and their outreach activities.

239

Therapeutic impact of low amplitude high frequency whole body vibrations on the osteogenesis imperfecta mouse bone?  

PubMed Central

Osteogenesis imperfecta (OI) is characterized by extremely brittle bone. Currently, bisphosphonate drugs allow a decrease of fracture by inhibiting bone resorption and increasing bone mass but with possible long term side effects. Whole body mechanical vibrations (WBV) treatment may offer a promising route to stimulate bone formation in OI patients as it has exhibited health benefits on both muscle and bone mass in human and animal models. The present study has investigated the effects of WBV (45 Hz, 0.3 g, 15 minutes/days, 5 days/week) in young OI (oim) and wild type female mice from 3 to 8 weeks of age. Vibration therapy resulted in a significant increase in the cortical bone area and cortical thickness in the femur and tibia diaphysis of both vibrated oim and wild type mice compared to sham controls. Trabecular bone was not affected by vibration in the wild type mice; vibrated oim mice, however, exhibited significantly higher trabecular bone volume fraction in the proximal tibia. Femoral stiffness and yield load in three point bending were greater in the vibrated wild type mice than in sham controls, most likely attributed to the increase in femur cortical cross sectional area observed in the ?CT morphology analyses. The vibrated oim mice showed a trend toward improved mechanical properties, but bending data had large standard deviations and there was no significant difference between vibrated and non-vibrated oim mice. No significant difference of the bone apposition was observed in the tibial metaphyseal trabecular bone for both the oim and wild type vibrated mice by histomorphometry analyses of calcein labels. At the mid diaphysis, the cortical bone apposition was not significantly influenced by the WBV treatment in both the endosteum and periosteum of the oim vibrated mice while a significant change is observed in the endosteum of the vibrated wild type mice. As only a weak impact in bone apposition between the vibrated and sham groups is observed in the histological sections, it is possible that WBV reduced bone resorption, resulting in a relative increase in cortical thickness. Whole body vibration appears as a potential effective and innocuous means for increasing bone formation and strength, which is particularly attractive for treating the growing skeleton of children suffering from brittle bone disease or low bone density pathologies without the long term disadvantages of current pharmacological therapies.

Vanleene, Maximilien; Shefelbine, Sandra J.

2013-01-01

240

Therapeutic impact of low amplitude high frequency whole body vibrations on the osteogenesis imperfecta mouse bone.  

PubMed

Osteogenesis imperfecta (OI) is characterized by extremely brittle bone. Currently, bisphosphonate drugs allow a decrease of fracture by inhibiting bone resorption and increasing bone mass but with possible long term side effects. Whole body mechanical vibrations (WBV) treatment may offer a promising route to stimulate bone formation in OI patients as it has exhibited health benefits on both muscle and bone mass in human and animal models. The present study has investigated the effects of WBV (45Hz, 0.3g, 15minutes/days, 5days/week) in young OI (oim) and wild type female mice from 3 to 8weeks of age. Vibration therapy resulted in a significant increase in the cortical bone area and cortical thickness in the femur and tibia diaphysis of both vibrated oim and wild type mice compared to sham controls. Trabecular bone was not affected by vibration in the wild type mice; vibrated oim mice, however, exhibited significantly higher trabecular bone volume fraction in the proximal tibia. Femoral stiffness and yield load in three point bending were greater in the vibrated wild type mice than in sham controls, most likely attributed to the increase in femur cortical cross sectional area observed in the ?CT morphology analyses. The vibrated oim mice showed a trend toward improved mechanical properties, but bending data had large standard deviations and there was no significant difference between vibrated and non-vibrated oim mice. No significant difference of the bone apposition was observed in the tibial metaphyseal trabecular bone for both the oim and wild type vibrated mice by histomorphometry analyses of calcein labels. At the mid diaphysis, the cortical bone apposition was not significantly influenced by the WBV treatment in both the endosteum and periosteum of the oim vibrated mice while a significant change is observed in the endosteum of the vibrated wild type mice. As only a weak impact in bone apposition between the vibrated and sham groups is observed in the histological sections, it is possible that WBV reduced bone resorption, resulting in a relative increase in cortical thickness. Whole body vibration appears as a potential effective and innocuous means for increasing bone formation and strength, which is particularly attractive for treating the growing skeleton of children suffering from brittle bone disease or low bone density pathologies without the long term disadvantages of current pharmacological therapies. PMID:23352925

Vanleene, Maximilien; Shefelbine, Sandra J

2013-04-01

241

Phenotypic Correction of a Mouse Model for Primary Hyperoxaluria With Adeno-associated Virus Gene Transfer  

PubMed Central

Primary hyperoxaluria type I (PH1) is an inborn error of metabolism caused by deficiency of the hepatic enzyme alanine-glyoxylate aminotransferase (AGXT or AGT) which leads to overproduction of oxalate by the liver and subsequent urolithiasis and renal failure. The current therapy largely depends on liver transplantation, which is associated with significant morbidity and mortality. To explore an alternative treatment, we used somatic gene transfer in a mouse genetic model for PH1 (Agxt1KO). Recombinant adeno-associated virus (AAV) vectors containing the human AGXT complementary DNA (cDNA) were pseudotyped with capsids from either serotype 8 or 5, and delivered to the livers of Agxt1KO mice via the tail vein. Both AAV8-AGXT and AAV5-AGXT vectors were able to reduce oxaluria to normal levels. In addition, treated mice showed blunted increase of oxaluria after challenge with ethylene glycol (EG), a glyoxylate precursor. In mice, AGT enzyme activity in whole liver extracts were restored to normal without hepatic toxicity nor immunogenicity for the 50 day follow-up. In summary, this study demonstrates the correction of primary hyperoxaluria in mice treated with either AAV5 or AAV8 vectors.

Salido, Eduardo; Rodriguez-Pena, Marisol; Santana, Alfredo; Beattie, Stuart G.; Petry, Harald; Torres, Armando

2011-01-01

242

Evaluation of the correlation between insertion torque and primary stability of dental implants using a block bone test  

PubMed Central

Purpose Implant stability at the time of surgery is crucial for the long-term success of dental implants. Primary stability is considered of paramount importance to achieve osseointegration. The purpose of the present study was to investigate the correlation between the insertion torque and primary stability of dental implants using artificial bone blocks with different bone densities and compositions to mimic different circumstances that are encountered in routine daily clinical settings. Methods In order to validate the objectives, various sized holes were made in bone blocks with different bone densities (#10, #20, #30, #40, and #50) using a surgical drill and insertion torque together with implant stability quotient (ISQ) values that were measured using the Osstell Mentor. The experimental groups under evaluation were subdivided into 5 subgroups according to the circumstances. Results In group 1, the mean insertion torque and ISQ values increased as the density of the bone blocks increased. For group 2, the mean insertion torque values decreased as the final drill size expanded, but this was not the case for the ISQ values. The mean insertion torque values in group 3 increased with the thickness of the cortical bone, and the same was true for the ISQ values. For group 4, the mean insertion torque values increased as the cancellous bone density increased, but the correlation with the ISQ values was weak. Finally, in group 5, the mean insertion torque decreased as the final drill size increased, but the correlation with the ISQ value was weak. Conclusions Within the limitations of the study, it was concluded that primary stability does not simply depend on the insertion torque, but also on the bone quality.

Bayarchimeg, Dorjpalam; Namgoong, Hee; Kim, Byung Kook; Kim, Myung Duk; Kim, Sungtae; Kim, Tae-Il; Seol, Yang Jo; Lee, Yong Moo; Ku, Young; Rhyu, In-Chul; Lee, Eun Hee

2013-01-01

243

Disrupted bone remodeling leads to cochlear overgrowth and hearing loss in a mouse model of fibrous dysplasia.  

PubMed

Normal hearing requires exquisite cooperation between bony and sensorineural structures within the cochlea. For example, the inner ear secretes proteins such as osteoprotegrin (OPG) that can prevent cochlear bone remodeling. Accordingly, diseases that affect bone regulation can also result in hearing loss. Patients with fibrous dysplasia develop trabecular bone overgrowth resulting in hearing loss if the lesions affect the temporal bones. Unfortunately, the mechanisms responsible for this hearing loss, which could be sensorineural and/or conductive, remain unclear. In this study, we used a unique transgenic mouse model of increased Gs G-protein coupled receptor (GPCR) signaling induced by expression of an engineered receptor, Rs1, in osteoblastic cells. These ColI(2.3)+/Rs1+ mice showed dramatic bone lesions that histologically and radiologically resembled fibrous dysplasia. We found that ColI(2.3)+/Rs1+ mice showed progressive and severe conductive hearing loss. Ossicular chain impingement increased with the size and number of dysplastic lesions. While sensorineural structures were unaffected, ColI(2.3)+/Rs1+ cochleae had abnormally high osteoclast activity, together with elevated tartrate resistant acid phosphatase (TRAP) activity and receptor activator of nuclear factor kappa-B ligand (Rankl) mRNA expression. ColI(2.3)+/Rs1+ cochleae also showed decreased expression of Sclerostin (Sost), an antagonist of the Wnt signaling pathway that normally increases bone formation. The osteocyte canalicular networks of ColI(2.3)+/Rs1+ cochleae were disrupted and showed abnormal osteocyte morphology. The osteocytes in the ColI(2.3)+/Rs1+ cochleae showed increased expression of matrix metalloproteinase 13 (MMP-13) and TRAP, both of which can support osteocyte-mediated peri-lacunar remodeling. Thus, while the ossicular chain impingement is sufficient to account for the progressive hearing loss in fibrous dysplasia, the deregulation of bone remodeling extends to the cochlea as well. Our findings suggest that factors regulating bone remodeling, including peri-lacunar remodeling by osteocytes, may be useful targets for treating the bony overgrowths and hearing changes of fibrous dysplasia and other bony pathologies. PMID:24788917

Akil, Omar; Hall-Glenn, Faith; Chang, Jolie; Li, Alfred; Chang, Wenhan; Lustig, Lawrence R; Alliston, Tamara; Hsiao, Edward C

2014-01-01

244

Disrupted Bone Remodeling Leads to Cochlear Overgrowth and Hearing Loss in a Mouse Model of Fibrous Dysplasia  

PubMed Central

Normal hearing requires exquisite cooperation between bony and sensorineural structures within the cochlea. For example, the inner ear secretes proteins such as osteoprotegrin (OPG) that can prevent cochlear bone remodeling. Accordingly, diseases that affect bone regulation can also result in hearing loss. Patients with fibrous dysplasia develop trabecular bone overgrowth resulting in hearing loss if the lesions affect the temporal bones. Unfortunately, the mechanisms responsible for this hearing loss, which could be sensorineural and/or conductive, remain unclear. In this study, we used a unique transgenic mouse model of increased Gs G-protein coupled receptor (GPCR) signaling induced by expression of an engineered receptor, Rs1, in osteoblastic cells. These ColI(2.3)+/Rs1+ mice showed dramatic bone lesions that histologically and radiologically resembled fibrous dysplasia. We found that ColI(2.3)+/Rs1+ mice showed progressive and severe conductive hearing loss. Ossicular chain impingement increased with the size and number of dysplastic lesions. While sensorineural structures were unaffected, ColI(2.3)+/Rs1+ cochleae had abnormally high osteoclast activity, together with elevated tartrate resistant acid phosphatase (TRAP) activity and receptor activator of nuclear factor kappa-B ligand (Rankl) mRNA expression. ColI(2.3)+/Rs1+ cochleae also showed decreased expression of Sclerostin (Sost), an antagonist of the Wnt signaling pathway that normally increases bone formation. The osteocyte canalicular networks of ColI(2.3)+/Rs1+ cochleae were disrupted and showed abnormal osteocyte morphology. The osteocytes in the ColI(2.3)+/Rs1+ cochleae showed increased expression of matrix metalloproteinase 13 (MMP-13) and TRAP, both of which can support osteocyte-mediated peri-lacunar remodeling. Thus, while the ossicular chain impingement is sufficient to account for the progressive hearing loss in fibrous dysplasia, the deregulation of bone remodeling extends to the cochlea as well. Our findings suggest that factors regulating bone remodeling, including peri-lacunar remodeling by osteocytes, may be useful targets for treating the bony overgrowths and hearing changes of fibrous dysplasia and other bony pathologies.

Chang, Jolie; Li, Alfred; Chang, Wenhan; Lustig, Lawrence R.; Alliston, Tamara; Hsiao, Edward C.

2014-01-01

245

Intrinsic differences in BRITE adipogenesis of primary adipocytes from two different mouse strains.  

PubMed

BRITE (brown-in-white) cells are brown adipocyte-like cells found in white adipose tissue (WAT) of rodents and/or humans. The recruitment of BRITE adipocytes, referred to as the browning of WAT, is hallmarked by the expression of UCP1 and exerts beneficial metabolic effects. Here we address whether beyond systemic cues depot- and strain-specific variation in BRITE recruitment is determined by a cellular program intrinsic to progenitors. Therefore we compared the browning capacity of serum and investigated brown and BRITE adipogenesis in primary cultures of stromal-vascular cells isolated from interscapular brown adipose tissue (iBAT), inguinal white adipose tissue (iWAT) and epididymal white adipose tissue (eWAT) in two inbred mouse strains C57BL/6J (B6, a strain with low browning propensity) and 129/S6SvEv (129, a strain with high browning propensity). Paradoxically, serum collected from B6 mice was more potent in the promotion of browning than serum collected from 129 mice. Nevertheless, we demonstrate that depot- and strain-specific differences observed in vivo are pheno-copied in primary cultures in vitro, as judged by UCP1 expression and by functional analysis. Notably, primary adipocytes from 129 mice had a higher capacity for isoproterenol-induced uncoupled respiration than B6. We conclude that cues intrinsic to the progenitor cells contribute to differential BRITE adipogenesis. Further analyses demonstrate that these cues are independent of autocrine/paracrine mechanisms, BRITE progenitor abundance and genetic variation in the gene regulatory region of Ucp1 but rather depend on trans-acting factors. These results provide new insights on the molecular basis of strain and depot-specific differences in BRITE adipogenesis. PMID:24953778

Li, Yongguo; Bolze, Florian; Fromme, Tobias; Klingenspor, Martin

2014-09-01

246

Upregulating CXCR4 in Human Fetal Mesenchymal Stem Cells Enhances Engraftment and Bone Mechanics in a Mouse Model of Osteogenesis Imperfecta  

PubMed Central

Stem cells have considerable potential to repair damaged organs and tissues. We previously showed that prenatal transplantation of human first trimester fetal blood mesenchymal stem cells (hfMSCs) in a mouse model of osteogenesis imperfecta (oim mice) led to a phenotypic improvement, with a marked decrease in fracture rate. Donor cells differentiated into mature osteoblasts, producing bone proteins and minerals, including collagen type I?2, which is absent in nontransplanted mice. This led to modifications of the bone matrix and subsequent decrease of bone brittleness, indicating that grafted cells directly contribute to improvement of bone mechanical properties. Nevertheless, the therapeutic effect was incomplete, attributing to the limited level of engraftment in bone. In this study, we show that although migration of hfMSCs to bone and bone marrow is CXCR4-SDF1 (SDF1 is stromal-derived factor) dependent, only a small number of cells present CXCR4 on the cell surface despite high levels of internal CXCR4. Priming with SDF1, however, upregulates CXCR4 to increase the CXCR4+ cell fraction, improving chemotaxis in vitro and enhancing engraftment in vivo at least threefold in both oim and wild-type bone and bone marrow. Higher engraftment in oim bones was associated with decreased bone brittleness. This strategy represents a step to improve the therapeutic benefits of fetal cell therapy toward being curative.

Jones, Gemma N.; Moschidou, Dafni; Lay, Kenneth; Abdulrazzak, Hassan; Vanleene, Maximilien; Shefelbine, Sandra J.; Polak, Julia; de Coppi, Paolo; Fisk, Nicholas M.

2012-01-01

247

Prophylactic use of antibiotic-loaded bone cement in primary total knee arthroplasty: Justified or not?  

PubMed Central

Background: The routine use of antibiotic-loaded bone cement (ABLC) during primary or uninfected revision arthroplasty remains controversial. Many studies quote the total joint arthroplasty (TJA) infection rate to be less than 1%. Total knee arthroplasty (TKA) has a higher infection rate than total hip arthroplasty (THA). Based on both animal and human studies in the past, ABLC has been found effective in reducing the risk of infection in primary TJA. We are presenting retrospective analysis of results in terms of infection rate in 659 TKA performed by a single surgeon under similar conditions during 2004–2007 using CMW1 (Depuy, Leeds, UK) with premixed 1 g of gentamicin. Patients and Methods: We did primary TKA in 659 knees of 379 patients during 2004–2007 using CMW1 (Depuy, Leeds, UK) cement containing 1 g of gentamicin in 40 g of cement in a premixed form. Standard OT conditions were maintained using laminar air flow, isolation suits for the operating team, pulse lavage and disposable drapes in each patients. Midvastus approach was used in all the patients to expose the knee joint. A systemic antibiotic (third-generation cephalosporin and aminoglycoside) was used preoperatively and 48 h postoperatively. We observed the patients in terms of infection in the high-risk and low-risk group till the recent follow-up with a mean of 20.6 months (9–38 months). Results: We had deep infection in six knees in six patients and all of them required two-stage revision surgery later in the high-risk group. Infection occurred at a mean of 20.5 months after surgery earliest at 9 months and latest at 36 months after surgery. The infection rate in our study was 0.91% which is comparatively less than the reported incidence of 1–2% in reported studies. Conclusion: We conclude that the use of antibiotic loaded bone cement is one of the effective means in preventing infection in primary TJA.

Srivastav, Amit K; Nadkarni, Biren; Srivastav, Shekhar; Mittal, Vivek; Agarwal, Shekhar

2009-01-01

248

Generation of mouse and human induced pluripotent stem cells (iPSC) from primary somatic cells.  

PubMed

Cellular reprogramming consists of the conversion of differentiated cells into pluripotent cells; the so-called induced Pluripotent Stem Cells. iPSC are amenable to in vitro manipulation and, in theory, direct production of any differentiated cell type. Furthermore, iPSC can be obtained from sick individuals and subsequently used for disease modeling, drug discovery and regenerative treatments. iPSC production was first achieved by transducing, with the use of retroviral vectors, four specific transcription factors: Oct4, Klf4, Sox2 and c-Myc (OKSM), into primary cells in culture Takahashi and Yamanaka, (Cell 126(4):663-676, 2006). Many alternative protocols have since been proposed: repeated transfections of expression plasmids containing the four pluripotency-associated genes Okita et al. (Science 322(5903):949-953, 2008), lentiviral delivery of the four factors Sommer et al. (Stem Cells 27(3):543-549, 2009), Sendai virus delivery Fusaki et al. (Proceedings of the Japan Academy. Series B, Physical and Biological Sciences 85(8):348-362, 2009), removal of the reprogramming vectors by 'piggyBac' transposition Woltjen et al. (Nature 458(7239):766-770, 2009); Kaji et al. (Nature 458(7239):771-775, 2009), Cre-recombinase excisable viruses Soldner et al. (Cell 136(5):964-977, 2009), episomal vectors Yu et al. (Science 324(5928):797-801, 2009), cell-penetrating reprogramming proteins Zhou et al. (Stem Cells 4(5):381-384, 2009), mammalian artificial chromosomes Hiratsuka et al. (PLoS One 6(10):e25961, 2011) synthetically modified mRNAs Warren et al. (Scientific Reports 2:657, 2012), miRNA Anokye-Danso et al. (Cell Stem Cell 8(4):376-388, 2009); however, although some of these methods are commercially available, in general they still need to attain the reproducibility and reprogramming efficiency required for routine applications Mochiduki and Okita (Biotechnol Journal 7(6):789-797, 2012). Herein we explain, in four detailed protocols, the isolation of mouse and human somatic cells and their reprogramming into iPSC. All-encompassing instructions, not previously published in a single document, are provided for mouse and human iPSC colony isolation and derivation. Although mouse and human iPSC share similarities in the cellular reprogramming process and culture, both cell types need to be handled differently. PMID:23104133

Lorenzo, I M; Fleischer, A; Bachiller, D

2013-08-01

249

GlaI digestion of mouse ?-satellite DNA: study of primary structure and ACGT sites methylation  

Microsoft Academic Search

BACKGROUND: Patterns of mouse DNA hydrolysis with restriction enzymes are coincided with calculated diagrams of genomic DNA digestion in silico, except presence of additional bright bands, which correspond to monomer and dimer of ?-satellite DNA. Only small portion of mouse ?-satellite DNA sequences are presented in databases. Methyl-directed endonuclease GlaI cleaves mouse DNA and may be useful for a detailed

Murat A Abdurashitov; Valery A Chernukhin; Danila A Gonchar; Sergey Kh Degtyarev

2009-01-01

250

The monoclonal antibodies 18d7/91f2 recognize a receptor regulatory protein on mouse bone marrow stromal cells.  

PubMed

Two monoclonal antibodies 18D7 and 91F2 were developed by immunizing rats with the mouse bone marrow-derived osteogenic cell line MN7. Hybridomas secreting rat antibodies against MN7 cell surface markers were selected by flow cytometry analysis. Both the monoclonal antibody 18D7 and the monoclonal antibody 91F2 are directed against the same cell surface antigen present on MN7 cells. Here, we report on the immunopurification of the 18D7/91F2 antigen and its identification as the prostaglandin F2 alpha receptor regulatory protein (FPRP). FPRP is expressed as a single messenger RNA (mRNA) of approximately 6 kilobases (kb) in MN7 cells and is differentially expressed in developing osteogenic cultures of bone marrow cells of the mouse. However, addition of the monoclonal antibodies 18D7 and 91F2 to these cultures did not inhibit bone formation in vitro. Both monoclonal antibodies reacted with mouse stromal cell lines established from bone marrow, thymus, spleen, and mandibular condyles. Immunohistochemical analysis of mature tibia of mice using the monoclonal antibody 18D7 revealed the presence of a distinct population of bone marrow cells close to trabecular and endosteal bone surfaces. In the central bone marrow, hardly any positive cells were found. In 17-day-old fetal mouse radius 18D7 immunoreactivity was restricted to cells in the periosteum in close vicinity to the bone collar. Mature osteoblasts, osteoclasts, osteocytes, growth plate chondrocytes, and mature macrophages were all negative. Taken together, these results suggest that FPRP plays a role in the osteogenic differentiation process. PMID:10893677

Weng, L; Falla, N; Van den Heuvel, R; Raymackers, J; Karperien, M; Van Bezooijen, R; Van Vlasselaer, P; Löwik, C; Merregaert, J

2000-07-01

251

Follow-up of bone lesions in an experimental multiple myeloma mouse model: description of an in vivo technique using radiography dedicated for mammography.  

PubMed Central

The evolution of bone lesions in transplantable C57BL/KaLwRjj 5T mouse myeloma (MM) has been followed in vivo. Mice were anaesthetised and a radiograph of the pelvis and hind legs was performed by a radiograph dedicated for mammography. This is the first description of an in vivo technique under experimental conditions whereby the development of bone lesions owing to the MM growth was demonstrated. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 6

Vanderkerken, K.; Goes, E.; De Raeve, H.; Radl, J.; Van Camp, B.

1996-01-01

252

Cytotoxic effects of propiconazole and its metabolites in mouse and human hepatoma cells and primary mouse hepatocytes  

Microsoft Academic Search

Propiconazole is a triazole-containing fungicide that is used agriculturally on grasses, fruits, grains, seeds, hardwoods, and conifers. Propiconazole is a mouse liver hepatotoxicant and a hepatocarcinogen that has adverse reproductive and developmental toxicities in experimental animals. The goal of this study was to investigate the cytotoxic responses of propiconazole and its metabolites to determine if metabolism of this agent differentially

Pei-Jen Chen; Tanya Moore; Stephen Nesnow

2008-01-01

253

Prognostic significance of soft tissue extension, International Prognostic Index, and multifocality in primary bone lymphoma: a single institutional experience.  

PubMed

Primary bone lymphoma (PBL) is a rare disease. The literature is inconsistent in regard to definition, stage and prognostic factors. We examined the PBL cases seen at the Moffitt Cancer Center between 1998 and 2013 using the 2013 World Health Organization criteria for bone/soft tissue tumours. Seventy PBL patients were included, of whom 53 (75·7%) patients were histologically classified as primary bone diffuse large B-cell lymphoma (PB-DLBCL). Femur was the most commonly involved site in PBLs with unifocal bone lesions, whereas PBLs with multifocal bone lesions most frequently presented with spine disease. Further analysis of the PB-DLBCL subgroup showed that these patients had 3- and 5-year progression-free survival (PFS) of 61·2% and 46·9%, respectively and 5- and 10-year overall survival (OS) of 81·1% and 74·7%, respectively. Multivariate analysis identified soft tissue extension and International Prognostic Index (IPI) score as the most important unfavourable prognostic factors for both PFS and OS. Multifocality was also highly significantly associated with a worse PFS (P = 0·002) and OS (P < 0·001), although it was not identified in multivariate analysis due to its incorporation into the IPI. The results warrant further investigation regarding whether PBL with multifocal bone lesions could be considered as a systemic and more aggressive disease rather than a conventional PBL. PMID:24673481

Wu, Huanwen; Zhang, Ling; Shao, Haipeng; Sokol, Lubomir; Sotomayor, Eduardo; Letson, Douglas; Bui, Marilyn M

2014-07-01

254

Salvage of a femoral nonunion after primary non-Hodgkin's lymphoma of bone: A case report and literature review  

PubMed Central

Summary Background With the advent of superb microsurgery techniques and advanced stabilization instruments, recent decades have seen great progress in treating nonunions secondary to traumatic fractures. However, those nonunions that are secondary to primary non-Hodgkin’s lymphoma of bone and often related to irradiation still remain a challenging problem. The condition could be more perplexing when bone healing abilities are greatly compromised and reliable stabilization is difficult. Case Report We performed an operation using free vascularized fibular graft in combination with a locking plate on a 47-year-old female patient who had suffered from a three-year femoral nonunion after courses of radiochemotherapy for the treatment of primary non-Hodgkin’s lymphoma of bone, a spontaneous femoral shaft fracture, an intramedullary nailing, and some nonoperative interventions in sequence. Primary union of the graft was obtained at 9 months without wound infection. No recurrence of lymphoma occurred in the 61-month follow-up, nor did a stress fracture or failure of fixation. Limb salvage was achieved and the range of motion of the adjacent joints was acceptable. Conclusions Free vascularized fibular graft in combination with a locking plate can effectively enhance bone union in compromised bone and soft tissue milieu. More cases have yet to be further investigated.

Xie, Xue Tao; Gao, You Shui; Zhang, Chang Qing

2011-01-01

255

Array-based comparative genomic hybridisation analysis reveals recurrent chromosomal alterations in primary diffuse large B cell lymphoma of bone  

Microsoft Academic Search

AimsPrimary non-Hodgkin's lymphoma of bone (PLB) is a rare subtype of primary extranodal diffuse large B cell lymphoma. PLB has morphological homogeneity and a relatively favourable clinical behaviour. Recent studies report that array-based comparative genomic hybridisation (array-CGH) analysis can be used to classify lymphomas into clinically and biologically relevant phenotypes and possibly reveal differences in oncogenic mechanisms. Here the authors

F H Heyning; P M Jansen; P C W Hogendoorn; K Szuhai

2010-01-01

256

CCR1 blockade reduces tumor burden and osteolysis in vivo in a mouse model of myeloma bone disease  

PubMed Central

The chemokine CCL3/MIP-1? is a risk factor in the outcome of multiple myeloma (MM), particularly in the development of osteolytic bone disease. This chemokine, highly overexpressed by MM cells, can signal mainly through 2 receptors, CCR1 and CCR5, only 1 of which (CCR1) is responsive to CCL3 in human and mouse osteoclast precursors. CCR1 activation leads to the formation of osteolytic lesions and facilitates tumor growth. Here we show that formation of mature osteoclasts is blocked by the highly potent and selective CCR1 antagonist CCX721, an analog of the clinical compound CCX354. We also show that doses of CCX721 selected to completely inhibit CCR1 produce a profound decrease in tumor burden and osteolytic damage in the murine 5TGM1 model of MM bone disease. Similar effects were observed when the antagonist was used prophylactically or therapeutically, with comparable efficacy to that of zoledronic acid. 5TGM1 cells were shown to express minimal levels of CCR1 while secreting high levels of CCL3, suggesting that the therapeutic effects of CCX721 result from CCR1 inhibition on non-MM cells, most likely osteoclasts and osteoclast precursors. These results provide a strong rationale for further development of CCR1 antagonists for the treatment of MM and associated osteolytic bone disease.

Oyajobi, Babatunde O.; Gupta, Anjana; McCluskey, Brandon; Miao, Shichang; Powers, Jay P.; Seitz, Lisa C.; Wang, Yu; Zeng, Yibin; Zhang, Penglie; Schall, Thomas J.; Jaen, Juan C.

2012-01-01

257

Ovariectomy enhances and estrogen replacement inhibits the activity of bone marrow factors that stimulate prostaglandin production in cultured mouse calvariae.  

PubMed Central

To examine PG production in estrogen deficiency, we studied effects on cultured neonatal mouse calvariae of bone marrow supernatants (MSup) from sham-operated (SHAM), ovariectomized (OVX), or 17 beta-estradiol (OVX+E)-treated mice. MSups were obtained 3 wk after OVX when bone density had decreased significantly. 10-60% MSup increased medium PGE2 and levels of mRNA for inducible and constitutive prostaglandin G/H synthase (PGHS-2 and PGHS-1) and cytosolic phospholipase A2 in calvarial cultures. OVX MSups had twofold greater effects on PGHS-2 and medium PGE2 than other MSups. IL-1 receptor antagonist and anti-IL-1 alpha neutralizing antibody decreased MSup-stimulated PGHS-2 mRNA and PGE2 levels and diminished differences among OVX, sham-operated, and OVX+E groups. In contrast, antibodies to IL-1 beta, IL-6, IL-11, and TNF alpha had little effect. There were no significant differences in IL-1 alpha concentrations or IL-1 alpha mRNA levels in MSups or marrow cells. PGHS-2 mRNA in freshly isolated tibiae from OVX mice was slightly greater than from sham-operated. We conclude that bone marrow factors can increase PG production through stimulation of PGHS-2; that OVX increases and estrogen decreases activity of these factors; and that IL-1 alpha activity, together with additional unknown factors, mediates the differential MSup effects. Images

Kawaguchi, H; Pilbeam, C C; Vargas, S J; Morse, E E; Lorenzo, J A; Raisz, L G

1995-01-01

258

Bone Marrow Stem Cell Origin of Human Breast Cancer Using Transgenic Mouse Models.  

National Technical Information Service (NTIS)

There is emerging evidence that transformed stem cells may be the source of human cancers. We felt that transgenic mouse models were ideally suited to examine this question and proposed to conduct marrow transplant experiments to test whether marrow stem ...

S. H. Barsky

2008-01-01

259

Is bone marrow biopsy always indicated in patients with primary cutaneous marginal zone B-cell lymphoma?  

PubMed

Bone marrow involvement at the time of diagnosis is uncommon in patients with primary cutaneous marginal zone B-cell lymphoma (PCMZL). Moreover, in these patients such involvement is rarely found in isolation on diagnosis. Typically the few patients with PCMZL who have early bone marrow involvement also present secondary nodal or visceral involvement, which is detected by other staging studies (usually computed tomography). In recent years, this has given rise to some debate about whether a bone marrow biopsy should be routinely performed in patients diagnosed with PCMZL in view of the good prognosis and low incidence of bone marrow infiltration and/or extracutaneous involvement in this type of lymphoma. PMID:23954046

Muniesa, C; Hernández-Machín, B

2013-10-01

260

Pharmacologic inhibition of bone resorption prevents cancer-induced osteolysis but enhances soft tissue metastasis in a mouse model of osteolytic breast cancer  

PubMed Central

Osteoprotegerin (OPG) is a secreted member of the TNF receptor superfamily, which binds to the receptor activator of nuclear factor ?B ligand (RANKL) and inhibits osteoclast activity and bone resorption. Systemic administration of recombinant OPG was previously shown to inhibit tumor growth in bone and to prevent cancer-induced osteolysis. In this study, we examined the effect of OPG, when produced locally by breast cancer cells located within bone, using a mouse model of osteolytic breast cancer. MDA-MB-231-TXSA breast cancer cells, tagged with a luciferase reporter gene construct and engineered to overexpress full-length human OPG, were transplanted directly into the tibial marrow cavity of nude mice. Overexpression of OPG by breast cancer cells protected the bone from breast cancer-induced osteolysis and diminished intra-osseous tumor growth but had no effect on extra-skeletal tumor growth. This effect was associated with a significant reduction in the number of osteoclasts that lined the bone surface, resulting in a net increase in bone volume. Despite limiting breast cancer-mediated bone loss, OPG overexpression resulted in a significant increase in the incidence of pulmonary metastasis. Our results demonstrate that inhibition of osteoclastic bone resorption by OPG when secreted locally by tumors in bone may affect the behaviour of cancer cells within the bone microenvironment and their likelihood of spreading and establishing metastasis elsewhere in the body.

ZINONOS, IRENE; LUO, KE-WANG; LABRINIDIS, AGATHA; LIAPIS, VASILIOS; HAY, SHELLEY; PANAGOPOULOS, VASILIOS; DENICHILO, MARK; KO, CHUN-HAY; YUE, GRACE GAR-LEE; LAU, CLARA BIK-SAN; INGMAN, WENDY; PONOMAREV, VLADIMIR; ATKINS, GERALD J.; FINDLAY, DAVID M.; ZANNETTINO, ANDREW C.W.; EVDOKIOU, ANDREAS

2014-01-01

261

Pharmacologic inhibition of bone resorption prevents cancer-induced osteolysis but enhances soft tissue metastasis in a mouse model of osteolytic breast cancer.  

PubMed

Osteoprotegerin (OPG) is a secreted member of the TNF receptor superfamily, which binds to the receptor activator of nuclear factor ?B ligand (RANKL) and inhibits osteoclast activity and bone resorption. Systemic administration of recombinant OPG was previously shown to inhibit tumor growth in bone and to prevent cancer-induced osteolysis. In this study, we examined the effect of OPG, when produced locally by breast cancer cells located within bone, using a mouse model of osteolytic breast cancer. MDA-MB-231-TXSA breast cancer cells, tagged with a luciferase reporter gene construct and engineered to overexpress full-length human OPG, were transplanted directly into the tibial marrow cavity of nude mice. Overexpression of OPG by breast cancer cells protected the bone from breast cancer-induced osteolysis and diminished intra-osseous tumor growth but had no effect on extra-skeletal tumor growth. This effect was associated with a significant reduction in the number of osteoclasts that lined the bone surface, resulting in a net increase in bone volume. Despite limiting breast cancer-mediated bone loss, OPG overexpression resulted in a significant increase in the incidence of pulmonary metastasis. Our results demonstrate that inhibition of osteoclastic bone resorption by OPG when secreted locally by tumors in bone may affect the behaviour of cancer cells within the bone microenvironment and their likelihood of spreading and establishing metastasis elsewhere in the body. PMID:24865346

Zinonos, Irene; Luo, Ke-Wang; Labrinidis, Agatha; Liapis, Vasilios; Hay, Shelley; Panagopoulos, Vasilios; Denichilo, Mark; Ko, Chun-Hay; Yue, Grace Gar-Lee; Lau, Clara Bik-San; Ingman, Wendy; Ponomarev, Vladimir; Atkins, Gerald J; Findlay, David M; Zannettino, Andrew C W; Evdokiou, Andreas

2014-08-01

262

Primary leiomyosarcoma of bone—a clinicopathologic study of 8 uncommon cases with immunohistochemical analysis and clinical outcomes  

Microsoft Academic Search

Primary leiomyosarcoma of bone is a rare and a diagnostically challenging tumor entity. Over a 7-year period, we identified 8 such cases that fulfilled the diagnostic criteria in 6 men and 2 women, with age ranging from 25 to 59 years (mean, 42.7 years). All cases were noted in the lower limbs, including femur and tibia as the commonly involved

Bharat Rekhi; Amrit Kaur; Ajay Puri; Saral Desai; N. A. Jambhekar

2011-01-01

263

Prolonged and ubiquitous inhibition by interferon ? of bone resorption induced by parathyroid hormone-related protein, 1,25-dihydroxyvitamin D 3 , and interleukin 1 in fetal mouse forearm bones  

Microsoft Academic Search

Summary  To investigate the mechanism of the inhibitory effects of interferon-? (IFN-?) on bone resorption, the effects of murine IFN-?\\u000a on45Ca release from prelabeled fetal mouse forearm bones were investigated. Murine IFN-? usually did not affect basal45Ca release but almost completely and equipotently inhibited bone resorption induced by PTH(1-34), PTH-rP(1-34), 1,25(OH)2D3, and interleukin 1 (IL-1). The half-maximal concentration for inhibition of

Yuko Fujii; Kanji Sato; Keizo Kasono; Tomoko Satoh; Toru Fujii; Kazuo Shizume

1990-01-01

264

Host bone marrow-derived IL-12 enhances donor T cell engraftment in a mouse model of bone marrow transplantation  

PubMed Central

Background Donor cell engraftment is critical for the success of allogeneic bone marrow transplants. Graft failure is a result of donor cells either failing to engraft initially or being eliminated at later time points. Donor cell engraftment is facilitated by donor T cells, which eliminate residual host hemato-lymphoid effector cells such as NK cells and T cells. Methods We aimed to explore the role of host hematopoietic cell derived IL-12 on donor cell engraftment in a murine model of BMT. We established radiation chimeras by transplanting C57BL6/J (B6) mice with BM from either congenic B6 mice or IL-12p40 KO mice. These WT ? WT or IL-12 KO ? WT chimeras then underwent a secondary transplant with allogeneic (FVB) BM. Survival, engraftment, donor T cell expansion, cytokine production by donor T cells, as well as expression of stimulatory markers on donor T cells was analyzed. Results Mice whose residual host hematopoietic cells were capable of producing IL-12 had modestly higher survival, higher donor T cell engraftment, and significantly higher donor erythroid engraftment. We have also found that an increased number of donor T cells in IL-12 KO ? WT chimeras have a regulatory phenotype, expressing FoxP3, producing lower levels of TNF-?, higher levels of IL-10, and expressing higher levels of ICOS as well as PD-1 on CD4+ T cells. Conclusions To our knowledge, this is the first report of a beneficial role of IL-12 production by host cells in the context of bone marrow engraftment in a murine model of BMT. These findings support the clinical use of exogenous IL-12 for use in settings where graft failure is of concern.

2014-01-01

265

CXC receptor knockout mice: Characterization of skeletal features and membranous bone healing in the adult mouse  

Microsoft Academic Search

The potential role of CXC chemokines bearing the glu-leu-arg (ELR) motif in bone repair was studied using a cranial defect (CD) model in mice lacking the CXC receptor (mCXCR?\\/? knockout mice), which is homologous to knockout of the human CXC receptor 2 (CXCR2) gene. During the inflammatory stage of bone repair, ELR CXC chemokines are released by inflammatory cells and

David S. Bischoff; Taylor Sakamoto; Kenji Ishida; Nalini S. Makhijani; Helen E. Gruber; Dean T. Yamaguchi

2011-01-01

266

Characterization of a new renal cell carcinoma bone metastasis mouse model  

Microsoft Academic Search

Metastatic bone disease caused by renal cell carcinoma (RCC) occurs frequently and becomes more and more prevalent presumably\\u000a because survival times among patients with disseminated cancers are increasing. Patients with bone metastases from renal cell\\u000a carcinoma suffer from severe pain, nerve compression syndromes and pathologic fractures. Very little is known about the mechanisms\\u000a of skeletal metastases of RCC. Thus, to

Anne Strube; Elizaveta Stepina; Dominik Mumberg; Arne Scholz; Peter Hauff; Sanna-Maria Käkönen

2010-01-01

267

Biocompatibility effects of biologically synthesized graphene in primary mouse embryonic fibroblast cells  

PubMed Central

Due to unique properties and unlimited possible applications, graphene has attracted abundant interest in the areas of nanobiotechnology. Recently, much work has focused on the synthesis and properties of graphene. Here we show that a successful reduction of graphene oxide (GO) using spinach leaf extract (SLE) as a simultaneous reducing and stabilizing agent. The as-prepared SLE-reduced graphene oxide (S-rGO) was characterized by ultraviolet–visible spectroscopy and Fourier transform infrared spectroscopy. Dynamic light scattering technique was used to determine the average size of GO and S-rGO. Scanning electron microscopy and atomic force microscopy images provide clear surface morphological evidence for the formation of graphene. The resulting S-rGO has a mostly single-layer structure, is stable, and has significant water solubility. In addition, the biocompatibility of graphene was investigated using cell viability, leakage of lactate dehydrogenase and alkaline phosphatase activity in primary mouse embryonic fibroblast (PMEFs) cells. The results suggest that the biologically synthesized graphene has significant biocompatibility with PMEF cells, even at a higher concentration of 100 ?g/mL. This method uses a ‘green’, natural reductant and is free of additional stabilizing reagents; therefore, it is an environmentally friendly, simple, and cost-effective method for the fabrication of soluble graphene. This study could open up a promising view for substitution of hydrazine by a safe, biocompatible, and powerful reduction for the efficient deoxygenation of GO, especially in large-scale production and potential biomedical applications.

2013-01-01

268

Alkbh2 protects against lethality and mutation in primary mouse embryonic fibroblasts  

PubMed Central

Alkylating agents modify DNA and RNA forming adducts that disrupt replication and transcription, trigger cell cycle checkpoints and/or initiate apoptosis. If left unrepaired, some of the damage can be cytotoxic and/or mutagenic. In Escherichia coli, the alkylation repair protein B (AlkB) provides one form of resistance to alkylating agents by eliminating mainly 1-methyladenine and 3-methylcytosine, thereby increasing survival and preventing mutation. To examine the biological role of the mammalian AlkB homologs Alkbh2 and Alkbh3, which both have similar enzymatic activities to that of AlkB, we evaluated the survival and mutagenesis of primary Big Blue mouse embryonic fibroblasts (MEFs) that had targeted deletions in the Alkbh2 or Alkbh3 genes. Both Alkbh2- and Alkbh3-deficient MEFs were ~2-fold more sensitive to methyl methanesulfonate (MMS) induced cytotoxicity compared to the wild type control cells. Spontaneous mutant frequencies were similar for the wild type, Alkbh2?/? and Alkbh3?/? MEFs (average-1.3×10?5). However, despite the similar survival of the two mutant MEFs after MMS treatment, only the Alkbh2-deficient MEFs showed a statistically significant increase in mutant frequency compared to wild type MEFs after MMS treatment. Therefore, although both Alkbh2 and Alkbh3 can protect against MMS-induced cell death, only Alkbh2 shows statistically significant protection of MEF DNA against mutations following treatment with this exogenous methylating agent.

Nay, Stephanie L.; Lee, Dong-Hyun; Bates, Steven E.; O'Connor, Timothy R.

2013-01-01

269

Efficacy of phosphatidylinositol-3 kinase inhibitors in a primary mouse model of undifferentiated pleomorphic sarcoma.  

PubMed

Recent advances in sarcoma genomics have identified novel mutations in the PI3K pathway in human sarcomas. Here, we use a mouse model of primary soft-tissue sarcoma for preclinical testing of doxorubicin and inhibitors of the PI3K pathway: BKM120 (PI3K inhibitor) and BEZ235 (a dual PI3K/mTOR inhibitor). Doxorubicin-treated tumors (n = 15) showed a partial response rate of 6.6%, just as the majority of human sarcomas do not respond to doxorubicin. Treatment with BKM120 elicited a partial response in 50% of tumors (n = 10), which was also seen in combination with doxorubicin (n = 10). Additionally, BKM120 treatment produced a robust delay in tumor growth kinetics. BEZ235-treated tumors (n = 9) showed a complete response rate of 11.1%. Combining BEZ235 with doxorubicin (n = 10) increased the complete response rate to 50% (P = 0.035). These studies demonstrate that PI3K pathway inhibition is a viable and attractive target for soft-tissue sarcomas. PMID:22619567

Kim, Suzy; Dodd, Rebecca D; Mito, Jeffrey K; Ma, Yan; Kim, Yongbaek; Riedel, Richard F; Kirsch, David G

2012-01-01

270

Transcriptomics analysis of primary mouse thymocytes exposed to bis(tri-n-butyltin)dioxide (TBTO).  

PubMed

The biocide bis(tri-n-butyltin)oxide (TBTO) causes thymus atrophy in rodents and is toxic to many cell types of which thymocytes are the most sensitive. To obtain insight in the mechanisms of action of TBTO, we exposed primary mouse thymocytes in vitro for 3, 6 and 11 h to 0.1, 0.5, 1 and 2 ?M TBTO. Subsequently, the cells were subjected to whole-genome gene expression profiling. Biological interpretation of the gene expression data revealed that TBTO affects a wide range of processes. Cell proliferation related genes were downregulated by all treatments except for 3 and 6 h 0.5 ?M TBTO which upregulated these genes. Treatment with TBTO resulted in upregulation of genes involved in endoplasmatic reticulum (ER) stress, NFkB and TNF? pathways, and genes involved in DNA damage, p53 signaling and apoptosis. Remarkably, TBTO also increased the expression of genes that are known to be upregulated during T cell activation or during negative selection of thymocytes. The effect of TBTO on expression of genes involved in ER stress and apoptosis was confirmed by qPCR. Induction of the T cell activation response was corroborated by demonstrating that TBTO exposure resulted in translocation of NFAT to the nucleus, which is an essential event for T cell activation. PMID:22434021

van Kol, Sandra W M; Hendriksen, Peter J M; van Loveren, Henk; Peijnenburg, Ad

2012-06-14

271

Ovarian abnormalities in a mouse model of fragile X primary ovarian insufficiency.  

PubMed

FMR1 premutation (PM) alleles have 55-200 CGG·CCG-repeats in their 5' UTR. PM carriers are at risk of fragile X-associated tremor and ataxia syndrome (FXTAS). Females are also at risk for FX primary ovarian insufficiency (FXPOI). PM pathology is generally attributed to deleterious properties of transcripts with long CGG-tracts. For FXPOI, hormone changes suggest a reduced residual follicle pool. Whether this is due to a smaller than normal original follicle pool or an increased rate of follicle depletion is unclear. A FX-PM mouse the authors generated with 130 CGG·CCG-repeats in the endogenous Fmr1 gene recapitulates features of FXTAS. Here the authors demonstrate that the gross development of the ovary and the establishment of the primordial follicle pool is normal in these mice. However, these animals show a faster loss of follicles of all follicle classes, suggesting that the problem is intrinsic to the ovary. In addition, many oocytes show aberrant nuclear accumulation of FMRP and elevated levels of ubiquitination. Furthermore, PM follicles are smaller and have fewer granulosa cells (GCs) than normal. Thus, these animals have ovarian abnormalities involving both the oocytes and GCs that may shed light on the molecular basis of FXPOI in humans. PMID:22470123

Hoffman, Gloria E; Le, Wei Wei; Entezam, Ali; Otsuka, Noriyuki; Tong, Zhi-Bin; Nelson, Lawrence; Flaws, Jodi A; McDonald, John H; Jafar, Sanjeeda; Usdin, Karen

2012-06-01

272

Ovarian Abnormalities in a Mouse Model of Fragile X Primary Ovarian Insufficiency  

PubMed Central

FMR1 premutation (PM) alleles have 55–200 CGG·CCG-repeats in their 5? UTR. PM carriers are at risk of fragile X–associated tremor and ataxia syndrome (FXTAS). Females are also at risk for FX primary ovarian insufficiency (FXPOI). PM pathology is generally attributed to deleterious properties of transcripts with long CGG-tracts. For FXPOI, hormone changes suggest a reduced residual follicle pool. Whether this is due to a smaller than normal original follicle pool or an increased rate of follicle depletion is unclear. A FX-PM mouse the authors generated with 130 CGG·CCG-repeats in the endogenous Fmr1 gene recapitulates features of FXTAS. Here the authors demonstrate that the gross development of the ovary and the establishment of the primordial follicle pool is normal in these mice. However, these animals show a faster loss of follicles of all follicle classes, suggesting that the problem is intrinsic to the ovary. In addition, many oocytes show aberrant nuclear accumulation of FMRP and elevated levels of ubiquitination. Furthermore, PM follicles are smaller and have fewer granulosa cells (GCs) than normal. Thus, these animals have ovarian abnormalities involving both the oocytes and GCs that may shed light on the molecular basis of FXPOI in humans.

Le, Wei Wei; Entezam, Ali; Otsuka, Noriyuki; Tong, Zhi-Bin; Nelson, Lawrence; Flaws, Jodi A.; McDonald, John H.; Jafar, Sanjeeda; Usdin, Karen

2012-01-01

273

Secondary Aneurysmal Bone Cystic Change of the Chondroblastoma, Mistaken for a Primary Aneurysmal Bone Cyst in the Patella  

PubMed Central

A 29-year-old woman complained of a 3-month history of left knee pain without trauma history. X-ray showed a well-defined osteolytic lesion with a sclerotic margin in the patella and magnetic resonance imaging showed T1-low and T2-high signal intensity with different fluid level. Our impression was an aneurysmal bone cyst. At surgery, the lesion was a blood-filled cystic cavity, surrounded by a gray or brownish tissue. Hemorrhagic soft tissues with recognizable bone fragments were observed. Curettage and autogenous bone graft was done. Microscopically, sheets of tumor cells were intermingled with some areas of eosinophilic chondroid matrix. The tumor cells showed oval-shaped nuclei with moderate eosinophilic cytoplasm. Several multinucleated giant cells and blood filled cystic cavities were observed. The final diagnosis was a chondroblastoma with a secondary aneurysmal bone cyst. At the post-operative 1.5-year follow-up, grafted bones were well incorporated radiographically and there were no recurrent evidence or any other abnormal symptoms.

Chung, Jin Wha

2014-01-01

274

Secondary aneurysmal bone cystic change of the chondroblastoma, mistaken for a primary aneurysmal bone cyst in the patella.  

PubMed

A 29-year-old woman complained of a 3-month history of left knee pain without trauma history. X-ray showed a well-defined osteolytic lesion with a sclerotic margin in the patella and magnetic resonance imaging showed T1-low and T2-high signal intensity with different fluid level. Our impression was an aneurysmal bone cyst. At surgery, the lesion was a blood-filled cystic cavity, surrounded by a gray or brownish tissue. Hemorrhagic soft tissues with recognizable bone fragments were observed. Curettage and autogenous bone graft was done. Microscopically, sheets of tumor cells were intermingled with some areas of eosinophilic chondroid matrix. The tumor cells showed oval-shaped nuclei with moderate eosinophilic cytoplasm. Several multinucleated giant cells and blood filled cystic cavities were observed. The final diagnosis was a chondroblastoma with a secondary aneurysmal bone cyst. At the post-operative 1.5-year follow-up, grafted bones were well incorporated radiographically and there were no recurrent evidence or any other abnormal symptoms. PMID:24639947

Chung, Jin Wha; Lee, Hwa Sung

2014-03-01

275

Primary cilium-dependent mechanosensing is mediated by adenylyl cyclase 6 and cyclic AMP in bone cells  

PubMed Central

Primary cilia are chemosensing and mechanosensing organelles that regulate remarkably diverse processes in a variety of cells. We previously showed that primary cilia play a role in mediating mechanosensing in bone cells through an unknown mechanism that does not involve extracellular Ca2+-dependent intracellular Ca2+ release, which has been implicated in all other cells that transduce mechanical signals via the cilium. Here, we identify a molecular mechanism linking primary cilia and bone cell mechanotransduction that involves adenylyl cyclase 6 (AC6) and cAMP. Intracellular cAMP was quantified in MLO-Y4 cells exposed to dynamic flow, and AC6 and primary cilia were inhibited using RNA interference. When exposed to flow, cells rapidly (<2 min) and transiently decreased cAMP production in a primary cilium-dependent manner. RT-PCR revealed differential expression of the membrane-bound isoforms of adenylyl cyclase, while immunostaining revealed one, AC6, preferentially localized to the cilium. Further studies showed that decreases in cAMP in response to flow were dependent on AC6 and Gd3+-sensitive channels but not intracellular Ca2+ release and that this response mediated flow-induced COX-2 gene expression. The signaling events identified provide important details of a novel early mechanosensing mechanism in bone and advances our understanding of how signal transduction occurs at the primary cilium.—Kwon, R. Y., Temiyasathit, S., Tummala, P., Quah, C. C., Jacobs, C. R. Primary cilium-dependent mechanosensing is mediated by adenylyl cyclase 6 and cyclic AMP in bone cells.

Kwon, Ronald Y.; Temiyasathit, Sara; Tummala, Padmaja; Quah, Clarence C.; Jacobs, Christopher R.

2010-01-01

276

Systemic Delivery of Oncolytic Adenoviruses Targeting Transforming Growth Factor-? Inhibits Established Bone Metastasis in a Prostate Cancer Mouse Model  

PubMed Central

Abstract We have examined whether Ad.sT?RFc and TAd.sT?RFc, two oncolytic viruses expressing soluble transforming growth factor-? receptor II fused with human Fc (sTGF?RIIFc), can be developed to treat bone metastasis of prostate cancer. Incubation of PC-3 and DU-145 prostate tumor cells with Ad.sT?RFc and TAd.sT?RFc produced sTGF?RIIFc and viral replication; sTGF?RIIFc caused inhibition of TGF-?-mediated SMAD2 and SMAD3 phosphorylation. Ad(E1-).sT?RFc, an E1– adenovirus, produced sTGF?RIIFc but failed to replicate in tumor cells. To examine the antitumor response of adenoviral vectors, PC-3-luc cells were injected into the left heart ventricle of nude mice. On day 9, mice were subjected to whole-body bioluminescence imaging (BLI). Mice bearing hind-limb tumors were administered viral vectors via the tail vein on days 10, 13, and 17 (2.5×1010 viral particles per injection per mouse, each injection in a 0.1-ml volume), and subjected to BLI and X-ray radiography weekly until day 53. Ad.sT?RFc, TAd.sT?RFc, and Ad(E1-).sT?RFc caused significant inhibition of tumor growth; however, Ad.sT?RFc was the most effective among all the vectors. Only Ad.sT?RFc and TAd.sT?RFc inhibited tumor-induced hypercalcemia. Histomorphometric and synchrotron micro-computed tomographic analysis of isolated bones indicated that Ad.sT?RFc induced significant reduction in tumor burden, osteoclast number, and trabecular and cortical bone destruction. These studies suggest that Ad.sT?RFc and TAd.sT?RFc can be developed as potential new therapies for prostate cancer bone metastasis.

Hu, Zebin; Gupta, Janhavi; Zhang, Zhenwei; Gerseny, Helen; Berg, Arthur; Chen, Yun Ju; Zhang, Zhiling; Du, Hongyan; Brendler, Charles B.; Xiao, Xianghui; Pienta, Kenneth J.; Guise, Theresa; Lee, Chung; Stern, Paula H.; Stock, Stuart

2012-01-01

277

Effect of peripherally administered leptin antagonist on whole body metabolism and bone microarchitecture and biomechanical properties in the mouse.  

PubMed

Leptin's in vivo effect on the rodent skeleton depends on the model used and the mode of administration. Superactive mouse leptin antagonist (SMLA) was produced and then pegylated (PEG) to prolong and enhance its in vivo activity. We blocked leptin signaling by injecting this antagonist peripherally into normal mice at various time points and studied their metabolic and skeletal phenotypes. Subcutaneous PEG-SMLA injections into 4-wk-old female C57BL/6J mice increased weight gain and food consumption significantly after only 1 mo, and the effect lasted for the 3 mo of the experiment, proving its central inhibiting activity. Mice showed a significant increase in serum glucose, cholesterol, triglycerides, insulin, and HOMA-IR throughout the experiment. Quantification of gene expression in "metabolic" tissues also indicated the development of insulin resistance. Bone analyses revealed a significant increase in trabecular and cortical parameters measured in both the lumbar vertebrae and tibiae in PEG-SMLA-treated mice in the 1st and 3rd months as well as a significant increase in tibia biomechanical parameters. Interestingly, 30 days of treatment with the antagonist in older mice (aged 3 and 6 mo) affected body weight and eating behavior, just as they had in the 1-mo-old mice, but had no effect on bone parameters, suggesting that leptin's effect on bones, either directly or through its obesogenic effect, is dependent upon stage of skeletal development. This potent and reversible antagonist enabled us to study leptin's in vivo role in whole body and bone metabolism and holds potential for future therapeutic use in diseases involving leptin signaling. PMID:24169045

Solomon, Gili; Atkins, Ayelet; Shahar, Ron; Gertler, Arieh; Monsonego-Ornan, Efrat

2014-01-01

278

Targeting A-type K(+) channels in primary sensory neurons for bone cancer pain in a rat model.  

PubMed

Cancer pain is one of the most severe types of chronic pain, and the most common cancer pain is bone cancer pain. The treatment of bone cancer pain remains a clinical challenge. Here, we report firstly that A-type K(+) channels in dorsal root ganglion (DRG) are involved in the neuropathy of rat bone cancer pain and are a new target for diclofenac, a nonsteroidal anti-inflammatory drug that can be used for therapy for this distinct pain. There are dynamically functional changes of the A-type K(+) channels in DRG neurons during bone cancer pain. The A-type K(+) currents that mainly express in isolectin B4-positive small DRG neurons are increased on post-tumor day 14 (PTD 14), then faded but still remained at a higher level on PTD 21. Correspondingly, the expression levels of A-type K(+) channel Kv1.4, Kv3.4, and Kv4.3 showed time-dependent changes during bone cancer pain. Diclofenac enhances A-type K(+) currents in the DRG neurons and attenuates bone cancer pain in a dose-dependent manner. The analgesic effect of diclofenac can be reversed or prevented by A-type K(+) channel blocker 4-AP or pandinotoxin-K?, also by siRNA targeted against rat Kv1.4 or Kv4.3. Repeated diclofenac administration decreased soft tissue swelling adjacent to the tumor and attenuated bone destruction. These results indicate that peripheral A-type K(+) channels were involved in the neuropathy of rat bone cancer pain. Targeting A-type K(+) channels in primary sensory neurons may provide a novel mechanism-based therapeutic strategy for bone cancer pain. PMID:22188869

Duan, Kai-Zheng; Xu, Qian; Zhang, Xiao-Meng; Zhao, Zhi-Qi; Mei, Yan-Ai; Zhang, Yu-Qiu

2012-03-01

279

In vivo cyclic compression causes cartilage degeneration and subchondral bone changes in mouse tibiae  

PubMed Central

Objectives Alterations in the mechanical loading environment in joints may have both beneficial and detrimental effects on articular cartilage and subchondral bone and subsequently influence the development of osteoarthritis (OA). We used an in vivo tibial loading model to investigate the adaptive responses of cartilage and bone to mechanical loading and to assess the influence of load level and duration. Methods We applied cyclic compression of 4.5 and 9.0N peak loads to the left tibia via the knee joint of adult (26-week-old) C57Bl/6 male mice for 1, 2, and 6 weeks. Only 9.0N loading was utilized in young (10-week-old) mice. The changes in articular cartilage and subchondral bone were analyzed by histology and microcomputed tomography. Results Loading promoted cartilage damage in both age groups, with increased damage severity dependent upon the duration of loading. Metaphyseal bone mass increased in the young mice, but not in the adult mice, whereas epiphyseal cancellous bone mass decreased with loading in both young and adult mice. Articular cartilage thickness decreased, and subchondral cortical bone thickness increased in the posterior tibial plateau in both age groups. Both age groups developed periarticular osteophytes at the tibial plateau in response to the 9.0N load, but no osteophyte formation occurred in adult mice subjected to 4.5N peak loading. Conclusion This non-invasive loading model permits dissection of temporal and topographical changes in cartilage and bone and will enable investigation of the efficacy of treatment interventions targeting joint biomechanics or biological events that promote OA onset and progression.

Ko, Frank C.; Dragomir, Cecilia; Plumb, Darren A.; Goldring, Steven R.; Wright, Timothy M.; Goldring, Mary B.; van der Meulen, Marjolein C.H.

2013-01-01

280

Genetic modification of mouse bone marrow by lentiviral vector-mediated delivery of hypoxanthine-Guanine phosphoribosyltransferase short hairpin RNA confers chemoprotection against 6-thioguanine cytotoxicity.  

PubMed

We have recently developed a novel and highly efficient strategy that exclusively uses the purine analog 6-thioguanine (6TG) for both pretransplantation conditioning and post-transplantation chemoselection of hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient bone marrow (BM). In a mouse BM transplantation model, combined 6TG preconditioning and in vivo chemoselection consistently achieved >95% engraftment of HPRT-deficient donor BM and long-term reconstitution of histologically and immunophenotypically normal hematopoiesis in both primary and secondary recipients, without significant toxicity and in the absence of any other cytotoxic conditioning regimen. To translate this strategy for combined 6TG conditioning and chemoselection into a clinically feasible approach, it is necessary to develop methods for genetic modification of normal hematopoietic stem cells (HSC) to render them HPRT-deficient and thus 6TG-resistant. Here we investigated a strategy to reduce HPRT expression and thereby confer protection against 6TG myelotoxicity to primary murine BM cells by RNA interference (RNAi). Accordingly, we constructed and validated a lentiviral gene transfer vector expressing short-hairpin RNA (shRNA) that targets the murine HPRT gene. Our results showed that lentiviral vector-mediated delivery of HPRT-targeted shRNA could achieve effective and long-term reduction of HPRT expression. Furthermore, in both an established murine cell line as well as in primary murine BM cells, lentiviral transduction with HPRT-targeted shRNA was associated with enhanced resistance to 6TG cytotoxicity in vitro. Hence this represents a translationally feasible method to genetically engineer HSC for implementation of 6TG-mediated preconditioning and in vivo chemoselection. PMID:23769104

Hacke, K; Treger, J A; Bogan, B T; Schiestl, R H; Kasahara, N

2013-06-01

281

VCP Associated Inclusion Body Myopathy and Paget Disease of Bone Knock-In Mouse Model Exhibits Tissue Pathology Typical of Human Disease  

Microsoft Academic Search

Dominant mutations in the valosin containing protein (VCP) gene cause inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD). We have generated a knock-in mouse model with the common R155H mutation. Mice demonstrate progressive muscle weakness starting approximately at the age of 6 months. Histology of mutant muscle showed progressive vacuolization of myofibrils and centrally located

Mallikarjun Badadani; Angèle Nalbandian; Giles D. Watts; Jouni Vesa; Masashi Kitazawa; Hailing Su; Jasmin Tanaja; Eric Dec; Douglas C. Wallace; Jogeshwar Mukherjee; Vincent Caiozzo; Matthew Warman; Virginia E. Kimonis

2010-01-01

282

Compound heterozygous loss of Ext1 and Ext2 is sufficient for formation of multiple exostoses in mouse ribs and long bones  

Microsoft Academic Search

Multiple Hereditary Exostoses (MHE) syndrome is caused by haploinsufficiency in Golgi-associated heparan sulfate polymerases EXT1 or EXT2 and is characterized by formation of exostoses next to growing long bones and other skeletal elements. Recent mouse studies have indicated that formation of stereotypic exostoses requires a complete loss of Ext expression, suggesting that a similar local loss of EXT function may

Beverly M. Zak; Manuela Schuksz; Eiki Koyama; Christina Mundy; Dan E. Wells; Yu Yamaguchi; Maurizio Pacifici; Jeffrey D. Esko

2011-01-01

283

Cross-species Bone Marrow Transplantation: Evidence for Tolerance Induction, Stem Cell Engraftment, and Maturation ofT Lymphocytes in a Xenogeneic Stromal Environment (Rat --> Mouse)  

Microsoft Academic Search

Summary Transplantation of untreated F344 rat bone marrow into irradiated B10 mouse recipients (non- TCD F344 -> B10) to produce fully xenogeneic chimeras resulted in stable xenogeneic lymphoid chimerism, ranging from 82% to 97% rat . Survival of animals was excellent, without evidence for GVH disease. The specificity of tolerance which resulted was highly donor-specific ; MHC disparate third partymouse

Suzanne T. Ildstad; Sherry M. Wren; Sallie S. Boggs; Mary L. Hronakes; Fausto Vecchini; M. Van den Brink

284

Pathological Fractures in Primary Non-Hodgkin's Lymphoma of the Bone: A Case Series with Review of the Literature  

PubMed Central

Primary non-Hodgkin’s lymphoma of bone (PLB) is a rare entity. Patients generally present with localized bone pain and, less frequently, soft-tissue swelling or a palpable mass. Pathological fracture of the proximal femur and proximal humerus secondary to soft-tissue tumours is well documented in the literature; however, lymphomas presenting primarily at these sites with pathological fracture is unusual. A review of the world literature shows that the incidence of skeletal manifestation from NHL is less than 5%, and in all these cases, bony involvement was reported many years after presentation of the primary cancer. Histopathologically, PLB usually represents diffuse large B-cell lymphoma. We report our experience with two cases of Primary non-Hodgkin’s lymphoma of proximal femur and proximal humerus with pathological fracture and their management.

Siddiqui, Yasir Salam; Khan, Abdul Qayyum; Sherwani, MKA

2013-01-01

285

Skp2 promotes adipocyte differentiation via a p27 Kip1-independent mechanism in primary mouse embryonic fibroblasts  

Microsoft Academic Search

Skp2, the substrate-binding subunit of an SCF ubiquitin ligase complex, is a key regulator of cell cycle progression that targets substrates for degradation by the 26S proteasome. We have now shown that ablation of Skp2 in primary mouse embryonic fibroblasts (MEFs) results both in impairment of adipocyte differentiation and in the accumulation of the cyclin-dependent kinase inhibitor p27Kip1, a principal

Mitsuru Okada; Tamon Sakai; Takehiro Nakamura; Mimi Tamamori-Adachi; Shigetaka Kitajima; Yasushi Matsuki; Eijiro Watanabe; Ryuji Hiramatsu; Hiroshi Sakaue; Masato Kasuga

2009-01-01

286

Skp2 promotes adipocyte differentiation via a p27{sup Kip1}-independent mechanism in primary mouse embryonic fibroblasts  

Microsoft Academic Search

Skp2, the substrate-binding subunit of an SCF ubiquitin ligase complex, is a key regulator of cell cycle progression that targets substrates for degradation by the 26S proteasome. We have now shown that ablation of Skp2 in primary mouse embryonic fibroblasts (MEFs) results both in impairment of adipocyte differentiation and in the accumulation of the cyclin-dependent kinase inhibitor p27{sup Kip1}, a

Mitsuru Okada; Tamon Sakai; Takehiro Nakamura; Mimi Tamamori-Adachi; Shigetaka Kitajima; Yasushi Matsuki; Eijiro Watanabe; Ryuji Hiramatsu; Hiroshi Sakaue; Masato Kasuga

2009-01-01

287

Improvement of Cardiac Function in Mouse Myocardial Infarction after Transplantation of Epigenetically-Modified Bone Marrow Progenitor Cells  

PubMed Central

Objective To study usefulness of bone marrow progenitor cells (BPCs) epigenetically altered by chromatin modifying agents in mediating heart repair after myocardial infarction in mice. Methods and Results We tested the therapeutic efficacy of bone marrow progenitor cells treated with the clinically-used chromatin modifying agents Trichostatin A (TSA, histone deacetylase inhibitor) and 5Aza-2-deoxycytidine (Aza, DNA methylation inhibitor) in a mouse model of acute myocardial infarction (AMI). Treatment of BPCs with Aza and TSA induced expression of pluripotent genes Oct4, Nanog, Sox2, and thereafter culturing these cells in defined cardiac myocyte-conditioned medium resulted in their differentiation into cardiomyocyte progenitors and subsequently into cardiac myocytes. Their transition was deduced by expression of repertoire of markers: Nkx2.5, GATA4, cardiotroponin T, cardiotroponin I, ?-sarcomeric actinin, Mef2c and MHC-?. We observed that the modified BPCs had greater AceH3K9 expression and reduced histone deacetylase1 (HDAC1) and lysine-specific demethylase1 (LSD1) expression compared to untreated BPCs, characteristic of epigenetic changes. Intra-myocardial injection of modified BPCs after AMI in mice significantly improved left ventricular function. These changes were ascribed to differentiation of the injected cells into cardiomyocytes and endothelial cells. Conclusion Treatment of BPCs with Aza and TSA converts BPCs into multipotent cells, which can then be differentiated into myocyte progenitors. Transplantation of these modified progenitor cells into infarcted mouse hearts improved left ventricular function secondary to differentiation of cells in the niche into myocytes and endothelial cells.

Rajasingh, Johnson; Thangavel, Jayakumar; Siddiqui, Mohammad R.; Gomes, Ignatius; Gao, Xiao-pei; Kishore, Raj; Malik, Asrar B.

2011-01-01

288

Bone Marrow Stromal Cells Modulate Mouse ENT1 Activity and Protect Leukemia Cells from Cytarabine Induced Apoptosis  

PubMed Central

Background Despite a high response rate to chemotherapy, the majority of patients with acute myeloid leukemia (AML) are destined to relapse due to residual disease in the bone marrow (BM). The tumor microenvironment is increasingly being recognized as a critical factor in mediating cancer cell survival and drug resistance. In this study, we propose to identify mechanisms involved in the chemoprotection conferred by the BM stroma to leukemia cells. Methods Using a leukemia mouse model and a human leukemia cell line, we studied the interaction of leukemia cells with the BM microenvironment. We evaluated in vivo and in vitro leukemia cell chemoprotection to different cytotoxic agents mediated by the BM stroma. Leukemia cell apoptosis was assessed by flow cytometry and western blotting. The activity of the equilibrative nucleoside transporter 1 (ENT1), responsible for cytarabine cell incorporation, was investigated by measuring transport and intracellular accumulation of 3H-adenosine. Results Leukemia cell mobilization from the bone marrow into peripheral blood in vivo using a CXCR4 inhibitor induced chemo-sensitization of leukemia cells to cytarabine, which translated into a prolonged survival advantage in our mouse leukemia model. In vitro, the BM stromal cells secreted a soluble factor that mediated significant chemoprotection to leukemia cells from cytarabine induced apoptosis. Furthermore, the BM stromal cell supernatant induced a 50% reduction of the ENT1 activity in leukemia cells, reducing the incorporation of cytarabine. No protection was observed when radiation or other cytotoxic agents such as etoposide, cisplatin and 5-fluorouracil were used. Conclusion The BM stroma secretes a soluble factor that significantly protects leukemia cells from cytarabine-induced apoptosis and blocks ENT1 activity. Strategies that modify the chemo-protective effects mediated by the BM microenvironment may enhance the benefit of conventional chemotherapy for patients with AML.

Macanas-Pirard, Patricia; Leisewitz, Andrea; Broekhuizen, Richard; Cautivo, Kelly; Barriga, Francisco M.; Leisewitz, Francisco; Gidi, Victoria; Riquelme, Erick; Montecinos, Viviana P.; Swett, Pilar; Besa, Pelayo; Ramirez, Pablo; Ocqueteau, Mauricio; Kalergis, Alexis M.; Holt, Matthew; Rettig, Michael; DiPersio, John F.; Nervi, Bruno

2012-01-01

289

Primary non-Hodgkin's lymphoma of the bladder with bone marrow involvement.  

PubMed

Involvement of the lower urinary tract by advanced non-Hodgkin's lymphoma (NHL) has been reported in up to 13% of cases, but primary NHL of the urinary bladder is very rare. A 35-year-old man was admitted to our hospital with a chief complaint of gross hematuria with left flank pain on April 12, 2001. Cystoscopy revealed an edematous broad-based mass on the left lateral wall of the bladder, and transurethral biopsy showed NHL, diffuse large B-cell type. Abdomino-pelvic CT scan demonstrated left-side hydronephrosis and hydroureter with left proximal ureter infiltration and thickening of the left lateral wall of the bladder with perivesical fat infiltration without lymph node enlargement. Full-scale staging work-up revealed the bone marrow as the solely involved site. The lesions of the bladder and left urinary tract were nearly completely regressed after two cycles of systemic cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy with simultaneous restoration of urinary function. PMID:12760267

Oh, Kil Chan; Zang, Dae Young

2003-03-01

290

Binocular input coincidence mediates critical period plasticity in the mouse primary visual cortex.  

PubMed

Classical studies on the development of ocular dominance (OD) organization in primary visual cortex (V1) have revealed a postnatal critical period (CP), during which visual inputs between the two eyes are most effective in shaping cortical circuits through synaptic competition. A brief closure of one eye during CP caused a pronounced shift of response preference of V1 neurons toward the open eye, a form of CP plasticity in the developing V1. However, it remains unclear what particular property of binocular inputs during CP is responsible for mediating this experience-dependent OD plasticity. Using whole-cell recording in mouse V1, we found that visually driven synaptic inputs from the two eyes to binocular cells in layers 2/3 and 4 became highly coincident during CP. Enhancing cortical GABAergic transmission activity by brain infusion with diazepam not only caused a precocious onset of the high coincidence of binocular inputs and OD plasticity in pre-CP mice, but rescued both of them in dark-reared mice, suggesting a tight link between coincident binocular inputs and CP plasticity. In Thy1-ChR2 mice, chronic disruption of this binocular input coincidence during CP by asynchronous optogenetic activation of retinal ganglion cells abolished the OD plasticity. Computational simulation using a feed-forward network model further suggests that the coincident inputs could mediate this CP plasticity through a homeostatic synaptic learning mechanism with synaptic competition. These results suggest that the high-level correlation of binocular inputs is a hallmark of the CP of developing V1 and serves as neural substrate for the induction of OD plasticity. PMID:24553935

Chen, Xiao-Jing; Rasch, Malte J; Chen, Guang; Ye, Chang-Quan; Wu, Si; Zhang, Xiao-Hui

2014-02-19

291

Neurogenic and Neurotrophic Effects of BDNF Peptides in Mouse Hippocampal Primary Neuronal Cell Cultures  

PubMed Central

The level of brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, is down regulated in Alzheimer’s disease (AD), Parkinson’s disease (PD), depression, stress, and anxiety; conversely the level of this neurotrophin is increased in autism spectrum disorders. Thus, modulating the level of BDNF can be a potential therapeutic approach for nervous system pathologies. In the present study, we designed five different tetra peptides (peptides B-1 to B-5) corresponding to different active regions of BDNF. These tetra peptides were found to be non-toxic, and they induced the expression of neuronal markers in mouse embryonic day 18 (E18) primary hippocampal neuronal cultures. Additionally, peptide B-5 induced the expression of BDNF and its receptor, TrkB, suggesting a positive feedback mechanism. The BDNF peptides induced only a moderate activation (phosphorylation at Tyr 706) of the TrkB receptor, which could be blocked by the Trk’s inhibitor, K252a. Peptide B-3, when combined with BDNF, potentiated the survival effect of this neurotrophin on H2O2-treated E18 hippocampal cells. Peptides B-3 and B-5 were found to work as partial agonists and as partial antagonists competing with BDNF to activate the TrkB receptor in a dose-dependent manner. Taken together, these results suggest that the described BDNF tetra peptides are neurotrophic, can modulate BDNF signaling in a partial agonist/antagonist way, and offer a novel therapeutic approach to neural pathologies where BDNF levels are dysregulated.

Cardenas-Aguayo, Maria del Carmen; Kazim, Syed Faraz; Grundke-Iqbal, Inge; Iqbal, Khalid

2013-01-01

292

Protection of primary neurons and mouse brain from Alzheimer's pathology by molecular tweezers  

PubMed Central

Alzheimer’s disease is a devastating cureless neurodegenerative disorder affecting >35 million people worldwide. The disease is caused by toxic oligomers and aggregates of amyloid ? protein and the microtubule-associated protein tau. Recently, the Lys-specific molecular tweezer CLR01 has been shown to inhibit aggregation and toxicity of multiple amyloidogenic proteins, including amyloid ? protein and tau, by disrupting key interactions involved in the assembly process. Following up on these encouraging findings, here, we asked whether CLR01 could protect primary neurons from Alzheimer’s disease-associated synaptotoxicity and reduce Alzheimer’s disease–like pathology in vivo. Using cell culture and brain slices, we found that CLR01 effectively inhibited synaptotoxicity induced by the 42-residue isoform of amyloid ? protein, including ?80% inhibition of changes in dendritic spines density and long-term potentiation and complete inhibition of changes in basal synaptic activity. Using a radiolabelled version of the compound, we found that CLR01 crossed the mouse blood–brain barrier at ?2% of blood levels. Treatment of 15-month-old triple-transgenic mice for 1 month with CLR01 resulted in a decrease in brain amyloid ? protein aggregates, hyperphosphorylated tau and microglia load as observed by immunohistochemistry. Importantly, no signs of toxicity were observed in the treated mice, and CLR01 treatment did not affect the amyloidogenic processing of amyloid ? protein precursor. Examining induction or inhibition of the cytochrome P450 metabolism system by CLR01 revealed minimal interaction. Together, these data suggest that CLR01 is safe for use at concentrations well above those showing efficacy in mice. The efficacy and toxicity results support a process-specific mechanism of action of molecular tweezers and suggest that these are promising compounds for developing disease-modifying therapy for Alzheimer’s disease and related disorders.

Attar, Aida; Ripoli, Cristian; Riccardi, Elisa; Maiti, Panchanan; Li Puma, Domenica D.; Liu, Tingyu; Hayes, Jane; Jones, Mychica R.; Lichti-Kaiser, Kristin; Yang, Fusheng; Gale, Greg D.; Tseng, Chi-hong; Tan, Miao; Xie, Cui-Wei; Straudinger, Jeffrey L.; Klarner, Frank-Gerrit; Schrader, Thomas; Frautschy, Sally A.; Grassi, Claudio

2012-01-01

293

The Mouse Primary Visual Cortex Is a Site of Production and Sensitivity to Estrogens  

PubMed Central

The classic female estrogen, 17?-estradiol (E2), has been repeatedly shown to affect the perceptual processing of visual cues. Although gonadal E2 has often been thought to influence these processes, the possibility that central visual processing may be modulated by brain-generated hormone has not been explored. Here we show that estrogen-associated circuits are highly prevalent in the mouse primary visual cortex (V1). Specifically, we cloned aromatase, a marker for estrogen-producing neurons, and the classic estrogen receptors (ERs) ER? and ER?, as markers for estrogen-responsive neurons, and conducted a detailed expression analysis via in-situ hybridization. We found that both monocular and binocular V1 are highly enriched in aromatase- and ER-positive neurons, indicating that V1 is a site of production and sensitivity to estrogens. Using double-fluorescence in-situ hybridization, we reveal the neurochemical identity of estrogen-producing and -sensitive cells in V1, and demonstrate that they constitute a heterogeneous neuronal population. We further show that visual experience engages a large population of aromatase-positive neurons and, to a lesser extent, ER-expressing neurons, suggesting that E2 levels may be locally regulated by visual input in V1. Interestingly, acute episodes of visual experience do not affect the density or distribution of estrogen-associated circuits. Finally, we show that adult mice dark-reared from birth also exhibit normal distribution of aromatase and ERs throughout V1, suggesting that the implementation and maintenance of estrogen-associated circuits is independent of visual experience. Our findings demonstrate that the adult V1 is a site of production and sensitivity to estrogens, and suggest that locally-produced E2 may shape visual cortical processing.

Majewska, Ania K.; Pinaud, Raphael

2011-01-01

294

The mouse primary visual cortex is a site of production and sensitivity to estrogens.  

PubMed

The classic female estrogen, 17?-estradiol (E2), has been repeatedly shown to affect the perceptual processing of visual cues. Although gonadal E2 has often been thought to influence these processes, the possibility that central visual processing may be modulated by brain-generated hormone has not been explored. Here we show that estrogen-associated circuits are highly prevalent in the mouse primary visual cortex (V1). Specifically, we cloned aromatase, a marker for estrogen-producing neurons, and the classic estrogen receptors (ERs) ER? and ER?, as markers for estrogen-responsive neurons, and conducted a detailed expression analysis via in-situ hybridization. We found that both monocular and binocular V1 are highly enriched in aromatase- and ER-positive neurons, indicating that V1 is a site of production and sensitivity to estrogens. Using double-fluorescence in-situ hybridization, we reveal the neurochemical identity of estrogen-producing and -sensitive cells in V1, and demonstrate that they constitute a heterogeneous neuronal population. We further show that visual experience engages a large population of aromatase-positive neurons and, to a lesser extent, ER-expressing neurons, suggesting that E2 levels may be locally regulated by visual input in V1. Interestingly, acute episodes of visual experience do not affect the density or distribution of estrogen-associated circuits. Finally, we show that adult mice dark-reared from birth also exhibit normal distribution of aromatase and ERs throughout V1, suggesting that the implementation and maintenance of estrogen-associated circuits is independent of visual experience. Our findings demonstrate that the adult V1 is a site of production and sensitivity to estrogens, and suggest that locally-produced E2 may shape visual cortical processing. PMID:21647225

Jeong, Jin-Kwon; Tremere, Liisa A; Burrows, Kaiping; Majewska, Ania K; Pinaud, Raphael

2011-01-01

295

Intracellular cholesterol-binding proteins enhance HDL-mediated cholesterol uptake in cultured primary mouse hepatocytes  

PubMed Central

A major gap in our knowledge of rapid hepatic HDL cholesterol clearance is the role of key intracellular factors that influence this process. Although the reverse cholesterol transport pathway targets HDL to the liver for net elimination of free cholesterol from the body, molecular details governing cholesterol uptake into hepatocytes are not completely understood. Therefore, the effects of sterol carrier protein (SCP)-2 and liver fatty acid-binding protein (L-FABP), high-affinity cholesterol-binding proteins present in hepatocyte cytosol, on HDL-mediated free cholesterol uptake were examined using gene-targeted mouse models, cultured primary hepatocytes, and 22-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-amino]-23,24-bisnor-5-cholen-3?-ol (NBD-cholesterol). While SCP-2 overexpression enhanced NBD-cholesterol uptake, counterintuitively, SCP-2/SCP-x gene ablation also 1) enhanced the rapid molecular phase of free sterol uptake detectable in <1 min and initial rate and maximal uptake of HDL free cholesterol and 2) differentially enhanced free cholesterol uptake mediated by the HDL3, rather than the HDL2, subfraction. The increased HDL free cholesterol uptake was not due to increased expression or distribution of the HDL receptor [scavenger receptor B1 (SRB1)], proteins regulating SRB1 [postsynaptic density protein (PSD-95)/Drosophila disk large tumor suppressor (dlg)/tight junction protein (ZO1) and 17-kDa membrane-associated protein], or other intracellular cholesterol trafficking proteins (steroidogenic acute response protein D, Niemann Pick C, and oxysterol-binding protein-related proteins). However, expression of L-FABP, the single most prevalent hepatic cytosolic protein that binds cholesterol, was upregulated twofold in SCP-2/SCP-x null hepatocytes. Double-immunogold electron microscopy detected L-FABP sufficiently close to SRB1 for direct interaction, similar to SCP-2. These data suggest a role for L-FABP in HDL cholesterol uptake, a finding confirmed with SCP-2/SCP-x/L-FABP null mice and hepatocytes. Taken together, these results suggest that L-FABP, particularly in the absence of SCP-2, plays a significant role in HDL-mediated cholesterol uptake in cultured primary hepatocytes.

Storey, Stephen M.; McIntosh, Avery L.; Huang, Huan; Landrock, Kerstin K.; Martin, Gregory G.; Landrock, Danilo; Payne, H. Ross; Atshaves, Barbara P.; Kier, Ann B.

2012-01-01

296

Chewing ameliorates chronic mild stress-induced bone loss in senescence-accelerated mouse (SAMP8), a murine model of senile osteoporosis.  

PubMed

Chronic mild stress is a risk factor for osteoporosis and chewing inhibits the stress response. We examined the effect of chewing on chronic stress-induced bone loss and bone microstructural deterioration in mice. The senescence-accelerated mouse strain P8 (SAMP8) was randomly divided into control, stress, and stress with chewing groups of fifteen animals each. Mice in the stress and stress with chewing groups were placed in a ventilated restraint tube for 60minutes, twice a day for 4weeks. The restrained mice were simultaneously subjected daily to one of the following stressors: water immersion, physical shaking and flashing lights. Mice in the stress with chewing group were allowed to chew a wooden stick during the experimental period. After the experiment, the bone response was evaluated using quantitative micro computed tomography, bone histomorphometry, and biochemical markers. Exposure of SAMP8 mice to chronic stress resulted in significant increase of the blood corticosterone and noradrenaline levels, and adrenal weight. The bone resorption was activated and the bone formation was suppressed. Trabecular bone volume and trabecular number were decreased in both the vertebra and distal femur of the stress group. Chewing under chronic stress prevented the increase in the blood corticosterone and noradrenaline levels, attenuated the reduced bone formation and increased bone resorption, improved the trabecular bone loss and bone microstructural deterioration induced by chronic mild stress. These findings indicate that chewing can ameliorate chronic stress-induced bone loss in SAMP8 mice. Thus, chewing may represent a useful method preventing and/or treating chronic stress-related osteoporosis. PMID:24607548

Furuzawa, Manabu; Chen, Huayue; Fujiwara, Shu; Yamada, Kumiko; Kubo, Kin-Ya

2014-07-01

297

INFLUENCE OF LEPTIN ON THE EARLY PHASES OF BONE FORMATION DURING MOUSE SKELETON ORGANOGENESIS  

Microsoft Academic Search

Leptin is a 16-kDa hormone, primarily secreted by adipose tissue, which controls body weight through its effects on food intake and energy expenditure by negative feedback at the hypothalamic nuclei. Leptin is now known to have actions in the immune system, reproduction, development, haemopoiesis, angiogenesis and, most recently, in bone metabolism (1). Concerning the skeleton, it seems to be involved

Rivasi Marianna; Benelli Augusta; Ferretti Marzia; Benincasa Marta; Bertoni Laura; Cavani Francesco; Palumbo Carla

2007-01-01

298

Differential Expression of Surface Markers in Mouse Bone Marrow Mesenchymal Stromal Cell Subpopulations with Distinct Lineage Commitment  

PubMed Central

Bone marrow mesenchymal stromal cells (BM MSCs) represent a heterogeneous population of progenitors with potential for generation of skeletal tissues. However the identity of BM MSC subpopulations is poorly defined mainly due to the absence of specific markers allowing in situ localization of those cells and isolation of pure cell types. Here, we aimed at characterization of surface markers in mouse BM MSCs and in their subsets with distinct differentiation potential. Using conditionally immortalized BM MSCs we performed a screening with 176 antibodies and high-throughput flow cytometry, and found 33 markers expressed in MSCs, and among them 3 were novel for MSCs and 13 have not been reported for MSCs from mice. Furthermore, we obtained clonally derived MSC subpopulations and identified bipotential progenitors capable for osteo- and adipogenic differentiation, as well as monopotential osteogenic and adipogenic clones, and thus confirmed heterogeneity of MSCs. We found that expression of CD200 was characteristic for the clones with osteogenic potential, whereas SSEA4 marked adipogenic progenitors lacking osteogenic capacity, and CD140a was expressed in adipogenic cells independently of their efficiency for osteogenesis. We confirmed our observations in cell sorting experiments and further investigated the expression of those markers during the course of differentiation. Thus, our findings provide to our knowledge the most comprehensive characterization of surface antigens expression in mouse BM MSCs to date, and suggest CD200, SSEA4 and CD140a as markers differentially expressed in distinct types of MSC progenitors.

Rostovskaya, Maria; Anastassiadis, Konstantinos

2012-01-01

299

Activation of the A(3) adenosine receptor suppresses superoxide production and chemotaxis of mouse bone marrow neutrophils.  

PubMed

Adenosine is formed in injured/ischemic tissues, where it suppresses the actions of essentially all cells of the immune system. Most of the anti-inflammatory actions of adenosine have been attributed to signaling through the G(s) protein-coupled A(2A) adenosine receptor (AR). Here, we report that the A(3)AR is highly expressed in murine neutrophils isolated from bone marrow. Selective activation of the A(3)AR with (2S,3S,4R,5R)-3-amino-5-[6-(2,5-dichlorobenzylamino)purin-9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide (CP-532,903) potently inhibited mouse bone marrow neutrophil superoxide generation and chemotaxis induced by various activating agents. The selectivity of CP-532,903 was confirmed in assays using neutrophils obtained from A(2A)AR and A(3)AR gene "knockout" mice. In a model of thioglycollate-induced inflammation, treating mice with CP-532,903 inhibited recruitment of leukocytes into the peritoneum by specifically activating the A(3)AR. Collectively, our findings support the theory that the A(3)AR contributes to the anti-inflammatory actions of adenosine on neutrophils and provide a potential mechanistic explanation for the efficacy of A(3)AR agonists in animal models of inflammation (i.e., inhibition of neutrophil-mediated tissue injury). PMID:18583455

van der Hoeven, Dharini; Wan, Tina C; Auchampach, John A

2008-09-01

300

Effect of L-carnitine on erythroid colony formation in mouse bone marrow cells  

Microsoft Academic Search

Background. L-Carnitine can alleviate uraemic anaemia in haemodialysis patients by improving erythrocyte membrane functions or erythropoiesis, which are depressed under uraemic conditions. L-Carnitine and palmitoyl-L-carnitine were reported to increase the formation of colony-forming unit-erythroid (CFU-E) colonies in cultures of fetal mouse liver cells, an effect that depended on the concentration of palmitoyl-L- carnitine but not of L-carnitine. In this study,

Yukika Kitamura; Kazunori Satoh; Takashi Satoh; Manabu Takita; Akihiro Matsuura

301

Comparative Analysis of Temporal and Dose-Dependent TCDD-Elicited Gene Expression in Human, Mouse, and Rat Primary Hepatocytes  

PubMed Central

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)–elicited time- and dose-dependent differential gene expression was compared in human, mouse, and rat primary hepatocytes. Comprehensive time course (10 nM TCDD or dimethyl sulfoxide vehicle control for 1, 2, 4, 8, 12, 24, and 48h) studies identified 495, 2305, and 711 differentially expressed orthologous genes in human, mouse, and rat hepatocytes, respectively. However, only 16 orthologs were differentially expressed across all three species, with the majority of orthologs exhibiting species-specific expression (399 human, 2097 mouse, and 533 rat), consistent with species-specific expression reported in other in vitro and in vivo comparative studies. TCDD also elicited the dose-dependent induction of 397 human, 100 mouse, and 443 rat genes at 12h and 615 human, 426 mouse, and 314 rat genes at 24h. Comparable EC50 values were obtained for AhR battery genes including Cyp1a1 (0.1 nM human, 0.05 nM mouse, 0.08 nM rat at 24h) and Tiparp (0.97 nM human, 0.63 nM mouse, 0.14 nM rat at 12h). Overrepresented functions and pathways included amino acid metabolism in humans, immune response in mice, and energy homeostasis in rats. Differentially expressed genes functionally associated with lipid transport, processing, and metabolism were overrepresented in all three species but exhibited species-specific expression consistent with the induction of hepatic steatosis in mice but not in rats following a single oral gavage of TCDD. Furthermore, human primary hepatocytes showed lipid accumulation following 48h of treatment with TCDD, suggesting that AhR-mediated steatosis in mice more closely resembles human hepatic fat accumulation compared with that in rats. Collectively, these results suggest that species-specific gene expression profiles mediate the species-specific effects of TCDD despite the conservation of the AhR and its signaling mechanism.

Zacharewski, Timothy R.

2013-01-01

302

Fetal and postnatal mouse bone tissue contains more calcium than is present in hydroxyapatite  

Microsoft Academic Search

It has been shown for developing enamel and zebrafish fin that hydroxyapatite (HA) is preceded by an amorphous precursor, motivating us to examine the mineral development in mammalian bone, particularly femur and tibia of fetal and young mice. Mineral particle thickness and arrangement were characterized by (synchrotron) small-angle X-ray scattering (SAXS) combined with wide-angle X-ray diffraction (WAXD) and X-ray fluorescence

C. Lange; C. Li; I. Manjubala; W. Wagermaier; J. Kühnisch; M. Kolanczyk; S. Mundlos; P. Knaus; P. Fratzl

2011-01-01

303

The applicability of the Greulich & Pyle Atlas for bone age assessment in primary school-going children of Karachi, Pakistan  

PubMed Central

Objective: To assess the degree of applicability of bone age calculated by Greulich & Pyle Atlas in estimation of chronological age for therapeutic and medico legal purposes. Methods: Two Hundred and Twenty children (139 males, 81 females) between ages of 56 and 113 months (4.5 to 9.5 years) were randomly selected from 4 primary schools of Shireen Jinnah & Clifton, Karachi. Digital images of hand and wrist radiographs were obtained by a computed radiography at Ziauddin Hospital Clifton. Bone ages were computed using Greulich & Pyle Atlas by radiologists at Ziauddin Hospital, North Nazimabad, Karachi. Results: On average, the Greulich & Pyle Atlas underestimates chronological age by 6.65 ± 13.47 months in females and 15.78 ± 12.83 months in males (p-values < 0.001). High correlation was found between chronological age and bone age in both genders (Females r=0.778; p-value< 0.001, Males r=0.816; p-value < 0.001). Conclusion: Bone age calculated by Greulich & Pyle Atlas should not be used for estimating chronological age in children of ages 56-113 months in situations where high accuracy is required (e.g. medicolegal cases). However, serial measurements of bone age by this atlas can be used in management of growth related endocrine disorders in these children.

Manzoor Mughal, Arsalan; Hassan, Nuzhat; Ahmed, Anwar

2014-01-01

304

Culture conditions allow selection of different mesenchymal progenitors from adult mouse bone marrow.  

PubMed

The use of adult stem cells in tissue engineering approaches will benefit from the establishment of culture conditions that allow the expansion and maintenance of cells with stem cell-like activity and high differentiation potential. In the field of adult stem cells, bone marrow stromal cells (BMSCs) are promising candidates. In the present study, we define, for the first time, conditions for optimizing the yields of cultures enriched for specific progenitors of bone marrow. Using four distinct culture conditions, supernatants from culture of bone fragments, marrow stroma cell line MS-5, embryonic fibroblast cell line NIH3T3, and a cocktail of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF), we isolated four different sub-populations of murine BMSCs (mBMSCs). These cells express a well-known marker of undifferentiated embryonic stem cells (Nanog) and show interesting features in immunophenotype, self-renewal ability, and differentiation potency. In particular, using NIH3T3 conditioned medium, we obtained cells that showed impairment in osteogenic and chondrogenic differentiation while retaining high adipogenic potential during passages. Our results indicate that the choice of the medium used for isolation and expansion of mBMSCs is important for enriching the culture of desired specific progenitors. PMID:19298168

Esposito, Maria Teresa; Di Noto, Rosa; Mirabelli, Peppino; Gorrese, Marisa; Parisi, Silvia; Montanaro, Donatella; Del Vecchio, Luigi; Pastore, Lucio

2009-09-01

305

Talbot-defocus multiscan tomography using the synchrotron X-ray microscope to study the lacuno-canalicular network in mouse bone.  

PubMed

The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes. We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network. The resultant pair of tomograms was precisely aligned with each other, allowing evaluation of mineral density in the vicinity of each osteocyte lacuna and canaliculus over the entire thickness of the cortical bone. Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level. PMID:23761853

Nango, Nobuhito; Kubota, Shogo; Takeuchi, Akihisa; Suzuki, Yoshio; Yashiro, Wataru; Momose, Atsushi; Matsuo, Koichi

2013-06-01

306

Talbot-defocus multiscan tomography using the synchrotron X-ray microscope to study the lacuno-canalicular network in mouse bone  

PubMed Central

The three-dimensional network of lacunae and canaliculi that regulates metabolism in bone contains osteocytes and their dendritic processes. We constructed a synchrotron radiation X-ray microscope for sequential tomography of mouse tibia first by using a Talbot interferometer to detect the degree of bone mineralization and then by using absorption contrast under a slightly defocused setting to enhance outline contrast thereby visualizing structures of the osteocyte lacuno-canalicular network. The resultant pair of tomograms was precisely aligned with each other, allowing evaluation of mineral density in the vicinity of each osteocyte lacuna and canaliculus over the entire thickness of the cortical bone. Thus, multiscan microscopic X-ray tomography is a powerful tool for analyzing bone mineralization in relation to the lacuno-canalicular network at the submicron resolution level.

Nango, Nobuhito; Kubota, Shogo; Takeuchi, Akihisa; Suzuki, Yoshio; Yashiro, Wataru; Momose, Atsushi; Matsuo, Koichi

2013-01-01

307

Primary hyperparathyroidism with severe bone disease: osteitis fibrosa cystica vs. fibrous dysplasia. Case report and review of the literature  

Microsoft Academic Search

Primary hyperparathyroidism (HPT) is associated with generalized skeletal changes, its full-blown osseous manifestations known as osteitis fibrosa cystica. Fibrous dysplasia (FD), a benign bone disorder, is differentiated from generalized fibrocystic disease caused by hyperparathyroidism. The classic triad of McCune–Albright syndrome includes polyostotic FD, patchy skin pigmentation, and sexual precocity. Other associated endocrinopathies are hyperthyroidism, Cushing's syndrome, acromegaly, and HPT. We

Maria João Leitão; Lu??s Cuña; Nuno Pinheiro; V??tor Coelho; Mário Oliveira; João Mascarenhas Araújo

1999-01-01

308

The Use of Structural Allograft in Primary and Revision Knee Arthroplasty with Bone Loss  

PubMed Central

Bone loss around the knee in the setting of total knee arthroplasty remains a difficult and challenging problem for orthopaedic surgeons. There are a number of options for dealing with smaller and contained bone loss; however, massive segmental bone loss has fewer options. Small, contained defects can be treated with cement, morselized autograft/allograft or metal augments. Segmental bone loss cannot be dealt with through simple addition of cement, morselized autograft/allograft, or metal augments. For younger or higher demand patients, the use of allograft is a good option as it provides a durable construct with high rates of union while restoring bone stock for future revisions. Older patients, or those who are low demand, may be better candidates for a tumour prosthesis, which provides immediate ability to weight bear and mobilize.

Kuchinad, Raul A.; Garbedian, Shawn; Rogers, Benedict A.; Backstein, David; Safir, Oleg; Gross, Allan E.

2011-01-01

309

Effect of bone morphogenetic protein-4 (BMP-4) on cardiomyocyte differentiation from mouse embryonic stem cell.  

PubMed

The present study was designed to evaluate the effect of BMP-4 on mouse embryonic stem cells (ESCs)-derived cardiomyocyte. Cardiac differentiation of the mouse ESCs was initiated by embryoid bodies (EBs) formation in hanging drops, transfer of EBs to the suspension culture and then plating onto gelatin-coated tissue culture plates. BMP-4 was added to culture medium throughout the suspension period. Cultures were observed daily with an inverted microscope for the appearance of contracting clusters. At the early, intermediate and terminal stages of differentiation, the choronotropic responses of cardiomyocytes to cardioactive drugs were assessed, and the cardiomyocytes immunostained for cardiac troponin I, desmin, alpha-actinin and nebulin. The contracting clusters were isolated for ultrastructural evaluation, at day 14 after plating. Moreover, total RNA extracted from contracting EBs of early and terminal stages of differentiation were examined for oct-4, alpha- and beta-myosin heavy chain, myosin light chain-2V and atrial natriuretic factor expression. The BMP-4 treatment resulted in a decrease in the percent of beating EBs and the percent of developing cardiomyocytes per EBs. As a whole, the chronotropic responses of beating cardiac clusters to cardioactive drugs in control group were better than BMP-4 treated group. The cardiomyocytes of both groups were positive immunostained for applied antibodies except for nebulin. Moreover, in the BMP-4 treated group, the ultrastructural characteristics and cardiac-specific genes expression were all retarded in the terminal stage of cardiomyocytes development. In conclusion, BMP-4 had an inhibitory effect on cardiomyocyte differentiation from the mouse ESCs in terms of ultrastructural characteristics, genes expression and functional properties. PMID:17156864

Taha, Masoumeh Fakhr; Valojerdi, Mojtaba Rezazadeh; Mowla, Seyed Javad

2007-08-01

310

Columnar metaplasia in a surgical mouse model of gastro-esophageal reflux disease is not derived from bone marrow-derived cell.  

PubMed

The incidence of esophageal adenocarcinoma has increased in the last 25 years. Columnar metaplasia in Barrett's mucosa is assumed to be a precancerous lesion for esophageal adenocarcinoma. However, the induction process of Barrett's mucosa is still unknown. To analyze the induction of esophageal columnar metaplasia, we established a mouse gastro-esophageal reflux disease (GERD) model with associated development of columnar metaplasia in the esophagus. C57BL/6 mice received side-to-side anastomosis of the esophagogastric junction with the jejunum, and mice were killed 10, 20, and 40 weeks after operation. To analyze the contribution of bone marrow-derived cells to columnar metaplasia in this surgical GERD model, some mice were transplanted with GFP-marked bone marrow after the operation. Seventy-three percent of the mice (16/22) showed thickened mucosa in esophagus and 41% of mice (9/22) developed columnar metaplasia 40 weeks after the operation with a mortality rate of 4%. Bone marrow-derived cells were not detected in columnar metaplastic epithelia. However, scattered epithelial cells in the thickened squamous epithelia in regions of esophagitis did show bone marrow derivation. The results demonstrate that reflux induced by esophago-jejunostomy in mice leads to the development of columnar metaplasia in the esophagus. However, bone marrow-derived cells do not contribute directly to columnar metaplasia in this mouse model. PMID:23734763

Aikou, Susumu; Aida, Junko; Takubo, Kaiyo; Yamagata, Yukinori; Seto, Yasuyuki; Kaminishi, Michio; Nomura, Sachiyo

2013-09-01

311

The accuracy of computer-assisted primary mandibular reconstruction with vascularized bone flaps: iliac crest bone flap versus osteomyocutaneous fibula flap  

PubMed Central

Background The intention of mandibular reconstruction is to restore the complex anatomy with maximum possible functionality and high accuracy. The aim of this study was to evaluate the accuracy of computer-assisted surgery in primary mandibular reconstruction with an iliac crest bone flap compared with an osteomyocutaneous fibula flap. Materials and methods Preoperative computed tomography data of the mandible and the iliac crest or fibula donor site were imported into a specific surgical planning software program. Surgical guides were manufactured using a rapid prototyping technique for translating the virtual plan, including information on the transplant dimensions and shape, into real-time surgery. Using postoperative computed tomography scans and an automatic surface-comparison algorithm, the actual postoperative situation was compared with the preoperative virtual simulation. Results The actual flap position showed a mean difference from the virtual plan of 2.43 mm (standard deviation [SD] ±1.26) and a surface deviation of 39% <2 mm and 15% <1 mm for the iliac crest bone flap, and a mean difference of 2.18 mm (SD ±1.93) and a surface deviation of 60% <2 mm and 37% <1 mm for the osteomyocutaneous fibula flap. The position of the neomandible reconstructed with an osteomyocutaneous fibula flap indicated a mean difference from the virtual plan of 1.25 mm (SD ±1.31) and a surface deviation of 82% <2 mm and 57% <1 mm, in contrast to a mean difference of 1.68 mm (SD ±1.25) and a surface deviation of 63% <2 mm and 38% <1 mm for the neomandible after reconstruction with an iliac crest bone flap. For shape analysis, a similarly high accuracy could be calculated for both flaps. Conclusion Virtual surgical planning is an effective method for mandibular reconstruction with vascularized bone flaps, and can help to restore the anatomy of the mandible with high accuracy in position and shape. It seems that primary mandibular reconstruction with the osteomyocutaneous fibula flap is more accurate compared with the vascularized iliac crest bone flap.

Modabber, Ali; Ayoub, Nassim; Mohlhenrich, Stephan Christian; Goloborodko, Evgeny; Sonmez, Tolga Taha; Ghassemi, Mehrangiz; Loberg, Christina; Lethaus, Bernd; Ghassemi, Alireza; Holzle, Frank

2014-01-01

312

SCE frequencies induced by ethanol, tequila and brandy in mouse bone marrow cells in vivo.  

PubMed

The genotoxicity of ethanol, tequila and brandy was evaluated by scoring the frequency of sister chromatid exchanges (SCE) and determining the values of the average generation time (AGT). We studied four dosages of each substance i.p. inoculated into mice. The cytogenetic analysis was performed in bone marrow cells. The results showed that all three substances were weak genotoxicants. Tequila showed the strongest response followed by brandy and ethanol. None of them modified the cell proliferation kinetics as demonstrated by the AGT results. PMID:8427015

Piña Calva, A; Madrigal-Bujaidar, E

1993-01-01

313

Caveolin-1 and migration of bone-marrow derived cells in the mouse eye.  

PubMed

Bone marrow derived cells (BMDCs) can be found in almost every tissue showing a distinct turnover and density. Since caveolin-1 regulates junction-associated proteins in endothelial and epithelial cells, its role for BMDC was investigated in the eyes of caveolin-1 knock-out mice transplanted with GFP-marked BMDC. Distribution and turnover of BMDC in connective tissues (cornea, iris, ciliary body and choroid) was not altered. The absence of caveolin-1, however, caused a significant decrease of BMDC turnover in cornea epithelium, ciliary epithelium, and in the retina. This finding emphasizes an important, hitherto unknown role of caveolin-1 in neuronal and epithelial tissues. PMID:23896585

May, Christian-Albrecht

2013-12-01

314

Effects of prostaglandin F2 alpha on bone formation and resorption in cultured neonatal mouse calvariae: Role of prostaglandin E2 production  

SciTech Connect

Although most studies show that prostaglandin E2 (PGE2) is the most potent and effective of the prostanoids in bone, recent data in cell culture suggest that PGF2 alpha may have unique effects, particularly on cell replication. The present study was undertaken to compare the effects of PGF2 alpha and PGE2 in cultured neonatal mouse parietal bones by simultaneous measurement of bone resorption as release of previously incorporated 45Ca, bone formation as incorporation of (3H)proline into collagenase-digestible (CDP) and noncollagen protein, and DNA synthesis as incorporation of (3H)thymidine. PGF2 alpha was less effective than PGE2 as a stimulator of bone resorption, and its effects were partially inhibited by indomethacin and markedly inhibited by glucocorticoids. In contrast, the resorptive response to PGE2 was unaffected by indomethacin and only partially inhibited by cortisol. PGF2 alpha had little effect on bone formation, in contrast to the biphasic effect of PGE2, which inhibited labeling of CDP in the absence of cortisol and stimulated CDP labeling in the presence of cortisol. PGF2 alpha increased thymidine incorporation into DNA, but the effect was smaller than that of PGE2 and was inhibited by indomethacin. These observations suggested that PGF2 alpha might act in part by stimulating PGE2 production. By RIA, PGE2 concentrations were increased in the medium of bones treated with PGF2 alpha, and this increase was blocked by indomethacin. By HPLC, bones prelabeled with (3H)arachidonic acid showed an increase in labeled PGE2 release, and RIA showed an increase in PGE2 after PGF2 alpha treatment. These results indicate that PGF2 alpha is a relatively weak agonist in bone compared to PGE2 and that some of the effects of PGF2 alpha on bone resorption, formation, and cell replication may be mediated by an increase in endogenous PGE2 production.

Raisz, L.G.; Alander, C.B.; Fall, P.M.; Simmons, H.A. (Univ. of Connecticut Health Center, Farmington (USA))

1990-02-01

315

Efficient expansion of mesenchymal stem cells from mouse bone marrow under hypoxic conditions.  

PubMed

To realize the therapeutic potential of mesenchymal stem cells (MSCs), a large number of high-quality MSCs isolated from different species, such as mouse, were acquired for preclinical animal studies. Surprisingly, isolation and purification of mouse MSCs (mMSCs) is arduous because of the low frequency of MSCs and contamination of haematopoietic cells in culture. We have developed a method based on low density and hypoxic culture to isolate and expand mMSCs from different strains, including BALB/c, C57BL/6J, FVB/N and DBA/2. The cells from all of the strains expanded more rapidly when plated at low density in hypoxic culture compared with normoxic culture. These cells expressed CD44, CD105, CD29 and Sca-1 markers but not CD11b, CD34, CD45 and CD31 markers. Moreover, they were able to differentiate along osteoblastic, adipocytic and chondrocytic lineages. In conclusion, we have developed a robust method for isolation and expansion of mMSCs by combining low-density culture with hypoxic culture. PMID:22623422

Yew, Tu-Lai; Chang, Ming-Chau; Hsu, Yuan-Tong; He, Fan-Yu; Weng, Wen-Hui; Tsai, Chih-Chien; Chiu, Fang-Yao; Hung, Shih-Chieh

2013-12-01

316

Primary bone carcinosarcoma of the fibula with chondrosarcoma and squamous cell carcinoma components  

PubMed Central

Carcinosarcoma is defined as a malignant neoplasm that is composed of both carcinomatous and sarcomatous components. The occurrence of carcinosarcoma in the bone is extremely rare. In this report, we describe the third documented de novo case of carcinosarcoma of the bone. A 59-year-old Japanese female presented with a painful tumor in her right lower leg. Plane radiography revealed an osteolytic destructive lesion with periosteal reaction and mineralization in the right fibula. Resection of the fibula tumor was performed under a clinical diagnosis of chondrosarcoma. Histopathological study revealed that the tumor was comprised of three components. The main component was proliferation of small round to short spindle cells (approximately 50%), and the remaining components were chondrosarcoma (30%) and squamous cell carcinoma (20%). Immunohistochemically, SOX9 was expressed in the small round to spindle cells and chondrosarcoma component, and p63 and p40 were expressed in all three components. Accordingly, an ultimate diagnosis of carcinosarcoma of the bone was made. The clinicopathological analysis of carcinosarcoma of the bone revealed that this type of tumor affects the middle-aged to elderly persons and occurs in the long bone. All three de novo cases had chondrosarcoma and squamous cell carcinoma components. One of the 3 patients died of the disease. The histogenesis of carcinosarcoma of the bone remains a matter of controversy, although a multpotential stem cell theory has been proposed. Additional studies are required to clarify the clinical behavior and histogenesis of carcinosarcoma of the bone.

Ishida, Mitsuaki; Kodama, Narihito; Takemura, Yoshinori; Iwai, Muneo; Yoshida, Keiko; Kagotani, Akiko; Matsusue, Yoshitaka; Okabe, Hidetoshi

2013-01-01

317

Functional and Transcriptomic Recovery of Infarcted Mouse Myocardium Treated with Bone Marrow Mononuclear Cells  

PubMed Central

Although bone marrow-derived mononuclear cells (BMNC) have been extensively used in cell therapy for cardiac diseases, little mechanistic information is available to support reports of their efficacy. To address this shortcoming, we compared structural and functional recovery and associated global gene expression profiles in post-ischaemic myocardium treated with BMNC transplantation. BMNC suspensions were injected into cardiac scar tissue 10 days after experimental myocardial infarction. Six weeks later, mice undergoing BMNC therapy were found to have normalized antibody repertoire and improved cardiac performance measured by ECG, treadmill exercise time and echocardiography. After functional testing, gene expression profiles in cardiac tissue were evaluated using high-density oligonucleotide arrays. Expression of more than 18% of the 11981 quantified unigenes was significantly altered in the infarcted hearts. BMNC therapy restored expression of 2099 (96.2%) of the genes that were altered by infarction but led to altered expression of 286 other genes, considered to be a side effect of the treatment. Transcriptional therapeutic efficacy, a metric calculated using a formula that incorporates both recovery and side effect of treatment, was 73%. In conclusion, our results confirm a beneficial role for bone marrow-derived cell therapy and provide new information on molecular mechanisms operating after BMNC transplantation on post ischemic heart failure in mice.

Lachtermacher, Stephan; Esporcatte, Bruno L. B.; da Silva de Azevedo Fortes, Fabio; Rocha, Nazareth Novaes; Montalvao, Fabricio; Costa, Patricia C.; Belem, Luciano; Rabischoffisky, Arnaldo; Neto, Hugo C. C. Faria; Vasconcellos, Rita; Iacobas, Dumitru A.; Iacobas, Sanda; Spray, David C.; Thomas, Neil M.; Goldenberg, Regina C. S.; de Carvalho, Antonio C. Campos

2011-01-01

318

IL-17 is involved in bone resorption in mouse periapical lesions.  

PubMed

Periapical lesions are induced by bacterial infection of the dental pulp and result in destruction of the surrounding alveolar bone. Although various immunological studies concerning periapical bone resorption have been reported, the role of cytokines in the formation of periapical lesions remains unclear. In this study, the role of IL-17A in periapical lesions in mice was investigated. Normal C57BL/6, IFN-gamma(-/-), TNF-alpha(-/-), and IL-17A(-/-) mice were subjected to pulp exposure and infected with Prevotella intermedia (ATCC25611) and Porphyromonas gingivalis (ATCC33277) in the mandibular first molar. Periapical lesions were determined by muCT on day 21 after infection, and 3D visual construction was performed using 3D picture quantification software. The expression of IL-17A mRNA in periapical lesions was determined by the RT-PCR and real-time RT-PCR method. Periapical lesions developed in wild-type, IFN-gamma(-/-), and TNF-alpha(-/-) mice after infection with P. intermedia and P. gingivalis. However, periapical lesions were not observed in IL-17A(-/-) mice. The expression of IL-17A mRNA was significantly induced in periapical lesions of wild-type mice after infection. These results suggest that IL-17A, but not IFN-gamma or TNF-alpha, plays an important role in the formation of periapical lesions. PMID:19457170

Oseko, Fumishige; Yamamoto, Toshiro; Akamatsu, Yuki; Kanamura, Narisato; Iwakura, Yoichiro; Imanishi, Jiro; Kita, Masakazu

2009-05-01

319

?-Adrenergic Agonist and Antagonist Regulation of Autophagy in HepG2 Cells, Primary Mouse Hepatocytes, and Mouse Liver  

PubMed Central

Autophagy recently has been shown to be involved in normal hepatic function and in pathological conditions such as non-alcoholic fatty liver disease. Adrenergic signalling also is an important regulator of hepatic metabolism and function. However, currently little is known about the potential role of adrenergic signaling on hepatic autophagy, and whether the ?-adrenergic receptor itself may be a key regulator of autophagy. To address these issues, we investigated the actions of the ?2-adrenergic receptor agonist, clenbuterol on hepatic autophagy. Surprisingly, we found that clenbuterol stimulated autophagy and autophagic flux in hepatoma cells, primary hepatocytes and in vivo. Similar effects also were observed with epinephrine treatment. Interestingly, propranolol caused a late block in autophagy in the absence and presence of clenbuterol, both in cell culture and in vivo. Thus, our results demonstrate that the ?2- adrenergic receptor is a key regulator of hepatic autophagy, and that the ?-blocker propranolol can independently induce a late block in autophagy.

Farah, Benjamin L.; Sinha, Rohit A.; Wu, Yajun; Singh, Brijesh K.; Zhou, Jin; Bay, Boon-Huat; Yen, Paul M.

2014-01-01

320

Mycobacterium avium Infection Induces H-Ferritin Expression in Mouse Primary Macrophages by Activating Toll-Like Receptor 2  

PubMed Central

Important for both host and pathogen survivals, iron is a key factor in determining the outcome of an infectious process. Iron with-holding, including sequestration inside tissue macrophages, is considered an important strategy to fight infection. However, for intra-macrophagic pathogens, such as Mycobacterium avium, host defence may depend on intracellular iron sequestration mechanisms. Ferritin, the major intracellular iron storage protein, plays a critical role in this process. In the current study, we studied ferritin expression in mouse bone marrow-derived macrophages upon infection with M. avium. We found that H-ferritin is selectively increased in infected macrophages, through an up-regulation of gene transcription. This increase was mediated by the engagement of Toll like receptor-2, and was independent of TNF-alpha or nitric oxide production. The formation of H-rich ferritin proteins and the consequent iron sequestration may be an important part of the panoply of antimicrobial mechanisms of macrophages.

Silva-Gomes, Sandro; Bouton, Cecile; Silva, Tania; Santambrogio, Paolo; Rodrigues, Pedro; Appelberg, Rui; Gomes, Maria Salome

2013-01-01

321

The production of nitric oxide and prostaglandin E(2) by primary bone cells is shear stress dependent.  

PubMed

Loading-induced flow of interstitial fluid through the lacuno-canalicular network is a likely signal for bone cell adaptive responses. However, the nature of the stimulus that activates the cell is debated. Candidate stimuli include wall shear stress, streaming potentials, and chemotransport. We have addressed the nature of the flow-derived cell stimulus by comparing variations in fluid transport with variations in wall shear stress, using nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production as a parameter of bone cell activation. Adult mouse long bone cell cultures were treated for 15min with or without pulsating fluid flow using the following regimes: Low PFF, mean flow rate 0.20 cm(3)/s, 3 Hz, shear stress 0.4+/-0.12 Pa; Medium PFF, 0.33 cm(3)/s, 5 Hz, 0.6+/-0.27 Pa; and High PFF, 0.63 cm(3)/s, 9Hz, 1.2+/-0.37 Pa. In some Low PFF experiments, 2.8% neutral dextran (mol. wt. 4.98x10(4)) was added to the flow medium to increase the viscosity, thereby increasing the wall shear stress 3-fold to a level similar of the High PFF stimulus, but without affecting streaming potentials or chemotransport. NO and PGE(2) production were stimulated by Low, Medium, and High PFF in a dose-dependent manner. Application of Low PFF using dextran-supplemented medium, enhanced both the NO and PGE(2) response by 3-fold, to a level mimicking the response to High PFF at normal viscosity. These results show that the production of NO and PGE(2) by bone cells can be enhanced in a dose-dependent manner by fluid flow of increasing wall shear stress. Therefore, the stimulus leading to NO and PGE(2) production is the flow-derived shear stress, and not streaming potentials or chemotransport. PMID:11311708

Bakker, A D; Soejima, K; Klein-Nulend, J; Burger, E H

2001-05-01

322

Separation of Spleen Colony Forming Units (CFU-S) from Mouse Bone Marrow Cells Using Velocity Sedimentation in an Isokinetic Gradient of Ficoll in Tissue Culture Medium  

PubMed Central

Spleen colony forming cells (CFU-S) from mouse bone marrow were concentrated using velocity sedimentation in an isokinetic gradient of Ficoll (polysucrose) in tissue culture medium. Following separation, CFU-S were found as a sharp modal population of cells which was discrete from the modal populations of lymphocytes and other cell types. Despite the wide separation of lymphocytes, granulocytes and CFU-S in the density gradient, there was no appreciable resolution of CFU-S specifically committed to erythroid, granulocytic or undifferentiated colony formation. This suggests that committed stem cells either share identical physical properties or are not detected by the spleen colony assay. In the purest fractions from the isokinetic gradient, CFU-S were 2.8- to 4.7-fold purified when compared with mouse bone marrow before separation. ImagesFig 1Fig 2Fig 3

Pretlow, Thomas G.; Williams, Edwin E.; Davis, Maxie L.; Zettergren, Judy G.

1973-01-01

323

Expression pattern of receptor activator of NF?B (RANK) in a series of primary solid tumors and related bone metastases.  

PubMed

Receptor activator of NF?B ligand (RANKL), RANK, and osteoprotegerin (OPG) represent the key regulators of bone metabolism both in normal and pathological conditions, including bone metastases. To our knowledge, no previous studies investigated and compared RANK expression in primary tumors and in bone metastases from the same patient. We retrospectively examined RANK expression by immunohistochemistry in 74 bone metastases tissues from solid tumors, mostly breast, colorectal, renal, lung, and prostate cancer. For 40 cases, tissue from the corresponding primary tumor was also analyzed. Sixty-six (89%) of the 74 bone metastases were RANK-positive and, among these, 40 (59.5%) showed more than 50% of positive tumor cells. The median percentage of RANK-positive cells was 60% in primary tumors and metastases, without any statistically significant difference between the two groups (P=0.194). The same percentage was obtained by considering only cases with availability of samples both from primary and metastasis. Our study shows that RANK is expressed by solid tumors, with high concordance between bone metastasis and corresponding primary tumor. These data highlight the central role of RANK/RANKL/OPG pathway as potential therapeutic target not only in bone metastasis management, but also in the adjuvant setting. PMID:20857484

Santini, Daniele; Perrone, Giuseppe; Roato, Ilaria; Godio, Laura; Pantano, Francesco; Grasso, Donatella; Russo, Antonio; Vincenzi, Bruno; Fratto, Maria Elisabetta; Sabbatini, Roberto; Della Pepa, Chiara; Porta, Camillo; Del Conte, Alessandro; Schiavon, Gaia; Berruti, Alfredo; Tomasino, Rosa Maria; Papotti, Mauro; Papapietro, Nicola; Onetti Muda, Andrea; Denaro, Vincenzo; Tonini, Giuseppe

2011-03-01

324

Primary bone marrow diffuse large B-cell lymphoma accompanying cold agglutinin disease: A case report with review of the literature  

PubMed Central

Cold agglutinin disease (CAD) is a well-recognized complication of lymphoproliferative disorders. It has been previously recognized that cases of primary CAD frequently exhibit underlying malignant lymphoma in the bone marrow. Lymphoplasmacytic lymphoma is the most common subtype of malignant lymphoma; however, diffuse large B-cell lymphoma (DLBCL) has also been documented, albeit extremely rare. The current report presents a case of primary bone marrow DLBCL accompanying CAD. A 76-year-old male presented with fever and fatigue. Laboratory tests revealed anemia and elevated bilirubin and cold agglutinins with a titer of 8,192 at 4°C. Bone marrow biopsy demonstrated DLBCL and systemic surveillance failed to detect tumorous lesions or lymphadenopathy. Following R-THP-COP therapy, cold agglutinins titer was markedly decreased (by <4); however, malignant lymphoma relapsed and cold agglutinin levels increased again (4,096). This is the second documented case of primary bone marrow DLBCL accompanying CAD. Previously, malignant lymphoma exclusively involving the bone marrow, namely primary bone marrow lymphoma (PBML), has been recognized as a rare and aggressive subtype. The analyses of the present study revealed that the incidence of hemolytic anemia in primary bone marrow DLBCL may be high compared with conventional DLBCL. Therefore, additional analyses are required to clarify the clinicopathological features of PBML.

YAMASHITA, TOMOKO; ISHIDA, MITSUAKI; MORO, HIROKO; YUMOTO, HIROFUMI; UCHIBAYASHI, SACHIKO; YOSHII, MIYUKI; NAKANISHI, RYOTA; OKUNO, HIROKO; YOSHIDA, TAKASHI; OKUNO, TAKAFUMI; HODOHARA, KEIKO; OKABE, HIDETOSHI

2014-01-01

325

Functional outcome after endoprosthetic limb-salvage therapy of primary bone tumours—a comparative analysis using the MSTS score, the TESS and the RNL index  

Microsoft Academic Search

Limb-saving therapy for primary bone tumours is the treatment of choice. We aimed at analysing the quality of life of this\\u000a group of patients by combining three different tools. Eighty-seven patients (46 females, 41 males) with a primary bone tumour\\u000a of the extremity who had undergone endoprosthetic reconstruction between 1982 and 2000 were included in this retrospective\\u000a study. The median

P. U. Tunn; D. Pomraenke; U. Goerling; P. Hohenberger

2008-01-01

326

Isolation and culture of primary osteocytes from the long bones of skeletally mature and aged mice  

PubMed Central

The purpose of this work was to establish a methodology to enable the isolation and study of osteocytes from skeletally mature young (4-month-old) and old (22-month-old) mice. The location of osteocytes deep within bone is ideal for their function as mechanosensors. However, this location makes the observation and study of osteocytes in vivo technically difficult. Osteocytes were isolated from murine long bones through a process of extended collagenase digestions combined with EDTA-based decalcification. A tissue homogenizer was used to reduce the remaining bone fragments to a suspension of bone particles, which were placed in culture to yield an outgrowth of osteocyte-like cells. All of the cells obtained from this outgrowth that displayed an osteocyte-like morphology stained positive for the osteocyte marker E11/GP38.[Q1] The osteocyte phenotype was further confirmed by a lack of staining for alkaline phosphatase and the absence of collagen1a1 expression. The outgrowth of osteocytes also expressed additional osteocyte-specific genes such as Sost and Mepe. This technique facilitates the isolation of osteocytes from skeletally mature bone. This novel enabling methodology should prove useful in advancing our understanding of the roles mature osteocytes play in bone health and disease.

Stern, Amber Rath; Stern, Matthew M.; Van Dyke, Mark E.; Jahn, Katharina; Prideaux, Matthew; Bonewald, Lynda F.

2013-01-01

327

Resveratrol prevents CsA inhibition of proliferation and osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells through an ER\\/NO\\/cGMP pathway  

Microsoft Academic Search

The purpose of this study was to investigate the in vitro effects of resveratrol (RSVL) and cyclosporin A (CsA) on proliferation and osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cell (BMSC) cultures. Application of RSVL (10?8–10?6moll?1) resulted in a dose-dependent increase in [3H]-thymidine incorporation, alkaline phosphatase (ALP) activity and calcium deposition of BMSCs cultures, which was accompanied with the

Li Hua Song; Wei Pan; Yan Hui Yu; Leigh Darryl Quarles; Hong Hao Zhou; Zhou Sheng Xiao

2006-01-01

328

PARTIAL PURIFICATION AND SOME PROPERTIES OF THE FACTOR IN NORMAL AND LEUKAEMIC HUMAN URINE STIMULATING MOUSE BONE MARROW COLONY GROWTH IN VITRO  

Microsoft Academic Search

The properties of the factor in human urine which stimulates mouse bone marrow colony growth in vitro were examined. The colony stimulating activity of urine was not lost on concentration of urine, exposure to RNA-ase, DNA-ase, ether, 8M urea, pH in the range 2–12, or on fractionation by precipitation with ethanol or ammonium sulphate. Activity was lost on heating (90°\\/30?),

ER Stanley; D Metcalf

1969-01-01

329

Generation of Large Numbers of Dendritic Cells from Mouse Bone Marrow Cultures Supplemented with Granulocyte\\/Macrophage Colony-stimulating Factor  

Microsoft Academic Search

Summary Antigen-presenting, major histocompatibility complex (MHC) class II-rich dendritic cells are known to arise from bone marrow. However, marrow lacks mature dendritic cells, and substantial numbers of proliferating less-mature cells have yet to be identified. The methodology for inducing dendritic cell growth that was recently described for mouse blood now has been modified to MHC class II-negative precursors in marrow.

Kayo Inaba; Muneo Inaba; Nikolaus Romani; Hideki Aya; Masashi Deguchi; Susumu Ikehara; Shigeru Muramatsu; Ralph M. Steinmanll

1992-01-01

330

Embryonic stem cell therapy improves bone quality in a model of impaired fracture healing in the mouse; tracked temporally using in vivo micro-CT.  

PubMed

In the current study, we used an estrogen-deficient mouse model of osteoporosis to test the efficacy of a cell-generated bone tissue construct for bone augmentation of an impaired healing fracture. A reduction in new bone formation at the defect site was observed in ovariectomized fractures compared to the control group using repeated measures in vivo micro-computed tomography (?CT) imaging over 4weeks. A significant increase in the bone mineral density (BMD), trabecular bone volume ratio, and trabecular number, thickness and connectivity were associated with fracture repair in the control group, whereas the fractured bones of the ovariectomized mice exhibited a loss in all of these parameters (p<0.001). In a separate group, ovariectomized fractures were treated with murine embryonic stem (ES) cell-derived osteoblasts loaded in a three-dimensional collagen I gel and recovery of the bone at the defect site was observed. A significant increase in the trabecular bone volume ratio (p<0.001) and trabecular number (p<0.01) was observed by 4weeks in the fractures treated with cell-loaded collagen matrix compared to those treated with collagen I alone. The stem cell-derived osteoblasts were identified at the fracture site at 4weeks post-implantation through in situ hybridization histochemistry. Although this cell tracking method was effective, the formation of an ectopic cellular nodule adjacent to the knee joints of two mice suggested that alternative in vivo cell tracking methods should be employed in order to definitively assess migration of the implanted cells. To our knowledge, this study is the first of its kind to examine the efficacy of stem cell therapy for fracture repair in an osteoporosis-related fracture model in vivo. The findings presented provide novel insight into the use of stem cell therapies for bone injuries. PMID:24780879

Taiani, J T; Buie, H R; Campbell, G M; Manske, S L; Krawetz, R J; Rancourt, D E; Boyd, S K; Matyas, J R

2014-07-01

331

Detection of rodent liver carcinogen genotoxicity by the alkaline single-cell gel electrophoresis (Comet) assay in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow).  

PubMed

We have recently designed a simple method for applying the alkaline single-cell gel electrophoresis (SCG) assay to mouse organs. With this method, each organ is minced, suspended in chilled homogenizing buffer containing NaCl and Na2EDTA, gently homogenized using a Potter-type homogenizer set in ice, and then centrifuged nuclei are used for the alkaline SCG assay. In the present study, we used the method to assess the genotoxicity of 8 rodent hepatic carcinogens in 5 mouse organs (liver, lung, kidney, spleen, and bone marrow). The carcinogens we studied were p-aminoazobenzene, auramine, 2,4-diaminotoluene, p-dichlorobenzene, ethylene thiourea (ETU), styrene-7,8-oxide, phenobarbital sodium, and benzene-1,2,3,4,5,6-hexachloride (BHC); except for p-aminoazobenzene, they do not induce micronuclei in mouse bone marrow cells. Mice were sacrificed 3 and 24 h after the administration of each carcinogen. p-Aminoazobenzene, ETU, and styrene-7,8-oxide induced alkaline labile DNA lesions in all of the organs studied. Auramine, 2,4-diaminotoluene, p-dichlorobenzene, and phenobarbital sodium also produced lesions, but their effect was greatest in the liver. BHC, which is not genotoxic in in vitro tests, did not show any effects. We suggest that it may be possible to use the alkaline SCG assay to detect in vivo activity of chemicals whose genotoxicity is not expressed in bone marrow cells. PMID:9268046

Sasaki, Y F; Izumiyama, F; Nishidate, E; Matsusaka, N; Tsuda, S

1997-07-14

332

Neurotrophism of bone marrow stromal cells to embryonic stem cells: noncontact induction and transplantation to a mouse ischemic stroke model.  

PubMed

Embryonic stem (ES) cell-derived cell products may serve as a source of cells for regenerative medicine. Currently available technologies for the induction of ES cells into neural lineage cells require extended culturing in vitro and complex procedural manipulations, with variable yields of heterogeneous cells, which have hindered the prospective use of cell derivatives for treatment of ischemic stroke. We established a simple and efficient method to derive mouse ES cells into neural lineage cells using an 8-day coculture with the bone marrow stromal cells MS5, followed by a 6-day propagation culture and a 4-day selection culture. The protocol generated a relatively high yield of neural lineage cells without any mesodermal and endodermal lineage commitment. In in vivo study, these derived cells could improve the cognitive function of ischemic stroke mice. Three weeks after transplantation, migration of implanted cells to lesioned areas was noted. It was also evident of a normalization of pyramidal neuron density and morphology in hippocampal CA1 region. One (1/17) episode of teratoma development was noted. Data suggested that MS5 cells may exert a neurotrophic effect to enhance neural differentiation of ES cells and MS5-induced ES cell-derived cells appeared to be applicable to cell therapy for ischemic stroke. PMID:19622227

Yang, Tao; Tsang, Kam Sze; Poon, Wai Sang; Ng, Ho Keung

2009-01-01

333

Effect of bone marrow transplantation on enzyme levels and clinical course in the neurologically affected twitcher mouse.  

PubMed Central

The effect of allogeneic bone marrow transplantation (BMT) was investigated in the neurologically affected twitcher mouse, a model for human Krabbe's disease. Twitcher mice have a hereditary deficiency of the lysosomal enzyme galactosylceramidase, which causes growth delay, tremor, and paralysis of the hind legs. Death occurs at 30-40 d of age. After BMT galactosylceramidase activity increased to donor levels in hemopoietic organs. In lung, heart, and liver, galactosylceramidase activity rose to levels intermediate between those of twitcher and normal mice. Increased galactosylceramidase activity in liver parenchymal cells indicated uptake of the donor enzyme by recipient cells of nonhemopoietic origin. Enzyme activity also increased in kidney tissue. BMT resulted in a gradual increase in galactosylceramidase activity in the central nervous system to 15% of normal donor levels. A 5-6-fold increase in galactosylceramidase activity was found in the peripheral nervous system. This increase in enzyme activity was accompanied by a partial alleviation of neurological symptoms. In particular, paralysis of the hind legs was prevented by BMT. BMT led to a modest restoration of growth and prolonged survival. In several cases, the mice survived for more than 100 d, but eventually all animals died with severe neurological disease.

Hoogerbrugge, P M; Poorthuis, B J; Romme, A E; van de Kamp, J J; Wagemaker, G; van Bekkum, D W

1988-01-01

334

Cell Fusion Reprogramming Leads to a Specific Hepatic Expression Pattern during Mouse Bone Marrow Derived Hepatocyte Formation In Vivo  

PubMed Central

The fusion of bone marrow (BM) hematopoietic cells with hepatocytes to generate BM derived hepatocytes (BMDH) is a natural process, which is enhanced in damaged tissues. However, the reprogramming needed to generate BMDH and the identity of the resultant cells is essentially unknown. In a mouse model of chronic liver damage, here we identify a modification in the chromatin structure of the hematopoietic nucleus during BMDH formation, accompanied by the loss of the key hematopoietic transcription factor PU.1/Sfpi1 (SFFV proviral integration 1) and gain of the key hepatic transcriptional regulator HNF-1A homeobox A (HNF-1A/Hnf1a). Through genome-wide expression analysis of laser captured BMDH, a differential gene expression pattern was detected and the chromatin changes observed were confirmed at the level of chromatin regulator genes. Similarly, Tranforming Growth Factor-?1 (TGF-?1) and neurotransmitter (e.g. Prostaglandin E Receptor 4 [Ptger4]) pathway genes were over-expressed. In summary, in vivo BMDH generation is a process in which the hematopoietic cell nucleus changes its identity and acquires hepatic features. These BMDHs have their own cell identity characterized by an expression pattern different from hematopoietic cells or hepatocytes. The role of these BMDHs in the liver requires further investigation.

Arza, Elvira; Alvarez-Barrientos, Alberto; Fabregat, Isabel; Garcia-Bravo, Maria; Meza, Nestor W.; Segovia, Jose C.

2012-01-01

335

STAT3 mutations identified in human hematologic neoplasms induce myeloid malignancies in a mouse bone marrow transplantation model  

PubMed Central

STAT3 protein phosphorylation is a frequent event in various hematologic malignancies and solid tumors. Acquired STAT3 mutations have been recently identified in 40% of patients with T-cell large granular lymphocytic leukemia, a rare T-cell disorder. In this study, we investigated the mutational status of STAT3 in a large series of patients with lymphoid and myeloid diseases. STAT3 mutations were identified in 1.6% (4 of 258) of patients with T-cell neoplasms, in 2.5% (2 of 79) of patients with diffuse large B-cell lymphoma but in no other B-cell lymphoma patients (0 of 104) or patients with myeloid malignancies (0 of 96). Functional in vitro assays indicated that the STAT3Y640F mutation leads to a constitutive phosphorylation of the protein. STA21, a STAT3 small molecule inhibitor, inhibited the proliferation of two distinct STAT3 mutated cell lines. Using a mouse bone marrow transplantation assay, we observed that STAT3Y640F expression leads to the development of myeloproliferative neoplasms with expansion of either myeloid cells or megakaryocytes. Together, these data indicate that the STAT3Y640F mutation leads to constitutive activation of STAT3, induces malignant hematopoiesis in vivo, and may represent a novel therapeutic target in some lymphoid malignancies.

Couronne, Lucile; Scourzic, Laurianne; Pilati, Camilla; Valle, Veronique Della; Duffourd, Yannis; Solary, Eric; Vainchenker, William; Merlio, Jean-Philippe; Beylot-Barry, Marie; Damm, Frederik; Stern, Marc-Henri; Gaulard, Philippe; Lamant, Laurence; Delabesse, Eric; Merle-Beral, Helene; Nguyen-Khac, Florence; Fontenay, Michaela; Tilly, Herve; Bastard, Christian; Zucman-Rossi, Jessica; Bernard, Olivier A.; Mercher, Thomas

2013-01-01

336

In Vitro Assays Using Primary Embryonic Mouse Lymphatic Endothelial Cells Uncover Key Roles for FGFR1 Signalling in Lymphangiogenesis  

PubMed Central

Despite the importance of blood vessels and lymphatic vessels during development and disease, the signalling pathways underpinning vessel construction remain poorly characterised. Primary mouse endothelial cells have traditionally proven difficult to culture and as a consequence, few assays have been developed to dissect gene function and signal transduction pathways in these cells ex vivo. Having established methodology for the purification, short-term culture and transfection of primary blood (BEC) and lymphatic (LEC) vascular endothelial cells isolated from embryonic mouse skin, we sought to optimise robust assays able to measure embryonic LEC proliferation, migration and three-dimensional tube forming ability in vitro. In the course of developing these assays using the pro-lymphangiogenic growth factors FGF2 and VEGF-C, we identified previously unrecognised roles for FGFR1 signalling in lymphangiogenesis. The small molecule FGF receptor tyrosine kinase inhibitor SU5402, but not inhibitors of VEGFR-2 (SU5416) or VEGFR-3 (MAZ51), inhibited FGF2 mediated LEC proliferation, demonstrating that FGF2 promotes proliferation directly via FGF receptors and independently of VEGF receptors in primary embryonic LEC. Further investigation revealed that FGFR1 was by far the predominant FGF receptor expressed by primary embryonic LEC and correspondingly, siRNA-mediated FGFR1 knockdown abrogated FGF2 mediated LEC proliferation. While FGF2 potently promoted LEC proliferation and migration, three dimensional tube formation assays revealed that VEGF-C primarily promoted LEC sprouting and elongation, illustrating that FGF2 and VEGF-C play distinct, cooperative roles in lymphatic vascular morphogenesis. These assays therefore provide useful tools able to dissect gene function in cellular events important for lymphangiogenesis and implicate FGFR1 as a key player in developmental lymphangiogenesis in vivo.

Betterman, Kelly L.; Harvey, Natasha L.

2012-01-01

337

Quantitative trait loci, genes, and polymorphisms that regulate bone mineral density in mouse  

PubMed Central

This is an in silico analysis of data available from genome-wide scans. Through analysis of QTL, genes and polymorphisms that regulate BMD, we identified 82 BMD QTL, 191 BMD-associated (BMDA) genes, and 83 genes containing known BMD-associated polymorphisms (BMDAP). The catalogue of all BMDA/BMDAP genes and relevant literatures are provided. In total, there are substantially more BMDA/BMDAP genes in regions of the genome where QTL have been identified than in non-QTL regions. Among 191 BMDA genes and 83 BMDAP genes, 133 and 58 are localized in QTL region, respectively. The difference was still noticeable for the chromosome distribution of these genes between QTL and non-QTL regions. These results have allowed us to generate an integrative profile of QTL, genes, polymorphisms that determine BMD. These data could facilitate more rapid and comprehensive identification of causal genes underlying the determination of BMD in mouse and provide new insights into how BMD is regulated in humans.

Xiong, Qing; Jiao, Yan; Hasty, Karen A.; Canale, S. Terry; Stuart, John M.; Beamer, Wesley G.; Deng, Hong-Wen; Baylink, David; Gu, Weikuan

2010-01-01

338

WNT-3A and WNT-5A counteract lipopolysaccharide-induced pro-inflammatory changes in mouse primary microglia.  

PubMed

Surveying microglia, the resident macrophage-like cells in the central nervous system, continuously screen their surroundings to sense imbalance in tissue homeostasis. Their activity is tightly regulated in both a pro- and anti-inflammatory manner. We have previously shown that the lipoglycoproteins WNT-3A and WNT-5A drive pro-inflammatory transformation in primary mouse microglia cells, arguing that WNTs have a role in the modulation of the central nervous system immune response. In this study, we address the effects of recombinant WNT-3A and WNT-5A on lipopolysaccharide (LPS)-activated mouse primary microglia to investigate the putative anti-inflammatory modulation of microglia by WNTs. While both WNT-3A and WNT-5A alone induce an up-regulation of cyclooxygenase 2 (COX2), a generic pro-inflammatory microglia marker, LPS exceeds these effects dramatically. However, combination of LPS and WNTs results in a dose-dependent decrease in LPS-induced cyclooxygenase 2 protein and mRNA expression. In conclusion, our data suggest that WNTs have a dual and context-dependent effect on microglia acting in a homeostatic pro- and anti-inflammatory manner. PMID:23534675

Halleskog, Carina; Schulte, Gunnar

2013-06-01

339

Graft versus Host Disease in the Bone Marrow, Liver and Thymus Humanized Mouse Model  

PubMed Central

Mice bearing a “humanized” immune system are valuable tools to experimentally manipulate human cells in vivo and facilitate disease models not normally possible in laboratory animals. Here we describe a form of GVHD that develops in NOD/SCID mice reconstituted with human fetal bone marrow, liver and thymus (NS BLT mice). The skin, lungs, gastrointestinal tract and parotid glands are affected with progressive inflammation and sclerosis. Although all mice showed involvement of at least one organ site, the incidence of overt clinical disease was approximately 35% by 22 weeks after reconstitution. The use of hosts lacking the IL2 common gamma chain (NOD/SCID/?c?/?) delayed the onset of disease, but ultimately did not affect incidence. Genetic analysis revealed that particular donor HLA class I alleles influenced the risk for the development of GVHD. At a cellular level, GVHD is associated with the infiltration of human CD4+ T cells into the skin and a shift towards Th1 cytokine production. GVHD also induced a mixed M1/M2 polarization phenotype in a dermal murine CD11b+, MHC class II+ macrophage population. The presence of xenogenic GVHD in BLT mice both presents a major obstacle in the use of humanized mice and an opportunity to conduct preclinical studies on GVHD in a humanized model.

Greenblatt, Matthew B.; Vbranac, Vladimir; Tivey, Trevor; Tsang, Kelly; Tager, Andrew M.; Aliprantis, Antonios O.

2012-01-01

340

The genotoxic and cytotoxic effects of nimesulide in the mouse bone marrow.  

PubMed

Abstract Genotoxicity of nimesulide (NM) was evaluated by employing bone marrow (BM) chromosomal aberration (CA) and micronucleus assays in Swiss albino mice. For BM CA assay, mice of either sex were treated orally with 1.5, 2.5 and 5?mg body weight solution of NM in 0.2?mL of 0.05% CMC (carboxy methyl cellulose) daily for 4, 13, 28 and 40 weeks. Treatment induced dose-dependent and significantly depressed mitotic activity and increase in CAs per cell in the BM cells after 13 weeks of treatment at all dose levels. In micronucleus assay, male mice were treated orally with the same dose levels and sampling durations as for CA assay. Treatment increased the percentage of micronucleated polychromatic erythrocytes frequency and showed a statistically significant reduction in polychromatic erythrocyte/normochromatic erythrocyte ratio, as compared to control groups. Cyclophosphamide (40?mg/kg) was used as clastogen (positive control) and yielded the expected positive results. Cytotoxicity was observed in the 8-week recovery period after 40 weeks of dosing, but it was not significant. On the basis of these findings, it may be concluded that in the long term, NM, or its biotransformed product, is genotoxic and cytotoxic for BM cells of mice in vivo. PMID:24164450

Tripathi, Rina; Tripathi, Pankaj; Pancholi, Shyam S; Patel, Chhagan N

2014-07-01

341

Primary malignant bone tumors and solitary metastases of the thoracolumbar spine: results by management with total en bloc spondylectomy  

PubMed Central

Primary malignant spinal tumors and solitary vertebral metastases of selected tumor entities in the thoracolumbar spine are indications for total en bloc spondylectomy (TES). This study aimed to describe our oncological and surgical management and to analyze the treatment results by management with TES for extra- and intracompartmental solitary spinal metastases and primary malignant vertebral bone tumors. In 15 patients (3 malignant bone tumors and 12 solitary metastases), tumors were distributed in the thoracic (n = 8) and lumbar (n = 7) spine. Tumors were classified as intra- (n = 8) and extracompartmental (n = 7). All patients underwent TES via a laterally extended posterior approach followed by dorsoventral reconstruction. Function and quality of life were assessed by Oswestry disability index (ODI) and SF-36 score. At follow-up (100%; mean: 33 ± 22 months), 11 patients had no evidence of disease. Two patients were alive with the disease and two were dead of the disease (no primary bone tumors). Histology revealed negative margins (R0) in all patients with wide (n = 11) and marginal (n = 4) resections. Two patients developed pulmonal metastases of which they died at 4 and 16 months of survival. No local recurrence was observed. Major complications did not occur. TES resulted in an acceptable outcome in the quality of life and function. TES is a demanding procedure reaching wide to marginal resections in a curative approach. In conjunction with multimodal therapies, local recurrences can effectively be prevented while control of distant disease needs to be improved. Proper selection of adequate patients combined with careful surgical planning are prerequisites for low complication rates, acceptable function and improved overall prognosis.

Melcher, Ingo; Khodadadyan-Klostermann, Cyrus; Tohtz, Stefan; Smolny, Mirko; Stockle, Ulrich; Haas, Norbert P.; Schaser, Klaus-Dieter

2007-01-01

342

THE ENHANCING EFFECT OF BONE MARROW CELLS ON THE PRIMARY IMMUNE RESPONSE OF THE ISOLATED PERFUSED SPLEEN  

PubMed Central

The addition of bone marrow cells or peripheral lymphocytes to the isolated pig spleen markedly enhanced the primary antibody response after 3-day perfusion and antigenic challenge in vitro. The splenic preparation without added cells or with the addition of marrow cells to an irradiated spleen gave a limited response. Contributory evidence is provided that at least two distinct cell types are needed for antibody production. For optimal antibody response by an isolated perfused spleen, marrow cells or peripheral lymphocytes should be added to the system.

Atkins, Robert C.; Robinson, William A.; Eiseman, Ben

1970-01-01

343

Diabetes impairs mobilization of mouse bone marrow-derived Lin(-)/VEGF-R2(+) progenitor cells.  

PubMed

Endothelial progenitor cells circulating in the peripheral blood (PB) contribute to vascular repair. This study aimed to evaluate the potential of a 'cocktail' consisting of erythropoietin, granulocyte colony-stimulating factor and tetrahydrobiopterin to mobilize hematopoietic lineage negative/vascular endothelial growth factor receptor 2 positive (Lin(-)/VEGF-R2(+)) cells from the bone marrow (BM) to PB in non-diabetic and diabetic mice. Diabetes was induced in mice by intraperitoneal injection of streptozotocin. Diabetic mice were studied after 16weeks of hyperglycemia. Half the mice in each group (non-diabetic and diabetic) received daily intraperitoneal injections of the cocktail for 6 consecutive days while the other half received vehicle buffer. Mobilization of Lin(-)/VEGF-R2(+) cells, which were expanded in MCP301 medium, was evaluated after isolating them from BM and PB and their phenotypic and morphological properties were studied. We found that 16weeks of diabetes affected neither the total number of BM mononucleated cells nor the number of Lin(-)/VEGF-R2(+) cells in BM compared with non-diabetic controls. In non-diabetic mice, cocktail treatment resulted in a significant decrease in BM Lin(-)/VEGF-R2(+) cells, paralleled by a significant increase of these cells in PB. Such changes in the number of Lin(-)/VEGF-R2(+) cells in BM and PB after the cocktail treatment were less marked in diabetic mice. In vitro studies of BM Lin(-)/VEGF-R2(+) cells from diabetic and non-diabetic mice did not reveal any differences in either phenotypes or colony forming potential. These findings indicate that diabetes impairs the mobilization of Lin(-)/VEGF-R2(+) cells from BM to PB. Impaired mobilization of BM Lin(-)/VEGF-R2(+) cells soon after the onset of diabetes may contribute to complications such as diabetic retinopathy. PMID:23714230

Barthelmes, D; Irhimeh, M R; Gillies, M C; Karimipour, M; Zhou, M; Zhu, L; Shen, W Y

2013-10-01

344

Mechanical stimulation and intermittent parathyroid hormone treatment induce disproportional osteogenic, geometric, and biomechanical effects in growing mouse bone  

PubMed Central

Mechanical loading and intermittent parathyroid (iPTH) treatment are both osteoanabolic stimuli, and are regulated by partially overlapping cellular signaling pathways. iPTH has been shown clinically to be effective in increasing bone mass and reducing fracture risk. Likewise, mechanical stimulation can significantly enhance bone apposition and prevent bone loss, but its clinical effects on fracture susceptibility are less certain. Many of the osteogenic effects of iPTH are localized to biomechanically suboptimal bone surfaces, whereas mechanical loading directs new bone formation to high-stress areas and not to strain-neutral areas. These differences in localization in new tissue, resulting from load-induced vs iPTH-induced bone accumulation, should affect the relation between bone mass and bone strength, or “tissue economy.” We investigated the changes in bone mass and strength induced by 6 wks mechanical loading, and compared them to changes induced by 6 wks iPTH treatment. Loading and iPTH both increased ulnar bone accrual, as measured by bone mineral density and content, and fluorochrome-derived bone formation. iPTH induced a significantly greater increase in bone mass than loading, but ulnar bone strength was increased approximately the same amount by both treatments. Mechanical loading during growth can spatially optimize new bone formation to improve structural integrity with a minimal increase in mass, thereby increasing tissue economy i.e., the amount of strength returned per unit bone mass added. Furthermore, exercise studies in which only small changes in bone mass are detected might be more beneficial to bone health and fracture resistance than has commonly been presumed.

McAteer, Maureen E.; Niziolek, Paul J.; Ellis, Shana N.; Alge, Daniel L.; Robling, Alexander G.

2011-01-01

345

VCP associated inclusion body myopathy and paget disease of bone knock-in mouse model exhibits tissue pathology typical of human disease.  

PubMed

Dominant mutations in the valosin containing protein (VCP) gene cause inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD). We have generated a knock-in mouse model with the common R155H mutation. Mice demonstrate progressive muscle weakness starting approximately at the age of 6 months. Histology of mutant muscle showed progressive vacuolization of myofibrils and centrally located nuclei, and immunostaining shows progressive cytoplasmic accumulation of TDP-43 and ubiquitin-positive inclusion bodies in quadriceps myofibrils and brain. Increased LC3-II staining of muscle sections representing increased number of autophagosomes suggested impaired autophagy. Increased apoptosis was demonstrated by elevated caspase-3 activity and increased TUNEL-positive nuclei. X-ray microtomography (uCT) images show radiolucency of distal femurs and proximal tibiae in knock-in mice and uCT morphometrics shows decreased trabecular pattern and increased cortical wall thickness. Bone histology and bone marrow derived macrophage cultures in these mice revealed increased osteoclastogenesis observed by TRAP staining suggestive of Paget bone disease. The VCP(R155H/+) knock-in mice replicate the muscle, bone and brain pathology of inclusion body myopathy, thus representing a useful model for preclinical studies. PMID:20957154

Badadani, Mallikarjun; Nalbandian, Angèle; Watts, Giles D; Vesa, Jouni; Kitazawa, Masashi; Su, Hailing; Tanaja, Jasmin; Dec, Eric; Wallace, Douglas C; Mukherjee, Jogeshwar; Caiozzo, Vincent; Warman, Matthew; Kimonis, Virginia E

2010-01-01

346

BMP-Non-Responsive Sca1+CD73+CD44+ Mouse Bone Marrow Derived Osteoprogenitor Cells Respond to Combination of VEGF and BMP-6 to Display Enhanced Osteoblastic Differentiation and Ectopic Bone Formation  

PubMed Central

Clinical trials on fracture repair have challenged the effectiveness of bone morphogenetic proteins (BMPs) but suggest that delivery of mesenchymal stem cells (MSCs) might be beneficial. It has also been reported that BMPs could not increase mineralization in several MSCs populations, which adds ambiguity to the use of BMPs. However, an exogenous supply of MSCs combined with vascular endothelial growth factor (VEGF) and BMPs is reported to synergistically enhance fracture repair in animal models. To elucidate the mechanism of this synergy, we investigated the osteoblastic differentiation of cloned mouse bone marrow derived MSCs (D1 cells) in vitro in response to human recombinant proteins of VEGF, BMPs (-2, -4, -6, -9) and the combination of VEGF with BMP-6 (most potent BMP). We further investigated ectopic bone formation induced by MSCs pre-conditioned with VEGF, BMP-6 or both. No significant increase in mineralization, phosphorylation of Smads 1/5/8 and expression of the ALP, COL1A1 and osterix genes was observed upon addition of VEGF or BMPs alone to the cells in culture. The lack of CD105, Alk1 and Alk6 expression in D1 cells correlated with poor response to BMPs indicating that a greater care in the selection of MSCs is necessary. Interestingly, the combination of VEGF and BMP-6 significantly increased the expression of ALP, COL1A1 and osterix genes and D1 cells pre-conditioned with VEGF and BMP-6 induced greater bone formation in vivo than the non-conditioned control cells or the cells pre-conditioned with either VEGF or BMP-6 alone. This enhanced bone formation by MSCs correlated with higher CADM1 expression and OPG/RANKL ratio in the implants. Thus, combined action of VEGF and BMP on MSCs enhances osteoblastic differentiation of MSCs and increases their bone forming ability, which cannot be achieved through use of BMPs alone. This strategy can be effectively used for bone repair.

Madhu, Vedavathi; Li, Ching-Ju; Dighe, Abhijit S.; Balian, Gary; Cui, Quanjun

2014-01-01

347

Comparison of differentiation potential of male mouse adipose tissue and bone marrow derived-mesenchymal stem cells into germ cells  

PubMed Central

Background: Recent publications about differentiation of stem cells to germ cells have motivated researchers to make new approaches to infertility. In vitro production of germ cells improves understanding differentiation process of male and female germ cells. Due to the problem of using embryonic stem cells (ESC), it’s necessary the mentioned cells be replaced with some adult multi-potent stem cells in laboratories. Objective: The aim of this study was to obtain germ cells from appropriate source beyond ESC and compare differential potentials of adipocytes derived stem cells (ADMSCs) with bone marrow derived stem cells (BMMSCs). Materials and Methods: To find multi-potential entity, after providing purified ADMSCs and BMMSCs, differentiation to osteoblast and adipocyte was confirmed by using appropriate culture medium. To confirm mesenchymal lineage production superficial markers (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flowcytometry. Then the cells were differentiated to germ cells in inductive medium containing retinoic acid for 7days. To evaluate germ cells characteristic markers [Dazl (Deleted in azoospermia-like), Mvh (Mouse vasa homolog gene), Stra8 (Stimulated by retinoic acid) and Scp3 (Synaptonemal complex protein 3)] flowcytometry, imunoflorescence and real time PCR were used. Results: Both types of cells were able to differentiate into osteoblast and adipocyte cells and presentation of stem cell superficial markers (CD90, CD44) and absence of endothelial and blood cell markers (CD31, CD45) were confirmative The flowcytometry, imunoflorescence and real time PCR results showed remarkable expression of germ cells characteristic markers (Mvh, Dazl, Stra8, and Scp3). Conclusion: It was found that although ADMSCs were attained easier and also cultured and differentiated rapidly, germ cell markers were expressed in BMMSCs significantly more than ADMSCs. This article extracted from M.Sc. thesis. (Maryam Hosseinzadeh Shirzeily)

Hosseinzadeh Shirzeily, Maryam; Pasbakhsh, Parichehr; Amidi, Fardin; Mehrannia, Kobra; Sobhani, Aligholi

2013-01-01

348

Initial Binding and Recellularization of Decellularized Mouse Lung Scaffolds with Bone Marrow-Derived Mesenchymal Stromal Cells  

PubMed Central

Recellularization of whole decellularized lung scaffolds provides a novel approach for generating functional lung tissue ex vivo for subsequent clinical transplantation. To explore the potential utility of stem and progenitor cells in this model, we investigated recellularization of decellularized whole mouse lungs after intratracheal inoculation of bone marrow-derived mesenchymal stromal cells (MSCs). The decellularized lungs maintained structural features of native lungs, including intact vasculature, ability to undergo ventilation, and an extracellular matrix (ECM) scaffold consisting primarily of collagens I and IV, laminin, and fibronectin. However, even in the absence of intact cells or nuclei, a number of cell-associated (non-ECM) proteins were detected using mass spectroscopy, western blots, and immunohistochemistry. MSCs initially homed and engrafted to regions enriched in types I and IV collagen, laminin, and fibronectin, and subsequently proliferated and migrated toward regions enriched in types I and IV collagen and laminin but not provisional matrix (fibronectin). MSCs cultured for up to 1 month in either basal MSC medium or in a small airways growth media (SAGM) localized in both parenchymal and airway regions and demonstrated several different morphologies. However, while MSCs cultured in basal medium increased in number, MSCs cultured in SAGM decreased in number over 1 month. Under both media conditions, the MSCs predominantly expressed genes consistent with mesenchymal and osteoblast phenotype. Despite a transient expression of the lung precursor TTF-1, no other airway or alveolar genes or vascular genes were expressed. These studies highlight the power of whole decellularized lung scaffolds to study functional recellularization with MSCs and other cells.

Daly, Amanda B.; Wallis, John M.; Borg, Zachary D.; Bonvillain, Ryan W.; Deng, Bin; Ballif, Bryan A.; Jaworski, Diane M.; Allen, Gilman B.

2012-01-01

349

Inhibition of DPP4/CD26 and dmPGE? treatment enhances engraftment of mouse bone marrow hematopoietic stem cells.  

PubMed

Enhancing the engraftment of hematopoietic stem cells (HSC) is especially important when times to engraftment are prolonged due either to limiting numbers of HSC in the donor graft or to intrinsic slower engrafting time of the tissue sources of HSC. Both inhibition of dipeptidylpeptidase (DPP) 4/CD26 and treatment of cells with 16,16 dimethyl prostaglandin E2 (dmPGE2) have been shown to enhance hematopoietic stem cell engraftment in murine transplantation models and have been evaluated in clinical settings for their influence on engraftment of cord blood cells, a tissue source of HSC known to manifest an extended time to engraftment of donor cells compared to that of bone marrow (BM) and mobilized peripheral blood for hematopoietic cell transplantation (HCT). Herein, we present new experimental data, using a CD45(+) head-to-head congenic model of donor mouse BM cells for engraftment of lethally irradiated mice, demonstrating that similar levels of enhanced engraftment are detected by pulsing donor BM cells with diprotin A, a DPP4 inhibitor, or with dmPGE2 prior to infusion, or by pretreating recipient mice with sitagliptin, also a DPP4 inhibitor, by oral gavage. Moreover, the combined effects of pretreating the donor BM cells with dmPGE2 in context of pretreating the recipient mice with sitagliptin after the administration of a lethal dose of radiation resulted in significantly enhanced competitively repopulating HCT compared to either treatment alone. This information is highly relevant to the goal of enhancing engraftment in human clinical HCT. PMID:24602918

Broxmeyer, Hal E; Pelus, Louis M

2014-01-01

350

Effects of heavy ion to the primary culture of mouse brain cells  

NASA Technical Reports Server (NTRS)

To investigate effects of low dose heavy particle radiation to CNS system, we adopted mouse neonatal brain cells in culture being exposed to heavy ions by HIMAC at NIRS and NSRL at BNL. The applied dose varied from 0.05 Gy up to 2.0 Gy. The subsequent biological effects were evaluated by an induction of apoptosis and neuron survival focusing on the dependencies of the animal strains, SCID, B6, B6C3F1, C3H, used for brain cell culture, SCID was the most sensitive and C3H the least sensitive to particle radiation as evaluated by 10% apoptotic criterion. The LET dependency was compared with using SCID and B6 cells exposing to different ions (H, C, Ne, Si, Ar, and Fe). Although no detectable LET dependency was observed in the high LET (55-200 keV/micrometers) and low dose (<0.5 Gy) regions. The survivability profiles of the neurons were different in the mouse strains and ions. In this report, a result of memory and learning function to adult mice after whole-body and brain local irradiation at carbon ion and iron ion.

Nojima, Kumie; Nakadai, Taeko; Kohno, Yukio; Vazquez, Marcelo E.; Yasuda, Nakahiro; Nagaoka, Shunji

2004-01-01

351

Correlation between serum levels of free light chain and phenotype of plasma cells in bone marrow in primary AL amyloidosis.  

PubMed

To investigate whether there is a correlation between subtypes of plasma cells in the bone marrow and the production of M-protein, flow cytometry and serum free light chain (FLC) analyses were carried out in 17 patients with primary systemic AL amyloidosis (mean age, 59.9+/-8.8 years) and controls with M-protein (MGUS controls, n=6) and without it (negative controls, n=9). The patients showed a significantly higher value in the serum predominant FLC:serum creatinine ratio (43.8+/-63.2) and CD38++ CD19- CD56+ subpopulation (monoclonal plasma cells) (2.57+/-5.35%) than either the negative (p<0.0005 and p<0.001, respectively) or MGUS controls (p<0.05). With respect to maturation of plasma cells in the bone marrow, the intermediate (MPC-1+ CD45- CD49e-) and mature (MPC-1+ CD45+ CD49e-) subtypes were significantly higher (49.2+/-23.2%, p<0.005) and lower (27.6+/-21.3%, p<0.005) in the patients than in the negative controls, respectively. The serum predominant FLC:serum creatinine ratio was elevated in parallel with an increase in CD38++ CD19- CD56+ and MPC-1+ CD45- CD49e- cells and a decrease in mature subtypes (MPC-1+ CD45+ CD49e- and MPC-1+ CD45+ CD49e+ cells), There was a significantly positive correlation between the serum predominant FLC:serum creatinine ratio and either CD38++ CD19- CD56+ (r=0.510, p<0.05) or MPC-1+ CD45- CD49e- cells (r=0.481, p<0.05). In primary AL amyloidosis M-protein is probably produced by increased monoclonal plasma cells in the bone marrow, particularly by the intermediate subpopulation with a phenotype of MPC-1+ CD45- CD49e-. PMID:16076609

Shimojima, Yasuhiro; Matsuda, Masayuki; Gono, Takahisa; Ishii, Wataru; Fushimi, Tomohisa; Hoshii, Yoshinobu; Yamada, Toshiyuki; Ikeda, Shu-Ichi

2005-03-01

352

Cancer Cell Expression of Autotaxin Controls Bone Metastasis Formation in Mouse through Lysophosphatidic Acid-Dependent Activation of Osteoclasts  

Microsoft Academic Search

BackgroundBone metastases are highly frequent complications of breast cancers. Current bone metastasis treatments using powerful anti-resorbtive agents are only palliative indicating that factors independent of bone resorption control bone metastasis progression. Autotaxin (ATX\\/NPP2) is a secreted protein with both oncogenic and pro-metastatic properties. Through its lysosphospholipase D (lysoPLD) activity, ATX controls the level of lysophosphatidic acid (LPA) in the blood.

Marion David; Estelle Wannecq; Françoise Descotes; Silvia Jansen; Blandine Deux; Johnny Ribeiro; Claire-Marie Serre; Sandra Grès; Nathalie Bendriss-Vermare; Mathieu Bollen; Simone Saez; Junken Aoki; Jean-Sébastien Saulnier-Blache; Philippe Clézardin; Olivier Peyruchaud; Erik Danen

2010-01-01

353

Development of Ewing's Sarcoma from Primary Bone Marrow Derived Mesenchymal Progenitor Cells  

Microsoft Academic Search

Ewing's sarcoma is a member of Ewing's family tumors (EFTs) and the second most common solid bone and soft tissue malignancy of children and young adults. It is associated in 85% of cases with the t(11;22)(q24:q12) chromosomal trans- location that generates fusion of the 5V segment of the EWS gene with the 3V segment of the ETS family gene FLI-1.

Luisa Cironi; Paolo Provero; Konstantinos Kaloulis; Carlos Garcia-Echeverria; Francesco Hoffmann; Andreas Trumpp; Ivan Stamenkovic

2005-01-01

354

Development of a primary mouse intestinal epithelial cell monolayer culture system to evaluate factors that modulate IgA transcytosis.  

PubMed

There is significant interest in the use of primary intestinal epithelial cells in monolayer culture to model intestinal biology. However, it has proven to be challenging to create functional, differentiated monolayers using current culture methods, likely due to the difficulty in expanding these cells. Here, we adapted our recently developed method for the culture of intestinal epithelial spheroids to establish primary epithelial cell monolayers from the colon of multiple genetic mouse strains. These monolayers contained differentiated epithelial cells that displayed robust transepithelial electrical resistance. We then functionally tested them by examining immunoglobulin A (IgA) transcytosis across Transwells. IgA transcytosis required induction of polymeric Ig receptor (pIgR) expression, which could be stimulated by a combination of lipopolysaccharide and inhibition of ?-secretase. In agreement with previous studies using immortalized cell lines, we found that tumor necrosis factor-?, interleukin (IL)-1?, IL-17, and heat-killed microbes also stimulated pIgR expression and IgA transcytosis. We used wild-type and knockout cells to establish that among these cytokines, IL-17 was the most potent inducer of pIgR expression/IgA transcytosis. Interferon-?, however, did not induce pIgR expression, and instead led to cell death. This new method will allow the use of primary cells for studies of intestinal physiology. PMID:24220295

Moon, C; VanDussen, K L; Miyoshi, H; Stappenbeck, T S

2014-07-01

355

Routine use of antibiotic laden bone cement for primary total knee arthroplasty: impact on infecting microbial patterns and resistance profiles.  

PubMed

Antibiotic-laden bone cement (ALBC) is used in primary arthroplasties throughout Europe. In North America, ALBC is only FDA approved for revision arthroplasty after periprosthetic joint infection (PJI). No article has evaluated whether infecting microbial profile and resistance has changed with the introduction of ALBC. We hypothesized that prophylactic use of ALBC in primary total knee arthroplasty (TKA) has not had a significant impact on infecting pathogens, and antibiotic resistance profiles. A retrospective cohort analysis was conducted of all PJI patients undergoing primary TKA and total hip arthroplasty (THA) between January 2000 and January 2009. No significant change in the patterns of infecting PJI pathogens, and no notable increase in percentage resistance was found among organisms grown from patients with PJI that had received prophylactic antibiotic-loaded cement in their primary joint arthroplasty. Early findings suggest that routine prophylactic use of ALBC has not led to changes in infecting pathogen profile, nor has led to the emergence of antimicrobial resistance at our institution. PMID:24418770

Hansen, Erik N; Adeli, Bahar; Kenyon, Robert; Parvizi, Javad

2014-06-01

356

Heterogeneity of (TH)phorbol 12,13-dibutyrate binding in primary mouse keratinocytes at different stages of maturation  

SciTech Connect

Mouse keratinocytes respond heterogeneously to phorbol esters with distinct subpopulations stimulated to proliferate or induced to differentiate. The maturation state of the epidermal cell at the time of exposure may determine its response. The binding of phorbol esters to primary mouse keratinocytes was studied under culture conditions selecting for proliferating cells or differentiating cells. (20-TH)-12-Deoxyphorbol 13-isobutyrate ((TH)-DPB) bound to both types of cells at one class of binding sites. The dissociation constant (Kd) for (TH)DPB in the proliferative cells was 69 nM and the binding at saturation (Bmax) was 1.3 pmol/mg of protein. The corresponding values in the differentiative cells were 96 nM and 1.5 pmol/mg of protein, respectively. In contrast to the results obtained with (TH)DPB, (20-TH)phorbol 12,13-dibutyrate ((TH)PDBU) bound to both cell types in a heterogeneous fashion. The site for (TH)DPB binding seemed to correspond to the higher affinity (TH)PDBU binding site. The major difference in the cells grown in the medium containing 1.2 mM CaCl2 was an increase in the Bmax of the lower affinity binding site with the other three parameters remaining similar. The state of epidermal differentiation thus appears to modulate the amount of the lower affinity binding sites for phorbol esters.

Dunn, J.A.; Jeng, A.Y.; Yuspa, S.H.; Blumberg, P.M.

1985-11-01

357

Purkinje cells and Bergmann glia are primary targets of the TR?1 thyroid hormone receptor during mouse cerebellum postnatal development.  

PubMed

Thyroid hormone is necessary for normal development of the central nervous system, as shown by the severe mental retardation syndrome affecting hypothyroid patients with low levels of active thyroid hormone. The postnatal defects observed in hypothyroid mouse cerebellum are recapitulated in mice heterozygous for a dominant-negative mutation of Thra, the gene encoding the ubiquitous TR?1 receptor. Using CRE/loxP-mediated conditional expression approach, we found that this mutation primarily alters the differentiation of Purkinje cells and Bergmann glia, two cerebellum-specific cell types. These primary defects indirectly affect cerebellum development in a global manner. Notably, the inward migration and terminal differentiation of granule cell precursors is impaired. Therefore, despite the broad distribution of its receptors, thyroid hormone targets few cell types that exert a predominant role in the network of cellular interactions that govern normal cerebellum maturation. PMID:24346699

Fauquier, Teddy; Chatonnet, Fabrice; Picou, Frédéric; Richard, Sabine; Fossat, Nicolas; Aguilera, Nadine; Lamonerie, Thomas; Flamant, Frédéric

2014-01-01

358

Activation of antioxidant response element in mouse primary cortical cultures with sesquiterpene lactones isolated from Tanacetum parthenium.  

PubMed

Tanacetum parthenium produces biologically active sesquiterpene lactones (SL). Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor known to activate a series of genes termed the antioxidant response element (ARE). Activation of Nrf2/ARE may be useful for the treatment of neurodegenerative disease. In this study we isolated 11 SL from T. parthenium with centrifugal partition chromatography and semipreparative HPLC. Compounds were screened in vitro for their ability to activate the ARE on primary mouse cortical cultures as well as for their toxicity towards the cultures. All SL containing the ?-methylene-?-lactone moiety were able to activate the ARE and cause cellular toxicity. The structure-activity relationship among the SL isolated indicates that the guaianolides were more active and when lacking the endoperoxide functionality less toxic then the germacranolides. PMID:22923197

Fischedick, Justin T; Standiford, Miranda; Johnson, Delinda A; De Vos, Ric C H; Todorovi?, Sla?ana; Banjanac, Tijana; Verpoorte, Rob; Johnson, Jeffrey A

2012-11-01

359

Global gene expression profiling confirms the molecular fidelity of primary tumor-based orthotopic xenograft mouse models of medulloblastoma.  

PubMed

We previously showed that primary tumor-based orthotopic xenograft mouse models of medulloblastoma replicated the histopathological phenotypes of patients' original tumors. Here, we performed global gene expression profiling of 11 patient-specific xenograft models to further determine whether the xenograft tumors were molecularly accurate during serial subtransplantations in mouse brains and whether they represented all the molecular subtypes of medulloblastoma that were recently described. Analysis of the transcriptomes of 9 pairs of matched passage I xenografts and patients' tumors revealed high correlation coefficients (r(2) > 0.95 in 5 models, > 0.9 in 3 models, and > 0.85 in 1 model) and only identified 69 genes in which expressions were altered (FDR = 0.0023). Subsequent pair-wise comparisons between passage I, III, and V xenografts from the 11 models further showed that no dramatic alterations were introduced (r(2) > 0.9 in 8 models and > 0.8 in 3 models). The genetic abnormalities of each model were then identified through comparison with control RNAs from 5 normal cerebella and 2 fetal brains. Hierarchical clustering using 3 previously published molecular signatures showed that our models span the whole spectrum of molecular subtypes, including SHH (n = 2), WNT (n = 2), and the most recently identified group C (n = 4) and group D (n = 3). In conclusion, we demonstrated that the 11 orthotopic medulloblastoma xenograft models were molecularly faithful to the primary tumors, and our comprehensive collection of molecularly distinct animal models should serve as a valuable resource for the development of new targeted therapies for medulloblastoma. PMID:22459127

Zhao, Xiumei; Liu, Zhigang; Yu, Litian; Zhang, Yujing; Baxter, Patricia; Voicu, Horatiu; Gurusiddappa, Sivashankarappa; Luan, Joseph; Su, Jack M; Leung, Hon-chiu Eastwood; Li, Xiao-Nan

2012-05-01

360

Global gene expression profiling confirms the molecular fidelity of primary tumor-based orthotopic xenograft mouse models of medulloblastoma  

PubMed Central

We previously showed that primary tumor-based orthotopic xenograft mouse models of medulloblastoma replicated the histopathological phenotypes of patients' original tumors. Here, we performed global gene expression profiling of 11 patient-specific xenograft models to further determine whether the xenograft tumors were molecularly accurate during serial subtransplantations in mouse brains and whether they represented all the molecular subtypes of medulloblastoma that were recently described. Analysis of the transcriptomes of 9 pairs of matched passage I xenografts and patients' tumors revealed high correlation coefficients (r2 > 0.95 in 5 models, > 0.9 in 3 models, and > 0.85 in 1 model) and only identified 69 genes in which expressions were altered (FDR = 0.0023). Subsequent pair-wise comparisons between passage I, III, and V xenografts from the 11 models further showed that no dramatic alterations were introduced (r2 > 0.9 in 8 models and > 0.8 in 3 models). The genetic abnormalities of each model were then identified through comparison with control RNAs from 5 normal cerebella and 2 fetal brains. Hierarchical clustering using 3 previously published molecular signatures showed that our models span the whole spectrum of molecular subtypes, including SHH (n = 2), WNT (n = 2), and the most recently identified group C (n = 4) and group D (n = 3). In conclusion, we demonstrated that the 11 orthotopic medulloblastoma xenograft models were molecularly faithful to the primary tumors, and our comprehensive collection of molecularly distinct animal models should serve as a valuable resource for the development of new targeted therapies for medulloblastoma.

Zhao, Xiumei; Liu, Zhigang; Yu, Litian; Zhang, Yujing; Baxter, Patricia; Voicu, Horatiu; Gurusiddappa, Sivashankarappa; Luan, Joseph; Su, Jack M.; Leung, Hon-chiu Eastwood; Li, Xiao-Nan

2012-01-01

361

A Case of Primary Bone Marrow B-Cell Non Hodgkin's Lymphoma with Severe Thrombocytopenia: Case Report and A Review of the Literature  

PubMed Central

A 78-year-old man presented with persistent gingival bleeding. He had low platelet count of 1.0 × 109/L without any lymphadenopathy. Bone marrow specimen showed diffusely distributed small-sized lymphocytes. Combined with immunophenotypic analysis, a diagnosis of primary bone marrow B-cell non-Hodgkin’s lymphoma was made. Thrombocytopenia was considered to be caused by autoimmune destruction of platelets.

Sahara, Naohi; Matsunaga, Takashi; Uekusa, Toshimasa; Irie, Seiji; Hatanaka, Kazuhito

2010-01-01

362

A thirteen year old female with primary T-cell rich B-cell lymphoma of bone masquerading as chronic recurrent multifocal osteomyelitis  

PubMed Central

Primary lymphoma of the bone (PLB) accounts for 2% of all non-Hodgkin's lymphomas, and until recently it had not been well characterized in literature. Most cases present in adulthood (average age 50), with localized painful lesions in the long bones, cranium, or axial skeleton. We describe a case of multifocal PLB in an adolescent female. In this case, the initial presentation, with migratory large joint polyarthralgias and bone pain, mimicked chronic recurrent multifocal osteomyelitis (CRMO). Had a biopsy not been performed the diagnosis would have been missed.

Haque, Saadiya A.; Shad, Aziza; Ozdemirli, Metin; Shanmugam, Victoria K.; Kallakury, Bhaskar

2009-01-01

363

Damaging effects of chronic low-dose methotrexate usage on primary bone formation in young rats and potential protective effects of folinic acid supplementary treatment.  

PubMed

Methotrexate (MTX) is a most commonly used anti-metabolite in cancer treatment and as an anti-rheumatic drug. While MTX chemotherapy at a high dose is known to cause bone growth defects in growing bones, effects of its chronic use at a low dose on growing skeleton remain less clear. Here, we examined effects on bone growth of long-term MTX chemotherapy at a low dose in young rats, and potential protective effects of supplementary treatment with antidote folinic acid (given ip at 1 mg/kg 6 h after MTX). After two cycles of 5 once-daily MTX injections (at 0.75 mg/kg, 5 days on/9 days off/5 days on), histological analysis showed that MTX at this dose caused significant reduction in heights of growth plate and primary spongiosa bone on day 22 compared to controls (P<0.05). In contrast, a similar dosing regimen but at a lower dose (0.4 mg/kg) caused only slight or no reduction in heights of both regions. However, after the induction phase at this 0.4 mg/kg dosing, continued use of MTX at a low dose (once weekly at 0.2 mg/kg) caused a reduction in primary spongiosa height and bone volume on weeks 9 and 14, which was associated with an increased osteoclast formation and their bone surface density as well as a decreased osteoblast bone surface density in the primary spongiosa. Folinic acid supplementation was shown able to prevent the MTX effects in the primary spongiosa. These results suggest that acute use of MTX can damage growth plate and primary bone at a high dose, but not at a low dose. However, long-term use of MTX at a low dose can reduce primary bone formation probably due to decreased osteoblastic function but increased osteoclastic formation and function, and supplementary treatment with folinic acid may be potentially useful in protecting bone growth during long-term low-dose MTX chemotherapy. PMID:18976724

Fan, Chiaming; Cool, Johanna C; Scherer, Michaela A; Foster, Bruce K; Shandala, Tetyana; Tapp, Heather; Xian, Cory J

2009-01-01

364

A Promising Model of Primary Human Immunization in Human-Scid Mouse  

Microsoft Academic Search

The engraftment of human peripheral blood mononuclear cells (Hu-PBMC) from adult donors in scid mice has been published by MOSIER et al. in 1988. The possibility to obtain a secondary human immune response in human-scid mice has also been reported but attempts to induce a primary human immune response still remain difficult to achieve.In this work, an antigen (Canine albumin)

Fataki Bombil; Jean Pierre Kints; Jean Marie Scheiff; Hervé Bazin; Dominique Latinne

1996-01-01

365

Differences in Renal Tubule Primary Cilia Length in a Mouse Model of Bardet-Biedl Syndrome  

Microsoft Academic Search

Background: Bardet-Biedl syndrome (BBS) is a heterogeneous genetic disorder that comprises numerous features, including renal cystic disease. Twelve BBS genes have been identified (BBS1–12). Although the exact functions of the BBS proteins are unknown, evidence suggests that they are involved in cilia assembly, maintenance and\\/or function. Renal