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PubMed2003

Status of Bone Allografting in Japan - Nation-Wide Survey of Bone Grafting Performed from 1995 through 1999.

We report the status of bone allografting in Japan on the basis of the information obtained through questionnaires performed by the Japanese Orthopaedic Association (JOA). JOA performed a nation-wide survey in 2000, in order to clarify the current status of musculoskeletal tissue grafting in the orthopaedic practices in Japan. Conducted period was for 5 years from 1995 to 1999. As the results of this survey, it had been clarified that 92,984 bone graftings, which included autografts, allografts and synthetic bone substitutes, were performed during conducted 5 years. While the allografts were used only in 3,212 cases (3%), autograftings were performed in 64,193 cases (69%), synthetic bone substitutes were used in 25,576 cases (28%) in this series. The proportion of the number of operations for use bone substitutes increased every year, whereas that autografting decreased. The proportion of the number of allografting remained almost unaltered. Of the 706 institutions which answered to have experiences of tissue grafting, only 193 (27%) performed allograft.Since Kitasato University Hospital Bone Bank was developed in 1971, we have applied to clinical while doing basic research for preserved bone allograft. When extensive bone graft is required, allograft is very useful. In Japan, however, allograft is not performed widely. The foundation of regional bone banks is expected to resolve this problem. Since excision of bone preparations from cadaver donors is not common, bone allografts are not supplied sufficiently at present. It is needed to develop a network connecting bone banks in Japan. The enlightenment activities to the ordinary people and medical institutions will also be required.

Journal Article↗
PubMed1999

Differential patterns of altered bone formation in different bone compartments in established osteoporosis.

AIM: To investigate the level of bone formation in the different bone compartments in cases of established osteoporosis, as previous work has concentrated on trabecular bone alone. METHODS: Bone formation rates were measured histomorphometrically, in the periosteal (P), cortical (C), subcortical (SC), and trabecular (T) compartments in iliac crest biopsies from 159 patients with established osteoporosis. The values were standardised using age and sex matched control data and patterns of differential change determined by analysis of parametric status (increased, normal, reduced). RESULTS: Mean bone formation was reduced in all four compartments. This was more marked (4.4/4.1 standard deviations below the mean in C/T, v 2.3/0.9 in P/SC) and more frequent (reduced in 81.5%/78.3% in T/C, v 43.3%/44% in P/SC) in the trabecular and cortical compartments than in the periosteal or subcortical bone. Parametric status was equal in trabecular and cortical bone in 85.4% of cases, and in periosteal and subcortical bone in 65.7%, but in all four compartments in only 35.1%, indicating differential alteration of bone formation in the two sets of compartments (T/C v P/SC). CONCLUSIONS: Altered trabecular bone formation is important in osteoporosis, but there are differential patterns of alteration in the other three compartments, emphasising the presence of different microenvironments in bone; thus the effect on the cortical compartment was similar to that on the trabecular, while the subcortical and periosteal compartments also showed linkage. The linkage between the two pairs was divergent, indicating different control of bone formation, with resultant different patterns of perturbation in osteoporosis.

Adult↗
PubMed2001

Impaired intramembranous bone formation during bone repair in the absence of tumor necrosis factor-alpha signaling.

Tumor necrosis factor-alpha (TNF-alpha) is known to mediate bone resorption; however, its role in osteogenesis has not been fully elucidated. In order to investigate the direct role of TNF-alpha signaling in the recruitment and differentiation of osteoblasts, two separate models of bone repair were used, marrow ablation and simple transverse fractures. These models were carried out in the tibiae of both wild-type and knock-out mice in which both TNF-alpha receptors (p55(-/-)/p75(-/-)) had been ablated. Marrow ablation is a unique model in which robust intramembranous bone formation is induced without an endochondral component, followed by remodeling and restoration of the original trabecular architecture of the bone marrow. In contrast, fracture repair proceeds concurrently through both endochondral and intramembranous processes of new bone tissue formation. In both models of bone repair, healing was delayed in the TNF-alpha receptor (p55(-/-)/p75(-/-)) deficient mice. In the marrow ablation model, young osteoblasts were recruited into the marrow space by day three in the wild-type mice, while the TNF-alpha (p55(-/-)/p75(-/-)) mice had only granulation tissue in the marrow cavity. Type I collagen and osteocalcin mRNA expressions were reduced approximately 30 and approximately 50%, respectively, of the control values in the TNF-alpha receptor ablated mice. In the fracture repair model there was almost a complete absence of the initial intramembranous bone formation on the periosteal surface in the TNF-alpha (p55(-/-)/p75(-/-)) mice. As healing progressed however, the callus tissues were greatly enlarged, and there was a delay in hypertrophy of the chondrocytes and the resorption of cartilage tissue. While during the initial period of fracture repair there was a marked reduction in the expression of both type I collagen and osteocalcin mRNAs in the TNF-alpha (p55(-/-)/p75(-/-)) mice, levels of these mRNAs were elevated by approximately 10-20% over the wild type at the later time points in the absence of endochondral resorption of the callus. The lack of inhibition of osteogenesis during endochondral resorption suggests that a different set of signals are involved in the recruitment of osteogenic cells during endochondral repair then during intramembranous bone formation. Co-culture of chondrocytes with a mesenchymal stem cell line was carried out to examine if chondrocytes themselves produced paracrine factors that promote osteogenic differentiation. These experiments demonstrated that chondrocytes do indeed produce factors that promoted osteogenic differentiation. In summary, the results presented here suggest that TNF-alpha plays a crucial role in promoting postnatal bone repair through the induction of osteoprogenitor cell recruitment or osteogenic cell activation in the context of intramembranous bone formation. These results further suggest that the signals that promote osteogenesis during endochondral bone formation are different from those involved in intramembranous bone formation.

Animals↗
PubMed2006

Evaluation of delayed-image bone scintigraphy to assess bone formation after distraction osteogenesis in dogs.

OBJECTIVE: To quantitatively assess distraction-induced bone formation in a crural lengthening model in dogs by use of delayed-image bone scintigraphy. ANIMALS: 12 mature Labrador Retrievers. PROCEDURE: Dogs were randomly allocated to 1 of 3 groups. A circular external skeletal fixation system was mounted on the right crus of each dog. Osteotomy of the distal portion of the tibia and fibula was performed in groups 1 and 2 and was followed by a lengthening procedure of 10 mm in the first group only. The third group served as sham-operated controls. Delayed-image bone scintigraphy with technetium-99m hydroxy methylene diphosphonate was performed 2, 4, and 6 weeks after surgery. Delayed-image-to-region-of-interest, delayed-image-to-crural, and delayed-image-to-femoral scintigraphic activity ratios were calculated. New bone formation was quantified by use of densitometric image analysis, and values for the scintigraphic ratios were compared. RESULTS: In the distraction and osteotomy groups, delayed-image-to-region-of-interest and delayed-image-to-crural ratios increased significantly. Although densitometric image analysis revealed increased bone formation after distraction, the region-of-interest ratios and crural ratios were similar in both groups. All dogs had increased delayed-image-to-femoral ratios. CONCLUSIONS AND CLINICAL RELEVANCE: Delayed-image bone scintigraphy ratios were not effective at differentiating between the amounts of distraction-induced bone and osteotomy-induced bone. Metabolic bone activity in the adjacent femur was increased as a consequence of circular external skeletal fixator placement. Delayed-image bone scintigraphy was not adequately sensitive to quantitatively monitor bone formation but may be useful as an early predictor of bone healing.

Animals↗
PubMed2000

The correlation of clinical lacrimal bone density and thickness, established at the time of DCR surgery, with systemic bone mineral densitometry testing.

BACKGROUND. Due to a growing concern with regard to the relationship between osteoporosis and fractures, we wished to examine the correlation of systemic bone density with lacrimal bone characteristics (thickness and density), as measured at the time of dacryocystorhinostomy (DCR). Significant correlation would suggest that oculoplastic surgeons may screen for osteoporosis during DCR. METHODS. A prospective study of the bone mineral density in patients (n=32) undergoing DCR was conducted. During DCR, the lacrimal bone thickness and density were estimated clinically. Postoperatively, the systemic bone density was measured by dual-energy x-ray absorptiometry (DEXA) scanning. The data were analyzed using Student's t-test, Pearson correlation and Pearson chi-square methods. RESULTS. Analyzed in a bivariate arrangement, significant correlation (p<0.05) was detected between the systemic bone density (as measured at two sites, the femoral head and lumbar spine) and the lacrimal bone characteristics (thickness and density). Therefore, the lower the lacrimal bone thickness or density, the lower the systemic bone density. INTERPRETATION. With the finding of significant correlation between lacrimal bone thickness and density and systemic bone density, oculoplastic surgeons can screen for osteoporosis during DCR. If low-density thin bone is encountered during DCR, the patient's general practitioner should be alerted.

Journal Article↗
PubMedDate not supplied

Potential application of high-resolution microfocus X-ray techniques for observation of bone structure and bone-implant interface.

PURPOSE: The aim of the present study was to evaluate the potential application of 2 types of microfocus x-ray units to study the bone structure around dental implants and at the bone-implant interface. MATERIALS AND METHODS: IMZ titanium implants were placed in the maxilla and mandible of a beagle dog. After implantation periods of 1, 2, and 3 months, the bone-implant interface was evaluated with microfocus x-ray computed tomography (CT) and microfocus x-ray fluoroscopy. RESULTS: Microfocus x-ray CT images of the bone-implant specimen at 3 months after implant placement revealed a clear distinction between the implant and the bone. The implant surface was partially covered with bone, and direct contact between the implant and bone could be clearly seen. Differences in degrees of calcification were identified by the differences in relative black and white intensity. Microfocus x-ray fluoroscopy also showed clear features of the bone and titanium implant The original drill hole and new bone formation could be recognized. These findings corresponded with traditional histologic observations by light microscopy. DISCUSSION AND CONCLUSION: Microfocus x-ray techniques are non-destructive and require a very short examination time. They are considered useful to observe details of the bone structure and bone-implant interface. Microfocus x-ray fluoroscope and microfocus x-ray CT techniques can provide a clear and distinguishable image of the bone-implant interface because of their high spatial resolution.

Alveolar Process↗
PubMed2001

Effect of circular motion exercise on bone modeling and bone mass in young rats: an animal model of isometric exercise.

The aims of the study are to develop a non-invasive animal model of circular motion exercise and to evaluate the effect of this type of exercise on bone turnover in young rats. The circular motion exercise simulates isometric exercise using an orbital shaker that oscillates at a frequency of 50 Hz and is capable of speeds from 0-400 rpm. A cage is fixed on top of the shaker and the animals are placed inside. When the shaker is turned on, the oscillatory movement should encourage the animals to hold on to the cage and use various muscle forces to stabilize themselves. Rats at 8 weeks of age were trained on the shaker for 6 weeks and static and dynamic histomorphometric analyses were performed for the proximal tibial metaphysis and the tibial shaft. The exercise resulted in no significant effect on animal body weight, gastrocnemius muscle weight and femoral weight. Although the bone formation rate of cancellous and cortical periosteum was increased by the exercise, trabecular bone volume was decreased. The exercise increased periosteal and marrow perimeters and the cross-sectional diameter of cortical bone from medial to lateral without a significant increase in the cortical bone area. These results suggest that circular motion exercise under force without movement or additional weight loading will cause bone-modeling drift with an increase in bone turnover to reconstruct bone shape in adaptation to the demand in strength. Since there is no additional weight loading during circular motion exercise, the net mass of bone is not increased. The bone mass lost in trabecular bone could possibly be due to a re-distribution of mineral to the cortical bone.

Journal Article↗
PubMed2002

[Bone regeneration using bone morphogenetic protein (BMP)].

Bone morphogenetic protein (BMP) is one of the most promising osteoinductive substances and is expected to be applied clinically for bone reconstruction. BMP has restored critical-size bone defects in numerous animal experiments, but the evaluation of bone formation by BMP in primates is a prerequisite for its clinical application. We attempted to restore the mandible bone defects in primates using BMP. The implantation of BMP with the carrier completely regenerated the mandible bone defects in the young primates, and the occlusal function was restored by the dental implants inserted into the regenerated bone. Although the use of BMP alone to regenerate mandible bone defects in old monkeys produced inconclusive results, the combination grafts of BMP and bone marrow, which contained osteoprogenitor cells, were successful. Furthermore, the combination of BMP and the culture-expanded cells derived form bone marrow grafts regenerate the segmental bone defects in the mandibles of old monkeys. Thus, the implantation of BMP with the BMP-responding cells could restore large bone defects even in elderly patients.

English Abstract↗
PubMed1982

Interrelationship between metacarpal bone mass and bone mineral content in nondialyzed and dialyzed patients with chronic renal failure.

Metacarpal bone mass (D2-d2)/D2 measured on radiographs, and bone mineral content (BMC) measured by photon absorptiometry, were studied in the forearms of 30 nondialyzed and 25 hemodialyzed patients with chronic failure and in a group of normal subjects. Metacarpal bone mass was reduced in nondialyzed women with chronic renal failure and the amount of bone mineral reduced both in males and females with chronic renal failure, whereas these measurements were normal in dialyzed patients with chronic renal failure. Bone mineral content in the forearm and metacarpal bone mass decreased with age, both in women with chronic renal failure and in normal women, the decrease being more pronounced in women with chronic renal failure than in normal women. No age-related decrease was seen in males. Bone mineral content corrected for the size of bone (BMC') decreased with age both in males and in females. Highly significant correlations were found between metacarpal bone mass and BMC' in nondialyzed and dialyzed patients with chronic renal failure, whereas no significant correlation could be demonstrated in normal subjects. The study indicates that changes in metacarpal bone mass and in BMC to some degree occur simultaneously in patients with chronic renal failure; an accelerated loss of bone mass and of bone mineral with age may be seen in women with chronic renal failure compared to normal women.

Adult↗
PubMedDate not supplied

Opposite bone remodeling effects of teriparatide and alendronate in increasing bone mass.

BACKGROUND: Antiresorptive agents for the treatment of osteoporosis suppress bone remodeling and reestablish bone turnover at a lower rate to reduce bone loss. Recombinant teriparatide (human parathyroid hormone 1-34) stimulates bone formation, increases bone mass, and improves bone microarchitecture. We contrasted the effects of once-daily doses of 20 mug of teriparatide and 10 mg of alendronate sodium on bone mineral density (BMD) and markers of bone turnover. METHODS: Markers of bone turnover and areal BMD were assessed in 203 postmenopausal women with osteoporosis in an 18-month randomized parallel double-blind study; volumetric BMD was measured in a subset of women. RESULTS: Teriparatide significantly increased markers of bone turnover that peaked at 6 months (serum procollagen type I N-terminal propeptide, 218%, and urinary N-telopeptide corrected for creatinine, 58%; P<.001); alendronate significantly decreased the markers at 6 months (-67% and -72%, respectively; P<.001). At 18 months, areal and volumetric spine BMDs were significantly higher with teriparatide than with alendronate (10.3% vs 5.5% [P<.001] and 19.0% vs 3.8% [P<.01], respectively). Areal femoral neck BMD was significantly higher than baseline in the teriparatide and alendronate groups (3.9% and 3.5%, respectively). There were no significant differences in trabecular femoral neck BMD between the teriparatide and alendronate groups (4.9% and 2.2%, respectively). Cortical volumetric femoral neck BMD was significantly different between the teriparatide and alendronate groups (-1.2% and 7.7%, respectively; P = .05). CONCLUSION: Two distinct options for the management of osteoporosis lead to increases in BMD by opposite mechanisms of action on bone remodeling.

Aged↗
PubMed1997

Histomorphometric and molecular biologic comparison of bioactive glass granules and autogenous bone grafts in augmentation of bone defect healing.

The applicability of bioactive glass (BG) granules as a substitute for bone grafts was tested by comparing the histologic, histomorphometric, and molecular biologic healing patterns to those of bone autografts and ungrafted bone defects in a rat model. The cellular response in defects filled with BG granules was characterized by continuous overexpression of type III collagen. Osteogenic mesenchymal cells, prior to their differentiation to osteoblasts, organized as a dense periosteumlike layer on the surface of the BG granules. By day 14 new bone formation was more extensive in autografted defects than in BG filled defects (p = 0.039). No cartilage-specific type II collagen mRNA was detectable, confirming the uniformity of intramembranous bone formation. The difference in the initiation of new bone formation was further confirmed by the mRNA analyses of the de novo production of TGF-beta 1 and type I collagen. Autografted defects demonstrated the highest levels of TGF-beta 1 and type I collagen mRNAs during the first 2 weeks of healing, whereas BG-filled defects showed biphasic expression patterns of the same genes. Spontaneous new bone formation in ungrafted bone defects was also characterized by biphasic expression of type I collagen gene. Osteonectin mRNA declined gradually over time in autografted and BG filled defects, whereas unfilled defects showed a gradual increase of osteonectin mRNA during healing. By 8 weeks, about 70% of the BG surface showed evidence of direct new bone contact. Energy-dispersing X-ray analyses confirmed the presence of silica-rich and CaP-rich zones at the bonding interface. In conclusion, the osteoconductive surface of bioactive glass granules efficiently bonds to ongrowing new bone but the material does not reach the capacity of autogenous bone graft in promotion of osteogenesis.

Animals↗
PubMed1995

Serum kinetics, bioavailability and bone scanning of 99mTc-labelled sodium olpadronate in patients with different rates of bone turnover.

The activity of olpadronate labelled with technetium-99m(99mTc) was monitored in plasma and urine samples after single oral (925 MBq 99mTc/10 mg, coadministered with 50 mg cold drug) and intravenous (925 MBq 99mTc/5 mg) administrations to two groups of patients with different rates of bone turnover. The first group comprised high bone turnover (HBTO) patients suffering from Paget's bone disease; the second group comprised patients with normal to low bone turnover (NBTO) having the diagnosis of rheumatoid arthritis and secondary osteoporosis. Kinetic variables were correlated with anthropomorphometric variables, biological markers of bone metabolism and plasma proteins. Data were also obtained after repeatedly dosing the HBTO patients. Additionally, Paget's bone and healthy bone (PB/HB) uptake before and after low-dose oral treatment were assessed by means of scintigraphy. Results showed that most of the kinetic variables did not differ between the two groups of patients, except for a greater Vss and smaller blood area under the curve AUC in the patients with HBTO. After a repeated-dose administration period, the blood AUC activity and Whole Body Retention (WBR) of the HBTO patients tended to be similar to those of the NBTO patients. In both groups, after oral dosing, the Cmax was 20 times lower than the C0.5 after i.v. injection, and the oral bioavailability ranged from 3% to 4%. Finally, the plasma t1/2 beta ranged from 9 to 14 h. Correlation coefficients were obtained from multiple regression analysis; kinetic variables showed very low correlations with anthropomorphometric measurements. In contrast the Vss and WBR were significantly correlated with serum alkaline phosphatase levels and the Vss also with urine hydroxyproline levels. Plasma protein concentration was also correlated with excretion parameters such as CLP and plasma t1/2 beta after an oral dose. Scintigraphic studies in the HBTO group allowed bone selectivity to be seen through skeletal drug uptake. The 15 Pagetic lesions analysed in the HBTO group showed a decrease in PB/HB ratio from 3.8 in the basal study to 2.7 after olpadronate administration for 30 days at the rate of 50 mg/day. In conclusion, the kinetic profile of 99mTc-labelled olpadronate, mainly AUC and WBR, showed a dependence upon bone metabolism and seemed unrelated to body size variables. HBTO patients showed a lower blood AUC but a higher Vss. Both variables may have been reflecting the fact that the drug binds selectively with calcified tissues and, in turn, with the target compartment. Scintigraphy confirmed the labelled-compound bone selectivity as a desirable feature for a bone-scanning agent.

Administration, Oral↗
PubMed1988

Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats.

Cells from fetal or neonatal skeleton can synthesize bone-like tissue in vitro. In contrast, formation of bone-like tissue in vitro by cells derived from adult animals has rarely been reported and has not been achieved using cells from bone marrow. We have explored development of bone-like tissue in vitro by bone marrow stromal cells. Marrow stromal cells obtained from 40-43-day-old Wistar rats were grown in primary culture for 7 days and then subcultured for 20-30 days. Cells were cultured in either alpha-minimal essential medium containing 15% fetal bovine serum, antibiotics, and 50 micrograms/ml ascorbic acid, or the above medium supplemented with either 10 mM Na-beta-glycerophosphate, 10(-8) M dexamethasone, or a combination of both. Cultures were examined using phase-contrast microscopy, undemineralized and demineralized tissue histology, histochemistry (for alkaline phosphatase activity), immunohistochemistry (for collagen type, osteonectin, and bone Gla-protein), scanning and transmission electron microscopy, energy dispersive X-ray microanalysis, and X-ray diffraction. Collagenous, mineralized nodules exhibiting morphological and ultrastructural characteristics similar to bone were formed in the cultures, but only in the presence of both beta-glycerophosphate and dexamethasone. Cells associated with the nodules exhibited alkaline phosphatase activity. The matrix of the nodules was composed predominantly of type-I collagen and both osteonectin and Gla-protein were present. X-ray microanalysis showed the presence of Ca and P, and X-ray diffraction indicated the mineral to be hydroxyapatite. The nodules were also examined for bone morphogenetic protein-like activity. Paired diffusion chambers containing partly demineralized nodules and fetal muscle were implanted intraperitonealy in rats. Induction of cartilage in relation to muscle was observed histologically after 40 days in the chambers. This finding provided further support for the bone-like nature of the nodules. The observations show that bone-like tissue can be synthesized in vitro by cells cultured from young-adult bone marrow, provided that the medium contains both beta-glycerophosphate and, particularly, dexamethasone.

Aging↗
PubMed1985

The usefulness of bone-marrow scintigraphy in the detection of bone metastasis from prostatic cancer.

We used a combination of bone and bone-marrow scintigraphy to study 25 patients with prostatic cancer. Of the 18 cases whose 99mTc-methylene diphosphonate (MDP) bone scans showed hot spots in the lower lumbar region of the spine and/or the pelvic bone, 8 had normal bone-marrow scintigrams. These 8 patients were subsequently shown to have senile, degenerative changes of the spine. On the other hand, in 9 of the 10 patients whose bone-marrow scintigrams showed accumulation defects, follow-up study and characteristic X-ray findings confirmed the presence of metastases. In all 6 cases with extensive bone metastases shown by 99mTc-MDP bone scintigraphy, 99mTc-sulphur-colloid bone-marrow scintigraphy showed multiple accumulation defects. In conclusion, bone-marrow scintigraphy was found to be useful in distinguishing metastatic lesions from benign degenerative changes in the cases with suspected bone involvement, as well as in evaluating equivocal lesions in the pelvis.

Aged↗
PubMed1991

In vitro bone resorption by isolated multinucleated giant cells from giant cell tumour of bone: light and electron microscopic study.

The behaviour of multinucleated giant cells (GCs), obtained from a giant cell tumour of the tibia and cultured on glass coverslips or on devitalized bone slices, was studied using light and electron microscopy. Monitoring the GCs on bone slices by phase-contrast microscopy revealed that they had removed calcified bone matrix resulting in excavation of lacunae, with subsequent lateral extension and perforation of the bone slices. Electron microscopy demonstrated for the first time that the GCs responsible for exavating lacunae had two specific membrane modifications, ruffled border and clear zone, and showed basically similar cytoplasmic fine structures to those of osteoclasts. Fluorescence images of the GCs on glass and on bone after rhodamine-conjugated phalloidin staining revealed that most of the GCs had an intensely fluorescent peripheral band composed of a number of F-actin dots called podosomes. Some GCs showed unusual arrangements of podosomes suggesting abortive attempts at GC formation. We have demonstrated that the band structure of the GCs cultured on bone is intimately involved in bone resorption. Two stromal cell types could be recognized. The predominant type, which seemed to be the only neoplastic element because of its proliferative capability, showed quite different fine structural and cytoskeletal features from the GCs. The other type, which was much less frequent and seemed not to proliferate, had morphological similarities to the GCs, and seemed to be their precursor. Importantly GCs cultured on bone and the osteoclasts share common structures for adhesion to and resorption of bone, strongly supporting the view that the GCs of the giant cell tumour of bone are potentially active bone resorbers and can be regarded as osteoclasts.

Adult↗
PubMed2003

Generation of a chimeric mouse reconstituted with green fluorescent protein-positive bone marrow cells: a useful model for studying the behavior of bone marrow cells in regeneration in vivo.

Studies have indicated that bone marrow contains both hematopoietic stem cells and mesenchymal stem cells that can differentiate into a variety of mesenchymal tissues, such as bone, cartilage, muscle, and adipose tissue. Therefore, bone marrow cells are thought to be very useful for cell and gene therapy for various diseases. However, the multipotentiality of these cells remains unclear. To address this issue, we established a chimeric model mouse stably reconstituted with green fluorescent protein (GFP)-marked bone marrow cells. We injected bone marrow cells from GFP-transgenic C57BL/6 mice 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. Microscopic examination and fluorescence-assisted cell sorter (FACS) analysis showed that bone marrow cells, including mesenchymal cells, were almost completely reconstituted with GFP+ cells 5 weeks after transplantation. FACS analysis with lineage-specific antibodies confirmed that the GFP+ cells could differentiate into all types of blood cells. To confirm the usefulness of this mouse model, we studied the role of circulating bone marrow-derived cells in healing of damaged intestine. We performed amputation and anastomosis of the jejunum 10 cm from the pyloric region of the stomach. On the third day after operation, a large number of GFP+ cells were infiltrated in the area of anastomosis, and these cells were positive for CD45 and F4/80 antigens. In 7 days, several cells became negative for CD45 and F4/80 and positive for alpha smooth muscle actin antigen, which is specific for smooth muscle. This finding suggested that bone marrow-derived cells had differentiated into smooth muscle. Because reconstituted bone marrow cells as opposed to injected bone marrow cells, behave naturally, this model is ideal for studying the multipotentiality of bone marrow cells in vivo.

Animals↗
PubMed1997

Measurement of total and local bone morphogenetic protein concentration in bone tumours.

Bone morphogenetic protein (BMP) has been shown to be one of the significant factors in the prognosis of bone tumours. In normal development BMP induces new bone formation and later takes part in fracture healing, but its function in malignant tumours is not known. In this study the concentration of bone morphogenetic protein was measured in primary bone tumours by two methods. Local staining intensity was detected immunohistologically by the avidin-biotin-peroxidase method determining the highest dilution of anti-serum against bovine bone morphogenetic protein. The total amount of BMP in a tumour sample was measured by an enzyme-linked immunosorbent assay technique after digesting the tissue with collagenase to remove proteins from the connective tissue. Immunohistochemical staining showed that bone morphogenetic protein was present in the cytoplasm and in reactive bone formed by malignant cells. The local concentration was highest in the tissue of giant cell tumours compared to chondrosarcoma, osteosarcoma and benign bone tumours. The total amount in malignant bone tumours was 2.4 times higher compared to benign bone tumours.

Adolescent↗
PubMed2001

Potential of an ultraporous beta-tricalcium phosphate synthetic cancellous bone void filler and bone marrow aspirate composite graft.

Autogenous cancellous bone is considered to be the best bone grafting material. Autogenous bone grafts provide scaffolding for osteoconduction, growth factors for osteoinduction, and progenitor stem cells for osteogenesis. However, the procurement morbidity, limited availability, and expense associated with the use of autogenous bone grafts are significant disadvantages. Allografts and xenografts lack the osteoinduction and osteogenesis properties of autogenous bone, and they introduce the potential for both transferring disease and triggering a host immune response. Synthetic bone grafts [hydroxyapatite or tricalcium phosphate (TCP)], while good platforms for osteoconduction, lack any intrinsic properties of osteoinduction and osteogenesis. A composite graft that combines synthetic scaffold with autogenous osteoprogenitor cells from bone marrow aspirate (BMA), a low-morbidity procedure, could potentially deliver the advantages of autogenous bone grafts without the disadvantages. A new ultraporous beta-TCP construct, engineered using solution-derived nano-particle technology, may prove to be an ideal carrier for BMA in such a composite. The unique, interconnected macroporosity, mesoporosity, and microporosity of this synthetic cancellous bone void filler allows it to wick in cells and nutrients via enhanced capillarity. Preliminary canine data support this expectation, demonstrating bone formation that suggests good penetration of cells and nutrients. These results suggest that BMA cells, absorbed into such a scaffold, may remain viable, thereby potentially making such a composite a true synthetic replacement for autogenous cancellous bone.

Animals↗