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PubMed1991

Correlation between mRNA levels for bone cell proteins and bone formation in long bones of maturing rats.

This report describes the relationship between bone formation and mRNA levels for selected bone proteins. Dynamic bone histomorphometry was used to measure bone formation in tibial periosteum of male rats from weanling (3 wk) to 52 wk old. Northern blot analysis of freshly isolated periosteal cells from the long bones was used to determine steady-state mRNA levels for the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAP), the bone matrix proteins osteocalcin (BGP), and prepro-alpha-2 (I) chain of type 1 precollagen (collagen), the osteoblast marker enzyme alkaline phosphatase (AP), and the osteoblast-derived signaling factor (growth factor) transforming growth factor-beta (TGF-beta). Radial growth at the tibial diaphysis achieved a maximum value in 8-wk-old rats and decreased progressively with age thereafter. This age-related decrease in the radial growth rate was initially due to reduced osteoblast activity; however, in older rats (greater than 17 wk old) reduced osteoblast number contributed to the decrease in bone formation. There was a strong correlation between the steady-state mRNA level for collagen and the periosteal bone formation rate. In contrast, the mRNA levels for the other bone proteins were more weakly correlated (TGF-beta and AP) or not correlated (BGP). These results suggest that the decreased bone matrix synthesis by periosteal cells in long bones of maturing rats is due to decreased expression of genes for bone matrix proteins.

Aging↗
PubMed2001

[Experimental study of bone repair induced by cryopreserved allograft periosteum and fetal bone composition in bone defect].

OBJECTIVE: To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. METHODS: Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2, 4, 8 and 12 weeks after transplantation. RESULTS: There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4, 8 and 12 weeks after transplantation (P < 0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. CONCLUSION: It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute.

Animals↗
PubMedDate not supplied

Wide resection of primary tumors of bone and replacement with massive bone grafts: an improved technique for transplanting allogeneic bone grafts.

The treatment by wide resection of 104 local aggressive and low grade malignant bone tumors is presented with results after follow-up for 5.0 years on the average. The series comprises 29 giant cell tumors, most of them malignant or premalignant, 25 cartilaginous tumors (12 verified chondrosarcomas and 13 chondromas suspect of malignancy), 14 cases of sarcoma, 6 of fibrosarcoma, 4 parosseal sarcomas, 3 osteosarcomas and one chordoma. The remaining 36 cases include: aneurysmal bone cysts, fibrous dysplasia, chondroblastomas, osteoblastomas, giant chondromyxoid fibroma, and hemangiomas. After resection of the affected area, the resulting defect was replaced with massive autologous or allogeneic bone or osteoarticular graft in altogether 68 cases (autologous bone transplant in 61 cases and allograft in 7). A metallic endoprosthesis was used in replacement in 8 cases. No replacement of the defect was needed in 28 cases. The technique of the allogeneic bone transplantations consisted of massive bone or bone cartilage grafts which had been stored at a low temperature, and firm osteosynthesis. In every case, allogeneic (stored) bone was supplemented with autologous cancellous bone around the juncture of graft and host bone. This supplement proved to be important for success of the operation. The overall results were good in 81% and fair in 11%. Five cases had to be amputated and 3 patients were lost to follow-up, a possible total of 8% poor results. Extensive resection with massive bone grafting is a worthwhile approach in the treatment of selected cases of aggressive and low grade malignant bone tumors.

Adolescent↗
PubMed1994

Development of hemopoietic bone marrow within the ectopic bone induced by bone morphogenetic protein.

Neogenesis of hemopoietic bone marrow was observed in the ectopic bone induced by implantation of bone morphogenetic protein (BMP) into the muscle of mice. A bone rudiment-like cartilage was formed first, which was then replaced by bone through an endochondral ossification process. Bone marrow formation occurred after the invasion of blood vessels into the cartilage, with accumulations of small lymphocyte-like cells and the appearance of large hemopoietic stem cells around the blood vessels. The stem cells proliferated and differentiated to form hemopoietic bone marrow. This process was similar to the embryogenesis of bone and bone marrow, but the hemopoietic activity of the BMP-induced bone was transient. The bone marrow was converted to fatty tissue within 2 weeks of its appearance. This study showed that implantation of BMP in the ectopic sites offers a useful model to investigate the neogenesis of ectopic bone marrow and the regulation of its activity.

Adipose Tissue↗
PubMed1977

The cellular basis of bone turnover and bone loss: a rebuttal of the osteocytic resorption--bone flow theory.

There is now sufficient evidence to conclude that the osteocytic resorption--bone flow theory of bone turnove is untenable. According to this theory bone is resorbed not from the surface by osteoclasts but from within by osteocytes, towards which bone flows through tissue space away from bone forming surfaces. The need to invoke resorption by osteocytes stems from the belief that too few osteoclasts are present to account for normal bone resoption, a belief which reflects unawareness of the enormous capacity of the osteoclast and the rapidity of its advance. The belief that osteocytes resorb substantial amounts of bone rests on invalid conclusions from indirect techniques, various artifacts of specimen processing and unawareness of the microscopic characteristics of woven bone. Osteocytes enlarge their lacunae by resorbing bone only as a prelude to resorption from the surface, the osteocyte and osteoclast working together as a resorbing unit. The belief that bone can flow is incompatible both with the physical properties of bone and with a substantial body of evidence relating to Haversian remodelling; the experimental data purporting to demonstrate such flow can all be explained by conventional concepts of bone turnover.

Animals↗
PubMed2006

In vivo study on the healing of bone defects treated with bone marrow stromal cells, platelet-rich plasma, and freeze-dried bone allografts, alone and in combination.

The repair of confined trabecular bone defects in rabbits treated by autologous bone marrow stromal cells (BMSC), platelet-rich plasma (PRP), freeze-dried bone allografts (FDBA) alone and in combination (BMSC + PRP; FDBA + BMSC; FDBA + PRP; FDBA + PRP + BMSC) was compared. A critical size defect was created in the distal part of the femurs of 48 adult rabbits. Histology and histomorphometry were used in the evaluation of healing at 2, 4, and 12 weeks after surgery. The healing rate (%) was calculated by measuring the residual bone defect area. Architecture of the newly formed bone was compared with that of bone at the same distal femur area of healthy rabbits. The defect healing rate was higher in PRP + BMSC, FDBA + PRP, FDBA + BMSC, and FDBA + PRP + BMSC treatments, while lower values were achieved with PRP treatment at all experimental times. The highest bone-healing rate at 2 weeks was achieved with FDBA + PRP + BMSC treatment, which resulted significantly different from PRP (p < 0.05) and BMSC (p < 0.05) treatments. At 4 weeks, the bone-healing rate increased except for PRP treatment. Finally, the bone-healing rate of FDBA + PRP, FDBA + BMSC, and FDBA + PRP + BMSC was significantly higher than that of PRP at 12 weeks (p < 0.05). At 12 weeks, significant differences still existed between PRP, BMSC, and FDBA groups and normal bone (p < 0.05). These results showed that the combination of FDBA, BMSC and PRP permitted an acceleration in bone healing and bone remodeling processes.

Animals↗
PubMed1993

The effects of 2-year treatment with the aminobisphosphonate alendronate on bone metabolism, bone histomorphometry, and bone strength in ovariectomized nonhuman primates.

This study examined the effect of 2 yr of treatment with the aminobisphosphonate alendronate (ALN) (0.05 or 0.25 mg/kg i.v. ALN every 2 wk) on estrogen deficiency bone loss and bone strength changes in ovariectomized (OVX) baboons (n = 7 per group) and the ALN mode of action at the tissue level. Biochemical markers of bone turnover increased in OVX animals and were maintained by ALN treatment at non-OVX levels (low dose) or below (high dose). 2 yr of treatment produced no cumulative effects on bone turnover markers. Histomorphometry showed a marked increase in cancellous bone remodeling in OVX animals. Activation frequency increased from 0.48 to 0.86 per yr (L5 vertebra), and the osteoid surfaces from 9 to 13.5% (P < 0.05). No changes were observed in eroded and osteoclast surfaces. ALN treatment decreased activation frequency and indices of bone formation to control levels (low dose) or below (high dose), did not change indices of mineralization, and increased bone mineral density (BMD) in the lumbar vertebrae (L2-L4) by 15% at 0.25 mg/kg (P < 0.05), relative to vehicle-treated animals. The mean strength of cancellous bone (L4) increased by 44% (low ALN dose) and 100% (high dose), compared with vehicle. The strength of individual bones correlated with the square of the L2-L4 BMD (r = 0.91, P < 0.0034). In conclusion, ALN treatment reversed the effects of ovariectomy on cancellous bone turnover and increased bone mass and bone strength in baboons.

Acid Phosphatase↗
PubMed1999

Enhancement by recombinant human bone morphogenetic protein-2 of bone formation by means of porous hydroxyapatite in mandibular bone defects.

Hydroxyapatite is osteoconductive and can maintain an original biocompatible form. It is useful, in the reconstruction of bone defects, to enhance the osteoconduction of hydroxyapatite with an osteogenic protein. The aim of this study was to evaluate the bone formation in surgically created defects of rabbit mandibles by a combination of recombinant human bone morphogenetic protein-2 (rhBMP-2), with porous hydroxyapatite and atelopeptide type I collagen used as the carrier for rhBMP-2. A 10-microg rhBMP-2-implanted group (n = 15) and a control group (n = 15), in which only atelopeptide type I collagen and porous hydroxyapatite were implanted, were histologically examined 3, 7, and 21 days after implantation. The alkaline phosphatase activity was also quantitatively analyzed. No new bone formation was observed in either the tested or the control group after 3 days. At 7 days, immature bone tissue was observed in some pores of the rhBMP-2implanted group, while in the control group, immature mesenchymal cells were observed. At 21 days, trabecular bone lined some pore walls. In the central portion, the bone marrow, including angioid tissue, was observed. New trabecular bone formation was observed on portions of the external surface of the hydroxyapatite disk. On the other hand, the control group showed infiltration of immature mesenchymal cells into some pores. Marginal bone formation was found in the pores close to the surface of the disk which opposed mandibular bone. The control group showed a slow, small increase in alkaline phosphatase activity in this study, while the experimental group showed a marked increase at 21 days. This increase was significantly higher in the tested group than in the control group at both 7 and 21 days. The findings indicate that rhBMP-2 accelerated bone formation by osteoconduction from porous hydroxyapatite. The combination of rhBMP-2, atelopeptide type I collagen, and porous hydroxyapatite is suggested to be advantageous for clinical application in reconstructing mandibular bone defects.

Alkaline Phosphatase↗
PubMed2005

[The effect of platelet-rich plasma combined with autogenous bone graft for bone regeneration in bone defects].

The objective of the present study was to evaluate the effect of PRP on bone healing both quantitatively and qualitatively using histomorphometrical methods in a rabbit model. The examined materials were autogenous bone, PRP alone, a mixture of autogenous bone and PRP, and whole blood (as a control). These materials were implanted into artificial defects prepared in rabbit tibiae. The observation period was set at 1, 2, 3, and 4 weeks. All specimens were used for histologic evaluation and 2- and 4-week specimens were used for histomorphometrical evaluations. The bone quantity increased when autogenous bone was applied but the percentage of mature bone in the autogenous bone site was smaller than in the PRP applied site. The results of this study suggested that the quantity of newly formed bone increased when autogenous bone was applied, but not when PRP only was applied. However, PRP might accelerate bone maturation by activating bone remodeling. According to this study, the bone quality could be altered by the application of PRP.

Animals↗
PubMed1994

Expression of mRNA of murine bone-related proteins in ectopic bone induced by murine bone morphogenetic protein-4.

To determine whether a system of ectopic bone formation induced by osteosarcoma-derived bone-inducing substance (bone morphogenetic protein-4) can be used as a model of developing bone at the molecular level, we studied the expression of bone-related protein mRNAs in the process of ectopic bone formation using non-radioisotopic in situ hybridization. Osteonectin mRNA was detected in fibroblast-like cells, which are similar to periosteal cells from the early to middle stages of bone development. The proportion of osteonectin mRNA-expressing cells was greater than that of osteopontin mRNA-expressing cells in hypertrophic chondrocytes and osteoblast-like cells. In contrast, osteopontin mRNA was localized in a limited population of hypertrophic chondrocytes, a single layer of osteoblast-like cells adjacent to the bone trabeculae in the middle stage of bone formation, and in a limited subset of osteocytes in the late stage. A strong osteocalcin mRNA signal was detected in osteoblast-like cells from the middle to late stages and in a limited subset of osteocytes in the late stage of bone development. Since the sequential gene expression pattern of bone-related proteins in the present system is comparable to that in embryonic osteogenesis, this system may be useful as a model for studying gene expression in osteogenesis.

Animals↗
PubMed2005

Attainment of peak bone mass and bone turnover rate in relation to estrous cycle, pregnancy and lactation in colony-bred Sprague-Dawley rats: suitability for studies on pathophysiology of bone and therapeutic measures for its management.

Alteration in biochemical markers of bone turnover and bone mineral density (BMD) of whole body and isolated femur and tibia in relation to age, estrous cycle, pregnancy and lactation and suitability of use of rat as model for studies on pathophysiology of bone and therapeutic measures for its management were investigated. Immature rats (1, 1.5 and 2 month of age; weighing, respectively, 39.3+/-1.0, 67.8+/-2.4 and 87.2+/-5.2 g) exhibited high rate of bone turnover, as evidenced by high serum osteocalcin and alkaline phosphatase and urine calcium/creatinine ratio. However, their BMD (whole body or of isolated long bones) was below measurable levels. Marked increase in body weight at 3 months (185.5+/-5.2 g) was associated with low serum osteocalcin and alkaline phosphatase and urine calcium/creatinine ratio. Biochemical markers and BMD attained at puberty at 3 months were maintained until 36 month of age. No significant change in serum calcium was observed with increasing age or on any of the biomarkers during estrous cycle, and BMD of femur and tibia isolated during proestrus and diestrus stages was almost similar. Onset of pregnancy was associated with significant increase in serum total alkaline phosphatase and osteocalcin levels, but serum calcium, urine calcium/creatinine ratio or BMD of whole body or isolated long bones were not significantly different from that at proestrus stage. No marked change, except increase in body weight (P<0.05), was also evident in these parameters between days 5 and 19 of pregnancy, irrespective of number of implantations in the uterus. A significant decrease in BMD of isolated femur (neck and mid-shaft regions) was observed on days 5 and 21 of lactation as compared to that during pregnancy or diestrus/proestrus stages of estrous cycle; the decrease being almost similar in females lactating two or six young ones. BMD of isolated tibia (global and region proximal to tibio-fibular separation point), though generally lower than that during cycle and pregnancy, was statistically non-significant. However, clear evidence of occurrence of osteoporosis during lactation, with decrease in BMD of >2.5 x S.D. in isolated femur (global, neck and mid-shaft) as well as tibia (global) was observed only when BMD data was analysed on T-/Z-score basis. Serum biochemical markers of bone turnover, too, were significantly increased in comparison to cyclic rats. Findings demonstrate marked increase in body weight and bone turnover during first 3 months of age, direct correlation between peak bone mass and onset of puberty at 3 months of age and increase in bone resorption rate during lactation. Finding of the study while might suggests possible use of rat as useful model for studies on bone turnover rate during lactation and post-weaning periods and extrapolation of the result to the human situation, but not in relation to ageing.

Age Factors↗
PubMed1996

Relationship between bone metabolism and plasma cytokine levels in patients at risk of post-transplantation bone disease after bone marrow transplantation.

Bone marrow transplant recipients may carry an increased risk of bone diseases, involving numerous factors that affect bone mineral metabolism. Interleukin-6 is a potent stimulator of bone resorption in vivo. The soluble fraction of interleukin-6 receptor is reported to trigger osteoclast formation by interleukin-6 in vitro. In a cross-sectional study we measured serum bone alkaline phosphatase concentrations and the urinary excretion of pyridinium cross-links in 21 patients after bone marrow transplantation, and investigated the relationship between these values and those for the plasma levels of interleukin-6 and soluble interleukin-6 receptor. Following bone marrow transplantation female--but not male--patients showed higher serum bone alkaline phosphatase values than age- and sex-adjusted controls (p < 0.05). Both female and male patients were characterized by increased urinary excretion values of pyridinium cross-links (p < 0.05). In contrast to a marked increase of interleukin-6 plasma levels (p < 0.001) no significant difference in the soluble interleukin-6 receptor levels was found between patients and apparently healthy persons (p = 0.838). Multiple regression analysis (taking into account different variables of the immunosuppressive regimen applied) revealed the plasma concentration of interleukin-6 as an independent predictor of the urinary excretion of pyridinium cross-links (p < 0.05). In conclusion, in patients following bone marrow transplantation, these findings indicate (a) an increase of bone formation in female--but not in male--patients possibly reflecting primary ovarian failure and (b) an enhancement of bone resorption possibly mediated by circulating interleukin-6.

Adult↗
PubMed2001

[Preparation of decalcified bone matrix, bone cement and bone morphogenetic protein composite].

OBJECTIVE: To investigate the most excellent composite proportion and the most convenient composite method of decalcified bone matrix (DBM), bone cement (BC) and bovine bone morphogenetic protein (bBMP) composite. METHODS: The canine DBM and the bBMP were prepared beforehand. First mix the bBMP and DBM together with the proportion of 1:25 with adsorption and directly, then add BC with different weight proportion and finally the complex material was examined by biomechanics, scan electron microscope, determination of bone inductive activity and implanted into the canine femoral defect caused by microwave-induced hyperthermia. RESULTS: The bBMP had strong bone inductive activity and had no affect for this function when combined with DBM with different method. There were irregular gaps in the composite and most of them with diameter less than 100 microns and the porosity of the composite was less than 20% when the proportion of DBM was 40%. So it was unfavorable for the new bone to form and the new blood vessels to reconstruct. The DBM and BC could not be combined together when the proportion of DBM was 80%. While the biomechanical intensity of the composite decreased and the qualities of the new bone forming and new blood vessels reconstructing improved successively when the proportion of DBM was from 50% to 75%. CONCLUSION: It is the most convenient that bBMP is combined with DBM directly while its bone inductive activity could not be affected. The composite material not only has high biomechanical intensity, but also is favorable for new bone to form and new blood vessels to reconstruct when the proportion of DBM is 50% and 60%. So it is favorable for bone repair in the places where need to bear a load. However it is also favorable for bone repair in the places where do not need to bear a load when the proportion of DBM is 75%.

Animals↗
PubMed2000

Effect of Paget's disease of bone on areal lumbar spine bone mineral density measured by DXA, and density of cortical and trabecular bone measured by quantitative CT.

Although bone density may be increased in bone that is affected by Paget's disease, density changes in cortical and trabecular bone and the effect on bone that is apparently unaffected by Paget's disease are relatively unexplored. We have investigated 81 vertebrae (28 affected, 53 unaffected) in 27 patients with Paget's disease, by dual X-ray absorptiometry (DXA) and by quantitative CT (QCT) bone density measurements of trabecular and cortical bone. DXA bone density was high (mean z-score = 1.62, p < 0.001) in vertebrae affected by Paget's disease, but not significantly different from normal in unaffected vertebrae (mean z-score = 0.07, ns). Mean QCT z-score in Paget's vertebrae was 2.07 (p = 0.009) for cortical bone and 1.37 (p = 0.008) for trabecular bone. DXA correlated with QCT cortical values in affected and unaffected bone (r = 0.8 and 0.56, respectively), and with QCT trabecular values (r = 0.72 and 0.48, respectively). There was no significant difference in the slopes for the correlations in affected or unaffected bone. Cortical QCT values are underestimated in Paget's disease compared with physical measurements of density, owing to the computer algorithm used. High DXA values may alert to the possibility of Paget's disease, especially if the value deviates from the expected normal sequence in lumbar vertebrae. Osteoporotic vertebrae may be overlooked if the average value of bone mineral density is taken in the lumbar spine without reviewing each vertebra.

Absorptiometry, Photon↗
PubMed1990

Relationships between tooth eruption, occlusion and alveolar bone resorption: cytological and cytochemical studies of bone resorption on rat incisor alveolar bone facing the enamel.

The rat labial incisor alveolar bone facing the enamel and bearing the occlusal force was examined by electron microscopy after being compared with the lingual alveolar bone by histological and scanning electron microscopic (SEM) observations. On the labial side, shallow resorptive lacunae were recognized all over the bone surface; these were mainly covered by osteoclasts and some mononuclear cells. The cement line was absent from the bone matrix. On the lingual side, residues of Sharpey's fibers, the bone formation surface and deep resorptive lacunae were observed by SEM. Histologically, bone remodeling areas showing both osteoclasts and active bone-forming osteoblasts on the bone surface, as well as many cement lines in bone matrix, were recognized. Furthermore, electron microscopic and cytochemical studies demonstrated that mononuclear cells located close to osteoclasts displayed osteoblastic characteristics such as alkaline phosphatase activity, a developed Golgi apparatus, and a rough endoplasmic reticulum. These findings indicate that continuous bone resorption occurs on the labial bone surface, while active bone remodeling occurs on the lingual surface. Even on the labial surface, osteoblastic cells close to osteoclasts seem to play an important role in the differentiation and or activation of osteoclasts.

Acid Phosphatase↗
PubMed2001

[Experimental study of bovine bone morphogenetic protein combined with sintered bone in the treatment of bone defects].

OBJECTIVE: To confirm the capability of ectopic bone induction of the bovine bone morphogenetic protein (bBMP), and investigate the bBMP combined with sintered bone (SB) in the treatment of bone defects. METHODS: bBMP was separated from the bovine cortical bone and implanted into the muscle pouches of the balb/c mouses. 4 mouses were killed at the 1st, 2nd, 3rd and 4th week, and their tissues were fixed in 4% formalin for histologic examination. The bBMP was combined with sintered bovine cancellons bone (SB), and segmental defects were made at raddi in 16 New Zealand rabbits. One side of the rabbits was implanted with bBMP-SB, and another side implanted with SB. The animals were killed in the 2nd, 4th, 6th and 8th week after operation for X-ray and histologic examination. RESULTS: One week after bBMP implantation, the chondrocyte was induced in muscle pouches, the woven bone was seen in 2 weeks, and the trabecular bone was formed in 4 weeks. In bone defects models, the X-ray and histologic examination showed that all parameters (chondrocyte induction, the mount of the trabecular bone, defects heal) were better in the animals treated with bBMP-SB than that with SB. CONCLUSION: 1. bBMP can induce ectopic bone formation. 2. bBMP-SB can improve the healing of the bone defects.

Animals↗
PubMed1993

Healing of cancellous bone osteotomy in rabbits--Part I: Regulation of bone volume and the regional acceleratory phenomenon in normal bone.

We performed a simple, undisplaced, incomplete osteotomy of the distal medial condyle of mature New Zealand White rabbits and permitted healing without fixation for 4 weeks. During the recovery period, the rabbits received the bone label calcein in their drinking water. From ground stained and unstained sections, we determined histomorphometric parameters in five zones: osteotomy gap, juxtaosteotomy, lateral condyle, metaphysis, and diaphysis. At 4 weeks, newly formed cancellous bone had obliterated the osteotomy gap. Osteotomy also stimulated new bone formation in cancellous bone of the medial condyle around the osteotomy. At that site, we found a fivefold increase in new bone without a change in bone volume (BV). Stimulation of bone formation was not observed in cancellous bone of the lateral condyle that had not been operated on or in cortical bone of the metaphysis or diaphysis. The maintenance of BV, despite increased new bone formation in the osteotomized medial femoral condyle, was probably due to a local acceleration of bone remodeling.

Animals↗
PubMed1997

Bone morphogenetic protein 2 (BMP-2) enhances BMP-3, BMP-4, and bone cell differentiation marker gene expression during the induction of mineralized bone matrix formation in cultures of fetal rat calvarial osteoblasts.

Normal bone formation is a prolonged process that is carefully regulated and involves sequential expression of growth regulatory factors by osteoblasts as they proliferate and ultimately differentiate. Since this orderly sequence of gene expression by osteoblasts suggests a cascade effect, and BMP-2 is capable of initiating and maintaining this effect, we examined the effects of BMP-2 on expression of other BMPs and compared these effects with the expression pattern of bone cell differentiation marker genes in primary cultures of fetal rat calvarial (FRC) osteoblasts. To examine the gene expression profile during bone cell differentiation and bone formation, we also examined the effects of rBMP-2 on bone formation in vivo and in vitro. rBMP-2 stimulated bone formation on the periosteal surface of mice when 500 ng/day rBMP-2 was injected subctaneously. When rBMP-2 was added to primary cultures of FRC osteoblasts, it accelerated mineralized nodule formation in a time and concentration-dependent manner (10-40 ng/ml). rBMP-2 (40 ng/ml) enhanced BMP-3 and -4 mRNA expression during the mineralization phase of primary cultures of FRC osteoblasts. Enhancement of BMP-3 and -4 mRNA expression by rBMP-2 was associated with increased expression of bone cell differentiation marker genes, alkaline phosphatase (ALP), type I collagen, osteocalcin (OC), osteopontin (OP), and bone sialoprotein (BSP). These results suggest that BMP-2 enhances expression of other BMP genes during bone cell differentiation. BMP-2 may act in a paracrine fashion in concert with other BMPs it induces to stimulate bone cell differentiation and bone formation during remodeling.

Amino Acid Sequence↗