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PubMed1981

Purification and further characterization of isolated matrix vesicles from rat alveolar bone.

Extracellular matrix vesicles from rat alveolar bone were isolated by collagenase digestion and differential centrifugation. Further purification was performed by discontinuous sucrose density gradient centrifugation. Control tissues, kidney and liver, were processed according to the same procedures. Sucrose density gradient centrifugation of bone matrix vesicles revealed two peaks of enzymatic activity: "light" and "heavy" vesicle-enriched fractions. Electron micrographs revealed a higher degree of purification of the "light" rather than the "heavy" vesicle-enriched fraction. This coincided with the high levels of enzymatic activity detected in this fraction. Preparations obtained from kidney and liver had significantly lower levels of activity of alkaline phosphatase and ATPase as compared to the bone matrix vesicle fractions. There were also differences in the positions of enzyme activity peaks in the sucrose gradient fractions from the three tissues studied. Electron microscopic examination of kidney and liver fractions revealed structures larger than the purified bone matrix vesicles. In addition no electron-dense material was found within organelles from kidney and liver and they were studded with numerous ribosomes. Our observations indicate that the present method of isolation and purification yields fractions of matrix vesicles which are specific to bone and are significantly different from those obtained from kidney and liver.

Acid Phosphatase↗
PubMed2005

[Experimental study of tissue engineered bone loaded with osteointergrated dental implants].

OBJECTIVE: To investigate osteogenesis and integration of osteointergrated dental implants with marrow stromal osteoblast and cancellous bone matrix compound artificial bone (MCCAB) when embedded subcutaneously. METHODS: Osteointergrated dental implants (3 mm in diameter) were inserted into cancellous bone matrix (CBM) columns (5 mm in diameter). Marrow stromal osteoblast (MSO) were cultured and expanded in the column and on the surface. The osteointergrated dental implants loaded MSO-Alginate-CBM compound was formatted. This compound was then implanted subcutaneously in nude mice, and the osteointergrated dental implants loaded Alginate-CBM compounds were implanted as control. The compound was in the mice for 4 to 8 weeks and then harvested and assessed by means of gross observation, X-ray examination, histologic observation and computerized histomorphometry for evaluation of bone formation. RESULTS: The osteogenesis of the osteointergrated dental implants loaded MSO-Alginate-CBM compound was better than that of the the osteointergrated dental implants loaded Alginate-CBM compound. Both intramembranous and cartilaginous osteogenesis was seen but the former was predominant. A large amount of new bone formed around the implant and integrated well with the implant. In the control, only slight cartilage osteogenesis was seen and no integration was found. CONCLUSIONS: The results suggest that the new bone forms in the scaffolds and on the surface of the implant, and integration between the implant and artificial bone also occurs when they are implanted in the nude mice.

Animals↗
PubMed1979

Morphological and functional interrelationships of bone cells and matrix.

1. Bone is unique in its combination of cells, matrix and mineral. 2. It has a dual function in skeletal and mineral homeostasis. 3. Cells which are functionally different act as coordinated units depending on a variety of signals which may be hormonal, ionic, mechanical and, possibly, electrical. 4. The coordinated activity of the cells is brought about by their having an intimate structural interrelationship, by the probability of their being subject to the influence of factors contained within an extra-cellular fluid circulating around all the cells underlying the mesenchymal cell envelope, and by the possibility that the osteoclasts and osteoblasts do not represent an end-stage in differentiation. 5. The understanding of the factors which normally control cellular differentiation and function may lead to methods of reversing bone loss, rectifying the effects of metabolic bone disease and promoting the healing of fractures.

Adult↗
PubMed2003

Augmentation of rabbit posterolateral spondylodesis using a novel demineralized bone matrix-hyaluronan putty.

STUDY DESIGN: Posterolateral spinal fusion with allogeneic demineralized bone graft-hyaluronan putty in addition to autogenous iliac crest bone graft in a rabbit model. OBJECTIVES: To determine the potential efficacy of demineralized bone graft-hyaluronan putty as a bone graft enhancer. SUMMARY OF BACKGROUND DATA: Autograft bone is the material of choice for posterolateral lumbar intertransverse process fusion. Bone graft alternatives such as demineralized bone matrices that can be used as graft extenders, enhancers, or substitutes continue to be developed. METHODS: One hundred New Zealand white rabbits underwent bilateral posterolateral spinal fusion with autogenous iliac crest bone graft or bone graft with allogeneic rabbit demineralized bone graft-hyaluronan putty. The rabbits were killed 9 weeks later, and the lumbar spines were removed. Manual manipulation and fine detail radiography were used to assess spinal fusion, and computed tomographic images were used to quantify the volume of the fusion mass. RESULTS: In comparison with autograft bone alone, the fusion rates were greater when demineralized bone graft-hyaluronan putty was used as an adjunct to autogenous bone. Furthermore, the radiographic fusion rate was greater when demineralized bone graft-hyaluronan putty was used in a 2:1 ratio to autograft bone in comparison with a 1:1 ratio (P = 0.001). The addition of demineralized bone graft-hyaluronan putty to autograft bone was found to increase mineralized bone volume in a ratio-dependent manner (P < 0.05). CONCLUSIONS: Allogeneic demineralized bone matrix-hyaluronan putty enhances rabbit posterolateral spine fusion when used as an adjunct to autogenous bone graft. This new formulation of demineralized bone matrix may facilitate greater bone formation and successful fusion.

Animals↗
PubMed1990

Repair of calvarial nonunions by osteogenin, a bone-inductive protein.

Efforts were taken to determine the dose of bovine osteogenin (OG) that would induce more bone than that induced by 20 mg of rat particulate demineralized bone matrix (DBM), the amount allowed by the confines of an 8-mm rat craniotomy defect. Dose-response studies were performed for demineralized bone matrix alone and osteogenin, partially purified from bovine demineralized bone matrix, plus rat insoluble collagenous bone matrix (M). Demineralized bone matrix alone (2.5, 5.0, 10, 20, or 40 mg) or osteogenin (0.0625, 0.125, 0.250, 0.50, or 1.0 mg) plus 25 mg insoluble collagenous bone matrix was implanted into the pectoralis muscle for 3, 5, and 7 weeks. Both materials induced time- and dose-dependent formation of bone. The three highest dosages of osteogenin (plus insoluble collagenous bone matrix) induced more bone than 20 mg demineralized bone matrix and seemed to accelerate bone repair. However, when implanted into the 8-mm rat craniotomy defect for 4 weeks, 20 mg demineralized bone matrix and 0.5 mg osteogenin (plus insoluble collagenous bone matrix) induced comparable amounts of bone. These results suggest different mechanisms for bone formation in heterotopic and orthotopic sites.

Animals↗
PubMed2005

[Preparation and characterization of bovine bone collagen matrix].

A process of preparing bovine cortical bone in order to form materials suitable for biomedical xenograft implants was described. Fresh bone samples cut from the middiaphyseal region of bovine femora were obtained from a local slaughterhouse. The bovine bone collagen matrix (BBCM) of various shapes fabricated from bovine bone by defatting and deproteination procedure may be implanted surgically for various purposes. The bone cubes were first defatted in a mixture of defatting agent; subsequently, the samples were extracted to release noncollagenous proteins, followed by digestion using a proteolytic enzyme to remove the telopeptide portions of collagen and residual noncollagenous proteins. Finally,the samples were dried in vacuum, packed and sterilized by gamma irradiation. The bone specimens were characterized by a suite of analytical techniques involving FTIR spectroscopy, X-ray diffraction spectroscopy, differential scanning calorimetry (DSC), uniaxial tension mechanical tests and scanning electron microscopy (SEM). The result showed that BBCM occurred as a white structure with suitable porosity. It contains reasonable proprotion of mineral and organic components in the original osseous architecture of the bovine bone, which is beneficial to keeping the mechanic property and weaker immunogenicity; therefore, it can serve as a potential bone implantable material and extracellular matrix material in bone tissue engineering.

Animals↗
PubMed1985

Uptake of horseradish peroxidase by bone cells during endochondral bone development.

To investigate the mechanisms whereby bone cells absorb organic bone-matrix components during endochondral bone development, rat humeri were examined, employing horseradish peroxidase as a soluble protein tracer. Intravenously-injected peroxidase filled the osteoid layer and penetrated into the osteocyte lacunae and canaliculi, but did not enter the mineralized bone matrix. Whereas osteocytes rarely took up exogenous peroxidase, osteoblasts and osteoclasts actively endocytosed peroxidase in pinocytotic coated vesicles, tubular structures, and vacuoles. They also formed endocytotic vacuoles containing peroxidase in the Golgi area. The Golgi apparatus and dense bodies of these bone cells were, however, free of reaction products. Osteoclast ruffled borders were responsible for peroxidase absorption. In the osteoblast, osteocyte and osteoclast, endogenous peroxidatic reaction was detected only in mitochondria and not in other membrane-bounded vesicles and bodies. These results strongly suggest that both osteoblasts and osteoclasts participate in the resorption of bone-matrix organic components during bone remodelling.

Animals↗
PubMed1995

Bone morphogenetic proteins, bone marrow stromal cells, and mesenchymal stem cells. Maureen Owen revisited.

In postnatal mammals, there are persistent molecular signals and responding cells in bone to initiate osteogenesis and repair in response to trauma. The responding osteogenic precursor cells are of 2 categories: determined and inducible. The latter can be induced by demineralized bone matrix to form bone. Demineralized bone matrix consists of extracellular matrix and tightly associated bone morphogenetic proteins. The genes for bone morphogenetic proteins have been cloned, the recombinant proteins have been expressed, and currently their mechanism of action is being explored. Bone morphogenetic proteins are pleiotropic initiators of inducible osteogenic precursor cells. Bone morphogenetic proteins govern the 3 key steps in the osteogenic cascade: chemotaxis, mitosis, and differentiation. The receptors for bone morphogenetic proteins have been cloned and expressed and consist of 2 classes, Types I and II, that are membrane bound serine/threonine protein kinases. Bone morphogenetic proteins bind to extracellular matrix and their collaborative action on osteogenic cells culminates in the terminal differentiation of the osteoblast-osteocyte continuum. Bone morphogenetic proteins are currently on the threshold for clinical applications.

Animals↗
PubMed2006

The effect of demineralized bone matrix-calcium sulfate with vancomycin on calcaneal fracture healing and infection rates: a prospective study.

BACKGROUND: Displaced intra-articular calcaneal fractures may have a central cancellous bone defect area. We hypothesized that human demineralized bone matrix (DBM) calcium sulfate (CaSO(4)) might act as a reasonable alternative to autograft in calcaneal fractures. When combined with antibiotic powder, this bone graft substitute also may act as a local antibiotic delivery device. This is the first clinical study evaluating bone healing and complications associated with DBM-calcium sulfate bone graft substitute in the treatment of displaced intra-articular calcaneal fractures with a central cancellous bone defect. METHODS: Over a 29-month period, 33 displaced intra-articular calcaneal fractures with central cancellous defects were treated with open reduction and internal fixation (ORIF) and grafting with vancomycin/DBM-calcium sulfate bone graft substitute. Eleven fractures without bone defects were treated with ORIF only. Patient demographics, medical history, and CT fracture classification were recorded. Postoperatively, fractures were monitored every 2 weeks for healing and complications. RESULTS: The mean time to union was 8.2 weeks in the grafted, while the control group mean time to union was 10.4 weeks (p = 0.0117). Wound problems occurred in five (15%) of the 33 patients with grafting, all in type III fractures with severe soft-tissue swelling, and included two minor wound healing delays, and three serious wound problems. At a mean followup time of 22.4 months, no DBM-calcium sulfate grafted calcaneus demonstrated evidence of osteomyelitis. CONCLUSIONS: This is the first study examining human DBM-calcium sulfate bone graft substitute to treat displaced intra-articular calcaneal fractures. Based on these initial data, human DBM-calcium sulfate acted as an acceptable and safe autograft alternative in displaced intra-articular calcaneal fractures with moderate (5 cc to 10 cc) central cancellous bone defects.

Adolescent↗
PubMed1992

An electron microscopic study on the presence of proteoglycans in the mineralized matrix of rat and human compact lamellar bone.

The presence of proteoglycans (PGs) was studied in compact lamellar rat and human bone at the electron microscopic level. With the cationic dye cuprolinic blue (CB1), PGs could be demonstrated in the mineralized bone matrix. The amounts of PGs appeared to be equal in the different lamellae and osteons. More CBl-positive material was found in the outermost lamella of the cortex, in the perilacunar matrix around the osteocyte lacunae, and around the canaliculi. Enzyme digestion with chondroitinase ABC demonstrated that the CBl-positive rods consisted of PGs. These observations amplify biochemical studies in which PGs have been isolated from the mineralized bone matrix. The presence of CBl-positive rods in the mineralized matrix suggest that PGs do not have to be removed completely to make the matrix calcifiable.

Adult↗
PubMed1993

Healing segmental femoral defects in sheep using recombinant human bone morphogenetic protein.

A middiaphyseal, 2.5-cm osteoperiosteal segmental defect stabilized by plate fixation was created in the right femur of 17 sheep. Four treatment groups were included: Group I, no implant; Group II, inactive bone matrix; Group III, recombinant human bone morphogenetic protein (rhBMP-2) mixed with inactive bone matrix; and Group IV, autogeneic bone graft. Three animals had early failure of fixation, and the remaining 14 were evaluated at three months after implantation. Radiographs showed bony union of all defects treated with rhBMP-2 (six) and a lack of bony union in the negative-control groups treated with no implant (three) and inactive bone matrix without BMP (three). Both defects treated with autograft healed. New bone formation in the defect sites treated with rhBMP-2 first appeared one month after implantation and had a mean bending strength (expressed as a percentage of the contralateral femur) of 91% +/- 59% (mean +/- standard deviation) for defects treated with BMP-2, 77% +/- 34% for autograft, 9% +/- 8% for no implant, and 11% +/- 7% for inactive matrix without BMP. Three sheep treated with rhBMP-2 had their fixation plates removed at four months and were followed for one year. Their bone defect sites remained solidly healed one year after the initial operation.

Animals↗
PubMed1997

Detection of mineral density on the surface of mouse parietal bones: backscattered electron imaging of low accelerating voltage scanning electron microscopy.

Backscattered electron (BSE) imaging of scanning electron microscopy (SEM) was applied to a study on the mineral density of the bone surface. The neonatal and adult mouse parietal bones freed of the periosteum and covering cells were examined in a field emission scanning electron microscope equipped with a high sensitivity BSE detector at 1-30 kV accelerating voltages. The mineral density of the bone surface was observable in BSE images at 5 kV accelerating voltage while only the topographic structures of the surface were obtained under an accelerating voltage less than 5 kV. As the accelerating voltages increased from 5 kV, the bright areas were extended, probably due to the imaging of the calcified bone matrix under the uncalcified osteoid. The bone surface is usually divided into smooth and rough areas according to its irregularities. BSE images at 5 kV clearly showed that the smooth areas were further divided into dark and bright areas which apparently corresponded to the uncalcified osteoid and calcified bone matrix, respectively. Bright granules, about 1.0-3.0 microns in diameter, were sometimes observed at the border between the osteoid and calcified bone matrix; these granular calcified areas were regarded as the calcifying front forming the calcified bone matrix from the osteoid. The present study demonstrated that the distribution of the osteoid on the mouse parietal bone surface changes depending on age: the osteoid occupied a large area in the parietal bone surface in neonatal mice, but was small in adult mice. Thus, low accelerating voltage SEM using BSE provides new information on the distribution of the osteoid and the bone matrix calcification under both normal and pathological conditions.

Aging↗
PubMed1995

Glucocorticoids inhibit the attachment of osteoblasts to bone extracellular matrix proteins and decrease beta 1-integrin levels.

Prolonged glucocorticoid treatment causes osteoporosis in vivo and inhibits bone formation in vitro. We have previously shown that glucocorticoids inhibit calcification and alter osteoblast organization in a mineralizing bone organ culture system. In this study, the effect of glucocorticoids on osteoblast adhesion to bone matrix proteins and integrin expression was examined in primary rat osteoblasts and a transformed rat osteosarcoma-derived cell line ROS 17/2.8. After 24 h of treatment with corticosterone, these cells displayed a concentration-dependent decrease in adhesion to type I collagen and fibronectin. Adhesion was significantly decreased as early as 4 h after glucocorticoid administration. With 100 nM corticosterone treatment for 24 h, inhibition of the adhesion of ROS 17/2.8 cells and primary osteoblasts to fibronectin was 75 +/- 10% and 50 +/- 8%, and inhibition of adhesion to collagen was 31 +/- 10% and 65 +/- 5%, respectively. This effect was specific for osteoblasts, because glucocorticoids did not change the adhesion of fibroblasts. However, glucocorticoids did inhibit the adhesion of all cell types to rat osteonectin. To determine whether the change in osteoblast attachment to collagen and fibronectin was due to an alteration in integrin levels, the plasma membranes of these cells were labeled with [125I]lactoperoxidase, solubilized, and immunoprecipitated with an antibody to beta 1. A 24-h treatment with 100 nM corticosterone caused 80 +/- 2% and 64 +/- 9% decreases in beta 1 levels in primary osteoblasts and ROS 17/2.8 cells, respectively. These results were confirmed with immunofluorescence microscopy, which showed a glucocorticoid-induced decrease in beta 1 staining. Treatment of primary rat osteoblasts and ROS 17/2.8 cells for 72 h with corticosterone also decreased beta 1-integrin messenger RNA levels in a dose-dependent manner. We have demonstrated that the inhibition of integrin expression by glucocorticoids is involved in the decrease in osteoblast adhesion to bone extracellular matrix proteins. These data suggest that integrin modulation may influence osteoblast function and bone formation and, thus, contribute to glucocorticoid-induced osteoporosis.

Animals↗
PubMed2007

A bridging demineralized bone implant facilitates posterolateral lumbar fusion in New Zealand white rabbits.

STUDY DESIGN: Randomized controlled animal study. OBJECTIVE: Test the effectiveness of a single-strip demineralized bone matrix with hyaluronan and gelatin. SUMMARY OF BACKGROUND DATA: Demineralized bone matrix is widely used to augment spinal fusion, however, the effect of changing the physical characteristic and carrier is not known. METHODS: Demineralized bone matrix was extracted from the bones of New Zealand White rabbits, and combined with hyaluronan and gelatin to form solid strips. Forty-eight rabbits were randomized into a control and 2 experimental groups. In the control group, fusion was attempted with autograft bone. For group 2, demineralized bone matrix strips alone and for group 3, autograft and demineralized bone matrix strips were used. The fusion was assessed with manual manipulation and radiographs. The volume of the fusion mass was determined from computed tomographic images. RESULTS: By the manual palpation test, the fusion rates were 37.5%, 93.8%, and 100%, for groups 1-3, respectively (P < 0.05). By radiography, the control group fusion rate was 68.7% compared with 100% for the experimental groups (P < 0.05). The mean bone volumes of the fusion mass were 2142.2 +/- 318.5, 3132.9 +/- 632.1, and 4181.6 +/- 609.5 mm3 for groups 1-3, respectively (P < 0.05). CONCLUSIONS: The demineralized bone matrix-gel strip was able to function as both a bone-graft enhancer and a bone graft substitute in rabbit posterolateral spine fusion.

Animals↗
PubMed1996

Bisphosphonates inhibit the adhesion of breast cancer cells to bone matrices in vitro.

Bisphosphonates are used with increasing frequency in the management of skeletal complications in patients with breast cancer. In this paper, we have investigated whether bisphosphonates, besides their known beneficial effects on tumor-associated osteoclastic resorption, are capable of inhibiting breast cancer cell adhesion to bone matrix. For that we used two in vitro models for bone matrix (cortical bone slices and cryostat sections of trabecular bone from neonatal mouse tails). Four bone matrix-bound nitrogen-containing bisphosphonates (pamidronate, olpadronate, alendronate, and ibandronate) inhibited adhesion and spreading of breast cancer cells to bone dose-dependently, whereas etidronate and clodronate had little or no effect. Strikingly, the relative order of potency of the bisphosphonates in inhibiting the adhesion of cancer cells to cortical and trabecular bone corresponded to their relative antiresorptive potencies in vivo as well as their ranking in in vitro bone resorption assays with predictive value for their clinical efficacy. It appears that nitrogen-containing bisphosphonates alter selectively the adhesive properties of the extracellular bone matrix preventing the attachment of breast cancer cells to it. Besides the beneficial effects of bisphosphonates on tumor-induced osteoclastic resorption, the previously unrecognized effect presented in this paper makes these agents suitable for earlier pharmacologic intervention in patients with breast cancer at risk of developing bone metastases.

Animals↗
PubMed1999

Characterization of matrix-induced osteogenesis in rat calvarial bone defects: II. Origins of bone-forming cells.

Two experimental models that separated demineralized bone matrix (DBM) implants from the host bone were utilized to identify the origins of bone-forming cells in the repair of calvarial defects in rats. Rat DBM, Guanadine-HCl (Gdn-HCl) extracted insoluble residue of DBM, and Gdn-HCl extracted insoluble DBM to which the dialyzed Gdn-HCl extract was added back, were implanted in the two models which prevented cells of the adjacent host bone from participating in the repair. In addition, cells in the dura and in the subcutaneous tissue overlying the calvarial defect were locally labeled with (3)H-thymidine to identify the origins of those cells that were stimulated to divide and differentiate to osteoblasts. Histological studies of the temporal events that occurred during the healing process in these defect models, combined with (3)H-thymidine labeling demonstrated that the osteoblasts induced by DBM were initially derived from undifferentiated mesenchymal stem cells of the dura and later augmented by cells in the overlying connective tissue covering the defect, and not from cells in the cranial bone surrounding the circular defect. The cells of both dura and subcutaneous tissue were stimulated to proliferate and differentiate principally to osteoblasts and to a very much lesser extent to chondroblasts by DBM and by reconstituted components of DBM after Gdn-HCl extraction. Gdn-HCl-extracted insoluble DBM failed to induce bone or cartilage. These results indicate that the cytokines or other factors present in DBM are required to induce bone-forming cells derived from the dura and the overlying connective tissue for the repair of the calvarial defect.

Animals↗
PubMed2002

Thirty-six month follow-up of 25 patients treated with combination anorganic bovine-derived hydroxyapatite matrix (ABM)/cell-binding peptide (P-15) bone replacement grafts in human infrabony defects. I. Clinical findings.

BACKGROUND: Long-term evaluation of periodontal therapy is important for clinical decision making. METHODS: A synthetic cell-binding peptide (P-15) combined with anorganic bovine-derived hydroxyapatite bone matrix (ABM) was evaluated as a bone replacement graft in human periodontal osseous defects. Following initial preparation and reevaluation, flap surgery was performed. A variety of 1-, 2-, 3-wall bony defects were curetted and root surfaces subjected to mechanical debridement only. The bone defects were grafted with ABM/P-15, and the host flaps replaced or slightly coronally positioned. Weekly, then monthly deplaquing was performed until surgical reentry at 6 to 7 months. Patients were then followed on approximate 3-month recalls for 3 years. Twenty-five of the original 31 patients qualified for long-term evaluation in that their ABM/P-15 treated sites did not receive any additional therapy at the time of reentry. RESULTS: Significant clinical changes for the overall group of bony defects included improvement in mean clinical attachment level from 5.4 mm at surgery to 4.5 mm at the 6-month reentry to 3.8 mm at 3 years. There was also a decrease in mean probing depth from 5.3 mm at surgery to 3.1 mm at the 6-month reentry to 2.9 mm at 3 years. The mean gingival recession changed from +0.1 mm at surgery to 1.4 mm at the 6-month reentry to 0.9 mm at 3 years. All of these differences were at least P <0.05 from surgery to the 6-month reentry, and surgery to 3 years, but were not significant from reentry to 3 years via repeated measures analysis of variance. CONCLUSIONS: These favorable 3-year results with ABM/P-15 suggest that it may have a beneficial effect in the long-term clinical management of infrabony defects. Further long-term randomized controlled studies are needed to better assess the role of ABM/P-15 in long-term healing of periodontal osseous defects.

Adult↗
PubMed2003

Increased bone formation in mice lacking plasminogen activators.

UNLABELLED: Plasminogen activators tPA and uPA are involved in tissue remodeling, but their role in bone growth is undefined. Mice lacking tPA and uPA show increased bone formation and bone mass. The noncollagenous components of bone matrix are also increased, probably from defective degradation. This study underlines the importance of controlled bone matrix remodeling for normal endochondral ossification. INTRODUCTION: Proteolytic pathways are suggested to play a role in endochondral ossification. To elucidate the involvement of the plasminogen activators tPA and uPA in this process, we characterized the long bone phenotype in mice deficient in both tPA and uPA (tPA-/-:uPA-/-). MATERIALS AND METHODS: Bones of 2- to 7-day-old tPA-/-:uPA-/- and wild-type (WT) mice were studied using bone histomorphometry, electron microscopy analysis, and biochemical assessment of bone matrix components. Cell-mediated degradation of metabolically labeled bone matrix, osteoblast proliferation, and osteoblast differentiation, both at the gene and protein level, were studied in vitro using cells derived from both genotypes. RESULTS: Deficiency of the plasminogen activators led to elongation of the bones and to increased bone mass (25% more trabecular bone in the proximal tibial metaphysis), without altering the morphology of the growth plate. In addition, the composition of bone matrix was modified in plasminogen activator deficient mice, because an increased amount of proteoglycans (2x), osteocalcin (+45%), and fibronectin (+36%) was detected. Matrix degradation assays showed that plasminogen activators, by generating plasmin, participate in osteoblast-mediated degradation of the noncollagenous components of bone matrix. In addition, proliferation of primary osteoblasts derived from plasminogen activator-deficient mice was increased by 35%. Finally, osteoblast differentiation and formation of a mineralized bone matrix were enhanced in osteoblast cultures derived from tPA-/-:uPA-/- mice. CONCLUSIONS: The data presented indicate the importance of the plasminogen system in degradation of the noncollagenous components of bone matrix and suggest that the accumulation of these proteins in bone matrix--as occurs during plasminogen activator deficiency--may in turn stimulate osteoblast function, resulting in increased bone formation.

Animals↗