An immuno-light- and electron-microscopic study of the expression of bone morphogenetic protein-2 during the process of ectopic bone formation in the rat

Immunolocalization of endogenous bone morphogenetic protein-2 (BMP-2) was investigated during the process of ectopic bone formation induced by recombinant human BMP-2 (rhBMP-2). Pellets consisting of 5 microg of rhBMP-2 and 6 mg of atelopeptide type I collagen (AC) were implanted into the calf muscles of 6-week-old rats. On days 7, 10, 14, 21 and 28 after implantation, tissue specimens were removed and examined immunohistochemically by light and electron microscopy after incubation with anti-BMP-2 monoclonal antibodies. Immunolocalization by light microscopy showed BMP-2 in chondrocytes at the pellet rim on days 7 and 10, in osteocyte-like cells in the chondroid matrix on day 14, and in osteocytes in the newly formed bone on days 21 and 28 after implantation. Ultrastructurally, on days 7 and 10 after implantation, immunolabelling for BMP-2 was aggregated in vesicle-like matrices released from mature chondrocytes in the chondroid matrix. On day 14, immunolabelling against BMP-2 had accumulated in vesicle-like matrices embedded in the calcified cartilage, in the cytoplasmic vacuoles of chondroclasts absorbing the matrix, and at the resorption surface of the calcified cartilage. On days 21 and 28, BMP-2 immunolabelling was seen in the osteoid layer and osteocyte lacunae. These results suggest that the chondrocytes and osteocytes induced by rhBMP-2 produce endogenous BMP-2. It seems that part of the endogenous BMP-2 that accumulated in the chondroid matrix was absorbed by chondroclasts and then participated in the osteoblastic differentiation of immature mesenchymal cells. This study indicates that, in addition to the implanted exogenous rhBMP-2, endogenous BMP-2 plays an important part in the maintenance of the bone-formation cascade during ectopic osteoinduction.     

Bone augmentation by recombinant human BMP-2 and collagen on adult rat parietal bone.

A composite of recombinant human bone morphogenetic protein-2 (rhBMP-2) and collagen was implanted beneath the cranial periosteum of 10-month-old rats to observe bone development and absorbent change of carrier collagen. The rhBMP-2/collagen onlay implant resulted in active bone formation and the augmented bone was connected directly with the original bone, whereas the collagen alone resulted in neither bone nor cartilage. The ossification process in the rhBMP-2/collagen occurred directly through bone formation, similar to intramembranous ossification. The carrier collagen fibers were found in the woven bone and were completely absorbed at 8 weeks in the presence of rhBMP-2, while the collagen alone implant remained encapsulated by a thin, fibrous connective tissue. Our results indicate that rhBMP-2/collagen is an effective material as a biological onlay implant, showing osteoinductive properties and being completely replaced by new bone. Carrier collagen not only plays a role in rhBMP-2 delivery, but also provides a cell anchorage for cell differentiation and remains as an artificial matrix in woven bone.     

Effect of phosphophoryn on rhBMP-2-induced bone formation

In the present study, dentin phosphophoryn (DPP) derived from fresh bovine dentin was evaluated as a co-factor for recombinant human bone morphogenetic protein-2 (rhBMP-2) in rhBMP-2-induced bone formation in rats. A 5 microg amount of Escherichia coli-derived rhBMP-2 variant was combined with DPP cross-linked to type I collagen (2.4 microg DPP/360 microg collagen), acting as carrier. Next, rhBMP-2/DPP/collagen composites were implanted by onlay-grafting beneath the cranial periosteum in 4-week-old Wistar rats. Rats were sacrificed at 2 and 3 weeks after implantation. Throughout the experimental period, rhBMP-2/DPP/collagen composite induced more bone formation than the rhBMP-2/collagen composite. Moreover, the degradation rate of rhBMP-2/DPP/collagen composite in rat was faster than that of rhBMP-2/collagen composite. Neither DPP/collagen composite nor collagen alone conducted bone formation even at 3 weeks postimplantation. These results indicate that the bone-inducing activity of rhBMP-2 is enhanced by DPP as a co-factor of rhBMP-2 in vivo.     

Characterization of the development of ectopic chondroid/bone matrix and chondrogenic/osteogenic cells during osteoinduction by rhBMP-2: a histochemical and ultrastructural study

OBJECTIVE: To investigate the characteristics of ectopic chondroid/bone matrix and chondrogenic/osteogenic cells induced by recombinant human bone morphogenetic protein-2 (rhBMP-2). MATERIALS AND METHODS: rhBMP-2 (5 microg) combined with atelocollagen was implanted into calf muscles of rats and removed on days 7, 10, 14, 21, or 28. Tissue sections were examined using: (i) hematoxylin/Alcian blue/Sirius red stain, (ii) enzyme histochemistry for alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase activity, (iii) immunohistochemistry for types I, II, and X collagen, and (iv) electron microscopy. RESULTS: On day 7, numerous fibroblast-like cells with ALP activity were present on the pellet rim. On day 10, chondroid matrix (CM) had formed, contained both type I collagen and proteoglycans, and often continued into the BMP pellet. On day 14, bone-like matrix formed around hypertrophic chondrocytes simultaneously with endochondral ossification. Coexpression of types I and II collagen within chondrocytes and osteocytes was observed throughout the time course of the experiment. CONCLUSION: These results suggest that fibroblast-like cells invading the pellet differentiate into chondrocytes and form CM under the scaffold of the carrier component. It appears that some chondrocytes change their phenotype to produce the bone-like matrix and remain within the endochondral bone. This process enables rapid osteogenesis to occur.     

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.     

Bone augmentation by onlay implant using recombinant human BMP-2 and collagen on adult rat skull without periosteum

The purpose of this study was to determine whether bone augmentation could be obtained by the composite of recombinant human bone morphogenetic protein-2 (rhBMP-2) and bioabsorbable atelocollagen when the periosteum was resected, and to compare the efficacy of the rhBMP-2/collagen implant and the collagen alone implant. The onlay implant was inserted into the space between the elevated galea aponeurotica and the skull without the periosteum of 10-month-old rats. The rhBMP-2/collagen implant resulted in osteoblasts differentiation under the galea at 1 week and active bone formation without a prior formation of cartilage. At 4 weeks, the bony trabeculae were interconnected and connected directly with the compact bone of the skull. Histomorphometric analysis at 4 weeks demonstrated that the rhBMP-2/collagen implant showed 92.5% in the volume of bone tissue, whereas the collagen alone showed 0%. The implanted collagen was gradually replaced by bone tissue in the presence of rhBMP-2. Our present results indicate that rhBMP-2 stimulates undifferentiated mesenchymal cells in the galea overlying the implant to proliferate and differentiate directly into osteoblasts on the carrier collagen fibers. The collagen matrix was stably placed on the skull and suitable as a substitute for rhBMP-2. The rhBMP-2/collagen onlay implant might be clinically applicable for bone augmentation even under the condition without the periosteum.     

Effect of hyperbaric oxygenation on bone induced by recombinant human bone morphogenetic protein-2

We examined the effect of hyperbaric oxygen (HBO) therapy on the osteoinductive activity of recombinant human bone morphogenetic protein-2 (rhBMP-2), 5mg of which was implanted into the calf muscle of rats using atelopeptide type I collagen as a carrier. Thirty Wistar rats were divided equally to be given HBO or act as controls. New bone formation was measured radiographically, biochemically, and histol ogically 3, 7, and 21 days after implantation. In both groups, new bone formation was found on day 21. However, there was significantly more new bone in the HBO group. In the HBO group, cartilage was present at the outer edge of the implanted material on day 7. On days 7 and 21, the local tissue alkaline phosphatase activity and calcium content in the HBO group were significantly greater than in the control group. These results suggest that HBO accelerated the activity and rate of osteoinduction by rhBM P-2.     

Effect of recombinant human bone morphogenetic protein-2, -4, and -7 on bone formation in rat calvarial defects

BACKGROUND: Currently, more than 20 bone morphogenetic proteins (BMPs) have been identified, and many trials have been carried out using recombinant human BMPs (rhBMPs) for bone tissue engineering. However, comparative analyses on bone formative activities of rhBMP using a preclinical model have been limited. Therefore, the aim of this study was to evaluate and compare the osteogenic potential of rhBMP-2, -4, and -7 delivered with absorbable collagen sponge (ACS) upon early (2 weeks) and complete (8 weeks) wound healing phases in a critical sized rat calvarial defect model. METHODS: Eight-millimeter critical sized calvarial defects were created in 30 male Sprague-Dawley rats. The animals were divided into three groups of 10 animals each. The defects were treated with 0.025 mg/ml rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS. The rats were sacrificed at either 2 (five rats) or 8 (five rats) weeks after surgery, and the results were evaluated histologically, histomorphometrically, and immunohistometrically. RESULTS: The surgical implantation of rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS resulted in enhanced local bone formation in the rat calvarial defect model at both 2 and 8 weeks. The amount of defect closure, new bone area, and bone density were similar in the three groups at each time point (P > 0.05). In terms of bone density and new bone area, there were statistically significant differences between results obtained at 2 weeks and those obtained at 8 weeks in all groups (P < 0.05). Two-way analysis of variance (ANOVA) revealed that there was no correlation between the time and conditions (P > 0.05), but time was found to have a strong influence on defect closure, new bone area, and bone density (P < 0.05). Irrespective of rhBMP type, positive immunoreactions of osteopontin (OPN) and osteocalcin (OCN) were evident at 2 and 8 weeks. Intense OPN and OCN staining was observed near the newly formed bone as well as in some cells within the new bone. CONCLUSIONS: Within the rhBMP types used, rhBMP concentration, and the observation interval, there appears to be no specific differences in bone regenerative potential. All rhBMPs used in this study may be considered effective factors for inducing bone formation.     

Induction of dentine in amputated pulp of dogs by recombinant human bone morphogenetic proteins-2 and -4 with collagen matrix

Recombinant human bone morphogenetic protein (BMP)-2, BMP-4 and transforming growth factor (TGF)-β₁ combined with collagen matrix as a carrier were examined for their effects on pulp regeneration and dentine formation. Seventy days after implantation of 2 μg of BMP-2, mineralized osteodentine-like tissue containing embedded osteodentinocytes was seen in the cavity. Unmineralized fibrous tissue and pulp-like loose connective tissue were also found in the same cavity. In teeth implanted with 660 ng of BMP-2 only unmineralized fibrous and pulp tissues were seen. In teeth with 220 ng of BMP-2 or collagen alone, pulp tissue was seen. It is therefore likely that the cavity fills with pulp tissue and that spindle-shaped cells elaborate extracellular matrix that mineralizes to be osteodentine in a dose-dependent manner. Similar osteodentine was seen in teeth implanted with 4 μg of BMP-4 and collagen. No distinct tubular dentine was formed, unlike an earlier experiment in which BMP-2 or -4 was implanted with enriched, inactivated dentine matrix. These findings suggest that both BMP-2 and -4 induce osteodentine formation if combined with collagen matrix; some other matrix component present in inactivated dentine matrix might be essential for further differentiation into odontoblasts. In teeth implanted with TGF-β₁, the carrier collagen remained in the cavity and little pulp tissue proliferation was seen, suggesting a possible inhibitory effect of TGF-β₁ in pulp regeneration. It is likely that the response to growth and differentiation factors is dependent on the state of differentiation of pulp cells.     

Histological assessment of bioengineered new bone in repairing osteoperiosteal mandibular defects in sheep using recombinant human bone morphogenetic protein-7

Numerous experimental studies have been published about osteoinductive bone morphogenetic proteins (BMPs). However, to our knowledge there has been no detailed histological study of a mandibular defect in a large mammal, reconstructed using BMPs. We describe here the histological features of rhBMP-7-induced bone in mandibular defects in sheep. METHODS: A 35 mm osteoperiosteal defect was created at the parasymphyseal region of the mandible in six adult sheep. The continuity of the mandible was maintained using a bony plate, and rhBMP-7 was applied on a type I collagen carrier. Bone labels were injected at selected time intervals during the follow-up period. The animals were killed after 3 months and bone samples were examined histologically, histomorphometrically, and by fluorescence microscopy. RESULTS AND CONCLUSIONS: We found a mixture of woven and lamellar bone that contained many cells with large nuclei. This had not reorganised to form cortical bone and the rhBMP-7-induced bone was more porous than the native bone. The newly-formed bone restored both endosteal and periosteal layers. rhBMP-7-induced bone was biocompatible and induced no ossification of soft tissue or abnormal growth of nearby vital structures. The mineral apposition rate was 1.98 microm/day (range 0.62-5.63 microm/day), a value close to that reported in humans. This suggests that BMPs have a limited effect in accelerating the rate of mineralisation, but promote the pre-mineralisation processes, and perhaps the formation of woven bone.     

Periodontal repair in dogs: effect of recombinant human bone morphogenetic protein-12 (rhBMP-12) on regeneration of alveolar bone and periodontal attachment

OBJECTIVES: Recombinant human bone morphogenetic protein-2 (rhBMP-2) has been shown to stimulate alveolar bone and cementum formation in periodontal defects but not a functionally oriented periodontal ligament (PDL). Subcutaneous and intramuscular implants of BMP-12 have been shown to induce tendon formation and ligament-like tissue. The objective of this study was to evaluate rhBMP-12 for periodontal regeneration, in particular PDL formation. METHODS: Six young adult Hound Labrador mongrel dogs were used. Routine supraalveolar periodontal defects were created around the mandibular premolar teeth. Three animals received rhBMP-12(0.04 mg/ml) in an absorbable collagen sponge (ACS) carrier vs. rhBMP-12(0.2 mg/mL)/ACS in contralateral defects. Three animals received rhBMP-12(1.0 mg/ml)/ACS vs. rhBMP-2(0.2 mg/ml)/ACS (total implant volume/defect approximately 1 ml). The animals were euthanized 8 weeks postsurgery and block biopsies were processed for histometric analysis. RESULTS: Bone regeneration appeared increased in sites receiving rhBMP-2/ACS compared to sites receiving rhBMP-12/ACS. Cementum regeneration was similar comparing sites implanted with rhBMP-2/ACS to sites implanted with rhBMP-12/ACS. In contrast, sites receiving rhBMP-12/ACS exhibited a functionally oriented PDL bridging the gap between newly formed bone and cementum whereas this was a rare observation in sites receiving rhBMP-2/ACS. Ankylosis appeared increased in sites receiving rhBMP-2/ACS compared to those receiving rhBMP-12/ACS. CONCLUSIONS: The outcomes of this study suggest that rhBMP-12 may have significant effects on regeneration of the PDL. Additional preclinical evaluation is needed to confirm these initial observations prior to clinical application.     

Intramuscular osteoinduction and bone marrow formation by the implantation of rhBMP-2 with atelopeptide type I collagen

The study examines the osteoinductive potential of recombinant human morphogenetic protein 2 (rhBMP-2) radiologically and histologically in rat calf muscle.Ten male Wistar rats were used. rhBMP-2 50 μg was implanted with atelopeptide type I collagen as carrier in a pouch in rat calf muscle (n=5), and atelopeptide type I collagen alone was implanted in a further five as control. Induction of osteogenesis at 4 weeks was investigated.In all rats in which rhBMP-2 had been implanted there was a radio-opaque shadow in the muscle in the soft tissue radiograph. No such shadows were noted in the control group. Histological examination showed bony trabeculum, osteoblasts, and vigorous bone marrow including fatty marrow and angioid tissue both at the margins and in the center of the excised lumps in the rhBMP-2 group. There were no such signs in the control group.rhBMP-2 may be capable not only of inducing the formation of bone, but also of inducing a ‘self-supporting bone organ’ in the muscle.     

Effect of a fibrin-fibronectin sealing system as a carrier for recombinant human bone morphogenetic protein-4 on bone formation in rat calvarial defects

BACKGROUND: Bone morphogenetic proteins (BMPs) have been shown to play an important role in bone formation during development and wound healing. Despite there being good prospects for BMP applications, an ideal carrier system for BMPs has yet to be determined. The purpose of this study was to evaluate the possibility of a fibrin-fibronectin sealing system (FFSS) as a carrier for recombinant human BMP-4 (rhBMP-4) and to evaluate the genuine osteoconductive potential of the FFSS in a rat calvarial defect model. METHODS: An 8-mm, calvarial, critical-size osteotomy defect was created in each of 30 male Sprague-Dawley rats. Three groups of 10 animals each received rhBMP-4 (0.025 mg/ml) in the FFSS, FFSS control, or sham-surgery control. The groups were evaluated using histologic and histometric parameters following 2- and 8-week healing intervals (five animals per group per healing interval). RESULTS: Surgical implantation of rhBMP-4/FFSS resulted in enhanced local bone formation at 2 and 8 weeks. New bone formation was also evident in the FFSS control; however, the amount of defect closure, new bone area, and bone density was significantly greater in the rhBMP-4/FFSS group (P < 0.05). At 8 weeks, the quantity of the new bone was greater than that observed at 2 weeks, and the specimens showed a more advanced stage of remodeling and consolidation in both groups (P < 0.05). Only very limited bone formation was observed in the sham-surgery control. CONCLUSION: The results of the present study indicated that the FFSS has osteoconductive potential and may be employed as a carrier for BMPs.     

三维打印β-TCP颌骨修复支架的生物学评价

目的:探讨三维打印β-磷酸三钙(β-Tricalcium Phosphate, β-TCP)颌骨修复支架的生物学特性及体内成骨作用。方法:采用自动注浆技术制作β-TCP支架,将前成骨细胞(MC35T3-E1)接种在支架上,扫描电镜(SEM)观察材料结构与细胞黏附,CCK-8法检测细胞增殖,ALP法检测碱性磷酸酶活性。将2种支架复合重组人骨形成蛋白-2(recombinant human bone morphogenetic protein-2, rhBMP-2)后植入大鼠体内,发泡法制作的β-TCP支架为对照组,6周后取材行组织学观察。结果:三维打印支架具有规则多孔的立体结构,适合细胞黏附,且增殖及分化能力均高于对照组(P<0.05)。组织学显示复合rhBMP-2后三维打印支架新骨生成量高于发泡法制作的β-TCP支架(P<0.05)。结论:三维打印TCP支架生物相容性良好,复合rhBMP-2后可异位成骨。     

Regeneration of defects in the articular cartilage in rabbit temporomandibular joints by bone morphogenetic protein-2

The purpose of this study was to investigate the therapeutic use of recombinant human bone morphogenetic protein-2 (rhBMP-2) in internally deranged temporomandibular joints (TMJ). Defects (2 mm in diameter) were created in the surface of the condylar head. Lyophilized rhBMP-2 with collagen as the carrier was implanted in the defects in different doses: rhBMP-2 15 microg (n = 5); rhBMP-2 3 microg (n = 5); rhBMP-2 0.6 microg (n = 5). In the two control groups, the defects were either filled with collagen alone (n = 5) or left untreated (n = 5). Three weeks postoperatively the sites of defects were examined under light microscopy. In the 15 micromg and the 3 microg groups, new cartilage had filled the defects; endochondral ossification was also found deep within the defect. In the 0.6 microg group, fibrous tissue was proliferating in most areas of the defect, although cartilage was also found in some parts. In the two control groups, there was either soft tissue repair only or no evidence of tissue repair. These findings suggest that BMP-2 could stimulate the repair of defects in the articular cartilage of the mandibular condyle head during the 3 weeks postoperatively. To observe the progress of endochondral ossification in more detail, it may be necessary to extend the experiment for a longer period of time. However, this study supports the contention that BMP-2 may be useful in the regeneration of cartilage in TMJ disease.     

The bone morphogenetic protein family: multifunctional cellular regulators in the embryo and adult

Originally identified as the active components within osteoinductive extracts derived from bone, the BMPs now are known to include a large family of proteins within the TGF-β superfamily of growth and differentiation factors. Members of the BMP family have been determined to be key signaling molecules in embryogenesis, in species ranging from Drosophila to humans. They are involved in delivering positional information, the development of hard tissues (both bones and teeth), as well as soft tissue types. When implanted into adult animals, several of the BMPs have been shown to initiate the complex cellular process resulting in the induction of bone through both the endochondral and intramembranous bone formation pathways. Preclinical studies have shown the ability of these factors to induce bone and heal large bony defects in a variety of models relevant to clinical problems in orthopedics, as well as the oral and maxillofacial dental areas. For example, implantation of recombinant human BMP-2 (rhBMP-2) has been shown to augment the alveolar ridge in several animal models, and when placed in a periodontal environment to restore not only new bone, but also the attachment tissues. Results from ongoing clinical studies support the ability of rhBMP-2 implants to induce physiologic bone.     

Experience with freeze-dried PGLA/HA/rhBMP-2 as a bone graft substitute.

We investigated bone induction by recombinant human bone morphogenetic protein-2 in rodents. The purpose of this study was to evaluate the osteoinductive potential of a resorbable bone substitute fabricated from freeze-dried poly(glycolic acid-co-lactic acid) (PGLA) mixed with hydroxyapatite particles incorporated with bone morphogenetic protein-2 in skull defects of rats (FD-PGLA/HA/rhBMP-2). The FD-PGLA/HA/rhBMP-2 composite or as a control, the FD-PGLA/HA by itself were implanted in skull defects (psi 8 mm) of rats. The samples were harvested at 2 or 4 weeks postoperatively and were studied radiographically and histologically. Four weeks after implantation, the FD-PGLA/HA/rhBMP-2 discs were completely replaced by newly-formed bone possessing bone marrow. In contrast, the defects implanted with FD-PGLA/HA were filled only with fibrous connective tissue. The results suggest that the FD-PGLA/HA/rhBMP-2 composite could be an optimum bone substitute with osteoinductive potential and could function as an alternative bone graft material for autogenous bone in humans.     

Bone morphogenetic protein-2 stimulates cell recruitment and cementogenesis during early wound healing

BACKGROUND: The unique action of bone morphogenetic proteins (BMPs) on mineralised tissue formation indicates that BMPs are good candidates for use in stimulating periodontal regeneration. Relatively little is known about the mechanisms of actions of BMPs during periodontal regeneration, although recent evidence from our laboratory suggests that the effects of BMPs may be profoundly influenced by various factors including root surface conditioning, delivery systems and masticatory forces. AIM: The aim of this study was to investigate the effect of rhBMP-2 on cell recruitment during periodontal regeneration using a pulse-chase technique where cells are labelled with a thymidine analogue (BrdU) (pulse) and the migration of their progeny is followed (chase) during early wound healing. The relationship between the rhBMP-2 influence on cell recruitment from the periodontal ligament (PDL) and its ability to stimulate cementogenesis was also evaluated. METHOD: The buccal aspect of the distal root of the first molar was denuded of its PDL, cementum and superficial dentine through a bony window created in the mandible of 64 Wistar rats under general anaesthesia. Test animals were treated with 10 microL of 500 microg/ml rhBMP-2 in a collagen membrane sponge (n=32) and control defects received 10 microl of saline in a collagen sponge (n=32). All animals received an intraperitoneal single pulse injection of 40 mg/kg BrdU label 2 days postoperatively. Groups of test and control animals (n=8) were killed 2 hours later on day 2 and at 4, 7 and 10 days postoperatively. Mandibles were processed for histological examination. RESULTS: The results show that rhBMP-2 had a profound effect on proliferation and migration of cells in the adjacent and deeper aspects of the PDL at 7 and 10 days post periodontal wounding (p<0.05). Significantly greater new cementum formation occurred in the test group at 10 days (p=0.03). CONCLUSION: This study shows that following periodontal wounding rhBMP-2 stimulates cell recruitment by increasing proliferation and migration of cells from the adjacent unwounded PDL into the wounded area, thus promoting periodontal regeneration by increasing new cementum formation.     

Effects of carrier release kinetics on bone morphogenetic protein-2-induced periodontal regeneration in vivo

BACKGROUND: Bone morphogenetic proteins (BMPs) have shown considerable promise as a therapeutic agent to enhance periodontal regeneration although the optimal characteristics of a suitable release system are not known. AIM: The aim of this study was to compare the effects of slow and fast degrading gelatin carriers on BMP-2-induced periodontal healing. METHOD: Recombinant human bone morphogenetic protein-2 (rhBMP-2) was incorporated into gelatin and subsequently differentially cross-linked to produce slow and fast release carrier systems. Release kinetics were confirmed in vitro, by measuring release of 125I-growth hormone from similar gelatin plugs. Effects of BMP were evaluated in surgically created rat periodontal fenestration defects which were processed for histology 10 days post-operatively. The rats were divided into 4 groups and the control defects were treated with either slow or fast degrading gelatin (CONs or CONf respectively), whilst test groups were treated with 1.25 microg rhBMP-2 in the slow or fast degrading gelatin (BMPs or BMPf respectively). RESULTS: BMPf greatly increased bone formation compared with the control (CONf) (1.67 +/- 0.65 versus 0.34 +/- 0.11 x 10(-4) m2), but no significant differences were observed with BMPs and CONs. In contrast, new cementum formation was significantly greater in the BMPs group compared with all other groups (p<0.05). CONCLUSION: Release kinetics of BMP may have important effects on the outcome of BMP-induced periodontal regeneration. New bone formation may be affected by rapid-release kinetics although further investigation is necessary to confirm this. In contrast, new cementum formation is promoted by slow release of BMP.