Bone regeneration by recombinant human bone morphogenetic protein-2 in rat mandibular defects. An experimental model of defect filling.

BACKGROUND: Bone defects and irregularities are major problems for dental implant and periodontal therapies. METHODS: We investigated whether the application of recombinant human bone morphogenetic protein-2 (rhBMP-2) induces bone formation in through-and-through bone defects in the rat mandible. A round through-and-through bone defect (5 mm in diameter) was created in the angle of the mandible on both sides of the jaw using a steel round bur in each of 8 Long-Evans rats. In the experimental group, polylactic acid-polyglycolic acid copolymer/gelatin sponge (PGS) containing rhBMP-2 (6 microg/60 microl) was inserted in the bone defect. In the control group, the same carrier without rhBMP-2 was applied in the bone defect on the opposite side. Four weeks after application, the rats were sacrificed. Step serial sections stained with hematoxylin and eosin at intervals of 200 microm were prepared in a bucco-lingual direction. The size of the bone defects and new bone formation were evaluated histometrically. RESULTS: In all cases in the experimental group, a large quantity of newly formed bone was observed. The bone defects were completely filled with new bone in 4 of 8 rats in the experimental group. In the control group, small amounts of new bone formation were observed along the border of the original mandibular bone. Histometrical analysis revealed that the amount of new bone was significantly larger in the rhBMP-2 treated sites than in the control sites (P <0.0001; paired t-test). CONCLUSIONS: These results indicate that the rhBMP-2/PGS system induced effective bone regeneration on mandibular defects in rats. This procedure may be suitable as an experimental model for bone regeneration using various growth factors and effective for alveolar ridge augmentation followed by dental implant surgery.     

Expression of bone morphogenetic protein in the course of osteoinduction by recombinant human bone morphogenetic protein-2

To clarify the mechanism of osteoinduction by recombinant human bone morphogenetic protein-2 (rhBMP-2), we examined the time-course localization of bone morphogenetic proteins (BMPs) immunostained by an anti-BMP-2 monoclonal antibody after implantation of pellets consisting of rhBMP-2 and collagen in rat calf muscle pouch. On day 3 after implantation, BMP was detected in the entire lump, and the intensity of staining for BMP around the implant on day 7 was weaker than that on day 3. The staining for BMP decreased with time and the region of staining for BMP remained more centralized in the implant. On day 10 after implantation, BMP was observed in part of the newly induced cartilage, especially around chondrocytes. On day 14 after implantation, BMP was localized in the newly induced woven bone. On day 21, BMP staining was found in osteoblasts at the surface of the newly induced bone. Especially, the staining for BMP decreased from day 10 to day 21. These results indicate that the woven bone was replaced with mature lamellar bone from day 14 to day 21. The present findings suggest that rhBMP-2 plays an important role in osteoinduction, especially at the early stage.     

Recombinant human bone morphogenetic protein-2 (rhBMP-2) in the treatment of mandibular sequelae after tumor resection

Background

Ameloblastoma is a locally aggressive tumor most often found in posterior body and angle of the mandible. Although ameloblastoma has histological characteristics of benignity, they have a high percentage of local recurrence and possible malignant development if treated improperly.

Case report

This report presents a treatment of unusual mandibular sequelae after tumor resection using recombinant human bone morphogenetic protein-2 (rhBMP-2) associated with hydroxyapatite (HA) and calcium triphosphate (TCP).

Discussion

Seven months after surgery, the patient was asymptomatic, with stable occlusion and class I, without signs of infection or rejection, and bone repair with rigidity compatible to an immature bone structure was observed. Reconstruction of large mandibular bone defect with a combination of rhBMP-2 and HA/TCP achieving a satisfactory result with less invasive and minimum morbidity has been demonstrated.     

Effect of recombinant human bone morphogenetic protein-2 on mandibular distraction osteogenesis

The purpose of this investigation was to study the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone formation of mandibular distraction osteogenesis. Six skeletally mature sheep underwent 10 mm of bilateral mandibular distraction osteogenesis via a custom-made distractor. Three micrograms of rhBMP-2 with a collagen carrier was implanted in the osteotomy site of one side of the mandible during the osteotomy phase. The contralateral side was used as the control group, and no material was implanted into the distracted area. At 10 days after the end of distraction, all animals were killed, and the distracted calluses were harvested for radiologic and histologic analysis. New bone was generated in the distracted zone in all groups. Histologic and radiologic examination showed that the new bone formation was greater in the rhBMP-2 group than in the control group. Quantitative computed tomography evaluation, however, did not demonstrate a significantly different mean bone density of the regenerates between the 2 groups. The results indicate that application of a rhBMP-2/collagen implant during the osteotomy phase of distraction osteogenesis increased bone formation but did not have a significant effect on bone density of the regenerates.     

个体化钛支架复合珊瑚羟基磷灰石和重组人骨形成蛋白2修复兔下颌骨缺损

目的通过观察个体化钛支架复合珊瑚羟基磷灰石(CHA)和重组人骨形成蛋白2(rhBMP-2)修复兔下颌骨缺损的实验效果,探讨下颌骨个体化修复再生的途径。方法以9只新西兰大白兔为实验对象,采用磨骨术建立兔下颌骨单侧缺损模型,利用计算机辅助设计/制作、快速成型技术等设计制作兔下颌骨个体化钛修复体,再将其与CHA和rhBMP-2复合应用于兔下颌骨标准化缺损修复,通过大体标本观察、骨密度检测、生物力学测试和组织学染色对下颌骨缺损的个体化再生修复效果进行研究。结果兔下颌骨解剖形态恢复理想,生物力学测试、骨密度检测及组织学观察均显示随时间点延长成骨效果呈明显上升趋势(P<0.05),钛支架内具有大量新骨形成,新生骨组织在24周时已明显成熟。结论采用快速成型技术制作的个体化钛支架复合CHA和rhBMP-2,可望通过骨再生途径实现下颌骨缺损的个体化修复重建。     

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.     

研究下颌骨侧嵴扩增的兔动物模型的建立

目的:建立下颌骨侧嵴扩增的动物模型,初探rhBMP-2/BG(recombinanthumanbonemorphogeneticprotein-2inbioglass)下颌骨表面和缺损内扩增的可行性。方法:磨牙区唇侧骨皮质表面,预备4～6个5mm骨缺损,一侧骨皮质表面和骨缺损内放置BG/rhBMP-2,另一侧作为空白对照。术后8周大体观察、组织学检查和组织学测量。结果:观测时间内所有植入体固位良好,下颌骨表面明显扩增,材料表面骨组织覆盖;组织学观察皮质骨表面和骨缺损内新骨形成,纤维组织分割BG颗粒,大量新生骨呈编织骨样结构,与BG颗粒直接结合。部分BG颗粒已降解被新骨取代,并与骨皮质表面直接结合,残留颗粒被新骨包围,新骨形成百分比明显高于BG(P<0.05)。结论:下颌骨侧嵴扩增模型可靠、有效。     

重组人骨形成蛋白-2、胶原、珊瑚复合人工骨异位诱导成骨的实验研究

目的 采用珊瑚作为载体,胶原作为缓释系统,制血出重组人骨形成蛋白－２（ｒｈＢＭＰ－２）胶原／珊瑚复合人工骨,并评价其骨诱导活性。方法 ｒｈＢＭＰ－２、胶原和珊瑚以一定的方式复合后,植入小鼠股部肌袋内,以单纯珊瑚植入用对照,术后不同取材,通过组织学方法检测骨诱导活性。结果 复合人工骨植入１周诱导软骨形成,２周形成编织骨,４周形成含骨髓的板层骨,同时珊瑚被降解吸收。结果 胶原、珊瑚是ｒｈＢＭＰ－２较理     

Effect of recombinant human bone morphogenetic protein-2 on bone formation in alveolar ridge defects in dogs

This study was designed to evaluate the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) combined with poly D, L lactic-co-glycolic acid (PLGA)/gelatin sponge complex (PGS) on the formation of bone in critically sized marginal defects of the mandible in dogs. Three months after extraction of the pre-molar teeth, rectangular bone defects (10 x 8 x 7 mm) were made in both sides of the mandible. A PGS block soaked in rhBMP-2 (400 microgram/ml) was implanted into one defect (BMP (+) group). As control, an untreated PGS block was implanted into the contralateral defect (BMP (-) group). 2, 4, 8, and 12 weeks after implantation, the defects were examined. In the BMP (+) group, newly formed bone was found in all defects from 4 weeks onward and was marked at 12 weeks. In contrast, the BMP (-) group showed no appreciable new bone formation, even at 12 weeks. Moreover, density of newly formed bone in the BMP (+) group was similar to that of the surrounding cortical bone at 12 weeks. These findings suggest that rhBMP-2/PGS is an effective bone substitute for reconstructive surgery of the dog mandible.     

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.     

钛网成型复合自体颗粒骨移植修复下颌骨缺损动物实验的组织学研究

目的研究钛网成型复合自体颗粒骨移植重建下颌骨节段性缺损的组织学变化。方法5只Beagle犬,每只动物一侧下颌骨制备长3.5cm的节段性缺损。用钛网成型重建下颌骨外形,钛钉固定。将切除的下颌骨块和髂骨剪成直径约2mm颗粒,将皮质骨和松质骨混合(按体积比约为3:1),并紧密充填在钛网内。术后6个月取移植骨做组织学检查,包括HE染色、硬组织切片荧光观察和Van-Gieson染色。结果移植骨的表层由多层平行排列的板层骨构成,板层骨皮质骨较薄,孔隙较多。板层骨下方移植骨内部呈现为众多粗大的小梁骨相互融合成网状结构,网状结构内部可见大量排列有序较致密的骨单位。结论钛网成型自体颗粒骨移植能够形成良好的骨组织学结构,有利于种植体植入,修复下颌骨节段性缺损的咬合功能。     

Effect of recombinant human bone morphogenetic protein-4 dose on bone formation in a rat calvarial defect model

BACKGROUND: Bone morphogenetic proteins (BMPs) are being evaluated for periodontal and bone regenerative therapy. The objective of this study was to evaluate the effect of recombinant human bone morphogenetic protein-4 (rhBMP-4) dose on local bone formation in a rat calvaria defect model. METHODS: Calvarial, 8 mm diameter, critical-size osteotomy defects were created in 140 male Sprague-Dawley rats. Seven groups of 20 animals each received either 1) rhBMP-4 (2.5 microg) in an absorbable collagen sponge (ACS) carrier, 2) rhBMP-4 (5 microg)/ACS, 3) rhBMP-4 (2.5 microg) in a beta-tricalcium phosphate (beta-TCP) carrier, 4) rhBMP-4 (5 microg)/beta-TCP, 5) ACS or 6) beta-TCP carrier controls, or 7) a sham-surgery control, and were evaluated by histologic and histometric parameters following a 2- or 8-week healing interval (10 animals/group/healing interval). RESULTS: Surgical implantation of rhBMP-4/ACS and rhBMP-4/beta-TCP resulted in enhanced local bone formation at both 2 and 8 weeks. Within the dose range examined, rhBMP-4 did not exhibit an appreciable dose-dependent response. Defect closure was not significantly different between the rhBMP-4/ACS and rhBMP-4/beta-TCP groups. New bone area of the rhBMP-4/ beta-TCP group was significantly greater than that of the rhBMP-4/ ACS group; however, bone density in the rhBMP-4/ACS group was significantly greater than that in the rhBMP-4/beta-TCP group at 8 weeks (P < 0.05). CONCLUSIONS: rhBMP-4 combined with ACS or beta-TCP has a significant potential to induce bone formation in the rat calvaria defect model. Within the selected rhBMP-4 dose range and observation interval, there appeared to be no meaningful differences in bone formation.     

Functional reconstruction of the non-human primate mandible using recombinant human bone morphogenetic protein-2

The purpose of this study was to evaluate the long-term functional properties of regenerated bone induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) in segmental bone defects of primate mandibles. The 30-mm defects were created in the mandibles of six young monkeys and the mandibles were fixed with titanium plates. Then 9 mg of rhBMP-2 permeating a poly-D, L-lactic-co-glycolic acid-coated gelatin sponge (PGS) was implanted into the bone defect. Dental implants were placed into the regenerated mandible 20 weeks after surgery, then suprastructures were placed and masticatory force loading was begun 8 weeks after the insertion of the dental implants. Bone formation and the quality of new bone were evaluated radiologically and histologically at 15 and 30 weeks after surgery, and 4 and 24 weeks after masticatory force loading. The resected mandibles were completely regenerated with the rhBMP-2-induced bone. Excellent remodelling and consolidation of new bone were observed after loading. This study demonstrated that the new bone induced by rhBMP-2 in large segmental defects was maintained and functional for at least 1 year. Bone regeneration induced by rhBMP-2 holds promise as a future therapy and may be an effective alternative to autogenous bone grafts for implant dentistry and reconstructive surgery.     

Reconstruction of the primate mandible with a combination graft of recombinant human bone morphogenetic protein-2 and bone marrow.

PURPOSE: This study evaluated whether recombinant human bone morphogenetic protein-2 (rhBMP-2) can be used to regenerate a resected part of the mandible in a primate model. MATERIALS AND METHODS: Segmental bone defects were created surgically in the mandible of Japanese monkeys. rhBMP-2 was suspended in a solution of polyglycolic co-lactic acid (PGLA) and lyophilized to make a BMP/PGLA complex. The rhBMP-2/PGLA complex and autogenous bone marrow in ratios of 3:0, 2.5:0.5, or 2:1 (vol:vol) were each implanted into the bone defects in 3 monkeys. Bone marrow or P(GLA alone were each implanted in 1 monkey as a control. The animals were killed 16 weeks after surgery, followed by radiologic and histologic evaluation. RESULTS: The implantation of bone marrow alone succeeded in reconstruction of the mandible, but the implantation of the rhBMP-2/PGLA complex showed only a small amount of bone formation. The combination graft of rhBMP-2/PGLA and bone marrow resulted in a greater degree of bone formation; especially the 2:1 combination showed the same result as only bone marrow implantation. CONCLUSION: The combination graft of rhBMP-2 and bone marrow, which requires only a small amount of bone marrow, was a reliable method for reconstruction of mandibular segmental defects in this animal model.     

The effect of varying the particle size of beta tricalcium phosphate carrier of recombinant human bone morphogenetic protein-4 on bone formation in rat calvarial defects

BACKGROUND: Beta tricalcium phosphate (beta-TCP) has been developed as one of the carriers of recombinant human bone morphogenetic protein (rhBMP). However, it is not known whether the particle size of beta-TCP is related to its resorption rate and the degree of bone formation. The purpose of this study was to evaluate the effect of using beta-TCP with different particle sizes on the ability of rhBMP-4 to enhance bone formation in the rat calvarial defect model. METHODS: Calvarial, 8-mm-diameter, critical-size defects were created in 100 male Sprague-Dawley rats. Five groups of 20 animals each received either rhBMP-4 (2.5 microg) using beta-TCP with a particle size of 50 to 150 microm, rhBMP-4 (2.5 microg) using beta-TCP with a particle size of 150 to 500 microm, a beta-TCP control with a particle size of 50 to 150 microm, a beta-TCP control with a particle size of 150 to 500 microm, or a sham-surgery control, respectively, and were evaluated by measuring their histologic and histometric parameters following a 2- and 8-week healing interval. RESULTS: There were no significant differences in the defect closure, new bone area, or augmented area between either the two rhBMP-4/beta-TCP groups or between the two beta-TCP control groups at 2 and 8 weeks. CONCLUSIONS: rhBMP-4 combined with either small- or large-particle beta-TCP had a significant effect on the induction of bone formation compared to either a small- or large-particle beta-TCP control or a sham-surgery control. Within the parameters of this study, varying the particle size of beta-TCP did not seem to have a significant effect on bone formation.     

Mandibular cystic defect: A composite approach with rhBMP-2 and rib graft

Objective  To report treatment of severe mandibular defect caused by Aneurysmal Bone Cyst (ABC) in a 6-year-old child, with off-label use of Recombinant Bone Morphogenetic Protein-2 (rhBMP-2) Study Design  Clinical Study. Method  After corrective segmental mandibulectomy, mandible was stabilized by precontoured titanium reconstruction plates spanning the entire defect. After confirming final diagnosis and a wait and watch approach, rhBMP-2 was inserted into mandibular defect along with conventional split rib graft. Result  New bone formation was identified at 3 months and was evident at radiographic examinations upto 5 months. Conclusion  Reconstruction of a large mandibular defect was facilitated by use of an osteoinductive factor (rhBMP-2) as a graft additive. Clinical Relevance  One-step salvage and reconstruction could be facilitated by use of an osteoinductive factor, as a graft additive, may be an alternative strategy for repair of large mandibular defects.     

Enhancement of bone formation by bone morphogenetic protein-2 during alveolar distraction: an experimental study in sheep

BACKGROUND: The purpose of this study was to perform alveolar ridge augmentation by distraction osteogenesis (DO) and to enhance bone regeneration through the use of recombinant human bone morphogenetic protein-2 (rhBMP-2), followed by implant placement. METHODS: Alveolar segmental osteotomy was performed in the mandible of 10 sheep followed by placement of 1.5 mm alveolar distraction devices. The study group was injected on the fifth day of distraction with a single dose of 10 microg rhBMP-2. Only distraction was performed in the control group. RESULTS: A mean alveolar augmentation of 12 mm was achieved. After 12 weeks of consolidation, the distraction devices were removed and biopsies were taken for histological and immunohistochemical characterization and morphometry of the newly formed bone. Titanium threaded cylindrical implants were then placed in the newly augmented bone. Radiological evaluation showed lifting of the transport segment and integration of the implants within both the transport segment and the regenerated bone. The histological study demonstrated that the association of DO and BMP resulted in increased trabecular bone size and volume (32.2%+/-0.95% versus 18.6%+/-0.71%; P <1 x 10(-17) after 24 days of lengthening and 63.8%+/-1.89% versus 42.5%+/-1.33%; P<1 x 10(-15) after 12 weeks of consolidation) and increased numbers of proliferating cell nuclear antigen stained cells (0.7+/-0.04 versus 0.47+/-0.04; P<1 x 10(-10)) compared with the DO only group. CONCLUSIONS: Alveolar distraction augments atrophic alveolar ridge and creates new bone that permits implant placement. rhBMP-2 enhances bone quality and may shorten the consolidation period of distraction allowing for earlier implant placement.     

自体骨髓基质干细胞-珊瑚羟基磷灰石修复下颌骨节段缺损的实验研究

目的应用骨髓基质干细胞（BMSCs）复合珊瑚羟基磷灰石（CHA）构建组织工程化骨，修复犬下颌骨节段性缺损。方法体外分离培养，成骨诱导扩增犬BMSCs，将第2代细胞复合CHA后修复5只犬自体下颌骨右侧3cm的节段缺损；6只犬植入单纯CHA作为对照，术后12、26、32周通过影像学、大体形态、组织学和生物力学的方法检测骨缺损的修复效果。结果BMSCs-CHA复合物生长良好。随时间延长，X线片和CT显示实验组连接处骨痂形成，实验对照组连接处始终愈合较差；32周大体观察实验组骨修复较好，组织学显示有板层骨形成，连接处骨性愈合，实验对照组有编织骨形成，连接处纤维愈合。实验组与正常对照组下颌骨力学强度差异无统计学意义。结论自体成骨诱导BMSCs复合CHA形成的组织工程化骨可修复犬下颌骨节段缺损。     

钛网成形自体颗粒骨修复下颌骨缺损并同期种植的实验研究

目的 观察犬钛网成形自体颗粒骨移植修复下颌骨节段性缺损并同期植入钛种植体后的骨愈合和骨结合情况.方法 5只Beagle犬,一侧下颌骨制备长40 mm的节段性缺损;钛网成形修复下颌骨缺损.将切除后的下颌骨和自体髂骨剪成直径约2mm颗粒,骨皮质、骨松质体积比3∶1混合,紧密充填在钛网内,将2枚纯钛种植体埋置于颗粒骨内,术后6个月处死动物.用下颌骨X线片、组织学切片、扫描电镜以及能谱分析观察钛网内颗粒骨愈合以及种植体骨结合的情况.结果 钛网成形自体颗粒骨移植重建后的下颌骨外形满意,功能正常、颗粒骨成骨良好、结构优良,未见明显骨吸收.同期植入的种植体能够与周围骨组织形成良好的骨结合,并有促进邻近骨组织结构优化的趋势.结论 钛网成形自体颗粒骨移植是一种修复下颌骨节段性缺损的好方法,当修复后下颌骨形态良好、骨质优良、骨量充足时可以同期植入种植体.