珊瑚转化多孔羟基磷灰石复合rhBMP-2人工骨的动物实验研究

目的：寻找新的骨代用材料。方法：采用珊瑚转化多孔羟基磷石（ＣＨＡ）作为移植物支架,用Ｉ型胶原作为ｒｈＢＭＰ－２的缓释载体及赋形剂,将三者复合制成新型人工骨,进行了大鼠标准颅骨缺损骨移植实验。结果：ＣＨＡ／胶原具有良好的生物相容性和骨引导作用。ｒｈＢＭＰ－２／ＣＨＡ／胶原具有骨引导和骨诱导双重特性,其成骨效应明显优于单纯ＣＨＡ／胶原植入。结论：该复合人工骨有可能成为临床修复骨缺损的有效材料。     

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。     

以胶原缓释rhBMP-2复合BMSCs及珊瑚构建组织工程骨异位成骨的实验研究

目的：观察以胶原缓释rhBMP-2复合BMSCs及珊瑚构建的组织工程骨异位成骨的能力。方法：构建3种复合支架材料：1)rhBMP-2／珊瑚；2)胶原rhBMP-2／珊瑚；3)BMSCs／胶原rhBMP-2／珊瑚。分别植入裸鼠皮下，8周后观察成骨情况，并作比较。结果：第3组材料异位成骨的能力最强，第2组次之，第1组较弱。结论：动物实验中胶原是rhBMP-2适宜的缓释载体，BMSCs对促进材料异位成骨有重要意义。     

一种新型复合人工骨修复家兔下颌骨骨缺损的扫描电镜观察

目的探讨珊瑚转化型羟基磷灰石-胶原-重组人骨形成蛋白-2(CHA-Co-rhBMP-2)复合人工骨在修复颌骨缺损上的应用价值,为临床应用提供理论依据。方法健康家兔18只,随机分为实验组和两组对照组,每组6只。在双侧下颌骨体部制造1.0cm×1.0cm贯通性骨缺损,实验组用CHA-Co-rhBMP-2复合人工骨充填修复,对照组分别用CHA-Co复合骨和单纯CHA充填修复。每组动物各取3只分别在术后6周、12周处死,进行组织学观察及统计学分析。结果与对照组相比实验组材料具有明显的骨诱导活性,6周可见明显新骨形成,12周材料大部分降解,降解区域被大量成熟骨组织所取代,骨材料界面结合紧密,无纤维结缔组织。骨密度分析结果认为骨缺损区的骨密度随时间延长有逐渐增高趋势,统计学分析实验组与对照组之间骨密度有显著性差异。较对照组具有明显优势。实验动物均无感染和不良反应出现。结论CHA-Co-rhBMP-2复合人工骨具有良好的生物相容性、降解性及骨诱导活性,是一种理想的颌骨缺损修复材料。     

煅石膏、骨形成蛋白及羟基磷灰石三元复合人工骨修复骨缺损的实验研究

以煅石膏(PLP）作为颗粒型羟基磷灰石(HA)人工骨粘接成形剂和骨形成蛋白(BMP)的载体，制成三元复合人工骨．分别用HA—bBMP、HA—PLP和单纯HA植入狗下颌骨实验性骨缺损中，采用组织学、定量组织学、免疫组织化学、X线摄片和扫描电镜观察的方法评价该复合人工骨的生物学性能。术后1，2，4，8和16周观察发现，HA—bBMP—PLP复合人工骨具有明显的骨诱导活性．PLP可充当BMP缓慢释放系统载体．增强BMP骨诱导活性，和作为颗粒型HA的粘接成形剂．使复合人工骨具有一定的可塑性和成形性，可达到准确的植入，植入后早期可有效防止HA颗粒移动。本研究证实．HA—bBMP—PLP三元复合人工骨不但可限制植入后HA颗粒的早期移动，更重要的是可以迅速增加新骨形成量．从形态和功能上大大提高了复合人工骨修复骨缺损的质量。     

珊瑚羟基磷灰石骨块移植在重度牙槽骨缺损中的应用效果评价

目的 ：评价应用珊瑚羟基磷灰石骨块治疗重度牙槽骨缺损的安全性和有效性。方法 ：选择2019年1月—2021年1月青岛大学附属医院口腔种植科就诊且种植区牙槽骨量严重不足的患者29例,其中13例进行珊瑚羟基磷灰石骨块移植（CHA骨块组）,16例进行自体骨块移植（自体骨块组）。术前（T0）、术后即刻（T1）、术后6个月（T2）、种植体植入后即刻（T3）行CBCT检查,以种植体长轴为中心,测量距离牙槽嵴顶0 mm(L1)、3 mm(L2)、6 mm(L3)的骨宽度,比较2组骨增量、骨吸收量等。记录患者植骨术后疼痛程度,创口裂开、感染、骨块坏死等并发症发生情况。采用SPSS 26.0软件包对数据进行统计学分析。结果：所有患者均成功完成植骨手术,成功植入种植体并完成最终修复,平均随访（13.28±4.34）个月。植骨术后6个月,CHA骨块组的骨增加量高于自体骨块组,CHA骨块组的骨吸收量低于自体骨块组,但差异均无统计学意义（P>0.05）;CHA骨块组疼痛反应显著低于自体骨块组（P<0.05）。结论：水平向牙槽骨重度缺损患者应用珊瑚羟基磷灰石骨块进行骨增量,可以获得种植所需骨量,安全有...     

重组人骨形成蛋白-2/珊瑚复合人工骨的动物实验研究

本研究将重组人骨形成蛋白－２（ｒｈＢＭＰ－２）和珊瑚以一定的方式复合后,植入小鼠股部肌袋和兔颅骨标准大小缺损,以单纯珊瑚植入作对照,术后不同时间取材,通过组织学方法检测其骨诱导活性和骨修复能力．结果显示：ｒｈＢＭＰ－２／珊瑚复合人工骨植入小鼠肌袋１周,诱导软骨形成,３周,形成编织骨,６周,形成含骨髓的板层骨,同时,珊瑚被部分降解吸收;复合人工骨植入兔颅骨缺损后,以引导成骨和诱导成骨双重机制完成骨修复过程,术后１２周,植入物完全被成熟的骨组织取代,其骨修复效果明显优于单纯的珊瑚．此复合人工骨具有骨传导和骨诱导活性,骨修复能力较强,是一种较理想的新型生物性植骨材料     

rhBMP-2复合生物活性玻璃诱导成骨的实验研究

目的：探讨生物活性玻璃(bioactive glass BG)对重组人骨形成蛋白-2(recombinant human bone morphogenetic protein-2,rhBMP-2)诱导骨形成的影响。方法：6只大鼠皮下袋植入rhBMP-2／BG,0．2mg 0．5mg rhBMP-2,并设空白对照组。植入后3周行组织学检查。结果：0．5mg组rhBMP-2／BG具有成骨作用,其骨化过程类似于膜内化骨的直接骨形成。结论：rhBMP-2／BG复合物能诱导新骨形成,表明BG适合作为rhBMP-2载体。     

重组人骨形成蛋白-2/珊瑚复合人工骨的研制及其诱导成骨的实验研究

目的：制备珊瑚（ｃｏｒａｌ）与重组人骨形成蛋白－２（ｒｈＢＭＰ－２）复合人工骨并测试其骨诱导活性。方法：将ｒｈＢＭＰ－２与珊瑚复合后植入鼠肌袋内。６６只小鼠随机分为３组：第１组为复合骨（１∶２０ｗ／ｗ）；第２组为复合骨（１∶４０ｗ／ｗ）；第３组为单独珊瑚。术后１、３、６周处死动物，光镜下观察，并用图象分析法作成骨定量测定，结果：术后１周，可见复合材料表面及孔洞内有软骨形成。３周出现编织骨。６周形成含骨髓的板层骨。珊瑚被部分吸收。诱导成骨的量有时间依赖性和ｒｈＢＭＰ－２剂量依赖性。结论：ｒｈＢＭＰ－２／ｃｏｒａｌ复合人工骨具有良好的诱导成骨能力；珊瑚可能是目前能使用的ｒｈＢＭＰ－２最合适的缓释载体。     

聚乳酸、自体血、rhBMP-2复合体在颌骨缺损修复中作用的实验研究

目的:研究聚乳酸、自体血、rhBMP2复合体在颌骨缺损修复中的作用,寻找一种适合临床应用促进骨再生的新型材料。方法:家兔6只,随即分为2组,在每只兔的双侧下颌体部各形成一个方块形骨缺损,随机在一侧骨缺损处填塞rhBMP-2、自体血、聚乳酸复合体材料,另一侧骨缺损处不填任何材料作为对照,于2周、6周和10周行X线检查,于6周和10周分别处死动物,取缺损处标本行光镜和扫描电镜观察对比两组成骨状况及植入材料在组织内的反应。结果:X线检查及病理结果显示试验组骨缺损愈合快,成骨速度及成骨质量明显优于对照组自然愈合的骨缺损,电镜显示该材料组织相容性好,无炎性刺激反应,可降解。结论:聚乳酸、自体血、rhBMP2复合体是一种促骨再生的新型生物复合材料,可望成为临床实际应用的修复颌骨缺损的新型材料。     

Platelet-rich plasma and fibrin as delivery systems for recombinant human bone morphogenetic protein-2

The aim of the present study was (1) to test whether or not platelet-rich plasma (PRP) or commercially available fibrin can increase bone regeneration compared with non-treated defects and (2) to test whether or not PRP or fibrin increases bone regeneration when used as a delivery system for recombinant human bone morphogenetic protein-2 (rhBMP-2). In 16 New Zealand White rabbits, four evenly distributed 6 mm diameter defects were drilled into the calvarial bone. The following five treatment modalities were randomly allocated to all 64 defects: (0) untreated control, (1) fibrin alone, (2) PRP alone, (3) fibrin with 15 microg rhBMP-2 and (4) PRP with 15 microg rhBMP-2. For the fibrin gels and the PRP containing rhBMP-2, the 15 microg rhBMP-2 was incorporated by precipitation within the matrices before their gelation. After 4 weeks, the animals were sacrificed and the calvarial bones were removed for histological preparation. The area fraction of newly formed bone was determined in vertical sections from the middle of the defect by applying histomorphometrical analysis. A mean area fraction of newly formed bone was found within the former defect of 23.4% (+/-13.5%) in the control sites, of 28.4% (+/-17.4%) in the fibrin sites and of 34.5% (+/-17.4%) in the PRP sites. The statistical analysis revealed no significant difference in bone formation between the three groups (ANOVA). Addition of 15 microg rhBMP-2 in the fibrin gel (59.9+/-20.3%) and the PRP gels (63.1+/-25.3%) increased bone formation significantly. No significant difference was observed between sites, where PRP or fibrin has been used as a delivery system for rhBMP-2 (ANOVA). In conclusion, the application of fibrin gels or PRP gels to bone defects is not superior to leaving the defect untreated. Regarding the amount of bone formation, the application of 15 microg rhBMP-2 in bone defects enhances the healing significantly at 4 weeks. In this animal model, commercially available fibrin and autologous PRP gels are equally effective as delivery systems for rhBMP-2.     

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.     

胰岛素样生长因子-Ⅰ和骨形态发生蛋白-2对人牙周膜细胞增殖的作用

目的：重组人胰岛素样生长因子－Ｉ（ｒｈＩＧＦ－Ｉ）、重组人骨形态发生蛋白－２（ｒｈＢＭＰ－２）分别或联合应用对人牙周膜（ＰＤＬ）细胞增殖的影响。方法：采用组织块法体外培养人ＰＤＬ细胞,ＭＴＴ法测定ＰＤＬ细胞在不同生长因子刺激下的增殖情况。结果：ｒｈＩＧＦ－Ｉ、ｒｈＢＭＰ－２都可促进人ＰＤＬ细胞的增殖,这种促增殖作用呈一定的浓度依赖性,ｒｈＩＧＦ－Ｉ与ｒｈＢＭＰ－２联合应用对人ＰＤＬ细胞的增殖有协同作用,且与单独应用相比相差显著。结论：ｒｈＩＧＦ－Ｉ、ｒｈＢＭＰ－２可望作为牙周再生的生物活性介质,ｒｈＩＧＦ－Ｉ与ｒｈＢＭＰ－２联合应用对ＰＤＬ细胞的促增殖作用更强。     

Molded bone augmentation by a combination of barrier membrane and recombinant human bone morphogenetic protein-2

OBJECTIVES: To provide the histological background to a new method of local bone augmentation, we examined the events occurring beneath a barrier membrane applied with recombinant human bone morphogenetic protein-2 (rhBMP-2). MATERIALS AND METHODS: The effects on bone augmentation of rhBMP-2, applied with a membrane mold (BMP-Memb), over surgically-induced bone defects in rat calvaria were examined histologically, and the results compared with those from application of rhBMP-2 (BMP) alone, or of a molded membrane (Memb) alone. RESULTS: At postoperative week 2, the BMP group showed the most marked bone formation. However, the bone diminished in size by week 8. The Memb group showed slow but continuous bone formation by week 8. In the BMP-Memb group, bone filled the space in the mold at week 2, and this was maintained until week 8. Moreover, the soft tissue that had intervened between newly formed bone and the membrane in the Memb group was not evident in the BMP-Memb group, in which bone had formed directly on the membrane. CONCLUSIONS: The results suggest that the combination of rhBMP-2 and barrier membrane has advantages in producing and maintaining bone in the intended shape by inducing osteoblasts directly on the inner surface of the membrane.     

骨形成蛋白-2和碱性成纤维细胞生长因子对牙周膜细胞碱性磷酸酶活性的联合效应

目的 :了解重组人骨形成蛋白 - 2 (rhBMP - 2 )和碱性成纤维细胞生长因子 (bFGF)单独和联合作用对人牙周膜细胞 (PDLC)碱性磷酸酶 (ALP)活性的影响。方法 :体外培养人PDLC ,分别用不同浓度的rhBMP- 2和bFGF单独或联合作用 ,用酶动力学方法检测PDLC的ALP活性。结果 :5 0～ 2 0 0 μg/L浓度的rhBMP - 2可显著增强人PDLC的ALP活性 (P <0 .0 1) ,而 10 μg/L浓度的bFGF可显著抑制人PDLC的ALP活性(P <0 .0 1) ,rhBMP - 2和bFGF联合作用仍可较明显地增强人PDLC的ALP活性 (P <0 .0 5 )。结论 :rhBMP - 2和bFGF联合应用可增强人PDLC的ALP活性     

The effect of fibroblast growth factor-2 on the osteoinductive activity of recombinant human bone morphogenetic protein-2 in rat muscle

To clarify the effect of recombinant human basic fibroblast growth factor (FGF-2) on the osteoinductive activity of recombinant human bone morphogenetic protein-2 (BMP-2) in vivo, different amounts of FGF-2 (0, 16, 80 and 400 ng, and 2, 10 and 50 micro g: n=10 in each group), BMP-2 (2 micro g) and type I collagen as a carrier were mixed and implanted into rat calf muscles. Three weeks after implantation, compared with the controls, the radiopaque shadows of the implants were increased in the 16, 80 and 400 ng FGF-2-treated groups, but decreased in the 2, 10 and 50 micro g FGF-2-treated groups. In addition, alkaline phosphatase activity was increased in the 16, 80 and 400 ng FGF-2-treated groups but decreased in the 50 micro g FGF-2-treated group. Histological examination revealed increased bone formation in the 16, 80 and 400 ng FGF-2-treated groups. These results show that combined treatment with FGF-2 and BMP-2 has a biphasic effect on osteoinductive activity, i.e. it increases with low doses of FGF-2 and decreases with high doses of FGF-2.     

富血小板纤维蛋白膜(PRF)引导成骨能力的比较研究

目的:探讨及比较富血小板纤维蛋白膜(PRF)、海奥生物膜联合骨粉用于新西兰大白兔下颌骨缺损处早期引导骨生成的能力。方法:选取雌性新西兰大白兔9只,在其双侧下颌骨下缘上2mm做圆形骨缺损,A组植入骨粉后覆盖PRF,B组植入骨粉后覆盖海奥生物膜,C组植入骨粉。术后2,4,8周分期分组处死动物,取下颌骨缺损植骨区标本。标本行大体观察、CT及组织学观察成骨情况。结果:大体观察,骨缺损区早期骨愈合坚硬度A组>B组>C组。CT观察,骨缺损区骨密度早期A组>B组>C组,统计学分析,2周、4周时A组B组C组之间骨密度值分别具有统计学差异,8周组A组B组无统计学差异,与C组分别存在统计学差异。组织学观察,同期比较A组新生骨组织及再生血管化均优于B组、C组,新生骨排列较B组、C组规则。结论:PRF在骨缺损的修复中能够促进新骨的形成,提高新骨形成的时间和质量,对引导成骨起到有效促进作用。     

骨缝牵引中联合应用骨形态发生蛋白-2和骨保护素的实验研究

目的探索上颌骨在骨缝牵引时加入缓释骨形态发生蛋白-2（BMP-2）和骨保护素（OPG）对新骨形成的影响。方法以24只杂种犬为研究对象,随机分为A、B、C组。通过手术在上颌骨腭横缝植入自制的新型牵引器。A、C组术后5 d在牵引区附近注射缓释重组人骨形态发生蛋白-2/聚乳酸-羟基乙酸共聚物/纤维蛋白胶（rhBMP-2/PLGA/FS）,B、C组在牵引3周后注射人骨保护素/纤维蛋白胶（rhOPG/FS）。牵引1、2、4、6周后处死动物,采集标本进行组织学染色,并通过组织计量学方法检测腭横缝的组织改建情况。结果A、C组骨缝区成骨细胞功能活跃,透射电镜显示细胞内有大量高尔基复合体、线粒体及粗面内质网。牵引6周时,A、B、C组成骨细胞指数分别为38.5±7.7、35.7±6.5、41.7±11.0,破骨细胞指数分别为5.9±1.0、1.2±0.3、2.8±0.4,骨小梁厚度分别为（38.36±13.28）、（66.20±9.16）、（51.85±9.92）μm;B、C组表现出骨密度增加及破骨细胞指数下降。结论本实验所用牵引器能促进新骨生成;BMP-2与OPG在骨缝牵引过程中有协同作用,可以促进新骨形成及骨改建。     

Effect of prostaglandin E2 on recombinant human bone morphogenetic protein-2-stimulated osteoblastic differentiation in human periodontal ligament cells

Recombinant human (rh) bone morphogenetic protein-2 (BMP-2) stimulates osteoblastic differentiation in cells isolated from human periodontal ligament (HPLC), and this action of rhBMP-2 may be modulated by prostaglandins (PGs), which are local regulatory factors in the bone metabolism. In the present study, we investigated the effect of prostaglandin E2 (PGE2) on rhBMP-2-stimulated osteoblastic differentiation in cultured HPLC. rhBMP-2 (500 ng/ml)-stimulated alkaline phosphatase (ALPase) activity was enhanced by simultaneous treatment with low concentrations (10(-10)-10(-8) M) of PGE2, whereas a high concentration (10(-6) M) of PGE2 suppressed it. rhBMP-2 did not induce cyclo-oxygenase-2 (COX-2) mRNA expression or subsequent PGE2 production, whereas it remarkably suppressed rhIL-1 beta-induced COX-2 mRNA expression and PGE2 production. The rhBMP-2 action on osteoblastic differentiation in HPLC was also enhanced by co-treatment with 0.25 to 25 ng/ml of rh interleukin-1 beta (IL-1 beta). The ALPase activity stimulated by simultaneous treatment with rhBMP-2 and rhIL-1 beta was partially inhibited by addition of 10(-6) M of indomethacin, which completely inhibited rhIL-1 beta-induced PGE2 production. These results reveal that PGE2 at different concentrations exerts a biphasic effect on BMP-2-stimulated osteoblastic differentiation in HPLC, BMP-2 inhibits IL-1 beta-induced PGE2 production through suppressing COX-2 expression, and the BMP-2-stimulated osteoblastic differentiation may be enhanced by the endogenous PGE2 induced by BMP-2 and IL-1 beta. These suggest that BMP-2 action on osteoblastic differentiation in HPLC may be modulated by PGE2 in autocrine and paracrine fashions.