多孔载因子CPC促进骨缺损愈合的实验研究

[目的]探讨磷酸钙骨水泥复合rhBMP-2/明胶微球复合材料在治疗骨缺损时的降解、成骨性能。[方法]制备携载rhBMP-2的明胶微球(GMs),与磷酸钙骨水泥(CPC)复合,制备出rhBMP-2/GMs/CPC复合人工骨。取30只新西兰大白兔,在前臂桡骨中段制造人工骨缺损,随机分成3组,分别植入rhBMP-2/GMs/CPC/复合物(A组)、GMs/CPC(B组)、rhBMP-2/CPC(C组),术后6、12周分别进行X线检测、骨密度测定,术后12周处死动物,分别行生物力学测定,脱钙切片、HE染色,不脱钙切片进行荧光显微镜下观察双标间距,计算平均矿化率。[结果]与GMs/CPC、rhBMP-2/CPC组比较,复合材料植入后不同时间点的材料降解及成骨均高于对照组。12周A组标本生物力学实验测定结果表明指标接近正常,与B、C组比较有统计学差异。骨密度12周、新骨矿化率提示有统计学差异。[结论]rhBMP-2/GMs/CPC微球系统复合材料在体内易降解,具有良好成骨活性,是良好的骨修复材料。     

携载rhBMP-2微球的新型可注射自凝固复合人工骨的制备及特性研究

[目的]制备一种具有良好降解性和成骨活性、可注射的自凝固新型骨修复材料。[方法]制备携载rhBMP-2的聚乳酸与聚乙醇酸共聚物（PLGA）微球，并将其与rhBMP-2／磷酸钙骨水泥（CPC）复合，制备出rhBMP-2／PLGA微球／CPC复合人工骨。探讨了材料的特性，包括形貌、固化时间、抗压强度及反映材料体外降解速度的指标一体外降解液Ca、P浓度变化，测定复合材料rhBMP乏的释药速度及体外诱导MSCs细胞成骨分化的能力。[结果]与单纯CPC-rhBMP-2相比，复合材料的固化时间少量增加，抗压强度下降明显。体外降解速度及体外释药明显提高，释放的rhBMP-2具有骨诱导活性。[结论]rhBMP-2／PLGA微球／磷酸钙骨水泥新型复合人工骨是具有良好应用前景的骨修复材料。     

携载rhBMP-2微球的新型复合人工骨的释药及成骨活性研究

目的 制备一种具有良好降解性和成骨活性，可注射的自凝固新型骨修复材料。方法制备携载rhBMP-2的聚乳酸与聚乙醇酸共聚物（PLGA）微球，并将其与rhBMP-2／磷酸钙骨水泥（CPC）复合。制备出rhBMP-2／PLGA微球／CPC复合人工骨。测定了复合材料释药速度，将复合材料及rhBMP-2-CPC分别植入兔双侧股骨髁骨缺损区域，通过X线、组织学观察比较不同时期材料的降解及成骨情况。结果 复合材料各时间点的体外释药量均大于单纯rhBMP-2／CPC组．与单纯rhBMP-2／CPC材料相比较，复合材料植入体内后不同时间点材料的降解和新骨生成均高于单纯rhBMP-2／CPC植入组。结论 rhBMP-2／PLGA微球的掺入可明显加快rhBMP-2的释放。提高材料的降解速度和成骨活性。     

多孔磷酸钙人工骨与重组人骨形成蛋白2复合修复骨缺损的实验研究

目的研究多孔磷酸钙人工骨（porous calcium phosphate cement，PCPC）与重组人骨形成蛋白2（recombinant human bone morphogenetic protein2，rhBMP-2）复合后体外的缓释作用及其对兔骨缺损的修复作用。方法采用物理吸附法将rhBMP-2（0．4mg）溶液吸附至PCPC中，制备成PCPC／rhBMP-2复合材料。冻干后，扫描电镜观察复合材料内部形态。以包覆壳聚糖的PCPC／rhBMP-2为实验组，单纯PCPC／rhBMP-2为对照组，测试在模拟体液中的rhBMP-2缓释行为。取新西兰大白兔12只，股骨远端制成直径4．2mm，深5．0mm的骨缺损模型。将包覆壳聚糖的PCPC／rhBMP-2复合材料修复骨缺损作为实验组，以植入单纯PCPC作为对照组。术后观察动物一般情况，于4周和8周取材行X线片和组织学观察。结果扫描电镜显示PCPC／rhBMP-2复合材料孔隙中吸附了大量的rhBMP-2。rhBMP-2体外缓释：对照组rhBMP-2于150h基本全部释放；实验组rhBMP-2于350h缓释量约达99％，较对照组慢。动物实验：动物术后切口无感染，于4周行动自如。X线片示术后4周对照组骨缺损区材料清晰，实验组骨缺损区密度大部分接近宿主骨，材料模糊；8周对照组材料边缘较术后4周模糊，实验组骨缺损区密度已基本接近宿主骨。组织学观察，术后4周对照组可见少量成骨细胞和破骨细胞，实验组可见成熟骨组织和骨髓腔，新生骨逐渐取代材料；8周对照组可见大量成骨细胞和破骨细胞，少量新生骨并向材料内长入，实验组可见成熟骨小梁和骨髓组织。结论PCPC是rhBMP-2较理想的载体材料，复合后具有良好的诱导成骨作用，可作为一种新型复合人工骨修复骨缺损，具有良好的临床应用前景。     

复合rhBMP-2壳聚糖微球可注射磷酸钙骨水泥制备及成骨性能研究

[目的]研究复合重组人骨形态发生蛋白-2(rhBMP-2)壳聚糖微球的可注射磷酸钙骨水泥的成骨性能,从而构建一种具有骨传导和骨诱导性的新型可注射性植骨材料.[方法]制备负载rhBMP-2壳聚糖微球磷酸钙骨水泥材料,并将单纯磷酸钙骨水泥和单纯壳聚糖微球磷酸钙骨水泥材料作为对照.检测负载rhBMP-2壳聚糖微球骨水泥的rhBMP-2体外释药.在体外将青山羊骨髓干细胞(MSCs)与3种材料浸提液共同培养,通过检测碱性磷酸酶活性和骨钙素含量评价MSCs成骨分化.在体内,将3种材料植入青山羊骨缺损模型,通过组织学检测研究复合材料成骨能力.[结果]rhBMP-2在1 d时的释放量为12.6%,随后呈持续缓慢释放.在培养7、14 d,负载rhBMP-2壳聚糖微球磷酸钙骨水泥复合材料浸提液可显著提高MSCs的ALP活性和骨钙素含量,差异有统计学意义(P<0.05).组织学显示负载rhBMP-2壳聚糖微球磷酸钙骨水泥复合材料的新骨形成均优于其他组.[结论]复合rhBMP-2壳聚糖微球的可注射磷酸钙骨水泥复合材料是一种有应用前景的新型可注射骨移植材料.     

三种充填材料应用于山羊经皮椎体成形术的组织学评价

经皮椎体成形术（PVP）充填材料性能的优劣直接关系到PVP的成败，我们在山羊骨质疏松症（OP）模型上行PVP．充填聚甲基丙烯酸甲酯（PMMA）、可注射型的自固化磷酸钙人工骨（CPC）和复合重组人类骨形态发生蛋白-2（rhBMP-2）的CPC（rhBMP-2／CPC）三种材料。并行光镜观察，为寻找理想的PVP充填材料提供组织学依据。     

局部应用CPC/rhBMP-2复合材料对骨质疏松大鼠骨折愈合的影响

目的研究磷酸钙骨水泥（Calcium Phosphate Cement，CPC）与重组人骨形态发生蛋白-2（Recombinant Human Bone Morphogenefic Protein-2，rhBMP-2）复合材料对骨质疏松大鼠骨折愈合的影响。方法取4月龄雌性清洁大鼠106只，随机分成实验组96只，对照组10只。实验组行双侧卵巢切除术，对照组行假手术，术后行双能X线骨扫描确定骨质疏松动物模型形成。然后将实验组96只大鼠制作右侧股骨远端松质骨骨折，将上述动物随机分成3组，每组32只：（1）克氏针组，从髁间窝穿入直径1mm克氏针固定；（2）CPC组，将CPC从12号针头注入骨髓腔，手法固定至骨水泥凝固；（3）CPC-BMP组，将CPC与rhBMP-2复合材料用12号针头注入骨髓腔，手法固定至骨水泥凝固。术后第2、4、6、8wk三组各取8只动物处死，取手术侧股骨远端标本行压缩实验。结果在上述4个时间点，CPC-BMP组的骨折段最大载荷较克氏针组及CPC组增高（P〈0．05）；第2周CPC组骨折段最大载荷较克氏针组增高（P〈0．05）。结论（1）局部注射CPC-BMP复合材料可以促进骨质疏松大鼠骨折的愈合，提高骨折愈合后的强度；（2）CPC局部应用可以对骨折起到即时固定作用。     

新型可注射可降解磷酸钙骨水泥对成骨作用影响的实验研究

目的比较单纯的、添加锌锶等元素及复合rhBMP-2的新型可注射可降解磷酸钙骨水泥(CPC)植入机体后的降解及成骨作用,检验材料及其改进剂型在临床应用的可能性。方法将24只雄性新西兰白兔随机分为A,B,C三组,分别为9、9和6只,制备双侧胫骨结节骨缺损模型；于骨缺损处分别置入：A组为单纯新型可注射可降解CPC材料,B组为含锌锶等微量元素的该CPC材料,C组为该CPC材料与rhBMP-2的复合物。观察其全身及植入局部反应,术后4、8、16周取材CT扫描、肉眼及光学显微镜观察材料植入后的局部反应情况并对比观察该三种材料的降解及骨生长情况。组织切片摄像后,每组每时间点各取5幅相片用软件进行图文处理,求得术后骨组织切片中骨组织含量百分比。结果CT扫描、肉眼及光学显微镜观察发现单纯的和含锌锶等微量元素的CPC材料可引导新骨形成,但新骨形成少且慢,与材料降解不同步；复合rhBMP-2的CPC材料组新生骨形成多而早,基本与材料降解同步。术后骨组织含量百分比测定,A组为(41．7±16．6)%,B组为(31．2±12．2)%,C组为(71．7±21．0)%,复合BMP的CPC材料组术后骨组织切片中骨组织含量显著高于另外两组(P＜0．01)。结论复合rhBMP-2的可注射可降解CPC材料可较早地诱导新生骨生成,与单纯的及含锌锶等微量元素的可注射可降解CPC材料相比,更适合于临床使用。     

磷酸钙骨水泥复合物/脐带间充质干细胞凝胶构建组织工程骨

目的构建新型可注射强化型磷酸钙骨水泥复合物/脐带间充质干细胞凝胶组织工程骨,探讨其力学性能,细胞活性和成骨作用。方法选用第四代hUCMSCs,1.2%海藻酸钠水凝胶构建hUCMSCs水凝胶微球。高温煅烧钙/磷比约为1.9的磷酸氢钙和碳酸钙混合物,按摩尔质量比以1：1混合制备CPC粉末。15%Chitosan,8mm长度可吸收纤维用于提高CPC复合物力学强度。实验组分为四组：（1）单纯hUCMSCs微球;（2）CPC＋hUCMSCs微球;（3）CPC＋chitosan＋hUCMSCs微球;（4）CPC＋chitosan＋可吸收纤维＋hUCMSCs微球,分别检测力学性能,细胞活性和成骨作用。结果新型组织工程骨力学性能显著增强,抗弯曲强度提高到（11.7±2.1）MPa,弹性模量提高到（2.0±0.4）GPa,断裂功提高到（1.65±0.66）kj/m2（P〈0.05）。hUCMSCs的ALP活性第7天时明显增高,第14天时达峰,第21天时有所减弱,各组间比较无明显统计学差异（P〉0.1）。第7天时,所有实验组中的hUCMSCs均见少量矿物合成。第14天和第21天时,矿物合成数量明显增多。hUCMSCs中ALP基因表达培养第1天最低,第4天达峰,第8天时稍减弱。OC基因表达第8天达峰。结论构建完成新型可注射强化型磷酸钙骨水泥/脐带间充质干细胞凝胶构建组织工程骨。水凝胶微球中hUCMSCs在CPC中具有良好的成骨作用。CPC-chitosan-可吸收纤维组织工程骨力学性能满足松质骨力学要求,支持水凝胶微球中hUCMSCs的细胞活性和成骨作用。为组织工程骨研究和临床应用提供新思路和新方法。     

多孔明胶微球/磷酸钙骨水泥的制备及其性能研究

[目的]制备多孔明胶微球/磷酸钙骨水泥,并于体内体外研究其各种性能.[方法]双相乳化冷凝聚合法制备明胶微球,以不同比例与磷酸钙骨水泥复合(0%,2.5%,5%),制备多孔磷酸钙骨水泥,测定材料孔径率及抗压强度,筛选出最佳比例.消化法培养成骨细胞接种于常规及多孔磷酸钙骨水泥支架上,扫描电镜观察细胞形态;不同材料浸提液(0%,2.5%明胶微球/磷酸钙骨水泥及聚苯乙烯)分别与成骨细胞共培养,以MTT法测定细胞增殖率,试剂盒检测碱性磷酸酶水平.将磷酸钙骨水泥及2.5%明胶微球/磷酸钙骨水泥分别植入山羊椎体内,6个月后收集标本,分别进行X线影像学及组织学观察,评估其降解情况.[结果]不同比例明胶微球/磷酸钙骨水泥的总孔径率、大孔率及抗压强度分别为:38.7%、0%、12.1 MPa(0%);67.5%、40.6%、8.0 MPa(2.5%);72.2%、45.6%、5.0 MPa(5%).成骨细胞在明胶微球/磷酸钙骨水泥上生长良好,细胞增殖及碱性磷酸酶水平均明显高于单纯磷酸钙骨水泥组,与聚苯乙烯组未见明显差异.多孔明胶微球/磷酸钙骨水泥6个月后在体内大部分已降解,而磷酸钙骨水泥未见明显降解.[结论]复合明胶微球可显著提高磷酸钙骨水泥的孔径率促进其降解,增加其生物活性,这种多孔磷酸钙骨水泥可作为非负重部位的骨替代物.     

Controlled‐release of bone morphogenetic protein‐2 from a microsphere coating applied to acid‐etched Ti6AL4V implants increases biological bone growth in vivo

A central clinical challenge regarding the surgical treatment of bone and joint conditions is the eventual loosening of an orthopedic implant as a result of insufficient bone ingrowth at the bone–implant interface. We investigated the in vivo effectiveness of a coating containing recombinant human bone morphogenetic protein‐2 (rhBMP‐2)‐loaded microspheres applied to acid‐etched Ti6Al4V cylinders for implantation. Three groups of rabbits (24 per group) were used for implantation: (1) acid‐etched Ti6Al4V implants coated with a mixture of rhBMP‐2‐loaded microspheres (125 ng rhBMP‐2/mg microspheres) and α‐butyl cyanoacrylate; (2) acid‐etched, uncoated implants; and (3) bare, smooth uncoated implants. After implantation, 12 rabbits from each group were used for bone ingrowth determination at 4, 5, 6, 7, 8, and 12 weeks (2 rabbits per time point), while the remainder were used for histological analysis and push‐out testing at 12 weeks. Scanning electron microscopy showed significant improvement in bone growth of the rhBMP‐2 microspheres/α‐butyl cyanoacrylate group compared with the other groups (p < 0.01). Histological analysis and push‐out testing also demonstrated enhanced bone growth of the rhBMP‐2 group over that in the other two groups (p < 0.01). The rhBMP‐2 group showed the most significant bone growth, suggesting that coating acid‐etched implants with a mixture of rhBMP‐2‐loaded microspheres and α‐butyl cyanoacrylate may be an effective method to improve the osseointegration of orthopedic implants. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:744–751, 2014.     

The use of an injectable, biodegradable calcium phosphate bone substitute for the prophylactic augmentation of osteoporotic vertebrae and the management of vertebral compression fractures.

STUDY DESIGN: A biomechanical study comparing two materials for augmentation of osteoporotic vertebral bodies and vertebral bodies after compression fracture. OBJECTIVES: To compare an injected, biodegradable calcium phosphate bone substitute with injected polymethylmethacrylate bone cement for strengthening osteoporotic vertebral bodies and improving the integrity of vertebral compression fractures. SUMMARY OF BACKGROUND DATA: Injection of polymethylmethacrylate bone cement into fractured vertebral bodies has been used clinically. However, there is concern about thermal damage to the neural elements during polymerization of the polymethylmethacrylate bone cement as well as its negative effects on bone remodeling. Biodegradable calcium phosphate bone substitutes have been studied for enhancement of fixation in fractured vertebrae. METHODS: Forty fresh osteoporotic thoracolumbar vertebrae were used for two separate parts of this study: 1) injection into osteoporotic vertebrae: intact control (n = 8), calcium phosphate (n = 8), and polymethylmethacrylate bone cement (n = 8) groups. Each specimen then was loaded in anterior compression until failure; 2) injection into postfractured vertebrae: calcium phosphate (n = 8) and polymethylmethacrylate bone cement (n = 8) groups. Before and after injection, the specimens were radiographed in the lateral projection to determine changes in vertebral body height and then loaded to failure in anterior bending. RESULTS: For intact osteoporotic vertebrae, the average fracture strength was 527 +/- 43 N (stiffness, 84 +/- 11 N/mm), 1063 +/- 127 N (stiffness, 157 +/- 21 N/mm) for the group injected with calcium phosphate, and 1036 +/- 100 N (stiffness, 156 +/- 8 N/mm) for the group injected with polymethylmethacrylate bone cement. The fracture strength and stiffness in the calcium phosphate bone substitute group and those in the polymethylmethacrylate bone cement group were similar and significantly stronger than those in intact control group (P < 0.05). For the compression fracture study, anterior vertebral height was increased 58.5 +/- 4.6% in the group injected with calcium phosphate and 58.0 +/- 6.5% in the group injected with polymethylmethacrylate bone cement as compared with preinjection fracture heights. No significant difference between the two groups was found in anterior vertebral height, fracture strength, or stiffness. CONCLUSION: This study demonstrated that the injection of a biodegradable calcium phosphate bone substitute to strengthen osteoporotic vertebral bodies or improve vertebral compression fractures might provide an alternative to the use of polymethylmethacrylate bone cement.     

无定形磷酸钙负载rhBMP-2纳米缓释体的生物相容性及安全性

[目的] 观察无定形硫酸钙(ACP)负载重组人骨形态发生蛋白-2(rhBMP-2/ACP)纳米缓释体的生物相容性及安全性,为新型材料的临床应用提供必要的实验依据.[方法] 对材料分别进行体外溶血实验、微核实验、急性毒性试验、热原试验、皮内刺激反应实验和短期肌内埋置试验、骨内植入实验.[结果] 材料浸提液对兔血无溶血现象,对小鼠骨髓细胞无遗传毒性作用,未引起小鼠急性毒性反应、兔热原反应、兔皮内刺激反应;兔短期肌肉埋置、骨内植入后局部无明显炎症反应、无组织坏死,材料逐渐发生降解,能与组织有机融合.[结论] rhBMP-2/ACP纳米缓释体的生物相容性及安全性符合IS010993、GB/T16886规定的体内植入物的生物学评价标准,可作为骨修复替代材料用于相关疾病的临床治疗.     

珊瑚复合人工骨骨诱导活性的实验研究

目的弥补单纯珊瑚无骨诱导活性、骨修复能力弱等缺陷,为临床提供理想的骨移植替代材料。方法将珊瑚与胶原和重组人骨形成蛋白２（ｒｈＢＭＰ２）复合,制备珊瑚／胶原／ｒｈＢＭＰ２复合人工骨,将其植入大鼠背部肌肉陷窝,以珊瑚／胶原和珊瑚／ｒｈＢＭＰ２复合人工骨植入作对照;取材后通过组织学方法和图像分析评价其骨诱导活性。结果珊瑚／胶原／ｒｈＢＭＰ２植入后１周,在局部诱导出软骨细胞分化和软骨基质形成;植入后４周,形成含骨髓的板层骨;诱导成骨的量有明显的ｒｈＢＭＰ２剂量依赖性（Ｐ＜０．０１）;而珊瑚／胶原和珊瑚／ｒｈＢＭＰ２植入区均无骨或软骨形成。结论珊瑚／胶原／ｒｈＢＭＰ２复合人工骨具有良好的异位诱导成骨活性,是一种较理想的骨移植替代材料。     

Effect of rhBMP‐2 on tibial plateau fractures in a canine model

This study was to determine the efficacy of recombinant human bone morphogenetic protien‐2 (rhBMP‐2)/calcium phosphate matrix (CPX) paste to accelerate healing in a canine articular fracture model with associated subchondral defect. rhBMP‐2/CPX (BMP), CPX alone (CPX) or autogenous bone graft (ABG) was administered to a canine articular tibial plateau osteotomy with a subchondral defect in each of 21 female dogs. The unoperated contralateral limbs served as controls. Ground reaction forces, synovial fluid, radiographic changes, mechanical testing, bone density, and histology of bone and synovium were analyzed at 6 weeks after surgery. Radiographic analysis demonstrated that the BMP and CPX groups showed improved bony healing compared to the ABG group at week 6. Histomorphometric analysis demonstrated that the BMP group had significantly increased trabecular bone volume compared to the CPX and ABG groups. Mechanical testing revealed that the BMP group had significantly greater maximum failure loads than the ABG group. Histological analysis demonstrated that the BMP group had significantly less sub‐synovial inflammation than CPX group. This study demonstrated that rhBMP‐2/CPX accelerated healing of articular fractures with subchondral defect compared to ABG in most of the parameters evaluated, and had less subsynovial inflammation than the CPX alone in a canine model. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 466–471, 2009     

Calcium Phosphate Cement with BMP-2-loaded Gelatin Microspheres Enhances Bone Healing in Osteoporosis: A Pilot Study

Background

The capacity for bone healing reportedly is limited in osteoporosis with a less than ideal environment for healing of bone grafts. We therefore developed a composite bone substitute with rhBMP-2 loaded gelatin microsphere (GM) and calcium phosphate cement (CPC) to use in osteoporosis.

Questions/purposes

We asked whether (1) controlled release of rhBMP-2 could be improved in this composite bone substitute and (2) increasing factors released from the bone substitute could accelerate osteoporotic bone healing.

Methods

We soaked rhBMP-2/GM/CPC and rhBMP-2/CPC composites in simulated body fluid for 28 days and then determined the amount of rhBMP-2 released. Both composites were implanted in bone defects of osteoporotic goats and left in place for 45 and 140 days; the specimens then were evaluated mechanically (pushout test) and morphologically (CT scanning, histology).

Results

The in vitro study showed the new composite released more rhBMP-2 compared with rhBMP-2/CPC. CT showed the defects healed more quickly with new grafts. The bone mineralization rate was greater in rhBMP-2/GM/CPC than in rhBMP-2/CPC after 45 days of implantation and the pushout test was stronger after 45 and 140 days of implantation.

Conclusions

The new graft composite released more loaded factors and appeared to repair osteoporotic bone defects.

Clinical Relevance

These preliminary data suggest the new composite can be used as a bone substitute to accelerate healing of fractures and bone defects in osteoporosis.     

Effect of rhBMP‐2 and VEGF in a vascularized bone allotransplant experimental model based on surgical neoangiogenesis

We have demonstrated survival of living allogeneic bone without long‐term immunosuppression using short‐term immunosuppression and simultaneous creation of an autogenous neoagiogenic circulation. In this study, bone morphogenic protein‐2 (rhBMP‐2), and/or vascular endothelial growth factor (VEGF), were used to augment this process. Femoral diaphyseal bone was transplanted heterotopically from 46 Dark Agouti to 46 Lewis rats. Microvascular repair of the allotransplant nutrient pedicle was combined with intra‐medullary implantation of an autogenous saphenous arteriovenous (AV) bundle and biodegradable microspheres containing buffer (control), rhBMP‐2 or rhBMP‐2 + VEGF. FK‐506 given daily for 14 days maintained nutrient pedicle flow during angiogenesis. After an 18 weeks survival period, we measured angiogenesis (capillary density) from the AV bundle and cortical bone blood flow. Both measures were greater in the combined (rhBMP‐2 + VEGF) group than rhBMP‐2 and control groups (p < 0.05). Osteoblast counts were also higher in the rhBMP‐2 + VEGF group (p < 0.05). A trend towards greater bone formation was seen in both rhBMP2 + VGF and rhBMP2 groups as compared to controls (p = 0.059). Local administration of VEGF and rhBMP‐2 augments angiogenesis, osteoblastic activity and bone blood flow from implanted blood vessels of donor origin in vascularized bone allografts. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 561–566, 2013     

多孔双相陶瓷复合重组人骨形态发生蛋白-2和碱性成纤维细胞生长因子缓释微球的异位成骨活性

目的 观察复合重组人骨形态发生蛋白-2(rhBMP-2)和碱性成纤维细胞生长因子(bFGF)缓释微球的多孔双相陶瓷(BCP)异位成骨活性.方法 A组(rhBMP-2+bFGF缓释微球/BCP)、B组(BCP)、C组(bFGF缓释微球/BCP)、D组(rhBMP-2缓释微球/BCP),其中bFGF和rhBMP的浓度各为5、15μg.规格均为3mm×8mm×28mm.将材料植入兔背部肌袋内,术后4、8、12周分别取材,行大体、组织学观察,测量新骨面积和血管生成.结果 8、12周A组成骨活性较其他组优,差异有统计学意义 (P<0.05).A组4周材料中有较多间充质细胞,并有少量毛细血管的生长;8周时可见散在分布的不成熟新生骨组织,材料基本降解;12周时出现成熟度较低的编织骨,为膜内成骨.结论 BCP复合rhBMP-2和bFGF缓释微球具有良好的异位成骨活性.

Abstract：

Objective To investigate the heterotopic bone formation ability of biphasic ceramics phosphate (BCP) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF) microspheres.Methods The rabbits were divided into group A (rhBMP-2+bFGF/BCP), group B (BCP), group C (bFGF/BCP), and group D (rhBMP-2/BCP). The concentrations of bFGF and rhBMP were 5 μg and 15 μg, respectively. All the samples were 3 mm×8 mm×28 mm in size. The muscle pouches of the rabbits were implanted with the samples. The ectopic bone formation was evaluated in the following aspects: histology, osteogenic area, and blood capillary number at 4th, 8th and 12th week after operation.Results At any specified time point, the value of the heterotopic bone formation was significantly higher in group A than other groups (P< 0.05). The histological analysis showed that group A had more mesenchymal (MES) cells and fewer blood capillaries at 4th week after operation. At 8th week, the composite was degradated and diffusely distributed, and the immature new bone was found. There were low-grade mature woven bones at 12th week, and the membranous bone formation occurred.Conclusion BCP combined with rhBMP-2 and bFGF microspheres has good heterotopic bone formation ability.     

载骨髓间充质干细胞的磷酸钙骨水泥的制备

[目的]观察载骨髓间充质干细胞(Bone marrow mesenchymal stem cells,MSCs)明胶微球磷酸钙骨水泥(Calcium phosphate cement,CPC)的理化性质,探究新型CPC的制备。[方法]制备载MSCs的明胶微球后,分别以0%(A),2.5%(B),5%(C),10%(D)的质量比(w/w)与CPC复合。测定其抗压强度、初凝时间、孔隙率和大孔率等理化性质,进一步检测MSCs在CPC中的成长情况。[结果]在各组CPC中MSCs均生长良好,且与空白对照组比较,随着明胶微球比例的增大,各组CPC的抗压强度逐步降低,初凝时间延长,孔隙率增大,大孔率增高(P<0.05)。[结论]载MSCs的明胶微球与CPC的质量比为5%时,CPC的理化性质最为理想,细胞生长良好,此结果为下一步体内实验奠定了坚实的基础。