摘要
目的 通过组织工程技术方法 ,研究细胞 -材料复合体体内骨再生的能力。 方法 采用材料学自组装技术的原理 ,以 I型胶原蛋白为分子模板 ,引导钙磷盐在液相中的矿化 ,制备具有天然骨基质层状结构的羟基磷灰石 /胶原复合材料 ,并以热致分相法制备羟基磷灰石 /胶原 -聚乳酸 [hydroxyapatite/collagen- poly- (L- lactic acid) ,HAC-PL A]复合三维多孔框架 ,与重组人骨形成蛋白 2 (recombinant human bone morphogenetic protein2 ,rh BMP- 2 )诱导分化后的兔骨膜细胞构建组织工程骨 ,进行体外电镜及组织学观察。将 3月龄裸鼠 18只 ,分为 3组 ,每组 6只。A组皮下注射经 rh BMP- 2处理后的骨膜细胞悬液 ,B组植入未复合细胞的 HAC- PL A,C组植入 rh BMP- 2处理后的细胞 -材料复合体。术后 8周取材行组织学观察 ,C组与 B组及 A组进行比较。 结果 三维多孔 HAC- PL A框架材料孔隙率大于 90 % ,孔径为 5 0～ 30 0 μm。骨膜细胞经 5 0 0 ng/ml的 rh BMP- 2处理后 ,与改善亲水性后的 HAC- PL A三维多孔框架复合 ,构建组织工程骨。电镜显示接种的细胞能在此组织工程骨内部生长增殖 ,术后 8周 C组有新骨形成 ,A组注射部位表面平整 ,无新生物 ,B组为纤维组织 ,无骨及软骨形成。 结论 所构建的组织工程骨有望成?
Objective To investigate bone regeneration of the cell biomaterial complex using strategies of tissue engineering based on cells. Methods Hydroxyapatite/collagen (HAC) sandwich composite was produced to mimic the natural extracellular matrix of bone, with type Ⅰ collagen serving as a template for apatite formation. A three dimensional ploy porous scaffold was developed by mixing HAC with poly(L lactic acid) (PLA) using a thermally induced phase separation technique (TIPS). The rabbit periosteal cells were treated with 500 ng/ml of recombinant human bone morphogenetic protein 2(rhBMP 2), followed by seeded into prea2wet HACa2PLA scaffolds. Eighteen a(c)3 month nude mice were implanted subcutaneously cell suspension (group A, ?n?=6), simple HAC PLA scaffold (group B, ?n?=6) and cell biomaterial complex(group C, ?n?=6) respectively. Results Using type I collagen to template mineralization of calcium and phosphate in solution, we get HAC sandwich composite, mimicking the natural bone both in composition and microstructure. The three dimensional HAC PLA scaffold synthesized by TIPS had high porosity up to 90%, with pore size ranging from 50 μm to 300 μm. SEM examination proved that the scaffold supported the adhesion and proliferation of the periosteal cells. Histology results showed new bone formation 8 weeks after implantation in group C. The surface of group A was smooth without neoplasma. Fibrous tissue invasion occured in group B and no bone and cartilage formations were observed. Conclusion The constructed tissue engineering bone has emerged as another promising alternative for bone repair. ?3/4
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2005年第2期100-104,共5页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家高技术研究发展计划资助项目 (863) (71 5- 0 0 9-0 1 50 )~~
