缓释rhVEGF的PDLLA/nHA复合多孔支架制备及血管诱导活性研究  

Fabrication of rhVEGF releasing poly-DL-lactic acid/nano-hydroxyapatite porous scaffold and its vessel inductive activity

作　　者：雷鸣[1] 杨述华[1] 迮仁浩[1] 王友法[2] 田建宁[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,武汉430022 [2]武汉理工大学生物材料与工程研究中心,430070

出　　处：《国际骨科学杂志》2010年第2期121-124,共4页International Journal of Orthopaedics

摘　　要：目的研究制备消旋聚乳酸/纳米羟基磷灰石(PDLLA/nHA)多孔支架复合并缓释重组人血管内皮细胞生长因子(rhVEGF165),并观察其诱导血管形成的生物特性,为进一步实验提供理论依据。方法将rhVEGF165与PDLLA溶液物理混匀,用冷冻相诱导分离法制备PDLLA/nHA复合多孔支架,制成直径3 mm、高8～10 mm圆柱状支架。电镜观察其结构特点,检测在缓释rhVEGF的PDLLA/nHA支架降解过程中rhVEGF释放量,绘制释放曲线。分离培养兔骨髓间充质干细胞(BMSC),接种于复合和未复合rhVEGF的PDLLA/nHA支架。将细胞支架复合物植入兔大腿肌袋中,4周后取出材料行vWF免疫组化染色,观察新生血管形成情况。结果每个缓释rhVEGF的PDLLA/nHA支架含rhVEGF 4μg,材料孔径为20～100μm,孔隙率约90%,孔径联通性好。免疫组化及新生血管密度检测显示缓释rhVEGF支架+BMSC(rhVEGF-PDLLA/nHA+BMSC,VPB组)新生血管密度较空白支架(PDLLA/nHA,P组)之差异非常显著(P<0.001),较空白支架+BMSC(PDLLA/nHA+BMSC,PB组)之差异显著(P<0.05)。结论缓释rhVEGF的PDLLA/nHA多孔支架的结构特点适合细胞生长,有较强的血管诱导活性,是血管化组织工程的理想材料。Objective To investigate the manufacture of a rhVEGF releasing Poly-DL-lactic acid （PDLLA）/nano hydroxyapatite（nHA） porous scaffold and its vessel inductive activity in rabbit. Methods We were blended rhVEGFI65 and PDLLA solution, then fabricated the rhVEGF releasing PDLLA/nHA porous scaffold by a cryogenic induced phase separation method, every scaffold was made a cylindrical shape with a diameter of 3 mm and a height of 8 to 10 mm. We were observed the uhrastrueture characters by scanning electronic microscope, detected the releasing amount during the degradation of PDLLA/nHA scaffold, isolated and cultured rabbit bone marrow mesenchymal stem cells（BMSC）, seeded cells in rhVEGF composited and non -composited PDLLA/nHA scaffolds, and implanted the cell/scaffold composite into rabbit＇s thigh intramuscularly for 4 weeks, then took out the composites and took a vWF immunohistochemical stain to observe the neovascularization. We divided rabbits into 3 groups： rhVEGF releasing PDLLA/nHA with BMSC group（VPB group）, PDLLA/nHA with BMSC group（PB group） and blank control group（P group）. Results Every rhVEGF releasing PDLLA/nHA porous scaffold had a good pores connectivity with a pore diameter of 10- 100um, and contained about 4 ug rhVEGF. The new vessels＇ density and diameter were significantly greater in VPB group than in P group （P〈0. 001 ）, and were greater than in PB group （P〈0. 05）, but showed no significant difference between P and PB group （P〉0. 05）. Conclusion The rhVEGF releasing PDLLA/nHA porous scaffold fabricated by using cryogenic induced phase separation method has a good biocompatibility and vascular inducibility which was a ideal tissue engineering scaffold in vascularization.

关 键 词：rhVEGF 多孔支架 BMSC PDLLA/nHA 血管诱导 

分 类 号：R318.08[医药卫生—生物医学工程]