显微外科技术预构组织工程骨血管化的比较

A comparative study on vascularization of three types of prefabricated pedicled tissue engineered bone

目的比较3种带血管蒂组织瓣预构组织工程骨血管化的效果，探讨其影响血管化的方式。方法设计以膝最下血管为蒂的股骨骨膜瓣包裹、贯穿，腹壁浅血管为蒂的筋膜瓣包裹体外培养的磷酸三钙（β—TCP）＋骨髓间充质干细胞（MSCs）复合物等3种组织工程预构骨动物模型，以β-TCP＋MSCs复合物和单纯β-TCP皮下埋放为对照，通过墨染透明标本、墨染苏木素-伊红（HE）切片、第8因子相关抗原免疫组织化学染色、铸型标本扫描电镜观察等指标，分析带血管蒂组织瓣对组织工程预构骨血管化的影响。结果新生血管自组织瓣与材料结合的界面向材料内部生长，组织瓣处理各组的血管化指标均显著优于非组织瓣处理组（P〈0．05），前者中血管化的优劣递次为A、B、C组（P〈0．05）；材料内部的孔隙中亦可观察到散在的血管，复合了MSCs的D组血管化优于单纯材料的E组（P〈0．01）。结论影响带血管蒂组织瓣组织工程预构骨血管化的因素有：（1）与材料复合的组织瓣血供及其与材料接触面积的大小；（2）提供给材料内部促进血管发生的细胞及细胞因子的种类及密度（包括内源性和外源性的）；（3）材料本身的特性，如组织相容性、是否有贯通的微孔等。

Objective To compare the vascularized results of three prefabricated pedicled tissue engineered bone and explore how the pedieled flaps affect the vaseularization of prefabricated bone. Methods The compound β-TCP ＋ MSCs was in vivo treated with 3 pedieled flaps, the first one was enveloped femoral periosteal flap pedieled with supreme genual vessels ,the second one was transfixed femoral periosteal flap pedieled with supreme genual vessels, the third one was enveloped faseial flap pedieled with su pertleial epigastrie vessels. Thel^-TCP ＋ MSCs and [^-TCP subcutaneously embedded were set as controlled groups. The vaseularization was assessed by ink perfusing transparent specimen, HE histological observation, Ⅷ factor related antigen immunohistochemical stain and SEM at time intervals of 2,4,8,16 weeks after operation. Results Two origins of new vessels were observed in the prefabrleated tissue engineered bone,one came from the interface of β-TCP and flap,the other dispersed over the tiny pores of β-TCP. The vaseularization of flap-treated groups （ A, B, C ） was better than non-flap-treated group （D） （ P 〈 0.05 ）, and the decreasing order was A 〉 B 〉 C （ P 〈 0.05 ）. The group of β-TCP ＋ MSCs was better in vaseularization than simple β-TCP（P 〈 0.05 ）. Conclusion The factors that affect the vaseularization of prefabricated pedieled bone could be considered as follows： （ 1 ） the area that the flap contacts with the material; （2） the categories and density of cells and eytokines which transfer to the inner pores of material to generate and induct new vessels ; （ 3 ） the characteristics of material itself, which include biocompatibility, interconnected pores, etc.