可吸收生物材料聚羟基乙酸与滑膜细胞共同构建组织工程化腱鞘

Construction of tissue engineered tendon sheath with polyglycolic acid and synoviocyte

目的:目前对于腱鞘的修复主要运用网腱膜、自体静脉和生物膜等腱鞘的替代品,其缺点是无生物学功能,不能达到真正意义上的结构与功能的重建。实验采用可吸收生物材料聚羟基乙酸与滑膜细胞共同构建组织工程化腱鞘,验证其可行性。方法:实验于2005-09/2007-03在上海市第九人民医院组织工程实验室完成。取Leghorn鸡滑膜腱鞘,体外消化培养获得滑膜细胞并大量扩增后,取第2代滑膜细胞与聚羟基乙酸生物材料复合,形成大小1.5cm×1.0cm,厚1.0mm左右的细胞生物学支架,体外培养6d后,将支架包绕在硅胶管上回植到裸鼠皮下及将支架直接回植于鸡爪原位腱鞘缺损处,以未接种细胞的单纯聚羟基乙酸包绕硅胶管为对照组,体内培养3,6周后取材观察。整个实验过程中对滑膜细胞及两种情况下构建形成的组织工程化腱鞘进行形态学和组织学观察。结果:①所获得的滑膜细胞分为A、B型滑膜细胞,A型滑膜细胞形态呈巨噬细胞样,B型滑膜细胞形态呈成纤维样,且随着滑膜细胞的扩增,A型滑膜细胞比例逐渐减少,到第2代时,基本全部为B型细胞。②在裸鼠皮下构建的组织工程化腱鞘第3周、第6周时有新生组织产生,且具有弹性,失去支撑后仍可保持形状,而单纯聚羟基乙酸对照组在第3周时只有少量新生组织,到第6周时新生组织消失。③在Leghorn鸡原位处构建的组织工程化腱鞘第3周时内壁光滑,与肌腱无粘连,第6周时形成与正常腱鞘相似鞘膜。结论:应用组织工程技术可以构建与正常组织相似的组织工程腱鞘。

AIM： Currently, succedaneum is used for repairing tendon sheath such as net tendinous membrane, autoallergic vein and biomembrane. It is disadvantage of no biological function, and it cannot fully reconstruct the structure and function. This study was designed to study the feasibility of engineered tendon sheath using synoviocytes and polyglycolic acid. METHODS： The experiment was performed at the Laboratory of Tissue Engineering, Shanghai Ninth People＇s Hospital from September 2005 to March 2007. Synoviocytes were collected from Leghorn chicken synovium tendon sheath, and expanded in vitro. The 2^nd passage synoviocytes were combined with polyglycolic acid to form 1.5 ×1.0 ×0.1 cm^3 cytobiological stent. Six days later, the silica gel tube encased with stent was replanted in athymic mouse subcutaneously and the stent was replanted directly in defected epitenon of chicken claw. Silica gel tube only encased with polyglycolic acid was considered as control group. The synoviocytes and endpieces of the complexes were harvested at weeks 3 and 6 for histological and morphological analysis. RESULTS：①The synoviacytes were divided into two groups, type A cells （macrophage-like synoviacytes） and type B cells （fibroblast-like synoviacytes）. During the period of the cells＇ amplification, the type A cells＇ ratio became less, and it was almost the type B cells in the 2^nd passage cells. ②At the 3^rd and 6^th weeks there was some tissue neogenesis in the subcutaneous complexes implanted into the athymic mouse, and the tissue was flexible and retained its original shape, while the control group with only polyglycolic acid replanted had just little tissue neogenesis at the 3^nd week and the tissue disappeared at the 6^th week. ③At the 3^nd week, the complexes implanted into the chicken＇s claws had smooth surfaces along the inner wall of the membrane .sheath and did not show signs of tendon adhesion, and it formed the tissue like normal at the 6^th week. CONCLUSION： It is quite possible to construct a tissue engineered tendon sheath that is similar to the natural one.