应用猪主动脉去细胞瓣膜支架体外构建组织工程心脏瓣膜

目的　探讨应用猪去细胞瓣膜支架体外构建组织工程心脏瓣膜的可行性。　方法　采用去垢剂、渗透压改变和核酸酶消化的方法制备猪主动脉瓣去细胞瓣膜支架 (实验组 ) ,用去内皮细胞主动脉瓣作对照 (对照组 ) ,并对两组含水量、可溶性蛋白含量、热皱缩温度、力学性能和组织相容性进行测定。培养犬主动脉壁间质细胞和内皮细胞 ,将其种植于实验组去细胞支架上 ,观察细胞生长情况 ,并测定内皮细胞合成前列环素的功能。　结果　实验组瓣膜细胞成分完全从瓣膜中去除 ,与对照组新鲜瓣膜相比 ,含水量增高 (P<0 .0 5 ) ,可溶性蛋白含量减少 (P<0 .0 5 ) ,热皱缩温度和抗张强度无明显变化。实验组瓣膜组织相容性试验显示 ,材料组织相容性好 ,体内降解时间为 10周 ;犬主动脉壁间质细胞和内皮细胞在瓣膜表面生长良好 ,内皮细胞具有合成分泌前列环素的功能。　结论　采用去垢剂、渗透压改变和核酸酶消化的方法制备猪主动脉瓣去细胞瓣膜支架 ,在去除细胞和可溶性蛋白质的同时保持了瓣叶的基本结构和力学性能 ;以其为支架体外构建组织工程心脏瓣膜的细胞不仅能在材料表面生长 ,还能合成、分泌血管活性物质 ,是具有生理功能的组织工程心脏瓣膜。

In Vitro Experience with Tissue Engineered Heart Valve on Decellular Porcine Aortic Valve

ObjectiveTo evaluate the feasibility of tissue engineered heart valve on decellular porcine aortic valves.MethodsA detergent and enzymatic extraction process has been developed to remove cellular components from porcine aortic valve(experimental group), de-endothelial porcine aortic valve (control group) as comparison.Shrinkage temperature, tensile stress and the content of hydrogen, soluble protein were detected in the experimental group and control group. Cells of canine aortic wall and aortic endothelium were seeded on the decellular valves in experimental group. The seeded cells' growth condition were observed and their function of prostaglandin I 2 (PGI 2) synthesis were detected.ResultsCellular components were completely removed from porcine aortic valves in experimental group. Hydrogen were significantly increased (P<0.05) and soluble protein were significantly decreased (P<0.05) in experimental group. Decellular valve had good biocompatibility and was completely degradable at the end of the 10th week. Canine aortic wall interstitial cells and aortic endothelium grew well on the decellular valves. The seeded endothelium can synthesize and secret PGI 2 .ConclusionsCellular components and soluble protein are removed from porcine aortic valves with the detergent and enzymatic extraction process, meanwhile the collagen structure and mechanic characteristics of the decellular valves are well preserved. The seeded cells grow well on the decellular valves and synthesize, and secret angio-active substances, which confirm the tissue engineered heart valve constructed with the decellular valves has physiological function.