羊膜细胞Ⅰ型胶原蛋白支架复合物用于修复胎膜破口的研究

Utility of amniotic cells plus collagen Ⅰ complex for fetal membrane healing

目的 探讨羊膜细胞I型胶原蛋白支架复合物用于修复胎膜破口的可行性.方法 将人羊膜间质细胞接种在I型胶原蛋白三维基质中,人羊膜上皮细胞接种在基质的表面,构建羊膜细胞/Ⅰ型胶原蛋白支架复合物以模拟人体羊膜组织结构.DAPI标记三维基质中的细胞核以计数羊膜细胞在三维培养第2,8天的细胞数量.动态测定羊膜细胞/Ⅰ型胶原蛋白复合物体积.培养含有不同数量羊膜间质细胞的羊膜细胞/I型胶原蛋白支架复合物,15 d后检测抗张强度.结果 第2天和第8天的细胞数目分别是（121±5）/cm2和（124±4）/cm2（P〉0.05）.第5天时,I型胶蛋白的面积为原体积的45%,第10天时约为15%,第15天时约为原14%,羊膜间质细胞的存在对于Ⅰ型胶原蛋白基质具有重新塑形的作用（P〈0.01）,羊膜间质细胞可以导致I型胶原蛋白的收缩,抑肽酶和GM6001也不能抑制这种收缩.羊膜细胞/Ⅰ型胶原蛋白复合物抗张强度随人羊膜间质细胞数量的增多而增强（P〈0.01）.结论 羊膜细胞/Ⅰ型胶原蛋白支架复合物有望成为修复胎膜破口的材料.

Objective To study the feasibility of using amniotic cells plus collagen I complex for fetal membrane healing. Methods Human amnion mesenchymal ceils were embedded in three-dimensional （3D） collagen I and epithelial cells placed on the top. The amniotic cells plus collagen I complex was created to mimic the architecture of native amnion. As judged by DAPI staining, the number of amniotic cells cultured in 3D collagen I was counted after incubation for 2 and 8 days. The tensile strength of collagen I with a varying number of amniotic mesenchymal cells was measured after a 15-day incubation. Results At Days 2 and 8, the mean number of mesenchymal cells was （ 121±5 ）/cm2 and （124±4）/cm2 （P 〉0. 05）. The size of collagen I matrix was reduced to 45%, 15%, 14% at Days 5, 10, 15 respectively. Mesenchymal cell was capable of remodeling collagen I （ P 〈 0. 01 ） and it could not be inhibited by GM6001 and/or aprotinin. The tensile strength of amniotic cells plus collagen I scaffold complex was strengthened with an increasing number of amnion mesenchymal cells （ P 〈 0.01 ）. Conclusion The amniotic cells plus collagen I complex may be useful for healing preterm premature rupture of the membrane （PPROM）.