动-静态循序法构建自体组织工程化结膜重建眼表

Conjunctival reconstruction provides healthy conjunctival microenvironment. It is an essential part of ocular surface reconstruction, especially in patients with total surface involvement. However, lack of suitable donor tissue and disappointing clinical effect are the major unsolved problems. In this study we plan to construct tissue-engineered conjunctiva for clinical application. Phospholipase A2 and sodium deoxycholate are used to prepare acellular porcine conjunctival stroma as a tissue engineering scaffold. Autologous conjunctiva explants are cultured to amplify conjunctiva epithelium. Then conjunctiva epithelium is implanted on the prepared scaffold to construct tissue-engineered conjunctiva in the dynamic and static physiological culturing system with appropriate parameters. The histological structures and physiological functions of the tissue-engineered conjunctiva are assessed. Then tissue-engineered conjunctiva is transplanted to a conjunctival defect animal model. Amniotic membrane contact by sutureless fixing device (Authorized Patent: ZL 200610156533.4) is combined with tissue-engineered conjunctiva for conjunctival reconstruction. The associated parameters are ajusted according to clinical effect of conjunctival reconstruction.

结膜重建可提供良好的结膜微环境，是眼表重建的基础和必须。发展组织工程化结膜可协助解决结膜重建中供体材料不足和疗效不佳等难题。本项目使用磷脂酶A2联合脱氧胆酸钠脱细胞的方法，制备满足生物学和物理力学要求的异种脱细胞结膜基质载体；以扩增的自体结膜上皮细胞为种子细胞；采用动-静态联合循序培养的方法，通过调整培养系统中生长因子的比例、浓度和培养时间等系列参数，促进自体结膜上皮细胞在脱细胞结膜基质支架上增殖、粘附和分化，从而构建出可移植的组织工程化结膜。将所构建的组织工程化结膜联合前期发明的眼表生物膜固定装置制成的无缝线羊膜接触镜(授权专利号ZL 200610156533.4)，用于结膜缺损动物模型，完成结膜重建。根据检测结果和结膜重建的效果进一步调整和确定各项参数，达到最佳的眼表结膜重建效果。

结膜重建可提供良好的结膜微环境，是眼表重建的基础和必须。发展组织工程化结膜可协助解决结膜重建中供体材料不足和疗效不佳等难题。本项目使用磷脂酶A2联合脱氧胆酸钠脱细胞的方法，制备满足生物学和物理力学要求的异种脱细胞结膜基质载体；以扩增的自体结膜上皮细胞为种子细胞；采用动-静态联合循序培养的方法，通过调整构建参数，促进自体结膜上皮细胞在脱细胞结膜基质支架上增殖、粘附和分化，从而构建出可移植的组织工程化结膜。将所构建的组织工程化结膜联合前期的眼表生物膜固定装置制成的无缝线羊膜接触镜(授权专利号ZL 200610156533.4)，用于结膜缺损动物模型，完成结膜重建。并根据结膜重建的效果进一步调整和确定各项参数，达到更好的眼表结膜重建效果。