以壳聚糖复合膜为载体支架体外培养家兔角膜内皮细胞的实验研究

Experimental Studies on Chitosan Blend Membrane as Scaffold Carriers for Cultivating Rabbit Corneal Endothelial Cells In Vitro

利用4ha(壳聚糖-透明质酸)、631ha(壳聚糖-透明质酸)和631s(壳聚糖-硫酸软骨素)复合膜为载体支架培养无转染家兔角膜内皮细胞(RCEC)系细胞,根据细胞贴附、形态变化、生长以及形成单层的情况研究了三种载体支架与RCEC的生物相容性。结果表明,培养在4ha和631ha膜上的RCEC,出现了细胞局部聚集和脱膜现象,尤其是培养在631ha膜的RCEC还出现了成片脱膜现象;此外,RCEC在4ha膜上无法形成细胞单层,但在631ha膜上48h后可基本形成细胞单层;而培养在631s膜上的RCEC,细胞形态呈梭形、分布均匀、生长状态好,48h后已基本形成细胞单层,72h后形成了完整的细胞单层,且细胞与膜结合紧密。可见,631s膜很可能是RCEC体外培养的理想载体,有望用于组织工程角膜内皮研制的载体支架材料。

Un-transfected rabbit corneal endothelial cells （RCECs） were cultivated,using chitosan blend membrane 4ha （chitosan-hyaluronie acid）, 631ha （chitosan-hyaluronic acid） and 631s （chitosan-chondroitine sulfate） as scaffold carriers. Their biocompatibilities were studied in regard to ceil adherence, morphological changes, growth status and monolayer forming abilities. The results indicated that RCECs cultivated on 4ha and 631ha carriers tended to be aggregated and even desquamated to some extent in local areas, and even more severely on 631ha carrier. And the RCECs cultivated on 631ha carrier could form almost a monolayer 48h later, and those on 4ha carrier could not. Contrarily, the RCECs cultivated on 631s carrier were evenly distributed and were in good status of growth with a good adherence and fibroblast-like morphology which could form almost a monolayer 48h later. And a complete monolayer was formed and was tightly attached to the 631s carrier 72h later. From the above results, it can be concluded that 631s carrier is most probably an ideal scaffold carrier for RCEC cultivation. 631s carrier may have the poteintial for use in the development of tissue-engineered rabbit corneal endothelium.