胶原-硫酸软骨素-透明质酸真皮支架材料修复兔角膜基质缺损的研究

Fabrication of Collagen-Chondroitin Sulfate-Hyaluronic Acid Scaffold and Its Biological Characteristics for Cornea Stroma Tissue Engineering

目的探讨以角膜基质细胞与胶原-硫酸软骨素-透明质酸(Collagen-chondroitin sulfate-hyaluronic acid,Co-CS-HA)真皮支架材料复合共培养构建组织工程化角膜基质的可行性,为寻找理想的组织工程角膜支架提供依据。方法先行制备Co-CS-HA真皮支架材料,对其进行扫描电镜、孔隙率、体外降解性、力学性能和细胞增殖的检测和分析。培养兔角膜基质细胞,将其与上述材料复合共培养构建组织工程角膜基质,行兔角膜板层移植。术后大体观察植片和新生血管状况,并于术后第8周取材行组织学观察。结果所制备的Co-CS-HA真皮支架材料为白色海绵状结构,其孔径为(109±11)μm,孔隙均匀并有广泛的交通孔,孔隙率为(94.75±0.39)%。体外降解性和力学性能检测结果显示材料的强度和抵抗酶解的能力显著提高。MTS结果表明,材料对角膜基质细胞的增殖有明显促进作用。兔角膜板层移植后,组织工程角膜植片与宿主角膜组织相融性良好,能够部分修复并填充角膜基质缺损,但直到8周取材时仍留有大量的角膜新生血管。结论本实验室制备的Co-CS-HA真皮支架材料符合支架材料的理化和生物学要求,可一定程度修复兔角膜基质缺损,但在维持角膜的透光度方面仍存有缺陷,眼科应用还需进行更深入研究。

Objective To develop a scaffold material containing collagen-chondroitin sulfate-hyaluronic acid （Co-CS- HA） and corneal stromal cells corneal stroma tissue engineering. Methods Fabricated scaffold of Co-CS-HA for dermal tissue engineering and to evaluated its characteristics in vitro. The tissue- engineered corneal stroma was constructed by co-culturing rabbit keratocytes on Co-CS-HA scaffold. It was then transplanted into corneal stroma of rabbit. This tissue-engineered corneal stroma was observed and harvested after 8 weeks for gross and histological evaluation. Results The scaffold of CoCS-HA was white sponginess structure and had uniform, and widely inter-connected pores with mean diameters of （109±11） μm and adequate porosity of about 94%. Mechanical properties and biodegradation rates assessment indicated that it was also more degradation-resistant and had higher elastic modulus than collagen sponge. The porous scaffold has a good biocompatibility for the attachment and proliferation of the cornea stroma cells without cytotoxicity. After implanted into corneal stroma of rabbit, the corneal stroma equivalent could partially repair defect. But there had been a plenty of corneal new vessels remained until 8 weeks. Conclusion The scaffold of Co-CS-HA for dermal tissue engineering can repair corneal stroma defect to some extent, but there were still many new vessels left. So it should be studied deeply in clinical application of ophthalmology.