猪角膜脱细胞基质的制备及生物相容性的实验研究

Preparation of porcine cornea acellular matrix and its biocompatibility

目的探讨猪角膜脱细胞基质(CACM)的制备方法,评价其组织学特性和生物相容性。方法应用1%TritonX-100去垢剂振洗,经冷冻干燥处理得到猪CACM,通过苏木精-伊红染色、扫描电镜观察行组织学检测。将猪CACM植入兔角膜板层间观察10周,取材做组织切片,评价其生物组织相容性。结果组织学检测证实角膜细胞完全脱净,胶原纤维排列疏松,板层结构同正常角膜;扫描电镜CACM表面未见细胞结构,纵切面上胶原板层间出现清晰的空穴状裂隙;CACM植入兔角膜层间后,观察期内未见明显排异反应。结论TritonX-100可以有效地脱净角膜细胞,保存胶原排列的结构特征,经过冷冻干燥可形成多孔隙且胶原排列疏松的板层结构,适合作为载体材料,猪CACM与兔角膜生物相容性好。

Objective As a tissue engineering material, acellular matrix is extensively used to the repairing and reconstruction of tissue,and the research and utilization of corneal acellular matrix is seldom. This paper was to investigate the method and process of obtaining porcine Cornea Acellular Matrix （CACM） with fine reticular collagen and evaluate its biocompatibillty and safety, Methods Porcine corneas were treated using detergent 1% TritonX-100 and freeze-drying process to prepare CACM. HE staining and scanning electron microscope were performed for the histologic examination of CACM. A part of porcine CACM lamina was implanted into the rabbit＇ s corneal lamina. Rabbits were sacrificed and cross-section of corneas were prepared for HE staining at the end of 6-week. Results The histologic examination indicated that porcine cornea cells were clearly extracted, and fiber arranged in a looser fashion. Lamina structure resembled to that of the normal. No cells residua was seen,and a porous character was exhibited under the scan electron microscope. No rejection was observed after implanting into the lamina of CACM, showing a good biocompatibility of CACM to cornea of rabbit. Conclusion 1% TritonX-100 alone can effectively remove cells of cornea stroma and ensure the structure features of corneal collagen. Freeze-drying makes biomaterial porous and collagen loose. CACM has equal biocompatibility to scaffold.