School of Pharmacy, University of Reading, Reading, RG6 6UB, United Kingdom.
J Biomed Mater Res A. 2010 Nov;95(2):447-53. doi: 10.1002/jbm.a.32861.
We compare the use of plastically compressed collagen gels to conventional collagen gels as scaffolds onto which corneal limbal epithelial cells (LECs) are seeded to construct an artificial corneal epithelium. LECs were isolated from bovine corneas (limbus) and seeded onto either conventional uncompressed or novel compressed collagen gels and grown in culture. Scanning electron microscopy (SEM) results showed that fibers within the uncompressed gel were loose and irregularly ordered, whereas the fibers within the compressed gel were densely packed and more evenly arranged. Quantitative analysis of LECs expansion across the surface of the two gels showed similar growth rates (p > 0.05). Under SEM, the LECs, expanded on uncompressed gels, showed a rough and heterogeneous morphology, whereas on the compressed gel, the cells displayed a smooth and homogeneous morphology. Transmission electron microscopy (TEM) results showed the compressed scaffold to contain collagen fibers of regular diameter and similar orientation resembling collagen fibers within the normal cornea. TEM and light microscopy also showed that cell-cell and cell-matrix attachment, stratification, and cell density were superior in LECs expanded upon compressed collagen gels. This study demonstrated that the compressed collagen gel was an excellent biomaterial scaffold highly suited to the construction of an artificial corneal epithelium and a significant improvement upon conventional collagen gels.
我们比较了使用塑性压缩胶原凝胶和传统胶原凝胶作为支架,将角膜缘上皮细胞(LEC)接种到支架上,以构建人工角膜上皮。LEC 从牛角膜(角膜缘)中分离出来,接种到传统的未压缩或新型压缩胶原凝胶上,并在培养中生长。扫描电子显微镜(SEM)结果表明,未压缩凝胶内的纤维松散且排列不规则,而压缩凝胶内的纤维则紧密排列且排列更均匀。对两种凝胶表面上 LEC 扩展的定量分析表明,生长速率相似(p>0.05)。在 SEM 下,在未压缩凝胶上扩展的 LEC 显示出粗糙且不均匀的形态,而在压缩凝胶上,细胞显示出光滑且均匀的形态。透射电子显微镜(TEM)结果表明,压缩支架包含具有相似取向的规则直径的胶原纤维,类似于正常角膜中的胶原纤维。TEM 和光学显微镜还表明,在压缩胶原凝胶上扩展的 LEC 中,细胞-细胞和细胞-基质附着、分层和细胞密度更好。这项研究表明,压缩胶原凝胶是一种非常适合构建人工角膜上皮的优秀生物材料支架,比传统胶原凝胶有显著的改进。