Middelkoop E, de Vries H J, Ruuls L, Everts V, Wildevuur C H, Westerhof W
Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, The Netherlands.
Cell Tissue Res. 1995 May;280(2):447-53. doi: 10.1007/BF00307818.
We describe an in vitro model that we have used to evaluate dermal substitutes and to obtain data on cell proliferation, the rate of degradation of the dermal equivalent, contractibility and de novo synthesis of collagen. We tested three classes of collagenous materials: (1) reconstituted non-crosslinked collagen, (2) reconstituted collagen that was chemically crosslinked with either glutaraldehyde, aluminium alginate or acetate, and (3) native collagen fibres, with or without other extracellular matrix molecules (elastin hydrolysate, hyaluronic acid or fibronectin). The non-crosslinked reconstituted collagen was degraded rapidly by human fibroblasts. The chemically crosslinked materials proved to be cytotoxic. Native collagen fibres were stable. In the absence of ascorbic acid, the addition of elastin hydrolysate to this type of matrix reduced the rate of collagen degradation. Both elastin hydrolysate and fibronectin partially prevented fibroblast-mediated contraction. Hyaluronic acid was only slightly effective in reducing the collagen degradation rate and more fibroblast-mediated contraction of the material was found than for the native collagen fibres with elastin hydrolysate and fibronectin. In the presence of ascorbate, collagen synthesis was enhanced in the native collagen matrix without additions and in the material containing elastin hydrolysate, but not in the material with hyaluronic acid. These results are indicative of the suitability of tissue substitutes for in vivo application.
我们描述了一种体外模型，该模型用于评估真皮替代物，并获取有关细胞增殖、真皮替代物降解速率、收缩性以及胶原蛋白从头合成的数据。我们测试了三类胶原质材料：（1）重组非交联胶原蛋白，（2）用戊二醛、海藻酸铝或醋酸化学交联的重组胶原蛋白，以及（3）天然胶原纤维，有无其他细胞外基质分子（弹性蛋白水解物、透明质酸或纤连蛋白）。非交联重组胶原蛋白被人成纤维细胞迅速降解。化学交联材料被证明具有细胞毒性。天然胶原纤维是稳定的。在缺乏抗坏血酸的情况下，向此类基质中添加弹性蛋白水解物可降低胶原蛋白降解速率。弹性蛋白水解物和纤连蛋白均可部分阻止成纤维细胞介导的收缩。透明质酸在降低胶原蛋白降解速率方面效果甚微，且发现其比添加了弹性蛋白水解物和纤连蛋白的天然胶原纤维有更多的成纤维细胞介导的收缩。在有抗坏血酸存在的情况下，未添加其他成分的天然胶原基质以及含有弹性蛋白水解物的材料中胶原蛋白合成增强，但含有透明质酸的材料中则未增强。这些结果表明组织替代物适用于体内应用。
