Abstract

In tissue engineering bioresorbable polymers are commonly used to mimetize a damaged tissue characteristics. In spite of that, we did not find a comparative study of the response of the same cell to dense, porous, and fibrous biomaterials. The present work aimed to study the in vitro of pure polymers [poly (ε-caprolactone) (PCL) and poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV)] and their 75:25 blend in dense, porous, and fibrous forms in Vero cells, a fibroblast cell line. The biomaterials were characterized morphologically by scanning electron microscopy (SEM). The MTT assay was used for analysis of possible cytotoxicity. Vero cells were cultured on biomaterials for 120 h and submitted to SEM, morphological and cytochemical analyses. The dyes used were crystal violet (morphology), toluidine blue at pH 4.0 (for nucleic acids and glycosaminoglycans) and 2.5 (for glycosaminoglycans only), periodic acid-Schiff reactive (for neutral sugars), xylidine ponceau at pH 2.5 (for total proteins), and picrosirius/hematoxylin (for selective collagen analysis). The biomaterials studied were not considered citocytotoxic. The cells were able to adhere to and proliferate on the substrates. The cytochemical data indicate that the cells have high functional activity, with high protein synthesis on the scaffolds but do not accumulate acid or neutral sugars. The cells on the fibrous polymers appear to be more elongated, following the shape of the fibers. Regarding the scaffold form (porous or fibrous), the in vitro data do not show great superiority of one type to the other. Then, all the polymers tested are suitable for tissue engineering.

摘要

在组织工程中，生物可吸收聚合物通常用于模拟受损组织的特性。尽管如此，我们没有找到关于同一细胞对致密、多孔和纤维状生物材料的反应的比较研究。目前的工作旨在研究体外纯聚合物 [聚 (ε-己内酯) (PCL) 和聚 (羟基丁酸-羟基戊酸共聚物) (PHBV)] 及其 75:25 在 Vero 细胞（一种成纤维细胞系）中以致密、多孔和纤维形式混合。通过扫描电子显微镜 (SEM) 对生物材料进行形态学表征。MTT 测定用于分析可能的细胞毒性。Vero 细胞在生物材料上培养 120 小时，并进行 SEM、形态学和细胞化学分析。使用的染料为结晶紫（形态学）、pH 4.0 的甲苯胺蓝（用于核酸和糖胺聚糖）和 2.5（仅用于糖胺聚糖）、高碘酸席夫反应性染料（用于中性糖）、二甲苯胺蓝 pH 2.5（用于总蛋白质） ), and picrosirius/hematoxylin (用于选择性胶原蛋白分析)。所研究的生物材料不被认为具有细胞毒性。细胞能够粘附在基质上并在基质上增殖。细胞化学数据表明细胞具有高功能活性，在支架上具有高蛋白质合成但不积累酸性或中性糖。根据纤维的形状，纤维聚合物上的细胞似乎更细长。关于支架形式（多孔或纤维），体外数据并未显示一种类型优于另一种类型。然后，所有测试的聚合物都适用于组织工程。