Sadeghi A R, Nokhasteh S, Molavi A M, Khorsand-Ghayeni M, Naderi-Meshkin H, Mahdizadeh A
Materials Research Group, Iranian Academic Center for Education, Culture and Research, (ACECR), Mashhad Branch, Mashhad, Iran.
Materials Research Group, Iranian Academic Center for Education, Culture and Research, (ACECR), Mashhad Branch, Mashhad, Iran.
Mater Sci Eng C Mater Biol Appl. 2016 Sep 1;66:130-137. doi: 10.1016/j.msec.2016.04.073. Epub 2016 Apr 23.
In skin tissue engineering, surface feature of the scaffolds plays an important role in cell adhesion and proliferation. In this study, non-woven fibrous substrate based on poly (lactic-co-glycolic acid) (PLGA) (75/25) were hydrolyzed in various concentrations of NaOH (0.05N, 0.1N, 0.3N) to increase carboxyl and hydroxyl groups on the fiber surfaces. These functional groups were activated by EDC/NHS to create chemical bonding with collagen. To improve bioactivity, the activated substrates were coated with a collagen solution (2mg/ml) and cross-linking was carried out using the EDC/NHS in MES buffer. The effectiveness of the method was evaluated by contact angle measurements, porosimetry, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), tensile and degradation tests as well as in vitro cell attachment and cytotoxicity assays. Cell culture results of human dermal fibroblasts (HDF) and keratinocytes cell line (HaCat) revealed that the cells could attach to the scaffold. Further investigation with MTT assay showed that the cell proliferation of HaCat significantly increases with collagen coating. It seems that sufficient stability of collagen on the surface due to proper chemical bonding and cross-linking has increased the bioactivity of surface remarkably which can be promising for bioengineered skin applications.
在皮肤组织工程中,支架的表面特征在细胞黏附和增殖方面起着重要作用。在本研究中,基于聚(乳酸 - 乙醇酸)共聚物(PLGA)(75/25)的非织造纤维基质在不同浓度的NaOH(0.05N、0.1N、0.3N)中水解,以增加纤维表面的羧基和羟基。这些官能团通过EDC/NHS活化,以与胶原蛋白形成化学键。为提高生物活性,将活化后的基质用胶原蛋白溶液(2mg/ml)包被,并在MES缓冲液中使用EDC/NHS进行交联。通过接触角测量、孔隙率测定、扫描电子显微镜(SEM)、傅里叶变换红外光谱(FTIR)、拉伸和降解测试以及体外细胞附着和细胞毒性测定来评估该方法的有效性。人真皮成纤维细胞(HDF)和角质形成细胞系(HaCat)的细胞培养结果表明,细胞能够附着在支架上。进一步用MTT法研究表明,胶原蛋白包被后HaCat细胞的增殖显著增加。由于适当的化学键合和交联,胶原蛋白在表面具有足够的稳定性,这似乎显著提高了表面的生物活性,这对于生物工程皮肤应用可能是有前景的。