Bhowmick Sirsendu, Scharnweber Dieter, Koul Veena
Max Bergmann Center of Biomaterials, Technische Universität Dresden, Budapester Straße 27, 01069 Dresden, Germany; Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Hauz Khas, 110016 New Delhi, India.
Max Bergmann Center of Biomaterials, Technische Universität Dresden, Budapester Straße 27, 01069 Dresden, Germany.
Biomaterials. 2016 May;88:83-96. doi: 10.1016/j.biomaterials.2016.02.034. Epub 2016 Feb 24.
Fortifying the scaffold with bioactive molecules and glycosaminoglycans (GAGs), is an efficient way to design new generation tissue engineered biomaterials. In this study, we evaluated the synergistic effect of electrospun nanofibrous composite scaffold (cationic gelatin/hyaluronan/chondroitin sulfate) loaded with sericin and, contact co-culture of human mesenchymal stem cells (hMSCs)-keratinocytes on hMSCs' differentiation towards epithelial lineage. Cationic gelatin is prepared with one step novel synthesis process by grafting quaternary ammonium salts to the backbone of gelatin. Release kinetics studies showed that Fickian diffusion is the major release mechanism for both GAGs and sericin/gelatin. In vitro biocompatibility of the electrospun scaffold was evaluated in terms of LDH and DNA quantification assay on human foreskin fibroblast, human keratinocyte and hMSC. Significant proliferation (∼ 4-6 fold) was detected after culturing all three cell on the electrospun scaffold containing sericin. After 5 days of contact co-culture, results revealed that electrospun scaffold containing sericin promote epithelial differentiation of hMSC in terms of several protein markers (keratin 14, ΔNp63α and Pan-cytokeratin) and gene expression of some dermal proteins (keratin 14, ΔNp63α). Findings of this study will foster the progress of current skin tissue engineering scaffolds by understanding the skin regeneration and wound healing process.
用生物活性分子和糖胺聚糖（GAGs）强化支架，是设计新一代组织工程生物材料的有效方法。在本研究中，我们评估了负载丝胶蛋白的电纺纳米纤维复合支架（阳离子明胶/透明质酸/硫酸软骨素）以及人间充质干细胞（hMSCs）与角质形成细胞的接触共培养对hMSCs向上皮谱系分化的协同作用。阳离子明胶通过将季铵盐接枝到明胶主链上，采用一步法新颖合成工艺制备。释放动力学研究表明，菲克扩散是GAGs和丝胶蛋白/明胶的主要释放机制。通过对人包皮成纤维细胞、人角质形成细胞和hMSC进行乳酸脱氢酶（LDH）和DNA定量测定，评估了电纺支架的体外生物相容性。在含有丝胶蛋白的电纺支架上培养所有三种细胞后，检测到显著的增殖（约4 - 6倍）。接触共培养5天后，结果显示，含有丝胶蛋白的电纺支架在几种蛋白质标志物（角蛋白14、ΔNp63α和泛细胞角蛋白）以及一些真皮蛋白（角蛋白14、ΔNp63α）的基因表达方面促进了hMSC的上皮分化。本研究结果将通过了解皮肤再生和伤口愈合过程，推动当前皮肤组织工程支架的发展。
