3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, University of Minho, Avepark - Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal.
Biomater Sci. 2019 Nov 19;7(12):5451-5466. doi: 10.1039/c9bm01098j.
Although surgical management of peripheral nerve injuries (PNIs) has improved over time, autografts are still the current "gold standard" treatment for PNIs, which presents numerous limitations. In an attempt to improve natural biomaterial-based nerve guidance conduits (NGCs), chitosan (CHT), a derivative of the naturally occurring biopolymer chitin, has been explored for peripheral nerve regeneration (PNR). In addition to CHT, keratin has gained enormous attention as a biomaterial and tissue engineering scaffolding. In this study, biomimetic CHT/keratin membranes were produced using a solvent casting technique. These membranes were broadly characterized in terms of their surface topography and physicochemical properties, with techniques such as Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), contact angle, weight loss and water uptake measurements, Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). Biological in vitro assays were also performed, where a preliminary cytotoxicity screening with the L929 fibroblast cell line revealed that the membranes and respective materials are suitable for cell culture. In addition, Schwann cells, fibroblasts and endothelial cells were directly seeded in the membranes. Quantitative and qualitative assays revealed that the addition of keratin enhanced cell viablity and adhesion. Based on the encouraging in vitro results, the in vivo angiogenic/antiangiogenic potential of CHT and CHT/keratin membranes was assessed, using an optimized chick embryo chorioallantoic membrane assay, where higher angiogenic responses were seen in keratin-enriched materials. Overall, the obtained results indicate the higher potential of CHT/keratin membranes for guided tissue regeneration applications in the field of PNR.
虽然外周神经损伤 (PNI) 的手术治疗随着时间的推移有所改善，但自体移植物仍然是 PNI 的当前“金标准”治疗方法，存在许多局限性。为了改善基于天然生物材料的神经引导导管 (NGC)，壳聚糖 (CHT)，一种天然存在的生物聚合物几丁质的衍生物，已被探索用于周围神经再生 (PNR)。除了 CHT 之外，角蛋白作为生物材料和组织工程支架也受到了极大的关注。在这项研究中，使用溶剂铸造技术制备了仿生 CHT/角蛋白膜。使用傅里叶变换红外光谱 (FTIR)、差示扫描量热法 (DSC)、接触角、失重和吸水率测量、扫描电子显微镜 (SEM) 和原子力显微镜 (AFM) 等技术对这些膜进行了广泛的表面形貌和物理化学性质的表征。还进行了生物体外检测，用 L929 成纤维细胞系进行初步细胞毒性筛选表明，这些膜和相应的材料适合细胞培养。此外，还将雪旺细胞、成纤维细胞和内皮细胞直接接种在膜中。定量和定性检测表明，角蛋白的添加增强了细胞活力和粘附性。基于令人鼓舞的体外结果，使用优化的鸡胚绒毛尿囊膜试验评估了 CHT 和 CHT/角蛋白膜的血管生成/抗血管生成潜力，在富含角蛋白的材料中观察到更高的血管生成反应。总的来说，获得的结果表明 CHT/角蛋白膜在 PNR 领域的组织引导再生应用中具有更高的潜力。
