1 Tissue Engineering Centre, Faculty of Medicine, UKM Medical Centre , Cheras, Malaysia .
2 Industrial Technology Division, Agensi Nuklear Malaysia , Kajang, Malaysia .
Tissue Eng Part C Methods. 2018 Jun;24(6):368-378. doi: 10.1089/ten.TEC.2017.0447. Epub 2018 May 24.
Fabrication of composite scaffolds is one of the strategies proposed to enhance the functionality of tissue-engineered scaffolds for improved tissue regeneration. By combining multiple elements together, unique biomimetic scaffolds with desirable physical and mechanical properties can be tailored for tissue-specific applications. Despite having a highly porous structure, the utility of electrospun fibers (EF) as scaffold is usually hampered by their insufficient mechanical strength. In this study, we attempted to produce a mechanically competent scaffold with cell-guiding ability by fabricating aligned poly lactic-co-glycolic acid (PLGA) fibers on decellularized human amniotic membrane (HAM), known to possess favorable tensile and wound healing properties. Decellularization of HAM in 18.75 μg/mL of thermolysin followed by a brief treatment in 0.25 M sodium hydroxide efficiently removed the amniotic epithelium and preserved the ultrastructure of the underlying extracellular matrix. The electrospinning of 20% (w/v) PLGA 50:50 polymer on HAM yielded beadless fibers with straight morphology. Subsequent physical characterization revealed that EF-HAM scaffold with a 3-min fabrication had the most aligned fibers with the lowest fiber diameter in comparison with EF-HAM 5- and 7-min scaffolds. Hydrated EF-HAM scaffolds with 3-min deposition had a greater tensile strength than the other scaffolds despite having thinner fibers. Nevertheless, wet HAM and EF-HAMs regardless of the fiber thicknesses had a significantly lower Young's modulus, and hence, a higher elasticity compared with dry HAM and EF-HAMs. Biocompatibility analysis showed that the viability and migration rate of skeletal muscle cells on EF-HAMs were similar to control and HAM alone. Skeletal muscle cells seeded on HAM were shown to display random orientation, whereas cells on EF-HAM scaffolds were oriented along the alignment of the electrospun PLGA fibers. In summary, besides having good mechanical strength and elasticity, EF-HAM scaffold design decorated with aligned fiber topography holds a promising potential for use in the development of aligned tissue constructs.
制备复合材料支架是增强组织工程支架功能以促进组织再生的策略之一。通过将多种元素结合在一起，可以为特定组织应用定制具有理想物理和机械性能的独特仿生支架。尽管具有高度多孔的结构，但电纺纤维（EF）作为支架的用途通常受到其机械强度不足的限制。在这项研究中，我们试图通过在具有良好拉伸和伤口愈合性能的脱细胞人羊膜（HAM）上制造对齐的聚乳酸-共-羟基乙酸（PLGA）纤维来制造具有细胞导向能力的机械性能良好的支架。在 18.75μg/mL 耐热溶菌酶中对 HAM 进行脱细胞处理，然后在 0.25M 氢氧化钠中进行短暂处理，有效地去除了羊膜上皮细胞并保留了下伏细胞外基质的超微结构。在 HAM 上电纺 20%（w/v）PLGA 50:50 聚合物可得到具有直形态的无珠纤维。随后的物理特性分析表明，与 EF-HAM 5-和 7-分钟支架相比，具有 3 分钟制备时间的 EF-HAM 支架具有最对齐的纤维和最低的纤维直径。尽管具有更细的纤维，但具有 3 分钟沉积的水合 EF-HAM 支架具有比其他支架更高的拉伸强度。然而，无论纤维厚度如何，湿 HAM 和 EF-HAMs 的杨氏模量均显著降低，因此与干 HAM 和 EF-HAMs 相比具有更高的弹性。生物相容性分析表明，骨骼肌细胞在 EF-HAMs 上的存活率和迁移率与对照和单独的 HAM 相似。在 HAM 上接种的骨骼肌细胞表现出随机取向，而在 EF-HAM 支架上接种的细胞则沿着电纺 PLGA 纤维的对齐方向取向。总之，除了具有良好的机械强度和弹性外，具有对齐纤维形貌的 EF-HAM 支架设计在开发对齐组织构建体方面具有很大的潜力。
