Kumari Shikha, Singh Bhisham Narayan, Srivastava Pradeep
School of Biochemical Engineering, Indian Institute of Technology, Varanasi, 221005 India.
3 Biotech. 2019 Mar;9(3):102. doi: 10.1007/s13205-019-1624-9. Epub 2019 Feb 21.
Development of new and effective scaffold continues to be an area of intense research in skin tissue engineering. The objective of this study was to study the effect of copper nanoparticles over physico-chemical properties of the chitosan and gelatin composite scaffolds for skin tissue engineering. The copper-doped scaffolds were prepared using freeze-drying method. Chitosan and gelatin were taken in varied composition with 0.01%, 0.02%, and 0.03% Cu nanoparticles. The physico-chemical properties of the copper nanoparticles and the scaffolds were analyzed using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy. Porosity of the scaffolds was measured by liquid displacement method and hemocompatibility was tested using goat blood. SEM micrographs of the scaffolds displayed the interconnected pores which ranged between 25 and 40 µm. This average pore size was later enhanced to 95 µm after the addition of copper nanoparticles. Cell viability assay was performed to ensure the growth and proliferation of the skin cells over the scaffolds. FTIR, EDS, and XRD analysis of scaffolds confirmed the presence of copper in the chitosan-based scaffolds. Porosity measurement showed the interconnectivity between pores which ranged between 65 and 88% as required for skin tissue engineering application. The degradation study of the scaffolds was done which depicted that, after the addition of copper nanoparticles with 0.03%, degradation rate was decreased. SEM and cytocompatibility assay on all scaffolds showed the cell adhesion and proliferation on the scaffolds which was not affected after addition of copper nanoparticles. Oxidative stress evaluation was done to study the effect of copper nanoparticles on the cells which showed that there was no such production of ROS in the scaffolds. Hence, scaffolds prepared after doping of copper nanoparticles show suitable physico-chemical and biological properties for skin tissue engineering application.
新型有效支架的研发仍是皮肤组织工程领域的一个热门研究方向。本研究的目的是研究铜纳米颗粒对用于皮肤组织工程的壳聚糖和明胶复合支架物理化学性质的影响。采用冷冻干燥法制备了掺铜支架。壳聚糖和明胶以不同的组成比例与0.01%、0.02%和0.03%的铜纳米颗粒混合。使用傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）、扫描电子显微镜（SEM）和透射电子显微镜对铜纳米颗粒和支架的物理化学性质进行了分析。通过液体置换法测量支架的孔隙率，并使用山羊血测试血液相容性。支架的SEM显微照片显示出相互连通的孔隙，孔径在25至40微米之间。添加铜纳米颗粒后，该平均孔径随后增大至95微米。进行细胞活力测定以确保皮肤细胞在支架上的生长和增殖。支架的FTIR、能谱分析（EDS）和XRD分析证实了壳聚糖基支架中存在铜。孔隙率测量显示孔隙之间的连通性在皮肤组织工程应用所需的65%至88%之间。对支架进行了降解研究，结果表明，添加0.03%的铜纳米颗粒后，降解速率降低。所有支架的SEM和细胞相容性测定表明，支架上有细胞粘附和增殖，添加铜纳米颗粒后未受影响。进行了氧化应激评估以研究铜纳米颗粒对细胞的影响，结果表明支架中没有产生活性氧。因此，掺杂铜纳米颗粒后制备的支架显示出适用于皮肤组织工程应用的物理化学和生物学性质。
