Peng Bou-Yue, Ou Keng-Liang, Liu Chung-Ming, Chu Shu-Fen, Huang Bai-Hung, Cho Yung-Chieh, Saito Takashi, Tsai Chi-Hsun, Hung Kuo-Sheng, Lan Wen-Chien
School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei 110, Taiwan.
Division of Oral and Maxillofacial Surgery, Department of Dentistry, Taipei Medical University Hospital, Taipei 110, Taiwan.
Polymers (Basel). 2022 Aug 21;14(16):3415. doi: 10.3390/polym14163415.
The present study was to investigate the rheological property, printability, and cell viability of alginate−gelatin composed hydrogels as a potential cell-laden bioink for three-dimensional (3D) bioprinting applications. The 2 g of sodium alginate dissolved in 50 mL of phosphate buffered saline solution was mixed with different concentrations (1% (0.5 g), 2% (1 g), 3% (1.5 g), and 4% (2 g)) of gelatin, denoted as GBH-1, GBH-2, GBH-3, and GBH-4, respectively. The properties of the investigated hydrogels were characterized by contact angle goniometer, rheometer, and bioprinter. In addition, the hydrogel with a proper concentration was adopted as a cell-laden bioink to conduct cell viability testing (before and after bioprinting) using Live/Dead assay and immunofluorescence staining with a human corneal fibroblast cell line. The analytical results indicated that the GBH-2 hydrogel exhibited the lowest loss rate of contact angle (28%) and similar rheological performance as compared with other investigated hydrogels and the control group. Printability results also showed that the average wire diameter of the GBH-2 bioink (0.84 ± 0.02 mm (*** p < 0.001)) post-printing was similar to that of the control group (0.79 ± 0.05 mm). Moreover, a cell scaffold could be fabricated from the GBH-2 bioink and retained its shape integrity for 24 h post-printing. For bioprinting evaluation, it demonstrated that the GBH-2 bioink possessed well viability (>70%) of the human corneal fibroblast cell after seven days of printing under an ideal printing parameter combination (0.4 mm of inner diameter needle, 0.8 bar of printing pressure, and 25 °C of printing temperature). Therefore, the present study suggests that the GBH-2 hydrogel could be developed as a potential cell-laden bioink to print a cell scaffold with biocompatibility and structural integrity for soft tissues such as skin, cornea, nerve, and blood vessel regeneration applications.
本研究旨在探究海藻酸钠-明胶复合水凝胶作为一种潜在的载细胞生物墨水用于三维（3D）生物打印应用时的流变学性质、可打印性和细胞活力。将2 g海藻酸钠溶解于50 mL磷酸盐缓冲盐溶液中，与不同浓度（1%（0.5 g）、2%（1 g）、3%（1.5 g）和4%（2 g））的明胶混合，分别记为GBH-1、GBH-2、GBH-3和GBH-4。通过接触角测量仪、流变仪和生物打印机对所研究水凝胶的性质进行表征。此外，采用浓度合适的水凝胶作为载细胞生物墨水，使用活/死检测法以及用人角膜成纤维细胞系进行免疫荧光染色，对（生物打印前后的）细胞活力进行测试。分析结果表明，与其他所研究的水凝胶和对照组相比，GBH-2水凝胶的接触角损失率最低（28%），且流变学性能相似。可打印性结果还表明，GBH-2生物墨水打印后的平均线径（0.84±0.02 mm（***p<0.001））与对照组（0.79±0.05 mm）相似。此外，可由GBH-2生物墨水制备细胞支架，且打印后24小时内其形状完整性得以保持。对于生物打印评估，结果表明在理想的打印参数组合（内径0.4 mm的针头、0.8 bar的打印压力和25°C的打印温度）下，打印七天后GBH-2生物墨水对人角膜成纤维细胞具有良好的活力（>70%）。因此，本研究表明GBH-2水凝胶可被开发为一种潜在的载细胞生物墨水，用于打印具有生物相容性和结构完整性的细胞支架，以应用于皮肤、角膜、神经和血管等软组织的再生。
