Department of Bionanotechnology and Bio-Convergence Engineering, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju-si, Jeollabuk-do 54896, Republic of Korea.
Department of Polymer-Nano Science and Technology, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju-si, Jeollabuk-do 54896, Republic of Korea.
ACS Appl Bio Mater. 2021 Feb 15;4(2):1771-1782. doi: 10.1021/acsabm.0c01516. Epub 2021 Feb 1.
In this study, dopamine-functionalized gellan gum (DFG) hydrogel was prepared as a carrier for retinal pigment epithelium (RPE) cell delivery a carbodiimide reaction. The carboxylic acid of gellan gum (GG) was replaced with catechol in a 21.3% yield, which was confirmed by NMR. Sol fraction and weight loss measurements revealed that dopamine improved degradability in the GG hydrogel. Measurements of the viscosity, injection force, and compressibility also showed that dopamine-functionalized GG hydrogels had more desirable rheological/mechanical properties for improving injectability. These characteristics were confirmed to arise from the GG's helix structure loosened by the dopamine's bulky nature. Moreover, dopamine's hydrophilic characteristics were confirmed to create a more favorable microenvironment for cell growth by promoting swelling capability and cell attachment. This improved biocompatibility became more pronounced when the hydrophilicity of dopamine was combined with a larger specific surface area stemming from the less porous structure of the dopamine-grafted hydrogels. This effect was apparent from the live/dead staining images of the as-prepared hydrogels. Meanwhile, the nonionic cross-linked DFG (DG) hydrogel showed the lowest protein expression in the immunofluorescence staining images obtained after 28 days of culture, supporting that it had the highest degradability and associated cell-releasing ability. That tendency was also observed in the gene expression data acquired by real-time polymerase chain reaction (RT-PCR) analysis. RT-PCR analysis also revealed that the DG hydrogel carrier could upregulate the visual function-related gene of RPE. Overall, the DG hydrogel system demonstrated its feasibility as a carrier of RPE cells and its potential as a means of improving visual function.
在这项研究中,多巴胺功能化的结冷胶(DFG)水凝胶被制备为视网膜色素上皮(RPE)细胞递送的载体 通过碳化二亚胺反应。结冷胶(GG)的羧酸以 21.3%的产率被邻苯二酚取代,这通过 NMR 得到了证实。溶胶分数和重量损失测量表明,多巴胺提高了 GG 水凝胶的降解性。粘度、注射力和可压缩性的测量也表明,多巴胺功能化的 GG 水凝胶具有更理想的流变/力学性能,可提高可注射性。这些特性被确认为源于多巴胺的庞大性质使 GG 的螺旋结构变松。此外,多巴胺的亲水性特征被确认为通过促进溶胀能力和细胞附着,为细胞生长创造了更有利的微环境。当多巴胺的亲水性与多巴胺接枝水凝胶的较小多孔结构所带来的更大比表面积相结合时,这种改善的生物相容性变得更加明显。这可以从制备的水凝胶的活/死染色图像中看出。同时,非离子交联的 DFG(DG)水凝胶在培养 28 天后获得的免疫荧光染色图像中表现出最低的蛋白质表达,表明其具有最高的降解性和相关的细胞释放能力。这种趋势也可以从实时聚合酶链反应(RT-PCR)分析获得的基因表达数据中看出。RT-PCR 分析还表明,DG 水凝胶载体可以上调 RPE 的视觉功能相关基因。总的来说,DG 水凝胶系统证明了其作为 RPE 细胞载体的可行性,以及作为改善视觉功能的手段的潜力。