Chou Hung-Da, Chen Chung-An, Liu Hao-Yu, Liu Shih-Jung, Lai Po-Liang, Wu Wei-Chi, Hwang Yih-Shiou, Chen Kuan-Jen, Tsai Tsung-Ting, Lai Chi-Chun
Department of Ophthalmology, Chang Gung Memorial Hospital, Linkou Main Branch, Taoyuan 333, Taiwan.
Department of Orthopaedic Surgery, Spine Section and Bone and Joint Research Center, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan.
ACS Omega. 2024 Oct 1;9(41):42147-42158. doi: 10.1021/acsomega.4c02842. eCollection 2024 Oct 15.
The current surgical adjunctive for vitreoretinal surgeries fails to provide an adequate 3D structure for cellular regeneration. A one-pot synthesis of gelatin-methacryloyl (GelMA) followed by modification with 4-carboxyphenylboronic acid (4-CPBA) was performed to fabricate 4-CPBA-modified GelMA (4CPBA@GelMA), a gelatin-based hydrogel. 4CPBA@GelMA was photo-cross-linked by 405 nm violet light and examined using nuclear magnetic resonance (NMR), Fourier-transform infrared spectrometry (FTIR), scanning electron microscopy (SEM), and rheometry. In vitro biocompatibility was examined by Müller cell proliferation assays exposed to 4CPBA@GelMA and violet light. In vivo retinal biocompatibility was evaluated by electroretinography of rat eyes that were exposed to intravitreally injected and photo-cross-linked 4CPBA@GelMA at days 3, 7, 14, and 28 post-injection. Following electroretinography, histology and immunohistochemistry were performed on the retinas. The NMR results indicated amidation of GelMA by 4-CPBA, and FTIR confirmed the presence of the CPBA ring in 4CPBA@GelMA samples. SEM revealed that 4CPBA@GelMA had significantly larger pores than GelMA (56.9 ± 9.5 vs 35.1 ± 2.8 μm; < 0.001). Rheological findings showed that, unlike GelMA and gelatin, 4CPBA@GelMA has Newtonian fluid properties at room temperature. Exposure to 4CPBA@GelMA did not significantly affect Müller cell viability in a proliferation assay; moreover, electroretinography findings indicated normal waveforms and implicit times, and histology and immunohistochemistry examinations revealed no significant changes. In this study, we established the high retinal compatibility of 4CPBA@GelMA. The low viscosity of 4CPBA@GelMA is ideal for injection via small-gauge needles, and the larger pore size and three-dimensional network both potentiate cellular migration and growth. These features made 4CPBA@GelMA a candidate for vitreoretinal surgical adjunctive that might promote retinal regeneration.
目前用于玻璃体视网膜手术的手术辅助材料无法为细胞再生提供足够的三维结构。进行了一锅法合成甲基丙烯酰化明胶（GelMA），随后用4-羧基苯硼酸（4-CPBA）进行改性，以制备基于明胶的水凝胶4-CPBA改性GelMA（4CPBA@GelMA）。4CPBA@GelMA通过405nm紫光进行光交联，并使用核磁共振（NMR）、傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）和流变学进行检测。通过暴露于4CPBA@GelMA和紫光的 Müller 细胞增殖试验来检测体外生物相容性。通过对注射后第3、7、14和28天玻璃体内注射并光交联的4CPBA@GelMA的大鼠眼睛进行视网膜电图检查来评估体内视网膜生物相容性。视网膜电图检查后，对视网膜进行组织学和免疫组织化学检查。NMR结果表明4-CPBA对GelMA进行了酰胺化，FTIR证实了4CPBA@GelMA样品中存在CPBA环。SEM显示4CPBA@GelMA的孔隙明显大于GelMA（56.9±9.5对35.1±2.8μm；<0.001）。流变学结果表明，与GelMA和明胶不同，4CPBA@GelMA在室温下具有牛顿流体特性。在增殖试验中，暴露于4CPBA@GelMA对Müller细胞活力没有显著影响；此外，视网膜电图结果表明波形和隐含时间正常，组织学和免疫组织化学检查未发现显著变化。在本研究中，我们确定了4CPBA@GelMA具有高视网膜相容性。4CPBA@GelMA的低粘度非常适合通过小口径针头注射，较大的孔径和三维网络都有利于细胞迁移和生长。这些特性使4CPBA@GelMA成为可能促进视网膜再生的玻璃体视网膜手术辅助材料的候选者。
