目的:基于水凝胶的玻璃体替代品具有克服目前临床使用的内安瓿的局限性的潜力。为了进入临床试验,本研究旨在(I)将透明质酸水凝胶的材料合成转移到常规中,(II)根据现行医疗器械法规(MDR/ISO标准)评估玻璃体替代品的性能。
方法:玻璃体代用品的多步骤制造过程,包括用甲基丙烯酸缩水甘油酯修饰透明质酸,与N-乙烯基吡咯烷酮的光共聚,和连续的水凝胶纯化,是在实验室条件下开发的,用1H-NMR表征,FT-IR和UV/Vis光谱和HPLC,并转向考虑GMP标准的药品生产环境。将基于透明质酸的水凝胶的光学和粘弹性特性与提取的人玻璃体和硅油的光学和粘弹性特性进行了比较。水凝胶对代谢活性的影响,成纤维细胞的增殖和凋亡(MRC-5,BJ,L929),通过HET-CAM测定研究了视网膜色素上皮(ARPE-19,hiPSC衍生的RPE)和感光细胞(661W)及其粘膜耐受性。
结果:具有合适纯度的基于透明质酸的水凝胶,不育,高透明度(>90%),适当的折射率(1.3365)和粘弹性(G'>G")在受控的工艺条件下以标准化的方式制备。代谢活动,各种细胞类型以及卵脉络膜的增殖和凋亡不受基于透明质酸的玻璃体替代物的影响,证明了它们的生物相容性。
结论:本研究表明,透明质酸水凝胶的关键合成步骤成功地转移到常规,符合GMP的生产工艺,同时实现光学和粘弹性,临床用作玻璃体替代品所需的生物相容性和纯度。
OBJECTIVE: Hydrogel-based vitreous substitutes have the potential to overcome the limitations of current clinically used endotamponades. With the goal of entering clinical trials, the present study aimed to (I) transfer the material synthesis of hyaluronic acid-based hydrogels into a routine, pharmaceutical-appropriate production and (II) evaluate the properties of the vitreous substitutes in terms of the current regulations for medical devices (MDR/ISO standards).
METHODS: The multistep manufacturing process of the vitreous substitutes, including the modification of hyaluronic acid with glycidyl methacrylate, photocopolymerization with N-vinylpyrrolidone, and successive hydrogel purification, was developed under laboratory conditions, characterized using 1 H-NMR, FT-IR and UV/Vis spectroscopies and HPLC, and transferred towards a pharmaceutical production environment considering GMP standards. The optical and viscoelastic characteristics of the hyaluronic acid-based hydrogels were compared with those of extracted human vitreous and silicone oil. The effect of the hydrogels on the metabolic activity, proliferation and apoptosis of fibroblast (MRC-5, BJ, L929), retinal pigment epithelial (ARPE-19, hiPSC-derived RPE) and photoreceptor cells (661W) was studied as well as their mucosal tolerance via a HET-CAM assay.
RESULTS: Hyaluronic acid-based hydrogels having a suitable purity, sterility, high transparency (>90%), appropriate refractive index (1.3365) and viscoelasticity (G\' > G″) were prepared in a standardized manner under controlled process conditions. The metabolic activity, proliferation and apoptosis of various cell types as well as egg choroid were unaffected by the hyaluronic acid-based vitreous substitutes, demonstrating their biocompatibility.
CONCLUSIONS: The present study demonstrates the successful transferability of the crucial synthesis steps of hyaluronic acid-based hydrogels into a routine, GMP-compliant production process while achieving the optical and viscoelastic properties, biocompatibility and purity required for their clinical use as vitreous substitutes.