Cruise G M, Hegre O D, Scharp D S, Hubbell J A
Division of Chemistry and Chemical Engineering, California Institute of Technology, Mail Stop 210-41, Pasadena, California 91125, USA.
Biotechnol Bioeng. 1998 Mar 20;57(6):655-65. doi: 10.1002/(sici)1097-0290(19980320)57:6<655::aid-bit3>3.0.co;2-k.
A method has been defined to interfacially photopolymerize poly(ethylene glycol) diacrylates (PEG diacrylates) to form a crosslinked hydrogel membrane upon the surfaces of porcine islets of Langerhans to serve as an immune barrier for allo- and xenotransplantation. A sensitivity study of six key parameters in the interfacial photopolymerization process was performed to aid in determination of the optimal encapsulation conditions, leading to the most uniform hydrogel membranes and viable islets. The key parameters included the concentrations of the components of the initiation scheme, namely eosin Y, triethanolamine, and 1-vinyl 2-pyrrolidinone. Other parameters investigated included the duration and flux of laser irradiation and the PEG diacrylate molecular weight. Each parameter was doubled and halved from the standard conditions used in the encapsulation process while holding all the remaining parameters at the standard conditions. The effects of changing each parameter on islet viability, encapsulation efficiency, and gel thickness were quantified. Islet viability was sensitive to the duration of laser illumination, viability significantly increasing as the duration was reduced. Encapsulation efficiency was sensitive to the concentrations of eosin Y, triethanolamine, and 1-vinyl 2-pyrrolidinone, to the laser flux, and to the PEG diacrylate molecular weight. Increasing the concentration of eosin Y significantly improved the encapsulation efficiency, while decreasing the concentration of 1-vinyl 2-pyrrolidinone and increasing the concentration of triethanolamine had the greatest effects in significantly reducing the encapsulation efficiency. Gel thickness was sensitive to the concentrations of triethanolamine and 1-vinyl 2-pyrrolidinone, to the duration of laser illumination, and to the PEG diacrylate molecular weight. Increasing the PEG diacrylate molecular weight significantly increased the gel thickness, while decreasing the concentration of 1-vinyl 2-pyrrolidinone and increasing the concentration of triethanolamine had the greatest effects in significantly reducing the gel thickness. From this sensitivity study, conditions were determined to encapsulate porcine islets, resulting in greater than 90% islet viability and greater than 90% encapsulation efficiency.
已定义一种方法,通过界面光聚合聚乙二醇二丙烯酸酯(PEG二丙烯酸酯),在猪胰岛表面形成交联水凝胶膜,作为同种异体和异种移植的免疫屏障。对界面光聚合过程中的六个关键参数进行了敏感性研究,以帮助确定最佳封装条件,从而获得最均匀的水凝胶膜和存活的胰岛。关键参数包括引发体系各组分的浓度,即曙红Y、三乙醇胺和1-乙烯基-2-吡咯烷酮。研究的其他参数包括激光照射的持续时间和通量以及PEG二丙烯酸酯的分子量。在将所有其余参数保持在封装过程中使用的标准条件的同时,每个参数从标准条件加倍和减半。量化了改变每个参数对胰岛活力、封装效率和凝胶厚度的影响。胰岛活力对激光照射持续时间敏感,随着持续时间的减少,活力显著增加。封装效率对曙红Y、三乙醇胺和1-乙烯基-2-吡咯烷酮的浓度、激光通量以及PEG二丙烯酸酯的分子量敏感。增加曙红Y的浓度可显著提高封装效率,而降低1-乙烯基-2-吡咯烷酮的浓度和增加三乙醇胺的浓度对显著降低封装效率的影响最大。凝胶厚度对三乙醇胺和1-乙烯基-2-吡咯烷酮的浓度、激光照射持续时间以及PEG二丙烯酸酯的分子量敏感。增加PEG二丙烯酸酯的分子量可显著增加凝胶厚度,而降低1-乙烯基-2-吡咯烷酮的浓度和增加三乙醇胺的浓度对显著降低凝胶厚度的影响最大。通过这项敏感性研究,确定了封装猪胰岛的条件,从而使胰岛活力大于90%,封装效率大于90%。