Sukhanova T V, Artiukhov A A, Prudchenko I A, Golunova A S, Seminikhina M A, Shtil'man M I, Markvicheva E A
Biomed Khim. 2013 Jan-Feb;59(1):65-75. doi: 10.18097/pbmc20135901065.
The aim of the study was to entrap delta-sleep inducing peptide (DSIP) in cross-linked poly(vinyl alcohol)-based hydrogels of different structures and to evaluate peptide release kinetics from these hydrogels using an in vitro model. Isotropic and macroporous hydrogels on the basis of poly(vinyl alcohol) acrylic derivative (Acr-PVA) as well as macroporous hydogels containing epoxy groups which were synthesized by copolymerization of this monomer with glycidyl methacrylate. The isotropic hydrogels were fabricated at positive temperatures while the macroporous hydrogels (cryogels) were prepared at the temperatures below zero. The peptide was entrapped into macroporous modified PVA hydrogels by addition of a peptide solution on previously fabricated matrices, while into PVA-GMA hydrogels containing epoxy groups peptide immobilization was carried out by incubation of hydrogel matrices in the peptide solution. In the case of isotropic hydrogels the peptide was added into the polymer mixture at a hydrogel formation reaction. The peptide release kinetics was studied by incubation of hydrogels in PBS (pH 7.4), in physiological solution (0.9% NaCl) and in water. DSIP concentration in supernatants was determined by phase-reverse HPLC. DSIP release from the macroporous PVA hydrogel after 30 min incubation was 74, 70 and 64% in water, PBS and 0.9% NaCl, relatively, and it was completed in 3 hs. From the isotropic hydrogel the release neither peptide nor products of its degradation was not observed even after 48 hs of incubation. For freshly prepared hydrogel the release kinetics was as follows: 27 and 78% in 30 and 33 hs, relatively. In the case of the lyophilized hydrogel samples the peptide release was 63% in 30 min incubation while drying patterns at room temperature for 3 days resulted in significant peptide loss because its structure damage.
本研究的目的是将δ-睡眠诱导肽（DSIP）包封在不同结构的交联聚乙烯醇基水凝胶中，并使用体外模型评估该肽从这些水凝胶中的释放动力学。基于聚乙烯醇丙烯酸衍生物（Acr-PVA）的各向同性和大孔水凝胶，以及通过该单体与甲基丙烯酸缩水甘油酯共聚合成的含环氧基团的大孔水凝胶。各向同性水凝胶在正温度下制备，而大孔水凝胶（冷冻凝胶）在低于零的温度下制备。通过在预先制备的基质上添加肽溶液，将肽包封到大孔改性PVA水凝胶中，而通过将水凝胶基质在肽溶液中孵育，将肽固定到含环氧基团的PVA-GMA水凝胶中。对于各向同性水凝胶，在水凝胶形成反应时将肽添加到聚合物混合物中。通过将水凝胶在PBS（pH 7.4）、生理溶液（0.9% NaCl）和水中孵育来研究肽的释放动力学。通过反相HPLC测定上清液中DSIP的浓度。在水中、PBS和0.9% NaCl中孵育30分钟后，大孔PVA水凝胶中DSIP的释放率分别为74%、70%和64%，并在3小时内完成。即使孵育48小时后，也未观察到各向同性水凝胶释放肽或其降解产物。对于新鲜制备的水凝胶，释放动力学如下：在30小时和33小时时分别为27%和78%。对于冻干的水凝胶样品，在孵育30分钟时肽释放率为63%，而在室温下干燥3天导致肽大量损失，因为其结构受损。
