巯基-烯光点击胶原-PEG水凝胶：水溶性光引发剂对细胞活力、凝胶动力学和流变学性质的影响
Thiol-ene photo-click collagen-PEG hydrogels: impact of water-soluble photoinitiators on cell viability, gelation kinetics and rheological properties
摘要：
使用硫醇官能化的I型胶原和8-臂聚（乙二醇）己烯封端的（PEG-NB）通过逐步生长聚合制备硫醇-烯光点击水凝胶，作为潜在的可注射再生装置。I型胶原通过与2-亚氨基硫杂环戊烷（2 IT）的开环反应进行硫醇官能化，其中通过2，4，6-三硝基苯磺酸（TNBS）比色测定和圆二色性（CD）记录了高达80 Abs%的官能化和90 RPN%的三螺旋保留。改变光引发剂的类型，即2-羟基-1-[4-（2-羟基乙氧基）苯基]-2-甲基-1-丙酮（I2959）或苯基-2，4，6-三甲基苯甲酰基次膦酸锂（I2959）和浓度，以确保最小的光引发剂诱导的细胞毒性，并使胶原蛋白-PEG混合物能够形成硫醇-烯网络。G292细胞在补充光引发剂的培养基中培养24小时后的活力在很大程度上受光引发剂浓度的影响，与补充LAP的培养基相比，观察到补充I2959的培养基诱导更高的毒性反应。与体外研究一致，使用选定的光引发剂浓度来制备硫醇-烯光点击水凝胶。甘精胰岛素动力学被证明在很大程度上受特定光引发剂的影响，含LAP的硫醇-烯混合物导致相对于含I2959的混合物显著减少的完全凝胶化时间。除了特定的光引发剂之外，光引发剂浓度是调节水凝胶储能模量（G '）的关键，由此在用0.5%（w/v）水凝胶制备的样品中观察到与0.1%（w/v）水凝胶相比15倍的G'增加。本研究建立的光引发剂-凝胶化-细胞毒性关系将有助于再生医学正交胶原基微生态位的设计。
英文摘要：
Thiol-ene photo-click hydrogels were prepared via step-growth polymerisation using thiol-functionalised type-I collagen and 8-arm poly(ethylene glycol) norbornene terminated (PEG-NB), as a potential injectable regenerative device. Type-I collagen was thiol-functionalised by a ring opening reaction with 2-iminothiolane (2IT), whereby up to 80 Abs% functionalisation and 90 RPN% triple helical preservation were recorded via 2,4,6-Trinitrobenzenesulfonic acid (TNBS) colorimetric assay and circular dichroism (CD). Type, i.e. 2-Hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methyl-1-propanone (I2959) or lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), and concentration of photoinitiator were varied to ensure minimal photoinitiator-induced cytotoxicity and to enable thiol-ene network formation of collagen-PEG mixtures. The viability of G292 cells following 24 h culture in photoinitiator-supplemented media was largely affected by the photoinitiator concentration, with I2959-supplemented media observed to induce higher toxic response compared to LAP-supplemented media. In line with the in vitro study, selected photoinitiator concentrations were used to prepare thiol-ene photo-click hydrogels. Gelation kinetics proved to be largely affected by the specific photoinitiator, with LAP-containing thiol-ene mixtures leading to significantly reduced complete gelation time with respect to I2959-containing mixtures. Other than the specific photoinitiator, the photoinitiator concentration was key to adjust hydrogel storage modulus (G'), whereby 15-fold G' increase was observed in samples prepared with 0.5% (w/v) compared to 0.1% (w/v) LAP. The photoinitiator-gelation-cytotoxicity relationships established in this study will be instrumental to the design of orthogonal collagen-based niches for regenerative medicine.
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