Tissue Culture Laboratory, Division of Implant Biology, Biomedical Technology Wing, Sree Chithra Tirunal Institute for Medical Sciences and Technology, Trivandrum, Kerala, India.
J Mater Sci Mater Med. 2010 May;21(5):1631-9. doi: 10.1007/s10856-010-3992-x. Epub 2010 Jan 22.
Scaffold free tissue constructs are preferred in tissue engineering as they overcome all the problems associated with scaffolds. Stimuli responsive polymers enable generation of scaffold free multilayered tissue constructs which would in turn reduce the use of biomaterials in vivo. In this study, we investigated cytocompatibility and thermoresponsiveness of a copolymer of N-Isopropylacrylamide and Methyl Methacrylate. Thermoresponsive surfaces were prepared by coating tissue culture polystyrene with the copolymer solution in isopropanol. Mammalian fibroblast cells (L929 cells) readily adhered on the copolymer. The viability and cellular activity was ensured through Neutral red staining, MTT assay, Tritiated thymidine uptake assay and Immunofluorescent staining for cytoskeletal organisation. Incubation under lower critical solution temperature of copolymer resulted in intact detachment of cells. To conclude, in-house synthesized cytocompatible smart culture substrate intended for tissue engineering was developed using a cost effective and simple technique. Moreover, presence of methyl methacrylate in the copolymer reduced the lower critical solution temperature facilitating extended in vitro manipulation time. As the copolymer is insoluble in water, the copolymer could be polymerised without additional crosslinkers.
无支架组织构建物在组织工程中是优选的，因为它们克服了与支架相关的所有问题。刺激响应性聚合物能够产生无支架的多层组织构建物，这反过来又会减少体内生物材料的使用。在这项研究中，我们研究了 N-异丙基丙烯酰胺和甲基丙烯酸甲酯的共聚物的细胞相容性和温敏性。通过将共聚物溶液涂覆在异丙醇中的组织培养聚苯乙烯上，制备温敏表面。哺乳动物成纤维细胞（L929 细胞）很容易附着在共聚物上。通过中性红染色、MTT 测定、氚标记胸腺嘧啶摄取测定和细胞骨架组织的免疫荧光染色，确保了细胞活力和细胞活性。在共聚物的较低临界溶液温度下孵育会导致细胞完整脱离。总之，使用成本效益高且简单的技术，开发了用于组织工程的合成细胞相容性智能培养底物。此外，共聚物中甲基丙烯酸甲酯的存在降低了较低临界溶液温度，从而延长了体外操作时间。由于共聚物不溶于水，因此可以在没有额外交联剂的情况下聚合共聚物。
