可注射温敏性水凝胶用于三维细胞支架材料的分析-analysis of injectable thermosensitive hydrogel for three-dimensional cell scaffold materials.docxVIP

摘要摘要在本文中我们主要研究了两种温敏性水凝胶细胞支架材料。 首先，我们进行了温敏性的P(NIPAM．AAc)水凝胶用于HepG2细胞团的原位形成和释放的研究。细胞团的获得是通过器官打印的方法来构建功能性组织或器官的重要前提。P(NIPAM．AAc)凝胶颗粒在一定条件下加热到37℃可以收 缩成块状凝胶，冷却到室温后，块状凝胶发生溶胶化。HepG2细胞可以在 P(NIPAM．AAc)凝胶中生长。由于细胞和支架材料之间的相互作用较弱，因此 细胞间的相互作用使得细胞聚集进而形成细胞团。当温度冷却到室温时，凝胶 溶化，细胞团从凝胶中释放出来。释放过程只是温度的改变，没有引入化学物 质也没有使得细胞的生长条件发生剧烈变化，因此细胞及DNA并不受到损害。 并且释放过程中，细胞团的大小可以控制。PNIPAM水凝胶在临床应用的一个很大的局限就是不具备生物降解性。因 此在第二个工作中，我们利用聚己内酯(PCL)这种可降解高分子材料，首先 制备得到了PCL微球，接下来在其表面进行改性，接枝引入PNIPAM，最终得 到聚己内酯．聚N．异丙基丙烯酰胺(PCL-PNIPAM)核壳微球凝胶，该种材料具有可注射、温度敏感性且体积相转变温度接近人体温度、可原位凝胶、可生物降解等优点，适合用作可注射三维支架材料。实验结果证明，(PCL．PN删)核壳微球凝胶相转变温度为30℃左右，细胞在材料内能够保持一定的活性，经 过一段时间的培养，有较大的细胞团生成，证明该支架材料的生物相容性。关键词：温度敏感性 水凝胶可注射三维细胞支架材料 生物可降解AbstractAbstractIn present work,two kinds of thermosensitive hydrogels scaffolds have been fabricated．Firstly,we synthesized P(NIPAM-ARc)hydrogels．We presented here that thermoreversible hydrogels may be used for the fabrication of multicellular spheroids，which are the building blocks for further assembly to produce functionaltissues or organs using the organ printing approach．We showed that under properconditions the PNIPAM microgel dispersions solidify by simple heating to 37C， and they return to be liquid when cooled back to room temperature．HepG2 cells were seeded and cultured inside the in situ·formed hydrogels．They form multicellular spheroids because of the weak cell—substrate interactions．The resultantspheroids were released by simple cooling to room temperature．The materials usedhere are totally synthesized polymers and free from any biological contamination． No additive or harsh conditions，which may induce injury to cell or DNA are used． More importantly,the process is scalable，thus，may meet the requirement for large scale fabrication of cell spheroids．The lack of biodegradability had limited the clinical application of PNIPAMhydrogels as the injectable scaffold for 3D eell culture．Therefore．in our secondwork，we prepared the polycaprolactone nanospheres，which is biodegradable，and then sustained PCL-PNIPA