Institute for Technical and Macromolecular Chemistry, University of Hamburg , Bundesstrasse 45, D-20146, Hamburg, Germany.
Biomacromolecules. 2017 Nov 13;18(11):3714-3725. doi: 10.1021/acs.biomac.7b00382. Epub 2017 Oct 9.
Thermoresponsive and active functional fiber mats were prepared from random copolymer of poly(pentafluorophenyl acrylate-co-N-isopropylacrylamide) (P(PFPA-co-NIPAM)), which was synthesized by a controlled radical polymerization process based on reversible addition-fragmentation chain transfer (RAFT). As reactive sites, pentafluorophenyl ester (PFP) groups were incorporated in the copolymer to allow for a multiple post-polymerization modification. UV-cross-linkable moieties were first introduced by partially reacting PFP groups in the copolymer with ortho-nitrobenzyl (ONB)-protected diamine. Electrospinning the resulting ONB-containing P(PFPA-co-NIPAM), followed by UV-induced cross-linking, yielded stable cross-linked thermoresponsive PNIPAM-based fiber mats. The remaining PFP active groups on the surface of copolymer fiber mats allowed for further conjugation with an H-Gly-Arg-Gly-Asp-Ser-OH (GRGDS) peptide, a well-known cell adhesive peptide sequence that was selected as a model in order to promote cell growth. At 37 °C, fibroblast cells were found to attach, spread, and proliferate well on the GRGDS-immobilized cross-linked (CL) fiber mat, as opposed to those on the GRGDS-immobilized un-cross-linked (UCL) fiber mat. By decreasing the temperature down to 20 °C, i.e. below the lower critical solution temperature (LCST) of thermoresponsive PNIPAM, cultured cells could easily be released from both GRGDS-immobilized CL and UCL fiber mats, whereas no cells were detached from tissue culture polystyrene (TCPS). These results suggest that the thermosensitive and active functional fiber mat obtained in this research represent an attractive and versatile platform for cultured cell recovery, which is beneficial for tissue engineering applications.
温敏性和功能化纤维垫由聚(五氟苯丙烯酸酯-co-N-异丙基丙烯酰胺)(P(PFPA-co-NIPAM))无规共聚物制备而成,该共聚物通过基于可逆加成-断裂链转移(RAFT)的可控自由基聚合过程合成。作为反应性基团,五氟苯酯(PFP)基团被引入共聚物中,以允许进行多次聚合后修饰。首先通过将共聚物中的部分 PFP 基团与邻硝基苄基(ONB)保护的二胺反应,引入可光交联的部分。将得到的含 ONB 的 P(PFPA-co-NIPAM)进行静电纺丝,然后进行 UV 交联,得到稳定的交联温敏性 PNIPAM 基纤维垫。共聚物纤维垫表面上剩余的 PFP 活性基团允许进一步与 H-Gly-Arg-Gly-Asp-Ser-OH(GRGDS)肽缀合,GRGDS 肽是一种众所周知的细胞黏附肽序列,被选为模型以促进细胞生长。在 37°C 下,发现成纤维细胞在 GRGDS 固定的交联(CL)纤维垫上很好地附着、展开和增殖,而在 GRGDS 固定的未交联(UCL)纤维垫上则不然。当温度降低至 20°C 以下,即低于温敏性 PNIPAM 的下临界溶液温度(LCST)时,培养的细胞可以很容易地从 GRGDS 固定的 CL 和 UCL 纤维垫上释放出来,而从组织培养聚苯乙烯(TCPS)上则没有细胞脱落。这些结果表明,本研究中获得的温敏性和功能化纤维垫代表了一种有吸引力和多功能的培养细胞回收平台,这有利于组织工程应用。