智能型溫感性雙離子體水凝膠之製備及其細胞吸脫附性質之轉換控制

Preparation of intelligent thermoresponsive zwitterionic hydrogels and its switchable control of cell attachment/detachment

潘嘉鴻(Chia-Hung Pan)

指導教授 ： 張雍

中原大學/工學院/化學工程研究所/碩士(2011年)

https://doi.org/10.6840/cycu201100562

摘要

本研究以電中性之雙離子單體與帶電性之離子單體來修飾溫度敏感性高分子聚異丙基丙烯胺(poly(N-isopropyl acrylamide), PNIPAAm)，擬設計出兼具高細胞貼附性與快速細胞脫附性之溫感型智能水凝膠，未來可應用於細胞層片組織工程(cell sheet engineering)方面的應用。因此，本研究欲以添加帶電官能基來增加材料與細胞間的靜電吸引力，期提高細胞於膜材之貼附性，並以電中性之雙離子官能基來增加材料與細胞間的水層轉移效應，來加強細胞於膜材之脫附性，細胞吸脫附性質將以溫度進行切換控制。
本論文的第一部分為探討分別加入單一成分之電中性雙離子單體(sulfobetaine methacrylate, SBMA)、帶負電離子單體(11-mercaptoundecylsulfonic acid, SA)或帶正電離子單體 (11-mercapto-N,N,N-trimethylammonium chloride, TMA) 於PNIPAAm水凝膠中，精準控制所添加單體與NIPAAm的成分比例，並探討其細胞吸脫附性質。實驗過程中，以37 oC來進行細胞培養與貼附觀察，並以20 oC來進行細胞脫附的動態觀察。實驗結果顯示，細胞的吸附與脫附行為會同時受到水凝膠表面之親水性與電荷量影響，當PNIPAAm水凝膠加入帶電成分SA或TMA時，會使得帶電之PNIPAAm水凝膠比電中性PNIPAAm水凝膠吸引更多細胞貼附，但會延長細胞自材料表面脫附下來的時間，當帶電成分過高時，細胞無法產生脫附行為。
本論文的第二部分為探討加入電中性雙離子單體於帶正電之PNIPAAm水凝膠，在精準控制水凝膠中各成分比例，並探討其細胞吸脫附性質。實驗結果顯示，當電中性雙離子成分SBMA加入帶正電之水凝膠中，不僅可保有高度的細胞貼附量，且可加速細胞之脫附量。研究結果指出，當PNIPAAm水凝膠含有10 mol%的正電TMA成分與20 mol%的雙離子SBMA成分時，其細胞貼附性與細胞脫附性會優越於100 mol%的PNIPAAm水凝膠。新的雙離子帶電型水凝膠更容易吸引細胞貼附，同時也具有加速細胞脫附的性質，可成為新一代兼具高細胞貼附性與快速細胞脫附性之溫感型智能水凝膠。

關鍵字

細胞貼附表面 ； 溫度敏感性 ； 快速細胞脫附性

並列摘要

This thesis describes a tunable cell-adhesive hydrogel of thermoresponsive poly(N-isopropyl acrylamide) (PNIPAAm) containing zwitterionic monomers and charged monomers. The designed hydrogel is expected to have two functions in high attachment and rapid detachment of cultured cells. It makes this new hydrogel appropriate for cell sheet engineering in the future. In this work, the functional groups of charged moieties in the PNIPAAm gels were used to enhance the cell attachment using electrostatic interaction of gel interfaces with cells. The incorporation of zwitterionic units into the PNIPAAm gels was expected to trigger cell detachment using the transformation of hydration layer. Temperature was used to control the switchable functions between cell attachment and cell detachment.

In the first part of this dissertation, zwitterionic monomer (sulfobetaine methacrylate, SBMA), anionic monomer (11-mercaptoundecylsulfonic acid, SA), or cationic monomer (11-mercapto-N,N,N-trimethylammonium chloride, TMA) were used to add into PNIPAAm gels with controlled compositions and study the effects on cell attachment at 37 oC and cell detachment at 20 oC. Results showed that the hydrophilicity and charged amounts on gel interfaces exhibit an obviously influence in cell attachment and detachment. The increase of charged amounts on gels resulted in more amounts of cell attachment and lower speed of cell detachment compared to the virgin gels.

In the second part of this dissertation, zwitterionic monomers were used to add into cationic PNIPAAm with controlled nonionic/ionic compositions and study the effects on cell attachment and cell detachment. Results showed that the cationic gels containing zwitterionic SBMA provide not only high amounts of cell attachment but also high speed of cell detachment. This work shows that the PNIPAAm hydrogel with an optimized composition of about 10 mol% cationic TMA and 20 mol% SBMA is presented cell capability to function the best switchable character of cell attachment and detachment. The excellent tunable cell-adhesive interface of new zwitterionic-cationic PNIPAAm hydrogels along with their thermoresponsive property suggests their potential for use in cell-culturing materials and cell sheet engineering.

並列關鍵字

cell-adhesive surface ； rapid cell detachment ； temperature-sensitive

參考文獻

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