以聚甲基丙烯酸甲酯之微針陣列進行細胞傳遞研究

Abstract

細胞療法是在體外無菌環境下培養細胞後，將細胞取下並且傳遞到體內，恢復受損或缺失的細胞及組織以達到療效。過去研究中是以注射方式將細胞傳遞到體內，然而注射的方式會有幾個缺點，像是會造成患者疼痛感、感染風險、注射時必須由受過訓練人員操作、出血及形成血腫的可能性及會造成細胞迅速擴散，生物利用率低等。因此在本研究中，我們利用近期相當流行的微針陣列當作細胞的傳遞載體，其較小的尺寸，使皮下傷口較小，並且不會觸及皮下神經可以減輕患者疼痛感、不需要透過訓練人員協助操作、不易造成出血及降低感染風險等優點。此外，使用聚甲基丙烯酸甲酯作為微針的材料，其具有良好的機械強度、極佳的生物相容性、性能穩定耐用且價格相當低廉。因此本研究期望使用聚甲基丙烯酸甲酯製成的微針陣列用來傳遞細胞至皮下，以恢復患者缺失或受損的細胞。 在本研究中，使用聚二甲基矽氧烷製成的微針模具來製作聚甲基丙烯酸甲酯微針陣列。利用MTT assay觀察細胞的存活率以做材料的生物相容性評估。利用Live/Dead Staining觀察細胞在微針陣列上的貼附及增生情況，及利用電子顯微鏡觀察材料的表面結構及細胞貼附情形。利用膠原蛋白、海藻酸鈉及去細胞的新生鼠皮膚作為細胞轉移的材料，來確認細胞是否會成功從微針陣列上轉移至他處。 結果顯示聚甲基丙烯酸甲酯微針陣列具有良好的生物相容性，且細胞可以在其上貼附及增生。另外，培養在微針上的細胞也可以成功轉移至膠原蛋白、海藻酸鈉及去細胞的新生鼠皮膚中。因此，微針陣列搭配細胞療法是一個新穎且極具潛力的治療方式。

Cell therapy is a kind of treatment method that delivers cells which are cultured in vitro into the human body in order to recover missing or damaged cells. In the past, cells are delivered by injection, but this way may make patients painful, have the possibility for patients to get infected, need trained personnel to operate and have the possibility of forming hematoma or bleeding. In this study, we used microneedle array as a carrier for cells delivering. With its small size, the subcutaneous wound is smaller and it will not touch nerves under the skin, so improvement of patients’ comfort can be easily achieved. Besides, using microneedle array will not require trained personnel to assist operating. Finally, it will reduce the risk of infection and hematoma formation. We used polymethyl-methacrylate (PMMA) as the material for fabricating microneedle array due to its adequate mechanical strength, excellent biocompatibility, stability and lower price. We are looking forward to using microneedle array made of PMMA to act as a carrier for cells delivering in order to treat missing or damaged cells in the human body. In this study, we used polydimethylsiloxane(PDMS) mold for fabricating PMMA microneedle array. Biocompatibility and cell viability were evaluated by MTT assay. Cell adhesion and proliferation on microneedle array were examined by Live/Dead Staining and SEM. Moreover, we used collagen, alginate and acellular skin as a delivered material to observe whether the cells can deliver from microneedle array to another place. The results show that PMMA microneedle array has good biocompatibility and cells can attach and grow on it. Besides, cells on the microneedle array can successfully deliver to collagen, alginate and acellular skin. Therefore, microneedle array with cell therapy is a new and highly potential treatment method.