冷凍微針貼片於經皮細胞治療之應用

Abstract

用於疾病治療的細胞療法可以透過簡易、有效與安全的經皮傳遞具治療效果細胞的技術來完成，傳統透過皮下或靜脈注射進行細胞傳遞會受患者依從性所影響，不同批次的細胞品質也不盡相同，且細胞的施打也需要訓練有素的醫療人員進行，如此不只使療效受到影響，也可能造成醫療資源吃緊。本篇研究使用特殊配製的冷凍保存液所製成冷凍微針貼片，裝載人類羊水幹細胞用於促進毛髮生長；這種冷凍微針是將懸浮細胞加入由透明質酸鈉、二甲基亞石碸與海藻糖所配製成的冷凍保存液，並透過漸進式冷凍的方式使細胞在良好的條件下被保存，同時也能使裝填於聚二甲基矽氧烷（Polydimethylsiloxane, PDMS）模中的冷凍保存液凍結成為微針的形狀。 當冷凍微針刺穿大鼠皮膚並當微針溶解後，裝載其中的人類羊水幹細胞便會擴散而出，與其他部位相比，施打含有人類羊水幹細胞冷凍微針部位的毛髮生長速度較快，不僅如此，取人類羊水幹細胞的條件培養基（Conditioned medium）製成之冷凍微針，在施打於大鼠後也與施打人類羊水幹細胞冷凍微針有相同的效果；另外，我們也製作了裝載表現紅色螢光蛋白的豬骨髓間質幹細胞（Porcine Bone Marrow Mesenchymal Stem Cell）的冷凍微針，並將之使用於大鼠與去細胞牛眼角膜之上，觀察使用後不同時間的螢光強度與分布狀態，用以模擬細胞進入體內的生長與擴散情形，實驗顯示冷凍微針可以有效地將細胞傳送到目標組織，並均勻地分散於組織中生長，此系統未來很有潛力應用於細胞治療的領域。

Cell therapy for disease treatment can be completed through simple, effective, and safe transdermal delivery of therapeutic cells. The patient's compliance will affect the traditional cell delivery through subcutaneous or intravenous injection. The cell quality of different batches is different, and the injection of cells also needs trained medical personnel. This will affect the curative effect and cause a shortage of medical resources. In this study, we used a unique recipe of cryopreservation to make cryomicroneedle patch loaded with human amniotic fluid stem cells (hAFSCs) to promote hair growth. The cryomicroneedle is to add the suspended cells into the cryopreservation solution prepared with sodium hyaluronate, dimethyl sulfoxide, and trehalose and preserve the cells under ideal conditions through progressive freezing. Simultaneously, it can also freeze the solution filled in the polydimethylsiloxane (PDMS) mold into the shape of microneedles. When the cryomicroneedle pierces the rat skin and dissolves, the human amniotic fluid stem cells will spread out. Compared with other body parts without cryomicroneedle administration, the hair grew faster when applied with the cryomicroneedle loaded with hAFSCs. In addition, the cryomicroneedle made from the conditioned medium of h hAFSCs also has the same effect as that of h hAFSCs cryomicroneedle in rats. Moreover, we also fabricate cryomicroneedle loaded with porcine bone marrow mesenchymal stem cells (PBM-MSCs) that expressed red fluorescent protein. And applied the cryomicroneedle patches loaded with PBM-MSCs to rat and decellularized bovine cornea to observe the fluorescence intensity and distribution at different times after administration to simulate and confirm the growth and diffusion of cells.