傷口癒合的天然支架需具有可調機械性質和優良的生物相容性，在研究中，生物工程皮膚敷料，如海綿和膜，不能滿足傷口的幾何形狀，並提供濕潤，我們通過不同成膠溫度及曝光時間控制交聯程度，製造出具有可調的機械性能、微觀結構、交聯程度和降解速率的甲基丙烯酸酰基明膠 (Gelatin-methacryloyl (GelMA)) 水凝膠，透過不同交聯程度形成具有不同微結構的GelMA水凝膠，填入小鼠傷口中進行一階段治療，或是利用親水性水膠體敷料Duoderm(Positive control,PC)治療3天後，再放上GelMA，包裹間葉系幹細胞(mesenchymal stem cells,MSCs)的GelMA，或是同時包裹人臍靜脈內皮細胞(Human umbilical endothelial cells, HUVECs) 及間葉系幹細胞(mesenchymal stem cells,MSCs)，進行兩階段治療，28天後取下傷口，分析傷口部位上皮層、毛囊(真皮)層、皮下的脂肪層及肌肉層的覆蓋率，以及真皮層和肉芽組織厚度，探討不同水凝膠療法對皮膚再生的影響，探討血管新生對皮膚再生的影響；透過材料特性的機械性質、降解速率、微結構、水蒸發速率、膨潤比，比較對傷口癒合的影響，實驗結果顯示出要完整的再生無疤痕皮膚，需選用有較慢降解速率的水凝膠搭配人臍靜脈內皮細胞及間葉系幹細胞進行兩階段治療。透過這些探討，我們可以有效將水凝膠的物理化學特性應用在不同的材料系統上，未來可根據不同受傷程度的傷口需求而精準地控制皮膚的再生。

Engineering natural scaffolds with tunable mechanical properties, suitable biodegradation and great biocompatibility is highly desirable for wound healing. In previous study, bioengineering skin substitutes, like sponge and membrane, can’t totally cover the geometries of the wound and provide the wet physiological environment. In this study, we developed sequentially-crosslinked methacrylated GelMA hydrogels with tunable mechanical properties, degradation and microstructure by controlling molecules assembles, crosslinking degrees and crosslinked sites. GelMA hydrogels with different microstructures formed by different crosslinked sites and temperatures were filled into wounds for one-stage treatment, or treated with hydrophilic hydrocolloid dressing Duoderm (Positive control, PC) for 3 days, then put on GelMA, which encapsulated mesenchymal stem cells (MSCs), or simultaneously encapsulated human umbilical endothelial cells (HUVECs) and MSCs for two-stage treatment. After 28 days, the coverage of the epithelial layer, hair follicle (dermis) layer, subcutaneous fat layer and muscle layer of the wound site, as well as the thickness of the dermis layer and granulation tissuewere calculated to understand the effects of GelMA hydrogels with various conditions and therapies on skin regeneration. To understand how to regnerate the scar-free skin, the histological examination were performed after 7 days of treatment. The effects of mechanical properties, degradation rate, microstructure, water evaporation rate, swelling ratio of GelMA hydrogels were discussed. To completely regenerate scar-free skin, the hydrogel with a slower rate of degradation encapsulated HUVECs and MSCs were used in the two stages treatment. Through turning physico-chemical properties of GelMA hydrogels, we could presicely and effectively regenerate scar-free skin for various wounds.

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