【摘要】：背景:有研究顯示脂肪干細(xì)胞參與燙傷后皮膚組織的修復(fù),但利用脂肪干細(xì)胞分泌物制備水凝膠,并治療皮膚燙傷的研究尚未見(jiàn)報(bào)道。目的:分析人脂肪干細(xì)胞條件培養(yǎng)基水凝膠外用治療小鼠皮膚燙傷模型的療效。方法:采用酶解聯(lián)合貼壁培養(yǎng)法,從脂肪組織培養(yǎng)獲得脂肪干細(xì)胞并進(jìn)行鑒定。取穩(wěn)定增殖期細(xì)胞,收集其條件培養(yǎng)基,加入殼聚糖、甘露醇、β-甘油磷酸鈉和透明質(zhì)酸等制備溫敏水凝膠,同時(shí)用新鮮脂肪干細(xì)胞完全培養(yǎng)基制備水凝膠。24只C57BL/6小鼠背部脫毛后用95℃鋁塊燙傷10 s快速建立3度皮膚燙傷模型,右側(cè)以脂肪干細(xì)胞條件培養(yǎng)基水凝膠每天涂抹2次;左側(cè)使用脂肪干細(xì)胞新鮮培養(yǎng)基水凝膠每天涂抹2次,連續(xù)涂抹7 d,觀察愈合時(shí)間與愈合進(jìn)程,計(jì)算愈合率,在燙傷后第4,14,28天取材進(jìn)行蘇木精-伊紅染色觀察組織病理學(xué)變化。結(jié)果與結(jié)論:①人脂肪間充質(zhì)干細(xì)胞為成纖維狀,增殖旺盛,平均倍增時(shí)間為55 h,可被誘導(dǎo)分化為成骨細(xì)胞、軟骨細(xì)胞和脂肪細(xì)胞,高表達(dá)CD29、CD44、CD90和CD105,低表達(dá)CD31和CD34,證明其符合間充質(zhì)干細(xì)胞的標(biāo)準(zhǔn);②用人脂肪間充質(zhì)干細(xì)胞條件培養(yǎng)基制備的溫敏水凝膠在4-20℃呈現(xiàn)為清涼透明的黏稠液體,放入37℃15 min變?yōu)榘牍虘B(tài)凝膠;③95℃鋁塊燙傷小鼠表皮、真皮及皮下脂肪和肌肉組織的正常結(jié)構(gòu)完全破壞,符合嚴(yán)重的3度燙傷標(biāo)準(zhǔn),傷口面積均大致為3 cm~2;④在修復(fù)過(guò)程中發(fā)現(xiàn),與新鮮培養(yǎng)基水凝膠對(duì)照組相比,脂肪干細(xì)胞條件培養(yǎng)基水凝膠組創(chuàng)口愈合時(shí)間短,傷口整潔,瘢痕增生少,創(chuàng)面組織結(jié)構(gòu)恢復(fù)好;⑤脂肪干細(xì)胞條件培養(yǎng)基水凝膠組炎癥浸潤(rùn)深度、炎癥浸潤(rùn)細(xì)胞密度、肉芽組織厚度、成纖維細(xì)胞密度、新生血管密度和表皮厚度等指標(biāo)顯著好于新鮮培養(yǎng)基水凝膠組,差異有顯著性意義(P0.05);⑥以上結(jié)果表明,人脂肪間充質(zhì)干細(xì)胞條件培養(yǎng)基水凝膠可有效促進(jìn)燙傷皮膚傷口愈合,提高再生皮膚質(zhì)量。
[Abstract]:Background: some studies have shown that adipose stem cells are involved in the repair of skin tissue after scald, but the preparation of hydrogel from adipose stem cell secretions and the treatment of skin scald have not been reported. Objective: to analyze the efficacy of human adipose stem cells (HSC) conditioned culture medium hydrogel in the treatment of mouse skin scald model. Methods: adipose stem cells were obtained from adipose tissue by enzyme hydrolysis combined with adherent culture. The thermosensitive hydrogel was prepared by adding chitosan, mannitol, sodium 尾-glycerophosphate and hyaluronic acid into the conditioned medium for stable proliferation of cells. At the same time, hydrogel was prepared by fresh adipose stem cell complete medium. 24 C57BL/6 mice were scalded with 95 鈩，

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