【摘要】：目的探討自體表皮細胞懸液移植技術用于全層皮膚缺損創(chuàng)面修復的適宜密度。方法取健康清潔級成年SD大鼠40只,雌雄不限,體質(zhì)量210~230 g;根據(jù)細胞移植密度不同,隨機分為高、中、低細胞密度及空白組(分別為A、B、C、D組,n=10)。取大鼠背部皮膚培養(yǎng)表皮細胞,并制作大鼠全層皮膚缺損創(chuàng)面抗攣縮模型。其中A、B、C組分別將0.2 mL密度為1×10~6、1×10~5、1×10~4個/cm~2的自體表皮細胞懸液移植至創(chuàng)面處,D組給予等量限制性角質(zhì)形成細胞無血清培養(yǎng)基;取成年Wistar大鼠背部皮膚制備同種異體皮,覆蓋各組創(chuàng)面。術后觀察大鼠存活情況,于術后7、14、21 d大體觀察同種異體皮成活、脫落及創(chuàng)面愈合情況,同種異體皮脫落后計算創(chuàng)面愈合率;21 d時取材行組織學及免疫組織化學染色,觀察創(chuàng)面修復情況。結(jié)果術后大鼠均存活至實驗完成。各組大鼠同種異體皮隨時間延長逐漸成活,干燥并開始脫痂;至21 d同種異體皮基本脫落后A、B組創(chuàng)面可見成片上皮,C組創(chuàng)面可見少量菲薄上皮,D組創(chuàng)面無上皮形成。術后21 d同種異體皮脫落后,A、B、C、D組創(chuàng)面愈合率分別為62.9%±9.6%、64.2%±9.1%、38.5%±5.7%、22.7%±5.5%,A、B組創(chuàng)面愈合率顯著高于C、D組(P0.05),C組高于D組(P0.05),A、B組間比較差異無統(tǒng)計學意義(P0.05)。組織學觀察示,A、B、C組愈合的創(chuàng)面上皮層可見鱗狀上皮細胞,A、B組表皮分層明顯,C組表皮層薄、可見炎性細胞浸潤,D組為肉芽組織。免疫組織化學染色觀察示,A、B、C組表皮-真皮連接層Ⅳ型膠原和Ⅶ型膠原表達呈陽性,D組無表皮層呈陰性;A、B組Ⅳ、Ⅶ型膠原表達陽性細胞百分比顯著高于C組(P0.05),A、B組間比較差異無統(tǒng)計學意義(P0.05)。結(jié)論自體表皮細胞懸液移植技術在大鼠全層皮膚缺損創(chuàng)面修復中可重構皮膚,1.0×10~5個/mL為創(chuàng)面修復的適宜移植密度。
[Abstract]:Objective to investigate the proper density of autologous epidermal cell suspension transplantation for full-scale skin defect repair. Methods Forty healthy clean grade adult SD rats, male and female, 210g / 230g, were randomly divided into high, middle and low cell density and blank group (n = 10). The skin of rat back was cultured with epidermal cells and the model of anti contracture was established. In group A, the suspension of autologous epidermal cells with a density of 0. 2 mL 1 脳 10 ~ 6 and 1 脳 10 ~ 5 / cm~2 was transplanted to the wound, and group D was given the same amount of serum-free medium for restricted keratinocytes. Allogeneic skin was prepared from the dorsal skin of adult Wistar rats to cover the wounds in each group. The survival of the rats was observed after 714 days. The survival rate of allogeneic skin and wound healing were observed on the 21st day after operation. The rate of wound healing was calculated after allogeneic skin exfoliation. The wound repair was observed by histological and immunohistochemical staining at 21 d. Results all the rats survived until the experiment was completed. The allogeneic skin in each group survived gradually with time, dried and began to unscab, and at 21 days after the abscission of the allogeneic skin, the lamellar epithelium was found in group A B, a small amount of thin epithelium was found in group C, and no epithelium was formed in group D. On the 21st day after operation, the healing rate of the wounds in the group A BX Con D was 62.9% 鹵9.6% and 64.2% 鹵9.1% respectively, and 38.5% 鹵5.7% 鹵5.5A, respectively, in the group A, the healing rate of the wounds was 22.7% 鹵5.5A, and the healing rate of the wounds in group A was 62.9% 鹵9.6% 鹵9.2% 鹵9.1%, respectively. The rate of wound healing in group B was significantly higher than that in group C (P0.05) (P0.05) in group), C and group D (P0.05), but there was no significant difference between group B and group A (P0.05). Histological observation showed that squamous epithelium cells could be seen in the epithelium of wound healing in group A BX C, epidermis in group A B was obviously stratified, epidermis in group C was thin and inflammatory cells were infiltrated, and granulation tissue was found in group D. Immunohistochemical staining showed that the expressions of type 鈪，

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