Yin Kai, Li Dawei, Shen Chuanan, Shang Yuru, Li Longzhu, Zhao Dongxu, Cheng Wenfeng
Department of Burns and Plastic Surgery, Beijing Jishuitan Hospital, Beijing, China.
Department of Burns and Plastic Surgery, The Fourth Medical Center of Chinese PLA General Hospital, Beijing, China,
Eur Surg Res. 2022;63(4):269-277. doi: 10.1159/000526305. Epub 2022 Aug 4.
The high mortality of patients with extensive deep burns is mainly attributed to the extensive burn wound and the scarce autologous skin left for wound repair. The purpose of this study was to explore how to effectively use the limited remaining autologous skin to repair the extensive deep wound.
Human keratinocytes harvested from the foreskin were cultured and transfected with epidermal growth factors (EGFs) by an adenovirus vector (Ad-EGF). The expression and the biological activity of EGF in both the normal human keratinocytes and the EGF gene-modified human keratinocytes were quantified by ELISA assay and CCK-8 assay, respectively. The differentiated phenotype of epidermal cells was detected by immunofluorescence staining via CK10, CK14, and CK19 expressions. Rats were subjected to a full-thickness skin loss (3.3 cm × 3.0 cm) on the dorsum, which was repaired with the EGF gene-modified human keratinocyte suspension and autologous microskin and covered with the allogeneic skin. The wound healing was quantified, and the expression of EGF mRNA was measured by RT-PCR.
The EGF gene-modified human keratinocytes highly expressed EGF. CK10, CK14, and CK19 as keratinocyte differentiation markers were increased in the EGF gene-modified human keratinocytes. Wound healing was accelerated remarkably by the combination of autologous microskin grafting and EGF gene-modified human keratinocytes in vivo, and a very high EGF mRNA expression was observed in EGF gene-modified human keratinocytes groups on days 7 and 14 compared with other groups.
DISCUSSION/CONCLUSION: The EGF gene-modified human keratinocyte suspension may serve as promising seed cells which can effectively secrete EGF to accelerate wound repair in combination with autologous microskin grafting and reduce the autologous skin requirement for wound repair.
大面积深度烧伤患者的高死亡率主要归因于广泛的烧伤创面以及用于创面修复的自体皮肤所剩无几。本研究的目的是探索如何有效利用有限的剩余自体皮肤来修复大面积深度创面。
从包皮获取人角质形成细胞，通过腺病毒载体（Ad-EGF）用表皮生长因子（EGF）进行转染。分别采用ELISA法和CCK-8法对正常人角质形成细胞和EGF基因修饰的人角质形成细胞中EGF的表达及生物学活性进行定量分析。通过CK10、CK14和CK19表达的免疫荧光染色检测表皮细胞的分化表型。对大鼠背部造成全层皮肤缺损（3.3 cm×3.0 cm），用EGF基因修饰的人角质形成细胞悬液和自体微粒皮进行修复，并用同种异体皮肤覆盖。对伤口愈合情况进行定量分析，通过RT-PCR检测EGF mRNA的表达。
EGF基因修饰的人角质形成细胞高表达EGF。作为角质形成细胞分化标志物的CK10、CK14和CK19在EGF基因修饰的人角质形成细胞中增加。在体内，自体微粒皮移植与EGF基因修饰的人角质形成细胞联合使用可显著加速伤口愈合，与其他组相比，在第7天和第14天，EGF基因修饰的人角质形成细胞组中观察到非常高的EGF mRNA表达。
讨论/结论：EGF基因修饰的人角质形成细胞悬液可能是有前景的种子细胞，其可有效分泌EGF，与自体微粒皮移植联合使用可加速伤口修复，并减少伤口修复所需的自体皮肤量。
