Department of Burns and Cutaneous Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China.
Burns. 2012 Aug;38(5):702-12. doi: 10.1016/j.burns.2011.12.014. Epub 2012 Feb 22.
This study aimed to investigate the feasibility of using an immortal keratinocyte cell line, HaCaT cells, to effectively deliver epidermal growth factor (EGF) in a skin substitute to treat burn wounds. The skin equivalent was constructed with human EGF (hEGF) gene modified HaCaT cells obtained through stable gene transfection; these were applied to full thickness burn wounds in a rat model. The results showed that the hEGF gene modified HaCaT cells produced more than 390ng/l of bioactive hEGF in the culture supernatant. K19 and integrin-β1 as keratinocyte differentiation markers were elevated in the hEGF gene modified HaCaT cells which were shown to be non-tumorigenic. The skin equivalent constructed with hEGF gene modified HaCaT cells demonstrated improved epidermal morphogenesis with a thick and compact epidermis. Wound healing was accelerated noticeably when applied with this skin substitute seeded with hEGF gene modified HaCaT cells in vivo. The results suggest that HaCaT cells modified with hEGF gene might be promising seed cells for construction of genetically modified skin substitute which can effectively secrete hEGF to accelerate wound repair and regeneration.
本研究旨在探讨利用永生化角质形成细胞系 HaCaT 细胞有效递送入皮生长因子（EGF）于皮肤替代物中以治疗烧伤创面的可行性。该表皮样组织是通过稳定的基因转染获得的人表皮生长因子（hEGF）基因修饰 HaCaT 细胞构建的；并将其应用于大鼠全层烧伤创面模型。结果表明，hEGF 基因修饰 HaCaT 细胞在培养上清液中产生超过 390ng/L 的生物活性 hEGF。hEGF 基因修饰 HaCaT 细胞中角蛋白细胞分化标志物 K19 和整合素-β1 升高，且显示非致瘤性。用 hEGF 基因修饰 HaCaT 细胞构建的表皮样组织显示出改善的表皮形态发生，具有厚而致密的表皮。体内应用接种 hEGF 基因修饰 HaCaT 细胞的皮肤替代物可明显加速创面愈合。结果表明，hEGF 基因修饰的 HaCaT 细胞可能是构建基因修饰皮肤替代物的有前途的种子细胞，可有效分泌 hEGF 以加速创面修复和再生。
