组织工程微粒真皮的构建及其动物实验研究

Construction and in vivo study of tissue engineering granule dermis

目的:将人成纤维细胞和York猪脱细胞真皮基质(ADM)微粒复合构建的?组织工程微粒真皮应用于SD大鼠损伤模型,观察其修复组织创伤能力。方法:用PKH26标记的人成纤维细胞,与猪ADM微粒复合构建微粒真皮,将其注射到SD大鼠皮下,用荧光显微镜和HE观察ADM微粒真皮对皮下组织的修复;将54只SD大鼠随机分成3组,分别移植自体全厚皮肤、ADM微粒和ADM微粒皮真皮。大体、荧光显微镜和HE观察伤口愈合情况、计算创面愈合时间和收缩率。结果:皮下注射ADM微粒真皮2周时可见单核细胞、成纤维细胞和血管长入,以后单核细胞减少,荧光观察可见红色荧光,8周时注射的微粒真皮基本被自身组织改建利用、体积稳定,胶原粗大整齐;ADM微粒真皮移植能够使创面愈合时间、创面收缩率与自体全厚皮肤移植相比无统计学意义(P>0.05),与ADM微粒移植相比差异显著(P<0.01)。结论:皮下注射ADM微粒真皮可参与皮下组织改建,修补皮下组织缺损;ADM微粒真皮可促进SD大鼠损伤模型的愈合和瘢痕挛缩。

Objective To explore the in vivo curative effect of tissue engineering granule dermis constructed with human fibroblasts and acellular dermal matrix （ADM） granules via subcutaneous injection and full-thickness skin wound transplantation. Methods Human fibroblasts were cultured in vitro and labeled with PKH26, then combined with ADM granules to construct tissue engineering granule dermis. Restoration of subcutaneous tissue after tissue engineering granule dermis injection was measured via fluorescent microscope and HE histological assay; simultaneity restoration of skin on SD rat full-thickness skin wound models after autograft, ADM and tissue engineering granule dermis transplantation were measured via macroscopy, fluorescent microscope and HE histological assay. Results Fibroblasts and ADM granules were combined well： cells were proliferated, layered-adhered and ECM deposited well on ADM.after 2-week subcutaneous injection,cells and vascular tissue were ingrown and inflammatory cells were lessen with time, red fluorescence was observed, 8 weeks later, injected tissues were rebuilt and collagen was regular.There is no statistical differences （P 〉 0.05） between autograft and tissue engineering granule dermis transplantation on healing time and wound contraction, while tissue engineering granule dermis is better than ADM alone （P 〈 0.01）. Conclusion The efficient tissue engineering granule dermis could be constructed with human fibroblasts and ADM granules well. Aceliular dermal matrix （ADM） granules dermis could improve in vivo restoration of subcutaneous tissue and skin wound via injection and full-thickness skin wound transplantation.