含活性细胞的新型创面覆盖物的研制及初步应用  

Preparation and preliminary application of a new wound dressing containing living cells

作　　者：肖仕初[1] 杨珺[2] 贲道锋[1] 俞为荣[1] 王广庆[1] 杨勇[1] 朱世辉[1] 夏照帆[1] 

机构地区：[1]第二军医大学长海医院烧伤科,上海200433 [2]解放军第455医院整形科,上海200052

出　　处：《第二军医大学学报》2003年第10期1120-1123,共4页Academic Journal of Second Military Medical University

基　　金：国家杰出青年科学基金(39725029);上海市卫生系统百名跨世纪优秀学科带头人培养计划资助项目(97BR046).

摘　　要：目的:制备含活性细胞的高生物学活性的创面覆盖物,并观察其对创面愈合的促进作用。方法:提取鼠尾胶原,与硫酸软膏素按一定比例混合制成胶原膜;采用胶原酶消化培养法,取健康成人(15～35岁)环切包皮分离成纤维细胞培养并传代。将异体成纤维细胞种植于胶原膜表面培养1周,采用ELISA法和RIA法测定培养上清中细胞因子IL-6、IL-8、TGF-β1及细胞外基质层粘连蛋白、透明质酸的分泌;从需手术植皮的烧伤患者(18～45岁,烧伤面积5％～32％TBSA)大腿外侧切取厚约0.4mm的中厚皮片,将成纤维细胞-胶原膜覆盖物用于中厚皮片供皮区,观察愈合时间。结果:成纤维细胞种植于胶原膜表面生长良好,可长入膜内形成具有三维结构的覆盖物。培养1周,上清中测得IL-6为(502.2±21.9)pg/ml、IL-8为(340.5±23.2)pg/ml、TGF-β1为(1.431±0.063)pg/ml,层粘连蛋白为(92.1±13.5)ng/ml,透明质酸为(2 037.4±105.8)ng/ml。将覆盖物用于供皮区,其表皮化时间为(6.2±2.5)d,较单纯胶原膜的(9.8±2.3)d、凡士林纱布的(10.1±1.9)d明显缩短(P<0.01),上皮分化程度高。结论:合成纤维细胞的胶原膜具有较高的生物学活性,可作为一种良好的暂时性创面覆盖物。Objective:To prepare a highly bioactive wound dressing containing living cells and to observe their effects on wound healing. Methods: Allogeneic fibroblasts were seeded on the surface of collagen membrane and cultivated in vitro for 1 week. Secretion of interleukin-6 ,8(IL-6,8) and transforming growth factor-β1(TGF-β1) in the culture supernatants were assayed by enzyme linked immunosorbent assay. Larninin and hyaluronic acid in extracellular matrix were quantitated by radioactive immunoassay. Then, the wound dressing was used to cover donor sites for intermediate split thickness skin graft clinically; control donor sites received collagen membrane or vaseline gauze alone. Wound healing time were recorded, Results: Fibroblasts seeding on collagen membrane showed high proliferation, and grew into the space of membrane to form the three-dimensional structure. After 1 week, fibroblasts began to secrete IL-6(502. 2± 21. 9) pg/ml, IL-8 (340. 5±23. 2) pg/ ml, TGF-β1 (1. 431±0. 063) pg/ml, laminin (92. 1 ± 13. 5) ng/ml and hyaluronic acid (2 037. 4 ± 105. 8) ng/ml. The healing time of donor site covered with collagen membrane containing fibroblasts was shorter than those with collagen membrane and vaseline gauze[(6. 2 ± 2. 5) d vs (9. 8 ± 2. 3) and (10. 1 ± 1. 9) d], Conclusion: Collagen membrane seeded with fibroblasts shows good bioactivity and can serve as an excellent temporary wound dressing.

关 键 词：创面覆盖物 胶原膜 成纤维细胞 细胞因子 层粘连蛋白 透明质酸 创面愈合 

分 类 号：R641[医药卫生—外科学] R318.08[医药卫生—临床医学]