组织工程化表皮膜片构建及其在供皮区的应用研究

Construction and clinical application of tissue engineered epidermal membrane

目的构建组织工程化表皮细胞膜片并应用于供皮区创面治疗。方法利用患者少量自体或同种异体正常皮肤分离、培养、扩增的表皮细胞,接种于壳聚糖明胶膜构建成表皮细胞膜片;将膜片移植于治疗组供皮区创面、适度加压,同时设立对照组:空白材料对照以及传统油纱布对照覆盖创面。于术后5～10d、30d、90d行大体观察、组织学检查、广谱角蛋白、外皮蛋白、层粘连蛋白、免疫组织化学检测以及Ⅰ、Ⅲ型胶原比值测定(偏光显微镜、RTPCR)。结果利用自体及异体表皮细胞和壳聚糖明胶膜能够构建出人表皮细胞膜片,应用于临床供皮区创面治疗15例,经过3个月随访,疗效肯定。移植膜片创面愈合时间(8.1±1.3)d,空白材料对照区为(16.2±3.8)d,空白油纱布对照区为(23.0±5.8)d。移植膜片存活良好,结构较完整、术后30d及90d观察:12例无明显增生,3例有轻度增生(20.0%),而空白油纱布对照区11例遗留增生性瘢痕(74.4%,χ2=8.127,P<0.01)。结论表皮细胞-壳聚糖明胶膜片能促进供皮区创面早期愈合并减少后期瘢痕增生,具有良好治疗效果。

Objective To Construct tissue engineered epidermal membrane for promoting healing of clinical skin graft donor site wound. Methods Epidermal cells /Chitosan-Gelatin (CG)membrane was constructed with cultured human epidermal cells(EC) and CG membrane, at the donor site of split skin graft, the wound was divided into three groups: the control group was covered with CG membrane without KC , KC/CG membrane was grafted on the treatment part of the wound area , and blank group was covered with traditional vaseline gauze.The engineered epidermal membrane and its effect on wound were evaluated with gross observation, HE , immunohistochemistry , collagen type Ⅰ/Ⅲ ratio by picrosirius polarization method and RT-PCR test at various time points before and after operation. Results The result showed that human tissue engineered epidermis could be constructed with cultured human EC and CG membrane ,and this artificial epidermal membrane could be used for promoting the healing of skin graft donor site wound successfuly (16 cases with 3 monthes'oberservation). The average healing time is (16.2±3.8)days for control group,(8.1±1.3)days for experimental group and (23.0±5.7)days for blank group. The artificial epidermis was well survived with normal structure. In addition, less hypertrophic scar develement was observed in treated wound at 90 days (3 in 16 cases,20.0%) than in the blank sites (11 in 16 cases, 74.4%). The difference is statistically significant (χ~2=8.127,P<0.01). Conclusions The constructed EC-CG membrane can be constructed in vitro,survived in vivo and has good clinical application in promoting healing of skin graft donor site wound and inhibiting hypertrophic scar formation.