Department of Plastic Surgery, Affiliated Beijing Shijitan Hospital, Capital Medical University, Beijing, People's Republic of China; Department of Burn and Plastic Surgery, Burns Institute, The First Affiliated Hospital of PLA General Hospital, Beijing, People's Republic of China.
Department of Plastic Surgery, PLA General Hospital, Beijing, People's Republic of China.
Cytotherapy. 2014 Feb;16(2):160-9. doi: 10.1016/j.jcyt.2013.10.014.
Tissue-engineered dermis (TED) is thought to be the best treatment for skin defect wounds; however, lack of vascular structures in these products can cause slow vascularization or even transplant failure. We assessed the therapeutic potential of microencapsulated human umbilical cord mesenchymal stromal cells (hUCMSCs) expressing vascular endothelial growth factor (VEGF) in vascularization of TED.
hUCMSCs were isolated by means of enzymatic digestion and identified by means of testing biological characteristics. hUCMSCs were induced to differentiate into dermal fibroblasts in conditioned induction media. Collagen-chitosan laser drilling acellular dermal matrix (ADM) composite scaffold was prepared by means of the freeze dehydration and dehydrothermal cross-linking method. hUCMSC-derived fibroblasts were implanted on composite scaffolds to construct TED. TED with microencapsulated VEGF gene-modified hUCMSCs was then transplanted into skin defect wounds in pigs. The angiogenesis of TED at 1 week and status of wound healing at 3 weeks were observed.
The collagen-chitosan laser ADM composite has a uniform microporous structure. This composite has been used to grow hUCMSC-derived fibroblasts in vitro and to successfully construct stem cell-derived TED. Microencapsulated VEGF gene-modified hUCMSCs were prepared with the use of a sodium alginate-barium chloride one-step encapsulation technology. Seven days after the transplantation of the stem cell-derived TED and microencapsulated VEGF gene-modified hUCMSCs into the skin defect wounds on the backs of miniature pigs, the VEGF expression increased and the TED had a higher degree of vascularization. Re-epithelialization of the wound was completed after 3 weeks.
Microencapsulated VEGF gene-modified hUCMSCs can effectively improve the vascularization of TED and consequently the quality of wound healing.
组织工程真皮（TED）被认为是治疗皮肤缺损创面的最佳方法；然而，这些产品中缺乏血管结构会导致血管化缓慢，甚至移植失败。我们评估了表达血管内皮生长因子（VEGF）的微囊化人脐带间充质基质细胞（hUCMSCs）在 TED 血管化中的治疗潜力。
通过酶消化法分离 hUCMSCs，并通过生物学特性检测进行鉴定。hUCMSCs 在条件诱导培养基中诱导分化为真皮成纤维细胞。通过冷冻干燥和去水热交联法制备胶原-壳聚糖激光钻削脱细胞真皮基质（ADM）复合支架。将 hUCMSC 源性成纤维细胞植入复合支架上构建 TED。然后将微囊化 VEGF 基因修饰的 hUCMSCs 转染 TED 并移植到猪皮肤缺损创面。观察 1 周时 TED 的血管生成情况和 3 周时创面愈合情况。
胶原-壳聚糖激光 ADM 复合支架具有均匀的微孔结构。该复合支架已用于体外培养 hUCMSC 源性成纤维细胞，并成功构建干细胞源性 TED。采用海藻酸钠-氯化钡一步包埋技术制备微囊化 VEGF 基因修饰的 hUCMSCs。将干细胞源性 TED 和微囊化 VEGF 基因修饰的 hUCMSCs 移植到小型猪背部皮肤缺损创面 7 天后，VEGF 表达增加，TED 的血管化程度更高。3 周后，创面完成再上皮化。
微囊化 VEGF 基因修饰的 hUCMSCs 可有效改善 TED 的血管化，从而提高创面愈合质量。
