Microencapsulated VEGF gene-modified umbilical cord mesenchymal stromal cells promote the vascularization of tissue-engineered dermis: an experimental study
Microencapsulated VEGF gene-modified umbilical cord mesenchymal stromal cells promote the vascularization of tissue-engineered dermis: an experimental study
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DOI：
10.1016/j.jcyt.2013.10.014
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发表时间：
2014-02-01
期刊：
影响因子：
4.5
通讯作者：
Liu, Jing
中科院分区：
文献类型：
--
作者：
Han, Yanfu;Tao, Ran;Liu, Jing
背景目标。组织工程真皮（TED）被认为是皮肤缺损伤口的最佳治疗方法;然而，这些产品中缺乏血管结构可能导致血管形成缓慢甚至移植失败。我们评估了微囊化表达血管内皮生长因子（VEGF）的人脐带间充质基质细胞（hUCMSCs）在TED血管化中的治疗潜力。方法.酶消化法分离hUCMSCs，生物学特性检测法鉴定hUCMSCs。在条件诱导培养基中诱导hUCMSC分化为真皮成纤维细胞。采用冷冻脱水-脱水交联法制备胶原-壳聚糖激光打孔脱细胞真皮基质（ADM）复合支架。将hUCMSC衍生的成纤维细胞植入复合支架上以构建TED。然后将微囊化VEGF基因修饰的hUCMSCs的TED移植到猪的皮肤缺损伤口中。观察1周时TED血管生成情况和3周时创面愈合情况。结果胶原-壳聚糖激光ADM复合物具有均匀的微孔结构。该复合物已用于体外培养hUCMSC衍生的成纤维细胞，并成功构建干细胞衍生的TED。采用海藻酸钠-氯化钡一步包埋法制备微囊化VEGF基因修饰的hUCMSCs。将干细胞来源的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后7天，VEGF表达增加，TED具有更高程度的血管化。3周后完成伤口的再上皮化。结论.微囊化VEGF基因修饰的hUCMSCs能有效促进TED血管化，从而提高创面愈合质量。
Background aims. 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. Methods. 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. Results. 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. Conclusions. Microencapsulated VEGF gene-modified hUCMSCs can effectively improve the vascularization of TED and consequently the quality of wound healing.