基于脂肪干细胞的皮肤皮脂腺三维结构的重建及机制研究

Reconstruction of sebaceous glands is one of the goals to achieve functional healing on patients suffered severe burns instead of the simple pursuit of closure of the wound. The exploratory study on the formation of its three-dimensional structure in vitro will lay the foundation for the reconstruction of sebaceous glands. Studies have shown that human adipose tissue-derived stem cells(ADSCs) which are easy to be drawn and have broad source, possess powerful plasticity of across-germinal layer development and multi-directional differentiation(include adipogenic cells).Activation of C-myc and β-catenin plays an important role in sebaceous differentiation. Based on our early work that hepatocyte growth factor (HGF) and androgen(5ɑ-DHT) can induce sebaceous gland tube, we will firstly construct HGF and androgen microcapsules using micro capsule slow-release technology, and induct human ADSCs successively to observe human ADSCs migration, morphology and protein expression of NF-kappa B, ERK and AKT signaling pathways which are related to canaliculization after induced. On the basis of promoting the expression of c-myc and β-catenin, We will observe the three-dimensional structure formation of human ADSCs after induction with microcarrier culture technology. Accordingly we will construct the tissue-engineering skin with regeneration potentiality of three-dimensional structure of cutaneous sebaceous gland, in order to provide a basis for achieving functional tissue-engineered skin.

重建皮脂腺是严重烧伤后由单纯追求封闭创面到实现功能性愈合的目标之一，对皮脂腺体外三维结构的重建研究可为皮脂腺的重建奠定基础。研究表明易于取材、来源广泛的脂肪干细胞（ADSCs）具有强大的跨胚层发育可塑性及多向（包括成脂）分化能力；同时c-myc和β-catenin的活化在皮脂腺分化中起重要作用。我们在前期应用肝细胞生长因子（HGF）和雄激素（5ɑ-双氢睾酮）促进体外诱导皮脂腺腺管发生的基础上，拟构建HGF和雄激素缓释微囊并持续诱导分化人ADSCs，观察其移行、传代和形态的变化，并检测与小管形成相关的NF-κB、ERK、AKT等信号通路蛋白的表达情况；在此基础上促进c-myc和β-catenin表达，利用微载体培养技术观察经过上述诱导分化后ADSCs形成三维结构情况，并根据前述实验构建具有皮脂腺三维结构再生潜能的组织工程皮肤，从而为体外重建皮脂腺和实现功能化组织工程皮肤奠定基础。

重建皮脂腺是严重烧伤后由单纯追求封闭创面到实现功能性愈合的目标之一，对皮脂腺体外三维结构的重建研究可为皮脂腺的重建奠定基础。本研究中我们制备HGF和5ɑ-双氢睾酮缓释胶囊，作用于具有多种分化潜能的ADSCS细胞，通过检测处理后的ADSCS细胞的增殖能力、迁移能力、形态变化以及与成管有关的NF-κB、ERK、AKT信号通路，确定了HGF和5ɑ-双氢睾酮体外诱导皮脂腺腺管发生的作用；在此基础上，我们加入促进c-myc表达的AVE0991和促进β-catenin表达的曲格列酮，使得ADSCS细胞向上皮细胞转化。经过诱导的ADSCS细胞在羊膜微粒上培养后孵育在创面上，能有效的促进创面的愈合。