| Summary: | 碩士 === 高雄醫學大學 === 香粧品學系碩士班 === 104 === This manuscript demonstrated ex vivo and in vitro experimental effects on wound healing using three-dimensional porous scaffolds. Scaffold were constructed by natural and low-reject polymers, collagen (Col), hyaluronic acid (HA), and gelatin (Gel). The scaffolds were separated into two types according to HA concentrations, HA-L and HA-H. Our scaffold was cross-linked with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), and this scaffold was then tested by biocompatibility assessments, included ex vivo and in vitro characteristics. In chemical reaction, cross-linking groups occurred between the carboxyl groups of HA and the amino groups of Col (Gel), and Fourier transform infrared spectroscopy (FTIR) also detected the consistent results in raw materials and scaffolds. The swelling ratio of HA-L and HA-H scaffolds was analyzed by water absorption capability and displayed 31 and 36 scaffold weight enlargement. In biodegradation examination, scaffolds were degraded by three enzymes: lysozyme, hyaluronidase and collagenase I enzymes in time. The adipose-derived stem cell (ASCs) from rat adipocytes were detected the specific cluster-of-differentiation marker (positive: CD90, CD105 and CD146; negative: CD11b and CD31) by flow cytometry. ASCs were co-cultured within these scaffolds that promoted cell proliferation and stimulated capillary angiogenesis, such as transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF). Over the 7-day period, the cell growths of the scaffolds with HA-L and HA-H increased 3.5 and 4.2 folds more than on day 1. On day 7, the TGF-β concentrations of HA-L and HA-H were 36 and 39 pg/ml, and the VEGF concentrations were 12 and 57 pg/ml, respectively. The increase of VEGF concentration could be proven related to the scaffold HA amount. The morphology of ASCs in the scaffolds were inspected by fluorescence microscope and scanning electron microscope (SEM). In vivo histology examination, ASCs implanted scaffold transported into the subcutis, and the rat cells also proliferated into scaffolds. In histology section, the immune response rejection was no significant contrast between rat normal skin and ASCs scaffold. In the experimental data, the biocompatibility of HA-H scaffold was better than HA-L type, and therefore, ASCs HA-H scaffold improved wound repair efficiently.
Key words: wound healing; three-dimensional scaffolds; adipose-derived stem cells (ASCs); transforming growth factor-β (TGF-β); vascular endothelial growth factor (VEGF); in vivo
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