Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair

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 scaffold...

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Main Authors: Hung-Hsun Yen, 顏宏勳
Other Authors: Hui-Min David Wang
Format: Others
Language:zh-TW
Published: 2016
Online Access:http://ndltd.ncl.edu.tw/handle/73548157085385954795
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spelling ndltd-TW-104KMC057930012017-09-24T04:40:52Z http://ndltd.ncl.edu.tw/handle/73548157085385954795 Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair 以脂源性幹細胞培養在具新穎性生物相容性孔洞型支架促進傷口修復 Hung-Hsun Yen 顏宏勳 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 Hui-Min David Wang 王惠民 2016 學位論文 ; thesis 49 zh-TW
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language zh-TW
format Others
sources NDLTD
description 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
author2 Hui-Min David Wang
author_facet Hui-Min David Wang
Hung-Hsun Yen
顏宏勳
author Hung-Hsun Yen
顏宏勳
spellingShingle Hung-Hsun Yen
顏宏勳
Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
author_sort Hung-Hsun Yen
title Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
title_short Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
title_full Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
title_fullStr Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
title_full_unstemmed Adipose-Derived Stem Cells Cultured in Novel Biocompatible Porous Scaffold for Wound Repair
title_sort adipose-derived stem cells cultured in novel biocompatible porous scaffold for wound repair
publishDate 2016
url http://ndltd.ncl.edu.tw/handle/73548157085385954795
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