Extracellular matrix from human fetal progenitor cells improves wound healing
- 주제(키워드) Fetal cartilage derived progenitor cells , Conditioned medium , Extracellular matrix , 3D culture , Dialysis membrane , Lyophilization , Wound healing
- 발행기관 아주대학교
- 지도교수 Byoung-Hyun Min
- 발행년도 2020
- 학위수여년월 2020. 8
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000030192
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Burn injury is the main cause more than 70% scar formation for patients. In previous study has been reported fetal stem cells conditioned medium significantly promoted skin regeneration compared to mesenchymal stromal cells (MSCs). However, little is known about the functionality of key factors and mechanisms through the analysis of conditioned medium from fetal stem cells. In this study, we aimed to fabricate a biologically functional human fetal cartilage derived progenitor cells-conditioned medium (hFCPCs-CM) that can improve wound healing and minimize scar formation on a burn skin injury model in rats. The hFCPCs-CM was prepared by a series of processes including a 3D culture of hFCPCs, a 3.5 kDa cutoff dialysis membrane of conditioned medium and lyophilization of extracellular matrix. The final lyophilized hFCPCs-CM did not contain the cellular content and contained large amount of total protein, collagen, glycosaminoglycans and soluble proteins of IGFBP-2, IGFBP-6, HGF, VEGF, TGF β3 and M-CSF compared to human bone marrow-derived MSCs-conditioned medium (hBMSCs-CM). It also contained 2 specific proteins of collagen alpha-1(XIV) chain and collagen alpha-2(I) chain. The acceleration effect of hFCPCs-CM on wound healing was more prominent than hBMSCs-CM, in enhancing the viability, proliferation and migration of fibroblast and polarization of M2 macrophages in vitro. The therapeutic potential of the hFCPCs-CM was also observed better than that of hBMSCs-CM in a burn skin injury model in rats by efficiently accelerating wound healing and minimize scar formation. Our results show that the hFCPCs-CM could be a potential therapeutic approach to promote wound healing process.
more목차
1. Introduction 1
2. Materials and Methods 6
2.1 Preparation of the hFCPCs-CM 6
2.2 Biochemical Characterization of the hFCPCs-CM 8
2.3 Growth factor detection and evaluation in the hFCPCs-CM 10
2.4 Effects of hFCPCs-CM on fibroblast viability, proliferation and migration 10
2.5 Effects of hFCPCs-CM on human macrophage polarization 12
2.6 Effects of hFCPCs-CM on skin burn wound healing using a rat model 14
2.7 Statistical analysis 16
3. Results 18
3.1 Preparation of hFCPCs-CM 18
3.2 Biochemical characterization of hFCPCs-CM 20
3.3 The unique proteins in conditioned medium produced by hFCPCs 22
3.4 Detection of secreted growth factors in hFCPCs-CM 24
3.5 Effects of hFCPCs-CM on fibroblast viability, proliferation and migration 26
3.6 Effects of hFCPCs-CM on macrophage polarization in vitro 29
3.7 Established rat skin burn wound model, wound closure rate and scar formation 32
3.8 Effects of hFCPCs-CM on epidermal thickness index and scar elevation index changes 37
3.9 Effects of hFCPCs-CM on collagen deposition, organization, and density of collagen type I changes 40
3.10 Effects of hFCPCs-CM on macrophage polarization in vivo 43
4. Discussion 45
5. Conclusion 49
References 50
