Enhanced decellularization technique of porcine dermal ECM for tissue engineering applications
- Authors
- Ventura, Reiza D.; Padalhin, Andrew R.; Park, Chan Mi; Lee, Byong Taek
- Issue Date
- Nov-2019
- Publisher
- Elsevier BV
- Keywords
- Extracellular matrix; Porcine skin; Isopropanol; Decellularization
- Citation
- Materials Science and Engineering: C, v.104
- Journal Title
- Materials Science and Engineering: C
- Volume
- 104
- URI
- https://scholarworks.bwise.kr/sch/handle/2021.sw.sch/4132
- DOI
- 10.1016/j.msec.2019.109841
- ISSN
- 0928-4931
1873-0191
- Abstract
- Effective removal of cellular components while retaining extracellular matrix (ECM) proteins is the ultimate goal of decellularization. The aim of this study is to produce a decellularized ECM with highly preserved ECM proteins and to determine the effect of isopropanol as a decellularization solvent on the characteristics of the decellularized porcine skin. Two different protocols were used for porcine skin decellularization. Protocol 1 consisted of Triton-X and sodium dodecyl sulfate (SDS) in water while protocol 2 consisted of Triton-X and SDS in 70% isopropanol. After decellularization, DNA components decreased significantly in protocol 2 with lower amount of lipid content and higher ECM proteins such as collagen (92.91 +/- 9.02 mu g/mg sample), alpha-elastin (142.32 +/- 6.74 mu g/mg sample) and sulfated glycosaminoglycan (sGAG; 7.44 +/- 1.30 mu g/mg sample) compared with protocol 1 ECM. Higher amount of vascular endothelial growth factor (VEGF; 11.26 +/- 0.44 pg/mg sample) content was quantified in protocol 2 compared with protocol 1 while higher trace amount of bone morphogenic protein 2 (BMP-2; 0.28 +/- 0.04 pg/mg sample) was also observed in protocol 2 compared with protocol 1. Protocol 2 ECM did not significantly affect the cell viability and exhibited no cytotoxicity when exposed to three different cell lines: L929 fibroblast cells, MC3T3-E1 pre-osteoblast cells, and rat mesenchymal stem cells (BMSC). Subcutaneous implantation after 7 and 21 days revealed higher cell infiltration in protocol 2 ECM and enhanced neovascularization. Isopropanol/surfactants proved to be effective in cell and lipid removal during decellularization while preserving the higher amount of ECM proteins.
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Collections - College of Medicine > Department of Regenerative Medicine > 1. Journal Articles
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