title: Decellularization of porcine skeletal muscle extracellular matrix for the formulation of a matrix hydrogel: a preliminary study.
abstract: Extracellular matrix (ECM) hydrogels are used as scaffolds to facilitate the repair and reconstruction of tissues. This study aimed to optimize the decellularization process of porcine skeletal muscle ECM and to formulate a matrix hydrogel scaffold. Five multi-step methods (methods A-E) were used to generate acellular ECM from porcine skeletal muscle [rinsing in SDS, trypsin, ethylenediaminetetraacetic acid (EDTA), Triton X-100 and/or sodium deoxycholate at 4-37°C]. The resulting ECM was evaluated using haematoxylin and eosin, 4-6-diamidino-2-phenylindole (DAPI) staining, and DNA quantification. Acellular matrix was dissolved in pepsin and gelled at 37°C. Hydrogel response to temperature was observed in vivo and in vitro. ECM components were assessed by Masson, Sirius red, and alcian blue staining, and total protein content. Acellular porcine skeletal muscle exhibited a uniform translucent white appearance. No intact nuclear residue was detected by haematoxylin and eosin staining, while DAPI staining showed a few nuclei in the matrixes produced by methods B, C, and D. Method A generated a gel that was too thin for gelation. However, the matrix obtained by rinsing in 0.2% trypsin/0.1% EDTA, 0.5% Triton X-100, and 1% Triton X-100/0.2% sodium deoxycholate was nuclei-free and produced a viscous solution that formed a structurally stable white jelly-like hydrogel. The residual DNA content of this solution was 49.37 ± 0.72 ng/mg, significantly less than in fresh skeletal muscle, and decreased to 19.22 ± 0.85 ng/mg after gelation (P < 0.05). The acellular matrix was rich in collagen and glycosaminoglycan, with a total protein concentration of 64.8 ± 6.9%. An acellular ECM hydrogel from porcine skeletal muscle was efficiently produced.
authors.affiliation: Department of Thyroid and Breast Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Yuehe
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Department of Thyroid and Breast Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Thyroid and Breast Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Chunxiang
authors.initials: C
authors.lastname: Tian
authors.affiliation: Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Jingcong
authors.initials: J
authors.lastname: Luo
authors.affiliation: Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Li
authors.initials: L
authors.lastname: Deng
authors.affiliation: Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Tingwu
authors.initials: T
authors.lastname: Qin
authors.affiliation: Department of Thyroid and Breast Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Lv
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000502:physiology; D003094:Collagen / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D003840:Deoxycholic Acid / Q000737:chemistry; D004492:Edetic Acid / Q000737:chemistry; D005109:Extracellular Matrix / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D007211:Indoles; D051379:Mice; D018482:Muscle, Skeletal / Q000737:chemistry*; D041681:NIH 3T3 Cells; D017830:Octoxynol / Q000737:chemistry; D044367:Phase Transition; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013552:Swine; D013696:Temperature; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014357:Trypsin / Q000737:chemistry
chemical_list: D006025:Glycosaminoglycans; D020100:Hydrogels; D007211:Indoles; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; C007293:DAPI; D017830:Octoxynol; D003094:Collagen; D004247:DNA; D004492:Edetic Acid; D014357:Trypsin
keywords: extracellular matrix; hydrogel; skeletal muscle; tissue engineering; tissue scaffold