Decellularized porcine skin matrix for tissue-engineered meniscus scaffold

doi:10.3969/j.issn.2095-4344.3173

Abstract

Abstract: BACKGROUND: Regeneration and repair of the injured meniscus remain scientific challenges. It is of significance to construct a tissue-engineered scaffold with good biomechanical performance and cytocompatibility.
OBJECTIVE: To prepare decellularized porcine skin matrix scaffold and evaluate its biomechanical properties and cytocompatibility.
METHODS: Porcine skin was decellularized with the combination of trypsin, Triton X-100 and sodium dodecyl sulfate to obtain decellularized porcine skin matrix. DAPI staining was used to observe whether the cells were well removed and scanning electron microscope to observe its microstructure. The decellularized porcine skin matrix was compared with natural meniscus in term of histological staining, contents of collagen and glycosaminoglycans, and biomechanical properties. The third-generation bone marrow mesenchymal stem cells were seeded on the decellularized porcine skin matrix. After 3 days of culture, the scanning electron microscopy, dead/live cell staining and phalloidin staining were used to observe cytocompatibility of the decellularized porcine skin matrix.
RESULTS AND CONCLUSION: (1) DAPI staining showed that no cell nucleus was found in the decellularized porcine skin matrix. Scanning electron microscopy showed that the surface of the decellularized porcine skin matrix was rough, and the surface fibers showed a topological structure. (2) Histological staining showed that the natural meniscus tissue was rich in extracellular matrix, and the meniscus cells were evenly distributed in the matrix. The collagen fibers were dense and arranged regularly, and the collagen composition was mainly type I collagen fibers and contained glycosaminoglycan components. The decellularized porcine skin matrix had no cell nucleus and was in the shape of loose fiber strips with more pore structure, and its collagen component was mainly type I collagen fiber and contained glycosaminoglycan components. (3) There was no significant difference in collagen content and tensile modulus between decellularized porcine skin matrix and natural meniscus (P > 0.05), while the glycosaminoglycan content and compression modulus were lower than those of the natural meniscus (P < 0.05). The biomechanical properties of the decellularized porcine skin matrix are sufficient to bear the load of the knee joint and maintain the integrity of the scaffold. (4) Scanning electron microscope showed that bone marrow mesenchymal stem cells adhered to the pore structure of porcine skin matrix, and the cells were elliptical and grew in clusters. (5) Dead/live cell staining showed that the bone marrow mesenchymal stem cells on the decellularized porcine skin matrix had high activity (94.0±3.8)%. (6) Phalloidin/DAPI staining showed that bone marrow mesenchymal stem cells showed good spreading on the decellularized porcine skin matrix. (7) The results showed that decellularized porcine skin matrix has similar collagen components as the natural meniscus, and presents good biomechanical performance and cytocompatibility, and is conducive to cell adhesion and growth.

Key words: meniscus, tissue engineering, decellularized scaffold, porcine skin matrix, collagen, extracellular matrix

CLC Number:

Wang Hao, Chen Mingxue, Li Junkang, Luo Xujiang, Peng Liqing, Li Huo, Huang Bo, Tian Guangzhao, Liu Shuyun, Sui Xiang, Huang Jingxiang, Guo Quanyi, Lu Xiaobo. Decellularized porcine skin matrix for tissue-engineered meniscus scaffold[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(22): 3473-3478.

Figures/Tables 9

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