Preparation of platelet-derived growth factor loaded three-dimensional bio-printed Meniscus scaffold

doi:10.12307/2021.059

Abstract

Abstract: BACKGROUND: Based on the need of in situ tissue engineering regeneration in the field of meniscus regeneration, three-dimensional (3D) bioprinting technology can be used to construct a scaffold which can effectively combine recruitment bioactive factors and excellent materials.
OBJECTIVE: To optimize the preparation process of polycaprolactone/methacrylate anhydride gelatin/meniscus extracellular matrix (PCL/GelMA/MECM) 3D bioprinting scaffold loaded with platelets derived growth factor-BB (PDGF-BB), and to test the sustained release properties of the growth factor and its effect on the migration and proliferation of the synovial mesenchymal stem cells.
METHODS: Photosensitive bio-ink was prepared by GelMA/MECM. PCL scaffold and PCL/GelMA/MECM bio-scaffold were prepared by 3D bioprinting technology. Finally, PDGF-BB was loaded to fabricate functional bioprinting PCL/GelMA/MECM/PDGF scaffold, and then its mechanical properties and sustained release properties were tested. New Zealand white rabbit synovial mesenchymal stem cells were isolated and cultured. Transwell and CCK-8 assays were used to explore the effects of PCL scaffold, PCL/GelMA/MECM scaffold and PCL/GelMA/MECM/PDGF scaffold on the migration and proliferation of synovial mesenchymal stem cells, and single medium cultured cells were used as negative control. The third passage of synovial mesenchymal stem cells was inoculated on PCL scaffold, PCL/GelMA/MECM scaffold and PCL/GelMA/MECM/PDGF scaffold respectively. The viability, adhesion and growth of the cells on the three scaffolds were observed by confocal microscope and scanning electron microscope.
RESULTS AND CONCLUSION: (1) There was no significant difference in tensile modulus and compression modulus between PCL/GelMA/MECM scaffolds and PCL scaffolds (P > 0.05). (2) PCL/GelMA/MECM/PDGF scaffolds had good sustained release performance, and PDGF-BB could be released continuously for up to 28 days. (3) Compared with the negative control group, PCL/GelMA/MECM scaffolds and PCL/GelMA/MECM/PDGF scaffolds could promote the migration of synovial mesenchymal stem cells, among which PCL/GelMA/MECM/PDGF scaffolds were more effective. (4) Compared with the negative control group, the cells cultured with PCL/GelMA/MECM scaffolds and PCL/GelMA/MECM/PDGF scaffolds proliferated faster after 4 and 7 days (P < 0.001), and the proliferative effect of PCL/GelMA/MECM/PDGF scaffolds was more obvious. (5) Dead/Live staining showed that the cells in each group had good activity, and the number of cells on the PCL/GelMA/MECM/PDGF scaffold was the highest. (6) Scanning electron microscope showed that the cells were attached to the surface of the three groups of scaffolds, and the cells on PCL/GelMA/MECM scaffolds and PCL/GelMA/MECM/PDGF scaffolds adhered to a large number of cells and secreted a large amount of extracellular matrix. (7) These results showed that the 3D bioprinted meniscus scaffold loaded with PDGF-BB had good mechanical properties and biocompatibility and could promote the migration and proliferation of synovial mesenchymal stem cells.

Key words: bone, materials, meniscus, extracellular matrix, platelet-derived growth factor, 3D bioprinting, synovial mesenchymal stem cells, migration, proliferation

CLC Number:

李浩, 杨振, 高仓健, 付力伟, 苑志国, 眭翔, 刘舒云, 郭全义. Preparation of platelet-derived growth factor loaded three-dimensional bio-printed Meniscus scaffold[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(28): 4465-4472.

Figures/Tables 8

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