The Influence of bFGF on the Fabrication of Microencapsulated Cartilage Cells under Different Shaking Modes.

Xia Zhou,Ruimin Long,Yuangang Liu,Xiaolin Tang,Duanhua Cai,Shibin Wang,Pei Wang

doi:10.3390/polym11030471

Abstract

Cell encapsulation in hydrogels has been extensively used in cytotherapy, regenerative medicine, 3D cell culture, and tissue engineering. Herein, we fabricated microencapsulated cells through microcapsules loaded with C5.18 chondrocytes alginate/chitosan prepared by a high-voltage electrostatic method. Under optimized conditions, microencapsulated cells presented uniform size distribution, good sphericity, and a smooth surface with different cell densities. The particle size distribution was determined at 150–280 μm, with an average particle diameter of 220 μm. The microencapsulated cells were cultured under static, shaking, and 3D micro-gravity conditions with or without bFGF (basic fibroblast growth factor) treatment. The quantified detection (cell proliferation detection and glycosaminoglycan (GAG)/type II collagen (Col-II)) content was respectively determined by cell counting kit-8 assay (CCK-8) and dimethylmethylene blue (DMB)/Col-II secretion determination) and qualitative detection (acridine orange/ethidium bromide, hematoxylin-eosin, alcian blue, safranin-O, and immunohistochemistry staining) of these microencapsulated cells were evaluated. Results showed that microencapsulated C5.18 cells under three-dimensional microgravity conditions promoted cells to form large cell aggregates within 20 days by using bFGF, which provided the possibility for cartilage tissue constructs in vitro. It could be found from the cell viability (cell proliferation) and synthesis (content of GAG and Col-II) results that microencapsulated cells had a better cell proliferation under 3D micro-gravity conditions using bFGF than under 2D conditions (including static and shaking conditions). We anticipate that these results will be a benefit for the design and construction of cartilage regeneration in future tissue engineering applications.

Highlights

The concepts of “cytomedicine”, tissue engineering, and artificial organs have been developed in recent years, in which the heterotypic contact, cell-to-cell signaling, and uniform nutrient diffusion are key points
The results showed that cell encapsulation in hydrogels has been widely used in cytotherapy, regenerative medicine, 3D cell culture, and tissue engineering [6]
Masson stain kit was obtained from Jiancheng Technology Co., Ltd. (Nanjing, China) C5.18 cells, acridine orange/ethidium bromide (AO/EB) and klcian blue staining Kit were provided by Syagen Biosciences

Summary

Introduction

The concepts of “cytomedicine”, tissue engineering, and artificial organs have been developed in recent years, in which the heterotypic contact, cell-to-cell signaling, and uniform nutrient diffusion are key points. Artificial cells, called microencapsulated cells, can provide a liquid environment for cell cultures and maintain cell phenotype. They have potential for the immobilization of cells and enzymes, artificial organ translation, tissue engineering applications, etc. [1] As presented in 1933, Bisceglie et al sealed tumor cells in polymer membranes and transplanted them into the abdominal. The results showed that the cells could survive long enough without being damaged by the immune system. Lim et al [2] combined microencapsulation techniques with tissue-cell transplantation to prepare sodium alginate/poly-lysine/sodium alginate (APA) microcapsules with good biocompatibility as an immune isolation tool.

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Results

Conclusion