Release and effect of growth factors conjugated into collagen patches on human bone marrow mesenchymal stem cell proliferation and differentiation

doi:10.3969/j.issn.2095-4344.2014.52.020

Abstract

Abstract:

BACKGROUND: Preclinical trials in animals have demonstrated myocardial cell transplantation can improve heart function, but it is hindered by the low retention of transplanted cells. Therefore, it becomes a common strategy to generate a proper cell platform to promote cell retention and vasculature formation.

OBJECTIVE: To evaluate the cytokine release of a novel cytokine-conjugated collagen patch prepared with 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) and its physiological effects on proliferation and differentiation of seeded human bone marrow mesenchymal stem cells.

METHODS: After activating and crosslinking by EDC, the control patches (EDC-patch) were preserved in PBS and the experimental patches (GF-patch) were immersed into PBS containing vascular endothelial growth factor and basic fibroblast growth factor for cytokine conjugation. Collagen patches without EDC treatment underwent same process as internal control. ELISA was performed to detect the content of vascular endothelial growth factor and basic fibroblast growth factor in the supernatant of GF-patch at 1, 3, 7 days, 2, 3, 4 weeks after patch preparation. The 0.5×10⁶ human bone marrow mesenchymal stem cells were evenly seeded on both kinds of patches and the cell proliferation was identified by hematoxylin-eosin staining followed by cell counting, MTT assay or BrdU staining test. Accordingly, cell differentiation was illustrated by RT-PCR for collagen I and collagen III expression, and immunofluorescent staining for SMA in seeding cells.

RESULTS AND CONCLUSION: 42.4% vascular endothelial growth factors and 24.5% basic fibroblast growth factors were successfully conjugated into collagen patches and they both exhibited the constant, controlled-release mode during the 4-week observation period. By contrast, the patch without EDC treatment just showed the physical bonding of both cytokines. In comparison with EDC-patch, GF-patch presented the capability to induce cell proliferation while retard cell differentiation.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: collagen, carbodiimides, vascular endothelial growth factors, fibroblast growth factor 2

CLC Number:

R318

Chuai Jun-bo, Kang Kai, Sun Lu, Qu Hui, Cai Jun, Chen Ke-gong, Xie Bao-dong, Jiang Shu-lin, Tian Hai, Li Ren-ke. Release and effect of growth factors conjugated into collagen patches on human bone marrow mesenchymal stem cell proliferation and differentiation [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(52): 8474-8479.

Figures/Tables 2

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