In vitro experimental research of basic fibroblast growth factor/chitosan/polylactic acid scaffolds in periodontal tissue regeneration

doi:10.3969/j.issn.2095-4344.2017.26.002

Abstract

Abstract:

BACKGROUND: Basic fibroblast growth factor (bFGF) can promote mitosis and chemotaxis of periodontal ligament fibroblasts (PDLCs), and help the generation of extracellular matrix and new blood capillaries. But it is easy to degrade, has short half-life, and metabolizes rapidly.

OBJECTIVE: To evaluate the influence of bFGF/chitosan/polylactic acid scaffolds on PDLCs growth and proliferation.

METHODS: Polylactic acid/chitosan composite scaffolds in different proportions (4:1, 3:2, 1:1, 2:3, 1:4) were prepared through freeze-drying method, to study their microstructure, porosity and mechanical strength and then choose the optimal ratio of chitosan/polylactic acid scaffold that was used to prepare the composite scaffolds which contained different concentrations (0, 0.1, 1, 10, 100 µg/L) of bFGF. (1) Cytotoxicity test: PDLCs were cultured with leaching liquid of polylactic acid/chitosan scaffolds which contained different concentrations (0, 0.1, 1, 10, 100 µg/L) of bFGF and DMEM culture medium respectively. The effects on cell proliferation were tested by MTT after 24, 48, 72 hours. Cell cycles were tested using flow cytometry at 5 days of culture. (2) Cytocompatibility test: PDLCs were co-cultured with the polylactic acid/chitosan scaffolds which contained different concentrations (0, 0.1, 1, 10, 100 µg/L) of bFGF. The number of PDLCs was counted at 1, 3, 5 and 7 days. The growing status of PDLCs on the scaffolds was observed by scanning electron microscope at 3 days of culture.

RESULTS AND CONCLUSION: (1) The best mass ratio of polylactic acid and chitosan was 2:3 by test of porosity and mechanical strength. (2) The absorbance value of each group was increased over time. The absorbance values of 0.1, 1, 10, 100 µg/L bFGF groups were higher than that of the control group (0 µg/L bGFG), and the A value of 10 µg/L group was highest in all groups at 48 and 72 hours after co-culture (P < 0.05). (3) Cell percentages at G₀/G₁ phase of 0.1, 1, 10, 100 µg/L bFGF groups were higher than that of the control group, and the percentage of 10 µg/L group was lowest in all groups (P < 0.05). Cell percentage at S phase and G₂/M+S of 0.1, 1, 10, 100 µg/L bFGF groups were higher than that of the control group, and the percentage of 10 µg/L group was highest in all groups (P < 0.05). (4) The number of cells in each group was increased with time. The cell number in the 10 µg/L was most in all groups at 3 and 5 days of co-culture (P < 0.05). Observation of scanning electron microscopy showed that PDLCs adhered and grew well on the 10 µg/L bFGF/polylactic acid/chitosan composite scaffold when co-cultured for 3 days. Overall, these findings indicate that the bFGF/polylactic acid/chitosan composite scaffold contributes to PDLCs proliferation.

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

Key words: Periodontium, Chitosan, Fibroblast Growth Factor 2

CLC Number:

R318

Chen Li-hong, Lei Zhi-min, Wang Li-li.

In vitro experimental research of basic fibroblast growth factor/chitosan/polylactic acid scaffolds in periodontal tissue regeneration

[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(26): 4106-4112.

Figures/Tables 10

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