In vitro experimental research of basic fibroblast growth factor/chitosan/polylactic acid scaffolds in periodontal tissue regeneration
10.3969/j.issn.2095-4344.2017.26.002
- VernacularTitle:碱性成纤维细胞生长因子/壳聚糖/聚乳酸支架用于牙周组织再生工程的体外实验
- Author:
Lihong CHEN
;
Zhimin LEI
;
Lili WANG
- From:
Chinese Journal of Tissue Engineering Research
2017;21(26):4106-4112
- CountryChina
- Language:Chinese
-
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.
