Effects of oxygen concentration on proliferation, apoptosis, and migration ability of in vitro cultured rabbit auricle chondrocytes
10.3760/cma.j.cn114453-20210417-00168
- VernacularTitle:不同氧浓度对兔耳软骨细胞增殖、凋亡、迁移功能的影响
- Author:
Chen YANG
1
;
Jinxiu YANG
;
Haiyue JIANG
;
Leren HE
Author Information
1. 中国医学科学院北京协和医学院整形外科医院耳再造一中心 100144
- Keywords:
Chondrocyte;
Tissue engineering;
Cell proliferation;
Apoptosis;
Cell movement
- From:
Chinese Journal of Plastic Surgery
2021;37(7):802-809
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of oxygen concentration on proliferation, apoptosis and migration of rabbit auricular chondrocytes at different time points of culture.Methods:Bilateral auricular cartilage from 6 Japanese white rabbits were harvested and cultured, auricular chondrocytes at passage 2 (P2) were used in this study. Five groups were set: Group A (control group): chondrocytes were cultured in atmospheric (21%) oxygen condition, group B: chondrocytes were cultured in 5% oxygen condition for 12 hours (h), group C: chondrocytes were cultured in 5% oxygen condition for 36 h, group D: chondrocytes were cultured in 1% oxygen condition for 12 h, and group E: chondrocytes were cultured in 1% oxygen condition for 36 h. Cell counting kit-8 (CCK-8) assay was adopted to evaluate the proliferation of chondrocytes. Proliferation curves were drawn according to the absorbance value detected. Cell apoptosis was investigated by annexin V-FITC and flow cytometry. Cell scratch test was used to observe the migration ability. One-way ANOVA and LSD- t were used to verify differences between the groups. P<0.05 was considered as statistically significant. Results:In terms of cell proliferation, on the sixth day of culture, the absorbance values of group A, B, C, D and E were 1.38 ± 0.29, 1.59 ± 0.27, 1.37 ± 0.22, 2.06 ± 0.64, 2.23 ± 0.56 respectively. Significant difference was found between the five groups ( F=4.207, P=0.012), and there were significant differences between group D and A ( t=-2.487, P=0.022), group E and A ( t=-3.095, P=0.006). On the seventh day of culture, the absorbance values of the five groups were 1.72 ± 0.30, 2.26 ± 0.44, 2.30 ± 0.29, 2.49 ± 0.73, 2.74 ± 0.54. Significant difference was found between five groups ( F=2.948, P=0.046), and there were significant differences between group D and A ( t=-2.480, P=0.022), group E and A ( t=-3.287, P=0.004). The apoptosis rate of group A, B, C, D and E were (2.97±1.14)%, (3.92±1.14)%, (3.38±0.83)%, (1.54±0.50)%, (0.99±0.59)%. The difference between groups for cell apoptosis were also significant ( F=7.957, P=0.001), among which there were significant differences between group D and A ( t=-2.300, P=0.036), group E and A ( t=-3.180, P=0.006), group B and D ( t=3.812, P=0.002), group C and E ( t=3.832, P=0.002). As for cell migration, at the 12th hour, the cell migration rates of groups A, B, C, D and E were(13.10±3.32)%, (9.23±3.56)%, (10.60±2.03)%, (13.33±1.72)%, (15.32±4.72)%. Significant difference was found between the five groups ( F=3.278, P=0.027). And there were significant differences between group D and B ( t=2.183, P=0.039), group C and E ( t=-2.513, P=0.019). At 24 h, the cell migration rates of the five groups were(19.7±2.97)%, (16.62±3.30)%, (18.99±2.61)%, (20.92±5.18)%, (25.29±5.83)%. Significant difference was found between five groups ( F=3.513, P=0.021). And there were significant differences between group E and A ( t=2.315, P=0.029), group E and C ( t=2.609, P=0.015). Conclusions:When cell culture time exceeded 12 h, 1% oxygen condition induced higher proliferation, lower apoptosis, and higher migration ability than 5% or 21% oxygen conditions in cultured chondrocytes. Moreover, oxygen concentration had stronger influence on the biological characteristics of rabbit auricular chondrocytes compared to intervention time.
