Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (22): 3504-3509.doi: 10.3969/j.issn.2095-4344.2295

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Biological behaviors of fibroblasts on the surface of TiO2 nanotubes with different diameters after annealing treatment

Li Hongcai, Ma Zhuang, Guo Youling   

  1. First Department of Stomatology, Zhuzhou Central Hospital, Zhuzhou 410002, Hunan Province, China
  • Received:2019-11-12 Revised:2019-11-16 Accepted:2020-01-02 Online:2020-08-08 Published:2020-04-26
  • Contact: Ma Zhuang, Chief physician, First Department of Stomatology, Zhuzhou Central Hospital, Zhuzhou 410002, Hunan Province, China
  • About author:Li Hongcai, Master, Attending physician, First Department of Stomatology, Zhuzhou Central Hospital, Zhuzhou 410002, Hunan Province, China
  • Supported by:
    Project of Hunan Provincial Health Department, No. c2013-051

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Abstract:

BACKGROUND: Previous studies have shown that titanium nanotubes with different diameters affect the adhesion and growth of cells on the implant surface.

OBJECTIVE: To investigate the effects of annealing treatment on the biological behaviors of fibroblasts on the surface of TiO2 nanotubes with different diameters.

METHODS: TiO2 nanotubes were prepared by polishing pure titanium samples through anodizing at 5 V and 20 V respectively, and then annealed. The pure titanium samples were divided into six groups: P (polishing titanium), NT5 (TiO2 nanotubes prepared at 5 V), NT20 (TiO2 nanotubes prepared at 20 V), A-P (annealing treatment and polishing titanium), A-NT5 (TiO2 nanotubes prepared at 5V and annealed) and A-NT20 (TiO2 nanotubes formed at 20 V and annealed). Field emission scanning electron microscopy was used to obsesrve the surface topography. Fibroblasts were inoculated on the surface of each group of samples. After 60 and 120 minutes of culture, the number of cells adhering to titanium sample surface was counted using cell nucleus staining method. On day 1 of culture, cell morphology was observed using scanning electron microscopy. On days 1, 3 and 5 of culture, cell proliferation was detected by MTT method. On day 3 of culture, the secretion of collagen fiber was detected by picro-sirius red staining method.

RESULTS AND CONCLUSION: (1) The annealing treatment had no significant effect on the morphology and diameter of the nanotubes. (2) The number of adherent cells in the NT5 and NT20 groups was significantly lower than that in the P group. The annealing treatment increased the number of fibroblasts on the surface of polished pure titanium and decreased the number of fibroblasts in the NT5 and NT20 groups. (3) Annealing enhanced the viability of fibroblasts on the surface of polished pure titanium and decreased the viability of cells on the surface of nanotubes prepared at 5 and 20 V. (4) Cell viability in the NT5 and NT20 groups was lower than that in the P group. Annealing treatment increased the viability of cells on the surface of polished pure titanium and decreased the viability of cells on the surface of nanotubes prepared at 5 and 20 V. (5) The level of collagen on the surface of nanotubes in the NT5 and NT20 groups was higher than that in the P group. Annealing treatment increased the level of collagen on the surface of polished pure titanium and decreased the level of collagen in the NT5 and NT20 groups. These findings suggest that TiO2 nanotubes inhibit the adhesion, spreading and proliferation of fibroblasts to different degrees; annealing treatment can enhance this inhibitory effect; TiO2 nanotubes enhance collagen secretion by fibroblasts to different degrees, and annealing treatment inhibits the enhancement.

Key words: TiO2, nanotubes, annealing treatment, fibroblasts, cell adhesion, cell proliferation, cell spreading, collagen secretion

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