Effect of silane coupling agent composite graphene oxide coating on biological behavior of Schwann cells on pure titanium

doi:10.12307/2023.555

Abstract

Abstract: BACKGROUND: The study of implant surface treatment pays more attention to whether it can improve the efficiency of implant osseointegration, while the study of oral implant bone perception function is relatively less.
OBJECTIVE: To investigate the effect of silane coupling agent covalently bonded graphene oxide coating on the biological behavior of Schwann cells in vitro.
METHODS: Different mass concentrations of graphene oxide coating (0.25, 0.5, 0.75, and 1.0 mg/mL, named as 0.25-Ti-APTES-GO group, 0.5-Ti-APTES-GO group, 0.75-Ti-APTES-GO, and 1.0-Ti-APTES-GO group) were prepared on the surface of TA4 titanium sheet by covalent bonding of amino groups of silane coupling agent. The microstructure, composition and contact angle of the coating were characterized by scanning electron microscopy, Raman spectroscopy and contact instrument. Rat Schwann cells were cultured on smooth titanium sheet, silane coupling modified titanium sheet and graphene oxide coating modified titanium sheet with different mass concentrations. The proliferation activity of Schwann cells in each group was detected by CCK-8 cell proliferation assay. RT-qPCR and western blot assay were used to detect the ability of Schwann cells to secrete nerve growth factor and glial cell-derived neurotrophic factor on titanium sheets.
RESULTS AND CONCLUSION: (1) Scanning electron microscopy showed that silane coupling agent composite with different concentrations of graphene oxide coatings were cloudy, and high concentration of graphene oxide coating appeared more dense cloudy protrusions. (2) Compared with the smooth titanium sheet group, the contact angle of the silane coupling agent film titanium group was smaller (P < 0.05). The contact angle of silane coupling agent composite graphene oxide coating became small with the increase of graphene oxide concentration. Among them, the contact angle of 0.75-Ti-APTES-GO group and 1.0-Ti-APTES-GO group was smaller than that of smooth titanium sheet group (P < 0.05). (3) In the CCK-8 cell proliferation assay, compared with the smooth titanium sheet group, there was no significant change in cell proliferation in silane coupling modified titanium sheet group, 0.25-Ti-APTES-GO group and 0.5-Ti-APTES-GO group (P > 0.05). Cell proliferation was inhibited in 0.75-Ti-APTES-GO group and 1.0-Ti-APTES-GO group (P < 0.05), so 0.25-Ti-APTES-GO group and 0.5-Ti-APTES-GO group were selected for subsequent experiments. (4) RT-qPCR exhibited that the expression of nerve growth factor mRNA in 0.5-Ti-APTES-GO group was higher than that in the other three groups (P < 0.05). The expression of glial cell-derived neurotrophic factor mRNA in 0.25-Ti-APTES-GO group was lower than that in other three groups (P < 0.05). The expression of glial cell-derived neurotrophic factor mRNA in 0.5-Ti-APTES-GO group was higher than that in smooth titanium sheet group and silane coupling modified titanium sheet group (P < 0.05). (5) Western blot assay exhibited that the expression level of nerve growth factor protein in the 0.5-Ti-APTES-GO group was higher than that in the smooth titanium sheet group and 0.25-Ti-APTES-GO group (P < 0.05), and the expression level of glial cell-derived neurotrophic factor protein was higher than that in the other three groups (P < 0.05). (6) The results indicated that silane coupling agent combined with 0.5 mg/mL graphene oxide coating had good biocompatibility and could promote the expression of Schwann cell nerve growth factor and glial cell-derived neurotrophic factor on pure titanium surface.

Key words: pure titanium surface, graphene oxide, silane coupling agent, Schwann cell, bone perception, neurotrophic factor

CLC Number:

Xu Xingxing, Wen Chaoju, Li Ying, Meng Maohua, Wang Qinying, Chen Jingqiao, Dong Qiang. Effect of silane coupling agent composite graphene oxide coating on biological behavior of Schwann cells on pure titanium[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(30): 4783-4789.

Figures/Tables 7

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