铜离子对牙髓组织再生能力的影响及其机制初探

doi:10.13591/j.cnki.kqyx.2023.05.004

Abstract

Abstract:

Objective To study the effect of copper ions on the dental pulp regenerative ability of dental pulp stem cells (DPSCs) and its primary mechanism. Methods After DPSCs were stimulated with culture medium containing copper in different concentrations, CCK-8 assay was employed for evaluating the proliferation of DPSCs and toxic effects on DPSCs; Calcein-AM/PI kit was used to test the cytotoxicity of hydrogel; Western blotting and qPCR was used to detect the protein and mRNA expression of hypoxia inducible factor-1α (HIF-1α), chromobox protein homolog 7 (CBX7) and stemness-related indicators; the phenotype of angiogenesis was examined by human umbilical vein endothelial cells (HUVECs) tubes formation assay and migration assay; the copper-containing hydrogel loaded with DPSCs was prepared and injected into the subcutaneous of nude mice for two weeks, and then hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining were used to observe the self-renewal and angiogenesis of DPSCs under the skin. Results The in vitro results showed that 5, 25, 50 μmol/L copper ions could promote the proliferation of DPSCs and had no significant toxic effects on DPSCs; Western blotting and qPCR results displayed the protein and mRNA levels of HIF-1α, CBX7, Sox-2, Oct-4 and Nanog in 25 μmol/L copper ions group were higher than that in the control group. Copper ions (25 μmol/L) could improve the tube formation and migration of HUVECs. The in vivo results showed that the copper-containing hydrogel loaded with DPSCs could lead to angiogenesis and increase dramatically the expression of Sox-2 in the subcutaneous of nude mice. Conclusion Copper ions can activate HIF-1α, enhance the expression of CBX7, and then upregulate Sox-2, Oct-4 and Nanog to maintain the stemness of DPSCs, meanwhile, can induce the regeneration of blood vessels, which can provide theoretical and experimental foundation for the regeneration of dental pulp.

Key words: copper ions, dental pulp stem cells, dental pulp regeneration, chromobox protein homolog 7(CBX7)

CLC Number:

R781.3

WU Yu,YANG Leiting,JIANG Fei,ZHOU Zhixuan,SHEN Ming. The effect and primary mechanism of copper ions on the regenerative capability of dental pulp[J]. Stomatology, 2023, 43(5): 407-414.

Figures/Tables 6

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References 24

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引物名称		引物序列
hypoxia inducible factor-1α(HIF-1α)Forward:5'-TTCACCTGAGCCTAATAGTCC-3'		Reverse:5'-CAAGTCTAAATCTGTGTCCTG-3'
chromobox protein homolog 7(CBX7)Forward:5'- GGATGGCCCCCAAAGTACAG-3'		Reverse:5'- TATACCCCGATGCTCGGTCTC-3'
SRY-box transcription factor 2(Sox-2)Forward:5'-GGCAGAGAAGAGAGTGTTTGC-3'		Reverse:5'-GCCGCCGATGATTGTTATT-3'
Octamer-binding transcription factor 4(Oct-4)Forward:5'-TGAGAGGCAACCTGGAGAAT-3'		Reverse:5'-AACCACACTCGGACCACATC-3'
Nanog homeobox(Nanog)Forward:5'-AAGGTCCCGGTCAAGAAACAG-3'		Reverse:5'- CTTCTGCGTCACACCATTGC-3'
vascular endothelial growth factor A(VEGFA)Forward:5'-AGGGCAGAATCATCACGAAGT-3'		Reverse:5'-AGGGTCTCGATTGGATGGCA-3'
β-ActinForward:5'-TCATGAAGTGTGACGTGGACAT-3'		Reverse:5'-CTCAGGAGGAGCAATGATCTTG-3'

The effect and primary mechanism of copper ions on the regenerative capability of dental pulp

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