Mo Doped Cuprorivaite: Preparation, Antibacterial and Cytocompatibility

doi:10.15541/jim20200506

Abstract

Abstract:

Copper-containing biomaterials have excellent inhibitory effect on bacteria growth by releasing copper ions at high concentration, which may have cytotoxicity at the same time. Although low concentration of copper ions has good cytocompatibility, the antibacterial activity is unsatisfactory. Therefore, it is of great significance to develop copper- containing biomaterials, which have excellent antibacterial property and good cytocompatibility. In this study, on considering the antagonistic effect between copper and molybdenum, molybdenum doped cuprorivaite (Mo-Cup) was synthesized by Sol-Gel method and its antibacterial properties and cell compatibility were evaluated by bacterial plate experiment and cell activity assay. The results showed that copper ions with high concentration (above 8.87 μg∙mL ^-1) released from Mo-Cup had a good inhibitory effect on Staphylococcus aureus. In addition, because the antagonistic effect between copper and molybdenum ions released from Mo-Cup can reduce the cytotoxicity of high concentration of copper ions, the survival rate of umbilical vein endothelial cells (HUVECs) was as high as 90% in the extracts of Mo-Cup at a high Cu ion concentration (9.65 μg∙mL ^-1). Therefore, molybdenum doping can be considered as an effective approach to reduce the cytotoxicity of copper-containing biomaterials for the development of low-toxic antibacterial biomaterials.

Key words: Cu, biomaterial, Mo, doping, cuprorivaite, bacteria inhibition, cytocompatibility

CLC Number:

TQ174

WANG Endian, CHANG Jiang. Mo Doped Cuprorivaite: Preparation, Antibacterial and Cytocompatibility[J]. Journal of Inorganic Materials, 2021, 36(7): 738-744.

Figures/Tables 8

Fig. 1 Properties of Mo doped cuprorivaite (Mo-Cup) (a) XRD patterns, SEM images of (b) 5Mo-Cup and (c) 10Mo-Cup, and content of (d) copper ion, (e) molybdenum ion and (f) silicon ion released from Cup, 5Mo-Cup and 10Mo-Cup in bacterial culture medium (BCM) and cell culture medium (ECM)

Fig. 2 (a,g) SEM images of (a-f) 5Mo-Cup and (g-l) 10Mo-Cup and (b-f, h-l) corresponding elemental maps (b, h): Si; (c, i): Ca; (d, j): O; (e, k): Cu; (f, l): Mo

Fig. 3 Effect of different concentrations of copper ion (CuSO4·5H2O) solution and molybdenum ion ((NH4)6Mo7O24·4H2O) solution on Staphylococcus aureus

Fig. 4 Effects of culture medium containing 10 μg?mL-1 copper ion, molybdenum ion and copper-molybdenum ions on cell activity assay (a) Effects of molybdenum ions on the proliferation of human dermal fibroblast (HDFs) and human umbilical vein endothelial cells (HUVECs)(xMo stands for medium containing x μg?mL-1 molybdenum ion; x=2, 4, 6, 8, 10); (b) Effects of copper molybdenum ions combination on the proliferation of HDFs and HUVECs (10Cu+yMo stands for medium containing 10 μg?mL-1 copper ions and y μg?mL-1 molybdenum ions; y=2, 4, 6, 8, 10). **: p<0.01

Fig. 5 Inhibition effect of extract from 10Mo-Cup bacterial culture medium on Staphylococcus aureus (a) Number of Staphylococcus aureus after being treated with different dilutions of 10Mo-Cup medium extract; (b) Related antibacterial rate. *: p<0.05; **: p<0.01

Table 1 Ion contents in the extract with 10Mo-Cup bacterial culture medium after different dilutions

Dilutions of the extract	Cu ion/ (μg∙mL^-1)	Si ion/ (μg∙mL^-1)	Mo ion/ (μg∙mL^-1)
1	709.86	88.71	243.10
1/40	17.75	2.22	6.08
1/80	8.87	1.11	3.04
1/160	4.44	0.55	1.52

Fig. 6 Effects of 10Mo-Cup culture medium extracts with different concentrations on (a) HDFs and (b) HUVECs activity assay. **: p<0.01

Table 2 Ion contents in the extract with 10Mo-Cup cell culture medium after different dilutions

Dilution of the extract	Cu ion/ (μg∙mL^-1)	Si ion/ (μg∙mL^-1)	Mo ion/ (μg∙mL^-1)
1	154.35	78.85	268.57
1/4	38.59	19.71	67.14
1/8	19.29	9.86	33.57
1/16	9.65	4.93	16.79

References 19

[1]	LI P L, HAN F X, CAO W W, et al. Carbon quantum dots derived from lysine and arginine simultaneously scavenge bacteria and promote tissue repair. Applied Materials Today, 2020,6(19):100601.
[2]	ZHANG Y, CHANG M L, BAO F, et al. Multifunctional Zn doped hollow mesoporous silica/polycaprolactone electrospun membranes with enhanced hair follicle regeneration and antibacterial activity for wound healing. Nanoscale, 2019,11(13):6315-6333. DOI URL
[3]	NING C, WANG X, LI L, et al. Concentration ranges of antibacterial cations for showing the highest antibacterial efficacy but the least cytotoxicity against mammalian cells: implications for a new antibacterial mechanism. Chemical Research in Toxicology, 2015,28(9):1815-1822. DOI URL
[4]	TAN S X, TAN S Z, LIU Y L, et al. Preparation and antibacterial property of copper-loaded activated carbon microspheres. Journal of Inorganic Materials, 2010,25(3):299-305. DOI URL
[5]	WU C T, ZHOU Y H, XU M C, et al. Copper-containing mesoporous bioactive glass scaffolds with multifunctional properties of angiogenesis capacity, osteostimulation and antibacterial activity. Biomaterials, 2013,34(2):422-433. DOI URL
[6]	TIAN T, WU C T, CHANG J. Preparation and in vitro osteogenic, angiogenic and antibacterial properties of cuprorivaite (CaCuSi₄O₁₀, Cup) bioceramics. RSC Advances, 2016,6(51):45840-45849. DOI URL
[7]	孔妮. 掺铜硅酸钙生物陶瓷的制备与表征及其促血管化性能的研究. 上海: 上海交通大学硕士学位论文, 2015.
[8]	KONG N, LIN K L, LI H Y, et al. Synergy effects of copper and silicon ions on stimulation of vascularization by copper-doped calcium silicate. Journal of Materials Chemistry B, 2014,2(8):1100-1110. DOI URL
[9]	YANG Z J, LONG T, RAN L W, et al. Molybdenum's biological function and roles in animal production. Journal of Henan University of Science and Technology (Agricultrual Science), 2004(2):40-43.
[10]	WU M J. Molybdenum and human health. Studies of Trace Elements and Health, 2006,5(23):66-67.
[11]	WANG Q X. Trace element molybdenum and human health. Studies of Trace Elements and Health, 2003,4(20):58-59.
[12]	LIU M. The effect of molybdenum on human health. China Molybdenum Industry, 2001,5(25):43-45.
[13]	田瑶. 四硫代钼酸铵抑制顺铂与人铜伴侣蛋白Atoxl的相互作用. 合肥: 中国科学技术大学硕士学位论文, 2018.
[14]	ALVAREZ H M, XUE Y, ROBINSON C D, et al. Tetrathiomolybdate inhibits copper trafficking proteins through metal cluster formation. Science, 2010,327(5963):331-334. DOI URL
[15]	PAN Q, KLEER C G, VAN GOLEN K L, et al. Copper deficiency induced by tetrathiomolybdate suppresses tumor growth and angiogenesis. Cancer Research, 2002,62(17):4854-4859.
[16]	CHAN N, WILLIS A, KORNHAUSER N, et al. Influencing the tumor microenvironment: a phase II study of copper depletion using tetrathiomolybdate in patients with breast cancer at high risk for recurrence and in preclinical models of lung metastases. Clinical Cancer Research, 2017,23(3):666-676. DOI URL
[17]	MIAO Z Z, LI G W, LIU C Z, et al. Study on antibacterial properties of copper-loaded chitosan particles. Journal of Henan Institute of Science and Technology, 2010,38(2):78-80.
[18]	XU Q, CHANG M L, ZHANG Y, et al. PDA/Cu bioactive hydrogel with "hot ions effect" for inhibition of drug-resistant bacteria and enhancement of infectious skin wound healing. ACS Applied Materials & Interfaces, 2020,12(28):31255-31269.
[19]	LI J, ZHAI D, LU F, et al. Preparation of copper-containing bioactive glass/eggshell membrane nanocomposites for improving angiogenesis, antibacterial activity and wound healing. Acta Biomater., 2016,36:254-266. DOI URL