BSA-lysozyme coated NaCa2HSi3O9 nanorods on titanium for cytocompatibility and antibacterial activity

doi:10.1016/j.jmst.2021.01.074

Abstract

Abstract:

Extracellular supplement of Ca and Si accelerates osteogenisis and angiogenisis during bone repair. Ca-Si-based ceramics are biodegradable and can release Ca and Si ions efficiently. However, locally high alkalinity induced by excessively rapid degradation limits their application. In this paper, NaCa₂HSi₃O₉ (NCS) nanorods were fabricated on Ti by micro-arc oxidation and hydrothermal treatment; protein layers consisted of different rations of bovine serum albumin (BSA) and lysozyme (LYS) were prepared on NCS nanorods by spinning coating and thermal stabilization. The microstructure, ion releasing and wettability of coatings were observed; osteoblast and endotheliocyte behaviors, as well as bacteria response in vitro, were evaluated. The protein layer did not change microstructure and phase composition of nanorods but acted as a barrier isolating nanorods from contacting medium, and it results in slowing down the degradation of nanorods. The reduced increase of pH value, released Ca and Si ions and incorporation of LYS in BSA protein layer gave NCS nanorods improved MC3T3-E1 and HUVECs response, compared with Ti. Meanwhile, the protein layers except pure LYS endowed nanorods with a highly antibacterial effect against staphylococcus aureus and escherichia coli simultaneously, indicating the key role of BSA in resisting antibacterial adhesion. NCS nanorods with BSA and LYS composited protein layers should be a potential coating applied on Ti implants for increasing osseointegration.

Key words: NaCa₂HSi₃O₉ nanorods, Bovine serum albumin, Lysozyme, Antibacterial activity, Cytocompatibility

Yang Xue, Jun Chen, Lan Zhang, Yong Han. BSA-lysozyme coated NaCa₂HSi₃O₉ nanorods on titanium for cytocompatibility and antibacterial activity[J]. J. Mater. Sci. Technol., 2021, 88: 240-249.

Figures/Tables 8

Fig. 1. Schematic illustration of the preparation of protein-coated NCS nanorods. NCS nanorods degrade rapidly and can't support the cell well in medium with a high pH value. The protein layer consisting of BSA and LYS is expected to decrease the NCS degradation rate, reduce the pH increase and endow Ti implants with improved osteoblast/endothelial cell response and anti-bacterial activity against S. aureus and E. coli simultaneously in vitro.

Table 1 Treatment process for fabricating different surfaces.

Name	Spinning coating
Name	BSA (50 mg/mL)	LYS (25 mg/mL)	LYS (50 mg/mL)
NCS-B5	√
NCS-B5L2.5	√	√
NCS-B5L5	√		√
NCS-L5			√

Fig. 2. Surface morphologies of different samples: (a) NCS, (b) NCS-B5, (c)NCS-B5L2.5, (d)NCS-B5L5, (e)NCS-L5; (f) XRD patterns of di? ;erent samples.

Fig. 3. (a) Full XPS spectra of different coatings; (b) elemental compositions detected on different coatings by XPS; high-resolution spectra of C 1s (c) and N 1s (d) for different surfaces.

Fig. 4. (a) Contact angles and corresponding digital photographs of water droplets on different surfaces; (b) Ca and (c) Si releasing kinetics for different samples after different immersion time; (d) pH values of PS solutions with different samples after immersion for di? ;erent time; (e) surface morphologies of different surfaces after immersion in PS for 28 days, and arrows show the nanorods after serious degradation.

Fig. 5. Antibacterial ratios of S. aureus (a) and E. coil (b) on different surfaces; Live/dead staining and SEM images of S. aureus (c) and E. coil (d) cultured on different surfaces after 24 h of incubation.

Fig. 6. (a) Alamar Blue assays of cells viability after 1, 3 and 5 days incubation; (b) fluorescence images of Live/dead staining after culturing for 1 and 3 days; (c) SEM images of osteoblast cultured on different samples for 1 day; (d) gene expressions of MC3T3-E1 cultured on different samples after 5 days; (e) collagen secretion of MC3T3-E1 incubated on different samples for 5 days; (f) ECM mineralization of MC3T3-E1 incubated on different samples for 10 days. (*) p < 0.05 and (**)p < 0.01 compared with the Ti, (#) p < 0.05 and (##) p < 0.01 compared with NCS.

Fig. 7. (a) MTT assays of HUVECs cultured in 100 % coating extract for 1, 3 and 7 days, (*) p < 0.05 and (**) p < 0.01 compared with ECM; (b) fluorescence images of live/dead staining after culturing 3 days; (c) microscopy images of tube formation in vitro; quantification analysis of (d) junction and (e) master segment length, (*) p < 0.05 and (**) p<0.01 compared with ECM.

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BSA-lysozyme coated NaCa₂HSi₃O₉ nanorods on titanium for cytocompatibility and antibacterial activity

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