Inducing Macrophages M2 Polarization by Dexamethasone Laden Mesoporous Silica Nanoparticles from Titanium Implant Surface for Enhanced Osteogenesis

doi:10.1007/s40195-019-00926-y

Abstract

Abstract:

The study conveys an idea to enhance the osseointegration of titanium implant (Ti) through modulating macrophages M2 polarization. The ~ 100 nm spherical mesoporous silica nanoparticles (MSN) that compromised of ~ 4-nm-diameter nano-tunnels were synthesized by the conventional “sol-gel” method, into which the dexamethasone (DEX) was loaded (DEX@MSN). The DEX@MSN could consistently release DEX and showed favorable cytocompatibility in RAW264.7 cells. The arginase-1 expression, a specific marker for macrophages M2 polarization, was also enhanced by DEX@MSN treatment. Then, the Ti was pre-treated with anodization under 5 V to generate the titania nanotubes with ~ 30 nm diameter (NT-30) and the DEX@MSN was introduced onto NT-30 surface via electrophoretic deposition, with the aid of chitosan. After optimizing the deposition parameters, the supernatants of RAW264.7 from the decorated implant surface could significantly promote the osteogenic differentiation of murine primary bone marrow mesenchymal stem cells. These findings demonstrate that delivery of DEX from implant surface can modulate the macrophages M2 polarization and result in favorable osteogenesis.

Key words: Macrophages polarization, Titanium implant, Osteogenesis, Dexamethasone, Mesoporous silica nanoparticles, Electrophoretic deposition

Jing Luo, Xin Ding, Wen Song, Jian-Ying Bai, Jing Liu, Zhe Li, Fan-Hui Meng, Fang-Hao Chen, Yu-Mei Zhang. Inducing Macrophages M2 Polarization by Dexamethasone Laden Mesoporous Silica Nanoparticles from Titanium Implant Surface for Enhanced Osteogenesis[J]. Acta Metallurgica Sinica (English Letters), 2019, 32(10): 1253-1260.

Figures/Tables 5

Fig. 1 Illustration of experimental design. The DEX was loaded into MSN to formulate the DEX@MSN particles, which were immobilized onto pre-anodized Ti implant surface through EPD. The functionalized Ti implant was able to induce macrophages M2 polarization, which in turn contributed to BMSCs osteogenic differentiation

Fig. 2 Characterizations of MSN and DEX@MSN. a The lateral view of MSN under TEM. b The frontal view of MSN under TEM. c The SEM image of MSN. d The surface height image of MSN by AFM scanning. e TEM image of DEX@MSN. f The TEM image of DEX@MSN. g The linear relationship between optical density (OD) values and DEX concentrations. h The cumulative released DEX percentage from DEX@MSN particles

Fig. 3 Cytotoxicity and M2 polarization induction of DEX@MSN. The RAW264.7 cells were treated with various concentrations of MSN, and the cell viability was checked by CCK-8 assay after culture for 1 day a. The RAW264.7 cells were treated with various concentrations of DEX@MSN, and the cell viability was checked by CCK-8 assay after culture for 1 and 3 days b. The RAW264.7 cells were treated with various concentrations of DEX and 2 μg/ml DEX@MSN. Total intracellular RNA was isolated to perform the qPCR analysis of Arg1 c. *P < 0.05 versus cell

Fig. 4 (DEX@MSN)Chi-Ti observation, cytotoxicity and M2 polarization induction. a The Ti implant was anodized at 5 V to formulate the NT-30 surface. Then, the EPD was performed for different times to immobilize DEX@MSN. The samples were freeze-dried and examined by SEM. b The RAW264.7 cells were cultured on (DEX@MSN)Chi-Ti surface for 1 and 3 days. The cell viability was determined by CCK-8 assay. *P < 0.05 versus cell. c The RAW264.7 cells were cultured on (DEX@MSN)Chi-Ti surface for 3 days. Total intracellular RNA was isolated to perform the qPCR analysis of Arg1. *P < 0.05 versus cell

Fig. 5 Osteogenic differentiation of BMSCs after receiving macrophages conditioned medium. The RAW264.7 cells were cultured on different surfaces, and the supernatants were collected to mix with conventional osteogenic medium to formulate the conditioned medium. The BMSCs were cultured on NT-30 surface and subjected to osteogenic differentiation in conditioned medium. The Alizarin Red staining a and ALP activity b were performed to evaluate the osteogenic differentiation efficacy. *P < 0.05

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