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. 2017 Nov 15:12:8277-8287.
doi: 10.2147/IJN.S144528. eCollection 2017.

Fabrication of large-pore mesoporous Ca-Si-based bioceramics for bone regeneration

Deliang Zeng  1   2 , Xingdi Zhang  3 , Xiao Wang  1   2 , Lingyan Cao  1 , Ao Zheng  1   2 , Jiahui Du  1   2 , Yongsheng Li  3 , Qingfeng Huang  1 , Xinquan Jiang  1   2
Affiliations

Fabrication of large-pore mesoporous Ca-Si-based bioceramics for bone regeneration

Deliang Zeng et al. Int J Nanomedicine. .

Abstract

Our previous study revealed that mesoporous Ca-Si-based materials exhibited excellent osteoconduction because dissolved ions could form a layer of hydroxycarbonate apatite on the surface of the materials. However, the biological mechanisms underlying bone regeneration were largely unknown. The main aim of this study was to evaluate the osteogenic ability of large-pore mesoporous Ca-Si-based bioceramics (LPMSCs) by alkaline phosphatase assay, real-time PCR analysis, von Kossa, and alizarin red assay. Compared with large-pore mesoporous silica (LPMS), LPMSCs had a better effect on the osteogenic differentiation of dental pulp cells. LPMSC-2 and LPMSC-3 with higher calcium possessed better osteogenic abilities than LPMSC-1, which may be related to the calcium-sensing receptor pathway. Furthermore, the loading capacity for recombinant human platelet-derived growth factor-BB was satisfactory in LPMSCs. In vivo, the areas of new bone formation in the calvarial defect repair were increased in the LPMSC-2 and LPMSC-3 groups compared with the LPMSC-1 and LPMS groups. We concluded that LPMSC-2 and LPMSC-3 possessed both excellent osteogenic abilities and satisfactory loading capacities, which may be attributed to their moderate Ca/Si molar ratio. Therefore, LPMSCs with moderate Ca/Si molar ratio might be potential alterative grafts for craniomaxillofacial bone regeneration.

Keywords: dental pulp cells; mesoporous Ca-Si-based materials; rat calvarial defect.

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Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
The characterization of LPMS and LPMSC scaffolds. The proposed mechanism for the preparation of LPMS (A). Photograph of LPMSC with different shapes (B and C). Reverse color photograph of LPMSC (D). Scanning electron microscopy image of LPMSC (E). Wide-angle XRD pattern of LPMS (F). Small-angle XRS patterns of LPMS (G). Pore size distribution curves of LPMS and LPMS-cal (H). FT-IR spectrum of LPMS and LPMS-cal (I). TG curves of LPMS and LPMS-cal (J). Abbreviations: FT-IR, Fourier transform infrared; LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics; TEOS, tetraethyl orthosilicate; TG, thermogravimetric; XRD, X-ray diffraction; XRS, X-ray Raman scattering.
Figure 2
Figure 2
The characterization and cytotoxicity of LPMSCs. XPS full spectrum of different samples (A). EDS analysis for LPMSC-1 (B), LPMSC-2 (C), and LPMSC-3 (D). SEM (E–H) and fluorescence (I–L) images of DPCs on different samples after culturing for 48 hours. Abbreviations: CPS, counts per second; DPCs, dental pulp cells; LPMSC, large-pore mesoporous Ca-Si-based bioceramics; EDS, energy-dispersive spectrometry; SEM, scanning electron microscopy; XPS, X-ray photoelectron spectroscopy.
Figure 3
Figure 3
Proliferation and osteogenic differentiation of DPCs seeded on scaffolds. Proliferation of DPCs seeded on scaffolds different samples (A). ALP, von Kossa, and ARS staining photographs of DPCs seeded on scaffolds (B). Gene expression of Runx2, Osterix, Ocn, Vegf, Pparγ, and Dmp-1 for DPCs seeded on different samples after 7 days as determined by real-time PCR analysis (C). *p<0.05 vs control; #p<0.05 vs LPMS. Abbreviations: ALP, alkaline phosphatase; ARS, alizarin red S; DPCs, dental pulp cells; LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics.
Figure 4
Figure 4
Osteogenic differentiation of DPCs cultured in biomaterial extracts. ALP activity (A) and mineral deposition (B) of DPCs cultured in biomaterial extracts. ALP and ARS staining photographs of DPCs cultured in biomaterial extracts (C). Gene expression of Runx2, Osterix, Bmp-2, Dspp, Dmp-1, Opg, and Rankl for DPCs cultured in different extracts after 7 days as determined by real-time PCR analysis (D). *p<0.05 vs control; #p<0.05 vs LPMS. Abbreviations: ALP, alkaline phosphatase; ARS, alizarin red S; DPCs, dental pulp cells; LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics.
Figure 5
Figure 5
The gene expression of CaSR (A), Bmp-2 (B), Alp (C), and Ocn (D) for DPCs cultured in LPMSC extracts with NPS2143 after 7 days. *p<0.05 vs LPMSCs. Abbreviations: DPCs, dental pulp cells; LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics.
Figure 6
Figure 6
Adsorption capacity (A) and release performance (B) of rhPDGF-BB from LPMS and LPMSCs. Abbreviations: LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics; rhPDGF-BB, recombinant human platelet-derived growth factor-BB.
Figure 7
Figure 7
Histological and histomorphometric observations 8 weeks after the operation. Pathological changes in the kidney, liver, spleen, and lung (A). Different samples were stained with H&E (yellow arrow: new bone) (B) and van Gieson’s picro-fuchsin (C). Histomorphometric analysis of the new bone area (D). #p<0.05 vs LPMS; *p<0.05 vs LPMSC-1. Abbreviations: LPMS, large-pore mesoporous silica; LPMSC, large-pore mesoporous Ca-Si-based bioceramics.
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