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. 2020 Jan 2;12(1):69.
doi: 10.3390/polym12010069.

Synergistic Effects on Incorporation of β-Tricalcium Phosphate and Graphene Oxide Nanoparticles to Silk Fibroin/Soy Protein Isolate Scaffolds for Bone Tissue Engineering

Fan Liu  1   2 , Chen Liu  1 , Bowen Zheng  2 , Jia He  2 , Jun Liu  2 , Cen Chen  3 , In-Seop Lee  4 , Xiaohong Wang  1   5 , Yi Liu  2
Affiliations

Synergistic Effects on Incorporation of β-Tricalcium Phosphate and Graphene Oxide Nanoparticles to Silk Fibroin/Soy Protein Isolate Scaffolds for Bone Tissue Engineering

Fan Liu et al. Polymers (Basel). .

Abstract

In bone tissue engineering, an ideal scaffold is required to have favorable physical, chemical (or physicochemical), and biological (or biochemical) properties to promote osteogenesis. Although silk fibroin (SF) and/or soy protein isolate (SPI) scaffolds have been widely used as an alternative to autologous and heterologous bone grafts, the poor mechanical property and insufficient osteoinductive capability has become an obstacle for their in vivo applications. Herein, β-tricalcium phosphate (β-TCP) and graphene oxide (GO) nanoparticles are incorporated into SF/SPI scaffolds simultaneously or individually. Physical and chemical properties of these composite scaffolds are evaluated using field emission scanning electron microscope (FESEM), X-ray diffraction (XRD) and attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR). Biocompatibility and osteogenesis of the composite scaffolds are evaluated using bone marrow mesenchymal stem cells (BMSCs). All the composite scaffolds have a complex porous structure with proper pore sizes and porosities. Physicochemical properties of the scaffolds can be significantly increased through the incorporation of β-TCP and GO nanoparticles. Alkaline phosphatase activity (ALP) and osteogenesis-related gene expression of the BMSCs are significantly enhanced in the presence of β-TCP and GO nanoparticles. Especially, β-TCP and GO nanoparticles have a synergistic effect on promoting osteogenesis. These results suggest that the β-TCP and GO enhanced SF/SPI scaffolds are promising candidates for bone tissue regeneration.

Keywords: graphene oxide; osteogenesis; scaffold; silk fibroin; soy protein isolate; β-tricalcium phosphate.

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Conflict of interest statement

The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
General observation of the SF/SPI-based composite scaffolds.
Figure 2
Figure 2
Surface morphology of the silk fibroin/soy protein isolate (SF/SPI)-based composite scaffolds ((A): SF; (B): SF/SPI; (C): SF/SPI/graphene oxide (GO); (D): SF/SPI/β-tricalcium phosphate (β-TCP); (E): SF/SPI/GO/β-TCP). Enlarged views of the pore structures are on the top right corner of each figures.
Figure 3
Figure 3
Mechanical properties of the SF/SPI-based composite scaffolds. Statistical significance relative to each group: * p < 0.05.
Figure 4
Figure 4
X-ray diffraction (XRD) patterns of the SF/SPI-based composite scaffolds.
Figure 5
Figure 5
ATR-FTIR patterns of the SF/SPI-based composite scaffolds.
Figure 6
Figure 6
Water adsorption behaviors of the SF/SPI-based composite scaffolds in 24 h.
Figure 7
Figure 7
Field emission scanning electron microscope (FESEM) photographs of the SF/SPI-based composite scaffolds after immersing into simulated body fluid (SBF) solution at different time points.
Figure 8
Figure 8
XRD patterns of the SF/SPI-based composite scaffolds after immersing into SBF solution for 14 days (circle refers to hydroxyapatite; arrow refers to apatite; square refers to β-TCP).
Figure 9
Figure 9
Scanning electron microscope (SEM) photographs of bone marrow mesenchymal stem cells (BMSCs) seeding on the SF/SPI-based composite scaffolds ((A): SF; (B): SF/SPI; (C): SF/SPI/GO; (D): SF/SPI/β-TCP; (E): SF/SPI/GO/β-TCP).
Figure 10
Figure 10
Proliferation of BMSCs on the SF/SPI-based scaffolds on day 1, 3, 5, and 7. Statistical significance relative to each group: * p < 0.05.
Figure 11
Figure 11
Confocal images of BMSCs on the SF/SPI-based scaffolds ((A): SF; (B): SF/SPI; (C): SF/SPI/GO; (D): SF/SPI/β-TCP; (E): SF/SPI/GO/β-TCP).
Figure 12
Figure 12
ALP activities of BMSCs on the SF/SPI-based composite scaffolds on day 1, 3, 5 and 7. Statistical significance relative to each group: * p < 0.05.
Figure 13
Figure 13
Runx 2 mRNA expression of BMSCs on the SF/SPI-based composite scaffolds on day 7 and 14. Statistical significance relative to each group: * p < 0.05.
Figure 14
Figure 14
OC mRNA expression of BMSCs on the SF/SPI-based composite scaffolds on day 7 and 14. Statistical significance relative to each group: * p < 0.05.
Figure 15
Figure 15
Col I mRNA expression of BMSCs on the SF/SPI-based composite scaffolds on day 7 and 14. Statistical significance relative to each group: *p < 0.05.
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