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. 2017:2017:9251806.
doi: 10.1155/2017/9251806. Epub 2017 Nov 19.

Preparation of P3HB4HB/(Gelatin + PVA) Composite Scaffolds by Coaxial Electrospinning and Its Biocompatibility Evaluation

Min-Xian Ma  1   2 , Qin Liu  1   2 , Chuan Ye  3   4 , Brian Grottkau  5 , Bing Guo  1 , Yu-Feng Song  1
Affiliations

Preparation of P3HB4HB/(Gelatin + PVA) Composite Scaffolds by Coaxial Electrospinning and Its Biocompatibility Evaluation

Min-Xian Ma et al. Biomed Res Int. 2017.

Abstract

This study was conducted to prepare coaxial electrospun scaffolds of P3HB4HB/(gelatin + PVA) with various concentration ratios with P3HB4HB as the core solution and gelatin + PVA mixture as the shell solution; the mass ratios of gelatin and PVA in each 10 mL shell mixture were 0.6 g : 0.2 g (Group A), 0.4 g : 0.4 g (Group B), and 0.2 g : 0.6 g (Group C). The results showed that the pore size, porosity, and cell proliferation rate of Group C were better than those of Groups A and B. The ascending order of the tensile strength and modulus of elasticity was Group A < Group B < Group C. The surface roughness was Group C > Group B > Group A. The osteogenic and chondrogenic-specific staining showed that Group C was stronger than Groups A and B. This study demonstrates that when the mass ratio of gelatin : PVA was 0.2 g : 0.6 g, a P3HB4HB/(gelatin + PVA) composite scaffold with a core-shell structure can be prepared, and the scaffold has good biocompatibility that it may be an ideal scaffold for tissue engineering.

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Figures

Figure 1
Figure 1
General appearance of the P3HB4HB/(GEL + PVA) scaffolds. (a) GEL : PVA = 0.6 g : 0.2 g; (b) GEL : PVA = 0.4 g : 0.4 g; (c) GEL : PVA = 0.2 g : 0.6 g.
Figure 2
Figure 2
Surface roughness of the P3HB4HB/(GEL + PVA) scaffolds. (a) Roughness profile (Ra); (b) roughness height (Rz); (c) distance between contour peak line and contour bottom line (Ry). A: GEL : PVA = 0.6 g : 0.2 g; B: GEL : PVA = 0.4 g : 0.4 g; C: GEL : PVA = 0.2 g : 0.6 g. ∗∗P < 0.01.
Figure 3
Figure 3
Contact angles of the P3HB4HB/(GEL + PVA) scaffolds. (a) GEL : PVA = 0.6 g : 0.2 g; (b) GEL : PVA = 0.4 g : 0.4 g; (c) GEL : PVA = 0.2 g : 0.6 g. A: GEL : PVA = 0.6 g : 0.2 g; B: GEL : PVA = 0.4 g : 0.4 g; C: GEL : PVA = 0.2 g : 0.6 g.
Figure 4
Figure 4
Stress-strain test and biomechanical properties of the P3HB4HB/(GEL + PVA) scaffolds. (a) Stress-strain test of GEL : PVA = 0.6 g : 0.2 g. (b) Stress-strain test of GEL : PVA = 0.4 g : 0.4 g. (c) Stress-strain test of GEL : PVA = 0.2 g : 0.6 g. (d) Tensile strength. (e) Young's modulus. (f) Nominal strain fracture. A: GEL : PVA = 0.6 g : 0.2 g; B: GEL : PVA = 0.4 g : 0.4 g; C: GEL : PVA = 0.2 g : 0.6 g. ∗P < 0.05; ∗∗P < 0.01.
Figure 5
Figure 5
In vitro degradation of the P3HB4HB/(GEL + PVA) scaffolds on days 1, 3, 5, and 7 d. Group A: GEL : PVA = 0.6 g : 0.2 g; Group B: GEL : PVA = 0.4 g : 0.4 g; Group C: GEL : PVA = 0.2 g : 0.6 g. Compare to Group A; ∗P < 0.05; ∗∗P < 0.01; compare to Group B; #P < 0.05; ##P < 0.01.
Figure 6
Figure 6
Scanning electron microscopy and transmission electron microscopy of the P3HB4HB/(GEL + PVA) scaffolds. (a, d) GEL : PVA = 0.6 g : 0.2 g; (b, e) GEL : PVA = 0.4 g : 0.4 g; (c, f) GEL : PVA = 0.2 g : 0.6 g. (g, h, i) GEL : PVA = 0.2 g : 0.6 g for the scaffolds not soaked in water, soaked for 24 h, and soaked for 48 h.
Figure 7
Figure 7
Cell adhesion rate of the P3HB4HB/(GEL + PVA) scaffolds on 1, 3, and 6 h. Group A: GEL : PVA = 0.6 g : 0.2 g; Group B: GEL : PVA = 0.4 g : 0.4 g; Group C: GEL : PVA = 0.2 g : 0.6 g. Compare to Group A, ∗P < 0.05; compare to Group B, #P < 0.05.
Figure 8
Figure 8
CCK-8 assay shows the proliferation of hBMSCs on days 1, 3, 5, 7, and 9  for the scaffold materials Group A: GEL : PVA = 0.6 g : 0.2 g; Group B: GEL : PVA = 0.4 g : 0.4 g; Group C: GEL : PVA = 0.2 g : 0.6 g. Compare to Group A: ∗P < 0.05; ∗∗P < 0.01; compare to Group B: #P < 0.05; ##P < 0.01.
Figure 9
Figure 9
Observation of hBMSCs-P3HB4HB/(GEL + PVA) under electron microscope after in vitro culture for 7 days. (a) GEL : PVA = 0.6 g : 0.2 g; (b) GEL : PVA = 0.4 g : 0.4 g; (c) GEL : PVA = 0.2 g : 0.6 g.
Figure 10
Figure 10
DAPI staining of hBMSCs-P3HB4HB/(GEL + PVA) after in vitro culture for 7 days. (a) GEL : PVA = 0.6 g : 0.2 g; (b) GEL : PVA = 0.4 g : 0.4 g; (c) GEL : PVA = 0.2 g : 0.6 g. The magnification is 200x.
Figure 11
Figure 11
(a, d, g, j) GEL : PVA = 0.6 g : 0.2 g for alizarin red S, ALP, alcian blue, and safranin O staining; (b, e, h, k) GEL : PVA = 0.4 g : 0.4 g for alizarin red S, ALP, alcian blue, and safranin O staining; (c, f, i, l) GEL : PVA = 0.2 g : 0.6 g for alizarin red S, ALP, alcian blue, and safranin O staining. The magnification is 200x.
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