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Review
. 2013 Mar 7;14(1):015009.
doi: 10.1088/1468-6996/14/1/015009. eCollection 2013 Feb.

In vitro and in vivo evaluation of electrospun PCL/PMMA fibrous scaffolds for bone regeneration

So-Ra Son  1 , Nguyen-Thuy Ba Linh  1 , Hun-Mo Yang  2 , Byong-Taek Lee  1
Affiliations
Review

In vitro and in vivo evaluation of electrospun PCL/PMMA fibrous scaffolds for bone regeneration

So-Ra Son et al. Sci Technol Adv Mater. .

Abstract

Scaffolds were fabricated by electrospinning using polycaprolactone (PCL) blended with poly(methyl methacrylate) (PMMA) in ratios of 10/0, 7/3, 5/5 and 3/7. The PCL/PMMA ratio affected the fiber diameter, contact angle, tensile strength and biological in vitro and in vivo properties of the scaffolds, and the 7/3 ratio resulted in a higher mechanical strength than 5/5 and 3/7. In vitro cytotoxicity and proliferation of MG-63 osteoblast cells on these blended scaffolds were examined by MTT assay, and it was found that PCL/PMMA blends are suitable for osteoblast cell proliferation. Confocal images and expression of proliferating cell nuclear antigen confirmed the good proliferation and expression of cells on the 7/3 PCL/PMMA fibrous scaffolds. In vivo bone formation was examined using rat models, and bone formation was observed on the 7/3 PCL/PMMA scaffold within 2 months. In vitro and in vivo results suggest that 7/3 PCL/PMMA scaffolds can be used for bone tissue regeneration.

Keywords: PCL; PMMA; bone regeneration; electrospinning.

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Figures

Figure 1.
Figure 1.
SEM images of fibrous scaffolds electrospun at PCL/PMMA ratios of 10/0 (a), 7/3 (b), 5/5 (c) and 3/7 (d) and the corresponding diameter distributions (insets).
Figure 2.
Figure 2.
Water contact angles for 10/0, 7/3, 5/5 and 3/7 PCL/PMMA scaffolds (∗p < 0.05 versus 10/0 fibrous mats).
Figure 3.
Figure 3.
Stress and strain values of electrospun PCL/PMMA scaffolds (✦✦✦p < 0.001 versus strain value of 3/7 mats and formula image versus stress value of 3/7 scaffold).
Figure 4.
Figure 4.
Cell viability of PCL/PMMA (10/0, 7/3, 5/5 and 3/7) scaffolds measured by MTT assay (∗p < 0.05 versus 10/0 mats, ∗∗p < 0.01 versus 10/0 mats and ∗∗∗p < 0.001 versus 10/0 mats).
Figure 5.
Figure 5.
Optical density value of MG-63 cells proliferated concentration on PCL/PMMA (10/0 and 7/3) scaffolds incubated for 1, 5 and 7 days as measured by MTT assay (∗p < 0.05 versus 10/0 scaffold and ∗∗∗p < 0.001 versus culture times (1 day)).
Figure 6.
Figure 6.
Confocal microscopy images of 7/3 PCL/PMMA scaffolds (a-1) and (b-1) and MG-63 cells attached after 1 day of culture: low magnification (a-2) and (a-3) and high magnification (b-2) and (b-3). Phalloidin (green) was applied for staining membrane and DAPI (red) for staining nuclei.
Figure 7.
Figure 7.
Confocal microscopy images of MG-63 cells proliferated on the 10/0 (a-1), (a-2) and (a-3) and 7/3 PCL/PMMA scaffolds (b-1), (b-2) and (b-3) cultured for 1 day (a-1) and (b-1), 5 days (a-2) and (b-2) and 7 days (a-3) and (b-3).
Figure 8.
Figure 8.
Identification of proliferating cell nuclear antigen (PCNA) on the PCL/PMMA (10/0 and 7/3) scaffolds for 1, 5 and 7 days of incubation obtained by western blot analysis.
Figure 9.
Figure 9.
Rat skull defect without (a) and with (b) 7/3 PCL/PMMA scaffold 1 month (a-1), (b-1) and 2 months (a-2), (b-2) after implantation. The scaffold resulted in bone regeneration, whereas only minimal healing is observed for the unimplanted defect (a-1), (a-2) as evaluated by micro-CT analysis. (c) Bone volume (BV/TV) and surface density (BS/TV) for each rat with error bars showing the standard deviation (∗p < 0.05 versus BS/TV of 7/3 scaffold (2 months) and ∗∗∗p < 0.001 versus BV/TV of 7/3 scaffold (2 months)).
Figure 10.
Figure 10.
Histological sections of rat skull defects (4 × magnification, H&E staining): without implantation, after 1 month (a-1) and 2 months (a-2), and with an implanted 7/3 PCL/PMMA scaffold, after 1 month (b-1) and 2 months (b-2).
Figure 11.
Figure 11.
Histological sections of rat skull defects (40 × magnification, H&E staining): without implantation, after 1 month (a-1) and 2 months (a-2), and with an implanted 7/3 PCL/PMMA scaffold, after 1 month (b-1) and 2 months (b-2). Arrows indicate the new bone (NB), PCL/PMMA fiber (FB) and osteocyte (OC).
Figure 12.
Figure 12.
Histological sections of rat skull defects (40 × magnification, Masson's trichrome staining): without implantation, after 1 month (a-1) and 2 months (a-2), and with an implanted 7/3 PCL/PMMA scaffold, after 1 month (b-1) and 2 months (b-2). Arrows indicate collagen stained blue (Col), red-stained new bone (NB), PCL/PMMA fiber (FB) and residual scaffold construct.
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