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. 2018 Jun;5(3):159-166.
doi: 10.1093/rb/rby009. Epub 2018 May 3.

Synthesis of photo-reactive poly (vinyl alcohol) and construction of scaffold-free cartilage like pellets in vitro

Bao Li  1 , Yongli Gao  1 , Likun Guo  1   2 , Yujiang Fan  1 , Naoki Kawazoe  2 , Hongsong Fan  1 , Xingdong Zhang  1 , Guoping Chen  2
Affiliations

Synthesis of photo-reactive poly (vinyl alcohol) and construction of scaffold-free cartilage like pellets in vitro

Bao Li et al. Regen Biomater. 2018 Jun.

Abstract

Photo-reactive poly(vinyl alcohol) (PRPVA) was synthesized by introduction of phenyl azido groups into poly(vinyl alcohol) (PVA) and applied for surface modification. PRPVA was grafted onto cell culture plate surface homogeneously or in a micropattern. Human mesenchymal stem cells (hMSCs) cultured on cell culture plate surface and PVA-modified surface showed different behaviors. Cells adhered and spread well on cell culture plate surface, while they did not adhere on PVA-grafted surface at all. When hMSCs were cultured on PVA-micropatterned surface, they formed a cell micropattern. Cells formed pellets after cultured on PVA homogeneously modified surface in chondrogenic induction medium for 2 weeks. The pellets were positively stained by hematoxylin/eosin, safranin-O/fast green and toluidin blue, and they were also stained brown by Type II collagen and proteoglycan immunohistological staining. Real-time PCR analysis was conducted to investigate the expression of colI, colII, colX, aggrecan and sox9 mRNA. Results of gene expression were in agreement with those of histological and immunohistological observations. These results indicated that hMSCs cultured on PVA-modified surface performed chondrogenic differentiation, and it was possible to construct scaffold-free cartilage like pellets with PVA-modified surface in vitro.

Keywords: chondrogenic differentiation; photo-reactive poly (vinyl alcohol); surface modification.

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Figures

Figure 1
Figure 1
1H-NMR spectra of PVA (a) and PRPVA (b) in D2O
Figure 2
Figure 2
Optical microscope images of photomask (a), PVA micropattern with low (b) and high (c) magnification. Scale bar is 500 μm for (a) and (b); 200 μm for (c)
Figure 3
Figure 3
Cells morphology cultured on PVA-modified surface in chondrogenic induction medium after 0.5 h (a), 3 h (b), 1 day (c), 2 days (d), 14 days (e) and cells morphology cultured on PVA micropatterned surface for 1 day (f)
Figure 4
Figure 4
H&E (a, d, g, j), safranin-O/fast green (b, e, h, k) and toluidin blue (c, f, i, l) staining of pellet formed on PVA-modified surface (a–f) and cell culture plate surface (g–l) cultured in chondrogenic induction medium for 2 weeks. Scale bar is 500 μm for (a–c) and (g–i); 200 μm for (d–f) and (j–l)
Figure 5
Figure 5
Immunostaining of Type I collagen (a, d, g, j), Type II collagen (b, e, h, k) and cartilage proteoglycan (c, f, i, l) of pellet formed on PVA-modified surface (a–f) and cell culture plate surface (g–l) cultured in chondrogenic induction medium for 2 weeks. Scale bar is 500 μm for (a–c) and (g–i); 200 μm for (d–f) and (j–l)
Figure 6
Figure 6
Real-time PCR results of mRNA expression of Type I collagen, Type II collagen, Type X collagen, sox9 and aggrecan of hMSCs cultured on PVA-modified surface and cell culture plate surface in the control or differentiation medium for 2 weeks. The data are normalized to GAPDH value
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References

    1. Pittenger MF, Mackay AM, Beck SC. et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999;284:143–7. - PubMed
    1. Caplan AI, Bruder SP.. Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. Trends Mol Med 2001;7:259–64. - PubMed
    1. Gonzalez-Fernandez T, Tierney EG, Cunniffe GM. et al. Gene delivery of TGF-beta3 and BMP2 in an MSC-laden alginate hydrogel for articular cartilage and endochondral bone tissue engineering. Tissue Eng A 2016;22:776–87. - PubMed
    1. Shimomura K, Ando W, Moriguchi Y. et al. Next generation mesenchymal stem cell (MSC)-based cartilage repair using scaffold-free tissue engineered constructs generated with synovial mesenchymal stem cells. Cartilage 2015;6:13S–29S. - PMC - PubMed
    1. Pan Z, Duan P, Liu X. et al. Effect of porosities of bilayered porous scaffolds on spontaneous osteochondral repair in cartilage tissue engineering. Regen Biomater 2015;2:9–19. - PMC - PubMed