PCL-SF/HA/GRGD cardiac patches promoting micromass formations and cardiomyogenesis of hBMSC
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1
National Yang-Ming University, Biomedical Engineering, Taiwan
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2
National Yunlin University of Sci. and Tech, Chemical Engineering, Taiwan
The interactions of hyaluronic acid (HA) and CD44 receptors of human bone marrow-derived mesenchymal stem cell (hBMSC) can promote cardiomyogenesis of the cells on silk fibroin (SF)[1], and GRGD with αvβ3 integrin ligands can enhance the cell adhesion to SF surfaces[2], respectively. Whether combinations of HA and GRGD on SF surfaces would have synergetic effects on promoting cardiomyogenesis of hBMSC has not been reported. Poly (ε-caprolactone) (PCL), PCL-SF, PCL-SF/HA and PCL-SF/HA/GRGD cardiac patches were fabricated and grafted by a photochemical technique. They were employed to culture hBMSC for investigating in-vitro cardiomyogenesis of the cells after they were induced by 5-aza. The presences of HA and GRGD on those surfaces were determined by FT-IR, toluidine blue-O assay (TBO), and X-ray photoelectron spectroscopy (XPS). Proliferations and cardiomyogenesis of hBMSC was determined by MTS assay, immune-fluorescence stains of cardiac proteins including CX43 and α-actin, respectively. The cardiac-specific gene expressions of Gata4, Actc1, Nkx2.5, and Tnnt2 were determined and analyzed by RT-PCR. In addition, Paxillin, an adhesive protein, and cytoskeletal-actin filament of differentiated hBMSC were stained by immunofluorescence. TBO assay for carboxyl groups of the HA polymers confirmed the presences of the polymers grafted to PCL-SF/HA/GRGD and PCL-SF/HA patches. In addition, XPS spectra indicated the presences of GRGD on PCL-SF/HA/GRGD patches. After 3 days of cultivation, proliferation of hBMSC on a PCL-SF/HA/GRGD patches was significantly higher than others by performing a MTS assay. Micromass formations of differentiated hBMSC on the PCL-SF/HA/GRGD patch were observed by confocal microscopy, which were about 250 µm width and 30 µm height (Fig. 1 (A)). Moreover, hBMSC micromass showed significantly higher expressions of the cardiac proteins and better cardiomyogenesis on PCL-SF/HA/GRGD patches than those of PCL-SF/HA and PCL-SF patches. The expressions of cardiac-specific genes for hBMSC micromass on PCL-SF/HA/GRGD patches were significantly higher than the other patches. Interestingly, the expressions of paxillin of the hBMSC micromass were higher on the PCL-SF/HA/GRGD patches than others (Fig.1 (B)). The synergetic effects of HA and GRGD on promoting micromass formations and cardiomyogenesis of hBMSC on PCL-SF/HA/GRGD patches was first reported that may have great potentials to be verified in in-vivo studies.
Fig. 1 the immunofluorescence stain of (A) CX43 (in green) (B) paxillin (in green) and actin (in red) of hBMSC micromass on the PCL-SF/HA/GRGD patch. (Blue: DAPI)
This work is founded by the National Science Council of the republic of China, Taiwan under Contract Nos. NSC-104-2221-E-010-004-MY3
References:
[1] M.C. Yang et al., Biomaterials, 2009, 30(22), 3757-3765
[2] E. S. Gil et al., Biomaterials, 2010, 31(34), 8953-8963
Keywords:
Cell Differentiation,
Tissue Regeneration,
Heart repair,
bioactive interface
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
New Frontier Oral
Topic:
Synthetic scaffolds as extracellular matrices
Citation:
Lo
H,
Chen
J and
Chung
T
(2016). PCL-SF/HA/GRGD cardiac patches promoting micromass formations and cardiomyogenesis of hBMSC.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01745
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.
*
Correspondence:
Dr. Hsin-Yu Lo, National Yang-Ming University, Biomedical Engineering, Taipei, Taiwan, catfish19930628@gmail.com
Dr. Jan-Hou Chen, National Yunlin University of Sci. and Tech, Chemical Engineering, Yunlin, Taiwan, Email1