Retinoic acid –loaded core-shell fibrous scaffold for neuronal differentiation of trabecular mesenchymal stem cells

Document Type : Research Paper

Authors

1 Department of Medical Nanotechnology, Zanjan University of Medical Sciences, Zanjan, Iran

2 Department of Medical Biotechnology, Zanjan University of Medical Sciences, Zanjan, Iran

3 Zanjan Metabolic Diseases Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

4 Zanjan Pharmaceutical Nanotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran

Abstract

Objective(s): Scientists believe that they can fabricate a biochemical scaffold and seed stem cells on it to create an extracellular matrix for tissue generation. This study sought to develop retinoic acid (RA)-loaded core-shell fibrous scaffolds (Poly-Caprolactone (PCL)/Polyethylene Oxide (PEO) based on electrospinning technique, to examine neural differentiation of trabecular mesenchymal stem cells (TM-MSCs).
Materials and Methods: PEO-PCL core- shell fibrous scaffold was fabricated using coaxial electrospinning and Fourier transform infrared (FTIR) used to evaluate the chemical bond structure, scanning electron microscopy (SEM) has been utilized to evaluate surface topography and fibrous diameter, and transient electron microscopy (TEM) to evaluate core-shell structure. The neural differentiation was evaluated using Real-Time PCR.
Results: The results of FTIR, SEM, and TEM confirm the fabrication of core-shell fibrous of PEO-PCL. The fabricated scaffold provides a suitable substrate for adhesion, cell proliferation, and differentiation. SEM images show changes in the morphology of TM-MSCs to neuronal cells. A sustained release of RA from the PEO/PCL scaffold was detected over 14 days. In addition, quantifying the expression of the gene indicates an increase in the gene expression of microtubule-associated protein 2 (MAP-2) gene.
Conclusion:The PEO/PCL core-shell fibrous scaffold containing a RA constructed using coaxial electrospinning technique was a suitable substrate for inducing neuronal differentiation of TM-MSCs cultivated on core-shell scaffold.

Keywords


1.Freed LE, Vunjak-Novakovic G, Biron RJ, Eagles DB, Lesnoy DC, Barlow SK. Biodegradable polymer scaffolds for tissue engineering. Nat Biotechnol. 1994; 12(7): 689-693.
2.Haoqing C, Ting L, Sing YC. The application of nanofibrous scaffolds in neural tissue engineering. Advanced Drug Delivery Reviews. 2009; 61(12): 1055-1064.
3.Ji W, Sun Y, Yang F, van den Beucken JJ, Fan M, Chen Z. Bioactive electrospun scaffolds delivering growth factors and genes for tissue engineering applications. Pharmaceutical research. 2011; 28(6): 1259-1272.
4.Liao S, Li B, Ma Z, Wei H, Chan C, Ramakrishna S. Biomimetic electrospun nanofibers for tissue regeneration. Biomed Mater. 2006;1.
5.Xie J, MacEwan MR, Schwartz AG, Xia Y. Electrospun nanofibers for neural tissue engineering. Nanoscale. 2010; 2(1): 35-44.
6.Elahi F, Lu W, Guoping G, Khan F. Core-shell fibers for biomedical applications-a review. Bioeng Biomed Sci J. 2013; 3(01): 1-14.
7.Ang HY, Irvine SA, Avrahami R, Sarig U, Bronshtein T, Zussman E, et al. Characterization of a bioactive fiber scaffold with entrapped HUVECs in coaxial electrospun core-shell fiber. Biomatter. 2014; 4(1): e28238.
8.Sun Z, Zussman E, Yarin AL, Wendorff JH, Greiner A. Compound core–shell polymer nanofibers by co‐electrospinning. Advanced materials. 2003; 15(22): 1929-1932.
9.Jamali S, Mostafavi H, Barati G, Eskandari M, Nadri S. Differentiation of mesenchymal stem cells -derived trabecular meshwork into dopaminergic neuron-like cells on nanofibrous scaffolds. Biologicals : journal of the International Association of Biological Standardization. 2017; 50: 49-54.
10.Nadri S, Yazdani S, Arefian E, Gohari Z, Eslaminejad MB, Kazemi B. Mesenchymal stem cells from trabecular meshwork become photoreceptor-like cells on amniotic membrane. Neuroscience Letters. 2013; 541: 43-48.
11.Dai Z, Ronholm J, Tian Y, Sethi B, Cao X. Sterilization techniques for biodegradable scaffolds in tissue engineering applications. Journal of tissue engineering. 2016; 7: 1-13.
12.Jedari B, Rahmani A, Naderi M, Nadri S. MicroRNA-7 promotes neural differentiation of trabecular meshwork mesenchymal stem cell on nanofibrous scaffold. Journal of cellular biochemistry. 2020; 121(4): 2818-2827.
13.Wu J, Xie L, Lin WZY, Chen Q. Biomimetic nanofibrous scaffolds for neural tissue engineering and drug development. Drug Discovery Today. 2017; 22(9): 1375-1384.
14.Pedersbæk D, Tudsborg Frantzen M, Fojan P. Electrospinning of Core-Shell Fibers for Drug Release Systems. 2017; 5(1): 17-30.
15.Nadri S, Nasehi F, Barati G. Effect of parameters on the quality of core-shell fibrous scaffold for retinal differentiation of conjunctiva mesenchymal stem cells. Journal of Biomedical Materials Research Part A. 2017; 105(1): 189-197.
16.Singh R, Ahmed F, Polley P, Giri J. Fabrication and Characterization of Core–Shell Nanofibers Using a Next-Generation Airbrush for Biomedical Applications. ACS Applied Materials & Interfaces. 2018; 10(49): 41924-41934.
17.Zhu X, Ohtsubo M, Böhmer RM, Roberts JM, Assoian RK. Adhesion-dependent cell cycle progression linked to the expression of cyclin D1, activation of cyclin E-cdk2, and phosphorylation of the retinoblastoma protein. The Journal of cell biology. 1996; 133(2): 391-403.
18.Howe AK, Aplin AE, Juliano RL. Anchorage-dependent ERK signaling--mechanisms and consequences. Current opinion in genetics & development. 2002; 12(1): 30-35.
19.Yang F, Xu CY, Kotaki M, Wang S, Ramakrishna S. Characterization of neural stem cells on electrospun poly(L-lactic acid) nanofibrous scaffold. Journal of biomaterials science Polymer edition. 2004; 15(12): 1483-1497.
20.Oliveira MR. The neurotoxic effects of vitamin A and retinoids. Anais da Academia Brasileira de Ciencias. 2015; 87(2 Suppl): 1361-1373.
21.Damanik FF, van Blitterswijk C, Rotmans J, Moroni L. Enhancement of synthesis of extracellular matrix proteins on retinoic acid loaded electrospun scaffolds. Journal of Materials Chemistry B. 2018; 6(40): 6468-6480.
22.Jiang X, Cao HQ, Shi LY, Ng SY, Stanton LW, Chew SY. Nanofiber topography and sustained biochemical signaling enhance human mesenchymal stem cell neural commitment. Acta biomaterialia. 2012; 8(3): 1290-1302.
23.Tiwari SK, Tzezana R, Zussman E, Venkatraman SS. Optimizing partition-controlled drug release from electrospun core-shell fibers. International journal of pharmaceutics. 2010; 392(1-2): 209-217.
24.Gonçalves A, Estevinho BN, Rocha F. Characterization of biopolymer-based systems obtained by spray-drying for retinoic acid controlled delivery. Powder Technology. 2019; 345: 758-765.
25.Korzhevskii D, Karpenko M, Kirik O. Microtubule-associated proteins as indicators of differentiation and the functional state of nerve cells. Neuroscience and Behavioral Physiology. 2012; 42(3): 215-222.
26.Laferrière NB, Brown DL. Expression and posttranslational modification of class III beta-tubulin during neuronal differentiation of P19 embryonal carcinoma cells. Cell motility and the cytoskeleton. 1996; 35(3): 188-199.