Applying Differentiated Retinal Cell for Age-Related Macular Degeneration Treatment

Document Type : Research Article


1 Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, I.R. IRAN

2 Department of Biotechnology, Faculty of New Sciences and Technologies, Semnan University, Semnan, I.R. IRAN

3 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology-ACECR, Tehran, I.R. IRAN

4 Bioprocess Engineering Group, Industrial and Environmental Biotechnology Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, I.R. IRAN

5 Pharmaceutical Sciences Research Center, The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Science, Tehran, I.R. IRAN


Age-related Macular Degeneration (AMD) is one of the retinal degenerative diseases associated with some degree of dysfunction and loss of Retinal Pigmented Epithelium (RPE) cells and leads to permanent sight loss. Available treatments only slow down its progression. Applying a scaffold to help RPE cells proliferation and make layers has been proposed as a promising approach to treat this group of diseases. In this study, a fuzzy system was used to optimize the situation of making a scaffold. For better adhesion and proliferation of cells, the polycaprolactone scaffold's surface was modified by alkaline hydrolysis and plasma. Some analyses, such as water uptake and biodegradation rate, were done. Then, differentiated human embryonic stem cells (hESCs) were cultured on several groups of scaffolds. Finally, the viability, proliferation, and morphology of differentiated hESC-RPE cells on all groups of the scaffolds were investigated. The nanofibers' diameter was minimized by optimizing voltage and solution concentration with a fuzzy model for the first time, which obtained 110.5 nm, 18.9 kV, and 0.065 g/mL (w/v), respectively. The immersion time of the scaffold in alkaline solution and solution concentration during surface modification were achieved 4.3 M and 104 minutes, respectively, by response surface methodology. Results of the MTT assay showed that the hydrolyzed group had a high proliferation of cells. Scanning electron microscopy observation of cell morphology after 60 days confirmed this result. In conclusion, our results demonstrate that the hydrolyzed scaffold is a suitable bed for cell proliferation, a good option for AMD treatment.


Main Subjects

[1] Kashani A.H., Lebkowski J.S., Rahhal F.M., Avery R.L., Salehi-Had H., Dang W., Lin C.M., Mitra D., Zhu D., Thomas B.B., Hikita S.T., A Bioengineered Retinal Pigment Epithelial Monolayer for Advanced, Dry Age-related Macular Degeneration, Sci. Transl. Med., 10: eaao4097 (2018).
[2] Thomas B.B., Zhu D., Zhang L., Thomas P.B., Hu Y., Nazari H., Stefanini F., Falabella P., Clegg D.O., Hinton D.R., Humayun M.S., Survival and Functionality of hESC-derived Retinal Pigment Epithelium Cells Cultured as a Monolayer on Polymer Substrates Transplanted in RCS Rats, Investig. Ophthalmol. Vis. Sci., 57: 2877-2887 (2016).
[3] Sadda S.R., Guymer R., Holz F.G., Schmitz-Valckenberg S., Curcio C.A., Bird A.C., Blodi B.A., Bottoni F., Chakravarthy U., Chew E.Y., Csaky K., Consensus Definition for Atrophy Associated with Age-Related Macular Degeneration on OCT: Classification of Atrophy Report 3, Ophthalmology, 125: 537-548 (2018).
[4] Mitchell P., Liew G., Gopinath B., Wong T.Y., Age-Related Macular Degeneration, Lancet, 392: 1147-1159 (2018).
[5] Zadeh M.A., Khoder M., Al-Kinani A.A., Younes H.M., Alany R.G., Retinal Cell Regeneration Using Tissue Engineered Polymeric Scaffolds, Drug Discov. Today, 24: 1669–1678 (2019).
[6] Hotaling N.A., Khristov V., Wan Q., Sharma R., Jha B.S., Lotfi M., Maminishkis A., Simon Jr C.G., Bharti K., Nanofiber Scaffold-Based Tissue-Engineered Retinal Pigment Epithelium to Treat Degenerative Eye Diseases, J. Ocul. Pharmacol. Ther., 32: 272–285 (2016).
[7] Di Foggia V., Makwana P., Ali R.R., Sowden J.C., Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement, J. Ocul. Pharmacol. Ther., 32: 240-252 (2016).
[8] Shahmoradi S., Hatamian A.S., Tabandeh F., Yazdian F., Polycaprolacton as a Suitable Scaffold for Retina Diseases: Based on Statistical Analysis, (2015).
[9] Mazzilli J.L., Snook J.D., Simmons K., Domozhirov A.Y., Garcia C.A., Wetsel R.A., et al., A Preclinical Safety Study of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells for Macular Degeneration, J. Ocul. Pharmacol. Ther., 36: 65–69 (2020).
[10] Song W.K., Park K.M., Kim H.J., Lee J.H., Choi J., Chong S.Y., Shim S.H., Del Priore L.V., Lanza R., Treatment of Macular Degeneration Using Embryonic Stem Cell-derived Retinal Pigment Epithelium: Preliminary Results in Asian Patients, Stem Cell Reports, 4: 860-872 (2015).
[11] Saatchi A.R., Seddiqi H., Amoabediny G., Helder M.N., Zandieh-Doulabi B., Klein-Nulend J., Computational Fluid Dynamics in 3D-Printed Scaffolds with Different Strand-Orientation in Perfusion Bioreactors, Iran. J. Chem. Chem. Eng. (IJCCE), 39: 307–320 (2020).
[12] Forest D.L., Johnson L. V., Clegg D.O., Cellular Models and Therapies for Age-Related Macular Degeneration, DMM Dis. Model. Mech., 8: 421-427 (2015).
[16] Calejo M.T., Ilmarinen T., Jongprasitkul H., Skottman H., Kellomäki M., Honeycomb Porous Films as Permeable Scaffold Materials for Human Embryonic Stem Cell-derived Retinal Pigment Epithelium, J. Biomed. Mater. Res. - Part A, (2016).
[18] Shahmoradi S., Yazdian F., Tabandeh F., Soheili Z.S., Hatamian Zarami A.S., Navaei-Nigjeh M., Controlled Surface Morphology and Hydrophilicity of Polycaprolactone Toward Human Retinal Pigment Epithelium Cells, Mater. Sci. Eng. C, 73: 300-309 (2017).
[19] Hafizi A., Koolivand-Salooki M., Janghorbani A., Ahmadpour A., Moradi M.H., An Investigation of Artificial Intelligence Methodologies in the Prediction of the Dirty Amine Flow Rate of a Gas Sweetening Absorption Column, Pet. Sci. Technol., 32: 527-534 (2014).
[20] Qin X., Wu D., Effect of Different Solvents on Poly(caprolactone)(PCL) Electrospun Nonwoven Membranes, J. Therm. Anal. Calorim., 107: 1007-1013 (2012).
[21] Baker S.R., Banerjee S., Bonin K., Guthold M., Determining the Mechanical Properties of Electrospun Poly-ε-caprolactone (PCL) Nanofibers Using AFM and a Novel Fiber Anchoring Technique, Mater. Sci. Eng. C, 59: 203-212 (2016).
[23] Jang J.S.R., ANFIS: Adaptive-Network-Based Fuzzy Inference System, IEEE Trans. Syst. Man Cybern., (1993).
[24] Chang F.J., Chang Y.T., Adaptive Neuro-fuzzy Inference System for Prediction of Water Level in Reservoir, Adv. Water Resour., 29: 1-10 (2006).
[25] Anbazhagan S., Thiruvengatam V., Kulanthai K., Adaptive Neuro-Fuzzy Inference System and Artificial Neural Network Modeling for the Adsorption of Methylene Blue by Novel Adsorbent in a Fixed - Bed Column Method, Iran. J. Chem. Chem. Eng. (IJCCE), 39: 75–93 (2020).
[26] Davoody M., Abdul Raman A.A., Asgharzadeh Ahmadi S., Binti Ibrahim S., Parthasarathy R., Determination of Volumetric Mass Transfer Coefficient in Gas-Solid-Liquid Stirred Vessels Handling High Solids Concentrations: Experiment and Modeling, Iran. J. Chem. Chem. Eng. (IJCCE), 37: 195–212 (2018).
[27] Kosorn W., Thavornyutikarn B., Janvikul W., Effects of Surface Treatments of Polycaprolactone Scaffolds on their Properties, Adv. Mat. Res., 747: 178-181 (2013).
[28] Uppanan P., Thavornyutikarn B., Kosorn W., Kaewkong P., Janvikul W., Enhancement of Chondrocyte Proliferation, Distribution, and Functions within Polycaprolactone Scaffolds by Surface Treatments, J. Biomed. Mater. Res. - Part A, 103: 2322-2332 (2015).
[29] Díaz E., Sandonis I., Valle M.B., In vitro Degradation of Poly (caprolactone)/nHA Composites, J. Nanomater., 2014: (2014).
[32] Qi H., Ye Z., Ren H., Chen N., Zeng Q., Wu X., Lu T., Bioactivity Assessment of PLLA/PCL/HAP Electrospun Nanofibrous Scaffolds for Bone Tissue Engineering, Life Sci., 148: 139-144 (2016).
[33] Zhou Z.H., He S.L., Huang T.L., Liu L.H., Liu Q.Q., Zhao Y.M., Ou B.L., Zeng W.N., Yang Z.M., Cao D.F., Degradation Behaviour and Biological Properties of Gelatin/hyaluronic Acid Composite Scaffolds, Mater. Res. Innov., 17: 420–424 (2013).
[34] Cummins K.A., Lee K.L., Cooper J.A., "Quantification of Entrapped Model Protein Released during Electrospun Nanofiber Degradation", 2015 41st Annual Northeast Biomedical Engineering Conference, NEBEC (2015).
[35] Zahabi A., Shahbazi E., Ahmadieh H., Hassani S.N., Totonchi M., Taei A., Masoudi N., Ebrahimi M., Aghdami N., Seifinejad A., Mehrnejad F., A New Efficient Protocol for Directed Differentiation of Retinal Pigmented Epithelial Cells from Normal and Retinal Disease Induced Pluripotent Stem Cells, Stem Cells Dev., 21: 2262-2272 (2012).
[36] Tezcaner A., Bugra K., Hasirci V., Retinal Pigment Epithelium Cell Culture on Surface Modified  Poly(hydroxybutyrate-co-hydroxyvalerate) Thin Films., Biomaterials, 24: 4573–4583 (2003).
[38] Thumann G., Viethen A., Gaebler A., Walter P., Kaempf S., Johnen S., Salz A.K., The In vitro and In vivo Behaviour of Retinal Pigment Epithelial Cells Cultured on Ultrathin Collagen Membranes, Biomaterials, 30: 287-294 (2009).
[39] Xiang P., Wu K.C., Zhu Y., Xiang L., Li C., Chen D.L., Chen F., Xu G., Wang A., Li M., Jin Z.B., A novel Bruch’s Membrane-mimetic Electrospun Substrate Scaffold for Human Retinal Pigment Epithelium Cells, Biomaterials, 35: 9777-9788 (2014).
[40] Park J.S., Kim J.M., Lee S.J., Lee S.G., Jeong Y.K., Kim S.E., Lee S.C., Surface Hydrolysis of Fibrous Poly(ε-caprolactone) Scaffolds for Enhanced Osteoblast Adhesion and Proliferation, Macromol. Res., 15: 424-429 (2007).
[41] McHugh K.J., Tao S.L., Saint-Geniez M., Porous Poly(ε-caprolactone) Scaffolds for Retinal Pigment Epithelium Transplantation, Investig. Ophthalmol. Vis. Sci., 55: 1754-1762 (2014).
[42] Guo C., Cai N., Dong Y., Duplex Surface Modification of Porous Poly (lactic acid) Scaffold, Mater. Lett., 94: 11–14 (2013).
[43] Abedalwafa M., Wang F., Wang L., Li C., Biodegradable Poly-epsilon-caprolactone (PCL) for Tissue Engineering Applications: A review, Rev. Adv. Mater. Sci., 34: 123-140 (2013).
[45] Gao J., Niklason L., Langer R., Surface Hydrolysis of Poly(glycolic acid) Meshes Increases the Seeding Density of Vascular Smooth Muscle Cells, J. Biomed. Mater. Res., 42: 417-424 (1998).
[46] Janvikul W., Uppanan P., Thavornyutikarn B., Kosorn W., Kaewkong P., Effects of Surface Topography, Hydrophilicity and Chemistry of Surface-treated PCL Scaffolds on Chondrocyte Infiltration and ECM Production, Procedia Eng., 59: 158-165 (2013).
[47] Williams R.L., Krishna Y., Dixon S., Haridas A., Grierson I., Sheridan C., Polyurethanes as Potential Substrates for Sub-retinal Retinal Pigment Epithelial  Cell Transplantation., J. Mater. Sci. Mater. Med., 16: 1087–1092 (2005).