Ocular Tissue Engineering Dimitrios Karamichos www.mdpi.com/journal/jfb Edited by Printed Edition of the Special Issue Published in Journal of Functional Biomaterials Journal of Functional Biomaterials Dimitrios Karamichos (Ed.) Ocular Tissue Engineering This book is a reprint of the Special Issue that appeared in the online, open access journal, Journal of Functional Biomaterials (ISSN 2079-4983) from 2015–2016 (available at: http://www.mdpi.com/journal/jfb/special_issues/ocular-tissue-eng). Guest Editor Dimitrios Karamichos University of Oklahoma Health Sciences Center, USA Editorial Office MDPI AG Klybeckstrasse 64 Basel, Switzerland Publisher Shu-Kun Lin Senior Assistant Editor Zhiqiao Dong 1. Edition 2016 MDPI • Basel • Beijing • Wuhan • Barcelona ISBN 978-3-03842-201-3 (Hbk) ISBN 978-3-03842-202-0 (PDF) Articles in this volume are Open Access and distributed under the Creative Commons Attribution license (CC BY), which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. The book taken as a whole is © 2016 MDPI, Basel, Switzerland, distributed under the terms and conditions of the Creative Commons by Attribution (CC BY-NC-ND) license (http://creativecommons.org/licenses/by-nc-nd/4.0/). III Table of Contents List of Contributors .................................................................................................... VII About the Guest Editor ............................................................................................... XI Preface to “Ocular Tissue Engineering” ................................................................... XIII Chapter 1: An Editorial Dimitrios Karamichos Ocular Tissue Engineering: Current and Future Directions Reprinted from: J. Funct. Biomater. 2015 , 6 (1), 77–80 http://www.mdpi.com/2079-4983/6/1/77 ...................................................................... 3 Chapter 2: Ocular Disease and Future Biomaterials Darren J. Lee Intraocular Implants for the Treatment of Autoimmune Uveitis Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 650–666 http://www.mdpi.com/2079-4983/6/3/650 .................................................................... 9 Cula N. Dautriche, Yangzi Tian, Yubing Xie and Susan T. Sharfstein A Closer Look at Schlemm’s Canal Cell Physiology: Implications for Biomimetics Reprinted from: J. Funct. Biomater. 2015, 6 (3), 963–985 http://www.mdpi.com/2079-4983/6/3/963 .................................................................. 28 Desiree' Lyon, Tina B. McKay, Akhee Sarkar-Nag, Shrestha Priyadarsini and Dimitrios Karamichos Human Keratoconus Cell Contractility is Mediated by Transforming Growth Factor-Beta Isoforms Reprinted from: J. Funct. Biomater. 2015 , 6 (2), 422–438 http://www.mdpi.com/2079-4983/6/2/422 .................................................................. 54 IV Audra M. A. Shadforth, Shuko Suzuki, Raphaelle Alzonne, Grant A. Edwards, Neil A. Richardson, Traian V. Chirila and Damien G. Harkin Incorporation of Human Recombinant Tropoelastin into Silk Fibroin Membranes with the View to Repairing Bruch’s Membrane Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 946–962 http://www.mdpi.com/2079-4983/6/3/946 .................................................................. 72 Jesintha Navaratnam, Tor P. Utheim, Vinagolu K. Rajasekhar and Aboulghassem Shahdadfar Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 917–945 http://www.mdpi.com/2079-4983/6/3/917 .................................................................. 90 Tor Paaske Utheim, Panagiotis Salvanos, Øygunn Aass Utheim, Sten Ræder, Lara Pasovic, Ole Kristoffer Olstad, Maria Fideliz de la Paz and Amer Sehic Transcriptome Analysis of Cultured Limbal Epithelial Cells on an Intact Amniotic Membrane following Hypothermic Storage in Optisol-GS Reprinted from: J. Funct. Biomater. 2016 , 7 (1), 4 http://www.mdpi.com/2079-4983/7/1/4 .....................................................................121 Shuko Suzuki, Rebecca A. Dawson, Traian V. Chirila, Audra M. A. Shadforth, Thomas A. Hogerheyde, Grant A. Edwards and Damien G. Harkin Treatment of Silk Fibroin with Poly(ethylene glycol) for the Enhancement of Corneal Epithelial Cell Growth Reprinted from: J. Funct. Biomater. 2015 , 6 (2), 345–366 http://www.mdpi.com/2079-4983/6/2/345 .................................................................136 Tor Paaske Utheim, Øygunn Aass Utheim, Qalb-E-Saleem Khan and Amer Sehic Culture of Oral Mucosal Epithelial Cells for the Purpose of Treating Limbal Stem Cell Deficiency Reprinted from: J. Funct. Biomater. 2016 , 7 (1), 5 http://www.mdpi.com/2079-4983/7/1/5 .....................................................................161 V Chapter 3: Ocular Nanotechnology and Tissue Engineering Yuhong Wang, Ammaji Rajala and Raju V. S. Rajala Lipid Nanoparticles for Ocular Gene Delivery Reprinted from: J. Funct. Biomater. 2015 , 6 (2), 379–394 http://www.mdpi.com/2079-4983/6/2/379 .................................................................187 Masatoshi Hirayama, Kazuo Tsubota and Takashi Tsuji Bioengineered Lacrimal Gland Organ Regeneration in Vivo Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 634–649 http://www.mdpi.com/2079-4983/6/3/634 .................................................................204 Martina Miotto, Ricardo M. Gouveia and Che J. Connon Peptide Amphiphiles in Corneal Tissue Engineering Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 687–707 http://www.mdpi.com/2079-4983/6/3/687 .................................................................221 Jon Roger Eidet, Darlene A. Dartt and Tor Paaske Utheim Concise Review: Comparison of Culture Membranes Used for Tissue Engineered Conjunctival Epithelial Equivalents Reprinted from: J. Funct. Biomater. 2015 , 6 (4), 1064–1084 http://www.mdpi.com/2079-4983/6/4/1064 ...............................................................245 Amer Sehic, Øygunn Aass Utheim, Kristoffer Ommundsen and Tor Paaske Utheim Pre-Clinical Cell-Based Therapy for Limbal Stem Cell Deficiency Reprinted from: J. Funct. Biomater. 2015 , 6 (3), 863–888 http://www.mdpi.com/2079-4983/6/3/863 .................................................................264 VII List of Contributors Raphaelle Alzonne Queensland Eye Institute, 140 Melbourne Street, South Brisbane, Queensland 4101, Australia. Traian V. Chirila Queensland Eye Institute, 140 Melbourne Street, South Brisbane, Queensland 4101, Australia; Australian Institute for Bioengineering and Nanotechnology, Faculty of Medicine and Biomedical Sciences, Science and Engineering Faculty, Queensland University of Technology, Brisbane, Queensland 4001, Australia; Faculty of Science, University of Western Australia, Crawley, Western Australia 6009, Australia. Che J. Connon Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle upon Tyne NE1 3BZ, UK. Darlene A. Dartt Schepens Eye Research Institute, Massachusetts Eye and Ear/Harvard Medical School, Boston, MA 02114, USA. Cula N. Dautriche State University of New York (SUNY) Polytechnic Institute, Colleges of Nanoscale Science and Engineering, 257 Fuller Road, Albany, NY 12203, USA. Rebecca A. Dawson Queensland Eye Institute, South Brisbane, Queensland 4101, Australia; School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001, Australia. Grant A. Edwards Australian Institute for Bioengineering and Nanotechnology, University of Queensland, St Lucia, Queensland 4072, Australia. Jon Roger Eidet Department of Ophthalmology, Oslo University Hospital, Oslo 0424, Norway. Maria Fideliz de la Paz El centro de Oftalmología Barraquer, Universitari Barraquer/Universitat Autonoma de Barcelona, Barcelona 08021, Spain. Ricardo M. Gouveia Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle upon Tyne NE1 3BZ, UK. Damien G. Harkin Queensland Eye Institute, South Brisbane, Queensland 4101, Australia; Institute of Health and Biomedical Innovation and School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001, Australia. Masatoshi Hirayama Department of Ophthalmology, Keio University School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan. VIII Thomas A. Hogerheyde Queensland Eye Institute, South Brisbane, Queensland 4101, Australia; Institute of Health and Biomedical Innovation, School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001, Australia. Dimitrios Karamichos Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Qalb-E-Saleem Khan Department of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø 9037, Norway. Darren J. Lee Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, 608 Stanton L. Young Blvd, DMEI PA404, Oklahoma City, OK 73104, USA. Desiree' Lyon Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Tina B. McKay Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Martina Miotto Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle upon Tyne NE1 3BZ, UK. Jesintha Navaratnam Department of Ophthalmology, Oslo University Hospital, Postbox 4950 Nydalen, Oslo 0424, Norway. Ole Kristoffer Olstad Department of Medical Biochemistry, Oslo University Hospital, Oslo 0407, Norway. Kristoffer Ommundsen Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, Oslo 0407, Norway. Lara Pasovic Department of Medical Biochemistry, Oslo University Hospital, Oslo 0407, Norway; Faculty of Medicine, University of Oslo, Oslo 0372, Norway. Shrestha Priyadarsini Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Sten Ræder The Norwegian Dry Eye Clinic, 0159 Oslo, Norway. Ammaji Rajala Dean A. McGee Eye Institute, Oklahoma City, OK 73104, USA; Department of Ophthalmology, College of Medicine, University of Oklahoma, Oklahoma City, OK 73014, USA. IX Raju V. S. Rajala Dean A. McGee Eye Institute, Oklahoma City, OK 73104, USA; Department of Ophthalmology, College of Medicine, University of Oklahoma, Oklahoma City, OK 73014, USA; Department of Physiology and Harold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73014, USA. Vinagolu K. Rajasekhar Memorial Sloan Kettering Cancer Center, Rockefeller Research Building, Room 1163, 430 East 67th Street/1275 York Avenue, New York, NY 10065, USA. Neil A. Richardson Queensland Eye Institute, 140 Melbourne Street, South Brisbane, Queensland 4101, Australia; School of Biomedical Sciences and Institute of Health & Biomedical Innovation, Queensland University of Technology, 2 George Street, Brisbane, Queensland 4001, Australia. Panagiotis Salvanos Department of Ophthalmology, Drammen Hospital, Vestre Viken Hospital Trust, Drammen 3004, Norway; Faculty of Medicine, University of Oslo, Oslo 0372, Norway. Akhee Sarkar-Nag Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. Amer Sehic Department of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, Oslo 0372, Norway; Department of Ophthalmology, Drammen Hospital, Vestre Viken Hospital Trust, Drammen 3004, Norway. Audra M. A. Shadforth Queensland Eye Institute, South Brisbane, Queensland 4101, Australia; School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001, Australia. Aboulghassem Shahdadfar Department of Ophthalmology, Oslo University Hospital, Postbox 4950 Nydalen, Oslo 0424, Norway. Susan T. Sharfstein State University of New York (SUNY) Polytechnic Institute, Colleges of Nanoscale Science and Engineering, 257 Fuller Road, Albany, NY 12203, USA. Shuko Suzuki Queensland Eye Institute, South Brisbane, Queensland 4101, Australia; Queensland Eye Institute, 140 Melbourne Street, South Brisbane, Queensland 4101, Australia. Yangzi Tian State University of New York (SUNY) Polytechnic Institute, Colleges of Nanoscale Science and Engineering, 257 Fuller Road, Albany, NY 12203, USA. Kazuo Tsubota Department of Ophthalmology, Keio University School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan. X Takashi Tsuji Laboratory of Organ Regeneration, RIKEN Center for Developmental Biology, Kobe, Hyogo 650-0047, Japan; Organ Technologies Inc., Chiyoda-ku, Tokyo 101-0048, Japan. Øygunn Aass Utheim Department of Oral Biology, Faculty of Dentistry, University of Oslo, Sognsvannsveien 10, Oslo 0372, Norway; Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, Oslo 0407, Norway. Tor Paaske Utheim Department of Medical Biochemistry, Oslo University Hospital, Oslo 0407, Norway; Department of Ophthalmology, Drammen Hospital, Vestre Viken Hospital Trust, Drammen 3004, Norway; Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo 0372, Norway; The Norwegian Dry Eye Clinic, 0159 Oslo, Norway; Faculty of Health Sciences, University College of South East Norway, Kongsberg 3603, Norway; Department of Ophthalmology, Drammen Hospital, Vestre Viken Hospital Trust, Drammen 3004, Norway. Yuhong Wang Dean A. McGee Eye Institute, Oklahoma City, OK 73104, USA; Department of Ophthalmology, College of Medicine, University of Oklahoma, Oklahoma City, OK 73014, USA. Yubing Xie State University of New York (SUNY) Polytechnic Institute, Colleges of Nanoscale Science and Engineering, 257 Fuller Road, Albany, NY 12203, USA. XI About the Guest Editor Dimitrios Karamichos is a corneal scientist and holds a Ph.D. in Tissue Engineering/Molecular Biology from the University of London, London, UK. He has dedicated his research career to wound healing and corneal diseases. Dr. Karamichos is currently an Assistant Professor of Ophthalmology and Cell Biology at the University of Oklahoma Health Sciences Center, Dean McGee Eye Institute, Oklahoma, USA. He is also the Vice President of the Graduate College of the University of Oklahoma Health Sciences Center and the President of the Society for Neuroscience, Oklahoma Chapter. Dr. Karamichos's research interests include corneal wound healing and fibrosis, keratoconus, corneal diabetes, and drug delivery. He is the principal investigator on numerous grants funded by the National Eye Institute/National Institute of Health. XIII Preface to “Ocular Tissue Engineering” Tissue engineering is a rapidly growing area, and complex three-dimensional tissue substitutes are emerging. Although cells are routinely cultured outside the body, current research shows that tissue engineered constructs can be used as replacement tissues for damaged or diseased human organs. This book is an outgrowth of a Special Issue of the Journal of Functional Biomaterials (JFB) devoted to Ocular Tissue Engineering and contains both original research and review articles. Each of the articles included here provides an up-to-date analysis and cutting edge technology in this fast growing field. Biomaterials and nanotechnology in cellular processes, as well as in ocular disease, are highlighted. We sincerely hope that readers will enjoy these articles and be inspired by the ideas presented. Dimitrios Karamichos Guest Editor Chapter 1: An Editorial Ocular Tissue Engineering: Current and Future Directions D. Karamichos Reprinted from J. Funct. Biomater. Cite as: Karamichos, D. Ocular Tissue Engineering: Current and Future Directions. J. Funct. Biomater. 2015 , 6 , 77–80. Tissue engineering (TE) is a concept that was first emerged in the early 1990s to provide solutions to severe injured tissues and/or organs [ 1 ]. The dream was to be able to restore and replace the damaged tissue with an engineered version which would ultimately help overcome problems such as donor shortages, graft rejections, and inflammatory responses following transplantation. While an incredible amount of progress has been made, suggesting that TE concept is viable, we are still not able to overcome major obstacles. In TE, there are two main strategies that researchers have adopted: (1) cell-based, where cells are been manipulated to create their own environment before transplanted to the host, and (2) scaffold-based, where an extracellular matrix is created to mimic in vivo structures. TE approaches for ocular tissues are available and have indeed come a long way, over the last decades; however more clinically relevant ocular tissue substitutes are needed. Figure 1 highlights the importance of TE in ocular applications and indicates the avenues available based on each tissue. In cornea, TE approaches are vital in order to maintain the transparent barrier between the eye and the environment. Of the three corneal layers (epithelium, stroma, and endothelium) probably the most difficult one to replace is the stroma. Stroma is a thick, transparent middle layer, consisting of regularly arranged collagen fibers along with sparsely distributed resident cells commonly known as keratocytes. The corneal stroma consists of approximately 200 collagen fibril layers and account for up to 90% of the total corneal thickness. Corneal transplantation is currently the only surgical procedure for replacing damaged or diseased corneas. Damaged cornea is replaced by donated corneal tissue in its entirety (penetrating keratoplasty) or in part (lamellar keratoplasty). While the surgical procedure has been somewhat successful, major problems remain including donor corneas shortage, risks of infection, and graft rejection. In an attempt for an alternative avenue, several studies have reported successful cultivation of corneal stroma, in combination with corneal epithelium and endothelium, however the long-term in vivo data and clinical applications are still lacking [ 1 ]. The corneal epithelium has been targeted by scientists and a variety of TE applications using both cell and scaffold-based approaches have been developed [ 2 – 6 ]. Studies reporting the successful transplantation of mucosal epithelial cells [ 5 , 6 ] as well as limbal stem cells [ 2 ] are promising. Tissue grafts 3 such amniotic membranes [ 3 , 4 ] have also been reported and used in humans. While these have been assessed in clinical setting, long-term studies are still needed in order to safely assess the benefits. In lens, despite the limited number of studies developing TE solutions, there is a clear need for cataract surgeries alternatives. Currently, lens opacification or else known as cataracts are treated surgically by removing the lens and replacing it with artificial intraocular lenses (IOL) [ 1 , 7 ]. Most of the people receiving cataract surgery will need to come back for a second surgery due to the posterior capsule opacification (PCO). PCO occurs because lens epithelial cells remaining after cataract surgery have grown on the capsule causing it to become hazy and opaque [ 1 , 7 , 8 ]. Development of alternatives is almost nonexistent and urgently needed. One of the few TE approaches was reported by Tsionis et al. [ 9 ] where a human retinal PE cell line cultured in Matrigel was differentiated in lentoids and lens-like structures. Nevertheless, therapies based on this technique or others are far away and it remains unknown if TE is the future for lens related clinical problems. In retina, both cell and substrate-based TE approaches have been reported mainly in animal models. Homologous retinal pigment epithelium (RPE) cells have been transplanted in the subretinal space with no visual benefits to the patients [ 10 , 11 ]. On the other hand autologous RPE transplantation resulted in clinically significant improvement of vision; however the limited number of healthy cells that can be isolated from the patient is a huge problem [ 12 , 13 ]. The concept of the use of polymers for retinal TE is rather new and has only been emerged in the last decade or so. As reviewed by Trese and co-authors [ 14 ] the ideal polymer for retinal transplantation should be thinner than 50 μ m, porous, biodegradable, and have the correct Young’s modulus. Several polymers fulfill this criteria including but not limited to poly(lactic-co-glycolic acid) (PLGA), poly(lactic acid (PLLA), poly(glucerol-sebacate) (PGS), and poly(caprolactone) (PCL) [ 14 , 15 ]. However, only a few studies have shown promising results using these or other polymers for TE retinal applications. The combination of PLLA-PLGA polymer reported by Thomson and co-authors [ 16 ] showed good RPE cellular viability, adhesion and proliferation for the course of the month long study. However, the main limitation of this study was the use of cell lines instead of primary cells which are known to be different in terms of their behavior. The general consensus is that embryonic stem cells (ESC) and induced pluripotent stem (iPS) cells are a better choice since they more closely resemble actual RPE. This, however, remains to be seen. Regardless of the cell source, technical challenges still remain before cell-substrate based therapies can be successful. 4 as well as limbal stem cells [2] are prom ising. Tissue grafts such amniotic membranes [3,4 been reported and used in humans. While these have been assessed in clinical setting still needed in order to safely assess the benefits. Figure 1. Schematic diagram highlighting the importance of tissue engineering (TE) approaches in ocular tissues: cornea, lens, and retina. In lens, despite the limited number of studies developing TE solutions, there is a clear need fo alternatives. Cu rrently, lens opacification or else known as cataracts are treate removing the lens and replacing it with artificial intraocular lenses (IOL) [1,7]. Most o receiving cataract surger y will need to come back for a second surgery due to the posterio (PCO). PCO occurs because lens epithelial cells remaining after cataract surger on the capsule causing it to become hazy and opaque [1,7,8]. Development of alternatives urgently needed. One of the few TE approaches was reported by Tsionis et al. [9 human retinal PE cell line cultured in Matrigel was differentiated in lentoids and lens-like based on this technique or others are far away and it remains unknow the future for lens related clinical problems. In retina, both cell and substrate-based TE approaches have been reported mainly in animal model epithelium (RPE) cells have been transplanted in the subretinal space wit benefits to the patients [10,11]. On the other hand autologous RPE transplantation resulted i improvement of vision; however the limited number of healthy cells that can b patient is a huge problem [12,13]. The concept of the use of polymers for retinal TE has only been emerged in the last decade or so. As reviewed by Trese and co-authors [14 polymer for retinal transplantation should be thinner than 50 μ m, porous, biodegradable, an Tissue repair/Tissue regeneration Tissue repair/Tissue regeneration Tissue Engineering Lens Retina Cornea Scaffold ‐ based Cell ‐ based Figure 1. Schematic diagram highlighting the importance of tissue engineering (TE) approaches in ocular tissues: cornea, lens, and retina. In conclusion, the human eye with the different structures, cell types, and tissues is an ideal candidate for TE approaches. The eye structures and the inadequate to-date therapies make this a very attractive tissue for TE. This is well understood within the scientific community and that is why significant discoveries and knowledge advancements have been made. Perhaps the one tissue with the most success is the corneal epithelium. There is no reason why the other structures cannot be regenerated or reconstructed using TE techniques. The challenge here is to be able to get the scientists, engineers, and clinicians to work together in order to tackle today’s challenges and give our patients the best possible treatment. Conflicts of Interest: The author declares no conflict of interest. Acknowledgments: The author would like to thank the National Eye Institute of the National Institutes of Health for the support (EY023568 and EY020886). The content is solely the responsibility of the author and does not necessarily represent the official views of the National Institutes of Health. 5