{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T02:22:00Z","timestamp":1789179720347,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,3,5]],"date-time":"2024-03-05T00:00:00Z","timestamp":1709596800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Agence de la Biom\u00e9decine"},{"name":"Appel d\u2019Offre Recherche et Greffe 2022"},{"name":"Fondation des Gueules Cass\u00e9es"},{"name":"Union des Bless\u00e9s de la Face et de la T\u00eate (UBFT)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>The bioengineering of corneal endothelial grafts consists of seeding in vitro cultured corneal endothelial cells onto a thin, transparent, biocompatible, and sufficiently robust carrier which can withstand surgical manipulations. This is one of the most realistic alternatives to donor corneas, which are in chronic global shortage. The anterior capsule of the crystalline lens has already been identified as one of the best possible carriers, but its challenging manual preparation has limited its use. In this study, we describe a femtosecond laser cutting process of the anterior capsule of whole lenses in order to obtain capsule discs of 8 mm diameter, similar to conventional endothelial grafts. Circular marks made on the periphery of the disc indicate its orientation. Immersion in water for 3 days is sufficient to completely remove the lens epithelial cells and to enable the seeding of corneal endothelial cells, which remain viable after 27 days of culture. Therefore, this method provides a transparent, decellularized disc ready to form viable tissue engineered endothelial grafts.<\/jats:p>","DOI":"10.3390\/bioengineering11030255","type":"journal-article","created":{"date-parts":[[2024,3,5]],"date-time":"2024-03-05T11:26:59Z","timestamp":1709638019000},"page":"255","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Femtosecond Laser Cutting of Human Crystalline Lens Capsule and Decellularization for Corneal Endothelial Bioengineering"],"prefix":"10.3390","volume":"11","author":[{"given":"Olfa","family":"Ben Moussa","sequence":"first","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2601-6350","authenticated-orcid":false,"given":"Louise","family":"Parveau","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"In\u00e8s","family":"Aouimeur","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gr\u00e9gory","family":"Egaud","sequence":"additional","affiliation":[{"name":"GIE Manutech-USD, 18 Rue Professeur Beno\u00eet Lauras, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0080-3423","authenticated-orcid":false,"given":"Corantin","family":"Maurin","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofiane","family":"Fraine","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00e9bastien","family":"Urbaniak","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chantal","family":"Perrache","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiguo","family":"He","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4072-492X","authenticated-orcid":false,"given":"Sedao","family":"Xxx","sequence":"additional","affiliation":[{"name":"Laboratoire Hubert Curien, Jean Monnet University, 18 Rue Professeur Beno\u00eet Lauras, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oliver","family":"Dorado Cortez","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"},{"name":"Ophthalmology Department, University Hospital, Avenue Albert Raimond, CEDEX 02, 42055 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3660-6541","authenticated-orcid":false,"given":"Sylvain","family":"Poinard","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"},{"name":"Ophthalmology Department, University Hospital, Avenue Albert Raimond, CEDEX 02, 42055 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5723-2281","authenticated-orcid":false,"given":"Cyril","family":"Mauclair","sequence":"additional","affiliation":[{"name":"Laboratoire Hubert Curien, Jean Monnet University, 18 Rue Professeur Beno\u00eet Lauras, 42000 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philippe","family":"Gain","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"},{"name":"Ophthalmology Department, University Hospital, Avenue Albert Raimond, CEDEX 02, 42055 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3486-2875","authenticated-orcid":false,"given":"Gilles","family":"Thuret","sequence":"additional","affiliation":[{"name":"Laboratory Biology, Engineering and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health Innovation Campus, Jean Monnet University, 10 Rue de la Marandi\u00e8re, 42270 Saint-Priest-en-Jarez, France"},{"name":"Ophthalmology Department, University Hospital, Avenue Albert Raimond, CEDEX 02, 42055 Saint-Etienne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"29047","DOI":"10.1038\/srep29047","article-title":"3D map of the human corneal endothelial cell","volume":"6","author":"He","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_2","first-page":"987","article-title":"Descemet membrane endothelial keratoplasty (DMEK)","volume":"25","author":"Melles","year":"2006","journal-title":"Cornea"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2171","DOI":"10.1016\/j.ophtha.2006.06.034","article-title":"Trends of Penetrating Keratoplasty in the United States from 1980 to 2004","volume":"113","author":"Darlington","year":"2006","journal-title":"Ophthalmology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"108462","DOI":"10.1016\/j.exer.2021.108462","article-title":"Focus on cell therapy to treat corneal endothelial diseases","volume":"204","author":"Faye","year":"2021","journal-title":"Exp. Eye Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1097\/j.jcrs.0000000000000520","article-title":"Outcomes of corneal transplantation in Europe: Report by the European Cornea and Cell Transplantation Registry","volume":"47","author":"Dunker","year":"2021","journal-title":"J. Cataract. Refract. Surg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e0008","DOI":"10.1097\/ebct.0000000000000008","article-title":"2022 Eye Banking Statistical Report\u2014Executive Summary","volume":"2","author":"Mathews","year":"2023","journal-title":"Eye Bank. Corneal Transplant."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1001\/jamaophthalmol.2015.4776","article-title":"Global Survey of Corneal Transplantation and Eye Banking","volume":"134","author":"Gain","year":"2016","journal-title":"JAMA Ophthalmol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"100987","DOI":"10.1016\/j.preteyeres.2021.100987","article-title":"Approaches for corneal endothelium regenerative medicine","volume":"87","author":"Thuret","year":"2022","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1056\/NEJMoa1712770","article-title":"Injection of Cultured Cells with a ROCK Inhibitor for Bullous Keratopathy","volume":"378","author":"Kinoshita","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_10","unstructured":"Thuret, G., Muraine, M., and Gain, P. (2020). The Cornea Endothelium. Issy-les Moulineaux, Elsevier."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"6087","DOI":"10.1038\/s41598-019-42493-3","article-title":"Functional Evaluation of Two Corneal Endothelial Cell-Based Therapies: Tissue-Engineered Construct and Cell Injection","volume":"9","author":"Peh","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"14149","DOI":"10.1038\/s41598-017-14723-z","article-title":"Regulatory Compliant Tissue-Engineered Human Corneal Endothelial Grafts Restore Corneal Function of Rabbits with Bullous Keratopathy","volume":"7","author":"Peh","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"7955","DOI":"10.1038\/srep07955","article-title":"In Vitro Expansion of Corneal Endothelial Cells on Biomimetic Substrates","volume":"5","author":"Palchesko","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1007\/s00586-008-0745-3","article-title":"Scaffolding in tissue engineering: General approaches and tissue-specific considerations","volume":"17","author":"Chan","year":"2008","journal-title":"Eur. Spine J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"800","DOI":"10.1167\/iovs.03-0016","article-title":"Amniotic Membrane as a Carrier for Cultivated Human Corneal Endothelial Cell Transplantation","volume":"45","author":"Ishino","year":"2004","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"917","DOI":"10.3390\/jfb6030917","article-title":"Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium","volume":"6","author":"Navaratnam","year":"2015","journal-title":"J. Funct. Biomater."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1001\/archophthalmol.2010.372","article-title":"Prevalence of and Risk Factors for Cornea Guttata in a Population-Based Study in a Southwestern Island of Japan: The Kumejima Study","volume":"129","author":"Higa","year":"2011","journal-title":"Arch. Ophthalmol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Arnalich-Montiel, F., Moratilla, A., Fuentes-Juli\u00e1n, S., Aparicio, V., Cadenas Martin, M., Peh, G., Mehta, J.S., Adnan, K., Porrua, L., and P\u00e9rez-Sarriegui, A. (2019). Treatment of corneal endothelial damage in a rabbit model with a bioengineered graft using human decellularized corneal lamina and cultured human corneal endothelium. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0225480"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1097\/ICO.0b013e31818c2c36","article-title":"Human Anterior Lens Capsule as Carrier Matrix for Cultivated Human Corneal Endothelial Cells","volume":"28","author":"Yoeruek","year":"2009","journal-title":"Cornea"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.exer.2008.08.002","article-title":"The lens capsule","volume":"88","author":"Danysh","year":"2009","journal-title":"Exp. Eye Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e2020","DOI":"10.1002\/term.2633","article-title":"Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering","volume":"12","author":"Zakaria","year":"2018","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1111\/aos.15081","article-title":"Tissue engineered endothelial keratoplasty in rabbit: Tips and tricks","volume":"100","author":"Crouzet","year":"2022","journal-title":"Acta Ophthalmol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"922","DOI":"10.1016\/j.ajo.2010.01.005","article-title":"A Method to Confirm Correct Orientation of Descemet Membrane During Descemet Membrane Endothelial Keratoplasty","volume":"149","author":"Bachmann","year":"2010","journal-title":"Am. J. Ophthalmol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6639","DOI":"10.1167\/iovs.16-20256","article-title":"Cutting and Decellularization of Multiple Corneal Stromal Lamellae for the Bioengineering of Endothelial Grafts","volume":"57","author":"He","year":"2016","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Villamil Ballesteros, A.C., Segura Puello, H.R., Lopez-Garcia, J.A., Bernal-Ballen, A., Nieto Mosquera, D.L., Mu\u00f1oz Forero, D.M., Segura Charry, J.S., and Neira Bejarano, Y.A. (2020). Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin. Polymers, 12.","DOI":"10.3390\/polym12030590"},{"key":"ref_26","unstructured":"(2024, January 18). Luna\u00ae Universal Probe qPCR Master Mix|NEB. Available online: https:\/\/www.neb.com\/en\/products\/m3004-luna-universal-probe-qpcr-master-mix#Protocols,%20Manuals%20&%20Usage_Manuals."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1007\/s10561-021-09918-8","article-title":"Corneal endothelial cell therapy: Feasibility of cell culture from corneas stored in organ culture","volume":"22","author":"He","year":"2021","journal-title":"Cell Tissue Bank."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6018","DOI":"10.1167\/iovs.10-6641","article-title":"Pan-Corneal Endothelial Viability Assessment: Application to Endothelial Grafts Predissected by Eye Banks","volume":"52","author":"Pipparelli","year":"2011","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_29","first-page":"1345","article-title":"Optimization of immunostaining on flat-mounted human corneas","volume":"21","author":"Forest","year":"2015","journal-title":"Mol. Vis."},{"key":"ref_30","unstructured":"(2023, December 19). Challenges in Corneal Endothelial Cell Culture. Available online: https:\/\/www.futuremedicine.com\/doi\/epub\/10.2217\/rme-2020-0202."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Aouimeur, I., Sagnial, T., Coulomb, L., Maurin, C., Thomas, J., Forestier, P., Ninotta, S., Perrache, C., Forest, F., and Gain, P. (2023). Investigating the Role of TGF-\u03b2 Signaling Pathways in Human Corneal Endothelial Cell Primary Culture. Cells, 12.","DOI":"10.3390\/cells12121624"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1007\/s10561-013-9414-9","article-title":"Non-invasive measurement of transparency, arcus senilis, and scleral rim diameter of corneas during eye banking","volume":"15","author":"Acquart","year":"2014","journal-title":"Cell Tissue Bank."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Weigert, M., Schmidt, U., Haase, R., Sugawara, K., and Myers, G. (2020, January 5). Star-convex Polyhedra for 3D Object Detection and Segmentation in Microscopy. Proceedings of the 2020 IEEE Winter Conference on Applications of Computer Vision (WACV), Snowmass Village, CO, USA.","DOI":"10.1109\/WACV45572.2020.9093435"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Frangi, A.F., Schnabel, J.A., Davatzikos, C., Alberola-L\u00f3pez, C., and Fichtinger, G. (2018). Proceedings of the Medical Image Computing and Computer Assisted Intervention\u2014MICCAI 2018, Springer International Publishing.","DOI":"10.1007\/978-3-030-00931-1"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1016\/j.ophtha.2020.09.002","article-title":"Five-Year Follow-up of First 11 Patients Undergoing Injection of Cultured Corneal Endothelial Cells for Corneal Endothelial Failure","volume":"128","author":"Numa","year":"2021","journal-title":"Ophthalmology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1097\/ICO.0000000000001578","article-title":"Outcomes of Hemi-Descemet Membrane Endothelial Keratoplasty for Fuchs Endothelial Corneal Dystrophy","volume":"37","author":"Birbal","year":"2018","journal-title":"Cornea"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1097\/ICO.0000000000002127","article-title":"Quarter-Descemet Membrane Endothelial Keratoplasty: One- to Two-Year Clinical Outcomes","volume":"39","author":"Birbal","year":"2020","journal-title":"Cornea"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1167\/iovs.61.4.11","article-title":"Age-Related Changes to the Three-Dimensional Full Shape of the Isolated Human Crystalline Lens","volume":"61","author":"Mohamed","year":"2020","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1146\/annurev-vision-102016-061400","article-title":"EK (DLEK, DSEK, DMEK): New Frontier in Cornea Surgery","volume":"3","author":"Price","year":"2017","journal-title":"Annu. Rev. Vis. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Garcin, T., Gain, P., and Thuret, G. (Eye, 2022). Femtosecond laser-cut autologous anterior lens capsule transplantation to treat refractory macular holes, Eye, Epub ahead of print.","DOI":"10.1038\/s41433-022-02062-x"}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/11\/3\/255\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:09:39Z","timestamp":1760105379000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/11\/3\/255"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,5]]},"references-count":40,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2024,3]]}},"alternative-id":["bioengineering11030255"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering11030255","relation":{},"ISSN":["2306-5354"],"issn-type":[{"value":"2306-5354","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,5]]}}}