T H van Essen1, L van Zijl2, T Possemiers3, A A Mulder4, S J Zwart5, C-H Chou6, C C Lin7, H J Lai8, G P M Luyten9, M J Tassignon10, N Zakaria11, A El Ghalbzouri12, M J Jager13. 1. Department of Ophthalmology, J3-S, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Electronic address: T.H.van_Essen@lumc.nl. 2. Department of Research, Aeon Astron Europe B.V., J.H. Oortweg 19, 2333 CH, Leiden, The Netherlands. Electronic address: Lisanne.van.zijl@aeonastron.com. 3. Department of Ophthalmology, Antwerp University Hospital, Wilrijkstraat 10, 2650 Edegem, Belgium. Electronic address: Tine.Possemiers@hotmail.be. 4. Department of Molecular Cell-biology, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Electronic address: A.A.Mulder@lumc.nl. 5. Department of Research, Aeon Astron Europe B.V., J.H. Oortweg 19, 2333 CH, Leiden, The Netherlands. Electronic address: Sarah.zwart@aeonastron.com. 6. Department of Research, Body Organ Biomedical Corporation, 5F, No. 153, Section 3, Xinyi Road, Da'an District, Taipei City 106, Taiwan, ROC. Electronic address: Charles.chou@aeonastron.com. 7. Department of Research, Body Organ Biomedical Corporation, 5F, No. 153, Section 3, Xinyi Road, Da'an District, Taipei City 106, Taiwan, ROC. Electronic address: Julio.Lin@aeonastron.com. 8. Department of Research, Aeon Astron Europe B.V., J.H. Oortweg 19, 2333 CH, Leiden, The Netherlands. Electronic address: Michael.Lai@aeonastron.com. 9. Department of Ophthalmology, J3-S, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Electronic address: G.P.M.Luyten@lumc.nl. 10. Department of Ophthalmology, Antwerp University Hospital, Wilrijkstraat 10, 2650 Edegem, Belgium. Electronic address: Marie-Jose.Tassignon@uza.be. 11. Department of Ophthalmology, Antwerp University Hospital, Wilrijkstraat 10, 2650 Edegem, Belgium; University of Antwerp, Prinsstraat 13, 2000 Antwerpen, Belgium. Electronic address: Nadia.zakaria@gmail.com. 12. Department of Molecular Cell-biology, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Electronic address: A.E.L.Ghalbzouri@lumc.nl. 13. Department of Ophthalmology, J3-S, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands. Electronic address: M.J.Jager@lumc.nl.
Abstract
PURPOSE: To determine whether a fish scale-derived collagen matrix (FSCM) meets the basic criteria to serve as an artificial cornea, as determined with in vitro and in vivo tests. METHODS: Primary corneal epithelial and stromal cells were obtained from human donor corneas and used to examine the (in)direct cytotoxicity effects of the scaffold. Cytotoxicity was assessed by an MTT assay, while cellular proliferation, corneal cell phenotype and adhesion markers were assessed using an EdU-assay and immunofluorescence. For in vivo-testing, FSCMs were implanted subcutaneously in rats. Ologen(®) Collagen Matrices were used as controls. A second implant was implanted as an immunological challenge. The FSCM was implanted in a corneal pocket of seven New Zealand White rabbits, and compared to sham surgery. RESULTS: The FSCM was used as a scaffold to grow corneal epithelial and stromal cells, and displayed no cytotoxicity to these cells. Corneal epithelial cells displayed their normal phenotypical markers (CK3/12 and E-cadherin), as well as cell-matrix adhesion molecules: integrin-α6 and β4, laminin 332, and hemi-desmosomes. Corneal stromal cells similarly expressed adhesion molecules (integrin-α6 and β1). A subcutaneous implant of the FSCM in rats did not induce inflammation or sensitization; the response was comparable to the response against the Ologen(®) Collagen Matrix. Implantation of the FSCM in a corneal stromal pocket in rabbits led to a transparent cornea, healthy epithelium, and, on histology, hardly any infiltrating immune cells. CONCLUSION: The FSCM allows excellent cell growth, is not immunogenic and is well-tolerated in the cornea, and thus meets the basic criteria to serve as a scaffold to reconstitute the cornea.
PURPOSE: To determine whether a fish scale-derived collagen matrix (FSCM) meets the basic criteria to serve as an artificial cornea, as determined with in vitro and in vivo tests. METHODS: Primary corneal epithelial and stromal cells were obtained from humandonor corneas and used to examine the (in)direct cytotoxicity effects of the scaffold. Cytotoxicity was assessed by an MTT assay, while cellular proliferation, corneal cell phenotype and adhesion markers were assessed using an EdU-assay and immunofluorescence. For in vivo-testing, FSCMs were implanted subcutaneously in rats. Ologen(®) Collagen Matrices were used as controls. A second implant was implanted as an immunological challenge. The FSCM was implanted in a corneal pocket of seven New Zealand White rabbits, and compared to sham surgery. RESULTS: The FSCM was used as a scaffold to grow corneal epithelial and stromal cells, and displayed no cytotoxicity to these cells. Corneal epithelial cells displayed their normal phenotypical markers (CK3/12 and E-cadherin), as well as cell-matrix adhesion molecules: integrin-α6 and β4, laminin 332, and hemi-desmosomes. Corneal stromal cells similarly expressed adhesion molecules (integrin-α6 and β1). A subcutaneous implant of the FSCM in rats did not induce inflammation or sensitization; the response was comparable to the response against the Ologen(®) Collagen Matrix. Implantation of the FSCM in a corneal stromal pocket in rabbits led to a transparent cornea, healthy epithelium, and, on histology, hardly any infiltrating immune cells. CONCLUSION: The FSCM allows excellent cell growth, is not immunogenic and is well-tolerated in the cornea, and thus meets the basic criteria to serve as a scaffold to reconstitute the cornea.
Authors: Lingjia Liu; Wenbo Cheng; Di Wu; Luxia Chen; Shasha Yu; Tong Zuo; Lin Zhang; Kun Yang; Hua Li; Hui Zhang; Pinghui Wei; Alex Lap Ki Ng; George Pak-Man Cheng; Victor Chi-Pang Woo; Jia Yin; Kin Chiu; Yan Wang Journal: Invest Ophthalmol Vis Sci Date: 2020-05-11 Impact factor: 4.799
Authors: Susanne G Pondorfer; Maximilian W M Wintergerst; Shekoufeh Gorgi Zadeh; Thomas Schultz; Manuel Heinemann; Frank G Holz; Robert P Finger Journal: Invest Ophthalmol Vis Sci Date: 2020-03-09 Impact factor: 4.799