Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity
Sterilization of biodegradable, collagen-based implants is challenging as irradiation sterilization methods can alter their mechanical properties. Electron beam (EB) irradiation is a terminal sterilization method that has been used for biologically-derived implants. Here, recombinant human collagen...
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.883977/full |
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| author | Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Mohammed Mirazul Islam Mohammed Mirazul Islam Oleksiy Buznyk Oleksiy Buznyk Elle Edin Elle Edin Elle Edin Elle Edin Elle Edin Marc Groleau Marc Groleau Monika Kozak-Ljunggren Federica M. Magrelli Dina B. AbuSamra Pablo Argüeso James Chodosh Aneta Liszka Per Fagerholm May Griffith May Griffith May Griffith May Griffith |
| author_facet | Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Mohammed Mirazul Islam Mohammed Mirazul Islam Oleksiy Buznyk Oleksiy Buznyk Elle Edin Elle Edin Elle Edin Elle Edin Elle Edin Marc Groleau Marc Groleau Monika Kozak-Ljunggren Federica M. Magrelli Dina B. AbuSamra Pablo Argüeso James Chodosh Aneta Liszka Per Fagerholm May Griffith May Griffith May Griffith May Griffith |
| author_sort | Fiona C. Simpson |
| collection | DOAJ |
| description | Sterilization of biodegradable, collagen-based implants is challenging as irradiation sterilization methods can alter their mechanical properties. Electron beam (EB) irradiation is a terminal sterilization method that has been used for biologically-derived implants. Here, recombinant human collagen type III-phosphorylcholine (RHCIII-MPC) hydrogels were irradiated with EB doses of 17, 19, or 21 kGy and their subsequent biocompatibility and ability to promote regeneration in rabbit corneas was evaluated. Unirradiated hydrogels stored in 1% chloroform in phosphate-buffered saline (C-PBS) were the controls. There were no significant differences between irradiated and non-irradiated samples in optical or physical properties (tensile strength, modulus, elasticity), or the ability to support cell growth. However, irradiated implants were more sensitive to high levels of collagenase than unirradiated controls and the C-PBS implants had increased cell growth compared to EB and controls at 72 h. Corneal implants e-beamed at 17 kGy or e-beamed and subsequently frozen (EB-F) to increase shelf-life showed no adverse biological effects of the irradiation. EB, EB-F, and C-PBS implanted corneas all rapidly re-epithelialized but showed mild neovascularization that resolved over 6 months. The regenerated neo-corneas were transparent at 6 months post-operation. In vivo confocal microscopy confirmed normal morphology for the epithelium, stroma, sub-basal nerves and unoperated endothelium. Histology showed that all the regenerated corneas were morphologically similar to the normal. Immunohistochemistry indicated the presence of a differentiated corneal epithelium and functional tear film. In conclusion, the e-beamed corneal implants performed as well as non-irradiated control implants, resulting in fully regenerated neo-corneas with new nerves and without blood vessels or inflammation that may impede vision or corneal function. Therefore, a complete validation study to establish EB irradiation as an effective means for corneal implant sterilization prior to clinical application is necessary as a next step. |
| first_indexed | 2024-12-12T05:41:52Z |
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| last_indexed | 2024-12-12T05:41:52Z |
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| spelling | doaj.art-cb497156d0fa40958596512b9786d7e02022-12-22T00:35:53ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-06-011010.3389/fbioe.2022.883977883977Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration CapacityFiona C. Simpson0Fiona C. Simpson1Fiona C. Simpson2Fiona C. Simpson3Mohammed Mirazul Islam4Mohammed Mirazul Islam5Oleksiy Buznyk6Oleksiy Buznyk7Elle Edin8Elle Edin9Elle Edin10Elle Edin11Elle Edin12Marc Groleau13Marc Groleau14Monika Kozak-Ljunggren15Federica M. Magrelli16Dina B. AbuSamra17Pablo Argüeso18James Chodosh19Aneta Liszka20Per Fagerholm21May Griffith22May Griffith23May Griffith24May Griffith25Department of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenMaisonneuve-Rosemont Hospital Research Centre, Montréal, QC, CanadaDepartment of Ophthalmology, Institute of Biomedical Engineering, Université de Montréal, Montréal, QC, CanadaCentre de Recherche—Centre Hospitalier de l’Université de Montréal, Montréal, QC, CanadaDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenDepartment of Ophthalmology, Massachusetts Eye and Ear and Schepens Eye Research Institute, Harvard Medical School, Boston, MA, United StatesDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenFilatov Institute of Eye Diseases and Tissue Therapy of the NAMS of Ukraine, Odessa, UkraineDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenMaisonneuve-Rosemont Hospital Research Centre, Montréal, QC, CanadaDepartment of Ophthalmology, Institute of Biomedical Engineering, Université de Montréal, Montréal, QC, CanadaCentre de Recherche—Centre Hospitalier de l’Université de Montréal, Montréal, QC, CanadaDepartment of Polymer Chemistry, Uppsala University, Uppsala, SwedenMaisonneuve-Rosemont Hospital Research Centre, Montréal, QC, CanadaCentre de Recherche—Centre Hospitalier de l’Université de Montréal, Montréal, QC, CanadaDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenInterdepartmental Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, ItalyDepartment of Ophthalmology, Massachusetts Eye and Ear and Schepens Eye Research Institute, Harvard Medical School, Boston, MA, United StatesDepartment of Ophthalmology, Massachusetts Eye and Ear and Schepens Eye Research Institute, Harvard Medical School, Boston, MA, United StatesDepartment of Ophthalmology, Massachusetts Eye and Ear and Schepens Eye Research Institute, Harvard Medical School, Boston, MA, United StatesDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, SwedenMaisonneuve-Rosemont Hospital Research Centre, Montréal, QC, CanadaDepartment of Ophthalmology, Institute of Biomedical Engineering, Université de Montréal, Montréal, QC, CanadaCentre de Recherche—Centre Hospitalier de l’Université de Montréal, Montréal, QC, CanadaSterilization of biodegradable, collagen-based implants is challenging as irradiation sterilization methods can alter their mechanical properties. Electron beam (EB) irradiation is a terminal sterilization method that has been used for biologically-derived implants. Here, recombinant human collagen type III-phosphorylcholine (RHCIII-MPC) hydrogels were irradiated with EB doses of 17, 19, or 21 kGy and their subsequent biocompatibility and ability to promote regeneration in rabbit corneas was evaluated. Unirradiated hydrogels stored in 1% chloroform in phosphate-buffered saline (C-PBS) were the controls. There were no significant differences between irradiated and non-irradiated samples in optical or physical properties (tensile strength, modulus, elasticity), or the ability to support cell growth. However, irradiated implants were more sensitive to high levels of collagenase than unirradiated controls and the C-PBS implants had increased cell growth compared to EB and controls at 72 h. Corneal implants e-beamed at 17 kGy or e-beamed and subsequently frozen (EB-F) to increase shelf-life showed no adverse biological effects of the irradiation. EB, EB-F, and C-PBS implanted corneas all rapidly re-epithelialized but showed mild neovascularization that resolved over 6 months. The regenerated neo-corneas were transparent at 6 months post-operation. In vivo confocal microscopy confirmed normal morphology for the epithelium, stroma, sub-basal nerves and unoperated endothelium. Histology showed that all the regenerated corneas were morphologically similar to the normal. Immunohistochemistry indicated the presence of a differentiated corneal epithelium and functional tear film. In conclusion, the e-beamed corneal implants performed as well as non-irradiated control implants, resulting in fully regenerated neo-corneas with new nerves and without blood vessels or inflammation that may impede vision or corneal function. Therefore, a complete validation study to establish EB irradiation as an effective means for corneal implant sterilization prior to clinical application is necessary as a next step.https://www.frontiersin.org/articles/10.3389/fbioe.2022.883977/fullcollagenimplantE-beamirradiationrabbitscornea |
| spellingShingle | Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Fiona C. Simpson Mohammed Mirazul Islam Mohammed Mirazul Islam Oleksiy Buznyk Oleksiy Buznyk Elle Edin Elle Edin Elle Edin Elle Edin Elle Edin Marc Groleau Marc Groleau Monika Kozak-Ljunggren Federica M. Magrelli Dina B. AbuSamra Pablo Argüeso James Chodosh Aneta Liszka Per Fagerholm May Griffith May Griffith May Griffith May Griffith Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity Frontiers in Bioengineering and Biotechnology collagen implant E-beam irradiation rabbits cornea |
| title | Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity |
| title_full | Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity |
| title_fullStr | Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity |
| title_full_unstemmed | Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity |
| title_short | Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity |
| title_sort | electron beam irradiated recombinant human collagen phosphorylcholine corneal implants retain pro regeneration capacity |
| topic | collagen implant E-beam irradiation rabbits cornea |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.883977/full |
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