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  • 1.
    Bentley, E
    et al.
    Univ Wisconsin, Dept Surg Sci, Sch Vet Med, Madison, WI 53706 USA.
    Murphy, CJ
    Univ Wisconsin, Dept Surg Sci, Sch Vet Med, Madison, WI 53706 USA.
    Li, FF
    Univ Ottawa, Inst Eye, Ottawa, ON Canada.
    Carlsson, DJ
    Univ Ottawa, Inst Eye, Ottawa, ON Canada.
    Griffith, May
    Univ Ottawa, Inst Eye, Ottawa, ON Canada.
    Biosynthetic Corneal Substitute Implantation in Dogs2010Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 29, nr 8, s. 910-916Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: To assess integration of a biosynthetic corneal implant in dogs.

    Methods: Three normal adult laboratory Beagles underwent ophthalmic examinations, including slit-lamp biomicroscopy, indirect ophthalmoscopy, applanation tonometry, and Cochet-Bonnet aesthesiometry. Biosynthetic corneas fabricated from glutaraldehyde crosslinked collagen and copolymers of collagen and poly (N-isopropylacrylamide-co-acrylic acid-co-acryloxysuccinimide, denoted as TERP) were implanted into dogs by a modified epikeratoplasty technique. Ophthalmic examinations and aesthesiometry were performed daily for 5 days and then weekly thereafter for 16 weeks. Corneal samples underwent histopathological and transmission electron microscopy examination at 16 weeks.

    Results: Implants were epithelialized by 7 days. Intraocular pressure was within normal range throughout the study. Aesthesiometry values dropped from an average of 3.67 cm preoperatively to less than 1 mm for all dogs for the first postoperative weeks. By week 16, the average Cochet-Bonnet value was 1.67 cm, demonstrating partial recovery of functional innervation of the implant. No inflammation or rejection of the implant occurred, and minimal haze formation was noted. Light microscopy revealed thickened but normal epithelium over the implant with fibroblast migration into the scaffold. On transmission electron microscopy, the basement membrane was irregular but present and adhesion complexes were noted.

    Conclusion: Biosynthetic corneal implantation is well tolerated in dogs, and the collagen-polymer hybrid construct holds promise for clinical application in animals and humans.

  • 2.
    Gan, Lisha
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Östergötlands Läns Landsting, Rekonstruktionscentrum, Ögonkliniken US/LiM.
    Fagerholm, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Östergötlands Läns Landsting, Rekonstruktionscentrum, Ögonkliniken US/LiM.
    Leukocytes in the early events of corneal neovascularization2001Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 20, s. 96-99Artikel i tidskrift (Refereegranskat)
  • 3.
    Griffith, May
    et al.
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Hakim, Malik
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada and National Research Council of Canada - ICPET, Ottawa, Ontario, Canada.
    Shimmura, Shigeto
    Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Watsky, Mitchell A.
    Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee, USA.
    Li, Fengfu
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada and National Research Council of Canada - ICPET, Ottawa, Ontario, Canada.
    Carlsson, David
    National Research Council of Canada - ICPET, Ottawa, Ontario, Canada.
    Doillon, Charles J.
    Oncology and Molecular Endocrinology Research Center, Laval University, Quebec, Canada.
    Nakamura, Masatsugu
    Santen Pharmaceutical Co. Ltd., Nara R&D Center, Ikoma-shi, Nara, Japan.
    Suuronen, Erik
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Shinozaki, Naoshi
    Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Nakata, Katsuhiko
    Santen Pharmaceutical Co. Ltd., Nara R&D Center, Ikoma-shi, Nara, Japan.
    Sheardown, Heather
    Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
    Artificial human corneas - Scaffolds for transplantation and host regeneration2002Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 21, nr 7, s. S54-S61Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose. To review the development of artificial corneas (prostheses and tissue equivalents) for transplantation, and to provide recent updates on our tissue-engineered replacement corneas. Methods. Modified natural polymers and synthetic polymers were screened for their potential to replace damaged portions of the human cornea or the entire corneal thickness. These polymers, combined with cells derived from each of the three main corneal layers or stem cells, were used to develop artificial corneas. Functional testing was performed in vitro. Trials of biocompatibility and immune and inflammatory reactions were performed by implanting the most promising polymers into rabbit corneas. Results. Collagen-based biopolymers, combined with synthetic crosslinkers or copolymers, formed effective scaffolds for developing prototype artificial corneas that could be used as tissue replacements in the future. We have previously developed an artificial cornea that mimicked key morphologic and functional properties of the human cornea. The addition of synthetic polymers increased its toughness as it retained transparency and low light scattering, making the matrix scaffold more suitable for transplantation. These new composites were implanted into rabbits without causing any acute inflammation or immune response. We have also fabricated full-thickness composites that can be fully sutured. However, the long-term effects of these artificial corneas need to be evaluated. Conclusions. Novel tissue-engineered corneas that comprise composites of natural and synthetic biopolymers together with corneal cell lines or stem cells will, in the future, replace portions of the cornea that are damaged. Our results provide a basis for the development of both implantable temporary and permanent corneal replacements.

  • 4.
    McLaughlin, Christopher
    et al.
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Fagerholm, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för klinisk och experimentell medicin, Oftalmiatrik. Östergötlands Läns Landsting, Rekonstruktionscentrum, Ögonkliniken US.
    Muzakare, Lea
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Lagali, Neil
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Forrester, John
    Department of Ophthalmology University of Aberdeen, Aberdeen, Scotland, United Kingdom.
    Kuffova, Lucia
    Department of Ophthalmology University of Aberdeen, Aberdeen, Scotland, United Kingdom.
    Rafat, Mehrdad
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Liu, Yuwen
    National Research Council Canada, Ottawa, Ontario, Canada.
    Shinozaki, Naoshi
    Tokyo Dental College-Ichikawa General Hospital Cornea Centre, Ichikawa, Chiba, Japan.
    Vascotto, Sandy
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Munger, Rejean
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Griffith, May
    University of Ottawa Eye Institute, Ottawa, Ontario, Canada.
    Regeneration of Corneal Cells and Nerves in an Implanted Collagen Corneal Substitute2008Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 27, nr 5, s. 580-589Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    PURPOSE: Our objective was to evaluate promotion of tissue regeneration by extracellular matrix (ECM) mimics, by using corneal implantation as a model system.

    METHODS: Carbodiimide cross-linked porcine type I collagen was molded into appropriate corneal dimensions to serve as substitutes for natural corneal ECM. These were implanted into corneas of mini-pigs after removal of the host tissue, and tracked over 12 months, by clinical examination, slit-lamp biomicroscopy, in vivo confocal microscopy, topography, and esthesiometry. Histopathology and tensile strength testing were performed at the end of 12 months. Other samples were biotin labeled and implanted into mice to evaluate matrix remodeling.

    RESULTS: The implants promoted regeneration of corneal cells, nerves, and the tear film while retaining optical clarity. Mechanical testing data were consistent with stable, seamless host-graft integration in regenerated corneas, which were as robust as the untreated fellow corneas. Biotin conjugation is an effective method for tracking the implant within the host tissue.

    CONCLUSIONS: We show that a simple ECM mimetic can promote regeneration of corneal cells and nerves. Gradual turnover of matrix material as part of the natural remodeling process allowed for stable integration with host tissue and restoration of mechanical properties of the organ. The simplicity in fabrication and shown functionality shows potential for ECM   

  • 5.
    Podskochy, A.
    et al.
    St. Erik's Eye Hospital, Karolinska Institutet, Stockholm, Sweden.
    Fagerholm, P.
    St. Erik's Eye Hospital, Karolinska Institutet, Stockholm, Sweden.
    The expression of fas ligand protein in ultraviolet-exposed rabbit corneas2002Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 21, nr 1, s. 91-94Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose. Keratocytes undergo apoptosis during various pathologic conditions and after exposure to ultraviolet radiation (UVR). It was reported that the Fas/Fas ligand system is involved in modulating keratocyte apoptosis. The expression of Fas ligand (FasL) protein was studied in rabbit corneas after photokeratitis induced by different UV wavelengths. Methods. Six New Zealand albino rabbit corneas were exposed to 280- and 310-nm UVR in 10-nm full wavebands at doses producing biomicroscopically significant keratitis (0.12 J/cm2 for 280 nm and 0.47 J/cm2 for 310 nm). Animals were killed 24 hours after exposure. Immunohistochemistry was performed to localize FasL protein in paraffin sections of rabbit corneas. Primary antibody was polyclonal goat anti-FasL IgG. Results. FasL protein was uniformly detected in epithelial and endothelial layers of all UVR-exposed and control, nonexposed corneas. The positive staining of keratocytes was confined to the anterior stroma of corneas that were exposed to 280-nm UVR. Corneas exposed to 310-nm UVR showed positively stained keratocytes throughout the entire thickness of the stroma. Conclusions. These data strongly suggest that the Fas/FasL system may play an important role in apoptosis of corneal cells after UVR. The FasL expression in the corneal stroma was more extensive after exposures at 310-nm UVR than at 280-nm UVR.

  • 6.
    Samarawickrama, Chameen
    et al.
    Moorfields Eye Hosp NHS Fdn Trust, England; UCL Inst Ophthalmol, England; Univ Sydney, Australia.
    Samanta, Ayan
    Uppsala Univ, Sweden; LV Prasad Eye Inst, India.
    Liszka, Aneta
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för cellbiologi. Linköpings universitet, Medicinska fakulteten.
    Fagerholm, Per
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. Region Östergötland, Sinnescentrum, Ögonkliniken US/LiM.
    Buznyk, Oleksiy
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för neuro- och inflammationsvetenskap. Linköpings universitet, Medicinska fakulteten. NAMS Ukraine, Ukraine.
    Griffith, May
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för cellbiologi. Linköpings universitet, Medicinska fakulteten. LV Prasad Eye Inst, India; Univ Montreal, Canada; Univ Montreal, Canada.
    Allan, Bruce
    Moorfields Eye Hosp NHS Fdn Trust, England; UCL Inst Ophthalmol, England.
    Collagen-Based Fillers as Alternatives to Cyanoacrylate Glue for the Sealing of Large Corneal Perforations2018Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 37, nr 5, s. 609-616Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: To describe the use of collagen-based alternatives to cyanoacrylate glue for the sealing of acute corneal perforations. Methods: A collagen analog comprising a collagen-like peptide conjugated to polyethylene glycol (CLP-PEG) and its chemical crosslinker were tested for biocompatibility. These CLP-PEG hydrogels, which are designed to act as a framework for corneal tissue regeneration, were then tested as potential fillers in ex vivo human corneas with surgically created full-thickness perforations. Bursting pressures were measured in each of 3 methods (n = 10 for each condition) of applying a seal: 1) cyanoacrylate glue with a polyethylene patch applied ab externo (gold standard); 2) a 100-mu m thick collagen hydrogel patch applied ab interno, and 3) the same collagen hydrogel patch applied ab interno supplemented with CLP-PEG hydrogel molded in situ to fill the remaining corneal stromal defect. Results: Cyanoacrylate gluing achieved a mean bursting pressure of 325.9 mm Hg, significantly higher than the ab interno patch alone (46.3 mm Hg) and the ab interno patch with the CLP-PEG filler (86.6 mm Hg). All experimental perforations were sealed effectively using 100 mu m hydrogel sheets as an ab interno patch, whereas conventional ab externo patching with cyanoacrylate glue failed to provide a seal in 30% (3/10) cases. Conclusions: An ab interno patch system using CLP-PEG hydrogels designed to promote corneal tissue regeneration may be a viable alternative to conventional cyanoacrylate glue patching for the treatment of corneal perforation. Further experimentation and material refinement is required in advance of clinical trials.

  • 7.
    Shimmura, Shigeto
    et al.
    Department of Ophthalmology, Tokyo Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Doillon, Charles J.
    Laval University Medical Center (CHUL), Laval University, Quebec, Canada.
    Griffith, May
    University of Ottawa Eye Institute, University of Ottawa, Ottawa, Ontario, Canada.
    Nakamura, Masatsugu
    Santen Pharmaceutical Co. Ltd., Nara R&D Center, Ikoma-shi, Nara, Japan.
    Gagnon, Edith
    Laval University Medical Center (CHUL), Laval University, Quebec, Canada.
    Usui, Akemi
    Department of Ophthalmology, Tokyo Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Shinozaki, Naoshi
    Department of Ophthalmology, Tokyo Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Tsubota, Kazuo
    Department of Ophthalmology, Tokyo Dental College, Ichikawa General Hospital Cornea Center, Ichikawa, Chiba, Japan.
    Collagen-poly(N-isopropylacrylamide)-based membranes for corneal stroma scaffolds2003Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 22, nr 7, s. S81-S88Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: To investigate the feasibility of using the biocompatibility of collagen-based blended biomaterials as cell-delivery systems in ocular surface reconstruction in vivo. Methods: Collagen-based composites that were blended with synthetic acrylamide-based polymers [poly(N-isopropylacrylamide), pNIPAAm] were transplanted into corneal pockets of white rabbits, with a 3-mm epithelial window. Epithelial cells were allowed to migrate onto the polymer. Transplanted eyes were examined daily for up to 30 days, after which animals; were killed for histologic examination. lm- munohistochemistry was performed for vimentin, a-smooth muscle actin (alpha-SMA), CD4, and CD8. Gold-chloride staining was performed to observe neuronal regrowth. Human amniotic membranes (AMs) and sham-operated corneas served as controls. All animals received topical antibiotics (levofloxacin) without the use of steroids or other immunosuppressive agents. Results: The pNIPAAm polymer allowed smooth epitheliatization of the cornea, which was similar to the epithelialization observed in sham controls and AM-transplanted eyes. Histology revealed that epithelium overlying the polymer was bundled into several layers, without the orientation observed with AM and sham controls. The polymer gradually thinned and was gradually replaced by host tissue. Vimentin- and alpha-SMA-positive cells were found in stromal pockets up to 1 month following polymer transplantation. These cells were responsible for slight subepithelial haze near the wound edge. CD4- and CD8-positive lymphocytes were also observed in the vicinity of the polymer. Gold-chloride staining showed nerve regrowth in the wound edge after 1 month and subepithelial branches after 3 months. Conclusion: Collagen-pNIPAAm blended polymers may he effective as biomaterials to be used in the early stages of lamellar stromal replacement.

  • 8.
    Wand, Kerstin
    et al.
    Technical University of Munich, Germany.
    Neuhann, Raphael
    Technical University of Munich, Germany.
    Ullmann, Andrea
    Technical University of Munich, Germany.
    Plank, Katharina
    Technical University of Munich, Germany.
    Baumann, Michael
    MLase AG, Germany.
    Ritter, Roland
    Technolas PV Bausch and Lomb, Germany.
    Griffith, May
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Avdelningen för cellbiologi. Linköpings universitet, Hälsouniversitetet.
    Lohmann, Chris P.
    Technical University of Munich, Germany.
    Kobuch, Karin
    Technical University of Munich, Germany.
    Riboflavin-UV-A Crosslinking for Fixation of Biosynthetic Corneal Collagen Implants2015Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 34, nr 5, s. 544-549Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose: To evaluate riboflavin-UV-A crosslinking as an alternative suture-free fixation method for biosynthetic corneal collagen implants. Methods: A range of cell-free corneal implants consisting of recombinant human collagen type III were examined. In vitro, the implants were crosslinked with different riboflavin solutions and irradiations. Ex vivo, the biosynthetic corneal implants were placed on the anterior cornea of porcine and rabbit eyes after performing deep anterior lamellar keratoplasty with a trephine, femtosecond laser, or excimer laser. UV-A crosslinking was performed with isotonic or hypotonic riboflavin at either standard or rapid procedure. The corneas were excised, fixed in PFA 4%, and embedded in paraffin. Crosslinking effects on the implants and the adhesion between implant and corneal bed were evaluated by slit-lamp biomicroscopy, optical coherence tomography (OCT) images, and histologically. Results: After the crosslinking procedure, the implants showed different degrees of thinning. The accuracy of cutting the corneal bed was highest with the excimer laser. Good adhesion of the implant in the corneal bed could be demonstrated in OCT images. This was more accurate in porcine eyes than in rabbit eyes. Histologically, crosslinks between implant and corneal stroma were demonstrated. There was no difference between standard and rapid crosslinking procedures. Conclusions: Riboflavin-UV-A crosslinking as a fixation method for biosynthetic corneal collagen implants was demonstrated to be promising. It can reduce suture-related complications such as haze formation and surface irregularity. Stability of the implants, especially shrinkage after riboflavin-UV-A crosslinking, needs to be further evaluated. Biostability, integration, and long-term outcome are further evaluated in in vivo animal experiments.

  • 9. Weber, Beat A
    et al.
    Gan, Lisha
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Östergötlands Läns Landsting, Rekonstruktionscentrum, Ögonkliniken US/LiM.
    Fagerholm, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Oftalmologi. Östergötlands Läns Landsting, Rekonstruktionscentrum, Ögonkliniken US/LiM.
    Short-term impact of corticosteroids on hyaluronan and epithelial hyperplasia in the rabbit cornea after photorefractive keratectomy2001Ingår i: Cornea, ISSN 0277-3740, E-ISSN 1536-4798, Vol. 20, nr 3, s. 321-324Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Purpose. To investigate the impact of corticosteroids on subepithelial hyaluronan deposition and corneal epithelium thickness in the first 10 days after photorefractive keratectomy (PRK) and to analyze a possible contralateral effect of corticosteroids. Methods. Thirty-two New Zealand white rabbits were assigned into two groups and had a transepithelial 5.0-mm diameter, 8.00-diopter myopic PRK performed on one eye. The corticosteroid treatment group (16 animals) received 0.1 mL of methylprednisolone 4% subconjunctivally on the operation table, followed by 0.1% dexamethasone eye drops six times a day during the postoperative period. The sodium chloride (NaCl) treatment group received topical isotonic NaCl eye drops six times a day. In each treatment group, eight animals were killed after 3 and 9 days, respectively. The harvested specimens were stained for hyaluronan and the epithelial thickness was measured. Results. In contrast to the epithelial thickness, the subepithelial hyaluronan did not show a significant increase during the observation period. The corticosteroid treated group showed at both time-points significantly less subepithelial hyaluronan formation as well as a significantly thinner epithelium, when compared with the NaCl-treated group. At 9 days, the corticosteroid-treated group showed a mild epithelial hyperplasia in only one of eight eyes, whereas this was a common finding in the NaCl-treated group. We detected no hyaluronan deposits in any contralateral-untreated eye, and the epithelial thickness did not differ significantly between any of the four contralateral-untreated eye groups. Conclusions. Corticosteroid medication during the first 10 days after operation reduces the amount of subepithelial hyaluronan production and inhibits the epithelial proliferation, and epithelial hyperplasia is prevented. Neither a contralateral hyaluronan deposition nor a contralateral corticosteroid effect could be detected.

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