Experimental study of efficacy of intralamellar keratoplasty with corneal stromal substitute developed from decellularized porcine cornea
DOI:
https://doi.org/10.31288/oftalmolzh201734855Keywords:
intralamellar keratoplasty, porcine cornea, corneal stromal substitute, experimentAbstract
Backrgound: Enhancing methods for decellularization of animal corneas for the development of bioengineered corneal models with the following keratoplasty makes it possible to overcome the shortage of human donor material.
Purpose: To investigate the clinical efficacy of intralamellar keratoplasty (IKP) with corneal stromal substitutes (CSSs) developed using different decellularization methods in the animal study.
Materials and Methods: Corneas were excised from enucleated pig eyes, and, to develop acellular CSSs, five methods of decelluarization of porcine corneas were used that were different in the time and conditions of incubation with detergents (0.5% sodium dodecyl sulphate or TRITON X-100) and proteolytic enzymes (like 0.1% papain). In addition, an ultrasonic apparatus of type СD 3800 А was used to further improve the removal of cells and acellular corneal debris. A total of 40 Chinchilla rabbits were used in the animal study. They were divided into 5 groups with different CSS versions. In each rabbit, the monocular IKP was carried out with one of the five CSS versions. Postoperatively, antimicrobial and anti-inflammatory therapy was instituted. Animals were examined every other day for 30 days postoperative. This included corneal fluorescein staining. In addition, the following was assessed ophthalmoscopically: conjunctival discharge, conjunctival hyperemia, state of the CSS, state of the host cornea, and, at late follow-up examinations, graft acceptance and whether graft-versus-host disease was present.
Results: In rabbit eyes that underwent IKP with a version 4 CSS, the clinical scores for conjunctival discharge, corneal graft edema, inflammatory infiltration, corneal graft opacity and corneal fluorescein staining (0.13 points) were statistically significantly lower than in eyes that underwent IKP with other CSS versions. Location of inflammation in the cornea was found to be paracentral in eyes that improved clinically after IKP with version 1, 2, 3 or 5 CSS, and tended to be central (0.38 points) in those that underwent IKP with a version 4 CSS.
Conclusion: The least apparent reaction was noted in the rabbit eyes that underwent IKP with a version 4 CSS, which makes this CSS promising for further investigations in pre-clinical and clinical studies.
References
Dushin NV, Frolov MA, Gonchar PA. [Keratoplasty for optical, refractive, curative and esthetic indications in the treatment of ocular disorders: a tutorial]. Moscow: RUDN; 2008. Russian
Kaminski SL, Biowski R, Lukas JR, et al. Corneal sensitivity 10 years after epikeratoplasty. J Refract Surg. 2002 Nov-Dec;18(6):731-6 https://doi.org/10.3928/1081-597X-20021101-11
Whitcher JP, Srinivasan M, Upadhyay MP Corneal blindness: a global perspective. Bull WHO. 2001; 79: 214-21
Nasinnik IO. [Experimental substantiation for the use of corneal allograft]. [Abstract of Cand Sc (Med) Thesis]. Odessa: Filatov Institute of Eye Disease and Tissue Therapy; 2013. 19 p. Russian
Pasyechnikova NV, Vit VV, Leus NF, et al. [Clinical and histological results of experimental use of implants based on cross-linked collagen as donor cornea analogue]. Oftalmol Zh. 2011;4:58-60. Russian.
Pasyechnikova NV, Kogan BM, Gladush TI, et al. [The study of efficacy of in layer transplantation of heterogenous transplant of the cornea equivalent in the experiment]. Oftalmol Zh. 2014;2:64-70. Russian.
Choi JS, Williams JK, Greven M et al. Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma. Biomaterials. 2010 Sep;31(26):6738-45 https://doi.org/10.1016/j.biomaterials.2010.05.020
Cox A, Zhong R. Current advances in xenotransplantation. Hepatobiliary Pancreat Dis Int. 2005 Nov;4(4):490-4
A simple, cross-linked collagen tissue substitute for corneal implantation. Liu Y, Gan L, Carlsson DJ, et al. Invest Ophthalmol Vis Sci. 2006 May;47(5):1869-75 https://doi.org/10.1167/iovs.05-1339
Ponce M?rquez S, Mart?nez VS, McIntosh Ambrose W, et al. Decellularization of bovine corneas for tissue engineering applications. Acta Biomater. 2009 Jul;5(6):1839-47 https://doi.org/10.1016/j.actbio.2009.02.011
Pasyechnikova N., Vit V, Leus N, et al. Collagen-Based Bioengineered Substitutes of Donor Corneal Allograft Implantation: Assessment and Hypotheses. Med Hypothesis Discov Innov Ophthalmol. 2012 Spring; 1(1): 10-13
Rafat M, Li F, Fagerholm P, et al. PEG-stabilized carbodiimide crosslinked collagen-chitosanhydrogels for corneal tissue engineering. Biomaterials. 2008;29(29):3960-72 https://doi.org/10.1016/j.biomaterials.2008.06.017
Thompson RW Jr, Price MO, Bowers PJ, Price FW Jr. Long-term graft survival after penetrating keratoplasty. Ophthalmology. 2003 Jul;110(7):1396-402 https://doi.org/10.1016/S0161-6420(03)00463-9
Bartholomew LR, Pang DX, Sam DA, Cavender C. Ultrasound biomicroscopy of globes from young adult pigs. Am J Vet Res. 1997 Sep;58(9):942-8 https://doi.org/10.2460/ajvr.1997.58.09.942
Duan X, Sheardown H. Dendrimer crosslinked collagen as a corneal tissue engineering scaffold: mechanical properties and corneal epithelial cell interaction. Biomaterials. 2006 Sep;27(26):4608-17 https://doi.org/10.1016/j.biomaterials.2006.04.022
Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials. 2006 Jul;27(19):3675-83 https://doi.org/10.1016/j.biomaterials.2006.02.014
Wu Z, Zhou Y, Li N, et al. The use of phospholipase A2 to prepare acellular porcine corneal stroma as a tissue engineering scaffold. Biomaterials. 2009;30(21):3513-22 https://doi.org/10.1016/j.biomaterials.2009.03.003
Pasyechnikova NV, Vit VV, Leus NF, et al. [The reaction of the rabbit cornea after intralamellar transplantation of the acellular stroma of the human cornea]. Oftalmol Zh. 2011;1:57-60. Russian.
Yoeruek E, Bayyoud T, Maurus C, et al. Decellularization of porcine corneas and repopulation with human corneal cells for tissue-engineered xenografts. Acta Ophthalmol. 2012;90: e125-31 https://doi.org/10.1111/j.1755-3768.2011.02261.x
Wilson SL, Sidney LE, Dunphy SE, et al. Corneal Decellularization: A Method of Recycling Unsuitable Donor Tissue for Clinical Translation? Curr Eye Res. 2016 Jun;41(6):769-82 https://doi.org/10.3109/02713683.2015.1062114
Lu Y, Yao QK, Feng B, et al. Characterization of a Hydrogel Derived from Decellularized Corneal Extracellular Matrix. J Biomaterials and Tissue Engineering. 5: 951-60 https://doi.org/10.1166/jbt.2015.1410
Shafiq MA, Gemeinhart RA, Yue BY, Djalilian AR. Decellularized human cornea for reconstructing the corneal epithelium and anterior stroma. Tissue Eng Part C Methods. 2012 May;18(5):340-8 https://doi.org/10.1089/ten.tec.2011.0072
Hashimoto Y, Sasaki S, Hattori S, et al. Ultrastructural analysis of the decellularized cornea after interlamellar keratoplasty and microkeratome-assisted anterior lamellar keratoplasty in a rabbit model. Sci Rep. 2016; 6: 27734 https://doi.org/10.1038/srep27734
Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011 Apr;32(12):3233-43 https://doi.org/10.1016/j.biomaterials.2011.01.057
Hashimoto Y, Funamoto S, Sasaki S, et al. Preparation and characterization of decellularized cornea using high-hydrostatic pressurization for corneal tissue engineering. Biomaterials. 2010;31(14):3941-8 https://doi.org/10.1016/j.biomaterials.2010.01.122
Lee W, Miyagawa Y, Long C, et al. A comparison of three methods of decellularization of pig corneas to reduce immunogenicity. Int J Ophthalmol. 2014; 7(4): 587-93.
Yoeruek E, Bayyoud T, Maurus C, et al. Reconstruction of corneal stroma with decellularized porcine xenografts in a rabbit model. Acta Ophthalmol. 2012;90: e206-10 https://doi.org/10.1111/j.1755-3768.2011.02300.x
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Н. В. Пасечникова, Б. М. Коган, C. Г. Коломийчук

This work is licensed under a Creative Commons Attribution 4.0 International License.
This work is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) that allows users to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author as long as they cite the source.
COPYRIGHT NOTICE
Authors who publish in this journal agree to the following terms:
- Authors hold copyright immediately after publication of their works and retain publishing rights without any restrictions.
- The copyright commencement date complies the publication date of the issue, where the article is included in.
DEPOSIT POLICY
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) during the editorial process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work with an acknowledgement of its initial publication in this journal.
- Post-print (post-refereeing manuscript version) and publisher's PDF-version self-archiving is allowed.
- Archiving the pre-print (pre-refereeing manuscript version) not allowed.








