Experimental study of efficacy of intralamellar keratoplasty with corneal stromal substitute developed from decellularized porcine cornea

Authors

  • N.V. Pasyechnikova SI "The Filatov Institute of Eye Diseases and Tissue Therapy of the NAMS of Ukraine"; Odesa (Ukraine)
  • B.M. Cogan SI "The Filatov Institute of Eye Diseases and Tissue Therapy of the NAMS of Ukraine"; Odesa (Ukraine)
  • S.G. Kolomiichuk SI "The Filatov Institute of Eye Diseases and Tissue Therapy of the NAMS of Ukraine"; Odesa (Ukraine)

DOI:

https://doi.org/10.31288/oftalmolzh201734855

Keywords:

intralamellar keratoplasty, porcine cornea, corneal stromal substitute, experiment

Abstract

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

Published

2026-02-06

How to Cite

[1]
Pasyechnikova, N. et al. 2026. Experimental study of efficacy of intralamellar keratoplasty with corneal stromal substitute developed from decellularized porcine cornea. Ukrainian Journal of Ophthalmology . 3 (Feb. 2026), 48–55. DOI:https://doi.org/10.31288/oftalmolzh201734855.

Issue

Section

Experimental Studies

Most read articles by the same author(s)

<< < 1 2 3 > >> 

Similar Articles

1 2 3 4 5 > >> 

You may also start an advanced similarity search for this article.