https://doi.org/10.31288/oftalmolzh201613642

Corneal inflammation and proliferative activity of cells of anterior epithelium in modeling bacterial keratitis and using amniotic membrane fixed with different techniques

E.V. Sereda

V.V. Vit, Dr. Sc. (Med), Prof

G.I. Drozhzhina, Dr. Sc. (Med), Prof

T.B. Gaidamaka, Dr. Sc. (Med)

Filatov Institute of Eye Diseases and Tissue Therapy

Odessa, Ukraine

E-mail: evsereda08@gmail.com                           

Introduction. Molecular mechanisms of the effect of amniotic membrane (AM) on the inflammation and recover of the cornea have been investigated using various biological markers.

Purpose. To determine the dependence between corneal inflammation and proliferative activity of anterior epithelium cells studying the expressions of Ki-67, CD-68, ММР-9 in a model simulating bacterial keratitis and when using amniotic membrane fixed with different techniques.

Material and Methods. 60 chinchilla rabbits (60 eyes) were involved into the experimental study. The animals were induced moderate bacterial keratitis. In two weeks, the rabbits were performed AM transplantation; two techniques were used: an inlay technique with AM secured to the cornea with eight interrupted 10/00 nylon sutures, and an onlay technique with AM sutured to episclera using eight interrupted 8/00 silk sutures.

Results. The study revealed the presence of negative correlation between a proliferative activity of anterior corneal epithelial cells and a degree of inflammatory infiltration of the corneal stroma as evidenced by studying such markers of corneal repair as Ki-67, CD 68 and MMP-9. If inflammation occurs, the proliferative activity of anterior epitehelium cells is reduced that significantly decreases epithelization rates and following lamellar cell differentiation.

Conclusions. Inflammation and epithelization rate reduction occurs more often when AM is sutured to the corneal surface, i.g. in group of experimental animals, in which the inlay technique of AMT was used.

Key words:bacterial keratitis, amniotic membrane, experiment, biological markers, repair 

     

References

1.Hao Y, Ma DH, Hwang DG, Kim WS, Zhang F.  Identification of antiangiogenic and antiinflammatory proteins in human amniotic membrane. Cornea. 2000;19:348-52.

2.Kjaergaard N, Hein M, Hyttel L et al. Antibacterial properties of human amnion and chorion in vitro. Eur J Obstet Gynecol Reprod Biol. 2001;94:224–9.

3.Talmi WP, Sigler L, Inge E, Finkelstein Y, Zohar Y. Antibacterial properties of human amniotic membranes. Placenta. 1991;12:285-8.

4.Tseng SC, Li DQ, Ma X. Suppression of transforming growth factor-beta isoforms, TGF-beta receptor type II, and myofibroblast differentiation in cultured human corneal and limbal fibroblasts by amniotic membrane matrix. J. Cell Physiol. 1999;179:325-35.

5.Heiligenhaus A, Li  H, Yang Y, Wasmuth S, Bauer  D, Steuhl KP. Amniotic membrane transplantation improves experimental herpetic keratitis. Modulation of matrix metalloproteinase-9. Ophthalmologe. 2004;101(1):59-65. In German.

6.Bauer D, Wasmuth S, Hennig M, Baehler H, Steuhl KP, Heiligenhaus  A. Amniotic membrane transplantation induces apoptosis in T lymphocytes in murine corneas with experimental herpetic stromal keratitis. Invest. Ophthalmol. Vis. Sci. 2009;50(7):3188-98.

7.Barequet Irina S, Habot-Wilner Z, Keller N,  Smollan G, Ziv H, Belkin M,  Rosner M. Effect of Amniotic Membrane Transplantation on the Healing of Bacterial Keratitis. IOVS. Jan 2008;49:163-7.

8.Gicquel JJ, Bejjani RA, Ellies P, Mercie M, Dighiero P. Amniotic membrane transplantation in severe bacterial keratitis. Cornea. 2007;26:27-33.

9.Kim JS, Hahn TW, Park WC. Amniotic membrane transplantation in infectious corneal ulcer. Cornea. 2001;20:720–6.

10.Bauer D, Hennig  M, Wasmuth  S, Baehler  H, Busch  M, Steuhl  KP, Thanos  S, Heiligenhaus  A. Amniotic membrane induces peroxisome proliferator-activated receptor-? positive alternatively activated macrophages. Invest Ophthalmol Vis Sci.  2012;53(2):799-810. 

11.Bromley M, Rew D, Becciolini A et al.  A comparison of proliferation markers (BrdUrd, Ki-67. PCNA) determined at each cell position in the crypts of normal human colonic mucosa. Eur. J. Histochem. 1996;40:89-100.

12.Smorodinova N, Kaltner  H, Jirsov?  K, Hrdli?kov?-Cela E, Andr? S, Ku?era T, Smetana K Jr., Gabius  HJ. Regulatory Impact of Amniotic Membrane Transplantation on Presence of Adhesion/Growth-Regulatory Galectins-1 and -7 in Corneal Explants from Acanthamoeba Keratitis Patients: Clinical Note. Curr Eye Res. 2015;4:1-7.

13.Bruno S, Darynkiewich Z. Cell cycle dependent expression and stability of the nuclear protein by Ki-67 antibody in HL-60 cells. Cell. Prolif. 1992;25:31-40.

14.Holt PR, Moss SF, Kapetanakis AM. Is Ki-67 a better proliferative marker in the colon then proliferating cell nuclear antigen? Cancer. Epidimiol. Biomarkers. Prev. 1997;6:131-5.

15.Dong Z,  Zhou X,  Wu J,  Zhang Z et al. Small incision lenticule extraction (SMILE) and femtosecond laser LASIK: comparison of corneal wound healing and inflammation. Br J Ophthalmol. 2014;98(2):263-9.

16.Fabiani С,  Barabino S,  Rashid S, Reza Dana M. Corneal Epithelial Proliferation and Thickness in a Mouse Model of Dry Eye.  Exp Eye Res. 2009;89(2):1166-1171.

17.Tandon R, Gupta N, Kalaivani M, Sharma N, Titiyal JS, Vajpayee RB. Amniotic membrane transplantation as an adjunct to medical therapy in acute ocular burns. Br J Ophthalmol. 2011;95:199-204.

18.Tseng SC, Di Pascuale MA, Liu DT, Gao YY, Baradaran-Rafii A. Intraoperative mitomycin C and amniotic membrane transplantation for fornix reconstruction in severe cicatricial ocular surface diseases. Ophthalmology. 2005;112:896–903.

19.Kenney MC, Chwa M, Lin B, Huang GH, Ljubimov AV, Brown  DJ. Identification of cell types in human diseased corneas. Cornea. 2001;20(3):309-16.

20.Liu T, Zhai H, Xu Y, Dong Y, Sun Y, Zang X, Zhao J. Amniotic membrane traps and induces apoptosis of inflammatory cells in ocular surface chemical burn. Mol Vis.  2012;18 (2):137-46.

21.Ollivier FJ, Gilger BC, Barrie KP et al. Proteinases of the cornea and preocular tear film. Vet Ophthalmol. 2007;199–206.

22.Xiaoyong Y, Mitchell BM, Wilhelmus KR. Expression of Matrix Metalloproteinases during Experimental Candida albicans Keratitis. Invest Ophthalmol Vis Sci. 2009;50(2):737-42. 

23.Sereda EV, Drozhzhyna GI, Gaidamaka TB. Экспериментальная модель бактериального кератита средней степени тяжести. Oftalmologiia. Vostochnaia Evropa. 2015;2(25); 41-8. In Russian.