Oftalmol Zh.2015;1:110-114

https://doi.org/10.31288/oftalmolzh20151110114

Collagen-Based Corneal Substitutes with Incorporated Anti-infective Peptide LL37 Sustalined Deivery System 

N. V. Pasyechnikova1, S. A. Yakymenko1, O. I. Buznyk1, M. M.  Islam2, M. Griffith

1 State Institution The Filatov Institute of Eye Diseases and Tissue Therapy of the NAMS of Ukraine; Odessa, (Ukraine) 

2 Linkoping University; Linkoping (Sweden)

Purpose. To develop corneal substitutes (CS) with a dual function:1) corneal stroma substitute, and 2) antiinfective peptide (AIP) delivery. 

Methods. AIP LL37 (cathelicidin) was encapsulated in silica nanoparticles (SINP) under the effect of magnetic field. SiNP with LL37 were then introduced in CS during their production using the method of creation of interpenetrating network of type I collagen and 2 methacryloyloxyethylphosphorylcholine. Dynamics of LL37 release from CS using ELISA method was evaluated. Refractive properties of CS with incorporated AIP release system were assessed using Abbe refractometer. Cytotoxicity of the CS to the cells of the human corneal epithelium was assessed by using WST-1 based colorimetric assay. Mechanical properties were checked by performing lamellar keratoplasty on ex vivo porcine eyes. 

Results. Gradual release of LL37 from within CS occurred up to 21day. Release from control CS (with incorporated free LL37) stopped on the 1st day. Refractive index (RI) of CS with LL37 in SiNP was equal to 1.3 (human cornea 1.37). There was no increased cytotoxicity of CS with incorporated release system compared to clear CS. Mechanical strength of CS with LL37 was non-significantly lower compared to clear CS. 

Conclusion. For the first time CS containing AIP sustained delivery system was developed. Gradual release of AIP LL37 from within CS was proved. No cell toxicity, satisfactory optical and mechanical properties of the constructs were found. 

Key words: artificial cornea, antiinfective peptide LL37, sustained delivery system

References 

1.Buznyk OI. The sustained delivery system of the anti-infection peptide LL37 system — a potentially new treatment method of ocular infections. Report 1. Testing of different nano- and microparticles as carriers of LL37. Oftalmol Zh. 2014;2:17–21. In Russian. 

Crossref  

2.Gaidamaka TB. Recurrent herpetic keratitis. Pathogenesis, diagnosis, treatment, and prevention: Author’s thesis for Doctor of Med. Science. 14.01.18. Ophthalmology. SI «The Filatov Institute of Eye Diseases and Tissue Therapy of NAMS of Ukraine». Odessa, 2011. 44 p. 

3.Islam M. M., Griffith M., Mondal D., Buznyk O. I. Anti-infection peptide LL37 sustained delivery system — a potential new treatment of ocular infections. Report 2. Antiviral activity of LL37 incapsulated in silica nanoparticles. Oftalmol Zh. 2014;3:53–7. 

Crossref 

4.Pasyechnikova N, Buznyk A, Yakymenko S et al. Sustained delivery system of the antiinfection peptide LL37 — a potential new method of treat¬ment of ocular infections. Report 3. Antimicrobial activity of LL37 encapsulated in silica nanoparticle. Oftalmol Zh. In printing. 

5.Bareiss B, Ghorbani M, Li F et al. Controlled release of acyclovir through bioengineered corneal implants with silica nanoparticle carriers. B. Bareiss. Open Tissue Eng. Regen. Med. J. 2010;3:10–7. 

Crossref  

6.Donadio S, Maffioli S, Monciardini P, Sosio M, Jabes D. Antibiotic discovery in the twenty-first century: Current trends and future perspectives. J. Antibiot. (Tokyo). 2010;63(8):423–30. 

Crossref   PubMed  

7.Fagerholm P, Lagali NS, Merrett K et al. A biosynthetic alternative to human donor tissue for inducing corneal regeneration: 24-month follow-up of a phase 1 clinical study. Sci. Transl. Med. 2010;2:46–61. 

Crossref   PubMed  

8.Gordon YJ, Huang LC, Romanowski EG et al. Human cathelicidin (LL-37), a multifunctional peptide, is expressed by ocular surface epithelia and has potent antibacterial and antiviral activity. 2005;30(5):385–94. 

Crossref   PubMed  

9.Griffith M, Polisetti N, Kuffova L et al. Regenerative approaches as alternatives to donor allografting for restoration of corneal function. Ocul. Surf. 2012;10(3): 170–83. 

Crossref  PubMed

10.Liu W, Deng C, McLaughlin CR et al. Collagen-phosphorylcholine interpenetrating network hydrogels as corneal substitutes. Biomaterials. 2009;30:1551–9. 

Crossref   PubMed 

11.Pasyechnikova N, Vit V, Leus M et al. Collagen-based bioengineered substitutes of donor corneal allograft implantation: assessment and hypotheses. Med. Hypothesis Discov. Innov. Ophthalmol. 2012;1(1);10–3. 

12.Sangwan VS, Gopinathan U, Garg P, Rao GN. Eye Banking in India: A Road Ahead. JIMSA.2010;23(3):197–200. 

13.Sharma N, Sachdev R, Jhanji V et al. Therapeutic keratoplasty for microbial keratitis. Curr. Opinion Ophthalmol. 2010;21:293–300. 

Crossref   PubMed 

14.Whitcher JP, Srinivasan M, Upadhyay MP. Corneal blindness: a global perspective. Bull. World Health Organ. 2001;79:214–21.