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Pharmaceutical Research

, Volume 34, Issue 5, pp 1083–1092 | Cite as

Ocular safety of Intravitreal Clindamycin Hydrochloride Released by PLGA Implants

  • Gabriella M. Fernandes-CunhaEmail author
  • Silvia Ligório Fialho
  • Gisele Rodrigues da Silva
  • Armando Silva-Cunha
  • Min Zhao
  • Francine Behar-Cohen
Research Paper

ABSTRACT

Background

Drug ocular toxicity is a field that requires attention. Clindamycin has been injected intravitreally to treat ocular toxoplasmosis, the most common cause of eye posterior segment infection worldwide. However, little is known about the toxicity of clindamycin to ocular tissues. We have previously showed non intraocular toxicity in rabbit eyes of poly(lactic-co-glycolic acid) (PLGA) implants containing clindamycin hydrochloride (CLH) using only clinical macroscotopic observation. In this study, we investigated the in vivo biocompatibility of CLH-PLGA implants at microscotopic, cellular and molecular levels.

Methods

Morphology of ARPE-19 and MIO-M1 human retinal cell lines was examined after 72 h exposure to CLH-PLGA implant. Drug delivery system was also implanted in the vitreous of rat eyes, retinal morphology was evaluated in vivo and ex vivo. Morphology of photoreceptors and inflammation was assessed using immunofluorescence and real-time PCR.

Results

After 72 h incubation with CLH-PLGA implant, ARPE-19 and MIO-M1 cells preserved the actin filament network and cell morphology. Rat retinas displayed normal lamination structure at 30 days after CLH-PLGA implantation. There was no apoptotic cell and no loss in neuron cells. Cones and rods maintained their normal structure. Microglia/macrophages remained inactive. CLH-PLGA implantation did not induce gene expression of cytokines (IL-1β, TNF-α, IL-6), VEGF, and iNOS at day 30.

Conclusion

These results demonstrated the safety of the implant and highlight this device as a therapeutic alternative for the treatment of ocular toxoplasmosis.

KEY WORDS

biocompatibility clindamycin intravitreal implant ocular toxoplasmosis PLGA toxicity 

ABBREVIATIONS

ARPE-19

Human retinal pigment epithelial cell line

CLH

Clindamycin hydrochloride

DAPI

2-(4-amidinophenyl)-1H -indole-6-carboxamidine

DMEM/F-12

Dulbecco’s modified eagle medium: nutrient mixture F-12

ERG

Electroretinogram

FBS

Fetal bovine serum

FITC

Fluorescein isothiocyanate

GCL

Ganglion cell layer

HAM

Human amniotic membrane

HEPES

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HPRT

Hypoxanthine phosphoribosyltransferase

IBA-1

Ionized calcium binding adaptor molecule 1

IL-1β

Interleukin 1 beta

IL-6

Interleukin 6

INL

Inner nuclear layer

iNOS

Inducible nitric oxide synthase

IP

Propidium iodide

MIO-M1

Human Müller cell line

OCT

Optical coherence tomography

OLN

Outer nuclear layer

PBS

Phosphate-buffered saline

PCR

Polymerase chain reaction

PLGA

Poly(lactic-co-glycolic acid)

PNA

Peanut Agglutinin

SD

Standard deviation

TNF-α

Tumor necrosis factor alpha

TUNEL

Terminal deoxynucleotidyl transferase dUTP nick end labeling

VEGF

Vascular endothelial growth factor

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

The authors would like to acknowledge the financial support received from the following institutions: (Brazil), FAPEMIG (Minas Gerais – Brazil), Pró-reitoria de Pesquisa da Universidade Federal de Minas Gerais (Minas Gerais – Brazil), CAPES (Bolsistas da CAPES-Brasília/Brazil).

REFERENCES

  1. 1.
    Saliba JB, Vieira L, Fernandes-Cunha GM, Silva GR, Fialho SL, Silva-Cunha A, et al. Anti-inflammatory effect of dexamethasone controlled released from anterior suprachoroidal polyurethane implants on endotoxin-induced uveitis in rats. Invest Ophthalmol Vis Sci. 2016;57(4):1671–9.CrossRefGoogle Scholar
  2. 2.
    Elsaesser A, Howard CV. Toxicology of nanoparticles. Adv Drug Deliv Rev. 2012;64(2):129–37.CrossRefGoogle Scholar
  3. 3.
    Peyman GA, Charles HC, Liu KR, Khoobehi B, Niesman M. Intravitreal liposome-encapsulated drugs: a preliminary human report. Int Ophthalmol. 1988;12:175–82.CrossRefGoogle Scholar
  4. 4.
    Kishore K et al. Intravitreal clindamycin and dexamethasone for toxoplasmic retinochoroiditis. Ophthalmic Surg Lasers. 2001;32:183–92.PubMedGoogle Scholar
  5. 5.
    Sobrin L, Kump LI, Foster CS. Intravitreal clindamycin for toxoplasmic retinochoroiditis. Retina. 2007;27:952–7.CrossRefGoogle Scholar
  6. 6.
    Lasave AF, Díaz-Llopis M, Muccioli C, Belfort R, Arevalo JF. Intravitreal clindamycin and dexamethasone for zone 1 toxoplasmic retinochoroiditis at 24 months. Ophthalmology. 2010;117:1831–8.CrossRefGoogle Scholar
  7. 7.
    Soheilian M, Ramezani A, Azimzadeh A, Sadoughi MM, Dehghan MH, Shahghdami R, et al. Randomized trial of intravitreal clindamycin and dexamethasone versus pyrimethamine, sulfadiazine, and prednisolone in treatment of ocular toxoplasmosis. Ophthalmology. 2011;118:134–41.CrossRefGoogle Scholar
  8. 8.
    Tabbara KF, O’Connor GR. Treatment of ocular toxoplasmosis with clin- damycin and sulfadiazine. Ophthalmology. 1980;87:129–34.CrossRefGoogle Scholar
  9. 9.
    Maenz M, Schlüter D, Liesenfeld O, Schares G, Gross U, Pleyer A. Ocular toxoplasmosis past, present and new aspects of an old disease. Prog Retin Eye Res. 2014;39:77–106.CrossRefGoogle Scholar
  10. 10.
    Neu HC, Pnnce A, Neu CO, Garvey GJ. Incidence of diarrhea and colitis associated with clindamycim therapy. Infect Dis. 1977;135:120–5.CrossRefGoogle Scholar
  11. 11.
    Stainer GA, Peyman GA, Meisels H, Fishman G. Toxicity of selected antibiotics in vitreous replacement fluid. Ann Ophthalmol. 1977;9(5):615–8.PubMedGoogle Scholar
  12. 12.
    Walter P, Lüke C, Sickel W. Antibiotics and light responses in superfused bovine retina. Cell Mol Neurobiol. 1999;19(1):87–92.CrossRefGoogle Scholar
  13. 13.
    Tamaddon L, Mostafavi A, Riazi-esfahani M, Karkhane R, Aghazadeh S, Rafiee-Tehrani M, et al. Development, characterizations and biocompatibility evaluations of intravitreal lipid implants. Jundishapur J Nat Pharm Prod. 2014;9(2), e16414.CrossRefGoogle Scholar
  14. 14.
    Fernandes-Cunha GM, Gouvea DR, Fulgêncio GO, Rezende CMF, Da Silva GR, Bretas JM, et al. Development of a method to quantify clindamycin in vitreous humor of rabbits’ eyes by UPLC–MS/MS: application to a comparative pharmacokinetic study and in vivo ocular biocompatibility evaluation. J Pharm Biomed Anal. 2015;102:346–52.CrossRefGoogle Scholar
  15. 15.
    Fernandes-Cunha GM, Rezende CMF, Mussel WN, Da Silva GR, Gomes ECL, Yoshida MI, et al. Anti-toxoplasma activity and impact evaluation of lyophilization, hot molding process, and gamma-irradiation techniques on CLH-PLGA intravitreal implants. J Mater Sci Mater Med. 2016;27(10). doi: 10.1007/s10856-015-5621-1.
  16. 16.
    Fialho SL, Silva-Cunha A. Manufacturing techniques of biodegradable implants intended for intraocular application. Drug Deliv. 2005;12(2):109–16.CrossRefGoogle Scholar
  17. 17.
    Da Silva GR, Lima TH, Oréfice RL, Fernandes-Cunha GM, Silva-Cunha A, Zhao M, et al. In vitro and in vivo ocular biocompatibility of electrospun poly(e-caprolactone) nanofibers. Eur J Pharm Sci. 2015;73:9–19.CrossRefGoogle Scholar
  18. 18.
    Zhao M, Valamanesh F, Celerier I, Savoldelli M, Jonet L, Jeanny JC, et al. The neuroretina is a novel mineralocorticoid target: aldosterone up-regulates ion and water channels in Muller glial cells. FASEB J. 2010;24:3405–15.CrossRefGoogle Scholar
  19. 19.
    Zhao M, Célérier I, Bousquet E, Jeanny JC, Jonet L, Savoldelli M, et al. Mineralocorticoid receptor is involved in rat and human ocular chorioretinopathy. J Clin Invest. 2012;122(7):2672–9.CrossRefGoogle Scholar
  20. 20.
    Hezel M, Ebrahimi F, Kocha M, Dehghani F. Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012;43:1031–8.CrossRefGoogle Scholar
  21. 21.
    Brock WJ, Somps CJ, Torti V, Render JA, Jamison J, Rivera MI. Ocular toxicity assessment from systemically administered xenobiotics: considerations in drug development. Int J Toxicol. 2013;32(3):171–88.CrossRefGoogle Scholar
  22. 22.
    Söderstjerna E, Bauer P, Cedervall T, Abdshill H, Johansson F, Johansson UE. Silver and gold nanoparticles exposure to in vitro cultured retina—studies on nanoparticle internalization, apoptosis, oxidative stress, glial- and microglial activity. Plos One. 2014;9(8), e105359.CrossRefGoogle Scholar
  23. 23.
    Siqueira RC, dos Santos WF, Scott IU, Messias A, Rosa MN, Fernandes Cunha GM, et al. Neuroprotective effects of intravitreal triamcinolone acetonide and dexamethasone implant in rabbit retinas after pars plana vitrectomy and silicone oil injection. Retina. 2014;1–7.Google Scholar
  24. 24.
    Penha FM, Rodrigues EB, Maia M, Dib E, Fiod Costa E, Furlani BA, et al. Retinal and ocular toxicity in ocular application of drugs and chemicals—part I: animal models and toxicity assays. Ophthalmic Res. 2010;44(2):82–104.CrossRefGoogle Scholar
  25. 25.
    Bringmann A, Pannicke T, Grosche J, Francke M, Wiedemann P, Skatchkov SN, et al. Müller cells in the healthy and diseased retina. Prog Retin Eye Res. 2006;25:397–424.CrossRefGoogle Scholar
  26. 26.
    Chiba C. The retinal pigment epithelium: an important player of retinal disorders and regeneration. Exp Eye Res. 2014;123:107–14.CrossRefGoogle Scholar
  27. 27.
    Luo Y, Zhuo Y, Fukuhara M, Rizzolo LJ. Effects of culture conditions on heterogeneity and the apical junctional complex of the ARPE-19 cell line. Invest Ophthalmol Vis Sci. 2006;47(8):3644–55.CrossRefGoogle Scholar
  28. 28.
    Dubois-Dauphin M, Poitry-Yamate C, de Bilbao F, Julliard A, Jourdan F, Donati G. Early postnatal Müller cell death leads to retinal but not optic nerve degeneration in transgenic mice. Neuroscience. 2000;95:9–21.CrossRefGoogle Scholar
  29. 29.
    Ramadan GA. Sorbitol-induced diabetic-like retinal lesions in rats: microscopic study. Am J Pharmacol Toxicol. 2007;2(2):89–97.CrossRefGoogle Scholar
  30. 30.
    Maddala R, Reddy VN, Epstein DL, Rao V. Growth fator induced activation of Rho and RacGTPases and actin cytoskeletal reorganization in human lens epithelial cells. Mol Vis. 2003;9:329–36.PubMedGoogle Scholar
  31. 31.
    Hall A. Rho GTPases and the actin cytoskeleton. Science. 1998;279:509–14.CrossRefGoogle Scholar
  32. 32.
    Winkler J, Hagelstein S, Rohde M, Laqua H. Cellular and cytoskeletal dynamics within organ cultures of porcine neuroretina. Exp Eye Res. 2002;74:777–88.CrossRefGoogle Scholar
  33. 33.
    Verdugo-Gazdik ME, Simic D, Opsahl AC, Tengowski MW. Investigating cytoskeletal alterations as a potential marker of retinal and lens drug-related toxicity. Assay Drug Dev Technol. 2006;4(6):695–707.CrossRefGoogle Scholar
  34. 34.
    Ebert S, Schoeberl T, Walczak Y, Stoecker K, Stempfl T, Moehle C, et al. Chondroitin sulfate disaccharide stimulates microglia to adopt a novel regulatory phenotype. J Leukoc Biol. 2008;84(3):736–40.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Gabriella M. Fernandes-Cunha
    • 1
    • 2
    • 3
    • 4
    Email author
  • Silvia Ligório Fialho
    • 5
  • Gisele Rodrigues da Silva
    • 2
    • 3
    • 4
    • 6
  • Armando Silva-Cunha
    • 1
  • Min Zhao
    • 2
    • 3
    • 4
  • Francine Behar-Cohen
    • 2
    • 3
    • 4
  1. 1.Faculty of PharmacyFederal University of Minas GeraisBelo HorizonteBrazil
  2. 2.INSERM UMRS 1138, Team 17, Centre de Recherche des CordeliersParisFrance
  3. 3.Pierre and Marie Curie UniversityParisFrance
  4. 4.Paris Descartes UniversityParisFrance
  5. 5.Pharmaceutical and Biotechnological Development, Ezequiel Dias FoundationBelo HorizonteBrazil
  6. 6.Faculty of PharmacyFederal University of São João Del ReiDivinópolisBrazil

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