In vitro amoebicidal activity of poly(maleic anhydride-co-vinyl acetate) copolymer on Acanthamoeba spp. trophozoites and cysts

Abstract

Abstract

Aim. Poly(maleic anhydride-co-vinyl acetate) copolymer (MAVA) has been known to confer antitumor activity and it also exhibit many other biological activities. The amoebicidal effect of MAVA has not been investigated. Methods. In this study, for the first time amoebicidal effect of MAVA was investigated on Acanthamoeba spp. trophozoites and cysts. Trypan Blue Dye Exclusion test was used to determine the cell viability. Results. Our findings indicated that in the period of 48 h, the percentage and number of viable trophozoite showed a slow decrease except the dose of 32.0 mg/mL. The copolymer in the highest dose inhibited the proliferation of trophozoites in 3 h. The cysts were more resistant than the trophozoites to the inhibitory effect of copolymer. Structural characterization of the copolymer was confirmed by Fourier Transform Infrared (FTIR) and Nuclear Magnetic Resonance (¹H-NMR). Surface morphology was visualized by both scanning electron and atomic force microscopy. Conclusion. In the lights of these data it could be suggested that MAVA deserves deeper investigation to improve its amoebicidal activity for both trophozoites and cysts forms of Acanthamoeba spp.

Keywords: poly(maleic anhydride-co-vinyl acetate) copolymer; Acanthamoeba spp. trophozoite; Acanthamoeba sp. cysts; amoebicidal activity; bioactive polymers.

Özet

Amaç. Poli(maleik anhidrit-ko-vinil asetat) kopolimerinin (MAVA) antitümoral aktiviteye sahip olduğu bilinmektedir ve aynı zamanda çeşitli biyolojik aktivitelere sahiptir. MAVA'nın amobisidal etkisi daha önce incelenmemiştir. Yöntem. Bu araştırmada ilk kez MAVA'nın amobisidal aktivitesi Acanthamoeba trofozoit ve kistleri üzerinde incelendi. Hücre canlılığını belirlemek için Tripan mavisi boyası kullanıldı. Bulgular. Bulgularımız ilk 48 saatte 32,0 mg/mL dozu dışında canlı trofozoit oranı ve sayısının hızlı bir azalma gösterdiğini ortaya koydu. MAVA en yüksek dozunda 3 saat içinde trofozoitlerin çoğalmasını baskıladı. Kistlerin bu baskılayıcı etkiye daha dirençli oldukları saptandı. MAVA'nın yapısal karakterizasyonu Fourier Transform Infrared ve nükleer manyetik rezonans (¹H-NMR) ile doğrulandı. Yüzey morfolojisi ise hem taramalı elektron hem de atomik kuvvet mikroskobu ile görüntülendi. Sonuç. Bulgularımızın ışığında MAVA, Acanthamoeba trofozoit ve kistleri üzerinde gösterdiği amobisidal etkilerden dolayı daha fazla araştırılmayı hak etmektedir.

Anahtar sözcükler: Poli(maleik anhidrit-ko-vinil asetat) kopolimeri, Acanthamoeba spp. trofozoiti, Acanthamoeba spp. kisti, amobisidal aktivite, biyoaktif polimerler

Keywords

• poly(maleic anhydride-co-vinyl acetate) copolymer; Acanthamoeba spp. trophozoite; Acanthamoeba sp. cysts; amoebicidal activity; bioactive polymers.

-

Abstract

Amaç. Poli(maleik anhidrit-ko-vinil asetat) kopolimerinin (MAVA) antitümoral aktiviteye sahip olduğu bilinmektedir ve aynı zamanda çeşitli biyolojik aktivitelere Corresponding Author: Dr. Gülderen Karakuş, CÜTFAM, Cumhuriyet Üniversitesi Tıp Fakültesi, TR-58140 Sivas. Email: [e-mail] sahiptir. MAVA’nın amobisidal etkisi daha önce incelenmemiştir. Yöntem. Bu araştırmada ilk kez MAVA’nın amobisidal aktivitesi Acanthamoeba trofozoit ve kistleri üzerinde incelendi. Hücre canlılığını belirlemek için Tripan mavisi boyası kullanıldı. Bulgular. Bulgularımız ilk 48 saatte 32,0 mg/mL dozu dışında canlı trofozoit oranı ve sayısının hızlı bir azalma gösterdiğini ortaya koydu. MAVA en yüksek dozunda 3 saat içinde trofozoitlerin çoğalmasını baskıladı. Kistlerin bu baskılayıcı etkiye daha dirençli oldukları saptandı. MAVA’nın yapısal karakterizasyonu Fourier Transform Infrared ve nükleer manyetik rezonans (H-NMR) ile doğrulandı. Yüzey morfolojisi ise hem taramalı elektron hem de atomik kuvvet mikroskobu ile görüntülendi. Sonuç. Bulgularımızın ışığında MAVA, Acanthamoeba trofozoit ve kistleri üzerinde gösterdiği amobisidal etkilerden dolayı daha fazla araştırılmayı hak etmektedir.

Keywords

• Poli(maleik anhidrit-ko-vinil asetat) kopolimeri, Acanthamoeba spp. trofozoiti, Acanthamoeba spp. kisti, amobisidal aktivite, biyoaktif polimerler

References

1. Felthouse TR, Burnett JC, Horrell B, Mummey MJ, Kuo YJ. Chemical Technology 2001, DOI: 10.1002/0471238961.1301120506051220, Huntsman Petrochemical Corporation Austin Laboratories 7114 North Lamar Boulevard Austin, Texas, pp 1
2. Spridon D, Panaitescu L, Ursu D, and Uglea CV. Synthesis and biocompatibility of maleic anhydride copolymers: 1. maleic anhydride-vinyl acetate, maleic anhydride methyl methacrylate and maleic anhydride-styrene. Polym Int. 1997;43:175-81.
3. Breslow DS. Biologically active synthetic polymers. Pure &Appl Chem. 1976; 46: 103Dhal PK, Holmes-Farley SR, Huval CC, and Jozefiak TH. Polymers as drugs. AdvPolym Sci. 2006;192:9-58.
4. Duncan R. The dawning era of polymer therapeutics. Nat Rev Drug Discov. 2003 May;2(5):347-60.
5. Martinez AJ, Visvesvara GS. Free-living, amphizoic and opportunistic amabas. Brain Pathol. 1997 Jan;7(1):583-598.
6. Değerli S, Saygı G. Acanthamoebakeratiti. TurkiyeParazitolDerg. 2000;24:243-8.
7. Claerhout I, Kestelyn PH. Acanthamoeba keratitis: a review. Bull SocOphtalmol. 1999;274,71-82.
8. Walia R, Montoya JG, Visvesvera GS, Booton GC, Doyle RL. A case of successful treatment of cutaneous Acanthamoeba infection in a lung transplant recipient. Transpl Infect Dis. 2007 Mar;9(1), 51-4.

9. MacLean RC, Hafez N, Tripathi S, Childress CG, Ghatak NR, Marciano-Cabral F. Identification of Acanthamoeba sp. İn parafin-embedded CNS tissue from an HIV+ individual by PCR. DiagnMicrobiol Infect Dis. 2007 Mar; 57(3):289-94.
10. Rivera M, Padhya T. Acanthamoeba: a rare primary cause of chinosinusitis. Laryngoscope. 2002 Jul;112(7 Pt 1):1201-3.
11. Cabral FM, Cabral G. Acanthamoeba spp. As agents of disease in humans. ClinMicrobiol Rev. 2003 Apr;16(2), 273-307.
12. Pasricha G, Sharma S, Garg P, Aggarwal RK. Use of 18S rRNA gene-based PCR assay for diagnosis of Acanthamoeba keratitis in non-contact lens wearers in India. J ClinMicrobiol. 2003 Jul; 41(7):3206-11.
13. Sun X, Zhang Y, Li R, Wang Z, Luo S, Gao M, Deng S, Chen W, Jin X. Acanthamoeba keratitis: clinical characteristics and management. Ophthalmology, 2006 Mar;113(3), 412–6.
14. Dart JK, Saw VP, Kilvington S. Acanthamoeba keratitis: diagnosis and treatment update 2009. Am J Ophthalmol. 2009 Oct;148(4):487-499.
15. Ertabaklar H, Dayanir V, Apaydin P, Ertug S, Walochnik J. Case report: Acanthamoeba keratitis. TurkiyeParazitolDerg. 2009;33(4):283-5.
16. Singhal T, Bajpai A, Kalra V, Kabra SK, Samantaray JK, Satpathy G, Gupta AK. Pediatr Infect Dis J. 2001 Jun; 20(6), 623–7.
17. Yoon KJ, Woo JH, Seo YS. Formaldehyde free cross-linking agents based on maleic anhydride copolymers. Fiber Polym. 2003;4:182-187.
18. Nemtoi G, Beldie C, Tircolea C, Popa I, Cretescu I, Humelnicu I, Humelnicu D. Behaviour of the poly(maleic anhydride-co-vinyl acetate) copolymer in aqueous solutions. EurPolym J. 2001;37:729-35.
19. Xing CM, Yang WT. A novel, facile method for the preparation of uniform, reactive maleic anhydride/vinyl acetate copolymer micro- and nanospheres. Macromol Rapid Comm. 2004;25:1568-74.
20. Nguyen V, Yoshida W, Cohen Y. Graft polymerization of vinyl acetate onto silica. J ApplPolym Sci. 2003;87:300-310.
21. Karakus G, Zengin HB, Polat ZA, Yenidunya AF, Aydin S. Cytotoxicity of three maleic anhydride copolymers and common solvents used for polymer solvation. Polym Bull. 2012; DOI: 10.1007/s00289-012-0860-5.
22. Schuster FL. Cultivation of pathogenic and opportunistic free-living amebas. ClinMicrobiol Rev. 2002 Jul; 15(3):342-54.
23. Garcia LS, Brucker DA, Diagnostic medical parasitology, 2nd Ed., American Society for Microbiology, Washington, pp 601 (1993).
24. Khan NA.Acanthamoeba Biology and Pathogenesis, Caister Academic Press, Norfolk, UK, pp 85 (2009).
25. Ballarin C, Peruffo A. Primary cultures of astrocytes from fetal bovine brain. Methods in Molecular Biology. 2012; 814:117-26.
26. Rendules O, Ferrières L, Frètaud M, Bègaud E, Herbomel P, Levraud JP, Ghigo JM. A new zebrafish model of oro-intestinal pathogen colonization reveals a key role for adhesion in protection by probiotic bacteria. Plos Pathogens. 2012;8(7);1-17.
27. Kaplan Can H, Doğan AL, Rzaev ZMO, Uner, AH, Güner A. Synthesis and antitumor activity of poly(3,4-dihydro-2H-pyran-co-maleic anhydride-co-vinyl acetate). J ApplPolym Sci. 2005;96:2352-59.
28. Pal J, Singh H, Ghosh AK. Modification of LLDPE using esterified styrene maleic anhydride copolymer: Study of its properties and environmental degradability. J ApplPolym Sci. 2004;92:102-8.
29. Popa I, Offenberg H, Beldie C, Uglea CV. Benzocaine modified maleic anhydride copolymers-I. Synthesis and characterization of benzocaine modified poly(maleic anhydride-co-vinyl acetate), poly(maleic anhydride-co-methyl methacrylate) and poly(maleic anhydride-co-styrene). EurPolym J. 1997;33:1511-14.
30. Qiao Z, Xie Y, Chen M, Xu J, Zhu Y, Qian Y. Synthesis of lead sulfide/(polyvinyl acetate) nanocomposites with controllable morphology. ChemPhysLett. 2000;321:504-7.
31. Giessibl F. Advances in Atomic Force Microscopy. J Rev Mod Phys. 2003;75: 949
32. Cowman MK, Li M, Balazs EA. Tapping mode atomic microscopy of hyaluronan: extended and intramolecularly interacting chains. Biophys J. 1998 Oct;75(4), 2030
33. Hernández JCR, Sánchez MS, Ribelles JLG, Pradas MM. Polymer-silica nanocomposites prepared by sol-gel technique: Nanoindentation and tapping mode AFM. EurPolym J. 2007;43:2775-2783.
34. Kuyyakanond T, Quesnel LB. The mechanism of action of chlohexidine. FEMS MicrobiolLett. 1992 Dec 15;79(1-3):211–5.
35. El-Sayed NM, Ismail KA, Ahmed SA, Hetta MH. In vitro amoebicidal activity of ethanol extracts of Arachis hypogaea L., Curcuma longa L. and Pancratium maritimum L. on Acanthamoeba castellanii cysts. Parasitol Res. 2012 May; 110(5):1985-92.
36. Allen PG, Dawidowicz EA. Phagocytosis in Acanthamoeba: I. A mannose receptor is responsible fort he binding and phagocytosis of yeast. J Cell Physiol. 1990 Dec; 145(3):508-13.
37. Roberts CW, Henriquez FL. Drug target identification, validation, characterisation and exploitation for treatment of Acanthamoeba (species) infections. ExpParasitol. 2010 Sep; 126(1):91–6.
38. Cariello AJ, Souza GFP, Foronda AS, Yu MCZ, Hofling-Lima AL, Ganzarolli M. In vitro amoebicidal activity of S-nitrosoglutathione and S-nitroso-N-acetylcysteine against trophozoites of Acanthamoebacastellanii. J AntimicrobChemother. 2010 Mar; 65(3), 588-91.
39. Alizadeh H, Neelam S, Cavanagh HD. Amoebicidal activities of alexidine against 3 pathogenic strains of acanthamoeba. Eye Contact Lens. 2009 Jan; 35(1):1-5.
40. Goze I, Ali̇m A, Dag S, Tepe B, Polat ZA. In vitro amoebicidal activity of Salvia staminea and Salvia caespitosa on Acanthamoeba castellanii and their cytotoxic potentials on corneal cells. J OculPharmacolTher. 2009 Aug; 25(4), 293-8.
41. Topalkara A, Vural A, Polat Z, Toker MI, Arici MK, Ozan F, Cetin A. In vitro amoebicidal activity of propolis on Acanthamoebacastellanii. J OculPharmacolTher. 2007 Feb; 23(1):40-55.
42. Malatyali E, Tepe B, Degerli S, Berk S, Akpulat HA. In vitro amoebicidal activity of four Peucedanum species on Acanthamoebacastellanii cysts and trophozoites. Parasitol Res. 2012 Jan; 110(1):167-74.
43. Akin Polat Z, Vural A, Tepe B, Cetin A. In vitro amoebicidal activity of four Allium species on Acanthamoebacastellanii and their cytotoxic potentials on corneal cells. Parasitol Res. 2007 Jul; 101(2), 397-402.
44. Akin Polat Z, Tepe B, Vural A. In vitro effectiveness of Thymus sipyleussubsp.Sipyleusvar.Sipyleus on Acanthamoebacastellanii and its cytotoxic potential on corneal cells. Parasitol Res. 2007 Nov; 101(6), 1551-5.