Effect of Fullerenol C60 on Erythrocyte Deformability During Ischaemia-Reperfusion Injury of Lower Extremity in Diabetic Rats

  • Gülay Kip
  • Hakan Kartal
  • Faruk Metin Çomu
  • Yücel Polat Gazi University, Faculty of Medicine, Department of Anaesthesiology and Reanimation, Ankara, Turkey
  • Mustafa Arslan
  • Ayşegül Küçük


Background: Fullerenol, a water-soluble C60-fullerene derivative synthesized by Chiang et al, has been demonstrated to be able to scavenge free radicals in vitro and in vivo. Although its protective effects have been already studied and shown in ischemia reperfusion (IR) injury, additional investigation is necessary for its effect on erythrocyte deformability. The purpose of our study was to look into the effects of fullerenol C60 on erythrocyte deformability in rat lower extremity ischemia reperfusion injury model. Materials and Methods: After approval of the Ethics Committee,  30 Wistar Albino rat were divided into 5 groups (n:6) as; Control (C), Diabetes (group D), diabetes+ fullerenol C60 group (DF), diabetes+ IR (group DIR) and diabetes IR+ fullerenol C60 (DIRF). 55 mg/kg streptozotocin was administered to the rats for diabetes. After the period of 72 hour,  blood glucose concentration was mesured, 250 mg/dl and above were considered as diabetic rat. Four week after the formation of diabetes, rats were subjected to 2 hour ischemia and 2 hour reperfusion. Erythrocyte packs  were prepared from heparinized blood samples and deformability measurements were performed. Results: The deformability index was significantly increased in diabetic rats; however, it was similar in group D, DF and DIRF. It was significantly increased in group DIR when compared to group C, D, DF and DIRF. The relative resistance was increased in I/R models. Conclusion: This study aimed to investigate the effects of IR on erythrocyte deformability which may lead to disturbance in blood flow and hence tissue perfusion in infrarenal rat aorta. We found that fullerenol C60 had beneficial effects by reversing undesirable effects of IR. In our opinion, further studies with larger volume are required to support our promising results.


Duru S, Koca U, Oztekin S, Olguner C, Kar A, Coker C, et al. Antithrombin III pretreatment reduces neutrophil recruitment into the lung and skeletal muscle tissues in the rat model of bilateral lower limb and reperfusion: A pilot study. Acta Anaesthesiol Scand 2005;49:1142-8.

Turchányi B, Tóth B, Rácz I, Vendégh Z, Furész J, Hamar J. Ischemia reperfusion injury of skeletal muscle after selective deafferantation. Physiol Res 2005;54:25-32.

Lin B, Ginsberg M, Busto R, Li L. Hyperglycemia triggers massive neutrophil deposition in brain following transient ischemia in rats. Neurosci Lett 2000;278:1-4.

Walters TJ, Garg K, Corona BT. Activity attenuates skeletal muscle fiber damage after ischemia and reperfusion. Muscle Nerve 2015;52:640-8.

Prylutskyy YuI, Durov SS, Bulavin LA, Adamenko II, Moroz KO, Geru II, et al. Structure and thermophysical properties of fullerene C60 aqueous solutions. Int J Thermophys 2001;22:943-56.

Scharff P, Carta-Abelmann L, Siegmund C, Matyshevska OP, Prylutska SV, Koval TV, et al. Effect of X-ray and UV irradiation of the C60 fullerene aqueous solution on biological samples. Carbon 2004;42:1199-201.

Prylutska SV, Matyshevska OP, Grynyuk II, Prylutskyy YuI, Ritter U, Scharff P. Biological effects of C60 fullerenes in vitro and in a model system. Mol Cryst Liq Cryst 2007;468:265-74.

Prylutska SV, Burlaka AP, Prylutskyy YI, Ritter U, Scharff P. Pristine C60 fullerenes inhibit the rate of tumor growth and metastasis. Exp Oncol 2011;33:162-4.

Zay SY, Zavodovsky DA, Bogutska KI, Nozdrenko DN, Prylutskyy YI. Prospects of C60 fullerene application as a mean of prevention and correction of ischemic-reperfusion injury in the skeletal muscle tissue. Fiziol Zh 2016;62:66-77.

Prylutska SV, Matyshevska OP, Golub AA, Prylutskyy YuI, Potebnya GP, Ritter U, et al. Study of C60 fullerenes and C60-containing composites cytotoxicity in vitro. Mater Sci Engineer C 2007;27:1121-24.

Prylutska SV, Grynyuk II, Grebinyk SM, Matyshevska OP, Prylutskyy YuI, Ritter U, et al. Comparative study of biological action of fullerenes C60 and carbon nanotubes in thymus cells. Mat-wiss u Werkstofftech 2009;40:238-41.

Tolkachov M, Sokolova V, Korolovych V, Prylutskyy Yu, Epple M, Ritter U, et al. Study of biocompatibility effect of nanocarbon particles on various cell types in vitro. Mat-wiss u Werkstofftech 2016;47:216-21.

Burlaka AP, Sidorik YP, Prylutska SV, Matyshevska OP, Golub OA, Prylutskyy YI, et al. Catalytic system of the reactive oxygen species on the C60 fullerene basis. Exp Oncol 2004;6:326-7.

Prylutska SV, Grynyuk II, Matyshevska OP, Prylutskyy YuI, Ritter U, Scharff P. Anti-oxidant properties of C60 fullerenes in vitro. Fullerenes Nanotubes Carbon Nanostruct 2008;16:698-705.

Gharbi N, Pressac M, Hadchouel M, Szwarc H, Wilson SR, Moussa F. C60 fullerene is a powerful antioxidant in vivo with no acute or subacute toxicity. Nano Lett 2005;5:2578-85.

Milic VD, Stankov K, Injac R, Djordjevic A, Srdjenovic B, Govedarica B, et al. Activity of Antioxidative Enzymes in Erythrocytes after a Single Dose Administration of Doxorubicin in Rats Pretreated with Fullerenol C60(OH)24. Toxicol Mech Methods 2009;19:24-8.

Injac R, Perse M, Cerne M, Potocnik N, Radic N, Govedarica B, et al. Protective effects of fullerenol C60(OH)24 against doxorubicin-induced cardiotoxicity and hepatotoxicity in rats with colorectal cancer. Biomaterials 2009;30:1184-96.

Chen C, Xing G, Wang J, Zhao Y, Li B, Tang J, Jia G, et al. Multihydroxylated [Gd@C82(OH)22]n nanoparticles: antineoplastic activity of high efficiency and low toxicity. Nano Lett 2005;5:2050-7.

Nozdrenko DM, Zavodovskyi DO, Matvienko TY, Zay SY, Bogutska KI, Prylutskyy YI et al. C60 Fullerene as Promising Therapeutic Agent for the Prevention and Correction of Skeletal Muscle Functioning at Ischemic Injury. Nanoscale Res Lett 2017;12:115. doi: 10.1186/s11671-017-1876-4.

Thompson LC, Urankar RN, Holland NA, Vidanapathirana AK, Pitzer JE, Han L, et al. C₆₀ exposure augments cardiac ischemia/reperfusion injury and coronary artery contraction in Sprague Dawley rats. Toxicol Sci 2014;138:365-78.

Montalvo-Jave EE, Escalante-Tattersfield T, Ortega-Salgado JA, Pina E, Geller DA. Factors in the pathophysiology of the liver ischemia-reperfusion injury. J Surg Res 2008;147:153-9.

Xu Z, Yu J, Wu J, Qi F, Wang H, Wang Z, et al. The effects of two anesthetics, propofol and sevoflurane, on liver ischemia/reperfusion injury. Cell Physiol Biochem 2016;38:1631-42.

Grebowski J, Krokosz A, Puchala M. Membrane fluidity and activity of membrane ATPases in human erythrocytes under the influence of polyhydroxylated fullerene. Biochim Biophys Acta 2013;1828:241-8.

Grebowski J, Krokosz A, Puchala M. Fullerenol C60(OH)36 could associate to band 3 protein of human erythrocyte membranes. Biochim Biophys Acta. 2013;1828:2007-14.

Grebowski J, Krokosz A. The Effect of Highly Hydroxylated Fullerenol C60(OH)36 on Human Erythrocyte Membrane Organization. Journal of Spectroscopy 2015;10.1155/2015/825914.

Chiang LY, Lu FJ, Lin JT. Free radical scavenging activity of water-soluble fullerenols. J Chem Soc Chem Commun 1995;12:1283-4.

Nielsen GD, Roursgaard M, Jensen KA, Poulsen SS, Larsen ST. In vivo biology and toxicology of fullerenes and their derivatives. Basic Clin Pharmacol Toxicol 2008;103:197-208.

Markovic Z, Trajkovic V. Biomedical potential of the reactive oxygen species generation and quenching by fullerenes (C60). Biomaterials 2008;29:3561-73.

Mirkov SM, Djordjevic AN, Andric NL, Andric SA, Kostic TS, Bogdanovic GM, et al. Nitric oxide-scavenging activity of polyhydroxylated fullerenol, C60(OH)24. Nitric Oxide 2004;11:201-7.

Zinchuk VV. Erythrocyte deformability: physiological aspects. Usp Fiziol Nauk 2001;32(3):66-78.

Kuypers FA. Red cell membrane damage. J Heart Valve Dis 1998;7:387-95.

Sivilotti ML. Oxidant stres and haemolysis of the human erythrocyte. Toxicol Rev 2004;23:169-88.

Grebowski J, Kazmierska P, Litwinienko G, Lankoff A, Wolszczak M, Krokosz A. Fullerenol C60(OH)36 protects human erythrocyte membrane against high-energy electrons. Biochim Biophys Acta Biomembr 2018;1860:1528-36.

Original Research