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1 wz (2009), 39 «2y J. Kor. Pharm. Sci., Vol. 39, No. 2, (2009) t y»» w»» w s sƒ ½x»Á½ ká w 1 Á x 1 Á½ 1 Á½ 2 Áx 2 Á û w w w, 1 t t»»», 2 û w œ w ( Á Á ) Cytotoxicity Evaluation on Hydrogels for Medical Devices based on the International Organization for Standardization Hyun-Ki Kim, Ye-Tae Kim, Yang Ha Cho 1, Hye-Won Roh 1, Min-A Kim 1, So Yeon Kim 2, Kang Moo Huh 2 and Jeong-Sook Park College of Pharmacy, Chungnam National University, 1 Medical Device Standardization Division, Korea Food and Drug Administration, 2 College of Engineering, Chungnam National University (Received March 25, 2009ÁRevised March 31, 2009ÁAccepted April 1, 2009) ABSTRACT Hydrogels for medical devices such as hydrophilic dressing, moisturizing healing band, hydrophilic intravenous catheter and soft contact lens were evaluated for their cytotoxicity according to the International Organization for Standardization (ISO) procedures. To test indirect cytotoxicity of hydrogel products, dissolution medium and dissolution condition were selected based on the guideline for medical devices. Cytotoxicity was low in all the case of hydrogel products. Soft contact lens showed no significant difference in dissolution between complete medium and saline. Currently, there is no specific guidelineto test hydrogel for medical devices in Korea with consideration of characteristics of hydrogel. Thus, proper method of cytotoxicity evaluation should be selected depending on the characteristics and usages of hydrogels for medical devices. Key words Hydrogel, International Organization for Standardization, Medical devices, Cytotoxicity w e š ƒ y, w w p. w q(slab)x, (microparticles), ù (nanoparticles), gq, v w xk w, w w w k w ùkü» ƒ Ÿ w x š. w w 1,2) ƒ», e», ell», 3), œv,, gkp»¾ w. w w 4-6) w w»» w w w t š,» v y w w. x w»» w w Tel : 042) , w p w ³»»» x w w w.»» x 7)»sƒ x s x, x, x (vü x), x, x, x, x, x w x, ƒ s ƒ x x, x, x, w x. 8) t y» (International Organization for Standardization, ISO)»» s s, xk, s y d t w ³ wš. ù 9) xwš w, s, x. 10) ƒ ww s x w sƒw» w.»» sƒw s x j x ù. 11) š»» s sd ew w w 127

2 128 ½x»Á½ ká wá x Á½ Á½ Áx Á w w ùkü, s v w w» y w s sƒw». x x» w w š w w, x sww, x sww,,»k w wù kw 37 C 24 w t y» (ISO) ³ w x w Table I. 12) ù, x t t š»» w ³ wš,»» w w ³ t y» ³ w x w s x ww»» w x w» wš w. x»»»» w x q t e (Hydrogel-Absorber F, ( ), w ), ( e v, ( ), w ), x ü p elk(angiocath Plus TM, m k g ( ), w ), vpgkp (Focus Dailies F, Ciba Vision GmbH, ) w w. s sƒ w w fetal bovine serum(fbs) Dulbecco`s modified Eagle Medium(DMEM) Gibco BRL(NY, ), š 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolum bromide (MTT) Sigma Aldrich Co.(St. Louis, MO, ) w w.»k q t w. s w s d w» w s HepG2 s(korean Cell Line Bank,, w ) w. s 10% FBS, penicillin U/ ml(gibco BRL), streptomycin 10 mg/ml(gibco BRL) w w DMEM w plastic culture dish 37 C, 5% CO 2 24 w. t»» w w w w yw y Table I Suitable Extraction Ratios for Test Materials of Various Thicknesses 12) Thickness (mm) Extraction ratio (surface area/ volume)±10% < cm 2 /ml Examples of material Film, sheet, tubing wall cm 2 /ml Tubing wall, slab, small molded items > cm 2 /ml Large molded item(s) Irregular shape of solid medical devices Irregular shape of porous medical devices (low density of raw material) 0.2 g/ml Powder, pellet, foam, non-absorbent, molded items 0.1 g/ml Membrane sƒw» w s x ww» w w w. ww» UV 15 w, ³ k w, ³ k ww w s w š w. 13) s w w x w w w š, vpgkp w» x w w w. ISO ³ ƒ t w Ì Digimatic caliper (Mitutoyo Co., ) d w Table I l w, 12) 37±2 C 48 k ww. s x w s sƒw» w MTT assay w. HepG2 s plate w w z s d w 96-well plate cells/well s wš w w. 13) sƒ z wš w w 200 µl 96 well ƒw. 37 C 24 z, wš ƒ well 100 µl MTT (5 mg/ml) ƒw. 37 C 4 k z MTT wš, ƒ well MTT solubilization solution 100 µl ƒw. 30 ew z Microplate reader (Sunrise, Tecan Trading, ) 570 nm Ÿ d w, ƒ x 4z ww. Ÿ d w

3 t y»» w»» w s sƒ 129 Figure 1 Cytotoxicity of hydrogel for hydrophilic dressing product (a) and moisturizing healing band (b) by ISO method. Each value represents the mean±s.d. (n=4). z w s w s q w. s (%) = (Abs test /Abs control ) 100 m l s³ ±t r (s.d.) ùkü, vpgkp s t- test w p<0.05 š q w. š t y» x w w, x w w,,»k w wù w «w. w w t ù x x w w w. w 37±2 C 24, 50±2 C 72±2, 70±2 C 24±2, 121±2 C 1±0.2 ƒ w, x sw w 37±2 C 24 w x 37±2 C 48 k ww. e ̃ ƒƒ 0.44 mm 0.33 mm, Table I l Ì xk š w 6cm /ml 2 w. e w k, s (610 mg/ml) 2, 4, 8, 16, 32 HepG2 s ƒƒ 57.7%, 61.4%, 64.5%, 72.0%, 81.4%, 83.2% s ùkü (Figure 1a). w k s (680 mg/ml) 2, 4, 8, 16, 32 HepG2 s ƒw ƒƒ 35.5%, 54.4%, 73.7%, 87.9%, 91.0%, 92.3% s ù kü (Figure 1b). 2, 4 s û ùkû. x e 610 mg/ml 680 mg/ml, s x 32 ƒƒ 19 mg/ml, 21 mg/ml. Geever 14) w š 25 mg/ml ƒ¾ š š w w s û. e x ell ̃ 0.5 mm w š, p x Table I w 6cm /ml 2 w. x ell x w t wš w v ƒw w. x ell w k, s 2, 4, 8, 16, 32 HepG2 s s ƒƒ 83.2%, 87.1%, 89.4%, 92.9%, 100.7%, 101.8% s ùkü (Figure 2).» p 4g 20 ml w ù,» ell 2g 20 ml 24 w s xw û ùk û. x 48 w ù 15) s ùkù. vpgkp xk š ̃ 0.5 mm w, Table I w 6cm /ml 2 w. vpgkp

4 130 ½x»Á½ ká wá x Á½ Á½ Áx Á Figure 2 Cytotoxicity of hydrogel for hydrophilic intravenous catheter by ISO method. Each value represents the mean±s.d. (n=4). Figure 3 Cytotoxicity of hydrogel for soft contact lens by ISO method. Each value represents the mean±s.d. (n=4). *p<0.05, saline as a dissolution medium compared to complete medium.» x w w w, w» ƒ w. vpgkp w s, 2, 4, 8, 16, 32 ƒw ƒƒ 46.4%, 74.8%, 80.4%, 89.2%, 93.8%, 94.4% s ùkü (Figure 3). vpgkp k 2, 4, 8, 16, 32 s ƒw ƒƒ 28.4%, 73.0%, 76.1%, 82.5%, 85.4%, 86.3% s ùkü (Figure 3). w,, 8, 32 s ƒw (p<0.05) ùkü. ù, 8, 32 80% s ùkü š, s v w ƒ sw» û s ùkü w». vpgkp s ùkü» x s û sƒ. x, ù w g w t y x»» sƒ wš š. ƒ w, d w 2g x š, vƒ 2g 20 mlƒ w «wš. 12)»» s x»» w x w t w s ùkû. w, vpgkp x w w s ùkü ù w w. x»» sƒ w»»» w ƒ wù, w w p š w w» w v w. t t (08142»372) w w. š x 1) T. R. Hoare and D. S. Kohane, Hydrogels in drug delivery: Progress and challenges, Polymer, 49, (2008). 2) S. S. Oh, C. J. Kim, H. S. Kim and Y. H. Shin, Efficacy of hydrogel patch in wound rat model, J. Kor. Pharm. Sci., 39, (2009). 3) R. J. Whitbourne, Lubricious hydrophilic coating, resistant to wet abrasion. Patent no. 5,331,027 (1994). 4) J. Varshosaz and N. Koopaie, Cross-linked poly(vinyl alcohol) hydrogel: study of swelling and drug release behaviour, Iran. Polym. J., 11, (2002). 5) A. Akhgari, H. Afrasiabi Garekani, F. Sadeghi and M.

5 t y»» w»» w s sƒ 131 Azimaie, Statical optimization of indomethacin pettets coated with ph-dependent methacrylic polymers for possible colonic drug delivery, Int. J. Pharm., 305, (2005). 6) D. M. Devine, S. M. Devery, J. G. Lyons, L. M. Geever, J. E. Kennedy and C. L. Higginbotham, Multifunctional polyvinylpyrrolidone-polacrylic acid copolymer hydrogels for biomedical applications, Int. J. Pharm., 326, (2006). 7) t t š y ( ),»» w w œm» ³. 8) t t»»,»» w sƒ ƒ (2008). 9) G. Eisenbrand, B. Pool-Zobel, V. Baker, M. Balls, B. J. Blaauboer, A. Boobis, A. Carere, S. Kevekordes, J. C. Lhuguenot, R. Pieters and J. Kleiner, Method of in vitro toxicology, Food Chem. Toxicol., 40, (2002). 10) J. Weyermann, D. Lochmann and A. Zimmer, A practical note on the use of cytotoxicity assays, Int. J. Pharm., 288, (2005). 11) ISO (International Organization for Standardization) Biological evaluation of medical devices. Part 5: tests for in vitro cytotoxicity (1999). 12) ISO (International Organization for Standardization) Biological evaluation of medical devices. Part 12: sample preparation and reference materials (2007). 13) G. V. N. Rathna and Gelatin hydrogels: enhanced biocompatibility, drug release and cell viability, J. Mater. Sci. Mater. Med., 19, (2008). 14) L. M. Geever, C. C. Cooney, J. G. Lyons, J. E. Kennedy, M. J. D. Nugent, S. Devery and C. L. Higginbotham, Characterisation and controlled drug release from novel drug-loaded hydrogels, Eur. J. Pharm. Biopharm., 69, (2008). 15) G. Chaiban, H. Hanna, T. Dvorak and I. Raad, A rapid method of impregnating endotracheal tubes and urinary catheters with gendine: a novel antiseptic agent, J. Antimicrob. Chemother., 55, (2005).

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