Behavior of Hepatocytes Inoculated in Gelatin-Immobilized Polyurethane Foam

Kyung-Su Yang; Xinglin Guo; Wan Meng; Jae-Yong Hyun; Inn-Kyu Kang; Yang Il Kim

Macromolecular Research, Vol.11, No.6, 488-494, December, 2003

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Behavior of Hepatocytes Inoculated in Gelatin-Immobilized Polyurethane Foam

Kyung-Su Yang, Xinglin Guo, Wan Meng, Jae-Yong Hyun, Inn-Kyu Kang^†, and Yang Il Kim¹

Department of Polymer Science, Kyungpook National University, Daegu 702-701, Korea
¹School of Medicine, Kyungpook National University, Daegu 700-091, Korea

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We have fabricated gelatin-immobilized polyurethane foams (PUFG) by dipping polyurethane foam (PUF) in an aqueous solution containing gelatin and by subsequent reaction with glutaraldehyde after freeze-drying. Gelatin aqueous solutions of different concentrations were used as the dipping solutions to control the amount of immobilized gelatin. The average pore size of PUF decreased with an increase in gelatin concentration. It was found from the hepatocyte adhesion experiment that the amount of hepatocytes seeded on PUFG1, prepared by using a 1% aqueous gelatin solution, was higher than that on other PUFGs. The hepatocytes inoculated in PUFG1 were slightly aggregated as the incubation time increased. The cells inoculated in PUFG1 showed higher ammonia removal ability than those monolayer-cultured on a gelatin-immobilized polystyrene dish (PSG) after 1 and 4 days of incubation time. The inoculated cells exhibited higher albumin secretion relative to monolayer-cultured hepatocytes on PSG. Albumin secretion by hepatocytes seeded on PUFG1 was increased by the presence of serum and was further increased by both the presence of serum and cytokines. The results obtained from a 3-(3,4-dimethylthiazol-2yl)-2,5- diphenyltetrazolium bromide (MTT) assay indicated that PUFG can provide a better microenvironment for hepatocyte culture along with nutrition and metabolite transfer through the high porosity of PUF.

Keywords:gelatin;polyurethane foam;hepatocyte;albumin secretion.

[References]

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Fukuda J, Sakiyama R, Nakazawa K, Ijima H, Yamashita Y, Shimada M, Shirabe K, Tsujita E, Sugimachi K, Funatsu K, Int. J. Artif. Organs., 24, 799 (2001)
Funatsu K, Ijima H, Nakazawa K, Yamashita Y, Shimada M, Sugimachi K, Artif. Organs., 25, 194 (2001)
Yamashita Y, Shimada M, Tsujita E, Rikimaru T, Ijima H, Nakazawa K, Sakiyama R, Fukuda J, Funatsu K, Int. J. Artif. Organs., 24, 34 (2001)
Yamashita Y, Shimada M, Tsujita E, Tanaka S, Ijima H, Nakazawa K, Sakiyama R, Fukuda J, Ueda T, Funatsu K, Cell Transp., 10, 717 (2001)
Kurosawa H, Yasumoto K, Kimura T, Amano Y, Biotechnol. Bioeng., 70(2), 160 (2000)
Ijima H, Nakazawa K, Koyama S, Kaneko M, Matsushita T, Gion T, Shirabe K, Shimada M, Takenaka K, Sugimachi K, Funatsu K, Int. J. Artif. Organs., 23, 446 (2000)
Ijima H, Nakazawa K, Koyama S, Kaneko M, Matsushita T, Gion T, Shirabe K, Shimada M, Takenaka K, Sugimachi K, Funatsu K, Int. J. Artif. Organs., 23, 389 (2000)
Fujii Y, Nakazawa K, Funatsu K, J. Biomater. Sci.-Polym. Ed., 11, 731 (2000)
Ijima H, Nakazawa K, Mizumoto H, Matsushita T, Funatsu K, J. Biomater. Sci.-Polym. Ed., 9, 765 (1998)
Matsushita T, Ijima H, Koide N, Funatsu K, Appl. Microbiol. Biotechnol., 36, 324 (1991)
Ikada Y, Okano T, Tissue Engineering for Therapeutic Use 3, Elsevier Scientific Publishing Co., Amsterdam (1999)
Kaneko M, Fukuda J, Ijima H, Nakazawa K, Gion T, Shimada M, Shirabe K, Takenaka K, Sugimachi K, Funatsu K, Mater. Sci. Eng., C6, 245 (1998)
Ijima H, Matsushita T, Nakazawa K, Fujii Y, Funatsu K, Tissue. Eng., 4, 213 (1998)
Gion T, Shimada M, Shimada M, Shirabe K, Nakazawa K, Ijima H, Matsushita T, Funatsu K, Sugimachi K, J. Surg. Res., 82, 131 (1999)
Sato Y, Ochiya T, Yasuda Y, Matsubara K, Hepatology, 19, 1023 (1994)
Rowley JA, Madlabayan G, Mooney DJ, Biomaterials, 20, 45 (1999)
Lopina ST, Wu G, Merrill EW, Cima LG, Biomaterials, 17, 559 (1996)
Matsuda T, Moghaddam MJ, J. Polym. Sci. A: Polym. Chem., 31, 1589 (1993)
Lee YC, Lee RT, Neoglycoconjugates: Preparation and Application, Academic Press, San Diego (1994)
Nathan BJ, Evans PJ, Cryobiology, 40, 176 (2000)
Evans PJ, Cell. Biol. Intern., 19, 855 (1995)
Zmora S, Glicklis R, Cohen S, Biomaterials, 23, 4087 (2002)
Yang J, Goto M, Ise H, Cho CS, Akaike T, Biomaterials, 23, 471 (2002)
Ikada Y, Okano T, Tissue Engineering for Therapeutic Use 3, Elsevier Scientific Publishing Co., Amsterdam (1999)
Ohshima N, Yanagi K, Miyoshi H, Artif. Organs., 21, 1169 (1997)
Yanagi K, Mizuno S, Ohshima N, Trans. Am. Soc. Artif. Intern. Organs., 36, 727 (1990)
Yanagi K, Miyoshi H, Fukuda H, Ohshima N, Appl. Microbiol. Biotechnol., 37, 316 (1992)
Lee SJ, Lee YM, Khang G, Kim IY, Lee B, Lee HB, Macromol. Res., 10(3), 150 (2002)
Khang G, Park CS, Rhee JM, Lee SJ, Lee YM, Choi MK, Lee HB, Lee I, Korea Polym. J., 9(5), 267 (2001)
Seglen PO, Methods Cell Biol., 13, 23 (1976)
Gutsche AT, Zurlo HL, Yager JJ, Leong KW, Biomaterials, 17, 387 (1996)
Yang J, Chung TW, Nagaoka M, Goto M, Cho CS, Akaike T, Biotechnol. Lett., 23(17), 1385 (2001)

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[13] Kaneko M, Fukuda J, Ijima H, Nakazawa K, Gion T, Shimada M, Shirabe K, Takenaka K, Sugimachi K, Funatsu K, Mater. Sci. Eng., C6, 245 (1998)

[14] Ijima H, Matsushita T, Nakazawa K, Fujii Y, Funatsu K, Tissue. Eng., 4, 213 (1998)

[15] Gion T, Shimada M, Shimada M, Shirabe K, Nakazawa K, Ijima H, Matsushita T, Funatsu K, Sugimachi K, J. Surg. Res., 82, 131 (1999)

[16] Sato Y, Ochiya T, Yasuda Y, Matsubara K, Hepatology, 19, 1023 (1994)

[17] Rowley JA, Madlabayan G, Mooney DJ, Biomaterials, 20, 45 (1999)

[18] Lopina ST, Wu G, Merrill EW, Cima LG, Biomaterials, 17, 559 (1996)

[19] Matsuda T, Moghaddam MJ, J. Polym. Sci. A: Polym. Chem., 31, 1589 (1993)

[20] Lee YC, Lee RT, Neoglycoconjugates: Preparation and Application, Academic Press, San Diego (1994)

[21] Nathan BJ, Evans PJ, Cryobiology, 40, 176 (2000)

[22] Evans PJ, Cell. Biol. Intern., 19, 855 (1995)

[23] Zmora S, Glicklis R, Cohen S, Biomaterials, 23, 4087 (2002)

[24] Yang J, Goto M, Ise H, Cho CS, Akaike T, Biomaterials, 23, 471 (2002)

[25] Ikada Y, Okano T, Tissue Engineering for Therapeutic Use 3, Elsevier Scientific Publishing Co., Amsterdam (1999)

[26] Ohshima N, Yanagi K, Miyoshi H, Artif. Organs., 21, 1169 (1997)

[27] Yanagi K, Mizuno S, Ohshima N, Trans. Am. Soc. Artif. Intern. Organs., 36, 727 (1990)

[28] Yanagi K, Miyoshi H, Fukuda H, Ohshima N, Appl. Microbiol. Biotechnol., 37, 316 (1992)

[29] Lee SJ, Lee YM, Khang G, Kim IY, Lee B, Lee HB, Macromol. Res., 10(3), 150 (2002)

[30] Khang G, Park CS, Rhee JM, Lee SJ, Lee YM, Choi MK, Lee HB, Lee I, Korea Polym. J., 9(5), 267 (2001)

[31] Seglen PO, Methods Cell Biol., 13, 23 (1976)

[32] Gutsche AT, Zurlo HL, Yager JJ, Leong KW, Biomaterials, 17, 387 (1996)

[33] Yang J, Chung TW, Nagaoka M, Goto M, Cho CS, Akaike T, Biotechnol. Lett., 23(17), 1385 (2001)