In vitro micro-mineralized tissue formation by the combinatory condition of adipose-derived stem cells, macroporous PLGA microspheres and a bioreactor

Min-Seo Jung; Han Byul Jang; Soo-Eon Lee; Jae-Hong Park; Yu-Shik Hwang

Macromolecular Research, Vol.22, No.1, 47-57, January, 2014

DOI10.1007/s13233-014-2002-4 Export Citation

In vitro micro-mineralized tissue formation by the combinatory condition of adipose-derived stem cells, macroporous PLGA microspheres and a bioreactor

Min-Seo Jung, Han Byul Jang¹, Soo-Eon Lee¹, Jae-Hong Park¹, and Yu-Shik Hwang^†

Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, Kyung Hee University, Seoul, 130-701, Korea
¹Department of Pediatric Dentistry, School of Dentistry, Kyung Hee University, Seoul, 130-701, Korea

E-mail:

For the final purpose of accelerating the restoration of defected bone tissue, researches for developing in vitro three dimensional (3D) mineralized tissue using various stem cells, scaffolds and culture systems have been extensively done. In this research, an integrated bioprocess to generate stem cell-based 3D mineralized construct was developed using adipose-derived stem cell (ADSC), highly porous biodegradable poly(D,L-lactide-co-glycolide) (PLGA) microspheres and a high-aspect ratio vessel (HARV) bioreactor system. First, ADSCs adhered uniformly on poly-L-ornithine-coated macroporous PLGA microspheres, and the ADSC/microsphere composites were cultured in the presence of osteogenic supplements in a HARV bioreactor. Alkaline phosphatase (ALPase) activity assay, immunocytochemical staining and quantitative real-time polymerase chain reaction (qPCR) analysis showed temporal increase of ALPase activity, molecular expressions of type I collagen, osteocalcin and runx2 and upregulation of runx2, Sp7, type I collagen and bone sialoprotein mRNA during 3D osteogenic culture of ADSC/microsphere composites. Finally, 3D dynamic osteogenic culture generated highly mineralized micro tissues as validated by alizarin red-S staining and SEM-EDS. The results demonstrated that cell-based 3D micro-mineralized tissue could be generated by integrated bioprocess, and potentially utilized for bone tissue regeneration. The integrated bioprocess in this study may provide an efficient and scalable culture system for application in bone tissue engineering.

Keywords:adipose-derived stem cell (ADSC);microsphere;bioreactor;osteogenic;mineralization;micro-tissue

[References]

Ceuninck FD, Lesur C, Pastoureau P, Caliez A, Sabatini M, Methods Mol. Med., 100, 15 (2004)
Hwang NS, Kim MS, Sampattavanich S, Baek JH, Zhang Z, Elisseeff J, Stem Cells, 24, 284 (2006)
Ishaug-Riley SL, Crane-Kruger GM, Yaszemski MJ, Mikos AG, Biomaterials, 19, 1405 (1998)
Holy CE, Shoichet MS, Davies JE, J. Biomed. Mater. Res., 51, 376 (2000)
Glowacki J, Mizuno S, Greenberger JS, Cell Transplant., 7, 319 (1998)
Mueller SM, Mizuno S, Gerstenfeld LC, Glowacki J, J. Bone Miner. Res., 14, 2118 (1999)
Qiu QQ, Ducheyne P, Ayyaswamy PS, Biomaterials, 20, 989 (1999)
Botchwey EA, Pollack SR, Levine EM, Laurencin CT, J. Biomed. Mater. Res., 55, 242 (2001)
Hwang YS, Cho J, Chan KLA, Heng J, Boccacini AR, Kazarian SG, Williams D, Polak JM, Mantalaris A, Biomaterials, 30, 499 (2009)
Hammond TG, Hammond JM, Am. J. Physiol. Renal. Physiol., 281, 12 (2001)
Pollack SR, Meany DF, Levine EM, Litt M, Johnston ED, Tissue Eng., 6, 519 (2000)
Wang Y, Umera T, Dong J, Kojima H, Tanaka J, Tateichi T, Tissue Eng., 9, 1205 (2003)
Kim TK, Yoon JJ, Lee DS, Park TG, Biomaterials, 27, 152 (2006)
Betz RR, Orthopaedics, 25, S561 (2002)
Petite H, Viateau V, Bensaid W, Meunier A, de Pollak C, Bourguignon M, Oudina K, Sedel L, Guillemin G, Nat. Biotechnol., 18, 959 (2000)
Tamura S, Kataoka H, Matsui Y, Shionoya Y, Ohno K, Michi KI, Bone, 29, 169 (2001)
Karen JLB, Scott P, James FK, Biomaterials, 21, 2347 (2000)
Hutmacher DW, Biomaterials, 21, 2529 (2000)
Blaker JJ, Maquet V, Jerome R, Boccaccini AR, Nazhat SN, Acta Biomater., 1, 643 (2005)
Zhang ZY, Teoh SH, Teo EY, Chong MSK, Shin CW, Tien FT, Choolani MA, Chan JKY, Biomaterials, 31, 8684 (2010)
Chung HH, Kim HK, Yoon JJ, Park TG, Pharm. Res., 23, 1835 (2006)
Huang CC, Wei HJ, Yeh YC, Wang JJ, Lin WW, Lee TY, Hwang SM, Choi SW, Xia Y, Chang Y, Sung HW, Biomaterials, 33, 4069 (2012)
Kang SW, Seo SW, Choi CY, Kim BS, Tissue Eng. Part C, 14, 25 (2008)
Kang SW, La WG, Kim BS, J. Biomater. Sci.-Polym. Ed., 20, 399 (2009)
Harper MM, Ye EA, Blong CC, Jacobson ML, Sakaguchi DS, J. Mol. Neurosci., 40, 269 (2010)
Song JM, Kim BC, Park JH, Kwon IK, Mantalaris A, Hwang YS, J. Biomed. Mater. Res., 5, 1 (2010)
Seeman E, Delmas PD, N. Engl. J. Med, 354, 2250 (2006)
Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G, Cell, 89, 747 (1997)
Banerjee C, Javed A, Choi JY, Green J, Rosen V, van Wijnen AJ, Stein JL, Lian JE, Stein GS, Endocrinology, 142, 4026 (2001)
Viereck V, Siqqelkow H, Tauber S, Raddatz D, Schutze N, Hufner M, J. Cell. Biochem., 86, 348 (2002)
Tracy MA, Ward KL, Firouzabadian L, Wang Y, Dong N, Qian R, Zhang Y, Biomaterials, 20, 1057 (1999)
Sugawara Y, Suzuki K, Koshikawa M, Ando M, Iida J, Jpn. J. Pharmacol., 88, 262 (2002)
Yoshikawa M, Suzuki K, Kajii T, Koshikawa M, Imai T, Matsumoto A, J. Hard Tissue Biol., 8, 37 (1999)

[1] Ceuninck FD, Lesur C, Pastoureau P, Caliez A, Sabatini M, Methods Mol. Med., 100, 15 (2004)

[2] Hwang NS, Kim MS, Sampattavanich S, Baek JH, Zhang Z, Elisseeff J, Stem Cells, 24, 284 (2006)

[3] Ishaug-Riley SL, Crane-Kruger GM, Yaszemski MJ, Mikos AG, Biomaterials, 19, 1405 (1998)

[4] Holy CE, Shoichet MS, Davies JE, J. Biomed. Mater. Res., 51, 376 (2000)

[5] Glowacki J, Mizuno S, Greenberger JS, Cell Transplant., 7, 319 (1998)

[6] Mueller SM, Mizuno S, Gerstenfeld LC, Glowacki J, J. Bone Miner. Res., 14, 2118 (1999)

[7] Qiu QQ, Ducheyne P, Ayyaswamy PS, Biomaterials, 20, 989 (1999)

[8] Botchwey EA, Pollack SR, Levine EM, Laurencin CT, J. Biomed. Mater. Res., 55, 242 (2001)

[9] Hwang YS, Cho J, Chan KLA, Heng J, Boccacini AR, Kazarian SG, Williams D, Polak JM, Mantalaris A, Biomaterials, 30, 499 (2009)

[10] Hammond TG, Hammond JM, Am. J. Physiol. Renal. Physiol., 281, 12 (2001)

[11] Pollack SR, Meany DF, Levine EM, Litt M, Johnston ED, Tissue Eng., 6, 519 (2000)

[12] Wang Y, Umera T, Dong J, Kojima H, Tanaka J, Tateichi T, Tissue Eng., 9, 1205 (2003)

[13] Kim TK, Yoon JJ, Lee DS, Park TG, Biomaterials, 27, 152 (2006)

[14] Betz RR, Orthopaedics, 25, S561 (2002)

[15] Petite H, Viateau V, Bensaid W, Meunier A, de Pollak C, Bourguignon M, Oudina K, Sedel L, Guillemin G, Nat. Biotechnol., 18, 959 (2000)

[16] Tamura S, Kataoka H, Matsui Y, Shionoya Y, Ohno K, Michi KI, Bone, 29, 169 (2001)

[17] Karen JLB, Scott P, James FK, Biomaterials, 21, 2347 (2000)

[18] Hutmacher DW, Biomaterials, 21, 2529 (2000)

[19] Blaker JJ, Maquet V, Jerome R, Boccaccini AR, Nazhat SN, Acta Biomater., 1, 643 (2005)

[20] Zhang ZY, Teoh SH, Teo EY, Chong MSK, Shin CW, Tien FT, Choolani MA, Chan JKY, Biomaterials, 31, 8684 (2010)

[21] Chung HH, Kim HK, Yoon JJ, Park TG, Pharm. Res., 23, 1835 (2006)

[22] Huang CC, Wei HJ, Yeh YC, Wang JJ, Lin WW, Lee TY, Hwang SM, Choi SW, Xia Y, Chang Y, Sung HW, Biomaterials, 33, 4069 (2012)

[23] Kang SW, Seo SW, Choi CY, Kim BS, Tissue Eng. Part C, 14, 25 (2008)

[24] Kang SW, La WG, Kim BS, J. Biomater. Sci.-Polym. Ed., 20, 399 (2009)

[25] Harper MM, Ye EA, Blong CC, Jacobson ML, Sakaguchi DS, J. Mol. Neurosci., 40, 269 (2010)

[26] Song JM, Kim BC, Park JH, Kwon IK, Mantalaris A, Hwang YS, J. Biomed. Mater. Res., 5, 1 (2010)

[27] Seeman E, Delmas PD, N. Engl. J. Med, 354, 2250 (2006)

[28] Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G, Cell, 89, 747 (1997)

[29] Banerjee C, Javed A, Choi JY, Green J, Rosen V, van Wijnen AJ, Stein JL, Lian JE, Stein GS, Endocrinology, 142, 4026 (2001)

[30] Viereck V, Siqqelkow H, Tauber S, Raddatz D, Schutze N, Hufner M, J. Cell. Biochem., 86, 348 (2002)

[31] Tracy MA, Ward KL, Firouzabadian L, Wang Y, Dong N, Qian R, Zhang Y, Biomaterials, 20, 1057 (1999)

[32] Sugawara Y, Suzuki K, Koshikawa M, Ando M, Iida J, Jpn. J. Pharmacol., 88, 262 (2002)

[33] Yoshikawa M, Suzuki K, Kajii T, Koshikawa M, Imai T, Matsumoto A, J. Hard Tissue Biol., 8, 37 (1999)