Macromolecular Research, Vol.23, No.11, 986-993, November, 2015
The role of demineralized bone particle in a PLGA scaffold designed to create a media equivalent for a tissue engineered blood vessel
E-mail:
Poly(lactic-co-glycolic acid) (PLGA) is a copolymer used for scaffolds in tissue engineering due to its biodegradable and biocompatible properties. Demineralized bone particle (DBP) is a natural material containing bone morphogenic proteins (BMPs). The objective of this study was to determine if DBP induced differentiation of mesenchymal stem cells (MSCs) into a smooth muscle cells (SMCs) phenotype and act in a similar manner when treated with TGF-β1 as SMCs under the same conditions. PLGA scaffolds were fabricated with or without DBP using the solvent casting/salt leaching technique. Proliferation of MSCs and SMCs was measured using the MTT assay, while the expression of the smooth muscle-specific markers α-smooth muscle actin (α-SMA), smooth muscle 22 alpha (SM22α), and calponin was measured using RT-PCR and western blotting. Results showed PLGA with DBP scaffolds increased cell proliferation of SMCs (1 and 3 days) and MSCs (3 and 7 days) compared to the PLGA only scaffolds. Smooth muscle-specific genes were significantly increased in SMCs cultured for 7 days on PLGA with DBP scaffolds compared to PLGA only scaffolds and were also significantly increased when used SMC medium containing TGF-β1. Interestingly, the smooth muscle-specific genes were not altered in MSCs by the addition of TGF-β1 to the PLGA with DBP. However, in the 6-well and PLGA only groups, TGF-β1 addition caused increased levels of SM22α and calponin mRNA. Western blotting results were similar to the mRNA levels. These results suggest that the cell growth and gene expression of SMCs were up-regulated by DBP and TGF-β1 in PLGA scaffold; however, they were inadequate for the shift of MSCs into a SMCs phenotype.
Keywords:poly(lactic-co-glycolic acid) (PLGA);smooth muscle cells;mesenchymal stem cells;demineralized bone particles;bone morphogenetic proteins
- Chen GP, Ushida T, Tateishi T, Macromol. Biosci., 2, 60 (2002)
- Athanasiou KA, Agrawal CM, Barber FA, Burkhart SS, Arthroscopy, 14, 726 (1998)
- Curran JM, Chen R, Hunt JA, Tissue Eng. Part A, 14, 715 (2008)
- Asahina I, Sampath TK, Hauschka PV, Exp. Cell Res., 222, 38 (1996)
- Ko EK, Jeong SI, Rim NG, Lee YM, Shin H, Lee BK, Tissue Eng. Part A, 14, 2105 (2008)
- Zhou S, Yates KE, Eid K, Glowacki J, J. Bone Miner. Res., 19, S346 (2004)
- Deckers MM, van Bezooijen RL, van der Horst G, Hoogendam J, van der Bent C, Papapoulos SE, Lowik CW, Endocrinology, 143, 1545 (2002)
- Narita Y, Yamawaki A, Kagami H, Ueda M, Ueda Y, Cell Tissue Res., 333, 449 (2008)
- Gong Z, Calkins G, Cheng EC, Krause D, Niklason LE, Tissue Eng. Part A, 15, 319 (2009)
- Gong ZD, Niklason LE, Faseb. J., 22, 1635 (2008)
- Shin’oka T, Matsumura G, Hibino N, Naito Y, Watanabe M, Konuma T, Sakamoto T, Nagatsu M, Kurosawa H, J. Thorac. Cardiovasc. Surg., 129, 1330 (2005)
- Yoon SJ, Kim SH, Ha HJ, Ko YK, So JW, Kim MS, Yang YI, Khang G, Rhee JM, Lee HB, Tissue Eng. Part A, 14, 539 (2008)
- Ko YK, Kim SH, Jeong JS, Ha HJ, Yoon SJ, Rhee JM, Kim MS, Lee HB, Khang G, Polym.(Korea), 31(1), 14 (2007)
- Yang J, Shi G, Bei J, Wang S, Cao Y, Shang Q, Yang G, Wang W, J. Biomed. Mater. Res., 62, 438 (2002)
- McGlohorn JB, Holder WD, Grimes LW, Thomas CB, Burg KJ, Tissue Eng., 10, 505 (2004)
- Kim SM, Kim SH, Kim CM, Oh AY, Kim GA, Lee IW, Rhee JM, Khang G, Kim MS, Lee HB, Tissue Eng. Regen. Med., 4, 589 (2007)
- Baek MO, Kim SH, So JW, Lim JY, Roh HW, Lee NR, Ryu GH, Cho Y, Lee SJ, Khang G, Tissue Eng. Regen. Med., 5, 91 (2008)
- Kim SH, Song JE, Lee D, Khang G, J. Biomater. Sci.-Polym. Ed., 23, 2153 (2012)
- Zhang S, Fantozzi I, Tigno DD, Yi ES, Platoshyn O, Thistlethwaite PA, Kriett JM, Yung G, Rubin LJ, Yuan XJ, Am. J. Physiol. Lung Cell. Mol. Physiol., 285, L740 (2003)
- Zeiter S, Lezuo P, Ito K, Biorheology, 46, 45 (2009)
- Sobue K, Hayashi K, Nishida W, Mol. Cell. Biochem., 190, 105 (1999)
- Lagna G, Ku MM, Nguyen PH, Neuman NA, Davis BN, Hata A, J. Biol. Chem., 282, 37244 (2007)
- Wang C, Yin S, Cen L, Liu QH, Liu W, Cao YL, Cui L, Tissue Eng. Part A, 16, 1201 (2010)
- Rodriguez LV, Alfonso Z, Zhang R, Leung J, Wu B, Ignarro LJ, Proc. Natl. Acad. Sci. U.S.A., 103, 12167 (2006)
- Roelen BA, Dijke P, J. Orthop. Sci., 8, 740 (2003)
- Tuan RS, Boland G, Tuli R, Arthritis Res. Ther., 5, 32 (2003)