Preparation of novel RGD-conjugated thermosensitive mPEG-PCL composite hydrogels and in vitro investigation of their impacts on adhesion-dependent cellular behavior

Hyun Joo Kim; Su Jung You; Dae Hyeok Yang; Heung Jae Chun; Moon Suk Kim

Journal of Industrial and Engineering Chemistry, Vol.84, 226-235, April, 2020

DOI10.1016/j.jiec.2020.01.001 Export Citation

Preparation of novel RGD-conjugated thermosensitive mPEG-PCL composite hydrogels and in vitro investigation of their impacts on adhesion-dependent cellular behavior

Hyun Joo Kim^{1, 2}, Su Jung You², Dae Hyeok Yang², Heung Jae Chun^{1, 2, †}, and Moon Suk Kim³

¹Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea
²Institute of Cell and Tissue Engineering, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea
³Department of Molecular Science and Technology, Ajou University, Suwon, 16499, Republic of Korea

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The arginine?glycine?aspartic acid (RGD) motif plays a critical role in cell-extracellular matrix (ECM) interactions through focal adhesion, which may orchestrate various aspects of cellular behaviors for development and maintenance of tissues. Here, we prepared novel thermosensitive hydrogel systems comprising methoxy polyethylene glycol-b-polycaprolactone (MP) and RGD-conjugated MP (MP-RGD) with various RGD contents (MP/MP-RGD) based on our recently patented technology. The conjugation of RGD to MP through biomolecular nucleophilic substitution was qualitatively and quantitatively assessed using 1H nuclear magnetic resonance (NMR) spectroscopy and the Sakaguchi assay, respectively. The sol?gel?sol transition with increasing temperature was confirmed using viscoelastic measurements together with tilting tests. In vitro experiments using bone marrow-derived mesenchymal stem cells (BMSCs) revealed that MP/MP-RGD hydrogels significantly enhanced expression of integrins (α2, α5, and β1) and facilitated formation of focal adhesions (increased expression of FAK, focal adhesion kinase) that lead to cytoskeletal reorganization through integrin-mediated cell signaling. Real-time polymerase chain reaction (PCR) array also showed upregulation of signaling genes involved in the receptor tyrosine kinase (RTK) signaling cascade such as mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) for growth factors and phosphoinositide 3-kinase (PI3K) and AKT (protein kinase B) for survival factors. This suggests that extensive crosstalk pathways exist between integrins and RTKs, which was confirmed through western blotting. Consequently, MP/MP-RGD hydrogels showed a significantly increased BMSC proliferation rate with increasing RGD content of the samples.

Keywords:BMSCs;RGD;Integrin;Focal adhesion;In situ-forming hydrogel;Thermosensitive hydrogel

[References]

Liang K, Bae KH, Kurisawa M, J. Mater. Chem. B, 7, 3775 (2019)
Kim MS, Kim JH, Min BH, Chun HJ, Han DK, Lee HB, Polym. Rev., 51, 23 (2011)
Kang YM, Lee SH, Lee JY, Son JS, Kim BS, Lee B, Chun HJ, Min BH, Kim JH, Kim MS, Biomaterials, 31, 2453 (2010)
Hyun H, Kim YH, Song IB, Lee JW, Kim MS, Khang G, Park K, Lee HB, Biomacromolecules, 8(4), 1093 (2007)
Ahn HH, Kim KS, Lee JH, Lee JY, Kim BS, Lee IW, Chun HJ, Kim JH, Lee HB, Kim MS, Tissue Eng. Part A, 15, 1821 (2009)
Kwon JS, Yoon SM, Kwon DY, Kim DY, Tai GZ, Jin LM, Song B, Lee B, Kim JH, Han DK, Min BH, Kim MS, J. Mater. Chem. B, 1, 3314 (2013)
Kim MS, Seo KS, Khang G, Lee HB, Macromol. Rapid Commun., 26(8), 643 (2005)
Kim MS, Hyun H, Seo KS, Cho YH, Lee JW, Lee CR, Khang G, Lee HB, J. Polym. Sci. A: Polym. Chem., 44(18), 5413 (2006)
Kim MS, Kim JH, Min BH, Chun HJ, Han DK, Lee HB, Polym. Rev., 51, 23 (2011)
Huebsch N, Acta Biomater., 94, 97 (2019)
Bellis SL, Biomaterials, 32, 4205 (2011)
Lam J, Segura T, Biomaterials, 34, 3938 (2013)
Kim HJ, You SJ, Yang DH, Chun HJ, US Pat. 10226549B2 (2019). Thermosensitive biodegradable hydrogel.
Zhang Z, Lai Y, Yu L, Ding J, Biomaterials, 31, 7873 (2010)
Weber CJ, J. Biol. Chem., 86, 217 (1930)
Hyun H, Yoo YB, Kim SY, Ko HS, Chun HJ, Yang DH, J. Ind. Eng. Chem., 81, 178 (2020)
Teotia AK, Sami H, Kumar A, Structure and Design of Smart Materials, Woodhead Publishing, Oxford, pp.3 2015.
Turner CE, Nat. Cell Biol., 2, E231 (200)
Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY, Trends Biochem. Sci., 20, 448 (1995)
Milone MC, O’Doherty U, Leukemia, 32, 1529 (2018)
White M, Whittaker R, Gandara C, Stoll EA, Human Gene Therapy Methods, 28, 163 (2017)
Dittmer DP, Vahrson W, Staudt MR, Papin JR, Hines-Boykin R, Fakhari FD, Cancer Genom. Proteom., 1, 117 (2004)
Gaj S, Eijssen L, Mensink RP, Evelo CT, Genes Nutr., 3, 153 (2008)
Docheva D, Popov C, Mutschler W, Schieker M, J. Cell. Mol. Med., 11, 21 (2007)
Lv H, Li L, Sun M, Zhang Y, Chen L, Rong Y, Li Y, Stem Cell Res. Ther., 6, 103 (2015)
Chen J, Crawford R, Chen C, Xiao Y, Tissue Eng. Part B, 19, 516 (2013)
Chen S, Guo Y, Liu R, Wu S, Fang J, Huang B, Li Z, Chen Z, Chen Z, Colloids Surf. B: Biointerfaces, 164, 58 (2018)
Lee JW, Juliano R, Mol. Cells, 17, 188 (2004)

[Referenced By]

[1] Liang K, Bae KH, Kurisawa M, J. Mater. Chem. B, 7, 3775 (2019)

[2] Kim MS, Kim JH, Min BH, Chun HJ, Han DK, Lee HB, Polym. Rev., 51, 23 (2011)

[3] Kang YM, Lee SH, Lee JY, Son JS, Kim BS, Lee B, Chun HJ, Min BH, Kim JH, Kim MS, Biomaterials, 31, 2453 (2010)

[4] Hyun H, Kim YH, Song IB, Lee JW, Kim MS, Khang G, Park K, Lee HB, Biomacromolecules, 8(4), 1093 (2007)

[5] Ahn HH, Kim KS, Lee JH, Lee JY, Kim BS, Lee IW, Chun HJ, Kim JH, Lee HB, Kim MS, Tissue Eng. Part A, 15, 1821 (2009)

[6] Kwon JS, Yoon SM, Kwon DY, Kim DY, Tai GZ, Jin LM, Song B, Lee B, Kim JH, Han DK, Min BH, Kim MS, J. Mater. Chem. B, 1, 3314 (2013)

[7] Kim MS, Seo KS, Khang G, Lee HB, Macromol. Rapid Commun., 26(8), 643 (2005)

[8] Kim MS, Hyun H, Seo KS, Cho YH, Lee JW, Lee CR, Khang G, Lee HB, J. Polym. Sci. A: Polym. Chem., 44(18), 5413 (2006)

[9] Kim MS, Kim JH, Min BH, Chun HJ, Han DK, Lee HB, Polym. Rev., 51, 23 (2011)

[10] Huebsch N, Acta Biomater., 94, 97 (2019)

[11] Bellis SL, Biomaterials, 32, 4205 (2011)

[12] Lam J, Segura T, Biomaterials, 34, 3938 (2013)

[13] Kim HJ, You SJ, Yang DH, Chun HJ, US Pat. 10226549B2 (2019). Thermosensitive biodegradable hydrogel.

[14] Zhang Z, Lai Y, Yu L, Ding J, Biomaterials, 31, 7873 (2010)

[15] Weber CJ, J. Biol. Chem., 86, 217 (1930)

[16] Hyun H, Yoo YB, Kim SY, Ko HS, Chun HJ, Yang DH, J. Ind. Eng. Chem., 81, 178 (2020)

[17] Teotia AK, Sami H, Kumar A, Structure and Design of Smart Materials, Woodhead Publishing, Oxford, pp.3 2015.

[18] Turner CE, Nat. Cell Biol., 2, E231 (200)

[19] Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY, Trends Biochem. Sci., 20, 448 (1995)

[20] Milone MC, O’Doherty U, Leukemia, 32, 1529 (2018)

[21] White M, Whittaker R, Gandara C, Stoll EA, Human Gene Therapy Methods, 28, 163 (2017)

[22] Dittmer DP, Vahrson W, Staudt MR, Papin JR, Hines-Boykin R, Fakhari FD, Cancer Genom. Proteom., 1, 117 (2004)

[23] Gaj S, Eijssen L, Mensink RP, Evelo CT, Genes Nutr., 3, 153 (2008)

[24] Docheva D, Popov C, Mutschler W, Schieker M, J. Cell. Mol. Med., 11, 21 (2007)

[25] Lv H, Li L, Sun M, Zhang Y, Chen L, Rong Y, Li Y, Stem Cell Res. Ther., 6, 103 (2015)

[26] Chen J, Crawford R, Chen C, Xiao Y, Tissue Eng. Part B, 19, 516 (2013)

[27] Chen S, Guo Y, Liu R, Wu S, Fang J, Huang B, Li Z, Chen Z, Chen Z, Colloids Surf. B: Biointerfaces, 164, 58 (2018)

[28] Lee JW, Juliano R, Mol. Cells, 17, 188 (2004)