Fluidity of biodegradable substrate regulates carcinoma cell behavior: A novel approach to cancer therapy

Sharmy S Mano; Koichiro Uto; Takao Aoyagi; Mitsuhiro Ebara; Sharmy S Mano; Koichiro Uto; Takao Aoyagi; Mitsuhiro Ebara

doi:10.3934/matersci.2016.1.66

AIMS Materials Science

2016, Volume 3, Issue 1: 66-82. doi: 10.3934/matersci.2016.1.66

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Fluidity of biodegradable substrate regulates carcinoma cell behavior: A novel approach to cancer therapy

1.
Smart Biomaterials Group, Biomaterials Unit, International Center for Materials Nanoarchitectonics (WPI-MANA), National Institute for Materials Science (NIMS), 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan
2.
Present Address: Department of Bioengineering, University of Washington, 3720 15th Ave NE, Seattle, WA 98195, United States

Received: 12 November 2015 Accepted: 10 January 2016 Published: 12 January 2016

Although various polymeric substrates with different stiffness have been applied for the regulation of cells’ fate, little attention has been given to the effects of substrates’ fluidity. Here, we implement for the first time biodegradable polymer with fluidic property for cancer therapy by investigating cell adhesion, proliferation, apoptosis/death, cycles of cancer cells as well as the anticancer drug efficacy. To achieve this, we prepared crosslinked and non-crosslinked copolymers of ɛ-caprolactone-co-D, L-lactide (P(CL-co-DLLA)). The tensile test showed the crosslinked P(CL-co-DLLA) substrate has the stiffness of 261 kPa while the loss modulus G’’ of the non-crosslinked substrate is always higher than the storage modulus G’ (G’’/G’=3.06), indicating a quasi-liquid state. Human lung epithelial adenocarcinoma cells on crosslinked substrate showed well- spread actin stress fibers and visible focal adhesion with an increased S phase (decreased G0/G1 phase). The cells on non-crosslinked substrate, on the other hand, showed rounded morphology without visible focal adhesion and an accumulated G0/G1 phase (decreased S phase). These results suggest that the behavior of cancer cells not only depends on stiffness but also the fluidity of P(CL-co-DLLA) substrate. In addition, the effects of substrate’s fluidity on anti-cancer drug efficacy were also investigated. The IC₅₀ values of paclitaxel for cancer cells on crosslinked and non-crosslinked substrates are 5.46 and 2.86 nM, respectively. These results clearly indicate that the fluidity of polymeric materials should be considered as one of the crucial factors to study cellular functions and molecular mechanism of cancer progression.
- fluidity,
- poly (ɛ-caprolactone),
- cancer cells,
- cell cycle,
- paclitaxel
Citation: Sharmy S Mano, Koichiro Uto, Takao Aoyagi, Mitsuhiro Ebara. Fluidity of biodegradable substrate regulates carcinoma cell behavior: A novel approach to cancer therapy[J]. AIMS Materials Science, 2016, 3(1): 66-82. doi: 10.3934/matersci.2016.1.66

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Abstract

Although various polymeric substrates with different stiffness have been applied for the regulation of cells’ fate, little attention has been given to the effects of substrates’ fluidity. Here, we implement for the first time biodegradable polymer with fluidic property for cancer therapy by investigating cell adhesion, proliferation, apoptosis/death, cycles of cancer cells as well as the anticancer drug efficacy. To achieve this, we prepared crosslinked and non-crosslinked copolymers of ɛ-caprolactone-co-D, L-lactide (P(CL-co-DLLA)). The tensile test showed the crosslinked P(CL-co-DLLA) substrate has the stiffness of 261 kPa while the loss modulus G’’ of the non-crosslinked substrate is always higher than the storage modulus G’ (G’’/G’=3.06), indicating a quasi-liquid state. Human lung epithelial adenocarcinoma cells on crosslinked substrate showed well- spread actin stress fibers and visible focal adhesion with an increased S phase (decreased G0/G1 phase). The cells on non-crosslinked substrate, on the other hand, showed rounded morphology without visible focal adhesion and an accumulated G0/G1 phase (decreased S phase). These results suggest that the behavior of cancer cells not only depends on stiffness but also the fluidity of P(CL-co-DLLA) substrate. In addition, the effects of substrate’s fluidity on anti-cancer drug efficacy were also investigated. The IC₅₀ values of paclitaxel for cancer cells on crosslinked and non-crosslinked substrates are 5.46 and 2.86 nM, respectively. These results clearly indicate that the fluidity of polymeric materials should be considered as one of the crucial factors to study cellular functions and molecular mechanism of cancer progression.

References

[1]	Nishimura N, Sasaki T (2008) Regulation of epithelial cell adhesion and repulsion: role of endocytic recycling. J Med Invest 55: 9–15. doi: 10.2152/jmi.55.9
[2]	Sheetz MP, Felsenfeld DP, Galbraith CG (1998) Cell migration: regulation of force on extracellular-matrix-integrin complexes. Trends Cell Biol 8: 51–54.
[3]	Koohestani F, Braundmeier AG, Mahdian A, et al. (2013) Extracellular matrix collagen alters cell proliferation and cell cycle progression of human uterine leiomyoma smooth muscle cells. PLOS ONE 8: e75844. doi: 10.1371/journal.pone.0075844
[4]	Philp D, Chen SS, Fitzgerald W, et al. (2005) Complex extracellular matrices promote tissue-specific stem cell differentiation. Stem Cells 23: 288–296. doi: 10.1634/stemcells.2002-0109
[5]	Farrelly N, Lee YJ, Oliver J, et al. (1999) Extracellular matrix regulates apoptosis in mammary epithelium through a control on insulin signaling. J Cell Biol 144: 1337–1347. doi: 10.1083/jcb.144.6.1337
[6]	Lu P, Weaver VM, Werb Z (2012) The extracellular matrix: a dynamic niche in cancer progression. J Cell Biol 196: 395–406. doi: 10.1083/jcb.201102147
[7]	Pickup MW, Mouw JK, Weaver VM (2014) The extracellular matrix modulates the hallmarks of cancer. EMBO Reports 15: 1243–1253. doi: 10.15252/embr.201439246
[8]	Ulrich TA, Pardo EMDJ, Kumar S (2009) The mechanical rigidity of the extracellular matrix regulates the structure, motility, and proliferation of glioma cells. Cancer Res 69: 4167–74.
[9]	Yeung T, Georges PC, Flanagan LA, et al. (2005) Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil Cytoskel 60: 24–34. doi: 10.1002/cm.20041
[10]	Tilghman WR, Blais EM, Cowan CR, et al. (2012) Matrix rigidity regulates cancer cell growth by modulating cellular metabolism and protein synthesis. PLOS ONE 7: e37231. doi: 10.1371/journal.pone.0037231
[11]	Pathak A, Kumar S (2012) Independent regulation of tumor cell migration by matrix stiffness and confinement. Proc Natl Acad Sci U S A 109: 10334–10339 doi: 10.1073/pnas.1118073109
[12]	Liu J, Tan Y, Zhang H, et al. (2012) Soft fibrin gels promote selection and growth of tumorigenic cells. Nat Mat 11: 734–741. doi: 10.1038/nmat3361
[13]	Ghosh K, Ingber DE (2007) Micromechanical control of cell and tissue development: Implications for tissue engineering. Advan Drug Deliv Rev 59: 1306–1318. doi: 10.1016/j.addr.2007.08.014
[14]	Eshraghi S, Das S (2012) Micromechanical finite element modeling and experimental characterization of the compressive mechanical properties of polycaprolactone: hydroxyapatite composite scaffolds prepared by selective laser sintering for bone tissue engineering. Acta Biomater 8: 3138–3143. doi: 10.1016/j.actbio.2012.04.022
[15]	Duncan R (2003) The dawning era of polymer therapeutics. Nat Rev Drug Discov 2: 347−360.
[16]	Peer D, Karp JM, Hong S, et al. (2007) Nanocarriers as an emerging platform for cancer therapy. Nat Nanotech 2: 751–760. doi: 10.1038/nnano.2007.387
[17]	Krishnan V, Xu X, Barwe SP, et al. (2013) Dexamethasone-loaded block copolymer nanoparticles induce leukemia cell death and enhance therapeutic efficacy: A novel application in pediatric nanomedicine. Mol Pharmaceutics 10: 2199−2210.
[18]	Pathak A, Kumar S (2013) Dual anticancer drug/superparamagnetic iron oxide-loaded PLGA-based nanoparticles for cancer therapy and magnetic resonance imaging. Int J Pharm 447: 94–101. doi: 10.1016/j.ijpharm.2013.02.042
[19]	Woodruff MA, Hutmacher DW (2010) The return of a forgotten polymer: Polycaprolactone in the 21st century. Progress in Polymer Sci 35: 1217–1256. doi: 10.1016/j.progpolymsci.2010.04.002
[20]	Abedalwafa M, Wang F, Wang L, et al. (2013) Biodegradable poly-epsilon-caprolactone (PCL) for tissue engineering applications: a review. Rev Adv Mater Sci 34: 123–140.
[21]	Rie JV, Declercq H, Hoorick JV, et al. (2015) Cryogel-PCL combination scaffolds for bone tissue repair. J Mater Sci Mater Med 26:123. doi: 10.1007/s10856-015-5465-8
[22]	Uto K, Muroya T, Okamoto M, et al. (2012) Design of super-elastic biodegradable scaffolds with longitudinally oriented microchannels and optimization of the channel size for schwann cell migration. Sci Technol Adv Mater 13: 064207. doi: 10.1088/1468-6996/13/6/064207
[23]	Uto K, Yamamoto K, Hirase S, et al. (2006) Temperature-responsive cross-linked poly(ε-caprolactone) membrane that functions near body temperature. J Control Release 110: 408–413. doi: 10.1016/j.jconrel.2005.10.024
[24]	Ebara M, Uto K, Idota N, et al. (2012) Shape-memory surface with dynamically tunable nano-geometry activated by body heat. Adv Mater 24: 273–278. doi: 10.1002/adma.201102181
[25]	Versaevel M, Grevesse T, Gabriele S (2012) Spatial coordination between cell and nuclear shape within micropatterned endothelial cells. Nat Commun 3: 671. doi: 10.1038/ncomms1668
[26]	Forte G, Pagliari S, Ebara M, et al. (2012) Substrate stiffness modulates gene expression and phenotype in neonatal cardiomyocytes in vitro. Tissue Eng Part A 18: 1837–1848. doi: 10.1089/ten.tea.2011.0707
[27]	Romanazzo S, Forte G, Ebara M, et al. (2012) Substrate stiffness affects skeletal myoblast differentiation in vitro. Sci Technol Adv Mater 13: 064211. doi: 10.1088/1468-6996/13/6/064211
[28]	Uto K, Ebara M, Aoyagi T (2014) Temperature-responsive poly(ε-caprolactone) cell culture platform with dynamically tunable nano-roughness and elasticity for control of myoblast morphology. Int J Mol Sci 15: 1511–1524. doi: 10.3390/ijms15011511
[29]	Sell SA, Wolfe PS, Garg K (2010) The use of natural polymers in tissue engineering: a focus on electrospun extracellular matrix analogues. Polymers 2: 522–553. doi: 10.3390/polym2040522
[30]	Gunatillake PA, Adhikari R (2003) Biodegradable synthetic polymers for tissue engineering. European Cells and Mat 5: 1–16.
[31]	Breuls RGM, Jiya TU, Smit TH (2008) Scaffold stiffness influences cell behavior: opportunities for skeletal tissue engineering. Open Orthopedics 2: 103–109. doi: 10.2174/1874325000802010103
[32]	Park JS, Chu JS, Tsou AD (2011) The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-b. Biomaterials 32: 3921–3930.
[33]	Ni Y, Chiang MYM (2007) Cell morphology and migration linked to substrate rigidity. Soft Matter 3: 1285–1292.
[34]	Yeung T, Georges PC, Flanagan LA (2005) Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil Cytoskeleton 60: 24–34.
[35]	Tilghman RW, Cowan CR, Mih JD, et al. (2010) Matrix rigidity regulates cancer cell growth and cellular phenotype. PLOS ONE 5: 9.
[36]	Wozniak MA, Modzelewska K, Kwong L, et al. (2004) Focal adhesion regulation of cell behavior. Biochim Biophys Acta 1692: 103–119. doi: 10.1016/j.bbamcr.2004.04.007
[37]	Kroemer G, Galluzzi L, Vandenabeele P, et al. (2009) Classification of cell death: recommendations of the nomenclature committee on cell death. Cell Death Differ 16: 3–11. doi: 10.1038/cdd.2008.150
[38]	Campisi J, Fagagna FDD (2007) Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 8: 729–740. doi: 10.1038/nrm2233
[39]	Chen QM, Liu J, Merrett JB (2000) Apoptosis or senescence-like growth arrest: influence of cell-cycle position, p53, p21 and bax in H₂O₂ response of normal human fibroblasts. Biochem J 15: 543–551.
[40]	Assoian RK, Klein EA (2008) Growth control by intracellular tension and extracellular stiffness. Trends Cell Biol 18: 347–352. doi: 10.1016/j.tcb.2008.05.002
[41]	Vicencio JM, Galluzzi L, Tajeddine N, et al. (2008) Senescence, apoptosis or autophagy? when a damaged cell must decide its path- A mini review. Gerontology 54: 92–99.
[42]	Johnson DG, Walker CL (1999) Cyclins and cell cycle checkpoints. Annu Rev Pharm Tox 39: 295–312. doi: 10.1146/annurev.pharmtox.39.1.295
[43]	Davis PK, Ho A, Dowdy SF (2001) Biological methods for cell-cycle synchronization of mammalian cells. Bio Techniques 30: 1322–1331.
[44]	Tian Y, Luo C, Lu Y, et al. (2012) Cell cycle synchronization by nutrient modulation, Integr Biol (Camb) 4: 328–334.
[45]	Lee WC, Bhagat AAS, Huang S, et al. (2011) High-throughput cell cycle synchronization using inertial forces in spiral microchannels. Lab Chip 11: 1359–1367. doi: 10.1039/c0lc00579g
[46]	Chen M, Huang J, Yang X, et al. (2012) Serum starvation induced cell cycle synchronization facilitates human somatic cells reprogramming. PLOS ONE 7: e28203. doi: 10.1371/journal.pone.0028203
[47]	Gstraunthaler G (2003) Alternatives to the use of fetal bovine serum: Serum-free cell culture, ALTEX 20: 275–281.
[48]	Eric AK, Liqun Y, Devashish K, et al. (2009) Cell-cycle control by physiological matrix elasticity and in vivo tissue stiffening. Curr Biol 19: 1511–1518. doi: 10.1016/j.cub.2009.07.069
[49]	Özdemir O (2011) Negative impact of paclitaxel crystallization on hydrogels and novel approaches for anticancer drug delivery systems, Current cancer treatment- Novel beyond conventional approaches. In Tech Open, Croatia 767–782
[50]	Chiang PC, Goul S, Nannini M (2014) Nanosuspension delivery of paclitaxel to xenograft mice can alter drug disposition and anti-tumor activity. Nanoscale Res Lett 9: 156. doi: 10.1186/1556-276X-9-156
[51]	Liebmann JE, Cook JA, Lipschultz C, et al. (1993) Cytotoxic studies of pacfitaxel (Taxol®) in human tumour cell lines. Br J Cancer 68: 1104–1109. doi: 10.1038/bjc.1993.488

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