Green tea polyphenol tailors cell adhesivity of RGD displaying surfaces: multicomponent models monitored optically

The interaction of the anti-adhesive coating, poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) and its Arg-Gly-Asp (RGD) functionalized form, PLL-g-PEG-RGD, with the green tea polyphenol, epigallocatechin-gallate (EGCg) was in situ monitored. After, the kinetics of cellular adhesion on the EGCg exposed coatings were recorded in real-time. The employed plate-based waveguide biosensor is applicable to monitor small molecule binding and sensitive to sub-nanometer scale changes in cell membrane position and cell mass distribution; while detecting the signals of thousands of adhering cells. The combination of this remarkable sensitivity and throughput opens up new avenues in testing complicated models of cell-surface interactions. The systematic studies revealed that, despite the reported excellent antifouling properties of the coatings, EGCg strongly interacted with them, and affected their cell adhesivity in a concentration dependent manner. Moreover, the differences between the effects of the fresh and oxidized EGCg solutions were first demonstrated. Using a semiempirical quantumchemical method we showed that EGCg binds to the PEG chains of PLL-g-PEG-RGD and effectively blocks the RGD sites by hydrogen bonds. The calculations supported the experimental finding that the binding is stronger for the oxidative products. Our work lead to a new model of polyphenol action on cell adhesion ligand accessibility and matrix rigidity.

[1]  L. Miele,et al.  EGCG, a major green tea catechin suppresses breast tumor angiogenesis and growth via inhibiting the activation of HIF-1α and NFκB, and VEGF expression , 2013, Vascular cell.

[2]  R. Horváth,et al.  Intensity interrogation near cutoff resonance for label-free cellular profiling , 2016, Scientific Reports.

[3]  Lance G. Laing,et al.  Label-Free Assays on the BIND System , 2004, Journal of biomolecular screening.

[4]  Yasuo Suzuki,et al.  Binding interaction between (-)-epigallocatechin gallate causes impaired spreading of cancer cells on fibrinogen. , 2013, Biomedical research.

[5]  Ye Fang,et al.  Resonant waveguide grating biosensor for living cell sensing. , 2006, Biophysical journal.

[6]  D. Mereles,et al.  Epigallocatechin-3-gallate (EGCG) for Clinical Trials: More Pitfalls than Promises? , 2011, International journal of molecular sciences.

[7]  Xiaofeng Meng,et al.  Stability, cellular uptake, biotransformation, and efflux of tea polyphenol (-)-epigallocatechin-3-gallate in HT-29 human colon adenocarcinoma cells. , 2002, Cancer research.

[8]  Yu-Li Wang,et al.  Substrate rigidity regulates the formation and maintenance of tissues. , 2006, Biophysical journal.

[9]  Yang Tian,et al.  Mechanism of action of (-)-epigallocatechin-3-gallate: auto-oxidation-dependent activation of extracellular signal-regulated kinase 1/2 in Jurkat cells. , 2014, Chinese journal of natural medicines.

[10]  J E Prenosil,et al.  Optical method for measurement of number and shape of attached cells in real time. , 1995, Cytometry.

[11]  B. Grzybowski,et al.  Label-free in situ optical monitoring of the adsorption of oppositely charged metal nanoparticles. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[12]  T. Slater,et al.  STUDIES ON SUCCINATE-TETRAZOLIUM REDUCTASE SYSTEMS. III. POINTS OF COUPLING OF FOUR DIFFERENT TETRAZOLIUM SALTS. , 1963, Biochimica et biophysica acta.

[13]  K. Schrör,et al.  The FASEB Journal express article 10.1096/fj.03-0007fje. Published online November 20, 2003. Mechanisms , 2022 .

[14]  M. Dembo,et al.  Substrate flexibility regulates growth and apoptosis of normal but not transformed cells. , 2000, American journal of physiology. Cell physiology.

[15]  M. Stoddart,et al.  Mammalian Cell Viability , 2011, Methods in Molecular Biology.

[16]  F. Denizot,et al.  Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. , 1986, Journal of immunological methods.

[17]  Janos Vörös,et al.  Systematic study of osteoblast response to nanotopography by means of nanoparticle-density gradients. , 2007, Biomaterials.

[18]  Jeremy J. Ramsden,et al.  Dependence of cancer cell adhesion kinetics on integrin ligand surface density measured by a high-throughput label-free resonant waveguide grating biosensor , 2014, Scientific Reports.

[19]  Wen-Bin Wu,et al.  Tea polyphenols inhibit rat vascular smooth muscle cell adhesion and migration on collagen and laminin via interference with cell-ECM interaction. , 2007, Journal of biomedical science.

[20]  Tatsuro Watanabe,et al.  Mechanism-based inhibition of cancer metastasis with (-)-epigallocatechin gallate. , 2014, Biochemical and biophysical research communications.

[21]  Abiche H. Dewilde,et al.  Dynamic cell adhesion and viscoelastic signatures distinguish normal from malignant human mammary cells using quartz crystal microbalance. , 2012, Analytical biochemistry.

[22]  M. Slevin,et al.  Bisphosphonate-related osteonecrosis of jaw (BRONJ): diagnostic criteria and possible pathogenic mechanisms of an unexpected anti-angiogenic side effect , 2013, Vascular cell.

[23]  J. Chung,et al.  The effects of epigallocatechin-3-gallate on extracellular matrix metabolism. , 2005, Journal of dermatological science.

[24]  R. McGuinness,et al.  Cellular Dielectric Spectroscopy: A Powerful New Approach to Label-Free Cellular Analysis , 2004, Journal of biomolecular screening.

[25]  Xiao Xu,et al.  The xCELLigence system for real-time and label-free monitoring of cell viability. , 2011, Methods in molecular biology.

[26]  M. Isemura,et al.  Inhibitory effect of epigallocatechin gallate on adhesion of murine melanoma cells to laminin. , 2001, Cancer letters.

[27]  Lin Shi,et al.  Effect of tea polyphenols on the adhesion of highly metastatic human lung carcinoma cell lines to endothelial cells in vitro. , 2012, Asian Pacific journal of cancer prevention : APJCP.

[28]  Adsorption properties of poly(l-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) at a hydrophobic interface: influence of tribological stress, pH, salt concentration, and polymer molecular weight. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[29]  S. Katiyar,et al.  EGCG inhibits mammary cancer cell migration through inhibition of nitric oxide synthase and guanylate cyclase. , 2008, Biochemical and biophysical research communications.

[30]  F. P. Altman Tetrazolium salts and formazans. , 1976, Progress in histochemistry and cytochemistry.

[31]  Brian T Cunningham,et al.  Label-free cell-based assays using photonic crystal optical biosensors. , 2011, The Analyst.

[32]  M. Textor,et al.  Relationship between interfacial forces measured by colloid-probe atomic force microscopy and protein resistance of poly(ethylene glycol)-grafted poly(L-lysine) adlayers on niobia surfaces. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[33]  Clay W Scott,et al.  Comparing label-free biosensors for pharmacological screening with cell-based functional assays. , 2010, Assay and drug development technologies.

[34]  Michihiro Hide,et al.  Real-time analysis of ligand-induced cell surface and intracellular reactions of living mast cells using a surface plasmon resonance-based biosensor. , 2002, Analytical biochemistry.

[35]  S. vandeVondele,et al.  RGD-containing peptide GCRGYGRGDSPG reduces enhancement of osteoblast differentiation by poly(L-lysine)-graft-poly(ethylene glycol)-coated titanium surfaces. , 2004, Journal of biomedical materials research. Part A.

[36]  Robert Horvath,et al.  Incubator proof miniaturized Holomonitor to in situ monitor cancer cells exposed to green tea polyphenol and preosteoblast cells adhering on nanostructured titanate surfaces: validity of the measured parameters and their corrections , 2015, Journal of biomedical optics.

[37]  James McColl,et al.  Structural hysteresis and hierarchy in adsorbed glycoproteins. , 2008, The Journal of chemical physics.

[38]  P. Roach,et al.  A study of stability of (-)-Epigallocatechin gallate (EGCG) from green tea in a frozen product , 2011 .

[39]  W. Lukosz,et al.  Sensitivity of grating couplers as integrated-optical chemical sensors , 1989 .

[40]  R. Arakawa,et al.  Analysis of oxidized epigallocatechin gallate by liquid chromatography/mass spectrometry. , 2003, Rapid communications in mass spectrometry : RCM.

[41]  J. Stewart Optimization of parameters for semiempirical methods V: Modification of NDDO approximations and application to 70 elements , 2007, Journal of molecular modeling.

[42]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[43]  H. Tachibana Green tea polyphenol sensing , 2011, Proceedings of the Japan Academy. Series B, Physical and biological sciences.

[44]  D. Liebler,et al.  Antioxidant chemistry of green tea catechins. New oxidation products of (-)-epigallocatechin gallate and (-)-epigallocatechin from their reactions with peroxyl radicals. , 2000, Chemical research in toxicology.

[45]  M. Textor,et al.  Effects of ionic strength and surface charge on protein adsorption at PEGylated surfaces. , 2005, The journal of physical chemistry. B.

[46]  G. Meacci,et al.  Cells test substrate rigidity by local contractions on submicrometer pillars , 2012, Proceedings of the National Academy of Sciences.

[47]  T. Wecker,et al.  Substrate rigidity modulates cell matrix interactions and protein expression in human trabecular meshwork cells. , 2008, Investigative ophthalmology & visual science.

[48]  S. Gupta,et al.  Green tea polyphenol epigallocatechin-3-gallate differentially modulates nuclear factor kappaB in cancer cells versus normal cells. , 2000, Archives of biochemistry and biophysics.

[49]  H. Galla,et al.  Analysis of the composite response of shear wave resonators to the attachment of mammalian cells. , 2000, Biophysical journal.

[50]  Robert Horvath,et al.  Bulk and surface sensitivity of a resonant waveguide grating imager , 2014 .

[51]  Ye Fang,et al.  Resonant Waveguide Grating Biosensor for Microarrays , 2010 .

[52]  Polyphenol Control of Cell Spreading on Glycoprotein Substrata , 2009, Journal of biomaterials science. Polymer edition.

[53]  R. Horváth,et al.  Optical Biosensors for Cell Adhesion , 2010 .

[54]  J. Wegener,et al.  Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. , 2000, Experimental cell research.

[55]  Robert Horvath,et al.  Adhesion kinetics of human primary monocytes, dendritic cells, and macrophages: Dynamic cell adhesion measurements with a label-free optical biosensor and their comparison with end-point assays. , 2016, Biointerphases.

[56]  S. Ferrari,et al.  Author contributions , 2021 .

[57]  R. Horváth,et al.  Self-assembled, nanostructured coatings for water oxidation by alternating deposition of Cu-branched peptide electrocatalysts and polyelectrolytes† †Electronic supplementary information (ESI) available: Tables S1 and S2, X-ray photoelectron spectroscopy (XPS) on LbL-ITO with Cu-3G and PLL, Fig. S1–S , 2016, Chemical science.

[58]  Akhlesh Lakhtakia,et al.  Optical Guided-wave Chemical and Biosensors II , 2009 .

[59]  Yen-Wen Chen,et al.  Cellular dielectric spectroscopy: a label-free comprehensive platform for functional evaluation of endogenous receptors. , 2006, Assay and drug development technologies.

[60]  Y. Fujimura,et al.  The 67kDa laminin receptor as a primary determinant of anti-allergic effects of O-methylated EGCG. , 2007, Biochemical and biophysical research communications.

[61]  M. Dembo,et al.  Cell movement is guided by the rigidity of the substrate. , 2000, Biophysical journal.

[62]  M. Textor,et al.  Optical grating coupler biosensors. , 2002, Biomaterials.

[63]  B. Péter,et al.  Biophysical characteristics of proteins and living cells exposed to the green tea polyphenol epigallocatechin-3-gallate (EGCg): review of recent advances from molecular mechanisms to nanomedicine and clinical trials , 2016, European Biophysics Journal.

[64]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[65]  Andreas Janshoff,et al.  Cell Adhesion Monitoring Using Substrate-Integrated Sensors , 2010 .

[66]  Robert Horvath,et al.  Sample handling in surface sensitive chemical and biological sensing: a practical review of basic fluidics and analyte transport. , 2014, Advances in colloid and interface science.

[67]  H. Mukhtar,et al.  Tea polyphenols: prevention of cancer and optimizing health. , 2000, The American journal of clinical nutrition.

[68]  Janos Vörös,et al.  RGD-grafted poly-L-lysine-graft-(polyethylene glycol) copolymers block non-specific protein adsorption while promoting cell adhesion. , 2003, Biotechnology and bioengineering.

[69]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[70]  R. Horváth,et al.  Multidepth screening of living cells using optical waveguides. , 2008, Biosensors & bioelectronics.