Exploring the Limits of Cell Adhesion under Shear Stress within Physiological Conditions and beyond on a Chip

Cell adhesion processes are of ubiquitous importance for biomedical applications such as optimization of implant materials. Here, not only physiological conditions such as temperature or pH, but also topographical structures play crucial roles, as inflammatory reactions after surgery can diminish osseointegration. In this study, we systematically investigate cell adhesion under static, dynamic and physiologically relevant conditions employing a lab-on-a-chip system. We screen adhesion of the bone osteosarcoma cell line SaOs-2 on a titanium implant material for pH and temperature values in the physiological range and beyond, to explore the limits of cell adhesion, e.g., for feverish and acidic conditions. A detailed study of different surface roughness Rq gives insight into the correlation between the cells’ abilities to adhere and withstand shear flow and the topography of the substrates, finding a local optimum at Rq = 22 nm. We use shear stress induced by acoustic streaming to determine a measure for the ability of cell adhesion under an external force for various conditions. We find an optimum of cell adhesion for T = 37 °C and pH = 7.4 with decreasing cell adhesion outside the physiological range, especially for high T and low pH. We find constant detachment rates in the physiological regime, but this behavior tends to collapse at the limits of 41 °C and pH 4.

[1]  E. Stamhuis,et al.  PIVlab - Time-Resolved Digital Particle Image Velocimetry Tool for MATLAB , 2015 .

[2]  A. Wixforth,et al.  Acoustotaxis -in vitro stimulation in a wound healing assay employing surface acoustic waves. , 2016, Biomaterials science.

[3]  C. Oakley,et al.  The sequence of alignment of microtubules, focal contacts and actin filaments in fibroblasts spreading on smooth and grooved titanium substrata. , 1993, Journal of cell science.

[4]  A. Wixforth,et al.  Antibacterial metal ion release from diamond-like carbon modified surfaces for novel multifunctional implant materials , 2016 .

[5]  Shu Chien,et al.  Role of integrins in endothelial mechanosensing of shear stress. , 2002, Circulation research.

[6]  U von Stockar,et al.  Monitoring of temperature effects on animal cell metabolism in a packed bed process. , 2002, Biotechnology and bioengineering.

[7]  H. Mellor,et al.  Actin stress fibres , 2007, Journal of Cell Science.

[8]  W. Linnemans,et al.  The effects of hyperthermia on the cytoskeleton: a review. , 1996, International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group.

[9]  W. Pegios,et al.  Radiochemotherapy and hyperthermia in the treatment of rectal cancer. , 1998, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[10]  B D Boyan,et al.  Role of material surfaces in regulating bone and cartilage cell response. , 1996, Biomaterials.

[11]  J E Bailey,et al.  Effect of ammonium ion and extracellular pH on hybridoma cell metabolism and antibody production , 1990, Biotechnology and bioengineering.

[12]  J E Bailey,et al.  Influence of low temperature on productivity, proteome and protein phosphorylation of CHO cells. , 1999, Biotechnology and bioengineering.

[13]  P. Wust,et al.  The cellular and molecular basis of hyperthermia. , 2002, Critical reviews in oncology/hematology.

[14]  M. Gimbrone,et al.  Influence of hemodynamic forces on vascular endothelial function. In vitro studies of shear stress and pinocytosis in bovine aortic cells. , 1984, The Journal of clinical investigation.

[15]  M. J. Nine,et al.  Wear Debris Characterization and Corresponding Biological Response: Artificial Hip and Knee Joints , 2014, Materials.

[16]  R. Lang-Roth,et al.  Allgemeine und spezielle Pharmakologie und Toxikologie , 2009 .

[17]  Benjamin Geiger,et al.  Cell spreading and focal adhesion dynamics are regulated by spacing of integrin ligands. , 2007, Biophysical journal.

[18]  S. Kurtz,et al.  Projections of primary and revision hip and knee arthroplasty in the United States from 2005 to 2030. , 2007, The Journal of bone and joint surgery. American volume.

[19]  J. Zweier,et al.  pH dependence of neutrophil-endothelial cell adhesion and adhesion molecule expression. , 1996, The American journal of physiology.

[20]  William Thielicke,et al.  The flapping flight of birds: Analysis and application , 2014 .

[21]  E. Gerner,et al.  Heat shock-induced shedding of cell surface integrins in A549 human lung tumor cells in culture. , 1994, Experimental cell research.

[22]  I. Tannock,et al.  Acid pH in tumors and its potential for therapeutic exploitation. , 1989, Cancer research.

[23]  K. Shadan,et al.  Available online: , 2012 .

[24]  D. Deligianni,et al.  Effect of surface roughness of hydroxyapatite on human bone marrow cell adhesion, proliferation, differentiation and detachment strength. , 2001, Biomaterials.

[25]  A F von Recum,et al.  Fibroblast anchorage to microtextured surfaces. , 1993, Journal of biomedical materials research.

[26]  L. Weiss Cell contact phenomena , 2007, In Vitro.

[27]  C. Murray,et al.  A review of dental implants and infection. , 2009, The Journal of hospital infection.

[28]  Amelia Ahmad Khalili,et al.  A Review of Cell Adhesion Studies for Biomedical and Biological Applications , 2015, International journal of molecular sciences.

[29]  Achim Wixforth,et al.  A novel tool for dynamic cell adhesion studies--the De-Adhesion Number Investigator DANI. , 2014, Lab on a chip.

[30]  B. Nebe,et al.  Mechanical Stressing of Integrin Receptors Induces Enhanced Tyrosine Phosphorylation of Cytoskeletally Anchored Proteins* , 1998, The Journal of Biological Chemistry.

[31]  Achim Wixforth,et al.  A surface acoustic wave-driven micropump for particle uptake investigation under physiological flow conditions in very small volumes , 2015, Beilstein journal of nanotechnology.

[32]  N. Ishiguro,et al.  Osteoclast induction from bone marrow cells is due to pro-inflammatory mediators from macrophages exposed to polyethylene particles: a possible mechanism of osteolysis in failed THA. , 2001, Journal of biomedical materials research.

[33]  Jerry Westerweel,et al.  Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines. , 2009, Lab on a chip.

[34]  Stephen C Cowin,et al.  Blood and interstitial flow in the hierarchical pore space architecture of bone tissue. , 2015, Journal of biomechanics.

[35]  C. Turner,et al.  Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. , 1988, Annual review of cell biology.

[36]  R. Skalak,et al.  Design and construction of a linear shear stress flow chamber , 2006, Annals of Biomedical Engineering.

[37]  R. Spier,et al.  The adhesion of animal cells to surfaces: The measurement of critical surface shear stress permitting attachment or causing detachment , 1985 .

[38]  J. M. Engasser,et al.  Determination of cell lysis and death kinetics in continuous hybridoma cultures from the measurement of lactate dehydrogenase release , 2004, Cytotechnology.

[39]  William Thielicke,et al.  PIVlab – Towards User-friendly, Affordable and Accurate Digital Particle Image Velocimetry in MATLAB , 2014 .

[40]  Maxence Bigerelle,et al.  Qualitative and quantitative study of human osteoblast adhesion on materials with various surface roughnesses. , 2000, Journal of biomedical materials research.

[41]  A. Konings,et al.  Role of membrane lipids and membrane fluidity in thermosensitivity and thermotolerance of mammalian cells. , 1985, Radiation research.

[42]  A. Wixforth,et al.  Flow patterns and transport in Rayleigh surface acoustic wave streaming: combined finite element method and raytracing numerics versus experiments , 2008, IEEE Transactions on Ultrasonics, Ferroelectrics and Frequency Control.

[43]  Her-Hsiung Huang,et al.  Effect of surface roughness of ground titanium on initial cell adhesion. , 2004, Biomolecular engineering.

[44]  A F von Recum,et al.  Quantitative analysis of cell proliferation and orientation on substrata with uniform parallel surface micro-grooves. , 1996, Biomaterials.

[45]  P. Charette,et al.  Cell detachment and label-free cell sorting using modulated surface acoustic waves (SAWs) in droplet-based microfluidics. , 2014, Lab on a chip.

[46]  A. Punnia-Moorthy Evaluation of pH changes in inflammation of the subcutaneous air pouch lining in the rat, induced by carrageenan, dextran and Staphylococcus aureus. , 1987, Journal of oral pathology.

[47]  K. Anselme,et al.  Osteoblast adhesion on biomaterials. , 2000, Biomaterials.

[48]  W. Moolenaar Effects of growth factors on intracellular pH regulation. , 1986, Annual review of physiology.

[49]  M. Laroche Intraosseous circulation from physiology to disease. , 2002, Joint, bone, spine : revue du rhumatisme.

[50]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.