Advertisement

Electric Cell-Substrate Impedance Sensing as a Screening Tool for Wound Healing Agents

  • Cheuk Lun Liu
  • Jacqueline Chor Wing Tam
  • Andrew J. Sanders
  • David G. Jiang
  • Chun Hay Ko
  • Kwok Pui Fung
  • Ping Chung Leung
  • Keith G. Harding
  • Wen G. Jiang
  • Clara Bik San Lau
Chapter
Part of the Cancer Metastasis - Biology and Treatment book series (CMBT, volume 17)

Abstract

As one of the earliest matured techniques for in vitro cell migration study in wound healing, traditional scratch assay has been routinely utilized due to its simplicity of setup in cell culture. However, with the emerging needs of acquiring high sensitivity and achieving high-throughput in cell behavior study, researchers in life science started to apply electric cell-substrate impedance sensing (ECIS) technology, with its automated real-time impedance monitoring and standard electrical wounding, in various study fields. Nonetheless, not much information is available regarding the application of ECIS in the screening of wound healing agents. Here we first reviewed the applications of both traditional scratch assay and ECIS model in wound healing. We further used a herbal formula NF3 (comprising of individual herbs Astragali Radix and Rehmanniae Radix in the ratio of 2:1) which was previously shown to exhibit profound wound-healing effect in diabetic foot ulcer rat model, as an example for comparison of cell migration studies using both traditional scratch assay and ECIS model. To conclude, with its high sensitivity and efficiency, ECIS demonstrated its reliability as a tool for the screening of wound healing agents.

Keywords

Wound Healing Human Umbilical Vein Endothelial Cell HaCaT Cell Scratch Assay Activate Leukocyte Cell Adhesion Molecule 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgement

This study was supported by the University Grants Committee of the Hong Kong SAR under the Area of Excellence project “Chinese Medicine Research and Further Development” (Ref. No. AoE/B-10/01). We would also like to thank Hop Wai short-term research fellowship for the sponsorship of ECIS training in Cardiff University School of Medicine, Cardiff, United Kingdom.

References

  1. Ablin RJ, Kynaston HG, Mason MD, Jiang WG (2011) Prostate transglutaminase (TGase-4) antagonizes the anti-tumour action of MDA-7/IL-24 in prostate cancer. J Transl Med 9:49PubMedCrossRefGoogle Scholar
  2. Barrientos S, Stojadinovic O, Golinko MS, Brem H, Tomic-Canic M (2008) Growth factors and cytokines in wound healing. Wound Repair Regen 16:585–601PubMedCrossRefGoogle Scholar
  3. Betzen C, White R, Zehendner CM, Pietrowski E, Bender B, Luhmann HJ, Kuhlmann CR (2009) Oxidative stress upregulates the NMDA receptor on cerebrovascular endothelium. Free Radic Biol Med 47:1212–1220PubMedCrossRefGoogle Scholar
  4. Diegelmann RF, Evans MC (2004) Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci 9:283–289PubMedCrossRefGoogle Scholar
  5. Fan TPD, Jaggar R, Bicknell R (1995) Controlling the vasculature: angiogenesis, anti-angiogenesis and vascular targeting of gene therapy. Trends Pharmacol Sci 16:57–66PubMedCrossRefGoogle Scholar
  6. Gailit J, Clark RA (1996) Studies in vitro on the role of alpha v and beta 1 integrins in the adhesion of human dermal fibroblasts to provisional matrix proteins fibronectin, vitronectin, and fibrinogen. J Invest Dermatol 106:102–108PubMedCrossRefGoogle Scholar
  7. Gebäck T, Schulz MMP, Koumoutsakos P, Detmar M (2009) Tscratch: a novel and simple software tool for automated analysis of monolayer wound healing assays. Short Tech Rep 46:265–274Google Scholar
  8. Giaever I, Keese CR (1984) Monitoring fibroblast behavior in tissue culture with an applied electric field. Proc Natl Acad Sci U S A 81:3761–3764PubMedCrossRefGoogle Scholar
  9. Goetsch KP, Niesler CU (2011) Optimization of the scratch assay for in vitro skeletal muscle wound healing analysis. Anal Biochem 411:158–160PubMedCrossRefGoogle Scholar
  10. Grab DJ, Nyarko E, Nikolskaia OV, Kim YV, Dumler JS (2009) Human brain microvascular endothelial cell traversal by Borrelia burgdorferi requires calcium signaling. Clin Microbiol Infect 15:422–426PubMedCrossRefGoogle Scholar
  11. Grinnell F (1990) The activated keratinocyte: upregulation of cell adhesion and migration during wound healing. J Trauma 30:S144–S149PubMedCrossRefGoogle Scholar
  12. Grinnell F, Toda K, Takashima A (1987) Activation of keratinocyte fibronectin receptor function during cutaneous wound healing. J Cell Sci 8:199–209Google Scholar
  13. Heijink IH, Kies PM, Kauffman HF, Postma DS, Van Oosterhout AJ, Vellenga E (2007) Down-regulation of E-cadherin in human bronchial epithelial cells leads to epidermal growth factor receptor-dependent Th2 cell-promoting activity. J Immunol 178:7678–7685PubMedGoogle Scholar
  14. Heijink IH, Brandenburg SM, Noordhoek JA, Postma DS, Slebos DJ, van Oosterhout AJ (2010) Characterisation of cell adhesion in airway epithelial cell types using electric cell-substrate impedance sensing. Eur Respir J 35:894–903PubMedCrossRefGoogle Scholar
  15. Hsu CC, Tsai WC, Chen CP, Lu YM, Wang JS (2010) Effects of negative pressures on epithelial tight junctions and migration in wound healing. Am J Cell Physiol 299:C528–C534CrossRefGoogle Scholar
  16. Jiang WG, Ye L, Patel G, Harding KG (2010) Expression of WAVEs, the WASP (Wiskott-Aldrich syndrome protein) family of verprolin homologous proteins in human wound tissues and the biological influence on human keratinocytes. Wound Repair Regen 18:594–604PubMedCrossRefGoogle Scholar
  17. Kandasamy K, Choi CS, Kim S (2010) An efficient analysis of nanomaterial cytotoxicity based on bioimpedance. Nanotechnology 21:375501–375510PubMedCrossRefGoogle Scholar
  18. Keese CR, Wegener J, Walker SR, Giaever I (2004) Electrical wound-healing assay for cells in vitro. Proc Natl Acad Sci U S A 101:1554–1559PubMedCrossRefGoogle Scholar
  19. Lamalice L, Boeuf FL, Huot J (2007) Endothelial cell migration during angiogenesis. Circ Res 100:782–794PubMedCrossRefGoogle Scholar
  20. Lee WK, Torchalski B, Kohistani N, Thévenod F (2011) ABCB1 protects kidney proximal tubule cells against cadmium-induced apoptosis: roles of cadmium and ceramide transport. Toxicol Sci 121:343–356PubMedCrossRefGoogle Scholar
  21. Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2:2Google Scholar
  22. Lo CM, Keese CR, Giaever I (1993) Monitoring motion of confluent cells in tissue culture. Exp Cell Res 204:102–109PubMedCrossRefGoogle Scholar
  23. Moore E, Rawley O, Wood T, Galvin P (2009) Monitoring of cell growth in vitro using biochips packaged with indium tin oxide sensors. Sens Actuator B Chem 139:187–193CrossRefGoogle Scholar
  24. Oberringer M, Meins C, Bubel M, Pohlemann T (2007) A new in vitro wound model based on the co-culture of human dermal microvascular endothelial cells and human dermal fibroblasts. Biol Cell 99:197–207PubMedCrossRefGoogle Scholar
  25. Patani N, Douglas-Jones A, Mansel R, Jiang W, Mokbel K (2010) Tumour suppressor function of MDA-7/IL-24 in human breast cancer. Cancer Cell Int 10:29PubMedGoogle Scholar
  26. Ramirez-Icaza G, Mohammed KA, Nasreen N, Van Horn RD, Hardwick JA, Sanders KL, Tian J, Ramirez-Icaza C, Johnson MT, Antony VB (2004) Th2 cytokines IL-4 and IL-13 downregulate paxillin expression in bronchial airway epithelial cells. J Clin Immunol 24:426–434PubMedCrossRefGoogle Scholar
  27. Ramzato E, Martinotti S, Volante A, Mazzucco L, Burlando B (2010) Platelet lyzate modulates MMP-2 and MMP-9 expression, matrix deposition and cell-to-matrix adhesion in keratinocytes and fibroblasts. Exp Dermatol 20:308–313CrossRefGoogle Scholar
  28. Sanders AJ, Jiang DG, Jiang WG, Harding KG, Patel GK (2011) Activated leukocyte cell adhesion molecule impacts on clinical wound healing and inhibits HaCaT migration. Int Wound J 8:500–507PubMedCrossRefGoogle Scholar
  29. Sharma D, Wang J, Fu PP, Sharma S, Nagalingam A, Mells J, Handy J, Page AJ, Cohen C, Anania FA, Saxena NK (2010) Adiponectin antagonizes the oncogenic actions of leptin in hepatocellular carcinogenesis. Hepatology 52:1713–1722PubMedCrossRefGoogle Scholar
  30. Smita A, Hiroki U, Zheng CH, Cooper LA, Zhao J, Christopher R, Guan JL (2002) Regulation of focal adhesion kinase by a novel protein inhibitor FIP200. Mol Biol Cell 13:3178–3191CrossRefGoogle Scholar
  31. Tam JCW, Lau KM, Liu CL, To MH, Kwok HF, Lai KK, Lau CP, Ko CH, Leung PC, Fung KP, Lau CBS (2011) The in vivo and in vitro diabetic wound healing effects of a 2-herb formula and its mechanisms of action. J Ethnopharmacol 134:831–838PubMedCrossRefGoogle Scholar
  32. Tarantola M, Pietuch A, Schneider D, Rother J, Sunnick E, Rosman C, Pierrat S, Sönnichsen C, Wegener J, Janshoff A (2011) Toxicity of gold-nanoparticles: synergistic effects of shape and surface functionalization on micromotility of epithelial cells. Nanotoxicology 5:254–268PubMedCrossRefGoogle Scholar
  33. Von Wedel-Parlow M, Schrot S, Lemmen J, Treeratanapiboon L, Wegener J, Galla HJ (2011) Neutrophils cross the BBB primarily on transcellular pathways: an in vitro study. Brain Res 1367:62–76CrossRefGoogle Scholar
  34. Wiesner C, Pflüger M, Kopecky J, Stys D, Entler B, Lucas R, Hundsberger H, Schütt W (2008) Implementation of ECIS technology for the characterization of potential therapeutic drugs that promote wound-healing. GMS Krankenhhyg Interdiszip 3:Doc05Google Scholar
  35. Yarrow JC, Perlman ZE, Westwood NJ, Mitchison TJ (2004) A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods. BMC Biotechnol 4:21PubMedCrossRefGoogle Scholar
  36. Zantek ND, Kinch MS (2001) Analysis of cell migration. Method Cell Biol 63:549–559CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Cheuk Lun Liu
    • 1
    • 2
    • 3
  • Jacqueline Chor Wing Tam
    • 1
    • 2
  • Andrew J. Sanders
    • 4
  • David G. Jiang
    • 4
  • Chun Hay Ko
    • 1
    • 2
  • Kwok Pui Fung
    • 1
    • 2
    • 3
  • Ping Chung Leung
    • 1
    • 2
  • Keith G. Harding
    • 5
  • Wen G. Jiang
    • 4
  • Clara Bik San Lau
    • 1
    • 2
  1. 1.Institute of Chinese MedicineThe Chinese University of Hong KongShatinHong Kong
  2. 2.State Key Laboratory of Phytochemistry and Plant Resources in West ChinaThe Chinese University of Hong KongShatinHong Kong
  3. 3.School of Biomedical SciencesThe Chinese University of Hong KongShatinHong Kong
  4. 4.Metastasis and Angiogenesis Research GroupCardiff University School of MedicineCardiffUK
  5. 5.Department of Dermatology and Wound HealingCardiff University School of MedicineCardiffUK

Personalised recommendations