Atomic Force Microscopy surface nanocharacterization of UV-irradiated collagen thin films

Collagen, the most abundant protein in mammals, is a basic component of the extracellular matrix and due to its unique properties it is widely used as biomaterial, scaffold and culture substrate for cell and tissue regeneration studies. Due to human skin chronic exposure to sun light and since UV rays are used as sterilizing and cross-linking methods the clarification of the UV light-collagen interactions are very crucial. Moreover, since the majority of the biological reactions occur on surfaces or interfaces the influence of UV light on the surface of collagen-based materials attracts the scientific interest, especially in the biomaterials science. Surface-nanoscale characterization could be performed with Atomic Force Microscopy (AFM), which is a powerful tool and offers quantitative and qualitative information. Its ability of high resolution imaging and non-destructive characterization makes it very attractive for biological samples investigation. The aim of this paper was to determine the surface properties and alterations of collagen thin films after UV-irradiations using AFM techniques. Furthermore, it was aimed to investigate the possible different influence on the surface when the collagen solution or the thin films were irradiated. In this paper topographic AFM images were acquired from thin films, formed from both irradiated and non-irradiated collagen solutions, with spin coating procedure. The results demonstrated that the UV irradiation have different results when it is applied in the collagen solution or in the film after the spin coating methodology. For short irradiation times (<;120 min) UV caused only rather small changes in the morphology of the studied films although fluorescence and absorption studies confirmed collagen photodegradation. The surface roughness and topography altered after 3 and 7 hours, respectively, while the fibrous structure was completely destroyed after 15 hours. Surface roughness of the films depends on whether the solution was irradiated or the film and on the time irradiation. The fully clarification of the role of the UV light on collagen thin films will enable the proper design and control of collagen based nanobiomaterials with appropriate and improved surface properties.

[1]  L. Moldovan,et al.  Collagen–based scaffolds for skin tissue engineering , 2011, Journal of medicine and life.

[2]  C. Remacle,et al.  Supramolecular assemblies of adsorbed collagen affect the adhesion of endothelial cells. , 2006, Journal of biomedical materials research. Part A.

[3]  B. Brodsky,et al.  UV damage of collagen: insights from model collagen peptides. , 2012, Biopolymers.

[4]  I. Ionitaa,et al.  COLLAGEN FLUORESCENCE MEASUREMENTS ON NANOSILVER TREATED LEATHER , 2010 .

[5]  A. Sionkowska,et al.  Collagen fibrils in UV irradiated poly(vinyl pyrrolidone) films , 2008 .

[6]  John T Elliott,et al.  Cell response to matrix mechanics: focus on collagen. , 2009, Biochimica et biophysica acta.

[7]  A. Sionkowska Collagen Based Materials for Biomedical Applications: Preparation and Properties , 2012 .

[8]  A. Sionkowska,et al.  Surface properties of UV-irradiated poly(vinyl alcohol) films containing small amount of collagen , 2009 .

[9]  H. Shibata,et al.  Effect of ultraviolet radiation on photodegradation of collagen , 1999 .

[10]  N. Gibbs,et al.  Low‐dose ultraviolet radiation selectively degrades chromophore‐rich extracellular matrix components , 2010, The Journal of pathology.

[11]  D. Kaplan,et al.  Guide to collagen characterization for biomaterial studies. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[12]  A. Stylianou,et al.  Atomic Force Microscopy Imaging of the Nanoscale Assembly of Type I Collagen on Controlled Polystyrene Particles Surfaces , 2011 .

[13]  A. Stylianou,et al.  Atomic force microscopy quantitative and qualitative nanoscale characterization of collagen thin films , 2012 .

[14]  A. Sionkowska,et al.  UV-vis and FT-IR spectra of ultraviolet irradiated collagen in the presence of antioxidant ascorbic acid. , 2010, Ecotoxicology and environmental safety.

[15]  C. Gerber,et al.  Exogenous collagen cross‐linking recovers tendon functional integrity in an experimental model of partial tear , 2012, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  Kostas Politopoulos,et al.  Combined information from AFM imaging and SHG signal analysis of collagen thin films , 2011, Biomed. Signal Process. Control..

[17]  A. Stylianou,et al.  Combined SHG signal information with AFM imaging to assess conformational changes in collagen , 2009, 2009 9th International Conference on Information Technology and Applications in Biomedicine.

[18]  B. Nair,et al.  Effect of UV irradiation on the physicochemical properties of collagen stabilized using aldehydes , 2007 .

[19]  J. Gómez‐Herrero,et al.  WSXM: a software for scanning probe microscopy and a tool for nanotechnology. , 2007, The Review of scientific instruments.

[20]  P. So,et al.  Biologically active collagen-based scaffolds: advances in processing and characterization , 2010, Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences.

[21]  J. Hasenwinkel,et al.  Micropatterned agarose scaffolds covalently modified with collagen for culture of normal and neoplastic hepatocytes. , 2012, Journal of biomedical materials research. Part A.

[22]  Clair Baldock,et al.  Collagens at a glance , 2007, Journal of Cell Science.

[23]  Mitchel J Doktycz,et al.  Atomic force microscopy of biological samples. , 2010, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[24]  Simon Scheuring,et al.  Biological AFM: where we come from – where we are – where we may go , 2011, Journal of molecular recognition : JMR.

[25]  Yung Chang,et al.  Type I collagen structure modulates the behavior of osteoblast-like cells , 2010 .

[26]  T. Ramasami,et al.  Effect of UV irradiation on stabilized collagen: role of chromium(III). , 2008, Colloids and surfaces. B, Biointerfaces.

[27]  The Importance of Collagen Fibers in Vertebrate Biology , 2009 .

[28]  A. P. Gunning,et al.  Atomic Force Microscopy for Biologists , 1999 .

[29]  A. Sionkowska,et al.  Surface characterization of collagen/elastin based biomaterials for tissue regeneration , 2009 .

[30]  A. Stylianou,et al.  Surface Characterization of Collagen Films by Atomic Force Microscopy , 2010 .

[31]  Peter Fratzl,et al.  Collagen : structure and mechanics , 2008 .

[32]  I. Banerjee,et al.  Caprine (Goat) Collagen: A Potential Biomaterial for Skin Tissue Engineering , 2012, Journal of biomaterials science. Polymer edition.