The process of EDC-NHS Cross-linking of reconstituted collagen fibres increases collagen fibrillar order and alignment.

We describe the production of collagen fibre bundles through a multi-strand, semi-continuous extrusion process. Cross-linking using an EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), NHS (N-hydroxysuccinimide) combination was considered. Atomic Force Microscopy (AFM) and Raman spectroscopy focused on how cross-linking affected the collagen fibrillar structure. In the cross-linked fibres, a clear fibrillar structure comparable to native collagen was observed which was not observed in the non-cross-linked fibre. The amide III doublet in the Raman spectra provided additional evidence of alignment in the cross-linked fibres. Raman spectroscopy also indicated no residual polyethylene glycol (from the fibre forming buffer) or water in any of the fibres.

[1]  I. Yannas,et al.  Recent advances in tissue synthesis in vivo by use of collagen-glycosaminoglycan copolymers. , 1996, Biomaterials.

[2]  F H Silver,et al.  Self-assembly of collagen fibers. Influence of fibrillar alignment and decorin on mechanical properties. , 1997, Biophysical journal.

[3]  F H Silver,et al.  Collagen fibres with improved strength for the repair of soft tissue injuries. , 1994, Biomaterials.

[4]  R. Cameron,et al.  Synthetic collagen fascicles for the regeneration of tendon tissue. , 2012, Acta biomaterialia.

[5]  N. Timofeeva,et al.  Observations of multiscale, stress-induced changes of collagen orientation in tendon by polarized Raman spectroscopy. , 2011, Biomacromolecules.

[6]  Kye-Yong Song,et al.  Reinforced bioartificial dermis constructed with collagen threads , 2008 .

[7]  R. G. Paul,et al.  The influence of a natural cross-linking agent (Myrica rubra) on the properties of extruded collagen fibres for tissue engineering applications , 2010 .

[8]  F H Silver,et al.  Mechanical properties of collagen fibres: a comparison of reconstituted and rat tail tendon fibres. , 1989, Biomaterials.

[9]  C. Hadjur,et al.  In vivo chemical investigation of human skin using a confocal Raman fiber optic microprobe. , 2005, Journal of biomedical optics.

[10]  R. Stark,et al.  Anisotropic Raman scattering in collagen bundles. , 2010, Optics letters.

[11]  D A Parry,et al.  Analysis of the primary structure of collagen for the origins of molecular packing. , 1973, Journal of molecular biology.

[12]  A. Vergara,et al.  Correlation between Raman and X-ray crystallography data of (Pro-Pro-Gly)10. , 2008, Biophysical chemistry.

[13]  R. Cameron,et al.  Effect of fiber crosslinking on collagen-fiber reinforced collagen-chondroitin-6-sulfate materials for regenerating load-bearing soft tissues. , 2013, Journal of biomedical materials research. Part A.

[14]  Alois Bonifacio,et al.  Effects of sample orientation in Raman microspectroscopy of collagen fibers and their impact on the interpretation of the amide III band , 2010 .

[15]  S. Lazare,et al.  Spectroscopic study of a KrF excimer laser treated surface of the thin collagen films , 2007 .

[16]  J. Revel,et al.  Subfibrillar structure of type I collagen observed by atomic force microscopy. , 1993, Biophysical journal.

[17]  E. Chaikof,et al.  D-periodic collagen-mimetic microfibers. , 2007, Journal of the American Chemical Society.

[18]  E. J. Miller,et al.  Effect of physical crosslinking methods on collagen-fiber durability in proteolytic solutions. , 1996, Journal of biomedical materials research.

[19]  R. G. Paul,et al.  Post-self-assembly experimentation on extruded collagen fibres for tissue engineering applications. , 2008, Acta biomaterialia.

[20]  D. Chau,et al.  The cellular response to transglutaminase-cross-linked collagen. , 2005, Biomaterials.

[21]  Umut A. Gurkan,et al.  An electrochemical fabrication process for the assembly of anisotropically oriented collagen bundles. , 2008, Biomaterials.

[22]  Dimitrios I Zeugolis,et al.  Cross-linking of extruded collagen fibers--a biomimetic three-dimensional scaffold for tissue engineering applications. , 2009, Journal of biomedical materials research. Part A.

[23]  F. O'Brien,et al.  The effect of dehydrothermal treatment on the mechanical and structural properties of collagen-GAG scaffolds. , 2009, Journal of biomedical materials research. Part A.

[24]  A. Stemmer,et al.  Mechanical response of individual collagen fibrils in loaded tendon as measured by atomic force microscopy. , 2011, Journal of structural biology.

[25]  H. Screen,et al.  The micro-structural strain response of tendon , 2007 .

[26]  Ws. Rasband.  ImageJ, U.S. National Institutes of Health, Bethesda, Maryland, USA , 2011 .

[27]  S. Andreadis,et al.  Crosslinking of discrete self-assembled collagen threads: Effects on mechanical strength and cell-matrix interactions. , 2007, Journal of biomedical materials research. Part A.

[28]  Neil Rushton,et al.  Collagen fibre implant for tendon and ligament biological augmentation. In vivo study in an ovine model , 2013, Knee Surgery, Sports Traumatology, Arthroscopy.

[29]  J. Koenig,et al.  Raman scattering of collagen, gelatin, and elastin , 1975, Biopolymers.

[30]  George Z. Papageorgiou,et al.  Characterization of the distribution, polymorphism, and stability of nimodipine in its solid dispersions in polyethylene glycol by micro-Raman spectroscopy and powder x-ray diffraction , 2007, The AAPS Journal.

[31]  D L Bader,et al.  Determination of molecular changes in soft tissues under strain using laser Raman microscopy. , 2000, Journal of biomechanics.

[32]  B. Cwalina,et al.  Atomic force microscopy investigation of chemically stabilized pericardium tissue , 2005, The European physical journal. E, Soft matter.

[33]  M. Dunn,et al.  Functional evaluation of collagen fiber scaffolds for ACL reconstruction: cyclic loading in proteolytic enzyme solutions. , 2004, Journal of biomedical materials research. Part A.

[34]  F. Sommer,et al.  Atomic force microscopy study of the collagen fibre structure , 1994, Biology of the cell.

[35]  F. Silver,et al.  A self-assembled collagen scaffold suitable for use in soft and hard tissue replacement , 1995 .

[36]  D. Speer,et al.  Effect of tanning agent on tissue reaction to tissue implanted collagen sponge. , 1983, The Journal of surgical research.