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Property Changes of Collagen Scaffolds Crosslinked by Traut’s reagent and Sulfo-SMCC
EAO Online Library. LI Y. Oct 9, 2018; 232618; P-BR-69
Yiming LI
Yiming LI
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Collagen scaffolds are frequently employed for applications in regenerative medicine. However, fast degradation rate and poor mechanical properties limited their application. Since collagen has many functional groups, moderate crosslinking modifcations would help to improve its properties. In previous studies, we adopted Traut’s reagent and Sulfo-SMCC to covalently combine cytokines on scaffolds and found that it could construct crosslinks of collagen scaffolds.**In order to evaluate the efficiency and bio-safety of Traut’s reagent and Sulfo-SMCC crosslinkers, we constructed collagen scaffolds with different extents of crosslinking by adjusting different ratios of these two reagents.**The collagen membrane purchased from Zhenghai Biotechnology was punched into standard size (diameter 6.4 mm, thickness of approximately 0.5 mm). The amino groups on the standard size of collagen scaffold were measured by NHN method. By adjusting ratios of Sulfo-SMCC and Traut’s reagent, collagen scaffolds of different extent of crosslinking were constructed and groups of significant difference were selected. The surface of collagen scaffolds were observed by scanning electron microscope, the structure was assessed by fourier transform infrared spectroscopy, in vitro degradation rate was measured by collagenase assay, cytocompatibility was evaluated by co-culturing HUVEC with collagen scaffolds or their extractions. After embedding in the subcutaneous pockets of SD rats for 7 days, the diameter of collagen scaffolds left were measured respectively. HE staining was performed to assess their cellularization.**By adjusting concentrations of Sulfo-SMCC, four different extent of crosslinking groups were selected. Scanning electron microscope showed collagen scaffolds became compact and the pores were more uniform and smooth after crosslinking. The Fourier transform infrared spectroscopy showed the structure was not altered. Cytocompatibility studies indicated the crosslinking procedure did not has toxicity. In vitro biodegradation studies showed with the increase of extent of crosslinking, the degradation resistance increased as well, as with in vivo studies. The SD rats subcutaneous pocket studies showed cellularization of crosslinked scaffolds were improved, which is related to the increasing degree of crosslinking. However, biological capacities of collagen scaffolds had no obvious changes when the inner-crossliking reaching an equilibration.**Using appropriate Traut’s reagent and Sulfo-SMCC is a safe and effective method to construct crosslinks within collagen scaffolds. This modification provides a new approach to improve biological capacities of naturally derived collagen scaffolds for potential use in tissue regeneration and wound healing.**
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