Preparation Of Sulfate Polysaccharide/typeⅠCollagen Scaffold And Its Application In Skin Regeneration

Objective: in order to elucidate the chemical structures and bioactiviity of sulfatepolysaccharides, the sulfate polysaccharides were purified and isolated, and thencomposed with typeⅠcollagen for regenerative scaffolds for the evaluation of theskin regeneration application.Methods: Two purified ingredients （F-2and F-3） were obtained from crudepolysaccharides by TCA-n-butyl alcohol and DEAE-52cellulose resin, and theirfundamental structures (F-2and F-3) were identified by multiple methods includingmetal ion content (ICP-MS), monosaccharide composition analysis (IC), elementalanalysis (EA), molecular weight (GPC), NMR, FT-IR; Carrageenan was purified byusing8%KCl, and its structure was tested by IC. Carrageenan simulated theexpressions of RAW264.7cells and promoted the proliferation of3T3cells by MTT.Type Ⅰ collagen/sulfate polysaccharides scaffolds were prepared successfully bysolvent-casting method. The morphology and aperture size of the scaffolds wasobserved by scanning electron microscopy; water absorption ratio and the in vitrodegradation characterization were performed;3T3cells adhesion on scaffolds wasobserved by SEM. The typeⅠcollagen/sulfate polysaccharides scaffolds were used toheal mice skin wound and its effect in process of skin regeneration was assessed. Theskin wound healing of mice was observed by HE staining.Results: the results showed that F-2was a complex structure of sulfatepolysaccharide with a molecular weight of2.65×104and18.43%sulfate groups. Itmainly included fucose, and also accompanied by galactose, a small amont ofrhamnose, xylose and glucose component. F-3was a complex structure of sulfatepolysaccharide with a molecular weight of2.95×105and31.53%sulfate groups. Itmainly included the fucose, and also accompanied by galactose, a small amont ofrhamnose, xylose and glucose component. The specific structural features of F-2andF-3were mainly composed by α-(1,3)-the fucose, also existed a small amount ofα-(1,4)-the fucose; side chains included massive1,3-linked,1,6-linked galactose,1,6-linked glucose, little β-D-xylose and rhamnose; the fucose sulfate-substituted had mainly2,3disubstituted location and2-substituted location, small2,4disubstitutedlocation in F-2; the fucose sulfate-substituted had mainly2,4disubstituted location,small2,4disubstituted location in F-3.The three types of carrageenans promoted theproliferation of3T3cell and had no effect on cell morphology; and the higher contentof sulfate groups, and better proliferation effect.Type I collagen/fucoidan and Type Icollagen/κ-carrageenan scaffolds were prepared successfully by solvent-castingmethod. These scaffolds had good biocompatibility, included appropriate pore size,porous network structure, strong water absorption, the appropriate rate of degradationand non-cytotoxic.The trauma repair results in mice experiments showed that FUC/Cand κ-C/C scaffolds were same effects with collagen sponge scaffold in promoting thehealing of skin wounds. These results demonstrate FUC/C and κ-C/C scaffolds havepromises in skin repair.