Matrices comprising a modified polysaccharide
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example 1
Modification of Agarose
[0130]Agarose type I was obtained from Calbiochem. (2,2,6,6-Tetramethylpiperidin-1-yl)oxyl (TEMPO), NaOCl, NaBH4, NaBr, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 2-(N-morpholino)ethanesulfonic acid (MES) buffer were obtained from Sigma Aldrich and used as received. Solution of 0.5 M NaOH as well as solution of 5 M HCl were freshly prepared every three-month. Ethanol technical grade was used without any further purification. Deionized water was used for non-sterile synthesis.
[0131]Agarose was modified under sterile conditions: all the chemicals were dissolved in autoclaved water and filtered with a 0.2 μm filter. All the glassware was autoclaved and the reaction was conducted under a laminar flow. Agarose (1 g) was autoclaved in MilliQ water. Autoclaved agarose was poured into a 3 necked round bottom flask, which was used as a reactor. A mechanical stirrer was adapted to one of the neck. A pH-meter was adapted another neck of the round bottom flas...
example 2
Modification of K-Carrageenan
[0132]K-carrageenan was obtained from Sigma Aldrich and used as such. K-carrageenan was objected to a TEMPO-mediated oxidation, according to the synthetic protocol of Example 1.
[0133]The resulting modified k-carrageenan was dialyzed and freeze-dried, as specified in Example 1 above.
example 3
Blending of Modified Agarose with Unmodified Agarose
[0134]Blends of modified agarose and native agarose in different proportions were prepared. The resulting blends were studied using CD spectrometry. The obtained results indicate that the blending of two polysaccharides lead to the same change in tertiary structure as the modified polysaccharide. This suggests that these two polysaccharides are miscible and can be used for engineering new matrices.
[0135]The rheology studies show a specific behavior of the gel. Indeed the blended gels have a higher shear modulus than the unmodified gel of agarose. These results suggest that the organization of the modified chains with the unmodified chains follows a new mechanism, that is different from modified agarose.
[0136]The ESEM image illustrates the structure of the gels and reveals a different organization of the fibers than for the unmodified agarose. As well for the surface roughness, the roughness of the blended gels is not dependent of t...
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