15 chi and gel based cryogels were prepared using

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15CHI and Gel based cryogels were prepared using polymeric and monomeric nontoxic475cross-linking agents such as dextran dialdehyde and tetramethylpropane. Gel-DDA and CHI-476DDA macroporous cryogels were prepared in two-steps. For the first time TMP cross-linker477withoutuseoftoxicreducingagentwasappliedforthecryogelpreparation.CHI-DDA478cryogelswithoutreduction,butwithfreezedryingshowed~2higherdegradationdegree479compared to CHI-DDAtreated with borohydride. The surface of CHIcryogelswith DDA480showed higher zeta potential compared to the CHI-TMP. CHI and Gel containing cryogels481cross-linkedbyglutaraldehydehadcellviabilityapproximately40%,whilecryogels482synthesizedwiththeproposedmethodsshowedgoodbiocompatibility(~80%)and483proliferation efficiency of fibroblast cellsin vitro. Using confocal observer SD Carl Zeiss the4843D images of cells attached to the cryogel surface were obtained and their proliferation into485thedepthofthematerialwasshown.Suggestedcryogelswidenalibraryofavailable486scaffolds based on natural polymers which can be used for various purposes of regenerative487medicine.488489Conflicts of interest490There are no conflicts to declare491Supplementary materials492See supplementary materials related to this article.493Acknowledgements494The authors are grateful to the Department of Biotechnology and personally Emeritus495Prof. Bo Mattiasson of Lund University for providing access to facilities and chemicals. DB496gratefully acknowledges that the European Union’s Horizon 2020 Research and Innovation497Programme under the Marie Skłodowska-Curie Fellowship grant agreement 701289,498supported time when writing this paper. Authors are grateful to Sholpan Kauanova for the499help with the confocal imaging and image processing.500501References502[1]C.Y. Kuo, C.H. Chen, C.Y. Hsiao, J.P. Chen, Incorporation of chitosan in biomimetic503gelatin/chondroitin-6-sulfate/hyaluronancryogelforcartilagetissueengineering,504Carbohydr. Polym. 117 (2015) 722–730. doi:10.1016/j.carbpol.2014.10.056.505[2]E.Jain,A.Kumar,Disposablepolymericcryogelbioreactormatrixfortherapeutic506
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pH, buffer solution, Gelatin

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