Cryo-copolymerization preparation of dextran-hyaluronate based supermacroporous cryogel scaffolds for tissue engineering applications

Abstract

Dextran-hyaluronate (Dex-HA) based supermacroporous cryogel scaffolds for soft tissue engineering were prepared by free radical cryo-copolymerization of aqueous solutions containing the dextran methacrylate (Dex-MA) and hyaluronate methacrylate (HA-MA) at various macromonomer concentrations under the freezing condition. It was observed that the suitable total concentration of macromonomers for the preparation of Dex-HA cryogel scaffold with satisfied properties was 5% (w/w) at the HA-MA concentration of 1% (w/v), which was then used to produce the test scaffold. The obtained cryogel scaffold with 5% (w/w) macromonomer solution had high water permeability (5.1 × 10^-12 m²) and high porosity (92.4%). The pore diameter examined by scanning electron microscopy (SEM) was in a broad range of 50–135 µm with the mean pore diameter of 91 µm. Furthermore, the cryogel scaffold also had good elastic nature with the elastic modulus of 17.47±1.44 kPa. The culture of 3T3-L1 preadipocyte within the scaffold was investigated and observed by SEM. Cells clustered on the pore walls and grew inside the scaffold indicating the Dex-HA cryogel scaffold could be a promising porous biomaterial for applications in tissue engineering.

Keywords

cryogel scaffold / tissue engineering / dextran / hyaluronate / 3T3-L1 preadipocyte

References

[1] Malafaya P B, Silva G A, Reis R L. Natural-origin polymers as carriers and scaffolds for biomolecules and cell delivery in tissue engineering applications. Advanced Drug Delivery Reviews, 2007, 59(4-5): 207–233

[2] Liu C, Xia Z, Czernuszka J. Design and development of three-dimensional scaffolds for tissue engineering. Chemical Engineering Research & Design, 2007, 85(7): 1051–1064

[3] van Vlierberghe S, Cnudde V, Dubruel P, Masschaele B, Cosijns A, De Paepe I, Jacobs P J S, van Hoorebeke L, Remon J P, Schacht E. Porous gelatin hydrogels: cryogenic formation and structure analysis. Biomacromolecules, 2007, 8(2): 331–337

[4] Kathuria N, Tripathi A, Kar K, Kumar A. Synthesis and characterization of elastic and macroporous chitosan-gelatin cryogels for tissue engineering. Acta Biomaterialia, 2009, 5(1): 406–418

[5] Lévesque S G, Lim R M, Shoichet M S. Macroporous interconnected dextran scaffolds of controlled porosity for tissue-engineering applications. Biomaterials, 2005, 26(35): 7436–7446

[6] Möller S, Weisser J, Bischoff S, Schnabelrauch M. Dextran and hyaluronan methacrylate based hydrogels as matrices for soft tissue reconstruction. Biomolecular Engineering, 2007, 24(5): 496–504

[7] Autissier A, Visage C L, Pouzet C, Chaubet F, Letourneur D. Fabrication of porous polysaccharide-based scaffolds using a combined freeze-drying/cross-linking process. Acta Biomaterialia, 2010, 6(9): 3640–3648

[8] Davidenko N, Campbell J J, Thian E S, Watson C J, Cameron R E. Collagen-hyaluronic acid scaffolds for adipose tissue engineering. Acta Biomaterialia, 2010, 6(10): 3957–3968

[9] Ibrahim S, Kothapalli C R, Kang Q K, Ramamurthi A. Characterization of glycidyl ethacrylate-crosslinked hyaluronan hydrogel scaffolds incorporating elastogenic hyaluronan oligomers. Acta Biomaterialia, 2010, 7(3): 653–665

[10] Bloch K, Lozinsky V I, Galaev I Y, Yavriyanz K, Vorobeychik M, Azarov D, Damshkaln L G, Mattiasson B, Vardi P. Functional activity of insulinoma cells (INS-1E) and pancreatic islets cultured in agarose cryogel sponges. Journal of Biomedical Materials Research, 2005, 75(4): 802–809

[11] Plieva F M, Oknianska A, Degerman E, Galaev I Y, Mattiasson B. Novel supermacroporous dextran gels. Journal of Biomaterials Science. Polymer Edition, 2006, 17(10): 1075–1092

[12] Plieva F M, Xiao H T, Galaev I Y, Bergenståhl B, Mattiasson B. Macroporous elastic polyacrylamide gels prepared at subzero temperatures: control of porous structure. Journal of Materials Chemistry, 2006, 16(41): 4065–4073

[13] Plieva F M, Galaev I Y, Mattiasson B. Macroporous gels prepared at subzero temperatures as novel materials for chromatography of particulate-containing fluids and cell culture applications. Journal of Separation Science, 2007, 30(11): 1657–1671

[14] Dainiak M B, Allan I U, Savina I N, Cornelio L, James E S, James S L, Mikhalovsky S V, Jungvid H, Galaev I Y. Gelatin-fibrinogen cryogel dermal matrices for wound repair: preparation, optimisation and in vitro study. Biomaterials, 2010, 31(1): 67–76

[15] Lozinsky V I. Cryogels on the basis of natural and synthetic polymers: preparation, properties and applications. Russian Chemical Reviews, 2002, 71(6): 489–511

[16] Lozinsky V I. Polymeric cryogels as a new family of macroporous and supermacroporous materials for biotechnological purposes. Russian Chemical Bulletin, 2008, 57(5): 1015–1032

[17] Yao K J, Yun J X, Shen S C, Wang L H, He X J, Yu X M. Characterization of a novel continuous supermacroporous monolithic cryogel embedded with nanoparticles for protein chromatography. Journal of Chromatography. A, 2006, 1109(1): 103–110

[18] Yun J X, Shen S C, Chen F, Yao K J. One-step isolation of adenosine triphosphate from crude fermentation broth of Saccharomyces cerevisiae by anion-exchange chromatography using supermacroporous cryogel. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 2007, 860(1): 57–62

[19] Plieva F M, Galaev I, Noppe W, Mattiasson B. Cryogel applications in microbiology. Trends in Microbiology, 2008, 16(11): 543–551

[20] Nilsang S, Nehru V, Plieva F M, Nandakumar K S, Rakshit S K, Holmdahl R, Mattiasson B, Kuma A. Three-dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles. Biotechnology Progress, 2008, 24(5): 1122–1131

[21] Kirsebom H, Aguilar M R, Roman J S, Fernandez M, Prieto M A, Bondar B. Macroporous scaffolds based on chitosan and bioactive molecules. Journal of Bioactive and Compatible Polymers, 2007, 22(6): 621–636

[22] Cadéel J A, van Luyn M J A, Brouwer L A, Plantinga J A, van Wachem P B, de Groot C J, den Otter W, Hennink W E. In vivo biocompatibility of dextran-based hydrogels. Journal of Biomedical Materials Research, 2000, 50(3): 397–404

[23] de Groot C J, van Luyn M J A, van Dijk-Wolthuis W N E, Cadéel J A, Plantinga J A, den Otter W, Hennink W E. In vitro biocompatibility of biodegradable dextran-based hydrogels tested with human fibroblasts. Biomaterials, 2001, 22(11): 1197–1203

[24] Liu Y X, Chan-Park M B. Hydrogel based on interpenetrating polymer networks of dextran and gelatin for vascular tissue engineering. Biomaterials, 2009, 30(2): 196–207

[25] van Dijk-Wolthuis W N E, Franssen O, Talsma H, van Steenbergen M J, Kettenes-van den Bosch J J, Hennink W E. Synthesis, characterization, and polymerization of glycidyl methacrylate derivatized dextran. Macromolecules, 1995, 28(18): 6317–6322

[26] van Dijk-Wolthuis W N E, Kettenes-van den Bosch J J, van der Kerk-van Hoof A, Hennink W E. Reaction of dextran with glycidyl methacrylate: an unexpected transesterification. Macromolecules, 1997, 30(11): 3411–3413

[27] Yao K J, Shen S C, Yun J X, Wang L H, He X J, Yu X M. Preparation of polyacrylamide-based supermacroporous monolithic cryogel beds under freezing-temperature variation conditions. Chemical Engineering Science, 2006, 61(20): 6701–6708

[28] Weng L H, Gouldstone A, Wu Y H, Chen W. Mechanically strong double network photo cross linked hydrogels from N,N-dimethylacrylamide and glycidyl methacrylated hyaluronan. Biomaterials, 2008, 29(14): 2153–2163

[29] Harley B A, Leung J H, Silva E C C M, Gibson L J. Mechanical characterization of collagen-glycosaminoglycan scaffolds. Acta Biomaterialia, 2007, 3(4): 463–474

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