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Jove-Journal of Visualized Experiments

Jove-Journal of Visualized Experiments

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Synthesis of Thermogelling Poly(N-isopropylacrylamide)-graft-chondroitin Sulfate Composites with Alginate Microparticles for Tissue Engineering.

Published:26 October 2016 DOI: 10.3791/53704
T. Christiani,?K. Toomer,?J. Sheehan,?A. Nitzl,?A. Branda,?E. England,?P. Graney,?C. Iftode,?A. Vernengo

Abstract

Injectable biomaterials are defined as implantable materials that can be introduced into the body as a liquid and solidify in situ. Such materials offer the clinical advantages of being implanted minimally invasively and easily forming space-filling solids in irregularly shaped defects. Injectable biomaterials have been widely investigated as scaffolds for tissue engineering. However, for the repair of certain load-bearing areas in the body, such as the intervertebral disc, scaffolds should possess adhesive properties. This will minimize the risk of dislocation during motion and ensure intimate contact with the surrounding tissue, providing adequate transmission of forces. Here, we describe the preparation and characterization of a scaffold composed of thermally sensitive poly(N-isopropylacrylamide)-graft-chondroitin sulfate (PNIPAAM-g-CS) and alginate microparticles. The PNIPAAm-g-CS copolymer forms a viscous solution in water at RT, into which alginate particles are suspended to enhance adhesion. Above the lower critical solution temperature (LCST), around 30 °C, the copolymer forms a solid gel around the microparticles. We have adapted standard biomaterials characterization procedures to take into account the reversible phase transition of PNIPAAm-g-CS. Results indicate that the incorporation of 50 or 75 mg/ml?alginate particles into 5% (w/v) PNIPAAm-g-CS solutions quadruple the adhesive tensile strength of PNIPAAm-gCS alone (p<0.05). The incorporation of alginate microparticles also significantly increases swelling capacity of PNIPAAm-g-CS (p<0.05), helping to maintain a space-filling gel within tissue defects. Finally, results of the in vitro toxicology assay kit, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and Live/Dead viability assay indicate that the adhesive is capable of supporting the survival and proliferation of encapsulated Human Embryonic Kidney (HEK) 293 cells over 5 days.

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Related products
Procduct Name CAS Molecular Formula Supplier Price
Sodium alginate 9005-38-3 C5H7O4COONa 794 suppliers $12.70-$3270.00
XTT sodium salt 111072-31-2 C22H18N7NaO13S2 228 suppliers $45.00-$1190.00
POLY(N-ISOPROPYL ACRYLAMIDE) 25189-55-3 C6H11NO 96 suppliers $29.00-$902.00
Human embryonic kidney cells 293 C18 9 suppliers Inquiry
Alginate Antibody 8 suppliers Inquiry
XTT (2,3-BIS-(2-METHOXY-4 5 suppliers Inquiry

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