Abstract
A covalently cross-linked injectable hybrid hydrogel, namely, alginate/poly(amidoamine) (PAMAM), with the objective of cell delivery was innovatively designed and synthesized using tetra-amino-functional PAMAM dendrimer as the cross-linker. With the increase in percentage of PAMAM cross-linker, the pore size and swelling ratio of hydrogels were in the range of 57 ± 18 μm to 88 ± 25 μm and 110 ± 16 to 157 ± 20, respectively. The study of attachment and proliferation of MC3T3-E1 pre-osteoblasts using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay through indirect and direct contact methods indicated a continuous increase in metabolically active live cells with time, implying non-cytotoxicity of the synthesized hydrogel. The live-dead assay showed >95% of live cells for alginate/PAMAM hydrogels, suggesting viability of the encapsulated cells. When the percentage of PAMAM cross-linker in alginate/PAMAM hydrogel was increased from 5 to 25, the percentage degradation rate decreased from 1.1 to 0.29%/day. Given that the poly(ethylene glycol) is commonly used cross-linker for hydrogel syntheses, we compared the behavior with poly(ethylene glycol). The incorporation of poly(ethylene glycol) in alginate/PAMAM hydrogel reduced the activity of MC3T3-E1 cells and their viability compared to the alginate/PAMAM hydrogels. The protonation of amino groups in alginate/PAMAM injectables under physiological conditions led to the formation of cationic hydrogels. These cationic hydrogels showed enhanced cell encapsulation and attachment ability because of electrostatic interaction with negatively charged cell surface as determined by cell adhesion and extensions from scanning electron microscope and vinculin assay and ability of in situ calcium phosphate mineralization. These observations point toward the potential use as an injectable scaffold for cell delivery and tissue engineering applications.
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