Hydrogels
that allow for the successful long-term in vitro culture
of cell–biomaterial systems to enable the maturation
of tissue engineering constructs are highly relevant in regenerative
medicine. Naturally derived polysaccharide-based hydrogels promise
to be one material group with enough versatility and chemical functionalization
capability to tackle the challenges associated with long-term cell
culture. We report a marine derived oxidized alginate, alginate dialdehyde
(ADA), and gelatin (GEL) system (ADA-GEL), which is cross-linked via ionic (Ca2+) and enzymatic (microbial transglutaminase,
mTG) interaction to form dually cross-linked hydrogels. The cross-linking
approach allowed us to tailor the stiffness of the hydrogels in a
wide range (from <5 to 120 kPa), without altering the initial ADA
and GEL hydrogel chemistry. It was possible to control the degradation
behavior of the hydrogels to be stable for up to 30 days of incubation.
Increasing concentrations of mTG cross-linker solutions allowed us
to tune the degradation behavior of the ADA-GEL hydrogels from fast
(<7 days) to moderate (14 days) and slow (>30 days) degradation
kinetics. The cytocompatibility of mTG cross-linked ADA-GEL was assessed
using NIH-3T3 fibroblasts and ATDC-5 mouse teratocarcinoma cells.
Both cell types showed highly increased cellular attachment on mTG
cross-linked ADA-GEL in comparison to Ca2+ cross-linked
hydrogels. In addition, ATDC-5 cells showed a higher proliferation
on mTG cross-linked ADA-GEL hydrogels in comparison to tissue culture
polystyrene control substrates. Further, the attachment of human umbilical
vein endothelial cells (HUVEC) on ADA-GEL (+) mTG was confirmed, proving
the suitability of mTG+Ca2+ cross-linked ADA-GEL for several
cell types. Summarizing, a promising platform to control the properties
of ADA-GEL hydrogels is presented, with the potential to be applied
in long-term cell culture investigations such as cartilage, bone,
and blood-vessel engineering, as well as for biofabrication.