In-situ assembly of Ca-alginate gels with controlled pore loading/release capability

Development of tailor-made porous polymer scaffolds acting as a temporary tissue-construct for cellular organization is of primary importance for tissue engineering applications. Control over the gel porosity is a critical issue due to the need for cells to proliferate and migrate and to ensure the transport of nutrition and metabolites. Gel loading with bioactive molecules is desired for target release of soluble signals to guide cell function. Calcium-alginate hydrogels are one of the most popular gels successfully utilized as polymer scaffolds. Here we propose a benchtop approach to design porous alginate gels by dispersion of CaCO3 vaterite crystals in sodium alginate followed by the crystal elimination. CaCO3 crystals play a triple role being (i) cross-linkers (a source of calcium ions to cross-link gel network), (ii) pore-makers (leaching of crystals retains the empty pores), and (iii) reservoirs with (bio)molecules (by molecule preloading into the crystals). Pore dimensions, interconnectivity, and density can be adjusted by choosing the size, concentration, and packing of the sacrificial CaCO3 crystals. An opportunity to load the pores with biomolecules was demonstrated using FITC-labeled dextrans of different molecular masses from 10 to 500 kDa. The dextrans were preloaded into CaCO3 vaterite crystals, and the subsequent crystal removal resulted in encapsulation of dextrans inside the pores of the gel. The dextran release rate from the gel pores depends on the equilibration of the gel structure as concluded by comparing dextran release kinetics during gelation (fast) and dextran diffusion into the performed gel (slower). Macromolecule binding to the gel is electrostatically driven as found for lysozyme and insulin. The application of porous gels as scaffolds potentially offering biomacromolecule encapsulation/release performance might be useful for alginate gel-based applications such as tissue engineering.