Engineering Three-dimensional Macroporous Hydroxyethyl Methacrylate-alginate-gelatin Cryogel for Growth and Proliferation of Lung Epithelial Cells

Three-dimensional (3D) growth of cell is of particular interest in the field of tissue engineering and regenerative medicine. Scaffolds used for this purpose are often tailor-made to mimic the microenvironment and the extracellular matrix of the tissue with defined role such as to provide appropriate structural, chemical, and mechanical support. The aim of the study was to design the macroporous matrix with potential in the field of tissue engineering especially for lung muscle regeneration. Blend of hydroxyethyl methacrylate-alginate-gelatin (HAG) cryogel scaffold was synthesized using cryogelation technique and this polymer material combination is being reported first time. The rheology study showed the elastic property of the material in wet state with no variation in storage modulus (G'), loss modulus (G″), and phase angle upon temperature variation. The microcomputer tomography (micro-CT) analysis confirmed the homogenous polymer structure with average pore diameter of 84 μm. Scaffold synthesized using polymer combinations which is mixture of polysaccharide (alginate) and protein (gelatin) provides supportive environment for human lung epithelial cell proliferation confirmed by cytoskeletal stain phalloidin and nuclei staining 4',6-diamidino-2-phenylindole checked for over three weeks. The in vivo biocompatibility was further performed which showed integration of scaffold to the surrounding tissue with ability to recruit cells. However, at first week, small amount of infiltrating mast cells were found which subsequently diminished in following weeks. Immunohistochemistry for dendritic cells confirmed in vivo biocompatible nature of the HAG scaffold. The mechanical strength, stiffness, elastic measurements, in vivo compatibility, and in vitro lung cell proliferation show the potentiality of HAG materials for lung tissue engineering.