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Hydrogels show great potential particularly in the development of drug delivery because of their desirable properties. The enzymatic crosslinked casein hydrogel is a new generation and has received great attention for drug delivery. In the present study, the enzymatic crosslinked casein hydrogel containing insulin was prepared and in vitro and in vivo evaluations were assessed. The hydrogels were also evaluated using differential scanning calorimeter; X-ray diffraction; Scanning electron microscopy and Atomic force microscopy methods. The rheological and releasing behaviors at different media were also investigated. The secondary structure of insulin was assayed by CD method and finally, the hypoglycemic effects were tested in the diabetic rats. Hydrogel showed a three-dimensional porous structure. Rheographs indicated the combination of thixotropic and pseudoplastic behaviors. In vitro release studies confirmed that insulin released in acidic condition slowly, however, in neutral and alkaline environments insulin rapidly released. The structure of insulin preserved following release. Hypoglycemic effects were observed by oral administration of insulin-loaded hydrogel. Thus, the enzymatic crosslinked hydrogel can be considered as a good candidate for oral delivery of insulin.

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