experiment.
One way to apply this type of hydrogels for drug delivery and tissue engineering is to use them as wound dressing (Hamedi, 2018; Wechsler, 2019). Thereby, the hydrogel absorbs wound exudates, providing a balance of hydration and, also, using this aqueous environment to slowly release loaded drugs. In order to evaluate the general behavior of starch-CS-GA hydrogel in vivo , it was used in a mouse wound healing model (Figure 9 ). No nanoparticles were used in this experiment, only hydrogel effect was evaluated.
All wounds in control and treated animals had a 6 mm diameter on day 0; variations of this parameter were studied for 15 days (Figure 9A ). At the 3rd day of the experiment, mice treated with hydrogels presented a dry wound, without redness or inflammation. In contrast, control animals showed inflamed, reddened, and exuding wounds. Because of this, an increase in the diameter of the wounds was observed in untreated animals, presenting a deterioration instead of the recovery observed in hydrogel-treated animals (Figure 9B ). A similar effect was observed on day 6 for both groups. But, around a week after the beginning of the experiment, the natural healing process was evident in control mice, which showed a reduction in wound diameters at day 9. However, wound healing effects were more evident in the treated animals. By day 12, all treated animals healed, while this process was not completed in untreated control mice.
The study revealed significative differences between both groups in wound healing during the complete experiment. The absence of inflammation and exudates in treated animals, along with faster healing, suggests the hydrogel beneficial effect. These results evidence the potentialities of starch-CS-GA hydrogel for wound dressing applications. Although no biologically active drugs or nanoparticles were used in thisin vivo study, the swelling properties and the efficient load of chitosan nanoparticles observed for this type of hydrogel, are encouraging evidences of its potential application in drug delivery.
Conclusions .
Based on its excellent properties, such as biodegradability, biocompatibility, and low cost, starch is a remarkably interesting choice for the development of polymer-based devices in biological applications. Its combination with other polymers could improve the mechanical and biological properties and facilitate the formation of functional hydrogels. PVA and chitosan interaction with starch led to the formation of hydrogels under certain conditions. Among the four synthesized hydrogels, the combination of starch and chitosan by chemical crosslinking showed the best results in terms of structure, swelling, and resulted safe in vitro and in vivo . The wound healing experiment in mice proved the potentialities of starch-CS-GA for tissue engineering applications; while its excellent interaction with chitosan nanoparticles evidenced a promissory behavior in the delivery of these nanoparticulate systems. The other three hydrogels presented relevant properties that justify next studies in order to fully exploit their potential.
Conflict of interest .
The authors have no conflict of interest to declare.
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Table .