Preparation and In Vivo Toxicity Study of Allantoin Incorporated Hyaluronic Acid-L-Cysteine Oral Solution: A Future Treatment for Mucositis

Background: Thiolated hyaluronic acid (HA) with interesting properties, such as mucoadhesiveness, enzyme inhibitory, permeation enhancing, and release controlling properties can be applied for drug delivery in various diseases like mucositis. The purpose of this study was to evaluate the stability and toxicity of thiol modified HA by the aid of L-cysteine ethyl ester hydrochloride (Cys) named (HA-Cys) and allantoin (Alla) incorporated HA-Cys (HA-Cys-Alla) to reveal their potential for the future treatment of mucositis. Methods: The HA modification and drug incorporation were investigated using FTIR spectroscopy. The evaluation of in vitro cytotoxicity on Caco-2 cell line by means of MTT assay and in vivo toxicity by measuring the hematological and biochemical parameters in rats were performed. The appearance stability of HA-Cys and HA-Cys-Alla was evaluated at room and refrigerator temperatures over time. In addition, the stability of HA-Cys and HA-Cys-Alla subjected to heating and cooling, freeze-thaw, centrifugal forces, as well as the pH stability under the above-mentioned conditions were also investigated. Results: The results indicated that the synthesized HA-Cys and HA-Cys-Alla with pseudo-plastic rheological behavior demonstrated excellent stability at refrigerator temperature. Although HA-Cys showed good stability, the HA-Cys-Alla revealed color change at room temperature. Moreover, despite no much resistance of HA-Cys and HA-Cys-Alla against the heating-cooling test, the samples exhibited good resistance against freeze-thaw and centrifugal forces. Also, convenient pH stability and high in vitro and in vivo biocompatibility were observed. Conclusion: The low in vitro and in vivo toxicity and convenient stability of HA-Cys-Alla has introduced it as a proper candidate for future clinical applications.