The Fundamental Efficacy of Hesperidin-Loaded/chitosan-coated Hybrid Nanoparticles As a Prospective Regimen in Wound Healing Amendment: in Vitro and in Vivo Comprehensive Study

Hesperidin (Hsd), a promising bioactive phytomedicine, possesses a wound-healing activity. However, its poor solubility necessitates frequent topical application of high doses. Hence, this study aimed to prepare and characterize Hsd-loaded hybrid nanoparticles (Hsd-HNPs), followed by surface coating with chitosan (CS) to overcome such obstacles. Hsd was first encapsulated in a poly (D, L-lactic-co-glycolic) acid/soy phosphatidylcholine (PLGA/SPC) matrix, producing Hsd-HNPs, which were evaluated to choose the most appropriate one (F2). Then, F2 was coated with three concentrations of CS, and F2/CS1 was the optimal one, with a homogenous particle size of 254.10 +/- 4.59 nm, a positive zeta potential of 24.23 +/- 2.70 mV, a percent Hsd entrapping efficiency of 60.41 +/- 4.56 %, and a percent yield of 75.67 +/- 2.36 %. The stability of F2/CS1 was studied at a refrigerated temperature of 4 +/- 1 degrees C. Then, it was incorporated into a gel base and subjected to further investigation of ex vivo skin permeation and in vivo wound healing activity. F2/CS1 demonstrated good stability at 4 +/- 1 degrees C. The results of permeation studies manifested an enhanced pattern of Hsd permeation from F2/CS1 gel compared to that of the free Hsd. After topical application of F2/CS1 gel to the induced wound rat model, a significant and prominent wound healing efficacy was emphasized by histopathological and immunohistochemical examinations. In conclusion, Hsd/CS-HNPs (F2/CS1) gel could be regarded as promising topical nanophytomedicine with enhanced permeation and potentiated wound healing activity.