Composite Hydrogel of Poly(vinyl alcohol) Loaded by Citrus hystrix Leaf Extract, Chitosan, and Sodium Alginate with In Vitro Antibacterial and Release Test

Citrus hystrix leaves have been used traditionally as a spice, a traditional medicine for respiratory and digestive disorders, and a remedy for bacterial infections. This study reports on the synthesis of composite hydrogels using the freeze-thaw method with poly(vinyl alcohol) (PVA) as the building block loaded by C. hystrix leaf extract (CHLE). Additionally, chitosan (CS) and sodium alginate (SA) were also loaded, respectively, to increase the antibacterial activity and to control the extract release of the composite hydrogels. The combinations of the compositions were PVA, PVA/CHLE, PVA/CHLE/CS, PVA/CHLE/SA, and PVA/CHLE/SA/CS. The internal morphology of the hydrogels shows some changes after the PVA/CHLE hydrogel was loaded by CS, SA, and SA/CS. The analysis of the Fourier transform infrared (FTIR) spectra confirmed the presence of PVA, CHLE, CS, and SA in the composite hydrogels. From the X-ray diffraction (XRD) characterization, it was shown that the composite hydrogels maintained their semicrystalline properties with decreasing crystallinity degree after being loaded by CS, SA, and SA/CS, as also supported by differential scanning calorimetry (DSC) characterization. The compressive strength of the PVA/CHLE hydrogel decreases after the loading of CS, SA, and SA/CS, so that it becomes more elastic. Despite being loaded in the composite hydrogels, the CHLE retained its antibacterial activity, as evidenced in the in vitro antibacterial test. The loading of CS succeeded in increasing the antibacterial activity of the composite hydrogels, while the loading of SA resulted in the decrease of the antibacterial activity. The release of extract from the composite hydrogels was successfully slowed down after the loading of CS, SA, and SA/CS, resulting in a controlled release following the pseudo-Fickian diffusion. The cytotoxic activity test proved that all hydrogel samples can be used safely on normal cells up to concentrations above 1000 mu g/mL.