Enhancement of aqueous solubility and antibiofilm activity of 4-allylpyrocatechol by polymeric micelles

Abstract

4-Allylpyrocatechol (APC), a major active compound of Piper betle, possesses strong antimicrobial activity. However, the water-insoluble property of APC limits its clinical and pharmaceutical use. To solve this problem, APC loaded polymeric micelles (PMAC) was fabricated using the thin-film hydration method. Nanoparticles of PMAC were characterized using a photon correlation spectrophotometer and transmission electron microscope (TEM). Antibiofilm activity of PMAC was investigated using crystal violet assay and confocal laser scanning microscopy (CLSM). Cytotoxic effects of PMAC on normal cells were investigated using MTT assay. The results demonstrate that a ratio of APC to the polymer plays an important role in the physicochemical characteristics of PMAC. The most suitable PMAC formulation having a small particle size (38.8 ± 1.4 nm), narrow size distribution (0.28 ± 0.10), a high negative zeta potential (− 16.43 ± 0.55 mV), and high entrapment efficiency (86.33 ± 14.27%) can be obtained from the ratio 1:4. The water solubility of this PMAC is significantly improved, approximately 1,000-fold higher than the unentrapped APC. TEM images demonstrate that PMAC is spherical in shape. The inhibitory effects of PMAC (1.5 mg APC/mL) against Streptococcus intermedius and Streptococcus mutans biofilms are significantly stronger than chlorhexidine (0.06 mg/mL). Images from CLSM demonstrate the destruction and thickness reduction of the pathogenic biofilms after contacting with PMAC. The MTT assay confirms that PMAC at this concentration is non-toxic to normal cells. These results obviously indicate that PMAC is a promising natural and harmless antimicrobial agent suitable for use in the oral cavity for inhibition of pathogenic bacterial biofilms.