Formulation and evaluation of encapsulated Graptophyllum pictum (L.) Griff. ethanolic extract in lipid vesicles for hemorrhoid treatment

Context: Graptophyllum pictum (L.) Griff. leaves (GPL) are traditionally used in Indonesia to treat hemorrhoids, and the anti-hemorrhoid activity of the ethanolic extract of G. pictum leaves (GPLE) has been proven scientifically. However, since it contains a variety of compounds, both hydrophilic and lipophilic, which has diverse solubility, absorption, and penetration characteristic, there is a need to be formulated to improve its effectiveness. Lipid nanocarrier formulations, such as liposome and ethosome, can be developed as the delivery system for GPLE to enhance its effectiveness. Aims: To evaluate the anti-hemorrhoid activity of the liposomal and ethosomal formulation of GPLE. Methods: The hydration method was used to prepare GPLE liposomal (GL) and ethosomal (GE) formulations. An in vivo study was conducted to evaluate the penetration and effectiveness of both formulations. The in vivo test was evaluated based on the recto-anal tissue histological observation after treatment of the formulation to croton oil-induced hemorrhoid mice. The number of inflammatory cells, goblet cells, hemorrhage area, and mucosal thickness were compared between both formulations and the extract. Results: GE had a smaller particle size, 145.7 ± 5.75 nm, with a polydispersity index (PDI) value of 0.376 ± 0.031. Meanwhile, GL showed a particle size of 407.80 ± 11.32 nm, with a PDI value of 0.389 ± 0.019. The zeta potential (ZP) of GL was -2.0200 ± 0.0513 mV and -0.0109 ± 0.0257 mV for GE. GE exhibited a better penetration and activity profile than GL, which was assumed to be correlated to its smaller particle size and component of the formulation. GE was able to penetrate up to the submucosa layer; meanwhile, GL only reached the muscularis mucosa. GE revealed better anti-hemorrhoid activity as it showed a lesser number of inflammatory cells, goblet cells, and necrosis cells. Additionally, it exhibited a smaller hemorrhage area and muscular mucose cell thickness compared to GL. However, there was no significant difference statistically. Conclusions: These findings suggest that lipid vesicles are a suitable delivery system for GPLE in the anti-hemorrhoid treatment.