Evaluation of in vitro Anti-urolithiatic Activity of Hemidesmus indicus Sri Lankan Medicinal Plant used in Ayurvedic Medicine.

Abstract

Background: Urolithiasis is a prevalent condition affecting humans worldwide. The formation of calcium oxalate stones is the most common type, constituting about 80% of cases. Since currently available synthetic medications are associated with various side effects, there is a growing interest in complementary and/or alternative medicines due to their perceived safety, effectiveness, and lower cost. Objective: To evaluate the in vitro anti-urolithiatic activity of Hemidesmus indicus (Iramusu) in inhibiting the formation and growth of urinary stones Methods: Aqueous and hydro-alcoholic extracts of H. indicus was prepared by washing, drying, grinding, and using a Soxhlet apparatus. The aqueous extract used 15 g of plant material with 250 mL distilled water for 72 hours, while the hydro-alcoholic extract used 175 mL ethanol and 75 mL distilled water for 48 hours. Extracts were filtered, concentrated, freeze-dried, and stored at 4°C. Anti-urolithiatic activity was evaluated using nucleation and growth assays by preparing 1000 ppm to 62.5 ppm concentration series of stem, root, and leaf extracts to check the inhibition of calcium oxalate crystallization, by using cystone as positive control. Lethality of various extract concentrations was assessed by using in vitro brine shrimp assay. Results: When considering both nucleation and growth assays, growth assay showed the highest inhibition values. The resulting growth inhibition values of aqueous root extract were 44-65% and the 70% hydro-alcoholic extract exhibited 47-75%. For the aqueous stem extract, the inhibition values were from 19-25% and for the 70% hydro-alcoholic extract, it was 51-70%. For the aqueous leaf extract, the inhibition values were 26-59% and for the 70% hydro-alcoholic extract it was 14-36%. Out of the inhibition percentages, the aqueous extract of the root and 70% hydro alcoholic extract of the stem showed significant anti-urolithiatic activity (p<0.05). However, only the aqueous root and leaf extracts showed least toxic nature at the concentrations that showed the anti-urolithiatic activity. Conclusions: Comparative analysis indicates that aqueous extracts of root and 70% hydro-alcohol extract of stem exhibit significant anti-urolithiatic activity. Studies are recommended to formulate a solid dosage form by incorporating the aqueous root extract for the treatment of urolithiasis due to the least toxicity to overcome the limitations and adverse effects.