In vitro anti-urolithiatic activity of hexane and methanol extracts of Celosia argentea and Biophytum sensitivum

Abstract

Kidney stone formation, also known as urolithiasis, is a worldwide health concern associated with lifestyle factors such as poor diet, dehydration, and obesity. The primary component of many stones is calcium oxalate. Although treatments like lithotripsy and pharmacotherapy are effective, they are expensive and invasive. Consequently, research has increasingly focused on plant-based treatments with anti-inflammatory and anti-crystallisation properties. In this study, the antiurolithiatic effects of hexane and methanol extracts from the root of Celosia argentea and the whole plant of Biophytum sensitivum were assessed in an in vitro setting. The respective concentration series 400, 600, 800, and 1000 μg/ml was prepared, and Potcit-XR®(positive control) and extract-free (negative control) were applied. The optical density (OD) of the tests (ODexperimental) and control samples (ODcotrol) was measured at 620 nm using a UV spectrophotometer following 30 minutes of incubation at 37◦C , performed in triplicate. The percentage inhibition of nucleation was calculated using the following formula: Percentage Inhibition = (1- ODexperimental) / (ODcotrol) × 100. IC50 was estimated by four-parameter logistic fitting where possible. Because the data had a non-parametric distribution, group differences were evaluated using the Kruskal-Wallis test, with pairwise comparisons conducted using Dunn’s test in GraphPad Prism. A p-value < 0.05 was considered statistically significant. All extracts inhibited nucleation in a dose-dependent manner. At 1000 μg/ml the % inhibition was: Positive control 59.5%, hexane C. argentea 56.5%, hexane B. sensitivum 38.1%, methanol B. sensitivum 38.6%, and methanol C. argentea 35.6%. Kruskal-Wallis indicated overall differences among groups (H = 22.92, p = 0.0001). Dunn’s post-hoc tests showed that the positive control significantly exceeded hexane B. sensitivum (p = 0.0006), methanol B. sensitivum (p = 0.0195) and methanol C. argentea (p < 0.0001). Hexane C. argentea was significantly more inhibitory than methanol C. argentea (p = 0.0049) and was not significantly different from the positive control (p = 0.14). The hexane C. argentea curve crossed 50% inhibition within the tested range; its IC50 was estimated at ∼500 μg/ml. Other extracts did not reach 50% by 1000 μg/ml (IC50) > 1000μg/mL). Hexane C. argentea demonstrated the strongest anti-nucleation activity among extracts and approached the efficacy of the reference drug, while B. sensitivum showed moderate activity and methanolic C. argentea was the weakest. The results imply that solvent polarity is a vital aspect of the extraction of bioactive components. Further research on standardised formulations, toxicity assessment, and enhancing effectiveness by combining the extracts with certain natural or synthetic compounds is suggested.