EVALUATION OF IN VITRO EFFECTS OF DIFFERENT AGENTS ON HYDATID CYST SCOLICES

Abstract

EVALUATION OF IN VITRO EFFECTS OF DIFFERENT AGENTS ON HYDATID CYST SCOLICESFurkan AYAN, Ertan BULBULOGLU, Semra OZCELİK, Mahfuz ELMASTAS, Ali TOPRAKObjective: Hydatid cyst is a common zoonotic disease that primarily affects the liver, typically caused by Echinococcus granulosus (1). Treatment options include follow-up, medical therapy, percutaneous procedures, and surgery (2). The recurrence rates vary among these treatments. To reduce recurrence rates, scolocidal agents are injected into the cyst during surgery and percutaneous intervention procedures. Hypertonic saline and ethanol are commonly used for injection, but there are also other agents with proven scolocidal activity (3). However, the agents used in intra-cystic injections can have unwanted side effects. The scolocidal agent used in intra-cystic injections should have high scolocidal activity, be effective in a short time, and have no undesirable side effects (4). Therefore, this study aims to contribute to the search for new scolocidal agents.Materials and Methods: During the operation of a patient with a liver cyst consistent with Gharbi type 3 hydatid cyst, cyst fluid and daughter cysts were collected. Daughter cysts were opened within the aspirated cyst fluid, and all the cyst contents were collected in glass cylinders. Protoscoleces were washed with phosphate-buffered saline. After a certain period, the protoscoleces settled, and the supernatant was discarded. Viability of protoscoleces was confirmed by their failure to be stained with eosin and their motility. In the examination 200,000 protoscoleces/cc were counted and the viability rate of over 95% was determined. Methanol extracts of Cinchona officinalis, Camellia sinensis, Commiphora molmol, and Artemisia annua were prepared in three different concentrations (25 mg/ml, 12.5 mg/ml, and 6.25 mg/ml) dissolved in dimethyl sulfoxide. Dimethyl sulfoxide and 20% hypertonic saline were used as control groups. 0,05 ml of each concentration of the agent to be tested was dispensed into wells of a well plate, and 0,05 ml of cyst fluid containing protoscoleces was added on top. The counts were performed by dropping 0,01% eosin solution onto slides together with each plant extract at 5, 15, and 25 minutes under a light microscope. Dead protoscoleces were stained red with eosin, while living protoscoleces were not stained. Each experiment was repeated three times. The number of dead scolices found under the light microscope was calculated as a percentage of the total scolices to determine the scolocidal activity.Results: Extracts of Camellia sinensis and Commiphora molmol did not exhibit effective scolocidal activity. Artemisia annua extract, at a concentration of 25 mg/ml, reached its highest effect at 25 minutes, with an average scolocidal activity of 87.25% at this time. The highest scolocidal activity was demonstrated by the Cinchona officinalis extract. When used at a concentration of 25 mg/ml, it was consistently more effective than 20% hypertonic saline across all time points and was the only agent to achieve 100% scolocidal activity at 25 minutes. The average scolocidal activity of 20% hypertonic saline was calculated to be 62.8% at 5 minutes, 76.95% at 15 minutes, and 89.65% at 25 minutes. Conclusion: This in vitro study demonstrates that Cinchona officinalis and Artemisia annua could serve as alternative scolocidal agents in the treatment of hydatid cysts.Keywords: Hydatid cyst, artemisia, cinchona References:1. Wen H, Vuitton L, Tuxun T, Li J, Vuitton DA, Zhang W, et al. Echinococcosis: Advances in the 21st Century. Clin Microbiol Rev. 2019 Mar 20;32(2). 2. Brunetti E, Kern P, Vuitton DA, Writing Panel for the WHO-IWGE. Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop. 2010 Apr;114(1):1–16. 3. Eckert J, International Office of Epizootics., World Health Organization. WHO/OIE manual on echinococcosis in humans and animals : a public health problem of global concern. World Organisation for Animal Health; 2001. 265 p. 4. Moazeni M, Nazer A. In vitro effectiveness of garlic (Allium sativum) extract on scolices of hydatid cyst. World J Surg. 2010 Nov;34(11):2677–81.