ACUTE TOXICITY OF PAPAYA (CARICA PAPAYA L. VAR CALINA) FRUIT AND SEED EXTRACTS AND THEIR POTENTIAL ANTI-INFLAMMATORY ACTIVITY IN CAUDAL FIN AMPUTATION-INDUCED ZEBRAFISH LARVAE (DANIO RERIO)

• Main Author: Kharisma Primastuti, Rahmi
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/76061
• Institution: Institut Teknologi Bandung

<p align="justify">Inflammation is the body's immune response to various stimuli, such as pathogens or toxic compounds, and can be harmful to the body if it becomes excessive. Drugs in the steroid and non-steroidal classes can help relieve the symptoms of inflammation, such as redness, swelling, heat, and pain. However, the use of these drugs can cause reactions that are harmful to the body. Several studies have investigated the potential of plant extracts, especially papaya (Carica papaya L. var Calina), as therapeutic agents. However, no study has directly compared the anti-inflammatory potential of papaya fruit and seed extracts. Therefore, the effect of papaya fruit and seed extracts on the innate immune response and wound tissue regeneration process in experimental animals was investigated in this study. Toxicity tests of the extracts were first conducted to determine the toxic effects and risks that may be caused by the administration of the extracts to experimental animals. The extracts were exposed by the immersion method. Zebrafish (Danio rerio) embryos at 10 hpf/hour post fertilization (n=60/group) were maintained in medium E3 1X + 0.003% PTU for 24 hours. Embryos at 1 dpf/day post fertilization were exposed to each test and control solution. The test solutions consisted of water-solvent papaya fruit extract (BUPA), methanol-solvent papaya fruit extract (BUPM), water-solvent papaya seed extract (BIPA), and methanol-solvent papaya seed extract (BIPM), each at LC25 concentration. The negative control was medium E3 1X + 0.003% PTU, while the positive control was dexamethasone (DEX). After 48 hours of rearing, caudal fin amputation was performed on zebrafish larvae (3 dpf) and then stained. Neutral red stain was used to identify macrophages, while Sudan black stain was used to identify neutrophils. The staining was performed at 4 and 48 hours after amputation of the caudal fin. The toxicity tests showed that the extracts of the papaya fruit and seeds are in toxicity category 4. That means they are harmless. The results showed that treatment with each extract had a significant effect on reducing the number of macrophages and neutrophils at the injury site compared to the negative control at 4 hours of observation (p<0.05). However, at 48 hours, some of the treatment results became insignificant compared to the negative control (p>0.05). All extract treatments were not significantly different from the positive control (p>0.05). All extract treatments had a significant effect on reducing the regeneration area compared to the negative and positive control (p<0.05). This shows that papaya seed and fruit extracts can inhibit the migration of neutrophils and macrophages to the site of injury, resulting in the inhibition of the regeneration process of the caudal fin of zebrafish larvae. This result was expected because based on the GC-MS results, fatty acid groups, hydrocarbons, and aromatic compounds were found to have anti-inflammatory and antioxidant activities. Papaya seed extract consistently showed lower macrophage and neutrophil counts than papaya fruit extract. This suggests that papaya seed extract may have greater potential to reduce the number of macrophages and neutrophils at the site of injury compared to papaya fruit extract.