Optimisation of propolis extracts production and in vitro study of protective qualities

Aim of the study: to determine optimal propolis extracts production conditions and study propolis protective qualities in vitro using HaCaT cell line. Research goals: 1. To execute propolis extracts production optimization. 2. To produce optimal propolis extracts and analyze their quality. 3. Adjust HaCaT cell line for the propolis protective qualities assessment. 4. Analyze optimal propolis extracts effect in vitro with HaCaT cell line. Research methods and the exploratory: propolis extracts production and optimization, identifying total phenolic compounds, while using spectrophotometry and identifying phenolic acids with chromatography, HaCaT cell line cultivation, propolis cytotoxicity and protective qualities analysis. Results: After spectrophotometry and chromatography analysis and in vitro study with HaCaT cell line, it was determined that using the smallest amount of propolis in the extraction process gave the best extraction of phenolic compounds (biggest amounts). Also it was determined that temperature influenced only the extracts that was made with water and 5% ethanol. According to the optimal propolis extracts and results, which was obtained after experiment, it clearly showed, that it did not recur. After analyzing data from the in vitro assay it was concluded that propolis is not cytotoxic and because of phenolic compounds it can protect cells from UV. Conclusions: 1. After analyzing the optimization process it was determined that the biggest influence for propolis extraction is the ratio between propolis quantity and solvent. Temperature influences extractions made with water and 5% ethanol. 2. When the optimal propolis extracts were made and the results compared to the ones that were proposed it was concluded that it does not recur. 3. Propolis extracts are able to protect HaCaT cell line from UV. The amount of cells in monolayer was bigger than control. 4. Optimal propolis extracts are not cytotoxic to HaCaT cell line. Recommendations: Propolis is a natural substance that has a lot of phenolic compounds. Its traditional use has already been examined by researchers all around the world. That is why it is recommended to continue studies for optimal propolis extracts production conditions. Also it is recommended to continue in vitro assays with HaCaT cell line, because there is not much data concerning this case. It is very important to continue this research, especially when the animal testing is continuously declined in preclinical trials