Page 105 - Book of Abstracts

5 International Scientific Online Conference DOI: https://doi.org/10.15414/2021.9788055224015

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EVALUATION OF THE DOSE-DEPENDENT EFFECTS OF TEA TREE OIL ON THE HEMOLYSIS
OF HUMAN ERYTHROCYTES
1
1
1
2
Maryna Opryshko , Myroslava Maryniuk , Oleksandr Gyrenko , Halyna Tkachenko ,
Natalia Kurhaluk ,Lyudmyla Buyun
1
2
1 M.M. Gryshko National Botanic Garden, National Academy of Science of Ukraine, Kyiv, Ukraine;
E-mail.: maryna.opryshko@meta.ua
2 Institute of Biology and Earth Sciences, Pomeranian University in Słupsk, Poland
Plant-derived alternative medicines such as tea tree (Melaleuca alternifolia (Maiden &
Betche) Cheel, Myrtaceae) oil have become increasingly popular in recent decades. Therefore,
the objectives of this study were to evaluate and compare the effects and three doses (final
concentrations were 20 and 10 μg/mL) of tea tree oil (TTO) on the hemolysis parameters of
human erythrocytes, as the step towards possible application as the anti-hemolytic agent.
The venous blood (10–20 ml) was obtained from normal volunteers via venipuncture. The
Research Ethics Committee of the Regional Medical Commission in Gdańsk (Poland) approved
the study (KB-31/18). The blood sample was incubated with TTA, respectively (final
concentrations were 20 and 10 μg/mL) at 25 °C for 15 min. The osmotic-induced hemolysis of
erythrocytes was measured spectrophotometrically with different 0.1–0.9 % NaCl solutions
(Mariańska et al., 2013). The assay is based on the measuring of the percent of erythrocytes
disintegration into hemolytic reagent action (0.1–0.9 % NaCl). The assay mixture contained 2.5
mL of NaCl solutions and 0.05 mL of 1 % erythrocyte suspension. The absorbance was read at
540 nm. The disintegration of erythrocytes (%) at different NaCl solutions was expressed as a
curve.
Treatment of the human erythrocytes by TTA in final concentration 20 μg/mL caused a
statistically significant increase of hemolysis in the samples with 0.9–0.5 % NaCl solutions: 20.2
% vs. 4.1 % (0.9 % NaCl), 26.4 % vs. 4.3 % (0.8 % NaCl), 27.5 % vs. 5.6 % (0.7 % NaCl), 32.9 %
vs. 5.7 % (0.6 % NaCl), and 36.8 % vs. 18.2 % (0.5 % NaCl). The increase of hemolysis was 4.9-
fold (0.9 % NaCl), 6.1-fold (0.8 % NaCl), 4.9-fold (0.7 % NaCl), 5.8-fold (0.6 % NaCl), 2-fold (0.5
% NaCl), respectively. In the sample with 0.4–0.1 % NaCl, a difference between the percentage
of hemolysis in samples after adding TTO to the erythrocytes compared to the untreated
samples was non-significant (83.7 %, 88.1 %, 91.9 %, and 94.6 % vs. 98.4 %, 98.5 %, 99 %, and
99.3 %). In the sample with 0.4–0.1 % NaCl, TTO in final concentration 20 μg/mL exhibited anti-
hemolytic activity after the treatment of human erythrocytes. TTO in final concentration 10
μg/mL caused a significant increase of hemolysis in the samples with 0.9–0.5 % NaCl solutions:
8.3 % vs. 1.23 % (0.9 % NaCl), 12.6 % vs. 1.23 % (0.8 % NaCl), 16.6 % vs. 1.5 % (0.7 % NaCl),
20.9 % vs. 1.97 % (0.6 % NaCl), and 24.2 % vs. 22.7 % (0.5 % NaCl). The increase of hemolysis
was 6.7-fold (0.9 % NaCl), 10.2-fold (0.8 % NaCl), 11.3-fold (0.7 % NaCl), 10.7-fold (0.6 % NaCl),
1.1-fold (0.5 % NaCl), respectively. After incubation in solutions of 0.4–0.1 % NaCl, adding of
TTO to the human erythrocytes caused the decrease of hemolysis (by 1.2 %, 4.3 %, 3.9 %, and
1.6 %, respectively).
Thus, based on the results from the hemolysis assay, it followed that TTO could affect the
human erythrocytes hemolysis in a dose-dependent manner. In particular, TTO seems to be
well tolerated by human erythrocytes down to a concentration of 5 μg/mL; higher quantities of
this TTO determined increasing impairment in a concentration-dependent manner as shown
by the results of the erythrogram models. This trend strengthens the necessity to test the toxic
effects of exogenous compounds directly on the erythrocytes when the aim is to use them in the
biological membrane model. The overall findings regarding the effects of the essential oil
derived from M. alternifolia are similar to the ones reported for human monocytes and
polymorphonuclear neutrophils.

Keywords: hemolysis, erythrocytes, tea tree essential oil, antioxidants.

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Spiritual Human Development
November 3 2021
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