Research Article
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The analysis of the effects of propolis products as food supplements on the viability of baby hamster kidney (BHK-21) and murine macrophage (RAW 264.7) cells by spectrophotometric MTT assay

Year 2022, Volume: 7 Issue: 1, 45 - 53, 30.04.2022
https://doi.org/10.31797/vetbio.970578

Abstract

Propolis is beeswax with rich bioactive compound content. On the market, there many propolis products as food supplements for the consumers. This study aimed to investigate the effect of food supplement products based on propolis at several concentrations on the viability of baby hamster kidney cells and murine macrophage cell lines (BHK-21 and RAW 264.7). For this purpose, both cell lines were treated with the two-fold serial dilutions (from 20 to 2-10) of each six propolis products (P1, P2, P3, P4, P5 and P6) after reaching monolayer cell in 96-well microplates. The viability and inhibition of cells were spectrophotometrically determined by MTT assay after 24 h. For BHK-21, the IC50s of P1, P2, P3, P4, P5 and P6 were calculated as 0.003, 0.178, 0,082, 0.451, 0.278 and 0.384 %, respectively. For RAW 264.7, the IC50 of P1, P2, P3, P4, P5 and P6 were calculated as 0.260, 0.218, 0.115, 0.257, 0.207 and 0.265%, respectively. The IC50 value was higher for RAW 264.7 cells than for BHK-21 cells. So, the low cytotoxic effect was determined in RAW 264.7 cells. Propolis products contain some additives (aroma, flavouring) had lower the IC50 and the lower viability of BHK-21 cells. So, Additives in the propolis food supplement might be an effective factor on cell viability as much as dilution factor and propolis content.

Supporting Institution

Yok

Project Number

Yok

Thanks

The authors thank the Institute of Health Sciences and the Faculty of Health Sciences at Marmara University.

References

  • Valcic, S., Montenegro, G., Mujica, A., Avila, G., Franzblau, S., Singh, M., Maiese, W. & Timmermann, B. (1999). Phytochemical, Morphological, and Biological Investigations of Propolis from Central Chile. Zeitschrift für Naturforschung C, 54(5-6), 406-416. https://doi.org/10.1515/znc-1999-5-617
  • Gojmerac W. (1980). Bees, beekeeping, honey and pollination. Westport, USA: A VI publishing Inc, Burdock G. A. (1998). Review of the biological properties and toxicity of bee propolis (propolis). Food and Chemical Toxicology, 36(4), 347–363. https://doi.org/10.1016/s0278-6915(97)00145-2
  • Yaghoubi, M., & Ghorbani, G., & Soleymanianzad, S., & Sattari, R. (2007). Antimicrobial activity of iranian propolis and its chemical composition. Daru Journal of Pharmaceutical Science, 15(1), 45-48.
  • Markham, K.R., Mitchell, K., Wilkins, A., Daldy, J.A., & Lu, Y. (1996). HPLC and GC-MS identification of the major organic constituents in New Zealand propolis. Phytochemistry, 42, 205-211. https://doi.org/10.1016/0031-9422(96)83286-9
  • Sforcin, J. M., Fernandes, A., Jr, Lopes, C. A., Bankova, V., & Funari, S. R. (2000). Seasonal effect on Brazilian propolis antibacterial activity. Journal of Ethnopharmacology, 73(1-2), 243–249. https://doi.org/10.1016/s0378-8741(00)00320-2
  • Al-Shaher, A., Wallace, J., Agarwal, S., Bretz, W., & Baugh, D. (2004). Effect of propolis on human fibroblasts from the pulp and periodontal ligament. Journal of Endodontics, 30(5), 359–361. https://doi.org/10.1097/00004770-200405000-00012
  • Bedier, M. M. (2016). Effect of mineral trioxide aggregate with or without propolis extract on the proliferation of fibroblast cell line. Tanta Dental Journal, 13(4), 208.
  • Martini, A.A.K., & Mahendra, A. N. (2019). Administration of 50% propolis ethanolic extract increases the number of gingivitis fibroblast in H2O2 -induced rats. AIP Conference Proceedings 2019. https://doi.org/10.1063/1.5110021
  • Bogdanov, S. (2014). Propolis: Composition, Health, Medicine: A Review. Bee Product Science, 1-40.
  • Berretta, A., Silveira, M.D., Capcha, J.M., & Jong, D.D. (2020). Propolis and its potential against SARS-CoV-2 infection mechanisms and COVID-19 disease. Biomedicine & Pharmacotherapy, 131, 110622 - 110622. https://doi.org/10.1016/j.biopha.2020.110622
  • Moussa, M., & Temirek, M. (2018). Biological effect of Egyptian Propolis on BHK cell line and on dental pulp tissue (An In vitro And In vivo study). Egyptian Dental Journal, 64, 2161-2170. https://doi.org/10.21608/edj.2018.76780
  • Cox, C. F., Bergenholtz, G., Heys, D. R., Syed, S. A., Fitzgerald, M., & Heys, R. J. (1985). Pulp capping of dental pulp mechanically exposed to oral microflora: a 1-2 year observation of wound healing in the monkey. Journal of oral pathology, 14(2), 156–168. https://doi.org/10.1111/j.1600-0714.1985.tb00479.x
  • Kurniati, S. I. (2018). Effect of 10% propolis microgel application to odontoblast cell line viability in external bleaching, [Master Thesis, Hasanuddin Makassar University]
  • Wardati, Y., Mooduto, L., & Prasetyo, E. A. (2014). Biocompatibility evaluation of Propolis extract and H2O2 3% to Baby Hamster Kidney-21 (BHK-21) fibroblast. Conservative Dentistry Journal, 4 (1), 34-39.
  • Kartika, W.P., Kunarti, S., & Subiyanto, A. (2015). Genotoxicity test of propolis extract, mineral trioksida aggregat, and calcium hydroxide on fibroblast BHK-21 cell cultures. Dental Journal: Majalah Kedokteran Gigi, 48, 16-21. http://dx.doi.org/10.20473/j.djmkg.v48.i1.p16-21
  • Widjiastuti, I., Dewi, M., Prasetyo, E., Pribadi, N., & Moedjiono, M. (2020). The cytotoxicity test of calcium hydroxide, propolis, and calcium hydroxide-propolis combination in human pulp fibroblast. Journal of Advanced Pharmaceutical Technology & Research, 11, 20 - 24. https://doi.org/10.20473/J.DJMKG.V48.I1.P16-21
  • Aliyazicioglu, Y., Demir, S., Turan, I., Cakiroglu, T. N., Akalin, I., Deger, O., & Bedir, A. (2011). Preventive and protective effects of Turkish propolis on H₂O₂-induced DNA damage in foreskin fibroblast cell lines. Acta biologica Hungarica, 62(4), 388–396. https://doi.org/10.1556/ABiol.62.2011.4.5
  • Al-Haj Ali, S. N. (2016). In vitro toxicity of propolis in comparison with other primary teeth pulpotomy agents on human fibroblasts. Journal of investigative and clinical dentistry, 7(3), 308–313. https://doi.org/10.1111/jicd.12157
  • Murase, H., Shimazawa, M., Kakino, M., Ichihara, K., Tsuruma, K., & Hara, H. (2013). The effects of brazilian green propolis against excessive light-induced cell damage in retina and fibroblast cells. Evidence-Based Complementary and Alternative Medicine, 238279. https://doi.org/10.1155/2013/238279
  • Misir, S., Aliyazicioglu, Y., Demir, S., Turan, I., Yaman, S., & Deḡer, O. (2018). Antioxidant properties and protective effect of turkish propolis on t-bhp-induced oxidative stress in foreskin fibroblast cells. Indian Journal of Pharmaceutical Education and Research, 52, 94-100. https://doi.org/10.5530/ijper.52.1.11
  • Mooduto, L., Fredline, C., Sosiawan, A., & Wahjuningrum, D. (2019). Cytotoxicity of sodium hypochlorite, chlorhexidine and propolis on human periodontal ligament fibroblast cell. Journal of International Dental and Medical Research, 12(2), 476-480.
  • Uğur Aydin, Z., Akpinar, K. E., Hepokur, C., & Erdönmez, D. (2018). Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells. Brazilian Oral Research, 32, 119. https://doi.org/10.1590/1807-3107bor-2018.vol32.0119
  • Sahlan, M., Mahira, K. F., Pratami, D. K., Rizal, R., Ansari, M. J., Al-Anazi, K. M., & Farah, M. A. (2021). The cytotoxic and anti-inflammatory potential of Tetragonula sapiens propolis from Sulawesi on raw 264.7 cell lines. Journal of King Saud University - Science, 33(2), 101314. https://doi.org/10.1016/j.jksus.2020.101314
  • Han, S., Sung, K. H., Yim, D., Lee, S., Cho, K., Lee, C. K., Ha, N. J., & Kim, K. (2002). Activation of murine macrophage cell line RAW 264.7 by Korean propolis. Archives of Pharmacal Research, 25(6), 895-902. https://doi.org/10.1007/BF02977011
  • Kim, S. H., Kim, M. O., Kim, E. K., & Kim, K. R. (2019). Antiinflammatory effects of korean propolis in lipopolysaccharide-stimulated RAW 264.7 macrophages and pma-induced mouse ear edema. Indian Journal of Pharmaceutical Sciences, 81(3), 533-538. https://doi.org/10.36468/pharmaceutical-sciences.540
  • Asgharpour, F., Moghadamnia, A. A., Motallebnejad, M., & Nouri, H. R. (2019). Propolis attenuates lipopolysaccharide-induced inflammatory responses through intracellular ROS and NO levels along with downregulation of IL-1β and IL-6 expressions in murine RAW 264.7 macrophages. Journal of food biochemistry, 43(8), e12926. https://doi.org/10.1111/jfbc.12926
  • Myint, T. M., Tezuka, Y., Midorikawa, K., & Matsushige, K. (2003). Inhibitors of nitric oxide (NO) production in murine macrophage-like J774. 1 cells from Brazilian propolis. Journal of traditional medicines, 20(1), 22-29.
Year 2022, Volume: 7 Issue: 1, 45 - 53, 30.04.2022
https://doi.org/10.31797/vetbio.970578

Abstract

Project Number

Yok

References

  • Valcic, S., Montenegro, G., Mujica, A., Avila, G., Franzblau, S., Singh, M., Maiese, W. & Timmermann, B. (1999). Phytochemical, Morphological, and Biological Investigations of Propolis from Central Chile. Zeitschrift für Naturforschung C, 54(5-6), 406-416. https://doi.org/10.1515/znc-1999-5-617
  • Gojmerac W. (1980). Bees, beekeeping, honey and pollination. Westport, USA: A VI publishing Inc, Burdock G. A. (1998). Review of the biological properties and toxicity of bee propolis (propolis). Food and Chemical Toxicology, 36(4), 347–363. https://doi.org/10.1016/s0278-6915(97)00145-2
  • Yaghoubi, M., & Ghorbani, G., & Soleymanianzad, S., & Sattari, R. (2007). Antimicrobial activity of iranian propolis and its chemical composition. Daru Journal of Pharmaceutical Science, 15(1), 45-48.
  • Markham, K.R., Mitchell, K., Wilkins, A., Daldy, J.A., & Lu, Y. (1996). HPLC and GC-MS identification of the major organic constituents in New Zealand propolis. Phytochemistry, 42, 205-211. https://doi.org/10.1016/0031-9422(96)83286-9
  • Sforcin, J. M., Fernandes, A., Jr, Lopes, C. A., Bankova, V., & Funari, S. R. (2000). Seasonal effect on Brazilian propolis antibacterial activity. Journal of Ethnopharmacology, 73(1-2), 243–249. https://doi.org/10.1016/s0378-8741(00)00320-2
  • Al-Shaher, A., Wallace, J., Agarwal, S., Bretz, W., & Baugh, D. (2004). Effect of propolis on human fibroblasts from the pulp and periodontal ligament. Journal of Endodontics, 30(5), 359–361. https://doi.org/10.1097/00004770-200405000-00012
  • Bedier, M. M. (2016). Effect of mineral trioxide aggregate with or without propolis extract on the proliferation of fibroblast cell line. Tanta Dental Journal, 13(4), 208.
  • Martini, A.A.K., & Mahendra, A. N. (2019). Administration of 50% propolis ethanolic extract increases the number of gingivitis fibroblast in H2O2 -induced rats. AIP Conference Proceedings 2019. https://doi.org/10.1063/1.5110021
  • Bogdanov, S. (2014). Propolis: Composition, Health, Medicine: A Review. Bee Product Science, 1-40.
  • Berretta, A., Silveira, M.D., Capcha, J.M., & Jong, D.D. (2020). Propolis and its potential against SARS-CoV-2 infection mechanisms and COVID-19 disease. Biomedicine & Pharmacotherapy, 131, 110622 - 110622. https://doi.org/10.1016/j.biopha.2020.110622
  • Moussa, M., & Temirek, M. (2018). Biological effect of Egyptian Propolis on BHK cell line and on dental pulp tissue (An In vitro And In vivo study). Egyptian Dental Journal, 64, 2161-2170. https://doi.org/10.21608/edj.2018.76780
  • Cox, C. F., Bergenholtz, G., Heys, D. R., Syed, S. A., Fitzgerald, M., & Heys, R. J. (1985). Pulp capping of dental pulp mechanically exposed to oral microflora: a 1-2 year observation of wound healing in the monkey. Journal of oral pathology, 14(2), 156–168. https://doi.org/10.1111/j.1600-0714.1985.tb00479.x
  • Kurniati, S. I. (2018). Effect of 10% propolis microgel application to odontoblast cell line viability in external bleaching, [Master Thesis, Hasanuddin Makassar University]
  • Wardati, Y., Mooduto, L., & Prasetyo, E. A. (2014). Biocompatibility evaluation of Propolis extract and H2O2 3% to Baby Hamster Kidney-21 (BHK-21) fibroblast. Conservative Dentistry Journal, 4 (1), 34-39.
  • Kartika, W.P., Kunarti, S., & Subiyanto, A. (2015). Genotoxicity test of propolis extract, mineral trioksida aggregat, and calcium hydroxide on fibroblast BHK-21 cell cultures. Dental Journal: Majalah Kedokteran Gigi, 48, 16-21. http://dx.doi.org/10.20473/j.djmkg.v48.i1.p16-21
  • Widjiastuti, I., Dewi, M., Prasetyo, E., Pribadi, N., & Moedjiono, M. (2020). The cytotoxicity test of calcium hydroxide, propolis, and calcium hydroxide-propolis combination in human pulp fibroblast. Journal of Advanced Pharmaceutical Technology & Research, 11, 20 - 24. https://doi.org/10.20473/J.DJMKG.V48.I1.P16-21
  • Aliyazicioglu, Y., Demir, S., Turan, I., Cakiroglu, T. N., Akalin, I., Deger, O., & Bedir, A. (2011). Preventive and protective effects of Turkish propolis on H₂O₂-induced DNA damage in foreskin fibroblast cell lines. Acta biologica Hungarica, 62(4), 388–396. https://doi.org/10.1556/ABiol.62.2011.4.5
  • Al-Haj Ali, S. N. (2016). In vitro toxicity of propolis in comparison with other primary teeth pulpotomy agents on human fibroblasts. Journal of investigative and clinical dentistry, 7(3), 308–313. https://doi.org/10.1111/jicd.12157
  • Murase, H., Shimazawa, M., Kakino, M., Ichihara, K., Tsuruma, K., & Hara, H. (2013). The effects of brazilian green propolis against excessive light-induced cell damage in retina and fibroblast cells. Evidence-Based Complementary and Alternative Medicine, 238279. https://doi.org/10.1155/2013/238279
  • Misir, S., Aliyazicioglu, Y., Demir, S., Turan, I., Yaman, S., & Deḡer, O. (2018). Antioxidant properties and protective effect of turkish propolis on t-bhp-induced oxidative stress in foreskin fibroblast cells. Indian Journal of Pharmaceutical Education and Research, 52, 94-100. https://doi.org/10.5530/ijper.52.1.11
  • Mooduto, L., Fredline, C., Sosiawan, A., & Wahjuningrum, D. (2019). Cytotoxicity of sodium hypochlorite, chlorhexidine and propolis on human periodontal ligament fibroblast cell. Journal of International Dental and Medical Research, 12(2), 476-480.
  • Uğur Aydin, Z., Akpinar, K. E., Hepokur, C., & Erdönmez, D. (2018). Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells. Brazilian Oral Research, 32, 119. https://doi.org/10.1590/1807-3107bor-2018.vol32.0119
  • Sahlan, M., Mahira, K. F., Pratami, D. K., Rizal, R., Ansari, M. J., Al-Anazi, K. M., & Farah, M. A. (2021). The cytotoxic and anti-inflammatory potential of Tetragonula sapiens propolis from Sulawesi on raw 264.7 cell lines. Journal of King Saud University - Science, 33(2), 101314. https://doi.org/10.1016/j.jksus.2020.101314
  • Han, S., Sung, K. H., Yim, D., Lee, S., Cho, K., Lee, C. K., Ha, N. J., & Kim, K. (2002). Activation of murine macrophage cell line RAW 264.7 by Korean propolis. Archives of Pharmacal Research, 25(6), 895-902. https://doi.org/10.1007/BF02977011
  • Kim, S. H., Kim, M. O., Kim, E. K., & Kim, K. R. (2019). Antiinflammatory effects of korean propolis in lipopolysaccharide-stimulated RAW 264.7 macrophages and pma-induced mouse ear edema. Indian Journal of Pharmaceutical Sciences, 81(3), 533-538. https://doi.org/10.36468/pharmaceutical-sciences.540
  • Asgharpour, F., Moghadamnia, A. A., Motallebnejad, M., & Nouri, H. R. (2019). Propolis attenuates lipopolysaccharide-induced inflammatory responses through intracellular ROS and NO levels along with downregulation of IL-1β and IL-6 expressions in murine RAW 264.7 macrophages. Journal of food biochemistry, 43(8), e12926. https://doi.org/10.1111/jfbc.12926
  • Myint, T. M., Tezuka, Y., Midorikawa, K., & Matsushige, K. (2003). Inhibitors of nitric oxide (NO) production in murine macrophage-like J774. 1 cells from Brazilian propolis. Journal of traditional medicines, 20(1), 22-29.
There are 27 citations in total.

Details

Primary Language English
Subjects Food Engineering, Veterinary Surgery
Journal Section Research Articles
Authors

Serol Korkmaz 0000-0001-8970-6883

Ayşe Parmaksız 0000-0003-1242-7987

Ahmet Sait 0000-0001-7658-8793

Burcu İrem Omurtag Korkmaz 0000-0001-7918-6212

Project Number Yok
Publication Date April 30, 2022
Submission Date July 12, 2021
Acceptance Date April 12, 2022
Published in Issue Year 2022 Volume: 7 Issue: 1

Cite

APA Korkmaz, S., Parmaksız, A., Sait, A., Omurtag Korkmaz, B. İ. (2022). The analysis of the effects of propolis products as food supplements on the viability of baby hamster kidney (BHK-21) and murine macrophage (RAW 264.7) cells by spectrophotometric MTT assay. Journal of Advances in VetBio Science and Techniques, 7(1), 45-53. https://doi.org/10.31797/vetbio.970578

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