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Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells

Year 2023, Volume: 7 Issue: 2, 140 - 145, 15.11.2023
https://doi.org/10.30616/ajb.1343823

Abstract

The biological activities of lichens, known as organisms based on a symbiotic relationship, are attracting more and more attention in traditional medicine and modern drug research. Lichens can possess various pharmacological effects such as antimicrobial, antioxidant, antitumor, anti-inflammatory, and many others due to the bioactive compounds they contain. In the present study, Usnea longissima Ach. and its secondary metabolite, usnic acid on human gastric adenocarcinoma cells (AGS), human colorectal adenocarcinoma cells (Caco-2), and mouse fibroblasts (NIH/3T3) were investigated. In this context, methanol and water extracts from U. longissima were obtained by Soxhlet extractor. The characterization of usnic acid was carried out by fourier transform infrared spectroscopy (FTIR). The cytotoxic activities of the extracts and the metabolite on cells were determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) analysis. Considering the median inhibitory concentration (IC50) values, the application with the greater effect on AGS and NIH/3T3 cells was the methanol extract (373.17 µg/ml and 318.81 µg/ml, respectively). Considering the Caco-2 cells, it was determined that the water extract had the lowest IC50 value (230.05 µg/ml). The high cytotoxic activity of usnic acid on cancer cells (AGS; IC50: 395.03 µg/ml and Caco-2; IC50: 462.35 µg/ml) compared to normal cell (NIH/3T3; IC50: 472.41 µg/ml) was noted. As a result, it has been revealed that methanol and water extracts of U. longissima, especially usnic acid, are products that can be used within the scope of complementary therapy.

References

  • Adenubi OT, Famuyide IM, McGaw LJ, Eloff JN (2022). Lichens: an update on their ethnopharmacological uses and potential as sources of drug leads. Journal of Ethnopharmacology 298: 115657.
  • Bakar SZA, Latip J, Din L Bin, Samsuddin MW (2014). Metabolic fingerprinting of lichen usnea baileyi by fourier transform infrared spectroscopy. AIP Conference Proceedings 1614(February 2015): 452-455.
  • Bangalore PK, Pedapati RK, Pranathi AN, Batchu UR, Misra S, Estharala M, Sriram D, Kantevari S (2023). Aryl-n-hexanamide linked enaminones of usnic acid as promising antimicrobial agents. Molecular Diversity 27(2): 811-836.
  • Bézivin C, Tomasi S, Lohézic-Le Dévéhat F, Boustie J (2003). Cytotoxic activity of some lichen extracts on murine and human cancer cell lines. Phytomedicine 10(6-7): 499-503.
  • Bharti S, Nayaka S, Kumar R (2022). Evaluation of some traditional therapeutic properties of Usnea longissima (Ascomycota, lichenized fungi): antimicrobial, antiquorum and antioxidant. Journal of Microbiology, Biotechnology and Food Sciences 11(4): e3163.
  • Brisdelli F, Perilli M, Sellitri D, Piovano M, Garbarino JA, Nicoletti M, Bozzi A, Amicosante G, Celenza G (2013). Cytotoxic activity and antioxidant capacity of purified lichen metabolites: an in vitro study. Phytotherapy Research 27(3): 431-437.
  • Divya Reddy S, Siva B, Kumar K, Phani Babu VS, Sravanthi V, Boustie J, Lakshma Nayak V, Tiwari AK, Rao CHV, Sridhar B, Shashikala P, Suresh Babu K (2019). Comprehensive analysis of secondary metabolites in Usnea longissima (lichenized Ascomycetes, Parmeliaceae) using UPLC-ESI-QTOF-MS/MS and pro-apoptotic activity of barbatic acid. Molecules 24(12): 2270.
  • Emsen B, Aslan A (2018). Use of lichens as natural insecticide. Anatolian Journal of Botany 2(1): 22-27.
  • Emsen B, Aslan A, Turkez H, Taghizadehghalehjoughi A, Kaya A (2018). The anti‑cancer efficacies of diffractaic, lobaric, and usnic acid: in vitro inhibition of glioma. Journal of Cancer Research and Therapeutics 14(5): 941-951.
  • Esseen P-A, Ekström M (2023). Influence of canopy structure and light on the three-dimensional distribution of the iconic lichen Usnea longissima. Forest Ecology and Management 529: 120667.
  • Guo L, Shi Q, Fang J-L, Mei N, Ali AA, Lewis SM, Leakey JEA, Frankos VH (2008). Review of usnic acid and Usnea barbata toxicity. Journal of Environmental Science and Health, Part C 26(4): 317-338.
  • İnan B, Özçimen D (2021). Preparation and characterization of microalgal oil loaded alginate/poly (vinyl alcohol) electrosprayed nanoparticles. Food and Bioproducts Processing 129: 105-114.
  • Karabacak RB, Tay T, Kivanç M (2014). Preparation of novel antimicrobial polymer colloids based on (+)-usnic acid and poly (vinylbenzyl chloride). Reactive and Functional Polymers 83: 7-13.
  • Manojlović NT, Rančić AB, Décor R, Vasiljević P, Tomović J (2021). Determination of chemical composition and antimicrobial, antioxidant and cytotoxic activities of lichens Parmelia conspersa and Parmelia perlata. Journal of Food Measurement and Characterization 15(1): 686-696.
  • Maulidiyah M, Rachman F, Mulkiyan LOMZ, Natsir M, Nohong N, Darmawan A, Salim LOA, Nurdin M (2023). Antioxidant activity of usnic acid compound from methanol extract of lichen Usnea sp. Journal of Oleo Science 72(2): 179-188.
  • Miah R, Voon KWK, Huq AKMM, Rullah K, Tajuddin SN, Hamid HA, Aluwi MFFM (2023). Pharmacophore-based molecular docking of usnic acid derivatives to discover anti-viral drugs against influenza A virus. Journal of Research in Pharmacy 27(3): 1021-1038.
  • Mitrović T, Stamenković S, Cvetković V, Tošić S, Stanković M, Radojević I, Stefanović O, Comić L, Dačić D, Curčić M, Marković S (2011). Antioxidant, antimicrobial and antiproliferative activities of five lichen species. International Journal of Molecular Sciences 12(8): 5428-5448.
  • Nguyen TTH, Dinh MH, Chi HT, Wang SL, Nguyen QV, Tran TD, Nguyen AD (2019). Antioxidant and cytotoxic activity of lichens collected from Bidoup Nui Ba National Park, Vietnam. Research on Chemical Intermediates 45(1): 33-49.
  • Popovici V, Bucur L, Gîrd CE, Popescu A, Matei E, Cozaru GC, Schröder V, Ozon EA, Fița AC, Lupuliasa D, Botnarciuc M, Badea V (2022). Phenolic secondary metabolites and antiradical and antibacterial activities of different extracts of Usnea barbata (L.) Weber ex F.H.Wigg from Călimani Mountains, Romania. Pharmaceuticals 15(7): 829.
  • Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM (1992). The lichen flora of Great Britain and Ireland. In: The Lichen Flora of Great Britain and Ireland. London: Natural History Museum Publications in Association with the British Lichen Society.
  • Rajendran K, Karuppiah P, Ponnusamy P, Shaik MR, Khan M, Oh TH, Shaik B (2023a). Anti-inflammatory activity of mycobiont extract of Parmotrema austrosinense (Zahlbr.) Hale in a zebrafish model. Journal of Marine Science and Engineering 11(5): 1081.
  • Rajendran K, Ponmurugan P, Gnanamangai BM, Karuppiah P, Shaik MR, Khan M, Khan M, Shaik B (2023b). Bioefficacy of lecanoric acid produced by Parmotrema austrosinense (Zahlbr.) Hale against tea fungal pathogens. Horticulturae 9(6): 705.
  • Rajeswari N, Mesta AR, Kanivebagilu VS, Ramesh Babu HN (2019). Medicinal importance of usneoid lichens in Western Ghats, Southernindia. Plant Archives 19(2): 2540-2542.
  • Ranković B, Kosanić M (2019). Lichens as a potential source of bioactive secondary metabolites. In: Ranković B (ed) Lichen Secondary Metabolites. Springer, pp 1-29.
  • Roehm NW, Rodgers GH, Hatfield SM, Glasebrook AL (1991). An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. Journal of Immunological Methods 142(2): 257-265.
  • Somphong A, Poengsungnoen V, Buaruang K, Sripreechasak P, Khantasup K, Intaraudom C, Pittayakhajonwut P, Tanasupawat S, Phongsopitanun W (2023). The lichen-derived Streptomyces isolated from Pyxine cocoes produces the antibiotic with potent antimicrobial and antitumor activities. ScienceAsia 49(3): 328-336.
  • Stoica Oprea AE, Albuleț D, Bîrcă AC, Iordache F, Ficai A, Grumezescu AM, Vasile B Ș., Andronescu E, Marinescu F, Holban AM (2023). Electrospun nanofibrous mesh based on PVA, chitosan, and usnic acid for applications in wound healing. International Journal of Molecular Sciences 24(13): 11037.
  • Storaunet KO, Rolstad J, Rolstad E (2014). Effects of logging on the threatened epiphytic lichen Usnea longissima: an experimental approach. Silva Fennica 48(1): 949.
  • Studzińska-Sroka E, Tomczak H, Malińska N, Marta Wrońska, Kleszcz R, Galanty A, Cielecka-Piontek J, Latek D, Jarosław Paluszczak (2019). Cladonia uncialis as a valuable raw material of biosynthetic compounds against clinical strains of bacteria and fungi. Acta Biochimica Polonica 66(4): 1-7.
  • Şahin N, Emsen B, Aslan A, Sadi G (2021). Antioxidant potential of Pseudevernia furfuracea (L.) Zopf and its secondary metabolites on hepatocellular carcinoma cells: regulation of antioxidant enzymes. Anatolian Journal of Botany 5(2): 127-133.
  • Ureña-Vacas I, González-Burgos E, Divakar PK, Gómez-Serranillos MP (2022). Lichen depsidones with biological interest. Planta Medica 88(11): 855-880.
  • Wirth V (1995). Die Flechten Baden Württembergs. In: Die Flechten Baden Württembergs. Ulmer, Stuttgart, p 1006.
  • Yang Q, Song Z, Li X, Hou Y, Xu T, Wu S (2023). Lichen-derived actinomycetota: novel taxa and bioactive metabolites. International Journal of Molecular Sciences 24(8): 7341.

Usnea longissima Ach. ekstraktları ve onun sekonder metaboliti usnik asidin farklı hücreler üzerindeki sitotoksisitesi

Year 2023, Volume: 7 Issue: 2, 140 - 145, 15.11.2023
https://doi.org/10.30616/ajb.1343823

Abstract

Simbiyotik bir ilişkiye dayanan organizmalar olarak bilinen likenlerin biyolojik aktiviteleri, geleneksel tıp ve modern ilaç araştırmalarında giderek daha fazla ilgi çekmektedir. Likenler, içerdikleri biyoaktif bileşikler sayesinde antimikrobiyal, antioksidan, antitümör, antiinflamatuar ve diğer pek çok farmakolojik etkiye sahip olabilir. Mevcut çalışmada, Usnea longissima Ach. ve onun sekonder metaboliti usnik asidin insan gastrik adenokarsinom hücreleri (AGS), insan kolorektal adenokarsinom hücreleri (Caco-2) ve fare fibroblastları (NIH/3T3) üzerindeki sitotoksik etkileri incelenmiştir. Bu kapsamda, U. longissima’dan metanol ve su ekstraktları Soxhlet ekstraktörü ile elde edilmiştir. Usnik asidin karakterizasyonu fourier dönüşümlü kizilötesi spektroskopisi (FTIR) ile gerçekleştirilmiştir. Ekstraktlar ve metabolitin hücreler üzerindeki sitotoksik aktiviteleri 2,3-bis-(2-metoksi-4-nitro-5-sülfofenil)-2H-tetrazolyum-5-karboksanilid (XTT) analizi ile tespit edilmiştir. Medyan inhibitör konsantrasyonu (IC50) değerleri dikkate alındığında, AGS ve NIH/3T3 hücreleri üzerinde daha fazla etkili olan uygulama metanol ekstraktı olmuştur (sırasıyla, 373.17 µg/ml ve 318.81 µg/ml). Caco-2 hücreleri dikkate alındığında, su ekstraktının en düşük IC50 değerine (230.05 µg/ml) sahip olduğu belirlenmiştir. Usnik asidin normal hücreye (NIH/3T3; IC50: 472.41 µg/ml) kıyasla kanserli hücreler (AGS; IC50: 395.03 µg/ml ve Caco-2; IC50: 462.35 µg/ml) üzerindeki yüksek sitotoksik etkinliği dikkat çekmiştir. Sonuç olarak, başta usnik asit olmak üzere U. longissima'nın metanol ve su ekstraktlarının tamamlayıcı tedavi kapsamında kullanılabilecek ürünler olduğu ortaya çıkmıştır.

References

  • Adenubi OT, Famuyide IM, McGaw LJ, Eloff JN (2022). Lichens: an update on their ethnopharmacological uses and potential as sources of drug leads. Journal of Ethnopharmacology 298: 115657.
  • Bakar SZA, Latip J, Din L Bin, Samsuddin MW (2014). Metabolic fingerprinting of lichen usnea baileyi by fourier transform infrared spectroscopy. AIP Conference Proceedings 1614(February 2015): 452-455.
  • Bangalore PK, Pedapati RK, Pranathi AN, Batchu UR, Misra S, Estharala M, Sriram D, Kantevari S (2023). Aryl-n-hexanamide linked enaminones of usnic acid as promising antimicrobial agents. Molecular Diversity 27(2): 811-836.
  • Bézivin C, Tomasi S, Lohézic-Le Dévéhat F, Boustie J (2003). Cytotoxic activity of some lichen extracts on murine and human cancer cell lines. Phytomedicine 10(6-7): 499-503.
  • Bharti S, Nayaka S, Kumar R (2022). Evaluation of some traditional therapeutic properties of Usnea longissima (Ascomycota, lichenized fungi): antimicrobial, antiquorum and antioxidant. Journal of Microbiology, Biotechnology and Food Sciences 11(4): e3163.
  • Brisdelli F, Perilli M, Sellitri D, Piovano M, Garbarino JA, Nicoletti M, Bozzi A, Amicosante G, Celenza G (2013). Cytotoxic activity and antioxidant capacity of purified lichen metabolites: an in vitro study. Phytotherapy Research 27(3): 431-437.
  • Divya Reddy S, Siva B, Kumar K, Phani Babu VS, Sravanthi V, Boustie J, Lakshma Nayak V, Tiwari AK, Rao CHV, Sridhar B, Shashikala P, Suresh Babu K (2019). Comprehensive analysis of secondary metabolites in Usnea longissima (lichenized Ascomycetes, Parmeliaceae) using UPLC-ESI-QTOF-MS/MS and pro-apoptotic activity of barbatic acid. Molecules 24(12): 2270.
  • Emsen B, Aslan A (2018). Use of lichens as natural insecticide. Anatolian Journal of Botany 2(1): 22-27.
  • Emsen B, Aslan A, Turkez H, Taghizadehghalehjoughi A, Kaya A (2018). The anti‑cancer efficacies of diffractaic, lobaric, and usnic acid: in vitro inhibition of glioma. Journal of Cancer Research and Therapeutics 14(5): 941-951.
  • Esseen P-A, Ekström M (2023). Influence of canopy structure and light on the three-dimensional distribution of the iconic lichen Usnea longissima. Forest Ecology and Management 529: 120667.
  • Guo L, Shi Q, Fang J-L, Mei N, Ali AA, Lewis SM, Leakey JEA, Frankos VH (2008). Review of usnic acid and Usnea barbata toxicity. Journal of Environmental Science and Health, Part C 26(4): 317-338.
  • İnan B, Özçimen D (2021). Preparation and characterization of microalgal oil loaded alginate/poly (vinyl alcohol) electrosprayed nanoparticles. Food and Bioproducts Processing 129: 105-114.
  • Karabacak RB, Tay T, Kivanç M (2014). Preparation of novel antimicrobial polymer colloids based on (+)-usnic acid and poly (vinylbenzyl chloride). Reactive and Functional Polymers 83: 7-13.
  • Manojlović NT, Rančić AB, Décor R, Vasiljević P, Tomović J (2021). Determination of chemical composition and antimicrobial, antioxidant and cytotoxic activities of lichens Parmelia conspersa and Parmelia perlata. Journal of Food Measurement and Characterization 15(1): 686-696.
  • Maulidiyah M, Rachman F, Mulkiyan LOMZ, Natsir M, Nohong N, Darmawan A, Salim LOA, Nurdin M (2023). Antioxidant activity of usnic acid compound from methanol extract of lichen Usnea sp. Journal of Oleo Science 72(2): 179-188.
  • Miah R, Voon KWK, Huq AKMM, Rullah K, Tajuddin SN, Hamid HA, Aluwi MFFM (2023). Pharmacophore-based molecular docking of usnic acid derivatives to discover anti-viral drugs against influenza A virus. Journal of Research in Pharmacy 27(3): 1021-1038.
  • Mitrović T, Stamenković S, Cvetković V, Tošić S, Stanković M, Radojević I, Stefanović O, Comić L, Dačić D, Curčić M, Marković S (2011). Antioxidant, antimicrobial and antiproliferative activities of five lichen species. International Journal of Molecular Sciences 12(8): 5428-5448.
  • Nguyen TTH, Dinh MH, Chi HT, Wang SL, Nguyen QV, Tran TD, Nguyen AD (2019). Antioxidant and cytotoxic activity of lichens collected from Bidoup Nui Ba National Park, Vietnam. Research on Chemical Intermediates 45(1): 33-49.
  • Popovici V, Bucur L, Gîrd CE, Popescu A, Matei E, Cozaru GC, Schröder V, Ozon EA, Fița AC, Lupuliasa D, Botnarciuc M, Badea V (2022). Phenolic secondary metabolites and antiradical and antibacterial activities of different extracts of Usnea barbata (L.) Weber ex F.H.Wigg from Călimani Mountains, Romania. Pharmaceuticals 15(7): 829.
  • Purvis OW, Coppins BJ, Hawksworth DL, James PW, Moore DM (1992). The lichen flora of Great Britain and Ireland. In: The Lichen Flora of Great Britain and Ireland. London: Natural History Museum Publications in Association with the British Lichen Society.
  • Rajendran K, Karuppiah P, Ponnusamy P, Shaik MR, Khan M, Oh TH, Shaik B (2023a). Anti-inflammatory activity of mycobiont extract of Parmotrema austrosinense (Zahlbr.) Hale in a zebrafish model. Journal of Marine Science and Engineering 11(5): 1081.
  • Rajendran K, Ponmurugan P, Gnanamangai BM, Karuppiah P, Shaik MR, Khan M, Khan M, Shaik B (2023b). Bioefficacy of lecanoric acid produced by Parmotrema austrosinense (Zahlbr.) Hale against tea fungal pathogens. Horticulturae 9(6): 705.
  • Rajeswari N, Mesta AR, Kanivebagilu VS, Ramesh Babu HN (2019). Medicinal importance of usneoid lichens in Western Ghats, Southernindia. Plant Archives 19(2): 2540-2542.
  • Ranković B, Kosanić M (2019). Lichens as a potential source of bioactive secondary metabolites. In: Ranković B (ed) Lichen Secondary Metabolites. Springer, pp 1-29.
  • Roehm NW, Rodgers GH, Hatfield SM, Glasebrook AL (1991). An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. Journal of Immunological Methods 142(2): 257-265.
  • Somphong A, Poengsungnoen V, Buaruang K, Sripreechasak P, Khantasup K, Intaraudom C, Pittayakhajonwut P, Tanasupawat S, Phongsopitanun W (2023). The lichen-derived Streptomyces isolated from Pyxine cocoes produces the antibiotic with potent antimicrobial and antitumor activities. ScienceAsia 49(3): 328-336.
  • Stoica Oprea AE, Albuleț D, Bîrcă AC, Iordache F, Ficai A, Grumezescu AM, Vasile B Ș., Andronescu E, Marinescu F, Holban AM (2023). Electrospun nanofibrous mesh based on PVA, chitosan, and usnic acid for applications in wound healing. International Journal of Molecular Sciences 24(13): 11037.
  • Storaunet KO, Rolstad J, Rolstad E (2014). Effects of logging on the threatened epiphytic lichen Usnea longissima: an experimental approach. Silva Fennica 48(1): 949.
  • Studzińska-Sroka E, Tomczak H, Malińska N, Marta Wrońska, Kleszcz R, Galanty A, Cielecka-Piontek J, Latek D, Jarosław Paluszczak (2019). Cladonia uncialis as a valuable raw material of biosynthetic compounds against clinical strains of bacteria and fungi. Acta Biochimica Polonica 66(4): 1-7.
  • Şahin N, Emsen B, Aslan A, Sadi G (2021). Antioxidant potential of Pseudevernia furfuracea (L.) Zopf and its secondary metabolites on hepatocellular carcinoma cells: regulation of antioxidant enzymes. Anatolian Journal of Botany 5(2): 127-133.
  • Ureña-Vacas I, González-Burgos E, Divakar PK, Gómez-Serranillos MP (2022). Lichen depsidones with biological interest. Planta Medica 88(11): 855-880.
  • Wirth V (1995). Die Flechten Baden Württembergs. In: Die Flechten Baden Württembergs. Ulmer, Stuttgart, p 1006.
  • Yang Q, Song Z, Li X, Hou Y, Xu T, Wu S (2023). Lichen-derived actinomycetota: novel taxa and bioactive metabolites. International Journal of Molecular Sciences 24(8): 7341.
There are 33 citations in total.

Details

Primary Language English
Subjects Biochemistry and Cell Biology (Other)
Journal Section Articles
Authors

Tubanur Aslan Engin 0000-0003-2885-8829

Buğrahan Emsen 0000-0002-9636-2596

Rabia Yılmaz 0000-0002-1896-2932

Rabia Cakir Koc 0000-0002-8545-9878

Benan İnan 0000-0002-2315-3099

Didem Ozcimen 0000-0003-2483-7617

Early Pub Date September 14, 2023
Publication Date November 15, 2023
Acceptance Date September 3, 2023
Published in Issue Year 2023 Volume: 7 Issue: 2

Cite

APA Aslan Engin, T., Emsen, B., Yılmaz, R., Cakir Koc, R., et al. (2023). Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Anatolian Journal of Botany, 7(2), 140-145. https://doi.org/10.30616/ajb.1343823
AMA Aslan Engin T, Emsen B, Yılmaz R, Cakir Koc R, İnan B, Ozcimen D. Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Ant J Bot. November 2023;7(2):140-145. doi:10.30616/ajb.1343823
Chicago Aslan Engin, Tubanur, Buğrahan Emsen, Rabia Yılmaz, Rabia Cakir Koc, Benan İnan, and Didem Ozcimen. “Cytotoxicity of Usnea Longissima Ach. Extracts and Its Secondary Metabolite, Usnic Acid on Different Cells”. Anatolian Journal of Botany 7, no. 2 (November 2023): 140-45. https://doi.org/10.30616/ajb.1343823.
EndNote Aslan Engin T, Emsen B, Yılmaz R, Cakir Koc R, İnan B, Ozcimen D (November 1, 2023) Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Anatolian Journal of Botany 7 2 140–145.
IEEE T. Aslan Engin, B. Emsen, R. Yılmaz, R. Cakir Koc, B. İnan, and D. Ozcimen, “Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells”, Ant J Bot, vol. 7, no. 2, pp. 140–145, 2023, doi: 10.30616/ajb.1343823.
ISNAD Aslan Engin, Tubanur et al. “Cytotoxicity of Usnea Longissima Ach. Extracts and Its Secondary Metabolite, Usnic Acid on Different Cells”. Anatolian Journal of Botany 7/2 (November 2023), 140-145. https://doi.org/10.30616/ajb.1343823.
JAMA Aslan Engin T, Emsen B, Yılmaz R, Cakir Koc R, İnan B, Ozcimen D. Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Ant J Bot. 2023;7:140–145.
MLA Aslan Engin, Tubanur et al. “Cytotoxicity of Usnea Longissima Ach. Extracts and Its Secondary Metabolite, Usnic Acid on Different Cells”. Anatolian Journal of Botany, vol. 7, no. 2, 2023, pp. 140-5, doi:10.30616/ajb.1343823.
Vancouver Aslan Engin T, Emsen B, Yılmaz R, Cakir Koc R, İnan B, Ozcimen D. Cytotoxicity of Usnea longissima Ach. extracts and its secondary metabolite, usnic acid on different cells. Ant J Bot. 2023;7(2):140-5.

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