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Farklı Doz ve Boyuttaki BSA Nanopartiküllerinin Ankara Tekesi Spermasının Kriyoprezervasyonu Üzerindeki Etkileri: Motilite, Membranın Fonksiyonel Bütünlüğü ve Canlılık

Year 2025, Volume: 18 Issue: 3, 395 - 402
https://doi.org/10.46309/biodicon.2025.1630745

Abstract

Amaç: BSA, 585 amino asit içeren ve yüksek oranda çözünebilir bir madde olup, uzun zamandır farklı türlerde spermanın dondurulmasında başarıyla kullanılmaktadır. Ancak, BSA'nın nanoformları (BSA NPs), yüksek biyoyararlanım kapasiteleri ve artan yüzey alanları sayesinde son yıllarda farklı hücrelerde ilaç taşıma uygulamalarında kullanılmaktadır. Bu çalışmada, BSA NPs'ın Ankara tekesi spermasının dondurulabilirliği üzerindeki etkileri incelenmiştir.
Metod: Beş farklı Ankara tekesi sperması, 6 tekrar halinde toplanmış ve kontrol, BSA 20-5, BSA 20-10, BSA 40-5 ve BSA 40-10 olacak şekilde 5 gruba ayrılmıştır. Sıvı azot buharında dondurulan spermalar, 37 ºC'de çözdürülmüş ve motilite, membranın fonksiyonel bütünlüğü (HOST) ve canlı/ölü oranları değerlendirilmiştir.
Bulgular: Elde edilen veriler, motilite ve canlı/ölü oranı açısından gruplar arasında istatistiksel olarak anlamlı bir fark olmadığını göstermektedir. Ancak, HOST testi sonucunda, kontrol grubu ve BSA 20-5 grubu diğer gruplara oranla daha yüksek membran dayanım kabiliyetine sahip bulunurken, BSA 20-10, BSA 40-5 ve BSA 40-10 gruplarındaki spermaların membran dayanım kabiliyeti düşmüştür. Bu durumun, BSA NPs'ın membrandaki kolesterolü delokalize etmesinden kaynaklandığı düşünülmektedir.
Sonuç: Sonuç olarak, düşük doz ve boyuttaki BSA NPs'ın spermaya zarar vermediği ve ilaç taşıma sistemlerinde kullanım potansiyeli taşıdığı sonucuna varılmıştır.

Ethical Statement

Acknowledgement: We would like to thank Erciyes University Scientific Research Projects Coordination Unit for supporting the project with the number TSA-2022-12015. We would also like to thank the Directorate of Selcuk University Veterinary Faculty, Humeyra Özgen Research and Application Farm, where the Angora bucks used in the project were maintained and fed. Conflicts of interest: The authors of this study declare that they have no conflicts of interest with any institutions or organizations. Funding: This project was funded by the Erciyes University Scientific Research Projects Coordination Unit under project number TSA-2022-12015. Ethical statement: The ethical approval for this study was granted by the Ethics Committee of Selçuk University, Faculty of Veterinary Medicine, Animal Research and Production Center, with decision number 2022/40 dated 06.05.2022. Author contributions: Ali Erdem ÖZTÜRK, Yunus Emre ATAY and Mustafa BODU designed the research study. Ali Erdem ÖZTÜRK, Mustafa BODU and Zeliha KILINÇ performed the research. Serpil SARIÖZKAN collected and analyzed the data. İsmail ÖÇSOY has been involved in drafting the manuscript. All authors give final approval of the version to be published. This study was presented as an oral presentation at the conference titled ‘Elegans International Scientific Conference Series, First International Conference on Life Science’ and its abstract was published in the conference book.

Supporting Institution

Erciyes University Scientific Research Projects Coordination Unit

Project Number

TSA-2022-12015

Thanks

We would like to thank Erciyes University Scientific Research Projects Coordination Unit for supporting the project with the number TSA-2022-12015. We would also like to thank the Directorate of Selcuk University Veterinary Faculty Humeyra Özgen Research and Application Farm where the ankara goats used in the project were maintained and fed.

References

  • [1] Roche, M., Rondeau, P., Singh, N. R., Tarnus, E., & Bourdon, E. (2008). The antioxidant properties of serum albumin. FEBS letters, 582(13), 1783-1787. https://doi.org/10.1016/j.febslet.2008.04.057
  • [2] Peters, T. (1996). All about albumin. San Diego, CA: Academic Press.
  • [3] Bourdon, E., & Blache, D. (2001). The importance of proteins in defense against oxidation. Antioxidants and Redox Signaling, 3(2), 293-311. https://doi.org/10.1089/152308601300185241
  • [4] Halliwell, B. (1988). Albumin—an important extracellular antioxidant?. Biochemical pharmacology, 37(4), 569-571. https://doi.org/10.1016/0006-2952(88)90126-8
  • [5] Quinlan, G. J., Martin, G. S., & Evans, T. W. (2005). Albumin: biochemical properties and therapeutic potential. Hepatology, 41(6), 1211-1219. Doi: 10.1002/hep.20720
  • [6] Fukuzawa, K., Saitoh, Y., Akai, K., Kogure, K., Ueno, S., Tokumura, A., ... & Shibata, A. (2005). Antioxidant effect of bovine serum albumin on membrane lipid peroxidation induced by iron chelate and superoxide. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1668(1), 145-155. https://doi.org/10.1016/j.bbamem.2004.12.006
  • [7] Prien, S., & Iacovides, S. (2016). Cryoprotectants & cryopreservation of equine semen: A review of industry cryoprotectants and the effects of cryopreservation on equine semen membranes. J. Dairy Vet. Anim. Res, 3(1), 1-8. https://doi.org/10.1016/j.anireprosci.2008.03.016
  • [8] Vidament, M., Vincent, P., Martin, F. X., Magistrini, M., & Blesbois, E. (2009). Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Animal Reproduction Science, 112(1-2), 22-35. https://doi.org/10.1016/j.anireprosci.2008.03.016
  • [9] Montoya Páez, J. D., Úsuga Suarez, A., & Restrepo Betancur, G. (2023). Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma. Reproduction in Domestic Animals, 58(4), 486-495. https://doi.org/10.1111/rda.14309
  • [10] Galisteo-González, F., & Molina-Bolívar, J. A. (2014). Systematic study on the preparation of BSA nanoparticles. Colloids and Surfaces B: Biointerfaces, 123, 286-292. https://doi.org/10.1016/j.colsurfb.2014.09.028
  • [11] Pedrozo, R. C., Oliveira, L. F., Klein, T., Ferreira, L. M. B., & Beck, R. C. R. (2020). Bovine serum albumin-based nanoparticles containing the flavonoid rutin produced by nano spray drying. Brazilian Journal of Pharmaceutical Sciences, 56, e17692. https://doi.org/10.1590/s2175-97902019000317692
  • [12] Evans, G. (1987). Salamon's artificial insemination of sheep and goats. London, UK: Butterworths.
  • [13] Takahashi, K., Uchida, A., & Kitao, M. (1990). Hypoosmotic swelling test of sperm. Archives of Andrology, 25(3), 225-242. https://doi.org/10.3109/01485019008987627 [14] Swanson, E. W., & Bearden, H. J. (1951). An eosin-nigrosin stain for differentiating live and dead bovine spermatozoa. Journal of Animal Science, 10(4), 981-987. https://doi.org/10.2527/jas1951.104981x
  • [15] Yu, Z., Xu, H., Kawai, T., Kamei, N., & Fukuda, M. (2014). Bovine serum albumin nanoparticles as controlled release carrier for local drug delivery to the inner ear. Nanoscale Research Letters, 9, 224. https://doi.org/10.1186/1556-276X-9-224
  • [16] Naijian, H. R., Kohram, H., Shahneh, A. Z., Shakeri, M., & Zhandi, M. (2013). Effects of different concentrations of BHT on microscopic and oxidative parameters of Mahabadi goat semen following the freeze–thaw process. Cryobiology, 66(2), 151-155. https://doi.org/10.1016/j.cryobiol.2012.12.003
  • [17] Rayan, M., Anwer, A., El-Badry, D., & Zain, A. E. D. (2023). The Effects of Bovine Serum Albumin (BSA) Supplementation on Post-thaw Quality of Cryopreserved Bull Semen. مجلة سوهاج لشباب الباحثين, 3(3), 43-54.‎ https://doi.org/10.21608/sjyr.2023.302847
  • [18] Yamashiro, H., et al., Enhanced freezability of goat spermatozoa collected into tubes containing extender supplemented with bovine serum albumin (BSA). Journal of Reproduction and Development, 2006. 52(3): p. 407-414. https://doi.org/10.1262/jrd.17105
  • [19] Uysal, O., T. Korkmaz, and H. Tosun, Effect of bovine serum albumin on freezing of canine semen. 2005.
  • [20] Uysal, O., & Bucak, M. N. (2007). Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Veterinaria Brno, 76(3), 383-390. https://doi.org/10.2754/avb200776030383
  • [21] Sarıözkan, S., et al., The effect of bovine serum albumin and fetal calf serum on sperm quality, DNA fragmentation and lipid peroxidation of the liquid stored rabbit semen. Cryobiology, 2013. 67(1): p. 1-6. https://doi.org/10.1016/j.cryobiol.2013.04.002
  • [22] El-Kon, I., Testing usability of bovine serum albumin (BSA) for preservation of Egyptian buffalo semen. American-Eurasian Journal of Agricultural & Environmental Sciences, 2011. 11(4): p. 495-502.
  • [23] Krause, M. R., & Regen, S. L. (2014). The structural role of cholesterol in cell membranes: from condensed bilayers to lipid rafts. Accounts of chemical research, 47(12), 3512-3521. https://doi.org/10.1021/ar500260t
  • [24] Yeagle, P. L. (1985). Cholesterol and the cell membrane. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes, 822(3-4), 267-287. https://doi.org/10.1016/0304-4157(85)90011-5
  • [25] Sankaranarayanan, S., de la Llera-Moya, M., Drazul-Schrader, D., Phillips, M. C., Kellner-Weibel, G., & Rothblat, G. H. (2013). Serum albumin acts as a shuttle to enhance cholesterol efflux from cells. Journal of lipid research, 54(3), 671-676. https://doi.org/10.1194/jlr.M031336

Effects of different doses and sizes of BSA nanoparticles on cryopreservation of angora buck semen: motility, functional membrane ıntegrity and, viability

Year 2025, Volume: 18 Issue: 3, 395 - 402
https://doi.org/10.46309/biodicon.2025.1630745

Abstract

Purpose: BSA is a highly soluble substance consisting of 585 amino acids, which has been successfully used for the cryopreservation of sperm from various species for a long time. However, the nanoforms of BSA (BSA NPs) have been utilized in drug delivery applications in different cell types in recent years due to their high bioavailability and increased surface areas. In this study, we investigate the effects of BSA NPs on the cryopreservability of Angora buck sperm.
Method: Sperm samples from five different Ankara rams were collected in six replicates and divided into five groups: control, BSA 20-5, BSA 20-10, BSA 40-5, and BSA 40-10. The sperm, frozen in liquid nitrogen vapor, were thawed at 37 ºC, and motility, functional membrane integrity (HOST), and live/dead ratios were assessed.
Findings: The data obtained showed no statistically significant difference among the groups in terms of motility and live/dead ratios. However, in the HOST test, the control group and the BSA 20-5 group had higher resistance capabilities compared to the other groups, while BSA 20-10, BSA 40-5, and BSA 40-10 groups reduced membrane resistance capability. This situation is considered to result from the removal of cholesterol from the membrane by BSA nanoparticles.
Conclusion: Consequently, the study concludes that BSA NPs used at low doses and sizes do not harm sperm and hold promise for use in drug delivery systems.

Ethical Statement

Acknowledgement: We would like to thank Erciyes University Scientific Research Projects Coordination Unit for supporting the project with the number TSA-2022-12015. We would also like to thank the Directorate of Selcuk University Veterinary Faculty, Humeyra Özgen Research and Application Farm, where the Angora bucks used in the project were maintained and fed. Conflicts of interest: The authors of this study declare that they have no conflicts of interest with any institutions or organizations. Funding: This project was funded by the Erciyes University Scientific Research Projects Coordination Unit under project number TSA-2022-12015. Ethical statement: The ethical approval for this study was granted by the Ethics Committee of Selçuk University, Faculty of Veterinary Medicine, Animal Research and Production Center, with decision number 2022/40 dated 06.05.2022. Author contributions: Ali Erdem ÖZTÜRK, Yunus Emre ATAY and Mustafa BODU designed the research study. Ali Erdem ÖZTÜRK, Mustafa BODU and Zeliha KILINÇ performed the research. Serpil SARIÖZKAN collected and analyzed the data. İsmail ÖÇSOY has been involved in drafting the manuscript. All authors give final approval of the version to be published. This study was presented as an oral presentation at the conference titled ‘Elegans International Scientific Conference Series, First International Conference on Life Science’ and its abstract was published in the conference book.

Project Number

TSA-2022-12015

References

  • [1] Roche, M., Rondeau, P., Singh, N. R., Tarnus, E., & Bourdon, E. (2008). The antioxidant properties of serum albumin. FEBS letters, 582(13), 1783-1787. https://doi.org/10.1016/j.febslet.2008.04.057
  • [2] Peters, T. (1996). All about albumin. San Diego, CA: Academic Press.
  • [3] Bourdon, E., & Blache, D. (2001). The importance of proteins in defense against oxidation. Antioxidants and Redox Signaling, 3(2), 293-311. https://doi.org/10.1089/152308601300185241
  • [4] Halliwell, B. (1988). Albumin—an important extracellular antioxidant?. Biochemical pharmacology, 37(4), 569-571. https://doi.org/10.1016/0006-2952(88)90126-8
  • [5] Quinlan, G. J., Martin, G. S., & Evans, T. W. (2005). Albumin: biochemical properties and therapeutic potential. Hepatology, 41(6), 1211-1219. Doi: 10.1002/hep.20720
  • [6] Fukuzawa, K., Saitoh, Y., Akai, K., Kogure, K., Ueno, S., Tokumura, A., ... & Shibata, A. (2005). Antioxidant effect of bovine serum albumin on membrane lipid peroxidation induced by iron chelate and superoxide. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1668(1), 145-155. https://doi.org/10.1016/j.bbamem.2004.12.006
  • [7] Prien, S., & Iacovides, S. (2016). Cryoprotectants & cryopreservation of equine semen: A review of industry cryoprotectants and the effects of cryopreservation on equine semen membranes. J. Dairy Vet. Anim. Res, 3(1), 1-8. https://doi.org/10.1016/j.anireprosci.2008.03.016
  • [8] Vidament, M., Vincent, P., Martin, F. X., Magistrini, M., & Blesbois, E. (2009). Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Animal Reproduction Science, 112(1-2), 22-35. https://doi.org/10.1016/j.anireprosci.2008.03.016
  • [9] Montoya Páez, J. D., Úsuga Suarez, A., & Restrepo Betancur, G. (2023). Donkey semen cryopreservation: Alternatives with permeable, non‐permeable cryoprotectants and seminal plasma. Reproduction in Domestic Animals, 58(4), 486-495. https://doi.org/10.1111/rda.14309
  • [10] Galisteo-González, F., & Molina-Bolívar, J. A. (2014). Systematic study on the preparation of BSA nanoparticles. Colloids and Surfaces B: Biointerfaces, 123, 286-292. https://doi.org/10.1016/j.colsurfb.2014.09.028
  • [11] Pedrozo, R. C., Oliveira, L. F., Klein, T., Ferreira, L. M. B., & Beck, R. C. R. (2020). Bovine serum albumin-based nanoparticles containing the flavonoid rutin produced by nano spray drying. Brazilian Journal of Pharmaceutical Sciences, 56, e17692. https://doi.org/10.1590/s2175-97902019000317692
  • [12] Evans, G. (1987). Salamon's artificial insemination of sheep and goats. London, UK: Butterworths.
  • [13] Takahashi, K., Uchida, A., & Kitao, M. (1990). Hypoosmotic swelling test of sperm. Archives of Andrology, 25(3), 225-242. https://doi.org/10.3109/01485019008987627 [14] Swanson, E. W., & Bearden, H. J. (1951). An eosin-nigrosin stain for differentiating live and dead bovine spermatozoa. Journal of Animal Science, 10(4), 981-987. https://doi.org/10.2527/jas1951.104981x
  • [15] Yu, Z., Xu, H., Kawai, T., Kamei, N., & Fukuda, M. (2014). Bovine serum albumin nanoparticles as controlled release carrier for local drug delivery to the inner ear. Nanoscale Research Letters, 9, 224. https://doi.org/10.1186/1556-276X-9-224
  • [16] Naijian, H. R., Kohram, H., Shahneh, A. Z., Shakeri, M., & Zhandi, M. (2013). Effects of different concentrations of BHT on microscopic and oxidative parameters of Mahabadi goat semen following the freeze–thaw process. Cryobiology, 66(2), 151-155. https://doi.org/10.1016/j.cryobiol.2012.12.003
  • [17] Rayan, M., Anwer, A., El-Badry, D., & Zain, A. E. D. (2023). The Effects of Bovine Serum Albumin (BSA) Supplementation on Post-thaw Quality of Cryopreserved Bull Semen. مجلة سوهاج لشباب الباحثين, 3(3), 43-54.‎ https://doi.org/10.21608/sjyr.2023.302847
  • [18] Yamashiro, H., et al., Enhanced freezability of goat spermatozoa collected into tubes containing extender supplemented with bovine serum albumin (BSA). Journal of Reproduction and Development, 2006. 52(3): p. 407-414. https://doi.org/10.1262/jrd.17105
  • [19] Uysal, O., T. Korkmaz, and H. Tosun, Effect of bovine serum albumin on freezing of canine semen. 2005.
  • [20] Uysal, O., & Bucak, M. N. (2007). Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Veterinaria Brno, 76(3), 383-390. https://doi.org/10.2754/avb200776030383
  • [21] Sarıözkan, S., et al., The effect of bovine serum albumin and fetal calf serum on sperm quality, DNA fragmentation and lipid peroxidation of the liquid stored rabbit semen. Cryobiology, 2013. 67(1): p. 1-6. https://doi.org/10.1016/j.cryobiol.2013.04.002
  • [22] El-Kon, I., Testing usability of bovine serum albumin (BSA) for preservation of Egyptian buffalo semen. American-Eurasian Journal of Agricultural & Environmental Sciences, 2011. 11(4): p. 495-502.
  • [23] Krause, M. R., & Regen, S. L. (2014). The structural role of cholesterol in cell membranes: from condensed bilayers to lipid rafts. Accounts of chemical research, 47(12), 3512-3521. https://doi.org/10.1021/ar500260t
  • [24] Yeagle, P. L. (1985). Cholesterol and the cell membrane. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes, 822(3-4), 267-287. https://doi.org/10.1016/0304-4157(85)90011-5
  • [25] Sankaranarayanan, S., de la Llera-Moya, M., Drazul-Schrader, D., Phillips, M. C., Kellner-Weibel, G., & Rothblat, G. H. (2013). Serum albumin acts as a shuttle to enhance cholesterol efflux from cells. Journal of lipid research, 54(3), 671-676. https://doi.org/10.1194/jlr.M031336
There are 24 citations in total.

Details

Primary Language English
Subjects Ecology (Other)
Journal Section Research Articles
Authors

Ali Erdem Öztürk 0000-0002-3053-4615

Mustafa Bodu 0000-0002-8432-7347

Serpil Sarıözkan 0000-0001-5224-2341

Yunus Emre Atay 0000-0002-8616-7068

Zeliha Kilinç 0009-0008-0788-8379

İsmail Öçsoy 0000-0002-5991-3934

Project Number TSA-2022-12015
Early Pub Date September 25, 2025
Publication Date October 6, 2025
Submission Date January 31, 2025
Acceptance Date March 23, 2025
Published in Issue Year 2025 Volume: 18 Issue: 3

Cite

APA Öztürk, A. E., Bodu, M., Sarıözkan, S., … Atay, Y. E. (2025). Effects of different doses and sizes of BSA nanoparticles on cryopreservation of angora buck semen: motility, functional membrane ıntegrity and, viability. Biological Diversity and Conservation, 18(3), 395-402. https://doi.org/10.46309/biodicon.2025.1630745

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