Assessment of lecithin nanoliposomes addition to fresh low-density lipoprotein-based diluent for cold storage of ram semen

Document Type : Full paper (Original article)

Authors

1 Graduated from Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

3 Department of Animal Sciences, Faculty of Agriculture, Urmia University, Urmia, Iran

10.22099/ijvr.2025.51361.7626

Abstract

Background: Maintaining the quality of collected semen is a crucial goal in assisted reproductive technologies. Aims: The present study aimed to evaluate the effect of lecithin nanoliposomes (LNLs) addition to purified low-density lipoproteins (LDL)-based extender on cold preservation of ram semen for up to 72 h. Methods: Ejaculates were collected from fertile rams, assessed, diluted with tris-citric acid-fructose-purified LDL extender, pooled and used within the experiment. Experimental groups supplemented with 0 (control), 1, 2, 4, and 8 mM, LNLs and stored at 4°C for 72 h. Kinematics, viability, DNA fragmentation index (DFI), membrane integrity (MI), malondialdehyde (MDA) and total nitrate nitrite (TNN) were assessed at 0, 24, 48 and 72 h. Results: Total motility, forward progressive motility, curvilinear velocity, average path velocity and straightness variables were improved by 1-8 mM LNLs addition (P<0.05). Greater viability (at 48 and 72 h) and MI (at 24, 48 and 72 h), and lower DFI (at 24, 48 and 72 h) were detected in LNLs treated groups compared with the control group (P<0.05). LNLs addition at 2-8 mM resulted in lower amounts of MDA and TNN at 48 and 72 h (P<0.05). Conclusion: LNLs were recommended as a protectant in ram semen tris-LDL-based diluent because they reduce peroxidative/nitrosative damage, fragmented DNA indices, and improve kinematics during cold preservation.

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References

Alvarez, JG and Storey, BT (1995). Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa. Mol. Reprod. Dev., 42: 334-346.
Alvarez-Rodríguez, M; Alvarez, M; Anel-Lópe, L; Martínez-Rodríguez, C; Martínez-Pastor, F; Borragan, S; Anel, L and De Paz, P (2013). The antioxidant effects of soybean lecithin-or low-density lipoprotein-based extenders for the cryopreservation of brown-bear (ursus arctos) spermatozoa. Reprod. Fertil. Dev., 25: 1185-1193.
Ashrafi, I; Kohram, H; Naijian, H; Bahreini, M and Poorhamdollah, M (2011). Protective effect of melatonin on sperm motility parameters on liquid storage of ram semen at 5°C. Afr. J. Biotechnol., 10: 6670-6674.
Aurich, C; Seeber, P and Muller-Schlosser, F (2007). Comparison of different extenders with defined protein composition for storage of stallion spermatozoa at 5°C. Reprod. Domest. Anim., 42: 445-448.
Bahmani, S; Eslami, M; Farrokhi-Ardabili, F; Imani, M and Batavani, RA (2023). Evaluation of chicken egg yolk plasma and low-density lipoprotein alone or enriched with ewe or cow skim milk in tris-citric acid-based diluent for cryostorage of ram semen. Biopreserv. Biobank., 21: 346-354.
Chohan, K; Griffin, J and Carrell, D (2004). Evaluation of chromatin integrity in human sperm using acridine orange staining with different fixatives and after cryopreservation. Andrologia. 36: 321-326.
Correa, JR and Zavos, PM (1994). The hypoosmotic swelling test: its employment as an assay to evaluate the functional integrity of the frozen-thawed bovine sperm membrane. Theriogenology. 42: 351-360.
Dalmazzo, A; Daniel, DSRA; Losano, JDA; Rocha, C; Sobrinho, CAB; Chinait Gurgel, JR; Monteiro Pacheco, PI; Minazaki, CK; Crusco, SE and Nichi, M (2019). Insights into soy lecithin and egg yolk-based extenders for chilling canine spermatozoa. Zygote. 1: 17-24.
Del Valle, I; Gómez-Durán, A; Holt, WV; Muiño-Blanco, T and Cebrián-Pérez, JA (2012). Soy lecithin interferes with mitochondrial function in frozen-thawed ram spermatozoa. J. Androl., 33: 717-725.
de Paz, P; Esteso, MC; Alvarez, M; Mata, M; Chamorro, CA and Anel, L (2010). Development of extender based on soybean lecithin for its application in liquid ram semen. Theriogenology. 74: 663-671.
Evans, G and Maxwell, WMC (1987). Frozen storage of semen. In: Salamon’s artificial insemination of sheep and goats. (1st Edn.), Butterworths, Wellington, Butterworth-Heinemann. PP: 122-141.
Forouzanfar, M; Sharafi, M; Hosseini, S; Ostadhosseini, S; Hajian, M; Hosseini, L; Abedi, P; Nili N; Rahmani, H and Nasr-Esfahani, M (2010). In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology. 73: 480-487.
Green, LC; Wagner, DA; Glogowski, J; Skipper, PL; Wishnok, JS and Tannenbaum, SR (1982). Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal. Biochem., 126: 131-138.
Holt, WV (2000). Basic aspects of frozen storage of semen. Anim. Reprod. Sci., 62: 3-22.
Hou, ML; Huang, SY; Lai, YK and Lee, WC (2008). Geldanamycin augments nitric oxide production and promotes capacitation in boar spermatozoa. Anim. Reprod. Sci., 104: 56-68.
Khosravi, F; Valojerdi, MR; Amanlou, M; Karimian, L and Abolhassani, F (2014). Relationship of seminal reactive nitrogen and oxygen species and total antioxidant capacity with sperm DNA fragmentation in infertile couples with normal and abnormal sperm parameters. Andrologia. 46: 17-23.
Kmenta, I; Strohmayer, C; Müller-Schlösser, F and Schäfer-Somi, S (2011). Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of cold-stored canine spermatozoa. Theriogenology. 75: 1095-1103.
MacBee, LE and Cotterill, OJ (1979). Ion-exchange chromatography and electrophoresis of egg yolk proteins. J. Food Sci., 44: 656-660.
Mata-Campuzano, M; Álvarez-Rodríguez, M; Álvarez, M; Tamayo-Canul, J; Anel, L; Paz, PD and Martínez-Pastor, F (2015). Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender. Theriogenology. 83: 520-528.
Meng, Q; Yin, J; Jin, M and Gao, H (2018). Distinct nitrite and nitric oxide physiologies in Escherichia coli and Shewanella oneidensis. Appl. Environ. Microbiol., 84: e00559-18.
Molodtsov, V; Nawarathne, IN; Scharf, NT; Kirchhoff, PD; Showalter, HD; Garcia, GA and Murakami, KS (2013). X-ray crystal structure of the Escherichia coli RNA polymerase in complex with Benzoxazinorifamycin. J. Med. Chem., 56: 4758-4763.
Moran, JM; Madejón, L; Ortega Ferrusola, C and Peňa, FJ (2008). Nitric oxide induces caspase activity in boar spermatozoa. Theriogenology. 70: 91-96.
Mortazavi, SH; Eslami, M and Farrokhi-Ardabili, F (2020). Comparison of different carrier-compounds and varying concentrations of oleic acid on freezing tolerance of ram spermatozoa in tris-citric acid-egg yolk plasma semen diluent. Anim. Reprod. Sci., 219: 106533.
Mousavi, SM; Towhidi, A; Zhandi, M; Amoabediny, G; Mohammadi-Sangcheshmeh, A; Sharafi, M and Hussaini, SMH (2019). Comparison of two different antioxidants in a nano lecithin-based extender for bull sperm cryopreservation. Anim. Reprod. Sci., 209: 106171.
Nadri, T; Towhidi, A; Zeinoaldini, S; Martínez-Pastor, F; Mousavi, M; Noei, R; Tar, M and Sangcheshmeh, AM (2019). Lecithin nanoparticles enhance the cryosurvival of caprine sperm. Theriogenology. 133: 38-44.
Najafi, A; Taheri, RA; Mehdipour, M; Martínez-Pastor; M and Rouhollahi, A (2019). Improvement of post-thawed sperm quality in broiler breeder roosters by ellagic acid loaded liposomes. Poult. Sci., 98: 440-446.
Oztezcan, S; Turkoglu, UM; Kervancioglu, E; Kocak, T; Kocak-Toker, N and Aykac-Toker, G (1999). In vitro effects of peroxynitrite on human spermatozoa. Andrologia. 31: 195-198.
Ricker, JV; Linfor, JJ; Delfino, WJ; Kysar, P; Scholtz, EL; Tablin, F; Crowe, JH; Ball, BA and Meyers, SA (2006). Equine sperm membrane phase behavior: the effects of lipid-based cryoprotectants. Biol. Reprod., 74: 359-365.
Röpke, T; Oldenhof, H; Leiding, C; Sieme, H; Bollwein, H and Wolkers, W (2011). Liposomes for cryopreservation of bovine sperm. Theriogenology. 76: 1465-1472.
Salamon, S and Maxwell, WMC (1995). Frozen storage of ram semen. I. Processing, freezing, thawing and fertility after cervical insemination. Anim. Reprod. Sci., 37: 185-249.
Santiso, R; Tamayo, M; Gosálvez, J; Johnston, S; Mari, A; Fernández, C; Losada, C and Fernández, JL (2012). DNA fragmentation dynamics allows the assessment of cryptic sperm damage in human: evaluation of exposure to ionizing radiation, hyperthermia, acidic pH and nitric oxide. Mutat. Res-/Fund. Mol. Mech. Mut., 734: 41-49.
Serrano, R; Garrido, N; Céspedes, JA; González-Fernández, L; García-Marín, LJ and Bragado, MJ (2020). Molecular mechanisms involved in the impairment of boar sperm motility by peroxynitrite-induced nitrosative stress. Int. J. Mol. Sci., 21: 1208.
Singh, AK; Kumar, A; Honparkhe, M; Kaur, S and Brar, PS (2017). Comparison of in vitro and in vivo fertilizing potential of buffalo bull semen frozen in egg yolk-, soya bean lecithin- and liposome-based extenders. Reprod. Domest. Anim., 53: 195-202.
Stern, ST; Potter, TM and Neun, BW (2010). Hep G2 hepatocyte lipid peroxidation assay. NCL Method GTA-4. Version 1.1. National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]. Bethesda (MD): National Cancer Institute (US).
doi: 10.17917/H65D-SJ70.
Sun, LW; Fan, WH; Wu, CF; Zhang, SS; Dai, JJ and Zhang, DF (2020). Effect of substituting different concentrations of soybean lecithin and egg yolk in tris-based extender on goat semen cryopreservation. Cryobiology. 92: 146-150.
Sun, L; He, M; Wu, C; Zhang, S; Dai, J and Zhang, D (2021). Beneficial influence of soybean lecithin nanoparticles on rooster frozen-thawed semen quality and fertility. Animals. 11: 1769.
Uribe, P; Boguen, R; Treulen, F; Sánchez, R and Villegas, JV (2015). Peroxynitrite-mediated nitrosative stress decreases motility and mitochondrial membrane potential in human spermatozoa. Mol. Hum. Reprod., 21: 237-243.
Vignini, A; Nanetti, L; Buldreghini, E; Moroni, C; Ricciardo-Lamonica, G; Mantero, F; Boscaro, M; Mazzanti, L and Balercia, G (2006). The production of peroxynitrite by human spermatozoa may affect sperm motility through the formation of protein nitrotyrosine. Fertil. Steril., 85: 947-953.
Wojtusik, J; Stoops, MA and Roth, TL (2018). Comparison of soy lecithin, coconut water, and coconut milk as substitutes for egg-yolk in semen cryodiluent for black rhinoceros (Diceros bicornis) and Indian rhinoceros (Rhinoceros unicornis). Theriogenology. 121: 72-77.
Yeste, M (2016). Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs. Theriogenology. 85: 47-64.
Zhang, SS; Hu, JH; Li, QW; Jiang, ZL and Zhang, XY (2009). The cryoprotective effects of soybean lecithin on boar spermatozoa quality. Afr. J. Biotechnol., 8: 6476-6480.

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