Laboratory of Nanotechnology and Nanomedicine (LNMED), Institute of Technology and Research (ITP), Av. Murilo Dantas 300, Aracaju, 49010-390, Brazil.
Postgraduate Program in Industrial Biotechnology (PBI), Tiradentes University (UNIT), Av. Murilo Dantas 300, Aracaju, 49032-490, Brazil.
Acta Vet Scand. 2020 Jun 18;62(1):31. doi: 10.1186/s13028-020-00530-6.
Cryopreservation of stallion spermatozoa tends to cause plasma membrane damage due to the low ratio of cholesterol to phospholipids. Gums have been suggested as an alternative cryoprotectant to glycerol for stallion spermatozoa. Therefore, the present experiment was designed to verify whether the effect of addition of cashew gum (CG), or nanoparticles (NP) containing CG, to the extender before cooling on sperm quality in stallion semen. Ejaculates from 6 stallions were extended and split between six treatment groups (control, a-tocopherol [TOC], CG1, CG0.5, NP1 and NP0.5), stored in cryotubes at 4 °C.
Aliquots were analysed by computer-assisted sperm motility analysis on the day of collection, and after 24 h and 48 h of cold storage. After 48 h, the total motility with NP1 (78.53 + 6.31%) was similar to control 85.79 + 6.31% at 0 h. The same pattern was observed for progressive motility. Membrane integrity assessed by flow cytometer was similar between control, TOC and G1 at all storage times. The DNA fragmentation in the control group increased at all time points, whereas chromatin integrity was maintained after 24 h in TOC and NP0.5 compared to 0 h. There was no increase in the proportion of live spermatozoa producing hydrogen peroxide, but there was a tendency for an increased proportion of spermatozoa in the live superoxide category in CG1 after 24 h cooled storage.
The addition of CG or CG-derived NP to extender for stallion semen was not harmful to the sperm cells.
由于胆固醇与磷脂的比例低，精子的冷冻保存往往会导致质膜受损。树胶已被提议作为甘油的替代冷冻保护剂用于马精液的精子。因此，本实验旨在验证在冷却前将腰果胶（CG）或含 CG 的纳米颗粒（NP）添加到稀释液中是否会对种马精液中的精子质量产生影响。来自 6 匹种马的精液被扩展并分为六个处理组（对照组、生育酚 [TOC]、CG1、CG0.5、NP1 和 NP0.5），在 4°C 的冷冻管中储存。
在采集当天、24 小时和 48 小时冷储存后，通过计算机辅助精子运动分析对样本进行分析。48 小时后，NP1 的总活力（78.53 + 6.31%）与 0 小时的对照组 85.79 + 6.31%相似。同样的模式也适用于渐进性活力。通过流式细胞仪评估的膜完整性在所有储存时间内均与对照组、TOC 和 G1 相似。对照组的 DNA 片段化在所有时间点都增加，而 TOC 和 NP0.5 组在 24 小时后保持了染色质完整性，与 0 小时相比。产生过氧化氢的活精子比例没有增加，但 CG1 组在 24 小时冷储存后活超氧化物类别中的精子比例有增加的趋势。
向种马精液的稀释液中添加 CG 或 CG 衍生的 NP 对精子细胞没有危害。
