This comprehensive review critically examines the application of proteomics in understanding sperm cryoinjury mechanisms in livestock animals, in the context of the widespread use of semen cryopreservation for genetic conservation. Despite its global adoption, cryopreservation often detrimentally affects sperm quality and fertility due to cryoinjuries. These injuries primarily arise from ice crystal formation, osmotic shifts, oxidative stress, and the reorganization of membrane proteins and lipids during freezing and thawing, leading to premature capacitation-like changes. Moreover, the cryopreservation process induces proteome remodeling in mammalian sperm. Although there have been technological advances in semen cryopreservation, the precise mechanisms of mammalian sperm cryoinjury remain elusive. This review offers an in-depth exploration of how recent advancements in proteomic technologies have enabled a detailed investigation into these molecular disruptions. It presents an analysis of protein-level alterations post-thaw and their impact on sperm viability and functionality. Additionally, it discusses the role of proteomics in refining cryopreservation techniques to mitigate cryoinjury and enhance reproductive outcomes in livestock. This work synthesizes current knowledge, highlights gaps, and suggests directions for future research in animal reproductive science and biotechnology.

The Cryopreservation of spermatozoa ensures preserving fertility potential after some medical treatments such as chemotherapy and radiotherapy in cancer patients. However, many spermatozoa encounter serious damages, and their motility and viability decrease considerably after thawing. The excessive production of reactive oxygen species is one of the major causes of these damages. The supplementation of cryopreservation media with vitamins, which are well-known antioxidants, can reduce cryopreservation-induced damages. In this systematic review, we aimed to evaluate the cryoprotective effect of various vitamins on the quality of cryopreserved-thawed human spermatozoa. Two researchers searched PubMed, ISI, and Scopus databases up to March 2020. All original articles using vitamins in human spermatozoa cryopreservation media were included. We used a standardized form to extract sample size and to determine sample quality, the type and dose of vitamins, and the cryopreservation methods and their effects. We performed a meta-analysis on studies with available data (Mean + SD in cryoprotectant and cryoprotectant + cryoprotectant groups). We also performed a test of between-study heterogeneity, subgroup analysis, and meta-regression. Out of 258 studies, 16 articles were included for the analysis. Our meta-analysis revealed that using vitamins in cryopreservation media could increase motility by 4.60% (95% CI 6.16, 3.05; P = 0.0001), viability by 5.71% (95% CI 9.71, 1.72; P = 0.0001), and DNA integrity by 10.20% (95% CI 12.98, 7.42; P = 0.0001) in cryopreserved-thawed spermatozoa. We found a significant correlation between using vitamins and improved spermatozoa quality; the sperm motility and viability were improved and DNA fragmentation was reduced after thawing by vitamins. However, we could not emphasize on any type or dose of vitamins but we conclude that the anti-oxidative function of vitamins is the main reason for these benefits.