1. Effects of slow freezing and vitrification of human semen on post-thaw semen quality and miRNA expressionRebeka Podgrajšek, Luka Bolha, Tjaša Pungert, Jože Pižem, Katerina Jazbec Gradišar, Elvira Maličev, Martin Štimpfel, 2024, izvirni znanstveni članek Povzetek: Semen cryopreservation has played an important role in medically assisted reproduction for decades. In addition to preserving male fertility, it is sometimes used for overcoming logistical issues. Despite its proven clinical usability and safety, there is a lack of knowledge of how it affects spermatozoa at the molecular level, especially in terms of non-coding RNAs. Therefore, we conducted this study, where we compared slow freezing and vitrification of good- and poor-quality human semen samples by analyzing conventional sperm quality parameters, performing functional tests and analyzing the expression of miRNAs. The results revealed that cryopreservation of normozoospermic samples does not alter the maturity of spermatozoa (protamine staining, hyaluronan binding), although cryopreservation can increase sperm DNA fragmentation and lower motility. On a molecular level, we revealed that in both types of cryopreservation, miRNAs from spermatozoa are significantly overexpressed compared to those in the native semen of normozoospermic patients, but in oligozoospermic samples, this effect is observed only after vitrification. Moreover, we show that expression of selected miRNAs is mostly overexpressed in native oligozoospermic samples compared to normozoospermic samples. Conversely, when vitrified normozoospermic and oligozoospermic samples were compared, we determined that only miR-99b-5p was significantly overexpressed in oligozoospermic sperm samples, and when comparing slow freezing, only miR-15b-5p and miR-34b3p were significantly under-expressed in oligozoospermic sperm samples. Therefore, our results imply that cryopreservation of normozoospermic sperm samples can modulate miRNA expression profiles in spermatozoa to become comparable to those in oligozoospermic samples.
Ključne besede: semen, cryopreservation, vitrification, slow freezing, spermatozoa, microRNA, assisted reproduction, infertility
Objavljeno v DiRROS: 10.06.2026; Ogledov: 121; Prenosov: 112
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3. Molecular and cellular mechanisms of the therapeutic effect of mesenchymal stem cells and extracellular vesicles in corneal regenerationNina Kobal, Miha Marzidovšek, Petra Schollmayer, Elvira Maličev, Marko Hawlina, Zala Lužnik Marzidovšek, 2024, pregledni znanstveni članek Povzetek: The cornea is a vital component of the visual system, and its integrity is crucial for optimal vision. Damage to the cornea resulting from trauma, infection, or disease can lead to blindness. Corneal regeneration using mesenchymal stem cells (MSCs) and MSC-derived extracellular vesicles (MSC-EVs) offers a promising alternative to corneal transplantation. MSCs are multipotent stromal cells that can differentiate into various cell types, including corneal cells. They can also secrete a variety of anti-inflammatory cytokines and several growth factors, promoting wound healing and tissue reconstruction. This review summarizes the current understanding of the molecular and cellular mechanisms by which MSCs and MSC-EVs contribute to corneal regeneration. It discusses the potential of MSCs and MSC-EV for treating various corneal diseases, including corneal epithelial defects, dry eye disease, and keratoconus. The review also highlights finalized human clinical trials investigating the safety and efficacy of MSC-based therapy in corneal regeneration. The therapeutic potential of MSCs and MSC-EVs for corneal regeneration is promising; however, further research is needed to optimize their clinical application.
Ključne besede: cell-based therapy, corneal epithelium, corneal regeneration, corneal stroma, dry eye disease, extracellular vesicles, immune and inflammatory modulation, keratoconus, limbal stem cell deficiency, mesenchymal stem cells
Objavljeno v DiRROS: 08.12.2025; Ogledov: 484; Prenosov: 367
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4. Extracellular vesicle-derived bioactive molecules for corneal and ocular surface regenerationAna Kolenc, Živa Dimnik, Miha Marzidovšek, Petra Schollmayer, Marko Hawlina, Elvira Maličev, Zala Lužnik Marzidovšek, 2025, pregledni znanstveni članek Povzetek: Cell-based therapies emerge as potential treatment options for various debilitating diseases. Preclinical research and clinical studies involving cells increased exponentially in the past decade. In addition to cell-based approaches, the use of extracellular vesicles (EVs), which are released by nearly all cell types, emerged as a promising cell-free alternative. Those approaches are also being explored in the field of ophthalmology. Several clinical trials involving EVs are underway to develop potential treatments for advanced ocular surface diseases, including corneal disorders, injuries, and dry eye disease. The cargo carried by EVs has been shown to include a diverse array of functional molecules such as transcription factors, cytokines, growth factors, mRNA, tRNA, rRNA, miRNA, and fragments of dsDNA. While the molecular composition of EVs is already well characterised, the specific activity of these molecules upon delivery to recipient cells remains poorly understood. In this review, we summarise recent studies investigating the bioactive molecules within EVs shown to influence or modulate cellular activity on the ocular surface. Among these, various miRNAs have most commonly been identified as therapeutic agents targeting distinct molecular pathways. The EVs studied were predominantly derived from various mesenchymal stem cells.
Ključne besede: extracellular vesicles, bioactive molecules, cornea, dry eye disease, miRNA, mesenchymal stem cells, ophthalmology
Objavljeno v DiRROS: 24.11.2025; Ogledov: 1070; Prenosov: 374
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