Document Type : Original Article
Authors
1
PhD Student, Reproductive Physiology Lab, Department of Physiology, University of Kurdistan, Sanandaj, Kurdistan, Iran
2
Laboratory of Reproductive Physiology, University of Kurdistan, Kurdistan, Sanandaj, Iran
3
Professor, Laboratory of Physiopathology and Reproduction Research, UCLouvain, Brussels, Belgium
4
Professor, Department of Anatomy, Tehran University of Medical Sciences, Tehran, Iran
10.22038/ijogi.2025.89724.6496
Abstract
Background:
Although ovarian tissue cryopreservation and thawing play an important role in fertility preservation, they often lead to structural damage and a reduction in follicle survival. These injuries are mainly caused by osmotic and mechanical stresses and alterations in the tissue microenvironment. Alginate hydrogel, due to its biocompatibility and ability to control mechanical properties, has been proposed as a potential protective system for ovarian tissue encapsulation.
Methods:
Alginate hydrogel was formulated using alginate, calcium carbonate (CaCO₃), and glucono-δ-lactone (GDL) and optimized through the response surface methodology (RSM) to achieve storage modulus and gelation time comparable to natural ovarian tissue. Ovarian tissue fragments were encapsulated in the hydrogel, cryopreserved using a slow-freezing protocol, and cultured for 24 hours after thawing. Histological evaluations included counting primordial and primary follicles, as well as assessing stromal cells and extracellular matrix integrity. Data were analyzed using GraphPad Prism version 9, and comparisons were performed with Duncan’s test. A significance level of p<0.05 was considered.
Results:
The density of primordial and primary follicles in the encapsulated group was significantly higher than in the unprotected frozen group (p<0.05). Stromal cell integrity were also better preserved in the hydrogel group (p<0.001). The hydrogel exhibited a storage modulus of approximately 1750 Pa and a gelation time of around 30 minutes, closely mimicking the mechanical properties of natural ovarian tissue.
Conclusion:
Encapsulation of ovarian tissue within alginate hydrogel effectively reduces cryo-induced damage and improves follicular survival and stromal preservation. This approach represents a promising and innovative strategy for fertility preservation.
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