Platelet-Rich Plasma in Vitrification; is it Helpful or Harmful?
Türker Çavuşoğlu1*,
Aylin Gökhan2, Canberk Tomruk3, Cansın Şirin2,
Kubilay Doğan Kılıç2 and Gürkan Yiğittürk4
1Department
of Histology and Embryology, Faculty of Medicine, Izmir Bakircay
University, Izmir, Turkey; 2Department of Histology and
Embryology, Faculty of Medicine, Ege University, Izmir, Turkey;
3Department of Histology and Embryology, Republic of Turkey
Ministry of Health Samsun Education and Research Hospital, Samsun,
Turkey; 4Department of Histology and Embryology, Faculty
of Medicine, Mugla Sitki Kocman University, Izmir, Turkey
*Corresponding author:
turker.cavusoglu@bakircay.edu.tr
Abstract
Human and animal studies on cryoprotectants and freezing
solutions are still needed to establish a simple yet reliable protocol and
increase the success of cryopreservation. The main aim of this study was to
evaluate the short- and long-term effects of platelet-rich plasma, a well-known
antioxidant substance due to its contents including bioactive molecules and
growth factors, on whole ovarian tissue cryopreservation. Fresh tissues (control
group, G1) were subjected to histological tissue processing without any
treatment. Ovaries treated with platelet-rich plasma (PRP)-supplemented
vitrification solution were subjected to tissue processing without cryostorage
group 2 (G2) or following six months of cryostorage group 3 (G3). Steps in G2
and G3 were also performed for group 4 (G4) and group 5 (G5), respectively,
except that the vitrification solution was supplemented with fetal bovine serum.
PRP was activated with calcium chloride (CaCl2) after double
centrifugation. Ethylene glycol, dimethyl sulfoxide, and sucrose were used as
cryoprotective agents in all groups. Histomorphological changes were evaluated
with the semi-quantitative histochemical-scoring algorithm. Apoptotic and
antiapoptotic effects and intercellular connections were evaluated with
immunohistochemical staining of Bax, Bcl-2, Caspase-3 (C3), Connexin-43 (Cx-43),
and TUNEL analysis. Cryopreservation with PRP-supplementation (G3) significantly
increased tissue degeneration (p<0.05). There was an increase in the number of
degenerated both primary and secondary follicles (p<0.05), and an increase in
the immune expression of Bax, C3 and Cx-43 and TUNEL assay in G3 was observed
compared to other groups (p<0.05). Since the morphology of primordial follicles
was more preserved than other follicles in all groups, primordial follicles were
not included in the follicle count. Our study suggested that cryopreservation
with PRP-supplemented vitrification solution caused excessive damage to rat
ovaries. We assumed that CaCl2 might have further provoked this
cellular damage.
To Cite This Article:
Çavuşoğlu T, Gökhan A,
Tomruk C, Şirin C, Kılıç KD, Yiğittürk G,
2023. Platelet-rich plasma in
vitrification; is it helpful or harmful?.
Pak Vet J, 43(3): 500-506. http://dx.doi.org/10.29261/pakvetj/2023.077