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. 2025 Aug 12;61(8):1450.
doi: 10.3390/medicina61081450.

Non-Obstructive Azoospermia: Influence of PRP on Proliferation, Apoptosis, and Growth Factors of Male Germ Cells

Affiliations

Non-Obstructive Azoospermia: Influence of PRP on Proliferation, Apoptosis, and Growth Factors of Male Germ Cells

Grigory Demyashkin et al. Medicina (Kaunas). .

Abstract

Background and Objectives: Currently, infertility is one of the major problems affecting up to 12% of couples worldwide, with more than a quarter of cases being male-related. It is assumed that Leukocyte-poor platelet-rich plasma (LP-PRP) can improve the function of germ cells and serve as a regenerative substrate as a source of biologically active substances that play an important role in the process of spermatogenesis in infertile men. We aimed to evaluate the proliferation, apoptosis, and growth factors of germ cells after the administration of LP-PRP in patients with non-obstructive azoospermia. Materials and Methods: The study used archival material (paraffin blocks of testicular biopsies) of patients with non-obstructive azoospermia aged 21-34 years (n = 41; associated diagnosis: varicocele). We confirm that no interventions or biopsies were performed as part of the study itself. They were injected bilaterally into the spermatic cord and in the region of the lower pole of the testis under ultrasound control were injected with PRP once a week for 6 weeks. Biopsies were immunohistochemical reactions with antibodies to Ki-67, Bcl-2, caspase 3 and p53, IGF-1, TGF-β, and VEGF-A. Results: Immunohistochemical study of testicular biopsies after LP-PRP injection revealed an increase in the number of cells stained for proliferation proteins (Ki-67) and anti-apoptosis (Bcl-2), IGF-1, TGF-β, VEGF-A; decrease caspase-3- and p53-positive cells. Conclusions: In LP-PRP, platelet α-granule growth factors, which are key regulators of the cell cycle of germ cells, demonstrate restoration of the proliferative-apoptotic balance, confirmed by the expression levels of Ki-67, Bcl-2, caspase 3, and p53 in patients with non-obstructive azoospermia. In human testicular biopsies, the administration of LP-PRP led to an exponential release of numerous growth factors from platelet α-granules, which, based on their regenerative properties, improved the morphological and immunohistochemical picture of the germinal epithelium in non-obstructive azoospermia.

Keywords: PRP; growth factors; non-obstructive azoospermia; spermatogenesis.

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Conflict of interest statement

The authors declare no competing financial interests. The authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

Figures

Figure 1
Figure 1
(Left)—Technique of PRP injections for male infertility: A—intracanalicular; B—intratesticular. (Center)—PRP injection technique for male infertility (intracanalicular under ultrasound control). (Right)—Movement of the PRP to the epididymis through 8 min (green triangle—epididymis; green arrow—intracanalicular deposition of PRP).
Figure 2
Figure 2
Seminiferous tubules of testicular biopsy before and after LP-PRP administration. The arrow indicates Leydig cell hyperplasia in the interstitial tissue. Staining: hematoxylin and eosin; magn.: ×400; bar: 25 μm.
Figure 3
Figure 3
Morphological analysis of testicular biopsies: A—seminiferous tubule diameter (µm); B—germinal epithelium height (µm); * p < 0.05—significant difference between ‘Before’ and ‘After’ treatment groups.
Figure 4
Figure 4
Morphological analysis of testicular biopsies: A—cross-sectional area of the seminiferous tubule (µm2); B—interstitial tissue area (µm2); * p < 0.05—significant difference between ‘Before’ and ‘After’ treatment groups.
Figure 5
Figure 5
Morphological analysis of testicular biopsies: (A)—mean number of germ cells in the seminiferous tubule; (B)—number of Leydig cells per 1 mm2; * p < 0.05—significant difference between ‘Before’ and ‘After’ treatment groups.
Figure 6
Figure 6
Seminiferous tubules of testicular biopsies before and after LP-PRP injection. Immunohistochemical reactions with antibodies to Ki-67, Bcl-2, caspase 3, p53 (finishing with hematoxylin); magn. ×400. Bar—25 μm.
Figure 7
Figure 7
Seminal tubules of testicular biopsy specimens from patients with non-obstructive azoospermia exhibit twisted morphology before and after LP-PRP administration. The immunohistochemical reactions’ positive results for anti-IGF-1, anti-TGF-β, and anti-VEGF-A. Magn. ×400. Bar—25 μm.
Figure 8
Figure 8
The positive effects of PRP components on spermatogenesis in patients with non-obstructive azoospermia; arrows pointing up—increase, arrows pointing down—decrease.

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