The Influence of Platelet-Derived Growth Factors on the Proliferation of Germinal Epithelium After Local Irradiation with Electrons.

Background: At present, the damage of male reproductive function caused by electron irradiation, as well as the development of methods for its correction are the relevant topics for further research. In fact, the effect of leukocyte-poor platelet-rich plasma (LP-PRP) growth factors are poorly investigated on different aspects of male fertility. Methods: In this study, Wistar rats were divided into four groups; I) control which were injected with saline; II and III) groups (n=65) whose testes were locally irradiated with electrons to a dose of 2 Gy (linear accelerator "NOVAC-11"); III) the group (n=30) which received LP-PRP for 11 weeks after irradiation; and IV) animals (n=30) which received only LP-PRP (conditional control). The testes were studied by histological, immunohistochemical (IHC), western blotting, and TUNEL methods using Ki-67, Bcl-2, and p53. Comparison between groups was performed and p<0.01 was set as the level of significance. Results: The results showed a decrease in the expression levels of Ki-67 and Bcl-2 besides an increase in p53-positive cells by the end of the experiment (p<0.01). After injection of LP-PRP, a gradual restoration of the proliferative activity of gametes was noted, which was confirmed by an increase in the proportion of Ki-67- and Bcl-2-positive germ cells (46.4±2.3, p<0.01 and 23.5±1.1, respectively, p<0.01). Conclusion: Ki-67 expression and TUNEL analysis in the testes revealed a modulation of the proliferative-apoptotic balance towards apoptosis of germ cells after 2 Gy local electron irradiation. A tendency to restore the proliferative-apoptotic balance was noted after LP-PRP injections as indicated by increase in Ki-67-positive germ cells.

Morphological Characteristics of the Lacrimal Apparatus in its Obstruction of Various Genesis.

Secondary acquired lacrimal duct obstruction (SALDO) is one of the complications of radioiodine therapy. The material was obtained during endoscopic dacryocystorhinostomy with revision of Hasner's valve in patients with PANDO (n = 7) in the distal segments of the nasolacrimal duct and in patients with SALDO (n = 7) after radioactive iodine therapy. The material was stained with hemotoxylin and eosin, alcyan blue, and by Masson method. Morphological and morphometric analyses were performed in semi-automatic mode. The results of histochemical staining of sections were translated into points taking into account the area and optical density (chromogenicity). The differences were considered significant at p < 0.05. It was shown that the nasolacrimal duct sclerosis is significantly lower (p = 0.029) in patients with SALDO than in patients with PANDO while fibrosis in the lacrimal sac is the same in patients of the compared groups.

Immunohistochemical Analysis of Spermatogenesis in Patients with SARS-CoV-2 Invasion in Different Age Groups.

Based on studies that focused on the effect of SARS-CoV-2 on human tissues, not only pulmonary invasion was revealed, but also impaired testicular function. Thus, the study of the mechanisms of influence of SARS-CoV-2 on spermatogenesis is still relevant. Of particular interest is the study of pathomorphological changes in men of different age groups. The purpose of this study was to evaluate immunohistochemical changes in spermatogenesis during SARS-CoV-2 invasion in different age groups. In our study, for the first time, a cohort of COVID-19-positive patients of different age groups was collected, and the following were conducted--confocal microscopy of the testicles and immunohistochemical evaluation of spermatogenesis disorders in SARS-CoV-2 invasion with antibodies to the spike protein, the nucleocapsid protein of the SARS-CoV-2 virus, and angiotensin convertase type 2. An IHC study and confocal microscopy of testicular autopsies from COVID-19-positive patients revealed an increase in the number of S-protein- and nucleocapsid-positively stained spermatogenic cells, which indicates SARS-CoV-2 invasion into them. A correlation was found between the number of ACE2-positive germ cells and the degree of hypospermatogenesis, and in the group of patients with confirmed coronavirus infection older than 45 years, the decrease in spermatogenic function was more pronounced than in the cohort of young people. Thus, our study found a decrease in both spermatogenic and endocrine (Leydig cells) testicular functions in patients with COVID-19 infection. In the elderly, these changes were significantly higher than in the group of young patients.

Properties and Printability of the Synthesized Hydrogel Based on GelMA.

Gelatin methacryloyl (GelMA) has recently attracted increasing attention. Unlike other hydrogels, it allows for the adjustment of the mechanical properties using such factors as degree of functionalization, concentration, and photocrosslinking parameters. In this study, GelMA with a high degree of substitution (82.75 ± 7.09%) was synthesized, and its suitability for extrusion printing, cytocompatibility, and biocompatibility was studied. Satisfactory printing quality was demonstrated with the 15% concentration hydrogel. The high degree of functionalization led to a decrease in the ability of human adipose-derived stem cells (ADSCs) to adhere to the GelMA surface. During the first 3 days after sowing, proliferation was observed. Degradation in animals after subcutaneous implantation was slowed down.

On the Pathomorphological Pattern of the Efficiency of Photodynamic Therapy of Murine Melanoma B16 Using a New Photosensitizer Based on Chlorin e6 Conjugate with a Prostate-Specific Membrane Antigen.

Photodynamic therapy (PDT) is an effective treatment for a number of solid malignancies. In this work, the antitumor efficacy of photodynamic therapy for murine B16 melanoma with intravenous administration of a new photosensitizer (PS) based on the chlorin e6 conjugate with a prostate-specific membrane antigen (PSMA) was studied in vivo. We have previously published the data obtained in the first part of the study: the dynamics of PS accumulation in the tumor and surrounding tissues and the antitumor efficacy of the photodynamic therapy, which was evaluated by the regression parameters and morphological characteristics of the tumors-including by the complete regression of the tumors, the absolute growth rate of the tumors among the mice with continued tumor growth, and an increase in life expectancy compared to the control. The criterion for a complete cure was the absence of signs of tumor recurrence within 90 days after therapy. The conducted studies demonstrated the high efficiency of the new photosensitizer for the photodynamic therapy of B16 melanoma. This article presents a continuation of this work, including histological studies of the zones exposed to laser irradiation on the 21st day after treatment and an assessment of the therapeutic potential of photodynamic therapy for the destruction of tumor cells. Pathological studies in the zones of photodynamic exposure revealed that the effectiveness of the PDT depended on the PS dose and the laser irradiation parameters.

Cartilage Formation In Vivo Using High Concentration Collagen-Based Bioink with MSC and Decellularized ECM Granules.

The aim of this study was to verify the applicability of high-concentration collagen-based bioink with MSC (ADSC) and decellularized ECM granules for the formation of cartilage tissue de novo after subcutaneous implantation of the scaffolds in rats. The printability of the bioink (4% collagen, 2.5% decellularized ECM granules, derived via 280 µm sieve) was shown. Three collagen-based compositions were studied: (1) with ECM; (2) with MSC; (3) with ECM and MSC. It has been established that decellularized ECM granules are able to stimulate chondrogenesis both in cell-free and MSC-laden scaffolds. Undesirable effects have been identified: bone formation as well as cartilage formation outside of the scaffold area. The key perspectives and limitations of ECM granules (powder) application have been discussed.

An experimental approach to comprehend the influence of platelet rich growth factors on spermatogenesis.

MATERIALS AND METHODS: Experimental animals (n = 135) were divided into 5 groups: I - control (n = 10); II - 2IR (n = 35; 2 Gy); III - 2IR + LP-PRP + IGF-1 (n = 30); IV - 2IR + LP-PRP (n = 30); V - LP-PRP (n = 30). RESULTS: Electron irradiation reduces the number of germ cells in comparison with the control group. After injection of LP-PRP + rhIGF-1 significantly increased the number of germ cells, Sertoli and Leydig cells, the height of germinal epithelium, area and diameter of seminiferous tubules. CONCLUSION: LP-PRP + rhIGF-1 has a normalizing effect on structural and functional disorders of the testis caused by electron irradiation.

Bioprinting of Cartilage with Bioink Based on High-Concentration Collagen and Chondrocytes.

The study was aimed at the applicability of a bioink based on 4% collagen and chondrocytes for de novo cartilage formation. Extrusion-based bioprinting was used for the biofabrication. The printing parameters were tuned to obtain stable material flow. In vivo data proved the ability of the tested bioink to form a cartilage within five to six weeks after the subcutaneous scaffold implantation. Certain areas of cartilage formation were detected as early as in one week. The resulting cartilage tissue had a distinctive structure with groups of isogenic cells as well as a high content of glycosaminoglycans and type II collagen.

Inhibin B in seminiferous tubules of human testes in normal spermatogenesis and in idiopathic infertility.

Inhibin B is one of the most significant serum markers of spermatogenesis, but its testicular expression has been poorly studied. Inhibin B sensitivity as well as the ability to reflect the level and condition of spermatogenesis and forecast certain changes in reproductive homeostasis in males is still a subject of active discussion. The purpose of this study is to examine the level of expression of inhibin B in cells of human seminiferous tubules in normal and in pathological spermatogenesis (idiopathic infertility) by revealing the proportion of immunostaining cells for inhibin B. The research conducted included analysis of testicular tissue samples taken from 82 males diagnosed with infertility and nonobstructive azoospermia. The influence of inhibin on the germ cells of men aged 22-35 been analyzed using the immunohistochemical method. According to the obtained results, high expression of inhibin can be detected both in Sertoli (98.0 ± 2.66) and Leydig (94.0 ± 1.55) cells in patients suffering from focal spermatogenesis disorders (mixed atrophy), in comparison with the men in the control group (65.9 ± 0.44 and 12.0 ± 0.44, respectively). The level of inhibin expression in the cytoplasm of spermatogonia was 4.0 ± 0.22 in control group of fertile men, while it was significantly increased in patients with Sertoli cell-only syndrome with focal spermatogenesis (45.0 ± 0.44). In the case of severe lesion to the seminiferous tubules (e.g., tubular hyalinization), the lowest level of inhibin expression in Sertoli cells is detected, whereas the immunostaining in Leydig cells showed only slight changes. Further histological research of Sertoli cells and inhibin B expression is necessary because, according to our data, the degree of inhibin B expression may be a useful marker of Sertoli cells function which can lead to new findings in the concept of local reproductive homeostasis in testis that may be impaired in some forms of idiopathic infertility in males. Abbreviations: ß-TGF: ß-transforming growth factor family; GCA: germ cell abnormalities/atypia; JS: Johnsen score; FSH: follicle stimulating hormone; TESE: testicular sperm extraction; LH: luteinizing hormone; F-Testo: free testosterone; ELISA: the enzyme-linked immunosorbent assay; CV: coefficient of variance; DR: range of definitions; AZF: azoospermia factorI; HC: immunohistochemical; HIER: heat induction of epitope retrieval; H&E: hematoxylin and eosin.