Protective effect of platelet-rich plasma against structural and functional changes of the adult rat testis in carbimazole-induced hypothyroidism.

Objective: Hypothyroidism (HT) influences spermatogenesis and is associated with male infertility. Platelet-rich plasma (PRP), a biological product rich in growth factors, promotes tissue repair. In this study, the likely protective effects of PRP on testicular tissue damage in carbimazole (CBZ)-induced HT were evaluated. Methods: Forty male rats were divided into four groups. HT was induced by administering CBZ (1.35 mg/kg orally, for 45 days). Two doses of PRP (40 µL each, locally injected into the testis on days 15 and 30) were also given. After 45 days, blood samples were taken from the heart to measure triiodothyronine (T3), thyroxine (T4), and testosterone levels, and semen analysis was performed. For stereological assessment, the left testis was removed, fixed, embedded, sectioned, and stained with hematoxylin and eosin. The right testis was excised to evaluate antioxidant levels. Results: CBZ was demonstrated to induce HT, characterized by significant reductions in T3 and T4. HT was associated with decreased testicular weight, impaired sperm parameters, reduced testosterone concentration, diminished antioxidant activity, reduced volumes of testicular components, and lower total numbers of testicular cells of various types. When HT samples were treated with PRP, improvement was observed for all of these changes. This protective effect could be attributed to the growth factors present in PRP. Conclusion: PRP appears to prevent the structural changes in the testes and the deterioration in sperm quality caused by CBZ-induced HT. This protective effect is likely due to mitigation of oxidative damage and elevation of testosterone levels.

Bone repair potential of collagen-poly(3-hydroxybutyrate)-carbon nanotubes scaffold loaded with mesenchymal stem cells for the reconstruction of critical-sized mandibular defects.

The poor structural stability of collagen (COL) upon hydration poses a significant challenge in tissue engineering (TE). To overcome this limitation, the incorporation of hydrophobic polymers such as poly(3-hydroxybutyrate) (PHB), and nanomaterials such as carbon nanotubes (CNTs) has been explored. In this study, we investigated the physical, chemical, and biological characteristics of COL-based scaffolds modified with PHB and CNTs for bone tissue engineering (BTE) applications. The tensile strength analysis revealed a substantial improvement in the ultimate tensile strength with the addition of 10 % PHB and 4 % CNTs. Scanning electron microscopy (SEM) images depicted a denser and more compact structure resulting from the presence of PHB and CNTs, enhancing the scaffold's mechanical properties. Fourier-transform infrared spectroscopy (FTIR) confirmed the successful incorporation of PHB and CNTs into the composite scaffold, maintaining the chemical integrity of COL. Stereological studies also conducted in a rat model with induced critical-sized bone defects in the mandibular bone further emphasize the substantial increase in bone formation and reduction in defect volume achieved by the scaffold loaded with stem cells. These findings underscore the promising approach to enhance bone healing, using COL-based scaffolds loaded with stem cells, and the favorable results obtained in this study can contribute to the advancement of BTE strategies.

Bilirubin improves renal function by reversing the endoplasmic reticulum stress and inflammation in the kidneys of type 2 diabetic rats fed high-fat diet.

Diabetic kidney disease (DKD), as a chronic diabetes-induced complication, is considered the most frequent leading cause of end-stage renal disease (ESRD). Regarding the observed protective effects of bilirubin, as a potential endogenous antioxidant/anti-inflammatory compound, against DKD progression, we planned to evaluate the effects of bilirubin administration on endoplasmic reticulum (ER) stress and inflammation in type 2 diabetic (T2D) rats fed high-fat diet (HFD). In this regard, thirty 8-week adult male Sprague Dawley rats were divided into five groups (n = 6). T2D and obesity were induced by streptozotocin (STZ) (35 mg/kg) and HFD (700 kcal/day), respectively. Bilirubin treatment was carried out for 6- and 14-week intervals (10 mg/kg/day), intraperitoneally. Then, the expression levels of ER stress-related genes (i.e. binding immunoglobulin protein (Bip), C/EBP homologous protein (Chop), and spliced x-box-binding protein 1 (sXbp1), as well as nuclear factor-κB (NF-κB) were analyzed using quantitative Real-time PCR experiments. Moreover, histopathological and stereological changes of kidney and its related structures were investigated for the studied rats. Bip, Chop, and NF-κB expression levels were significantly decreased under bilirubin treatment, while sXbp1 was up-regulated following the bilirubin administration. More interestingly, glomerular constructive damages seen in HFD-T2D rats, were considerably improved in the animals received bilirubin. Stereological assessments also revealed that bilirubin could desirably reverse the mitigation of kidney's total volume and its related structures, such as cortex, glomeruli, and convoluted tubules. Taken together, bilirubin has potential protective/ameliorative effects on DKD progression, especially through alleviating the renal ER stress and inflammatory responses in T2D rats with injured kidneys. In this era, clinical benefits of mild hyperbilirubinemia can be considered in human DKD.

Resveratrol ameliorates bisphenol A-induced testicular toxicity in adult male rats: a stereological and functional study.

BACKGROUND: Bisphenol A (BPA) is one of the most widely used synthetic chemicals worldwide. BPA as an endocrine disruptor affects the reproductive systems through estrogenic and antiandrogenic proprieties. Resveratrol (RES) as a natural polyphenol and potent antioxidant exhibits protective effects against reproductive toxicity by inhibiting of oxidative stress. 48 male rats were divided into eight groups (n=6), including CONTROL, OLIVE OIL (0.5 ml/ day), Carboxy methylcellulose (CMC) (1 ml of 10 g/l), RES (100mg/kg/day), low dose of BPA (25 mg/kg/day), high dose of BPA (50 mg/kg/day), low dose of BPA + RES, and high dose of BPA + RES. All treatments were done orally per day for 56 days. At the end of the 8th week, blood samples were collected for hormone assays. Then, the sperm parameters were analyzed, and the left testis was removed for stereological study. RESULTS: We showed a significant decrease in sperm parameters in the low and high doses of BPA groups compared to control groups (P<0.05). The volume of testicular components as well as the diameter and length of seminiferous tubules significantly reduced (11-64 %), and the total number of the testicular cell types decreased (34-67 %) on average in the low and high doses of BPA groups. Moreover, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone hormones concentration showed a significant reduction in both doses of BPA groups (P<0.01). Nonetheless, treatment with RES could ameliorate all the above-mentioned changes in the low and high doses of BPA groups (P<0.05). CONCLUSIONS: RES could prevent BPA-induced testicular structural changes and sperm quality via improving gonadotropin hormones and testosterone levels.

RèSUMè: CONTEXTE: Le bisphénol A (BPA) est l'un des produits chimiques synthétiques les plus utilisés dans le monde. Le BPA en tant que perturbateur endocrinien affecte le système reproducteur par le biais de ses propriétés œstrogéniques et anti-androgènes. Le resvératrol (RES), en tant que polyphénol naturel et puissant antioxydant, présente des effets protecteurs contre la toxicité sur la reproduction en inhibant le stress oxydatif. Quarante-huit rats mâles ont été divisés en huit groupes (n = 6), comprenant les groupes TÉMOIN, HUILE D'OLIVE (0,5 ml/jour), méthylcellulose Carboxyle (MCC) (1 ml de 10 g/L), RES (100 mg/kg/ jour), faible dose de 25 de BPA (25 mg/kg/jour), dose élevée de BPA (50 mg/kg/jour), faible dose de BPA + RES et dose élevée de BPA + RES. Tous les traitements ont été effectués quotidiennement par voie orale pendant 56 jours. À la fin de la 8ème semaine, des échantillons de sang ont été prélevés pour dosages hormonaux. Ensuite, les paramètres du sperme ont été analysés et le testicule gauche a été retiré pour une étude stéréologique. RéSULTATS: Nous avons montré une diminution significative des paramètres spermatiques dans les groupes traités par doses faibles et doses élevées de BPA par rapport aux groupe témoin (P<0,05). Le volume des composants testiculaires ainsi que le diamètre et la longueur des tubules séminifères ont été considérablement réduits (11-64 %) ; le nombre total des types de cellules testiculaires a diminué (34-67 %) en moyenne dans les groupes traités par doses faibles et doses élevées de BPA. De plus, la concentration sérique d'hormone folliculostimulante (FSH), lutéinisante (LH) et de testostérone a montré une réduction significative dans les groupes traités quelle que soit la dose de BPA (P<0,01). Néanmoins, le traitement par RES pourrait améliorer tous les changements mentionnés ci-dessus dans les groupes traités par doses faibles et élevées de BPA (P<0,05). CONCLUSIONS: Le RES pourrait avoir un effet positif sur les changements structurels testiculaires induits par le BPA, ainsi que la qualité du sperme, en améliorant les taux sériques d'hormones gonadotrophines et de testostérone. MOTS-CLéS: Bisphénol A Resvératrol Toxicité testiculaire Paramètres du Sperme Stéréologie.

A Quantitative Study on the Protective Effects of Resveratrol against Bisphenol A-induced Hepatotoxicity in Rats: A Stereological Study.

Background: Bisphenol A (BPA) is a widely used chemical with toxic effects on the liver. Resveratrol (RES) is an herbal compound with protective properties. This study aimed to investigate the protective effects of RES on the liver in rats exposed to BPA. Methods: This study was conducted in 2018 in Shiraz, Iran. Thirty Sprague-Dawley male rats were divided into five groups: a control group (distilled water), a sham group (olive oil as a BPA solvent), a BPA group (50 mg/kg), an RES group (100 mg/kg), and a RES+BPA group (50 mg/kg+100 mg/kg). Olive oil, BPA, and RES were administered to the animals via gavage for eight weeks. After eight weeks, the animals' livers were removed, and stereological measurements were made to obtain the total liver volume, portal triad volumes, hepatocyte nucleus and cytoplasm volumes, hepatocyte numbers, sinusoidal space volumes and lengths, and Kupffer cell (KC) numbers. The data were analyzed using the one-way analysis of variance test. Results: The hepatocyte number, the total liver volume, and hepatocyte nucleus and cytoplasm volumes in the BPA group decreased by 41% (P<0.001), 18% (P<0.001), 32% (P=0.030), and 37% (P=0.014), respectively. The number of KCs and the length of sinusoids in the BPA group were increased compared with all the other groups (P<0.001). Our histological study revealed vacuolization, sinusoidal space dilatation, and congestion in the BPA group. Conclusion: In this study, the RES group, compared with the BPA group, exhibited a decrease in the total volume and length of sinusoids and the number of KCs. Additionally, the RES group showed an increase in the total liver volume, hepatocyte nucleus and cytoplasm volumes, portal triad volumes, and hepatocyte numbers after oral administration.

Protective Effect of Quercetin on Testis Structure and Apoptosis Against Lead Acetate Toxicity: an Stereological Study.

Exposure to environmental pollutants tightly impacts on the male fertility. In the present study, we examined the toxic effects of lead acetate (Pb) on testicular structure and the possible effect of quercetin on mitigating these effects. The apoptotic changes in the testes were also studied by the TUNEL assay and changes in apoptosis-related gene (Bax, Bcl-2, and caspase-3) expression. Twenty-one male mice were randomly divided into 3 groups of control, Pb, and lead acetate + quercetin. Testicular weight, both absolute and relative, was higher in Pb-exposed mice in comparison with the control and Pb-quercetin groups. The increase in size of testis was related to the lumen and connective tissue in this group. Lead acetate induced different patterns in testicular cell number; as spermatogonia, spermatocyte, and Sertoli cells number did not affect in lead acetate exposed group, while total number of round spermatids and long spermatids significantly reduced. In addition, Bcl-2 expression was downregulated, and Bax expression was upregulated in Pb-treated group in comparison with the control and Pb + quercetin groups. The TUNEL assay revealed that the number of apoptotic cells in Pb-treated group were increaed significantley in comparison to other groups. In conclusion, Pb administration adversely impacted on the cellular organization and activation of the apoptotic pathways in the testis; on the other hand, quercetin co-administration with lead partially ameliorated these adverse effects.

Effects of Different Storage Media, Temperature, and Time on Osteoblast Preservation in Autogenous Bone Grafts: A Histomorphometrical Analysis.

STATEMENT OF THE PROBLEM: Autogenous bone graft is the gold standard for bone reconstruction. Osteogenic cells must be kept viable in graft for a successful procedure. In extracorporeal preservation of grafts during surgery, three different factors may influence the quality of grafts. These factors include temperature, storage medium, and time interval. PURPOSE: In this study, we evaluated the effects of different storage media, temperatures, and times on osteoblast count in autogenous bone grafts, preserved extracorporeally. MATERIALS AND METHOD: Samples were obtained from iliac crest region in a goat. The grafts were preserved in 36 groups of different storage time, temperature, and medium. Samples were histomorphometrically analyzed to determine osteoblast count as the criteria of graft quality. RESULTS: In almost all samples, room temperature was the most and incubator was the least favorable storage temperatures. In grafts preserved in room temperature, no difference was noted between normal saline and Ringer's lactate solution and in almost all of the samples autologous blood and dry environment were more favorable media than Ringer's lactate solution. The effect of storage time was highly depended on the combination of temperature and solution. CONCLUSION: The results demonstrated that for preserving as many osteoblasts as possible in bone grafts, the best temperature was room temperature and the least favorable temperature was incubator. In addition, when bone fragments were preserved in room temperature, the best medium for graft storage was blood, which showed better results than normal saline and Ringer's lactate solution.

Ameliorative effects of quercetin on folliculogenesis in diabetic mice: a stereological study.

A high risk of reproductive disorders can be seen in diabetic pregnancy. Reproductive disorders associated with diabetes may result from alterations in the function of the ovary. In this study, we investigated the ameliorative effects of quercetin as a phytoestrogen and antidiabetic agent on the folliculogenesis in diabetic mice. Streptozotocin-induced diabetic mice were treated with 30 mg/kg/day quercetin for four weeks. The volume of ovary, follicles, and corpus luteum were significantly decreased in the diabetic mice. The number of growing follicles (secondary, antral, and Graafian follicles) and corpus luteum was significantly decreased in the diabetic mice. Also, the volume of oocytes was significantly decreased in antral and Graafian follicles. Our results indicated that the administration of quercetin in diabetic mice increased the volume of the ovary and growing follicles, the number of growing follicles and corpus luteum. It also significantly decreased the number of atretic follicles. As a result, it may be concluded that the impaired follicular growth and development caused by hyperglycemia in diabetic mice can be alleviated by quercetin treatment.

Apoptosis, Autophagy, and Necrosis in Murine Embryonic Gonadal Ridges and Neonatal Ovaries: An Animal Model.

BACKGROUND: In mammalian ovaries, loss of over two-thirds of germ cells happens due to cell death. Nonetheless, the exact mechanism of cell death has yet to be determined. The present basic practical study was designed to detect 3 types of programmed cell death, namely apoptosis, autophagy, and necrosis, in murine embryonic gonadal ridges and neonatal ovaries. METHODS: Twenty gonadal ridges and ovaries from female mouse embryos 13.5 days post coitum and newborn mice 1 day postnatal were collected. The TUNEL assay was performed to evaluate apoptosis. The interplay of autophagy was evaluated by immunohistochemistry for beclin-1. Necrotic cell death was analyzed by propidium iodide (PI) staining. The count and percentage of the labeled oocytes in the gonadal ridges and ovaries were evaluated and compared using the independent t test and one-way ANOVA. A P value less than 0.05 was considered statistically significant. RESULTS: We detected TUNEL-positive reaction in the embryonic germ cells and in the small and large oocytes of the neonatal ovaries. The germ cells and small oocytes reacted to beclin-1. PI absorption was detected in the embryonic germ cells and the large oocytes of the neonatal ovaries, but not in the small oocytes. The percentage of the TUNEL-positive and PI-labeled oocytes in the gonadal ridges was significantly higher than that in the neonatal ovaries (P<0.01 and P=0.01). In the neonatal ovaries, the percentage of the beclin-1-labeled oocytes was significantly higher than that in the embryonic phase (P<0.01). CONCLUSION: We showed that all 3 types of programmed cell death, namely apoptosis, autophagy, and necrosis, accounted for embryonic and neonatal germ-cell loss. Our observations demonstrated a potential role for necrosis, particularly in the embryonic gonadal ridge in comparison to the neonatal ovary, in mice.

The use of platelet-rich plasma (PRP) to improve structural impairment of rat testis induced by busulfan.

Platelet-rich plasma (PRP) has a potential effect on tissue repair through proliferation and differentiation of tissue progenitor cells. The aim of this study was to evaluate the effect of PRP on the testis structure and function in infertile rat model by stereological method. Thirty-two male rats were divided into four groups. Infertility was induced by the administration of busulfan (BUS) (10mg/kg, I.P., single dose). PRP (80µl, testis local injected, single dose) was administered for the subjects. After 48 days, semen analysis was performed and blood samples were taken from the heart to measure the testosterone. Then, the left testis was removed, fixed, embedded, sectioned, and stained by H&E; after that the testes were analyzed. The results showed that BUS can decrease the sperm count, motility, normal morphology, length of the spermatozoon tail, volume of the testis, seminiferous tubules, germinal epithelium height, and the number of spermatogenesis lineage cells in comparison with the control group (p < 0.01). PRP increased the number of spermatogenic stem cell, count, motility and tail length of the sperm and testosterone level in BUS-treated animal significantly, but it did not have any effect on the volume of the testis, germinal epithelium height, Sertoli and Leydig cells number, and seminiferous tubules length. It concluded that PRP can improve the structural and functional impairment of the testis in treatment by BUS.

Effect of platelet-rich plasma (PRP) on ovarian structures in cyclophosphamide-induced ovarian failure in female rats: a stereological study.

BACKGROUND: Ovarian failure is diagnosed by ovarian destruction and reducing sex hormonal levels. Platelet-rich plasma (PRP) contains several growth factors that induce tissue repair and may induce folliculogenesis. OBJECTIVE: This study evaluates the effect of PRP on ovarian structures and function in cyclophosphamide (Cy)-induced ovarian failure in female rats by a stereological method. METHODS: Thirty-two adult female rats were divided into four groups (Control, Cy, Cy + PRP, and PRP). Female infertility was induced by Cy (75 mg/kg, single dose, intraperitoneally). Animals were treated by PRP which was obtained from age-matched male rats (200 µL, single dose, intraperitoneally). Blood samples were collected for measurement of hormones. The animals were dissected and the right ovaries were removed, fixed, sectioned, and stained by H&E. Stereological methods were used to estimate cortex and medulla volume, and the number and diameter of follicles, follicular cell, and oocyte using light microscopy. RESULTS: Cyclophosphamide had the maximum effect in decreasing on cortex volume, the pre-antral follicles number, a diameter of follicular cells and oocyte diameter in the antral follicle and the reduction of estradiol and progesterone levels compared with the control group. PRP had a dominant positive effect on the ovarian cortex volume, pre-antral follicles number and antral follicle diameter relative to the control group. PRP also decreased oocyte diameter in pre-antral follicle in infertile animals (p < 0.001). CONCLUSION: It seems that PRP has a protective effect on ovarian failure in the infertile female rat model.

The non-preventive effects of human menopausal gonadotropins on ovarian tissues in Nandrolone decanoate-treated female rats: A histochemical and ultra-structural study.

BACKGROUND: The follicular growth and development may be affected by abused drugs. Nandrolone decanoate (ND) as an anabolic androgenic steroid can damage the morphological and functional features of the ovary and may lead to reproductive failure. OBJECTIVE: This study was designed to evaluate the effects of synchronized and non-synchronized administration of Human Menopausal Gonadotropins (hMG) with ND on ovarian tissue and level of sex hormones in the adult female rat. MATERIALS AND METHODS: Forty adult female Sprague Dawley rats were divided into eight groups. The five experimental groups received 3 and/or 10 mg/kg of ND synchronized and non-synchronized with 10 IU of hMG and hMG alone. The two shams and control groups received solvents of ND and hMG. The animals' serum levels of Follicle-stimulating hormone, Luteinizing hormone, progesterone and estrogen and the weight, volume and dimensions of the ovaries were measured. The ovaries were prepared for apoptosis assessment and morphological study. RESULTS: The ovarian volume and sex hormones in the experimental groups were decreased, but ovarian weight and dimensions didn't change. The rate of apoptosis was increased in the experimental groups as follows; a low and high dose of ND synchronized with hMG 48.80±18.70 and 65.20±14.20 respectively vs. Sham 1, 33.20±17.80, a low and high dose of ND non-synchronized with hMD 55.80±17.20 and 75.20±14.30 respectively vs. Sham 2, 31.60±32.40 groups, p˂0.01. Follicular and stromal cells were damaged in the experimental groups except for the hMG group. CONCLUSION: Administration of ND decreased the serum level of Luteinizing hormone, Follicle-stimulating hormone, progesterone and estrogen and damaged ovarian tissue irreversibly and irreparably and hMG cannot prevent the destruction of the follicles in the adult female rats. This can be a serious warning to women who abuse ND.

Modulatory effect of gonadotropins on rats' ovaries after nandrolone decanoate administration: a stereological study.

BACKGROUND: Nandrolone decanoate (ND) is an anabolic androgenic steroid (AAS) which influences the ovarian structure and function. We assessed the effects of ND on the ovarian volume, number of primordial follicles, and level of hormones and also evaluated the modulatory effects of gonadotropins on the histopathological changes imposed by the administration of ND. METHODS: Six groups of Sprague-Dawley adult female rats (n=30) were used. The experimental rats were injected intraperitoneally with 3 and 10 mg/kg ND with or without human menopausal gonadotropin (hMG), 10 IU weekly for one month. The vehicle and control rats were administered olive oil and saline, respectively, for the same period of time. The ovarian volume and number of primordial follicles were estimated by stereological methods. RESULTS: The results showed a decrease in the ovarian volume, number of primordial follicles, and level of gonadotropins in the ND-treated animals compared with the vehicle groups. In the rats treated with 3 mg/kg of ND with hMG, an increase in the ovarian volume and number of primordial follicles was shown as compared to the rats treated with the same dose of ND without hMG. CONCLUSION: ND exerted detrimental effects on the dimensions of the ovary, number of follicles, and level of sex hormones. However, hMG, prevented the harmful effects of ND (at least in a low dose) on the ovarian follicles.

Stereological study of the effect of ginger's alcoholic extract on the testis in busulfan-induced infertility in rats.

BACKGROUND: In traditional medicine zingiber officinale used to regulate female menstural cycle and treat male infertility. Recent studies have suggested the possible role of ginger extract in improving the testicular damage of busulfan. OBJECTIVE: The aim of this study was to evaluate the effects of zingiber officinale on the sperm parameters, testosterone level and the volume of the testes and seminiferous tubules by stereological methods. MATERIALS AND METHODS: Fifty rats were divided into four groups. All the rats were given a single intraperitoneally injection of 5mg/kg busulfan solution. The first group was kept as busulfan control, while the other groups were orally administrated ginger extract in graded doses of 50, 100 and 150mg/kg b.wt, for 48 consecutive days. At the end, all animals were anesthetized and their testes and vas deference were removed, fixed, embedded, and stained. The volume of testes and seminiferous tubules were estimated by cavalieri methods. RESULTS: The result showed, that zingiber officinale increased the volumes of seminiferous tubule in 100mg/kg treated group compared to control group. Sperm count (706×10(5) and 682×10(5)) and the level of testosterone (50.90 ng/mL and 54.10 ng/mL) enhanced in 100 mg/kg and 150 mg/kg treated groups compared to control group (p=0.00). CONCLUSION: It seems that zingiber officinale stimulate male reproductive system in induce busulfan infertility.