Testicular Biopsy Score Count Research Articles

Sequential Administration of Febuxostat, Amlodipine and Vitamin E Attenuate Oxidative Stress and Improve Spermatogenesis in Testicular Ischemia Reperfusion Injury in Wistar Rats

This study investigated the effects of sequential administration of febuxostat, amlodipine, and vitamin E on oxidative stress and spermatogenesis following testicular ischemia-reperfusion injury (TIRI) in Wistar rats. Ninety male rats (120-150 g) were divided into 9 groups (n=10 each): sham operation (SO), torsion-detorsion (TD), and seven treatment groups receiving different combinations of the drugs. The treatment groups included torsion + febuxostat + detorsion (TFD), torsion + detorsion + amlodipine (TDA), torsion + detorsion + vitamin E (TDV), and combinations thereof (TFDA, TFDV, TDAV, TFDAV). TIRI was induced by 720° clockwise testicular torsion for 1 hour followed by detorsion. Febuxostat (5 mg/kg) was administered 30 minutes after torsion, amlodipine (2.5 mg/kg) immediately upon detorsion, and vitamin E (10 mg/kg) 30 minutes after detorsion. Rats were sacrificed 56 days post-reperfusion. Testicular tissue was analyzed for antioxidant enzymes (superoxide dismutase, catalase), lipid peroxidation (malondialdehyde), total protein, inflammatory markers (serum nitrite, IL-1β), and spermatogenesis indices (testicular biopsy score, Leydig cell count). Results showed that TIRI significantly decreased antioxidant enzymes, significantly increased lipid peroxidation and inflammatory markers, and impaired spermatogenesis. In the TD group, superoxide dismutase (SOD) and catalase (CAT) activities were significantly decreased, while malondialdehyde (MDA) increased significantly compared to the SO group (p<0.01). Serum nitrite and IL-1β levels in the TD group were also increased (p<0.001). Furthermore, the testicular biopsy score and Leydig cell count in the TD group were significantly decreased in this study (p<0.01). Sequential administration of febuxostat, amlodipine and vitamin E, particularly when all three were used (TFDAV group), significantly attenuated these changes. In the TFDAV group, SOD and CAT activities were improved, while MDA decreased compared to the TD group (p<0.05). Serum nitrite and IL-1β levels in the TFDAV group were also decreased (p<0.001). In addition, the testicular biopsy score and Leydig cell count in the TFDAV group increased in comparison with the TD group (p<0.001). The study concludes that this sequential multi-drug approach shows promise in mitigating the long-term detrimental effects of TIRI on testicular function and fertility. The combination of febuxostat (a xanthine oxidase inhibitor), amlodipine (a calcium channel blocker), and vitamin E (an antioxidant) appears to provide protection against oxidative stress and inflammation induced by TIRI, thereby preserving spermatogenesis.

Testicular Biopsy Score Count After Vasectomy in Albino Rats

Context: One of the challenging problems faced by the entire world is population explosion. The vasectomy is simple, safe, quick and effective vital method of male sterilization. In the recent years a large number of vasectomies have been performed in India, since the inception of National Family Planning Program in 1956. Thus the number of request for the restoration of fertility is also increased. Aim: The present study was carried out to know whether the testicular biopsy score count, indicating the process of spermatogenesis remains normal after vasectomy. Material & Methods: The testicular biopsy count of testes of 50 male albino rats was studied one, two, three and four months after vasectomy, in 10 male albino rats of each case and 10 male albino rats served as control. Result: After vasectomy seminiferous tubules are cut in various planes covered externally by connective tissue stroma with normal microscopic picture. The spermatogonia, spermatocytes, spermatids, Sertoli cells and spermatozoa show normal structure with normal size blood vessels in interstitial connective tissue, in vasectomised and control rats. In some seminiferous tubules granular cytoplasmic vesicle like structures are seen filling the lumen and spermatids in metamorphosis phase to spermatozoa. Many round or elongated heads of spermatozoa are present at apical portion of Sertoli cells. The control rats as well as vasectomised rats at four after vasectomy have shown the testicular biopsy score count of nine (9) and ten (10) on TBSC of Johenson. Conclusion: On critical analysis it is concluded that the testicular biopsy score count remains normal at four months after vasectomy.

Germinal epithelium changes in sexually immature rabbits treated with intratesticular testosterone implants: Samenepithel-Veränderungen bei unreifen Kaninchen nach Behandlung mit intrascrotalen Testosteron-Implantaten

The effect of testosterone (T) on germinal epithelium development and maturation was studied in prepubertal male rabbits. For this purpose, implants of free T (50 mg) or placebo (PL) were placed intratesticularly in 55-60-day-old animals. Each group was divided into sub-groups additionally treated with 0.9% NaCl, FSH/LH (3.0 IU) or cyproterone acetate (CA, 2.5 mg) over a 45-day period. Initial and final evaluations included measurements of testicular volume, testicular biopsy score count (TBSC) and plasma T, FSH and LH concentrations. At the end of the experiment, increment values (final-initial evaluation) of the parameters examined showed the following differences: (1) mean TBSC in T-implanted, irrespective of additional treatment, was higher than that of PL-implanted rabbits (P less than 0.01); (2) mean plasma T, FSH and LH values were significantly different in T- and PL-implanted rabbits, with higher T (P less than 0.05) but lower FSH and LH increments (P less than 0.01 for both) in the former group; (3) marked differences amongst types of additional treatment, irrespective of the implant used, were found for TBSC, T, FSH and LH (P less than 0.01 for all); animals treated with CA had markedly lower increments than other treatment groups. From these findings, it may be concluded, that in the prepubertal rabbit T plays an important role in development and maturation of the testis, but this effect probably requires the synergistic action of endogenous gonadotrophins.

Torsion-induced injury in rat testes does not affect mitochondrial respiration or the accumulation of mitochondrial mutations.

Male rats were subjected to 1 h testicular torsion of the spermatic cord or 1 h torsion followed by detorsion and recovery up to 4 weeks. The extent of tissue damage was evaluated by a testicular biopsy score count and mitochondrial function. Torsion for 1 h followed by detorsion induced significant morphological damage, which became more severe with longer periods of recovery. This morphological damage could not be correlated with mitochondrial damage as assessed by measuring the 4834 bp mitochondrial DNA 'common deletion' using a quantitative competitive polymerase chain reaction (PCR) assay. Mitochondrial respiratory chain activity, as measured by mitochondrial oxygen consumption using an oxygen electrode, did not vary between the treated animals and the controls. We conclude that the common mitochondrial DNA deletion and oxygen consumption are not good indicators of testicular damage induced by torsion.

Relationship between testicular morphology and sperm production following ischaemia in the ram

Arteriosclerosis was induced in the internal spermatic artery of rams to determine if this condition is implicated in the aetiology of testicular pathology which causes male infertility. Data were collected on sperm concentration and motility for 56 days following surgery to provide an index of testicular function. Testes were then weighed and a testicular biopsy score count was performed on histological sections to assess spermatogenic potential of seminiferous tubules. Vascular disturbance caused focal damage of the seminiferous epithelium, similar to that seen among infertile men, and a reduction in ejaculate volume, sperm concentration and sperm motility. Sperm concentration decreased following ischaemia yet was maintained to some degree by a germ-cell depleted spermatogenic epithelium. Normal testicular morphology was maintained above a testis weight of about 120 g (for an individual testis), but below this threshold spermatogenesis was severely impaired. In conclusion, these data have provided information on the relationship between testicular morphology and function following ischaemia in the ram. Furthermore, the morphological changes induced in the testis were similar to those seen among infertile men and, by their focal nature, could explain the distinction between oligozoospermia and azoospermia in men exhibiting spermatogenic arrest.

Effects of induced peripheral anosmia on gonadal maturation in prepubertal male rabbits.

The effect of induced peripheral anosmia on gonadal development and maturation was investigated in sexually immature male rabbits. Furthermore, the effect of agents promoting gonadal maturation (LHRH, hMG or methyl testosterone) on testicular development was examined in anosmic rabbits. Peripheral anosmia was induced by spraying the olfactory mucosa with 5% ZnSO4 solution; its effects were evaluated after a 45-day period, corresponding to the duration of spermatogenesis. Evaluation was based on measurement of body-weight, testicular size, testicular biopsy score count (TBSC) and a standard LHRH test (0 and 30 min) involving measurement of the blood levels of FSH, LH and testosterone before and at the end of the test. Markedly lower final and incremental values were noted in anosmic, compared to intact, animals for body-weight (P less than 0.001), TBSC (P less than 0.001), FSH (P less than 0.01) and LH (P less than 0.05). On the other hand, treatment of the anosmic rabbits with 0.9% saline, resulted in lower FSH, TBSC and testicular size increments than in rabbits treated with LHRH, hMG or testosterone, while testosterone levels and body-weight increments were similar in all groups. These findings indicate that induced peripheral anosmia is probably responsible for the inadequate gonadal maturation in prepubertal anosmic male rabbits. This relationship was confirmed by the observed stimulatory effect of administration of agents activating pituitary gonadotrophin secretion or gonadal function in anosmic animals.

Histologic Analysis of Orchiopexy in a Cryptorchid Rabbit Model

Although there is a consensus that very early orchiopexy is advantageous, there is little experimental data to support this view. Therefore, we studied early pre-pubertal versus later pre-pubertal orchiopexy in a cryptorchid rabbit model. Male offspring were divided into four groups with six to nine rabbits in each group: 1) controls (CON); 2) shams undergoing sham operations at both three weeks and two months of life (SHAM); 3) cryptorchid on day 3 with orchiopexy at three weeks and sham operation at two months (03W); 4) cryptorchid on day 3 with sham operation at three weeks and orchiopexy at two months (02M). All animals were sacrificed post-puberty at six months of age; the testes were fixed in situ by perfusion fixation, embedded in glycol methacrylate, and underwent morphometric analysis in a blinded fashion. The average seminiferous tubular diameter (STD) of the CON, SHAM, 03W, and 02M groups was 234, 160, 116, and 123 μm., respectively. The average testicular biopsy score count (TBSC) of the CON, SHAM, 03W and 02M groups was 9.0, 5.2, 3.5 and 3.5, respectively. There is no significant difference in STD or TBSC between the 03W and 02M cryptorchid groups, and values for both of these groups were significantly different from those of the SHAM group. In addition, a significant effect of sham operation was demonstrated. These results demonstrate no advantage of very early orchiopexy in this model, and are consistent with the clinical literature which, in general, shows reduced reproductive potential of a cryptorchid testis after orchiopexy. (J. Urol, 142: 413-417, 1989)

Testicular biopsy of azoospermic men with vas deferens malformation using two different techniques.

A morphological analysis was carried out on testicular biopsies of men with obstructive azoospermia as consequence of vas deferens malformations as compared with biopsies from fertile men. For each case the biopsies were processed with two different techniques: routine histological procedure, and semithin sections of specimens processed for electron microscopy. Four parameters were considered: tubular morphology, testicular biopsy score count (TBSC), tubular diameter, and germinal cell density. The data were quantified and analyzed by statistical tests. The biopsies of azoospermic men present a higher frequency of tubular sections with "tubular blockage," "sloughing" and cellular degeneration, lower values of TBSC, and lower germinal cell density. The results appear to be of relevant interest in relation to the existence of long-term testicular alteration following vasectomy.

Effect of experimental microvascular orchidopexy on spermatogenic cell differentiation in the testes of the dog.

Experimental microsurgical orchidopexy was undertaken in 15 healthy mongrel dogs by anastomosing the testicular artery and vein to the ipsilateral caudal epigastric vessels. At pre-selected post-operative intervals of 4 weeks, 5 or 8 months the results were morphologically and histologically evaluated by the 'testicular biopsy score count' method. This method is based on the presence of the main spermatogenic types arranged in order of maturity. Between the mean scores of the normal and the operated testes there was a significant difference. The differentiation of the spermatogenic epithelium of the reconnected testis is slightly hampered, but not seriously.

Value of a testicular biopsy rating for prognosis in oligozoospermia.

Testicular biopsies of 142 oligozoospermic men were used to obtain a testicular biopsy score count. These scores were clearly related to the chance of fertility, in contra-distinction to data on hormone concentrations and from analysis of semen. In 36 patients with a score of 9-10 there were 15 pregnancies; in 59 patients with a score of 8-9, 12; and in 47 patients with scores below 8 there was only one successful pregnancy, though this last group also contained an additional three patients whose wives' pregnancies ended in miscarriages. Rating of testicular biopsies is more useful for proper evaluation of oligozoospermic patients than are data on hormone concentrations and from semen analysis. Proper evaluation of new treatments should be based on trials where experimental and control groups are matched on these ratings.

A relationship between testicular biopsy score count and fertility of men treated for oligospermia

A relationship between testicular biopsy score count and fertility of men treated for oligospermia

Studies on the relationship between spermatogenesis and urinary levels of follicle stimulating hormone and luteinizing hormone in oligospermic men.

ABSTRACT The urinary excretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH) was measured by specific bioassays in 122 infertile men from barren couples in order to study the relationship between levels of FSH and LH and the state of the germinal epithelium. The latter was assessed both by determination of the gametic output and by examination of testicular histology, using the testicular biopsy score count method, in which the mean score (MS) is the overall measure of level of spermatogenesis (Johnsen 1970a). A highly significant negative correlation was established between log FSH and both log sperm concentration and MS and also between log LH and MS, but not between log LH and log sperm concentration. Of the 122 infertile men, the 56 who belonged to the heterogeneous idiopathic oligospermia group were studied most intensely. Nineteen of these had elevated urinary FSH levels. The mean FSH level for this group was 2.3 times higher than the mean FSH level for normal males (P < 0.001). Only 5 patients in this group had an elevated urinary LH level and the mean LH level was not different from that of normal men. In this idiopathic oligospermia group there was a significant negative correlation between log FSH and MS, and between log FSH and sperm concentration, but no correlation between log LH and either of the same 2 parameters. No correlation between log FSH and log LH on the one hand and the excretion of oestrogens, androgen metabolites, sperm morphology or motility on the other hand was found. The excretion of testicular and adrenal androgen metabolites (separated by a dexamethasone-suppression test) was normal in the 56 males with idiopathic oligospermia. It is concluded that there is a close relationship between urinary FSH levels, and to a lesser degree urinary LH levels, and spermatogenesis and that testicular histology is a better parameter for the judgment of the state of the germinal epithelium than the sperm concentration. It is further concluded that analyses of gonadotrophins, especially of FSH, are of value in the differential diagnosis of the infertile man, especially in distinguishing between those who will benefit from a treatment with gonadotrophins and those who will not.

A Relationship Between Testicular Biopsy Score Count and Fertility of Men Treated for Oligospermia

A Relationship Between Testicular Biopsy Score Count and Fertility of Men Treated for Oligospermia

Evaluation of gonadotrophin analyses in male hypogonadism.

ABSTRACT Urinary total hypophyseal gonadotrophin (HG) analyses in 56 healthy men and 282 patients with a great variety of forms of male hypogonadism showed that in some clinical conditions HG is decreased, in others increased while in some others the HG level is normal. It has been shown, however, that primary damage of the germinal epithelium per se causes an increase of the HG level. It is necessary, therefore, to take the functional state of the germinal epithelium into account when HG excretion is used as a parameter of function and responsiveness of the hypophysis. By the testicular biopsy score count method described by the author, a figure (mean score, MS) is obtained which expresses the spermatogenetic state. Analysis of 284 patients with primary testicular disorders established a correlation between the log gonadotrophin value (log HG) and MS. It was found that the HG level in the control group of apparently healthy men was considerably higher than that corresponding to perfect spermatogenesis. The question whether »ideal« men with perfect spermatogenesis or »average healthy« men should be used as controls in gonadotrophin assays is a dilemma. By using the regression equation between log HG and MS the influence of various spermatogenetic states on the HG level was eliminated by transforming, in each patient, the log HG value to the value corresponding to a fixed mean score of 1.0. These transformed log HG values were then evaluated in the various disorders. Decreased values were found in hypopituitarism, infantilism, eunuchoidism, adiposogenital dystrophy (in adults) and hypogonadism secondary to metabolic disease. In all other conditions, the HG level corresponded to the spermatogenetic state. No extra-tubular factors seem to influence the HG level and no case of primary overproduction of gonadotrophins was found. The value of transforming gonadotrophin values to a defined spermatogenetic state in clinical and experimental studies is discussed.

The stage of spermatogenesis involved in the testicular-hypophyseal feed-back mechanism in man.

ABSTRACT In 284 patients with a great variety of primary testicular disorders registration of spermatogenesis was performed with the testicular biopsy score count method described by the author. Correlation analyses between the score count steps and gonadotrophin excretion show a great rise when spermatozoa (late spermatids) disappear from the testis. There is a high correlation between the per cent of tubuli possessing spermatozoa and the gonadotrophin level. Disappearance also of spermatids or spermatocytes or spermatogonia induces no further rise in gonadotrophins and no separate effect of these cells on gonadotrophins could be demonstrated. Some influence upon the gonadotrophin level was found when only Sertoli cells were present but this was small as compared with the effect of spermatozoa. It is concluded that the stage of spermatogenesis involved in the testicularhypophyseal feed-back mechanism in man is the last stage where the spermatozoon is finished and matured. The way this stage exerts its influence upon the gonadotrophin level is briefly discussed.

Testicular biopsy score count--a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males.

The paper describes a new and rapid method forregistration of spermatogenesis in human testes: the testicular biopsy score count. Each tubular section is given a score from 10 to 1 according to presence or absence of the main cell types arranged in the order of maturity. Presence of spermatozoa scores 10, 9 or 8; spermatids (and no further) 7 or 6; spermatocytes (and no further) 5 or 4; only spermatogonia 3, only Sertoli cells 2 and no cells 1. The theoretical background of the score count method is discussed and it is emphasized that tissue heterogeneity, being a main point in most conditions, is exposed and evaluated by the method. Normal values are given and results obtained in 335 cases including a great variety of forms of male hypogonadism are presented. Patognomonic score counts leading to immediate diagnosis at a glance are obtained in many instances. A high correlation between testicular biopsy score count and sperm count is found and it is concluded that by this method it has for the first time become possible in man to correlate endocrine conditions with the functional state of the testicular tissue.