Effect of Zirconium oxide nanoparticle on serum level of testosterone and spermatogenesis in the rat: An experimental study.

BACKGROUND: Zirconium nanoparticles are used as health agents, pharmaceutical carriers, and in dental and orthopedic implants. OBJECTIVE: This studyaimed to investigate the effects of Zirconium oxide nanoparticles on the process of spermatogenesis in rat. MATERIALS AND METHODS: In this experimental study, 32 male Wistar rats (150-200 gr), with range of age 2.5 to 3 months were used and divided into four groups of eight per each. The control group received 0.5 ml of distilled water and the three experimental groups received 50, 200, and 400 ppm doses of Zirconium oxide nanoparticles solution over a 30-day period, respectively. At the end of the experiment, tissue sections were taken from the testis and stained with hematoxylin-eosin. Serum concentration of testosterone was measured by enzyme-linked immunosorbent assay. RESULTS: In the experimental group receiving 400 ppm Zirconium oxide nanoparticles, the number of Spermatogonia cells (p ≤ 0.01), Spermatocytes (p ≤ 0.01), Spermatids (p ≤ 0.001), and sertoli and Leydig cells (p ≤ 0.05) showed a significant decrease compared to the control group. Serum testosterone concentration did not change significantly in all experimental groups receiving Zirconium oxide nanoparticles compared to the control group. Experimental group received 400 ppm Zirconium oxide nanoparticles shrinkage of seminal tubules and reduced lumen space compared to control group. CONCLUSION: Zirconium oxide nanoparticles are likely to damage the testes by increasing Reactive oxygen species production and free radicals.

Adipose tissue-derived mesenchymal stem cells repair germinal cells of seminiferous tubules of busulfan-induced azoospermic rats.

CONTEXT: Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are less invasive than bone marrow mesenchymal stem cells to obtain for cell therapy. AIMS: The aims of this study were to evaluate the germinal cells characteristics and repairs in seminiferous tubules of busulfan-induced azoospermic rats after AT-MSCs transplantation. SETTINGS AND DESIGN: Experimental case-control study. MATERIALS AND METHODS: In the present experimental study, donors AT-MSCs were isolated from subcutaneous adipose tissue of two Sprague-Dawley rats. The recipients (n = 5) were received two doses of 10 mg/kg of busulfan with 21 days interval to stop endogenous spermatogenesis. After induction of azoospermia by busulfan, rats were injected with the AT-MSCs into the efferent duct of right testes. After 60 days, the right testes were injected AT-MSCs were compared to left azoospermic testes. Five untreated male rats served as negative control. STATISTICAL ANALYSIS USED: Stereological indices were analyzed by one-way ANOVA and LSD post-hoc test. The spermatogenesis index was compared using Mann-Whitney U test. RESULTS: After stereological analyses, the seminiferous tubules treated with AT-MSCs had normal morphology. The untreated seminiferous tubules were empty. Spermatogenesis was observed in most cell-treated seminiferous tubules. CONCLUSIONS: The testis of busulfan-induced azoospermic rats accepted transplanted AT-MSCs. The transplanted AT-MSCs could induce spermatogenesis in seminiferous tubules of the rat.

Busulfan induced azoospermia: Stereological evaluation of testes in rat.

The aim of the present study was stereological evaluation of testes of azoospermic animal model using busulfan in rat. Three groups of male adult rats were used in this study. The first group was injected by single dose of busulfan (10 mg kg(-1)) and their testes were removed on day 35 post injection. The second group received double doses of busulfan with 21 days interval and their testes were removed on day 35 after the second injection. The testes of the third group were removed without busulfan therapy. In 10 circular transverse sections of tubules stained with hematoxylin-eosin, stereological parameters were measured. The testes were rated for its spermatogenic potential on a modified spermatogenic scale of 0 to 6. Cellular (germinal epithelium) diameter and area of the seminiferous tubules, total diameter and cross sectional area of the tubules of the seminiferous tubules in rats that received double doses of busulfan were less than the rats in single dose of busulfan and control groups (p < 0.05). Spermatogenesis index of seminiferous tubules in rats receiving two doses of busulfan was less than the rats received one dose of busulfan (p < 0.001) and the index of both treatment groups were less than the control group (p < 0.001). In conclusion, two doses of busulfan injection with 21 days interval produced an appropriate experimental model of induced azoospermia with comparable stereological indices of seminiferous tubules in rat.

Computer-assisted design and synthesis of a highly selective smart adsorbent for extraction of clonazepam from human serum.

A computational approach was applied to screen functional monomers and polymerization solvents for rational design of molecular imprinted polymers (MIPs) as smart adsorbents for solid-phase extraction of clonazepam (CLO) form human serum. The comparison of the computed binding energies of the complexes formed between the template and functional monomers was conducted. The primary computational results were corrected by taking into calculation both the basis set superposition error (BSSE) and the effect of the polymerization solvent using the counterpoise (CP) correction and the polarizable continuum model, respectively. Based on the theoretical calculations, trifluoromethyl acrylic acid (TFMAA) and acrylonitrile (ACN) were found as the best and the worst functional monomers, correspondingly. To test the accuracy of the computational results, three MIPs were synthesized by different functional monomers and their Langmuir-Freundlich (LF) isotherms were studied. The experimental results obtained confirmed the computational results and indicated that the MIP synthesized using TFMAA had the highest affinity for CLO in human serum despite the presence of a vast spectrum of ions.

Computational design and synthesis of molecular imprinted polymers for selective extraction of allopurinol from human plasma.

The present study was focused on the rational development of polymers for selective extraction of allopurinol (ALP) from human plasma. Therefore, a computational modeling approach was combined with the molecular imprinting technology to obtain the polymers. The computational approach was used in order to screen the functional monomers as well as the polymerization solvents for rational design of molecular imprinted polymers (MIPs). It was based on the comparison of the binding energy (ΔE) of the formed complexes between the template molecule and different functional monomers. In the design, the effect of the polymerization solvent was also included using the polarizable continuum model. The theoretical calculation results showed that among virtual solvents tested, acrylamide (AAM) gave the largest ΔE while acrylonitrile (ACN) gave the smallest ΔE in acetone. Therefore, the MIP prepared using AAM as functional monomer in acetone was desired. To examine the validity of this approach, three MIPs were synthesized with different functional monomers i.e. AAM, acrylic acid (AA), and ACN, and then evaluated using Langmuir-Freundlich (LF) isotherm. The results obtained from this experiment confirmed the computational results that the MIP prepared by AAM was the most appropriate adsorbent. Subsequently, the MIP was used to develop a molecular imprinted solid-phase extraction (MISPE) procedure. Finally, the MISPE procedure followed by HPLC was developed for selective extraction and determination of allopurinol in human plasma. For the proposed MISPE method, the linearity between peak area and concentration was found in the range of 0.100-25.000 µM with a linear regression coefficient (R²) of 0.995. The limit of detection (LOD) and quantification (LOQ) in plasma were 0.028 and 0.093 µM, respectively. The results of this study indicated the possibility of using computer aided design for rational selection of functional monomers and solvents for preparation of the MIPs capable of extracting allopurinol from human plasma.