Effects of Benzo(a)pyrene on the endometrial receptivity and embryo implantation in mice: An experimental study

Background: Benzo(a)pyrene (BaP) as an environmental pollutant is ubiquitous in the environment and it has destructive effects on human health. So far, various studies have demonstrated that BaP can cause adverse effects on the female reproductive system, but the existing information is limited about the effects of BaP on the endometrial receptivity and embryo implantation. Objective: The aim of this study was to investigate the effects of BaP on the endometrial receptivity and implantation in mice. Materials and Methods: In this experimental study, 40 pregnant BALB/c mice were divided into 5 groups (n = 8/each) as follows: experimental groups received the doses of 100 μg/kg, 200 μg/kg, and 500 μg/kg BaP dissolved in corn oil, the control group received normal saline and sham group received corn oil. Pregnant mice administered these solutions from Day 1 to Day 5 of gestation by gavage. On Day 6, the mice were sacrificed. Then their embryos were counted and the hormonal, histomorphological and molecular analyses were performed on themocusa of uterine tube. Results: The data revealed that BaP reduces estrogen and progesterone levels, decreases the number of implantation site, endometrium thickness, uterine lumen diameter, stromal cells and endometrial glands, and blood vessels in the endometrium. However, the expression of Activin receptor-like kinase 5 and E cadherin genes was not changed by BaP with different doses. Conclusion: The finding of this study showed that BaP can change estrogen and progesterone levels, and endometrial morphology leads to impairing the endometrial receptivity and decreasing the number of implantation site. Key words: Benzo(a)pyrene, Embryo implantation, Estrogen, Progesterone, ALK5, E-cadherin

Protective effect of green tea extract on the deltamethrin-induced toxicity in mice testis: An experimental study

Background: Deltamethrin (DM) is one of the environmental factors that can have destructive effects on the male fertility. Green tea (GT) as a medicinal herb, has antioxidant property. Objective: The present study investigated the protective role of GT extract in improving the harmful effects of DM on the testis. Materials and Methods: In this experimental study, 35 adult male mice (25–30 gr) were divided in to five groups (n = 7/each). The control group received only normal saline. Sham received 0.2 ml corn oil. Green tea group received only GT of 150 mg/kg. bw; deltamethrin group received the DM at a dose of 0.6 mg/kg. bw; GT + DM received both GT and DM. The effect of GT was assessed by measuring oxidative stress markers, sperm parameters, histological and immunohistochemical analysis. Results: The results showed that the count and motility of spermatozoa, testosterone, and Malondialdehyde significantly decreased (p < 0.001) and the abnormal spermatozoa increased (p < 0.001) in DM group compared to control group. Moreover, enhanced caspase-3expression and apoptosis were observed in DM-treated mice compared to control group. Histologically, DM with a degenerative effect on testicular tissue reduced the spermatogenesis progenitor cells. The epithelial height and the diameter of the seminiferous tubules were also reduced in the DM group. Treatment with GT in the DM-treated mice significantly improved these changes. Conclusion: With these findings, it was concluded that the GT treatment with antioxidant activity and anti-apoptotic property could protect the testicular injury induced by DM

Homing of adipose stem cells on the human amniotic membrane as a scaffold: A histological study

Background: The human amniotic membrane (HAM) is a suitable and effective scaffold for cell culture and delivery, and adipose-derived stem cells (ADSCs) are an important source of stem cells for transplantation and chondrogenic differentiation. Objective: To assess the practicability of a cryopreserved HAM as a scaffold in cell proliferation and differentiation in vitro. Materials and Methods: In this experimental study, adipose tissue samples were harvested from the inguinal region of male patients aged 15-30 years. Flow cytometry was used to identify CD31, CD45, CD90, and CD105 markers in adipose stem cells. HAM was harvested from donor placenta after cesarean section, washed, trypsin-based decellularized trypsinized decellularized, and used as a scaffold via three methods: 1) ADSCs were differentiated into chondrocytes on cell culture flasks (monolayer method), and after 14 days of culture, the cells were transferred and cultured on both sides of the HAM; 2) ADSCs were cultured and differentiated directly on both sides of the HAM for 14 days (scaffold-mediated differentiation); and 3) chondrocytes were differentiated with micromass culture for 14 days, transferred on HAM, and tissue slides were histologically analyzed qualitatively. Results: Flow cytometry confirmed the presence of mesenchymal stem cells. Histological findings revealed that the cells adhered and grew well on the stromal layer of HAM. Among the three methods, scaffold-mediated differentiation of ADSCs showed the best results. Conclusion: ADSCs have excellent attachment, viability, and differentiation capacity in the stromal side of HAM. Additionally, the direct culture and differentiation of ADSCs on HAM is more suitable than the culture of differentiated cells on HAM. Key words: Amniotic membrane, Scaffold, Chondrogenesis, Differentiation, Mesenchymal stem cell

Growth and chondrogenic differentiation of mesenchymal stem cells derived from human adipose tissue on chitosan scaffolds

BACKGROUND AND OBJECTIVE: Treatment of cartilage damage for any reason is associated with temporary relief of joint pain. Providing the possibility of differentiating various stem cells into adult tissues can contribute to recovery and treatment of damaged cartilage tissue in skeletal system. In this study, chondrogenic potential of chitosan scaffold, CH-β-GP-HEC, with stem cells derived from human adipose tissue. METHODS: In this cross-sectional study, adipose tissue-derived stem cells were separated from abdomen of 15 patients who underwent inguinal hernia repair. 6-7×105 cells were cultured in plate one-dimensionally and on chitosan scaffold three-dimensionally for 21 days. MTT assay was run to evaluate the toxic effect of scaffold on cell viability. Proliferation and differentiation of cells were studied in the two types of culture after toluidine blue staining. To confirm the formation of cartilage, expression of collagen type II was assessed by immunohistochemistry. FINDING: In MTT assay, the average OD for cells cultured on scaffold is higher than 0. 8 compared with control group, which confirms the nontoxicity of scaffold for culturing stem cells (p>0. 05). Chondrogenic differentiation of cells on scaffold shows more glycosaminoglycan deposition in the extracellular matrix compared with one-layer culture. Moreover, in group with three-dimensional culture system, cells were spherical and the morphology of nucleus was different from one-layer culture. Regarding immunohistochemistry results, increased synthesis was observed in collagen type II as chondrogenesis markers in three-dimensional culture system compared with one-layer culture. CONCLUSION: Results of the study revealed that hydrogel scaffold, CH-β-GP-HEC, with porous structure provides a better environment for the growth of mesenchymal stem cells and their differentiation into cartilage tissue. © 2016, Babol University of Medical Sciences. All rights reserved

Comparison of conditioned medium and direct co-culture of human granulosa cells on mouse embryo development

189-192Although numerous investigations have demonstrated the beneficial effects of co-culture system of different somatic cells on in vitro development of embryos, the effects of conditioned-media of co-culture cells have not been well documented. The objective of this study was to compare the effects of human granulosa cells co-culture system and its conditioned medium on the developmental rate of mouse embryos in vitro. Two sets of experiments were undertaken; in the first one 317 mouse one-cell embryos were cultured in human granulosa cell co-culture system (GC), Ham’s F10 medium conditioned with granulosa cells (CM) and non-conditioned Ham’s F10 for 120 h. In the second experiment, 391 late two-cell embryos were cultured in the 3 fore-mentioned culture treatments for 72 h. Embryos were obtained from NMRI mice. Granulosa cells were collected from patients undergoing an IVF program during oocyte pickup. In the first set of experiments, 23.6, 14.5 and 11.1% of one-cell embryos passed two-cell block and continued growing to 4-cell in GC, CM and HF, respectively. This index in GC was significantly different from two other treatments. Also significantly more embryos reached blastocyst stage in GC compared with two other treatments. The blastocyst rate was not significantly different between CM and HF. In the second set of experiments the proportion of blastocyst stage was significantly higher in CM than that in HF and lower than that in GC. In conclusion, although human granulosa cell-conditioned medium has beneficial effects on mouse embryo development, it was not as effective as co-culture of these cells

Morphometrical study of mouse pre-implantation embryos: comparison of in vitro- and in vivo-produced embryos

(Received 15 Nov, 2008; Accepted 30 Jan, 2009)Abstract Background and purpose: The development of pre-implantation mammalian embryos in vitro is compromised, compared with those grown in vivo. Selecting embryos with a high implantation potential is one of the most important challenges in the field of assisted reproductive technology. The aim of this study was to postulate morphometrical characteristics of good quality embryos, with comparisons between in vivo and in vitro produced mouse embryos.Materials and methods: Embryos was obtained from NMRI female mice after super ovulation. In vivo developed 2-, 4- and 8-cell embryos; morulla and full blastocyst were isolated from mice on 18, 36, 52, 60, 72 and 96 hours after hCG administration respectively. Ham, s F10 medium was used for in vitro culture of embryos. External and internal diameter of embryos, zona thickness and number of cells in full blastocysts were evaluated and compared between in vivo and in vitro groups.Results: External and internal diameter and zone thickness in oocyte and zygotes were 99.9µm, 75.4µm and 4.9µm respectively. These values did not change prior to the blastocyst stage in both in vivo and in vitro groups; but in full blastocyst stage, the diameter of embryos significantly increased and zone thickness decreased compared to prior stages in both groups (P<0.01). The diameter of full blastocysts of in vivo group (116.5 µm) were significantly larger than those of in vitro group (104.3 µm, P<0.05). Moreover, the full blastocysts of in vivo group had significantly more blastomeres (49), compared to in vitro group (43, P<0.05). Additionally, cultured embryos reached full blastocyst at 110 hours after hCG administration, while in vivo condition the time frame was 96 hours.Conclusion: Based on the above results, embryo size and zona thickness can not predict embryo quality prior to blastocyst stage, however, in this stage; larger embryos and those that have more blastomere may show greater viability. J Mazand Univ Med Sci 2008; 18(67): 34-49 (Persian

Virgin olive oil ameliorates deltamethrin-induced nephrotoxicity in mice: A biochemical and immunohistochemical assessment

Objective: A major class of synthetic pyrethroid insecticide, deltamethrin (DM), can elicit pathophysiological effects through oxidative stress in non-targeted organisms such as mammals. There is accumulating evidence that virgin olive oil (VOO), a rich source of polyphenolic components, have anti-oxidant, anti-inflammatory, and anti-apoptotic properties. This study aimed to determine the protective and ameliorative effects of VOO against DM-induced nephrotoxicity. Methods & materials: Mice were randomly divided into four equal groups: DM group, DM plus VOO group, VOO group, and vehicle group. Five weeks after gavaging, kidney samples were taken for biochemical assessment of malondialdehyde (MDA), glutathione (GSH) and catalase (CAT), and for immunohistochemical assessment of caspase-3, cyclooxygenase-2 (cox-2) and poly (ADP-ribose) polymerase (PARP). Results: The MDA level in kidney was increased in the DM group, which was significantly decreased after VOO administration in the DM plus VOO group. The GSH level and CAT activiy in kidney were decreased in the DM group, which were significantly increased after VOO administration in the DM plus VOO group. Greater expression of caspase-3, cox-2, and PARP could be detected in the DM group, which was significantly attenuated in the DM plus VOO group. Also, the histopathological changes which were detected in the DM group attenuated after VOO consumption. Conclusion: Virgin olive oil exerted protective effects against deltamethrin-induced nephrotoxicity, which might be associated with its anti-apoptotic, anti-inflammatory, and anti-oxidative properties. Keywords: Deltamethrin, Virgin olive oil, Antioxidant, Apoptosis, Inflammation, Nephrotoxicit

Protective Effect of Atorvastatin on Benzopyrene-Induced Hepatorenal Toxicity: A Histopathological Study

Background and purpose: Benzo[a]pyrene (BaP) is an environmental pollutant that has genotoxic and carcinogenic effects. Atorvastatin (ATV), as a lipid-lowering drug, has antioxidant, anti-inflammatory and anti-apoptotic properties. This study investigated the effects of ATV on chronic liver and kidney damage induced by BaP. Materials and methods: In this experimental study, adult female rats were randomly divided into seven groups (n= 8 per group), including control group, olive oil group, ATV group (10 mg/kg), BaP groups (10 and 20 mg/kg), and ATV + BaP groups (10 and 20 mg). The drugs were administered by oral gavage for ten consecutive days. Histopathologic evaluation of the liver and kidney tissues was done one month after drug administration. Results: Histopathologic changes in both liver and kidney tissues such as inflammatory cell infiltration, plasma leakage, reduced acidophilia of hepatocytes, and renal tubular cells were seen in the groups receiving BaP. Findings showed that BaP at 20 mg/kg caused more serious harm than that at 10 mg/kg. ATV treatment protected structural changes in liver and kidney tissues. Conclusion: Current study showed that destructive effects of benzopyrene remain until one month after administration. The protective effects of atorvastatin against benzopyrene-induced hepatotoxicity were confirmed over time

Alleviation of cisplatin‐induced hepatotoxicity by gliclazide: Involvement of oxidative stress and caspase‐3 activity

Abstract Aims Cisplatin (CP), as an effective alkylating agent, is widely used in cancer treatment, while hepatotoxicity is one of its side effects. Gliclazide (GLZ), as an oral hypoglycemic drug, has antioxidant and anti‐inflammatory properties. This study was designed to investigate the protective effect of GLZ against CP‐induced hepatotoxicity in mice. Methods In this experimental study, 64 adult male mice randomly were allocated into eight groups (8 mice/group). Control, GLZ (5, 10, and 25 mg/kg, orally), CP (10 mg/kg, single dose, intraperitoneally), and CP+GLZ (in three doses). GLZ was administrated for 10 consecutive days. CP was injected on the 7th day of the study. At the end of the experiment, hepatotoxicity was evaluated by serum and tissue biochemical, histopathological, and immunohistochemical assessments. Results The data were revealed that CP increased oxidative stress (increased MDA and reduced GSH), liver damage enzymes (ALT, AST, and ALP), and immunoreactivity of caspase‐3 in liver tissue of CP‐injected mice. Also, CP induced histopathological changes such as eosinophilic of hepatocytes, dilatation of sinusoids, congestion, and proliferation of Kupffer cells. GLZ administration significantly ameliorated serum functional enzyme and hepatic oxidative stress markers in CP‐injected mice. In addition, the histological and immunohistochemical alterations were ameliorated in GLZ‐treated mice. Of the three doses, 10 and 25 mg/kg were more effective. Conclusions In conclusion, GLZ with its antioxidant, anti‐inflammatory, and anti‐apoptotic activities, can be suggested as a promising drug in the treatment of CP‐induced hepatotoxicity