Effect of transplantation of bone marrow stromal cell- conditioned medium on ovarian function, morphology and cell death in cyclophosphamide-treated rats

Objective: Although stem cell transplantation has beneficial effects on tissue regeneration, but there are still problems such as high cost and safety issues. Since stem cell therapy is largely dependent on paracrine activity, in this study, utilization of transplantation of bone marrow stromal cells [BMSCs]-secretome instead of the cells, into damaged ovaries was evaluated to overcome the limitations of stem cell transplantation

Materials and Methods: In this experimental study, BMSCs were cultured and 25-fold concentrated conditioned medium [CM] from BMSCs was prepared. Female rats were injected intraperitoneally with cyclophosphamide [CTX] for 14 days. Then, BMSCs and CM were individually transplanted into bilateral ovaries, and the ovaries were excised after four weeks of treatment. The follicle count was performed using hematoxylin and eosin [H and E] staining and the apoptotic cells were counted using TUNEL assay. Ovarian function was evaluated by monitoring the ability of ovulation and the levels of serum estradiol [E2] and follicle-stimulating hormone [FSH]

Results: Evaluation of the ovarian function and structure showed that results of secretome transplantation were almost similar to those of BMSCs transplantation and there was no significant differences between them

Conclusion: BMSCs-secretome is likely responsible for the therapeutic paracrine effect of BMSCs. Stem cell- secretome is expected to overcome the limitations of stem cell transplantation and become the basis of a novel therapy for ovarian damage

Supplementation of L-carnitine during in vitro maturation of mouse oocytes affects expression of genes involved in oocyte and embryo competence: an experimental study

Background: oocyte developmental competence is one of the key factors for determining the success rate of assisted reproductive technique

Objective: the aim of the current study was to investigate the effect of L-carnitine [LC] supplementation during in vitro maturation [IVM], on preimplantation embryo development and expression of genes involved in embryo competence derived from oocytes selected with brilliant cresyl blue [BCB] test

Materials and Methods: cumulus-oocyte complexes [COCs] were obtained from NMRI mice ovaries. COCs were stained with BCB and then BCB+ [colored cytoplasm] oocytes cultured in IVM medium supplemented with 0.3 or 0.6 mg/ml LC. COCs untreated with LC were used as control. Fertilization rate and blastocyst development rate were determined after in vitro fertilization. In addition, quantitative reverse transcriptase polymerase chain reaction was used to measure relative genes expression related with development [Ccnb1, Mos, Ces5, and Dppa2] and apoptosis [Bax and Bcl-xL] in oocytes and embryos

Results: oocytes treated with both LC concentrations showed higher blastocyst development rate compared with untreated oocytes [p<0.01]. Moreover, fertilization rate was increased in oocytes treated with 0.6 mg/ml LC [p<0.01]. Treatment of oocytes with both LC concentrations increased [p<0.01] the level of Ccnb1 mRNA in MII oocytes. The two-cell stage embryos and blastocysts derived from LC-treated oocytes [0.6 mg/ml] showed increased the expression levels of Dppa2 and Bcl-xl mRNA, respectively [p<0.01]

Conclusion: the results of the present study show that adding of LC to the IVM medium of BCB+ oocytes can ameliorate reproductive success following in vitro fertilization

Synthesis, and in-vitro cytotoxicity studies of a series of triazene derivatives on human cancer cell lines

Compounds containing triazene ring structure are cytotoxic agents and clinically used as antitumor alkylating agents. In this study, a series of triazene derivatives holding alkyl and aryl moieties were synthesized and proved to be potent cytotoxic agents in-vitro particularly against eight cancer cell lines [PCS, HT29, Hela, HL60, Jurkat, K562, MCF7, HepG2] and a non-cancerous cell line [HUVEC]. The cytotoxic activity was assessed using two methods, LDH assay, and trypan blue exclusion. Some of the triazene derivatives showed cytotoxic activity more than temozolomide [TMZ] as the reference drug. The synthesized triazenes showed marked cytotoxicity effects on all eight cancer cell lines. Among the compounds synthesized, l,3-bis [2-ethoxyphenyl] triazene C had unique efficacy and selectivity so that it had IC[50] between 0.560-3.33 microM on cancer cell lines and 12.61 microM on normal cell line [HUVEC]. l-[4-nitrophenyl]-3-[2-hydroxyethyl] triazene E shows weaker effect on cancer cell lines than the other compounds having 1C[50] between 3-15.54 microM

Prevention of di [2-ethylhexyl] phthalate-induced testicular disturbance in mice by Co-administration of L-carnitine

di [2-ethylhexyl] phthalate [DEHP] is widely used in the plastic industry and can induce reproductive toxicity. On the other hand, L-carnitine [LC] plays a crucial role in sperm metabolism and maturation. This study evaluates the effect of LC on body and testis weight, testis tissue, count, motility, viability, morphology, and chromatin quality of epididymal sperm, testicular spermatid number [TSN] per gram testis and daily sperm production [DSP] in LC-treated mice. In this experimental study, adult male NMRI mice [mean age: 4 weeks] were given doses of DEHP and LC by gavaging for 2 weeks. All samples were assessed according to World Health Organization [WHO] criteria. Sperm morphology was assessed using Papanicolaou staining and sperm chromatin quality by aniline-blue staining. The left testes were fixed in Bouin? solution for histological examination and the end slices were stained with hematoxylin and eosin [H and E]. The right testes were homogenized, and then TSN and DSP were calculated with an improved Neubauer haemocytometer and respective frames. Paired t-test, ANOVA, and Kruskal-Wallis tests were utilized for data analysis. Co-administration of DEHP and LC not only prevented significant gains in testicular weight, but also maintained the sperm's normal morphology and chromatin quality [p<0.05]. In addition, LC recovered histological changes, TSN, DSP, and sperm count. These results demonstrated that oral administration of LC partially or generally protects spermatogenesis from DEHP-toxicity in mice

[Histopathological study of Di-[2-ethylhexyl] phthalate [DEHP] on testes in mouse]

Di-[2-ethylhexyl] phthalate [DEHP] is plasticizer used commonly in a variety of polyvinyl chloride- based consumable products. The purpose of this study was to evaluate the effects of DEHP on body weight, testis weight, seminiferous tubular diameter, seminiferous epithelium height, seminiferous lumen diameter, number of sertoli cells and round spermatids per seminiferous tubule in mice. The protocol for DEHP administration was that adult male NMRI mice [the age group of 4 weeks] received 2g DEHP/100 micro l corn oil/kg, and vehicle group received 100 micro l corn oil/ kg by gavage for 14 days. The control group did not receive DEHP. All animals were weighed on the first and terminal day of the experiment. The left and right without fat testis, weights were recorded for each animal. Left testis was fixed in Bouins solution, routinely processed for embedding in paraffin and staining of 5 micro m sections with hematoxilin and eosin [H and E] for histopathological examination. Administration of DEHP induced the reduction of body and testis weight significantly [p<0.05]. Furthermore, DEHP decreased the seminiferous tubular diameter, seminiferous epithelium height, and seminiferous lumen diameter. The number of sertoli cells and round spermatids in seminiferous tubule was significantly low, compared with control group. These results demonstrated that DEHP administration has toxicant effects on body and testis weight, spermatogenesis process, along with male reproductive system