Melatonin improves the structure and function of autografted mice ovaries through reducing inflammation: A stereological and biochemical analysis.

Melatonin has anti-oxidant, anti-inflammatory and anti-apoptotic properties. We aimed to investigate the effect of melatonin on the structure and function of mice ovaries following autograft transplantation. NMRI mice were divided into: control, autografted + saline, autografted + melatonin (20 mg/kg/day i.p. injection for 1 day before until 7 days after transplantation). 28 days post transplantation, ovary compartments were studied stereologically. Follicle apoptosis and the level of progesterone and estradiol were also measured. The inflammation, serum MDA concentration and total antioxidant capacity were also assessed on day 7 post transplantation. The total volume of the ovary, cortex and medulla (P < 0.05) and the number of different types of follicles (P < 0.001), the concentration of IL-10, progesterone and estradiol (P < 0.001) and TAC (P < 0.01) significantly decreased in the autografted + saline group compared to the control. The levels of IL-6 (P < 0.01), TNF-α, MDA and the apoptotic rate (P < 0.001) increased significantly in the autografted + saline group compared to the control, while the total volume of the ovary, cortex and medulla (P < 0.05) and the number of different types of follicles (P < 0.001), the concentration of IL-10, progesterone and estradiol (P < 0.001) and TAC (P < 0.01) significantly increased in the autografted + melatonin group compared to the autografted + saline group. The levels of IL-6 (P < 0.01), TNF-α, MDA and the apoptotic rate (P < 0.001) decreased significantly in the autografted + melatonine group compared to the autografted + saline group. In the autografted + melatonin group, the localization of CD31-positive cells in the theca layer was similar to the control group. Melatonin can improve the structure and function of the grafted ovary.

Utilizing platelet-rich fibrin bioscaffold at the graft site improves the structure and function of mice ovarian grafts.

Aim: The effect of platelet-rich fibrin (PRF) bioscaffold on the structure and function of mice-autotransplanted ovaries was investigated. Materials & methods: Mice were divided into three groups: control, autografted and autografted + PRF bioscaffold. Angiogenesis, ovary histology and serum biochemical factors were assessed. Results: The total volume of the ovary, the number of follicles and the level of superoxide dismutase activity, total antioxidant capacity, IL-10, progesterone and estradiol were significantly higher in the autografted + PRF bioscaffold group compared with the autografted group. In the autografted + PRF bioscaffold group, angiogenesis was accelerated and apoptosis rate, IL-6, TNF-α, malondialdehyde concentrations were significantly lower compared with the autografted group. Conclusion: PRF bioscaffold improves the structure and function of mice-autografted ovary.

Adipose-derived mesenchymal stromal cell transplantation at the graft site improves the structure and function of autografted mice ovaries: a stereological and biochemical analysis.

BACKGROUND: Ovarian tissue autografting is a fertility restoration technique that is frequently used in young women with cancer who undergo radio/chemotherapy. A limiting factor in this technique is ischemia-reperfusion (I/R) damage. Because adipose-derived mesenchymal stromal cells (ADMSCs) protect different ischemic tissues against I/R damage, we examined the effect of ADMSC transplantation at the graft site in mice ovary autografting. METHOD: Mice were divided into three groups: control, autograft and autograft + ADMSCs. Seven days after ovary autografting and ADMSC transplantation, serum superoxide dismutase (SOD) activity, total antioxidant capacity, serum concentrations of malondialdehyde (MDA), tumor necrosis factor alpha (TNFα), interleukin (IL)-6 and IL-10 were measured. After 28 days, ovary histology, serum concentrations of progesterone and estradiol and apoptosis rate were also estimated. At 1-3 and 28 days post-ovary autografting and ADMSC transplantation, angiogenesis was detected. The results were analyzed using one-way analysis of variance (ANOVA) and Tukey test, and the means were significantly different at P ≤ 0.05. RESULT: In the autograft + ADMSCs group, the total volume of the ovary, cortex and medulla (P ≤ 0.001), the number of follicles, SOD activity, IL-10 (P ≤ 0.001) and progesterone and estradiol (P ≤ 0.01) concentrations significantly increased compared with the autograft group. Apoptosis rate, IL-6, TNFα and MDA concentrations in the autograft + ADMSCs group were lower than the autograft group (P ≤ 0.001). The angiogenesis was accelerated and the localization of CD31-positive cells in the cortex was similar to the control group following ADMSC transplantation. DISCUSSION: ADMSC transplantation enhances the structure and function of grafted ovary.

Biochemical and morphological changes in bone marrow mesenchymal stem cells induced by treatment of rats with p-Nonylphenol.

OBJECTIVES: In previous investigations, we have shown para-nonylphenol (p-NP) caused significant reduction of proliferation and differentiation of rat bone marrow mesenchymal stem cells (MSCs) in vitro. In this study, we first treat the rats with p-NP, then carried out the biochemical and morphological studies on MSCs. MATERIALS AND METHODS: Proliferation property of cells was evaluated with the help of MTT assay, trypan blue, population doubling number, and colony forming assay. Differentiation property was evaluated with quantitative alizarin red assay, measurement of alkaline phosphatase (ALP) activity as well as intracellular calcium content. In addition; morphological study, TUNEL test, activated caspase assay, and comet assay were performed to evaluate the mechanism of the cell death. RESULTS: The results showed significant reduction in the colony-forming-ability and population-doubling-number of extracted cells when compared to control ones. In addition, it was revealed that the p-NP treatment of rats caused significant reduction in nuclear diameter, cytoplasm shrinkage, and induction of caspase-dependent-apoptosis. Also there was significant reduction in ALP activity, intracellular calcium content, and intracellular matrix following osteogenic differentiation. CONCLUSION: As MSCs are the cellular back up for bone remodeling and repair, we suggest more investigations to be conducted regarding the correlation between the increasing number of patients suffering from osteoporosis and p-NP toxicity. Also, we strongly recommend WHO and local health organization to prevent industries of using p-NP in formulation of industrial products which may cause changes in proliferation and differentiation properties of stem cells.

Induction of Apoptosis in the Rat Bone Marrow Mesenchymal Stem Cells Following Sodium Arsenite Treatment with the Dose Lesser than that Used for Treatment of Malignant Patient.

OBJECTIVE(S): Arsenic compounds are potent human carcinogen and produce a variety of stress responses in mammalian cells. Recently sodium arsenite has been recommended to be used as anti malignancy drug by American food and drug administration (FDA). In this study, we aimed to determine the apoptosis inducing effect of sodium arsenite on rat bone marrow mesenchymal stem cells exposed in vitro. METHODOLOGY: Cell morphology was studied with the help of Hoechst and propidium iodide as well as with single cell gel electrophoresis(comet assay), TUNEL assay and caspase activity base on immunocytochemistry using commercial kit were considered to study the mechanism of cell death. RESULTS: Our result showed that the sodium arsenite with concentration of 0.1 µM in 36 hr induces caspase dependent apoptosis in rat bone marrow mesenchymal stem cells. This concentration is the lowest level of sodium arsenite to be reported with apoptosis induction ability in stem cells. CONCLUSION: Since sodium arsenite is used in therapy, more research should be carried out on the effect of this chemical on stem cells, especially MSCs.