Fortified electrospun collagen utilizing biocompatible Poly Glycerol Sebacate prepolymer (PGSp) and zink oxide nanoparticles (ZnO NPs) for diabetics wound healing: Physical, biological and animal studies.

Collagen, a naturally occurring fibrous protein, is a potential resource of biological materials for tissue engineering and regenerative medicine because it is structurally biocompatible, has low immunogenicity, is biodegradable, and is biomimetic. Numerous studies have documented in the literature how Collagen nanofibers exhibit limited cell adhesion, poor viscosity, and no interior fibril structure. The biomedical industry is using Poly Glycerol Sebacate prepolymer(PGSp), a biodegradable and biocompatible polyester with high adhesion and very viscous appearance, more often. Here, unique electrospun Collagen/PGSp/ZnO/NPs blend nanofibers for skin tissue application were developed and described with varied PGSp percent. Additionally, when ternary blends of Collagen, PGSp, and Zink Oxide Nanoparticles (ZnO NPs) are used, the antibacterial properties of the scaffolds are improved. The bead-free electrospun nanofibers were produced by raising the PGSp concentration to 30%w/w. SEM, EDS, tensile, MTT, FTIR, SDS-page, swelling test, contact-angle, antimicrobial, biodegradation, XRD, and cell attachment procedures were used to characterize the crosslinked nanofibers. The ternary blend nanofibers with a weight ratio of Collagen/PGSp 30%/ZnONPs 1% had higher stress/strain strength (0.25 mm/mm), porosity (563), cell survival, and degradation time. Moreover, after applying for wound healing in diabetic rats, Collagen/PGSp 30%/could be show improving wound healing significantly compared to other groups.

Role of Menstrual Blood Stem Cell-Derived Secretome, Follicular Fluid, and Melatonin in Oocyte Maturation and Embryo Development in Polycystic Ovary Syndrome.

BACKGROUND: In vitro maturation has been considered an approach to mature oocytes derived from women with polycystic ovary syndrome (PCOS). It is suggested that the IVM of oocytes may benefit from mesenchymal stem cells derived conditioned medium (CM-MSC). OBJECTIVE: The purpose of this study was to determine the efficacy of a cocktail of menstrual blood stem cell (MenSCs)-derived secretome, along with follicular fluid and melatonin, in oocyte maturation and embryo development in PCOS. METHODS: Four hundred left germinal vesicle oocytes were collected from 100 PCOS patients and randomly divided into four treatment groups: 1) control, 2) secretome, 3) follicular fluid, and 4) melatonin. Oocyte maturation, fertilization rate, and embryo development were monitored, as well as the expression levels of oocyte-secreted factors (GDF9- BMP15), oocyte maturation (MPK3), and apoptosis (BAX- Bcl2). RESULTS: The rate of oocyte maturation increased in all test groups, but only the results for the SEC group were significant (P= 0.032). There were no significant differences in oocyte fertilization and embryo yield among groups. However, the quality of embryos significantly increased in the melatonin group compared to the control. Cytoplasmic maturation was confirmed by the expression of oocyte maturation-related genes using Real-time PCR. Additionally, the expression level of BCL-2 was significantly higher in the SEC-FF-MEL group than in the control group (p ≤ 0.01). CONCLUSION: Enrichment of IVM media using MenSCs-secretome, particularly along with melatonin, could be an effective strategy to improve oocyte maturation and embryo development in PCOS.

Methylation Status of cAMP-responsive Element Modulator (CREM) Gene in Infertile Men and Its Association with Sperm Parameters.

The methylation pattern of non-imprinting genes was little studied, although it is widely known that the abnormal methylation levels of imprinting genes are associated with different forms of male infertility. The purpose of this research was to assess the CREM gene's methylation status and seminal characteristics in infertile individuals who were potential intracytoplasmic sperm injection (ICSI) candidates. A total of 45 semen samples (15 normospermia, 15 asthenospermia, and 15 oligoasthenoteratospermia) were examined. Using aniline blue (AB) staining, we carried out conventional semen analysis, chromatin quality, and sperm maturity testing. DNA was taken from semen samples, and all isolated DNA was assessed using Nanodrop and gel electrophoresis. A quantitative methylation-specific polymerase chain reaction (Q-MSP) approach was used to quantify the methylation at the DMRs of the CREM gene. According to our findings, sperm count (P=0.012), concentration (P= 0.019), motility (P=0.006), progression (P=0.006), and normal morphology (P=0.004) were all inversely correlated with abnormal sperm chromatin condensation. Additionally, we noted that the methylation level of the CREM gene was considerably more significant in the oligoasthenoteratospermia group compared to the asthenospermia and normospermia groups (P<0.05). Additionally, sperm count (P=0.043), progression (P=0.026), and normal morphology (P=0.024) were all inversely linked with CREM methylation. Overall, the abnormal CREM methylation patterns have a negative impact on sperm parameters. Additionally, the CREM gene's DNA methylation status may serve as an epigenetic indicator of male infertility.

Effect of Exosomes Derived from Bone Marrow Mesenchymal Stem Cells on Ovarian Granulosa Cells of Immature NMRI Mice.

OBJECTIVE: In recent years, in vitro maturation (IVM) has become the focus of fertility maintenance, and infertility treatment. The aim of this study is development of oocytes during folliculogenesis and oogenesis is greatly influenced by the presence of BMP-7, BMP-15, and GDF-9 genes, which are present in exosomes generated from bone marrow stem cells. MATERIALS AND METHODS: In the experimental study, we investigated how exosomes obtained from bone marrow stem cells affected development and expansion of ovarian granulosa cells (GCs) in NMRI mice. In this in vitro experiment, bone marrow stem cells were isolated from mice's bone marrow, and after identification, exosomes were recovered. Exosome doses of 100, 50, and 25 µg/ml were applied to GCs before using MTT assay to measure survival rates and quantitative reverse-transcription polymerase chain reaction (PCR) to measure expression of the BMP-7, BMP-15, and GDF-9 genes. RESULTS: The results showed that the GCs treated with exosomes concentrations of 25, 50, and 100 µg/ml significantly increased bioavailability, growth and proliferation and it also increased expression level of BMP-7, BMP-15, and GDF-9 genes compared to the controls. CONCLUSION: Findings of this study indicated that exosomes derived from bone marrow stem cells improved growth of GCs in NMRI mice and they were a good candidate for further clinical studies to improve quality of the assisted reproductive techniques.

In vitro pro-apoptotic and anti-metastatic potentials of harmaline-silver containing folate-linked chitosan nano-drug delivery system.

Objectives: Harmaline and green-synthesized silver nanoparticles were encapsulated by folate-linked chitosan molecules as a receptor-mediated drug delivery system to evaluate its pro-apoptotic and anti-metastatic potentials on human ovarian (A2780) and epithelioid (PANC) cancer cells. Materials and Methods: The Ag nanoparticles (AgNP) were synthesized utilizing an herbal bio-platform (Bistorta officinalis) and embedded with harmalin. The Harmaline-ag containing folate-linked chitosan nanoparticles (HA-fCNP) were synthesized utilizing the ionic gelation method. Both the AgNP and HA-fCNP nanoparticles were characterized by DLS, FESEM, and Zeta potential analysis. Moreover, the chemical properties of HA-fCNP and the crystallinity of AgNPs were determined by applying FTIR and XRD methods, respectively. The HA-fCNP cytotoxicity was analyzed on A2780, PANC, and HFF cell lines. Moreover, pro-apoptotic and anti-metastatic potentials of HA-fCNP were studied by analyzing the BAX-BCL2 and MMP2-MMP9 gene expression profiles, respectively. The A2780 cellular death was determined by AO/PI and flow cytometry methods. Results: The HA-fCNP significantly exhibited a selective cytotoxic impact on A2780 and PANC cancerous cell lines compared with normal human foreskin fibroblast (HFF) cells. The increased SubG1-arrested A2780 cells and up-regulated BAX gene expression following the increased treatment concentrations of hA-fCNP indicated its selective pro-apoptotic activity on A2780 cells. Also, the notable down-regulated expressions of MMP2 and MMP9 metastatic genes following the increasing doses of HA-fCNP treatment on A2780 cells confirmed its anti-metastatic activity. Conclusion: The cancer-selective cytotoxicity, apoptotic, and anti-metastatic properties of HA-fCNP are considered the appropriate properties of an anticancer compound.

Protective Effect of Docetaxel Against Autophagy-Related Genes in Vitrification of Mouse Metaphase II Oocytes.

Background: Autophagy is a conservative mechanism for cell survival as the main response of cells to stress conditions. The present study aimed to assess the effect of docetaxel on the survival, fertilization, and expression of autophagy-related genes in vitrified oocytes. Methods: The study was conducted in 2018 at the Stem Cells Technology Research Center, Shiraz University of Medical Sciences (Shiraz, Iran). Denuded oocytes were randomly selected and assigned to five groups, namely control (n=133), docetaxel (n=136), docetaxel+cryoprotectants (n=146), docetaxel+vitrification (n=138), and vitrification (n=145). The effect of vitrification on the expression of autophagy-related gene 5 (ATG5) and Beclin-1 was determined using a real-time polymerase chain reaction. Data were analyzed using SPSS software (version 26.0) and GraphPad Prism 9. Results: Survival and fertilization rates in each experimental group were significantly reduced compared to the control group (P=0.001). After in vitro fertilization of oocytes, the 2-cell formation rate was significantly reduced in the docetaxel+vitrification and vitrification groups compared to the control and docetaxel groups (P=0.001 and P=0.001, respectively). Pre-incubation of oocytes with docetaxel reduced gene expression levels of Beclin-1 and ATG5 in the docetaxel+cryoprotectants and docetaxel+vitrification groups (P=0.001 and P=0.019, respectively). The expression level of these genes was also reduced in the docetaxel group compared to the control group (P=0.001). Conclusion: Incubation of mouse metaphase II oocytes with docetaxel prior to vitrification reduced the expression of autophagy-related genes and increased survival and fertilization rates compared to untreated oocytes.

Association of Cytochrome P450 2E1 and N-Acetyltransferase 2 Genotypes with Serum Isoniazid Level and Anti-Tuberculosis Drug-Induced Hepatotoxicity: A Cross-Sectional Study.

Background: Anti-tuberculosis drug-induced hepatotoxicity can result from genetic polymorphism of the isoniazid (INH) metabolizing enzyme. This study aimed to determine the effect of genetic polymorphism of N-acetyltransferase 2 (NAT2) and cytochrome P450 2E1 (CYP2E1) genes on serum isoniazid level and drug-induced hepatotoxicity. Methods: A cross-sectional study was conducted on 120 patients (with and without hepatotoxicity) with pulmonary tuberculosis from June 2019 to April 2022 in Tehran (Iran). High-performance liquid chromatography was used to measure the serum concentration of INH and acetylisoniazid (AcINH). NAT2 and CYP2E1 genotypes were determined using polymerase chain reaction and restriction fragment length polymorphism methods. Data were analyzed using SPSS software (version 22.0) with independent two-sample t test, Chi square test, or Fisher's exact test. P<0.05 was considered statistically significant. Results: A total of 40 patients showed hepatotoxicity. The risk of anti-tuberculosis drug-induced hepatotoxicity was significantly higher in patients who are slow acetylator (SA) phenotype than in rapid or intermediate acetylator (P<0.001). NAT2*4/*4 genotypes were not found in patients with hepatotoxicity. The frequency of NAT2*5 and NAT2*6 haplotypes and serum INH concentration was significantly higher in patients with hepatotoxicity than in those without (P=0.003, P<0.001, and P<0.001, respectively). NAT2*4 haplotype was correlated with protection against hepatotoxicity. A combination of SA and CYP2E1 C1/C1 genotype was significantly associated with hepatotoxicity (P<0.001). Conclusion: Hepatotoxicity in Iranian patients with tuberculosis was confirmed due to the presence of NAT2 SA polymorphism. Determining NAT2 and CYP2E1 genotypes and/or INH concentration can be a valuable tool to identify patients susceptible to hepatotoxicity.

Treatment with quercetin increases Nrf2 expression and neuronal differentiation of sub ventricular zone derived neural progenitor stem cells in adult rats.

BACKGROUND: The presence of neural precursor stem cells (NPSCs) in some parts of the adult brain and the potency of these types of cells with a therapeutic viewpoint, has opened up a new approach for the treatment and recovery of the defects of central nervous system (CNS). Quercetin, as an herbal flavonoid, has been extensively investigated and shown to have numerous restoratives, inhibitory, and protective effects on some cell-lines and disorders. The purpose of this study is to simultaneously investigate the effect of quercetin on the expression of the nuclear factor erythroid 2-related factor 2 (Nrf2) gene and the effect on the proliferation and differentiation of NPSCs derived from the subventricular zone (SVZ) of the brain of adult rats. METHODS AND RESULTS: The cell obtained from SVZ cultured for one week and treated with quercetin at the concentrations of 1, 5, and 15 µM to evaluate the Nrf2 expression, proliferation and differentiation of NSCs after one week. Cellular and genetic results was performed by RT-PCR, MTT assay test, quantification of images with Image-J and counting. The results indicated that the quercetin increases expression of Nrf2 at concentration above 5 µM. Also differentiation and proliferation rate of NSCs is affected by various concentrations of quercetin in a dose-dependent manner. CONCLUSION: These findings confirmed the dose-dependent effect of quercetin on proliferation and differentiation of cell. In addition, quercetin increased the expression of Nrf2 gene. By combining these two effects of quercetin, this substance can be considered an effective compound in the treatment of degenerative defects in CNS.

Prenatal kisspeptin antagonist exposure prevents polycystic ovary syndrome development in prenatally-androgenized rats in adulthood: An experimental study.

Background: Increased levels of kisspeptin are associated with hypothalamus-pituitary-ovary axis dysfunction. It may lead to the development of polycystic ovary syndrome (PCOS). Objective: We aimed to investigate the effect of prenatal kisspeptin antagonist exposure on the development of PCOS in prenatally androgenized rats in adulthood. Materials and Methods: In this experimental study, pregnant rats were injected with free testosterone (T, 5 mg/day) or T+P271 (kisspeptin antagonist) on the 20 th day of the pregnancy period (n = 5 in each group), while rats in the control group received solvent. Female offspring were examined in terms of anogenital distance (AGD), anovaginal distance (AVD), vaginal opening, serum total testosterone (TT) levels, ovarian follicles, and the regularity of estrous cycles in adulthood. AGD and AVD were measured using a vernier caliper. TT levels were measured using the enzyme-linked immunosorbent assay method. Ovaries were fixed in 10% formalin, tissue processing was done by a standard protocol, and then ovaries embedded in paraffin. 5 µm-thickness ovarian sections mounted on a glass slide, deparaffinized, and stained using Harris's Hematoxylin and Eosin Y. Results: AGD, AVD (p < 0.001), TT levels (p = 0.02), and the numbers of preantral and antral follicles (p < 0.001) in the ovaries were significantly decreased in prenatally T-P271-exposed rats compared to prenatally T-exposed rats. The age of vaginal opening was early in T-P271-exposed rats compared to prenatally T-exposed rats (p < 0.001). The number of corpora lutea was significantly increased in T-P271-exposed rats (p < 0.001). No cystic follicles were observed in the ovaries of prenatally T-P271-exposed rats. Prenatally T-P271-exposed rats had regular estrous cycles compared to prenatally T-exposed rats. Conclusion: Prenatal exposure to kisspeptin antagonist can prevent PCOS development in prenatally androgenized rats in adulthood.

Effects of empagliflozin on the expression of kisspeptin gene and reproductive system function in streptozotocin-induced diabetic male rats.

One of the main health concerns of diabetes is testicular dysfunction and impairment of reproductive function and sperm quality which can cause male infertility. kisspeptin is a hypothalamic neuropeptide hormone that is involved in the regulation of energy metabolism, gonadotrophin-releasing hormone (GnRH), and reproductive function. In the present study, the therapeutic effects of empagliflozin (sodium-glucose co-transporter 2 inhibitors) on kisspeptin expression along with reproductive function were investigated in diabetic male Wistar rats. Diabetes was induced by a single dose injection of 60 mg/kg streptozotocin. Empagliflozin in doses of 10 and 25 mg/kg body weight was used for 8 weeks. Serum samples, testis, epididymis, and pancreas tissues were collected at the end of the experiments. Lipid profiles, oxidative stress markers, blood hormones, expression of kisspeptin along with pathological alterations of the testis were assayed using real-time PCR, biochemical, and histological technics. Data have shown that empagliflozin improved hyperglycemia, reproductive impairment, oxidative stress condition, and histopathological alterations of pancreatic and testis tissues in diabetic animals. It improved the serum levels of sex hormones, insulin, leptin, and the expression of kisspeptin in the testes tissues. Spermatogenesis is also improved in treated animals. Data indicated that the administration of empagliflozin can ameliorate symptoms of diabetes. It probably has promising antidiabetic potential and may improve the male infertility of diabetic subjects. To our knowledge, this is the first experimental evidence for the potential impact of empagliflozin on kisspeptin expression in diabetic male rats.

Astaxanthin ameliorates the impairment consequence of prenatal bacterial lipopolysaccharide exposure in adult male offspring NMRI mice.

In this study, the potential effects of astaxanthin (AST) were investigated on preventing the prenatal LPS-induced injures in mothers and adult male offspring of NMRI mice. Pregnant mice were randomly divided into four groups: 1. Saline + vehicle; 2. Saline + AST: received astaxanthin (4 mg/kg for 3 days, ip) on 11-13 gestation days; 3. LPS + vehicle (LPS-treated group): injected with LPS (20 µg/kg, sc) on gestation day 11; 4. LPS + AST: administrated LPS and astaxanthin on gestation days 11 and 11-13, respectively. In each group, maternal care behaviors and TNF-α serum levels were examined until weaning of male offspring at 23 days. At 60 days old, male pups underwent analysis of body weight and length, serum gonadotropins and testosterone hormone levels, sperm quality, gonadal and brain tissues morphologies, and the expression of SOX9 and GnRH genes by real-time PCR. Serum TNF-α level increased significantly in mothers treated with LPS, while AST reduced it. In adult male offspring, serum hormone levels, sperm quality, and the number of spermatocytes and Leydig cells in the testes improved when AST was administrated. According to histological studies of the brain, neurons in the LPS-treated group were smaller and less active, whereas neurons in the LPS + AST group were larger, more numerous, and more active. LPS significantly reduced GnRH expression, while AST induction improved its expression. AST administration during pregnancy prevented the adverse effects of prenatal exposure to LPS, presumably through its genomic and non-genomic effects, in adult male offspring.

Differentiation of Alginate-Encapsulated Wharton Jelly-Derived Mesenchymal Stem Cells into Insulin Producing Cells.

Objective: Insulin insufficiency due to the reduced pancreatic beta cell number is the hallmark of diabetes, resulting in
an intense focus on the development of beta-cell replacement options. One approach to overcome the problem is to
search for alternative sources to induce insulin-producing cells (IPCs), the advent of mesenchymal stem cells (MSCs)
holds great promise for producing ample IPCs. Encapsulate the MSCs with alginate improved anti-inflammatory effects
of MSC treatment. This study aimed to evaluate the differentiation of wharton jelly-derived MScs into insulin producing
cells using alginate encapsulation.
Materials and Methods: In this experimental study, we established an efficient IPCs differentiation strategy of human
MSCs derived from the umbilical cord's Wharton jelly with lentiviral transduction of Pancreas/duodenum homeobox
protein 1 (PDX1) in a 21-day period using alginate encapsulation by poly-L-lysine (PLL) and poly-L-ornithine (PLO)
outer layer. During differentiation, the expression level of PDX1 and secretion of insulin proteins were increased.
Results: Results showed that during time, the cell viability remained high at 87% at day 7. After 21 days, the differentiated beta-like cells in microcapsules were morphologically similar to primary beta cells. Evaluation of the expression of PDX1 and INS by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on days 7, 14 and 21 of differentiation exhibited the highest expression on day 14. At the protein level, the expression of these two pancreatic markers was observed after PDX1 transduction. Results showed that the intracellular and extracellular insulin levels in the cells receiving PDX1 is higher than the control group. The current data showed that encapsulation with alginate by PLL and PLO outer layer permitted to increase the microcapsules' beta cell differentiation.
Conclusion: Encapsulate the transduced-MSCs with alginate can be applied in an in vivo model in order to do the further analysis.

Local Insulin-Derived Amyloidosis Model Confronted with Silymarin: Histological Insights and Gene Expression of MMP, TNF-α, and IL-6.

Amyloidosis is a heterogeneous group of protein deposition diseases associated with the presence of amyloid fibrils in tissues. Analogs of insulin that are used for treating diabetic patients (including regular insulin) can form amyloid fibrils, both in vitro and in vivo as reported in patients. The main purpose of this study was the induction of localized insulin-generated amyloidosis and the observation of silymarin effects on this process. In order to obtain amyloid structures, regular insulin was incubated at 37 °C for 24 h. Congo red absorbance and transmission electron microscopy images validated the formation of amyloid fibrils. Those fibrils were then injected subcutaneously into rats once per day for 6, 12 or 18 consecutive days in the presence or absence of silymarin, and caused development of firm waxy masses. These masses were excised and stained with Hematoxylin and Eosin, Congo red and Thioflavin S. Histological examination showed adipose cells and connective tissue in which amyloid deposition was visible. Amyloids decreased in the presence of silymarin, and the same effect was observed when silymarin was added to normal insulin and injected subsequently. Furthermore, plasma concentrations of MMP2, TNF-α, and IL-6 inflammatory factors were measured, and their gene expression was locally assessed in the masses by immunohistochemistry. All three factors increased in the amyloidosis state, while silymarin had an attenuating effect on their plasma levels and gene expression. In conclusion, we believe that silymarin could be effective in counteracting insulin-generated local amyloidosis.

Therapeutic Effect of P-Cymene on Lipid Profile, Liver Enzyme, and Akt/Mtor Pathway in Streptozotocin-Induced Diabetes Mellitus in Wistar Rats.

Diabetes is a serious public health problem in low- and middle-income countries. There is a strong link between hyperglycemia, oxidative stress, inflammation, and the development of diabetes mellitus. PI3K/Akt/mTOR is the main signaling pathway of insulin for controlling lipid and glucose metabolism. P-cymene is an aromatic monoterpene with a widespread range of therapeutic properties including antioxidant and anti-inflammatory activity. In the present study, the antidiabetic effects of p-cymene were investigated. Diabetes was induced using streptozotocin in male Wistar rats. The effects of p-cymene and metformin were studied on levels of glucose (Glu), lipid profile, liver enzymes, oxidative stress, and the expression of Akt, phospho-Akt, and mTOR (mammalian target of rapamycin) proteins, using biochemical, histological, and immunohistochemical analysis. Data have shown that p-cymene can improve serum levels of Glu, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and the expression of mTOR, Akt, and phospho-Akt protein in diabetic animals. These results suggest that p-cymene has hypoglycemia, hypolipidemia, and antioxidant properties. It can regulate Akt/mTOR pathway and reduce hepatic and pancreas injury. It can be suggested for diabetes management alone or simultaneously with metformin.

Novel electrospun conduit based on polyurethane/collagen enhanced by nanobioglass for peripheral nerve tissue engineering.

Peripheral nerve injury can significantly affect the daily life of individuals with impaired nerve function and permanent nerve deformity. One of the most common treatments is autograft transplantation. Tissue engineering is one of the efficient methods to regenerate injured nerves using scaffolds, cells, and growth factors. Conduits, which are produced by a variety of techniques, could be used as an alternative treatment for patients with damaged nerves. The electrospinning technique is one of the most important and widely used methods for generating nanofiber conduits from biocompatible polymers. In this study, using the electrospinning method, three different conduits, including polyurethane (PU), polyurethane/collagen (PU/C), and a new conduit based on polyurethane + collagen + nanobioglass (PU/C/NBG), were prepared. The characteristics of these three types of conduits were evaluated by SEM, XRD, and various experiments, including porosity, degradation, contact angle, DMTA, FTIR, MTT, and DAPI staining. The results of MTT and DAPI assays revealed the safety of conduits and proper cell attachment. Overall, the results obtained from various experiments showed that the novel PU/C/NBG conduit has better mechanical properties in terms of porosity, hydrophilicity, and biocompatibility in comparison with PU and PU/C conduits and could be a suitable candidate for peripheral nerve regeneration and axonal growth due to its repair potential.

Protective Effect of Cerium Oxide Nanoparticles on Human Sperm Function During Cryopreservation.

The generation of reactive oxygen species during cryopreservation of human sperm has negative effects on the consistency of the thawed sperm. The antioxidant properties of cerium oxide nanoparticles (CeO2NPs) may be useful for reducing cryodamage in thawed sperm. This research was conducted to determine the effects of CeO2NPs on the quality and function of human sperm after thawing. Samples of semen obtained from 20 normozoospermic individuals were allocated to the following four groups: fresh, frozen control (sperm not treated with CeO2NPs), and those exposed to 0.1 µg/mL CeO2NPs (CeO2-0.1), 1 µg/mL CeO2NPs (CeO2-1), and 5 µg/mL CeO2NPs (CeO2-5). Sperm parameters of motility, viability, membrane integrity, DNA fragmentation, protamination, malondialdehyde (MDA) levels, mitochondria membrane potential, and morphology were evaluated after the freezing-thawing process. The results showed that 0.1 µg/mL CeO2NPs significantly (p < 0.05) improved the following human sperm parameters after thawing: progressive (44.6% ± 1.14% vs. 36.2% ± 1.24%) and total motility (60.9% ± 2.5% vs. 51.3% ± 2.5%), viability (67.9% ± 1.5% vs. 58.1% ± 1.5%), membrane functionality (66.1% ± 1.85% vs. 55.4% ± 1.85%), DNA integrity (30.8% vs. 24.04%), and protamination (69.85% ± 2.09% vs. 57.2% ± 2.09%) compared with the frozen control group. We observed the lowest MDA levels in the CeO2-0.1 (3.06 ± 0.25 nmol/mL), CeO2-1 (3.1 ± 0.25 nmol/mL), and CeO2-5 (3.08 ± 0.25 nmol/mL) groups compared with the frozen control group (3.72 ± 0.25). Different concentrations of CeO2NPs did not significantly change sperm normal morphology and mitochondria activity (p < 0.05).

Fabrication and characterization of biaxially electrospun collagen/alginate nanofibers, improved with Rhodotorula mucilaginosa sp. GUMS16 produced exopolysaccharides for wound healing applications.

Fabrication of scaffolds with enhanced mechanical properties and desirable cellular compatibility is critical for numerous tissue engineering applications. This study was aimed at fabrication and characterization of a nanofiber skin substitute composed of collagen (Col)/sodium alginate (SA)/ polyethylene oxide (PEO)/Rhodotorula mucilaginosa sp. GUMS16 produced exopolysaccharides (EPS) were prepared using the biaxial electrospinning technique. This study used collagen extracted from the bovine tendon as a natural scaffold, sodium alginate as an absorber of excess wound fluids, and GUMS16 produced exopolysaccharides as an antioxidant. Collagen was characterized using FTIR and EDS analyses. The cross-linked nanofibers were characterized by SEM, FTIR, tensile, contact-angle, swelling test, MTT, and cell attachment techniques. The average diameter of Col nanofiber was 910 ± 89 nm. The Col and Col-SA/PEO non-woven mats' water contact angle measurement was 41.6o and 56.4o, Col/EPS1%, Col/EPS2%, Col-SA/PEO + EPS1%, and Col-SA/PEO + EPS2% were 61.4o, 58.3o, 38.5o, and 50.6o, respectively. Cell viability of more than 100% was shown in Col-SA/PEO + EPS nanofibers. Also, SEM images of cells on nanofiber scaffolds demonstrated that all nanofibers incorporated with GUMS16-produced EPS have good cell growth and proliferation. The acquired results expressed that the GUMS16-produced EPS can be considered a novel biomacromolecule in electrospun fibers that increase cell viability and proliferation.

Nanobioglass enhanced polyurethane/collagen conduit in sciatic nerve regeneration.

The main purpose of neural tissue engineering and regenerative medicine is the development of biological substitutions to preserve, improve, and regenerate the damaged functions of tissues and organs. Three novel conduits, including polyurethane (PU), polyurethane/collagen (PU/C), and polyurethane/collagen/nano-bio glass (PU/C/NBG), were fabricated by the electrospinning technique. After confirming the suitability of conduits in the in-vitro environment, conduits were surgically sutured in a 10-mm gap in the sciatic nerve of a rat to evaluate their role in sciatic nerve reconstruction. After 4, 8, and 12 weeks of surgery, nerve regeneration was assessed by the hot plate test, sciatic functional index, electromyography, histology, and immunohistochemistry against S100, NF200, and CD31 antibodies. The results of various examinations revealed that the PU/C/NBG conduit is significantly more suitable than PU and PU/C conduits in terms of nerve regeneration. However, all three groups of conduits had the potential to be used for nerve regeneration. Overall, this study discovered that the PU/C/NBG conduit is a biocompatible neural conduit, which is a favorable candidate for peripheral nerve regeneration and axonal growth.

Nitric oxide could allay arsenic phytotoxicity in tomato (Solanum lycopersicum L.) by modulating photosynthetic pigments, phytochelatin metabolism, molecular redox status and arsenic sequestration.

Plants do not always have the genetic capacity to tolerate high levels of arsenic (As), which may not only arrest their growth but pose potential health risks through dietary bioaccumulation. Meanwhile, the interplay between the tomato plants and As-NO-driven molecular cell dynamics is obscure. Accordingly, seedlings were treated with As (10 mg/L) alone or in combination with 100 µM sodium nitroprusside (SNP, NO donor) and 200 µM 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, NO scavenger). Sodium nitroprusside immobilized As in the roots and reduced the shoot translocation by up-regulating the transcriptional expression of the PCS, GSH1, MT2, and ABC1. SNP further restored the growth retardation through modulating the chlorophyll and proline metabolism, increasing NO accumulation and stomatal conductance along with clear crosstalk between the antioxidant activity as well as glyoxalase I and II leading to endogenous H2O2 and MG reduction. Higher PCs and glutathione accumulation helped protect photosynthetic apparatus; however, cPTIO reversed the protective effects of SNP, confirming the role of NO in the As toxicity alleviation.

The Effect of N-Acetyl-Cysteine on NRF2 Antioxidant Gene Expression in Asthenoteratozoospermia Men: A Clinical Trial Study.

BACKGROUND: One of the important factor associated with male infertility is high production of reactive oxygen species (ROS). The main function of Nuclear factor erythroid 2-related factor 2 (NRF2) is to activate the cellular antioxidant response by inducing the transcription of a wide array of genes that can combat the harmful effects of factors such as oxidative stress. The purpose of this study was to evaluate the effect of N-acetyl-L-cysteine (NAC), as an antioxidant drug, on NRF2 Gene Expression in Asthenoteratozoospermia Men. MATERIALS AND METHODS: In this randomized, blinded clinical trial study, included 50 infertile men with asthenoteratozoospermia, who received NAC (600 mg, three times daily). Sperm parameters analyzed according to the world health organization (WHO; 2010). Sperm DNA fragmentation, relative NRF2 expression, and seminal plasma level of antioxidant enzymes were measured by TUNEL assay, reverse transcription polymerase chain reaction (RT-PCR) and ELISA test, respectively. RESULTS: After NAC treatment, findings showed a significant increase in sperm concentration and motility compared to pre-treatment status, whereas the percentage of abnormal morphology and DNA fragmentation was significantly decreased (P<0.05). A significant improvement in expression of NRF2 gene and antioxidant enzyme levels were observed compared to pre-treatment by NAC (P<0.05). Significant correlations were observed between NRF2 mRNA expression level, specific sperm parameters and level of antioxidant enzymes (P<0.05). CONCLUSION: The results demonstrated that NAC oral supplementation protected against oxidative stress by enhancing NRF2 expression. This could improve semen parameters quality parameters in asthenoteratozoospermia men (Registration number: IRCT20170830035998N4).