Bone Marrow Of Male Mice Research Articles

Evaluation of the genotoxicity of the food dye tartrazine in a micronucleus test in vivo

Introduction. Food azo dye Tartrazine (E102) is widely used in the production of food, pharmacological and cosmetic products. Despite the approval for use, interest in a comprehensive assessment of the impact of food colours, especially synthetic ones, on health continues unabated. The analysis of literature evaluating the genotoxicity of Tartrazine in vivo studies revealed some inconsistencies in the results, that showed the possibility to test a retail food colouring in one of recommended tests. Materials and methods. The Tartrazine genotoxicity (produced in India, purity 88.37%) was studied in the micronucleus test on male mouse bone marrow cells (hybrids F1 CBA x C57Bl6/j). The test substance was double enteral administrated in the dose range 250-2000 mg/kg. The frequency of polychromatophilic erythrocytes (PCEs) with micronuclei (MNs) was estimated by the analysis of 4000 PCEs. The proportion of PCE among all erythrocytes was determined by analyzing 1000 cells per animal. Results. There was no increase in the frequency of micronucleated polychromatophilic erythrocytes over a concurrent negative control with double enteral administration of Tartrazine in all doses studied. The proportion of PCEs among all erythrocytes did not change. Limitations of the study are due to the methodology of the test: only cytogenetic disorders in a single tissue were analyzed under conditions of double enteral administration of the studied sample. Conclusion. The sample of the food dye Tartrazine (E 102) in the dose range of 250-2000 mg/kg did not show cytogenetic activity in the in vivo micronucleus test on mouse bone marrow cells after a double enteral administration.

Single-cell RNA-seq of primary bone marrow neutrophils from female and male adult mice

Widespread sex-dimorphism is observed in the mammalian immune system. Consistently, studies have reported sex differences in the transcriptome of immune cells at the bulk level, including neutrophils. Neutrophils are the most abundant cell type in human blood, and they are key components of the innate immune system as they form a first line of defense against pathogens. Neutrophils are produced in the bone marrow, and differentiation and maturation produce distinct neutrophil subpopulations. Thus, single-cell resolution studies are crucial to decipher the biological significance of neutrophil heterogeneity. However, since neutrophils are very RNA-poor, single-cell profiling of these cells has been technically challenging. Here, we generated a single-cell RNA-seq dataset of primary neutrophils from adult female and male mouse bone marrow. After stringent quality control, we found that previously characterized neutrophil subpopulations can be detected in both sexes. Additionally, we confirmed that canonical sex-linked markers are differentially expressed between female and male cells across neutrophil subpopulations. This dataset provides a groundwork for comparative studies on the lifelong transcriptional sexual dimorphism of neutrophils.

Effects of Bone Marrow Autotransplantation on the Bone Marrow Neurotransmitter Structures.

Morphological structures involved in local regulation of tissue processes and the neurotransmitters they produce are assumed to play an important role in the pathogenesis of oncological diseases. We studied the dynamics of neurotransmitter cell counts and distribution of catecholamines and serotonin in them and analyzed proliferative activity of hemopoietic cells in the bone marrow of mice after bone marrow autotransplantation. The bone marrow of male mice was studied by fluorescent and immunohistochemical methods after injection of the bone marrow from the same mouse into the caudal vein. The count of neurotransmitter cells and the cell proliferation index increased after bone marrow autotransplantation.

Acute and chronic waves of myeloid cell maturation and myelopoiesis with high fat diet

Abstract Diet-induced obesity is associated with an infiltration of monocytes into the adipose tissue in male mouse models. These monocytes differentiate into inflammatory macrophages that drive insulin resistance. This increase in circulating monocytes originates in the bone marrow (BM) where hematopoietic stem cells (HSC) and myeloid-restricted progenitors (GMPs and PreGMs) expand. HSCs isolated from BM of male mice on high-fat diet (HFD) for 16 weeks show myeloid lineage skewing that lasts through successive bone marrow transplants (BMT). This suggests that HFD induces changes to the HSC niche - a multicellular structure that produces regulatory signals that are critical for maintaining the HSC pool. While BM adipocytes expand with HFD exposure, the niche cells promoting myelopoiesis with HFD are unknown. We chose to investigate two cell populations that have been implicated in driving myelopoiesis in other conditions; the BM endothelial cell (BMEC), a critical HSC niche cell, and neutrophils. We hypothesize that HFD induces HSC expansion and reprogramming through neutrophil-BMEC communication. Using 60% HFD in C57Bl/6J mice we evaluated circulating and BM neutrophils and Ly6C monocytes; HSCs and progenitors, and BMECs during short- and long-term HFD exposure. We found that as early as 24 hours after mice went on HFD there is a significant spike in circulating mature neutrophils that return to normal levels by 72 hours only to expand again by 16 weeks HFD. Ly6Chi monocytes rose only after 1 month of HFD exposure. BMECs do not appear to be altered in number with HFD in short- or long-term assays. Further qualitative analysis of BMECs, MAT, and neutrophils are necessary to determine which cells are critical for regulating the HSC niche in HFD.

Minimal uranium accumulation in lymphoid tissues following an oral 60-day uranyl acetate exposure in male and female C57BL/6J mice.

High levels of uranium (U) exist in soil, water, and air in the Southwestern United States due, in part, to waste generated from more than 160,000 abandoned hard rock mines located in this region. As a result, many people living in this region are chronically exposed to U at levels that have been linked to detrimental health outcomes. In an effort to establish a relevant in vivo mouse model for future U immunotoxicity studies, we evaluated the tissue distribution of U in immune organs; blood, bone marrow, spleen, and thymus, as well as femur bones, kidneys, and liver, following a 60-d drinking water exposure to uranyl acetate (UA) in male and female C57BL/6J mice. Following the 60-d exposure, there was low overall tissue retention of U (<0.01%) at both the 5 and the 50 ppm (mg/L) oral concentrations. In both male and female mice, there was limited U accumulation in immune organs. U only accumulated at low concentrations in the blood and bone marrow of male mice (0.6 and 16.8 ng/g, respectively). Consistent with previous reports, the predominant sites of U accumulation were the femur bones (350.1 and 399.0 ng/g, respectively) and kidneys (134.0 and 361.3 ng/g, respectively) of male and female mice. Findings from this study provide critical insights into the distribution and retention of U in lymphoid tissues following chronic drinking water exposure to U. This information will serve as a foundation for immunotoxicological assessments of U, alone and in combination with other metals.

Genotoxic Effect of Methotrexate on Bone Marrow Chromosomes and DNA of Male Albino Mice (Mus Musculus)

Aim of the work-Methotrexate (MTX), a structural analogue of folic acid, is an antineoplastic and antirheumatic agent which is used in a variety of clinical schedules and combination therapy regimens in man. Material and methods- Sixty mice of nearly the same age were randomly categorized into four groups (one control and three treated groups with different doses of methotrexate). Mice of the treated groups 1, 2 and 3 were intraperitoneally injected with a single dose of methotrexate (2.5, 5 or 10 mg/kg b. wt. respectively) at the first day of the experiment. All the control and the treated animals were sacrificed after 24, 48 or 72 hour by cervical dislocation post treatment. Results-Methotrexate treatment induced structural and numerical chromosomal aberrations in male mice bone marrow cells which were significantly increased (P< 0.001) by dose and time. Structural aberrations were chromosomal gap, fragment, break, centromeric attenuation, deletion, centric fusion, ring formation, end to end association and beaded chromosomes. Numerical aberration was polyploidy. Also, methotrexate treatment decreased the mitotic index in bone marrow cells of all the treated mice in comparison with the control group by increasing dose and time of treatment. Comet assay results indicated that treatment with methotrexate significantly increased (P< 0.001) DNA damage in the blood leukocytes in dose and time dependent manner. Conclusion- It can be concluded that methotrexate induced genetic damage on the chromosomes and DNA content of male albino mice even after single treatment with low doses.

Toxicity studies of 5-amino-o-cresol administered dermally to F344/NTac rats and B6C3F1/N mice.

5-Amino-o-cresol is used as an oxidative dye coupler (secondary intermediate) or oxidative (permanent) in hair dye formulations. It was nominated for study by the National Cancer Institute because it is a widely used genotoxic hair dye component for which no cancer studies have been reported. Male and female F344/NTac rats and B6C3F1/N mice were administered 5-amino-o-cresol (greater than 99% pure) dermally for 3 months. Genetic toxicology studies were conducted in Salmonella typhimurium, peripheral blood erythrocytes of male and female mice, and bone marrow of male mice. (Abstract Abridged).

Chemosignals from isolated females have antimutagenic effect in dividing the cells of bone marrow from male mice of the CBA strain

A level of X-ray induced mitotic disturbances in the cells of the bone marrow of male mice was studied under the modifying influence ofchemosignals from isolated adult female mice of the CBA line. It has been shown that the frequency of chromosomal aberrations in irradiated (4 Gr) males after exposing them for 24 hours on bedding soiled with female chemosignals is lower than in irradiated males in cages with clean bedding. The mechanisms and importance of the antimutagenic effect of female house mouse chemosignals are discussed.

In Vivo Tracking and Comparison of the Therapeutic Effects of MSCs and HSCs for Liver Injury

BackgroundMesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) have been studied for damaged liver repair; however, the conclusions drawn regarding their homing capacity to the injured liver are conflicting. Besides, the relative utility and synergistic effects of these two cell types on the injured liver remain unclear.Methodology/Principal FindingsMSCs, HSCs and the combination of both cells were obtained from the bone marrow of male mice expressing enhanced green fluorescent protein(EGFP)and injected into the female mice with or without liver fibrosis. The distribution of the stem cells, survival rates, liver function, hepatocyte regeneration, growth factors and cytokines of the recipient mice were analyzed. We found that the liver content of the EGFP-donor cells was significantly higher in the MSCs group than in the HSCs or MSCs+HSCs group. The survival rate for the MSCs group was significantly higher than that of the HSCs or MSCs+HSCs group; all surpassed the control group. After MSC-transplantation, the injured livers were maximally restored, with less collagen than the controls. The fibrotic areas had decreased to a lesser extent in the mice transplanted with HSCs or MSCs+HSCs. Compared with mice in the HSCs group, the mice that received MSCs had better improved liver function. MSCs exhibited more remarkable paracrine effects and immunomodulatory properties on hepatic stellate cells and native hepatocytes in the treatment of the liver pathology. Synergistic actions of MSCs and HSCs were most likely not observed because the stem cells in liver were detected mostly as single cells, and single MSCs are insufficient to provide a beneficial niche for HSCs.Conclusions/SignificanceMSCs exhibited a greater homing capability for the injured liver and modulated fibrosis and inflammation more effectively than did HSCs. Synergistic effects of MSCs and HSCs were not observed in liver injury.

Amelioration of Murine Schistosoma mansoni Induced Liver Fibrosis by Mesenchymal Stem Cells

Schistosomiasis is a common chronic helminthic infection of the liver that causes hepatic fibrosis and portal hypertension,contributing to the death of over half a million people a year. Infusion of autologous bone marrow cells into patients with hepatic cirrhosis has been reported to ameliorate symptoms of portal hypertension and improve liver function, either by conversion of the infused mesenchymal stem cells (MSCs) to hepatocytes or by modulating of the hepatic fibrosis. Here,we have investigated the antifibrotic effect of mesenchymal stem cells (MSCs) using S. mansoni-induced liver fibrosis in mice, which causes an intense, stable fibrosis. MSCs derived from bone marrow of male mice were then infused intravenously into female mice that had received intraperitoneal injection of S.mansoni cercariae. Mice were divided into 4 groups: Untreated control; MSCs infusion only; Schistosomiasis only; and Schistosomiasis plus MSCs infusion. Serum alanine aminotransferase (ALT) and liver histopathology were evaluated. Expression of the collagen gene (type I),transforming growth factor (TGF-β), matrix metalloproteinase (MMP2), tissue inhibitor of metalloproteinase (TIMP-1),stromal cell-derived factor-1(SDF-1) and its receptor (CXCR4) were analyzed. MSC infusion resulted in significant decrease in liver collagen and TGF-β gene expression in the Schistosomiasis mice. The ratio of MMP-2 to TIMP-1 expression increased. SDF-1 and CXCR4 mRNA expression also increased. There was overall improvement of liver histology and a statistically significant reduction of serum ALT level. MSCs infusion ameliorated S. mansoni-induced liver fibrosis, probably by modulating the relative expression of MMP and TIMP. The findings support the hypothesis that MSCs participate in liver regeneration and functional improvement by reducing liver fibrosis.

Differential anti-inflammatory and anti-fibrotic activity of transplanted mesenchymal vs. hematopoietic stem cells in carbon tetrachloride-induced liver injury in mice

Bone marrow stem cells nullify acquired and non-acquired diseases of liver through multiple strategies including antiinflammation. However, little is known about the in vivo mechanism of immunomodulation by stem cells in mediating liver cirrhosis. Mesenchymal stem cells (MSC) or hematopoietic stem cells (HSC) isolated from bone marrow of male mice were transplanted into female mice with acute liver inflammation. Serum levels of liver proteins and aminotransferase as well as hepatic antioxidant enzymes were estimated. Immunostaining for the expression of tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), alpha smooth muscle actin (α-SMA) and type I collagen proteins was carried out and the expression of these mRNAs was also studied. After post-transplantation, the levels of serum albumin and aminotransferases became normal and the levels of antioxidants were significantly high in the MSC treated mice compared to HSC and control mice. Necrotic cells and invasion of neutrophils were not observed in histological sections of liver of MSC treated mice. Immunostaining showed that IL-6 and TNF-α were not expressed in the MSC treated mice when compared to the control and HSC treated mice. α-SMA representing activated myofibroblasts and type I collagen were not expressed in MSC treated group. These inflammatory and fibrogenic results were further confirmed by reverse transcription-polymerase chain reaction (RT-PCR). The acute inflammation ended with the formation of fibrosis in the HSC and control groups by the uncontrolled immunoreactions. Protection mechanism of MSC therapy against injury and fibrosis in the liver occurs by the suppression of inflammation. Our findings suggest that bone marrow MSC are capable of alleviating the immunoreactions leading to the fibrosis in the liver.

Gene expression of SDF-1 in the derma wound healing in three kinds of different immune state mice

Objective To explore gene transcription of stromal cell derived factor-1 (SDF-1) in the derma wound healing in three kinds of different immune state mice. Methods BMSCs were isolated from BALB/C male mice bone marrow and further purified according to the different adhenrence of different kinds of cells to the wall of culture flask. The third generation of BMSCs was used for inducing osteogenesis and adipogenesis. Then, BALB/C,Nude and SCID female mice were transplanted with other third generation of male BMSCs via the tail vein after 3.5 Gy irradiation. The back skin wound was constructed in the bone marrow-chimeric mice in 4 hours after irradiation. A piece of wound tissue was harvested in post-operation 1,3,5,7 and 14 days in order to detect mRNA (messenger RNA) expressions of SDF-1 at different time points by Reverse t ranscription-polymerase chain reaction (RT-PCR) analysis. Results Osteogenesis and adipogenesis could be induced from BALB/C male mice BMSCs in vitro. During wound healing in BALB/C mice,SDF-1 expression began increasing on day 1 after injury,reached the peak level on day 5 (P<0.01) ,and then decreaseded gradually,when wound healing on day 14,SDF-1 level was near to its control group; while SDF-1 expression up-regulated on day 1 after injury in Nude and SCID mice,reached the peak level on day 7(P<0. 01 ) ,then decreaseded gradually, contrasting to BALB/C mice,peaking of SDF-1 was delayed,and their wounds could be healed on day 14,SDF-1 level exceeded their control group. Conclusion SDF-1 can enhance BMSCs mobilization and recruitment in the wound, participate in the period of inflammation and proliferation,and it plays a pivotal role during wound healing; immune function of organism can influence BMSCs recruitment, thus influence wound healing. Key words: Stroma-cell derived factor-1; Tauma and injury; Wound healing; Gene expression

Absence of DNA Damage in Comet Assay and Liver Initiation Activity of Cinnamaldehyde

To clarify the in vivo genotoxicity of cinnamaldehyde (CNMA) that is present in cigarette smoke, 8 organs and bone marrow of male mice given a single oral administration of 250 or 125 mg/kg body weight of CNMA were evaluated by an alkaline single-cell gel electrophoresis (comet) assay. The liver initiation activity of CNMA was also investigated by a short-term liver initiation assay. PH (Partially hepatectomized) male rats were given 1250 mg/kg body weight of CNMA by a single oral administration 12 hours after partial hepatectomy. In the comet assay, no significant differences of nuclear migration were observed in all tissues examined in mice treated with CNMA at 3 and 24 hours after administration. In the initiation assay, there were no significant increases in areas and numbers of glutathione S-transferase placental form (GST-P) positive foci in the livers of rats treated with CNMA. Based on these results, the possibility that CNMA has in vivo genotoxicity or liver initiation activity appears to be extremely low.

Protective Effect Of Ascorbic Acid On Cisplatin Genotoxicity In Male Mice Bone Marrow Cells

Cisplatin (cis-diamminedichloroplatinum II) is an effective antitumor agent with a wide spectrum of activity against varies solid tumors, but it has serious side effects on nontumour cells. Cisplatin produces intra- and interstrand DNA cross-linking effects and chromosomal aberrations in mammalian cells. Vitamin C (ascorbic acid) is an antioxidant that can scavenge free radicals and protect cellular macromolecules, including DNA, from oxidative damage induced by different agents. Pretreatment administration of ascorbic acid on cisplatin induced chromosome aberrations has been determined in bone marrow cells of Swiss albino mice. Results showed that cisplatin (7.5 & 10mg/kg bw)IP injection to male mice induced significant increase in the frequencies of chromosomal aberrations. The results of pre-treatment with ascorbic acid (66mg/kg bw) showed a significant decrease in the number of chromosomal aberrations induced with cisplatin tested doses. Ascorbic acid did not exhibit any clastogenic effect in male mice bone marrow cells. We concluded that ascorbic acid has a protective role against the genotoxicity induced by antitumor drug cisplatin.

Protective Effect Of Ascorbic Acid On Cisplatin Genotoxicity In Male Mice Bone Marrow Cells

Cisplatin (cis-diamminedichloroplatinum II) is an effective antitumor agent with a wide spectrum of activity against varies solid tumors, but it has serious side effects on nontumour cells. Cisplatin produces intra- and interstrand DNA cross-linking effects and chromosomal aberrations in mammalian cells. Vitamin C (ascorbic acid) is an antioxidant that can scavenge free radicals and protect cellular macromolecules, including DNA, from oxidative damage induced by different agents. Pretreatment administration of ascorbic acid on cisplatin induced chromosome aberrations has been determined in bone marrow cells of Swiss albino mice. Results showed that cisplatin (7.5 & 10mg/kg bw)IP injection to male mice induced significant increase in the frequencies of chromosomal aberrations. The results of pre-treatment with ascorbic acid (66mg/kg bw) showed a significant decrease in the number of chromosomal aberrations induced with cisplatin tested doses. Ascorbic acid did not exhibit any clastogenic effect in male mice bone marrow cells. We concluded that ascorbic acid has a protective role against the genotoxicity induced by antitumor drug cisplatin.

Micronucleus formation induced by the combination of low doses of X-rays and antineoplastic drugs in bone marrow of male mice.

People are widely exposed during their lifetime to many biological, chemical, and physical agents in the environment and at work. In this paper the effects of combined exposures of nonmutagenic doses of X-rays and anticancer agents (cyclophosphamide, mitomycin C, and vinblastine) have been investigated on the induction of micronuclei in the bone marrow of laboratory mice. The combination of X-rays and anticancer drugs enhanced the frequency of micronuclei in some cases. The strongest effects were found after the combination of X-rays and cyclophosphamide at 24 h and 72 h. The combined treatment of X-rays and mitomycin C enhanced the mutagenic effect at 72 h. The combination of X-rays + vinblastine slightly potentiated the mutagenic effect at 24 h and 48 h.

Mutagenicity to the mouse bone marrow by the mouse germ cell mutagen N-propyl-N-nitrosourea

N-Propyl-N-nitrosourea (PNU) is shown to be active in male mouse bone marrow micronucleus assays when dosed at either 100 or 200 mg/kg in saline. Activity was observed following either intraperitoneal (i.p.) injection or oral gavage. This observation is consistent with the demonstration by Murota and Shibuya of the specific-locus mutagenicity caused by PNU in male mouse spermatogonia when dosed at 200 mg/kg by i.p. injection. These data strengthen further the observation that rodent germ cell mutagens are also mutagenic to rodent somatic cells.

Examination of the mononuclear phagocyte system in lupus-prone male BXSB mice.

During the course of lupus-like autoimmune disease male BXSB mice develop an increasing monocytosis in the peripheral blood. As we demonstrated previously, this monocytosis is parallelled by an expansion of a strain-specific high number of macrophage precursor cells (CFU-M) in the bone marrow of male mice. To test the hypothesis that the expanded mononuclear phagocyte system (MPS) may promote autoimmune disease in these mice the organ-associated macrophage system was examined. Our latest data show that an unusual expansion of CFU-M also appears in spleen and liver of male mice 2 weeks after birth. In addition to a morphological alteration of the organs during the course of the disease there is a change in number and distribution of organ-resident macrophages. Considering these results the possible contribution of the expanded MPS in promoting autoimmune disease is discussed.

Treatment of mice with a novel antineoplastic agent taxol before irradiation increases the frequency of micronuclei in the bone marrow

The frequency of micronucleated polychromatic erythrocytes (MPCE) and the normochromatic erythrocytes (MNCE) and polychromatic and normochromatic erythrocyte ratio ( P N ratio) was studied at 12, 24 and 36 h postirradiation in the bone marrow of male mice treated or not with taxol before exposure to 0–4 Gy of 60Co gamma radiation. The frequency of MPCE increased with the increase in radiation dose in a dose-related manner in the irradiated control group. A peak frequency of MPCE was observed at 24 h postirradiation in irradiated control group. The pattern of increase in MNCE was similar to that of MPCE except that a highest number of MNCE was scored at 36 h postirradiation. Taxol administration to animals before irradiation resulted in a significant elevation in the frequency of MPCE and MNCE at all the postirradiation time periods studied. This increase was dose related as observed in the irradiated control group. Irradiation resulted in a dose-dependent decline in the P N ratio at all the postirradiation time periods studied. The P N ratio was significantly lower in the taxol + irradiated group compared to the irradiated control group at all postirradiation time periods. A maximum decline in P N ratio was observed at 36 h postirradiation for both irradiated control and taxol + irradiated groups. The dose response for MPCE, MNCE and P N ratio was linear quadratic for both the irradiated and taxol + irradiated groups.

The effect of the male contraceptive agent gossypol acetic acid on mouse bone marrow cells in vivo: Micronuclei and mitotic index

In this work the genotoxic effect of gossypol acetic acid (gossypol) was evaluated by determining the frequency of micronuclei and mitotic index in male mouse bone marrow cells in vivo. Bone marrow cells were collected at 24th hour after the single intraperitoneal (20, 40, and 80 μg/g) administration of gossypol. Polychromatic erythrocytes (PCEs) in the bone marrow were then evaluated with respect to micronuclei frequency. The dose-dependent increase in the micronuclei frequency was observed. However, when compared with the control group, the increase was not found to be significiant (P > 0.05). Also the mitotic index values were not found to be different from those control values(P > 0.05). The results suggest that gossypol is not a clastogenic and mutagenic agent in mouse bone marrow cells in vivo.