scholarly journals Osteoporosis Affects Functional Activity and Gene Expression of Osteoblastic Cells Derived from Rat Alveolar Bone

2020 ◽  
Vol 31 (6) ◽  
pp. 617-622
Author(s):  
Paula Katherine Vargas-Sanchez ◽  
Roger Rodrigo Fernandes ◽  
Flávia Aparecida Chaves Furlaneto ◽  
Luiz Gustavo de Sousa ◽  
Selma Siéssere ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Alveolar Bone ◽  
Bone Cells ◽  
Female Rats ◽  
Ovariectomized Rats ◽  
Osteoblastic Cells ◽  
Expression Of Genes ◽  
In Situ Detection

Abstract Recent studies suggest that osteoporosis, in addition to the damage caused in long bones, may cause deterioration in the jaws, especially in alveolar bone sites, with effects in the progress of periodontal disease as well as in bone healing. The aim of this study was to evaluate the effect of osteoporosis in the metabolism of rat alveolar bone osteoblasts. There were used 10 female rats divided in two experimental groups (Sham and OVX), which were ovariectomized and after 8 weeks euthanized to collect mandibular bone samples in order to isolate osteoblastic cells. The cells were cultured in 24-well plates to perform the in vitro experiments. After 7, 10 and 14 days, there were evaluated cell proliferation by MTT assay, in situ detection of alkaline phosphatase (ALP) as well as mineralized nodules and expression of genes associated to bone remodeling. Results showed that at 7, 10 and 14 days cell proliferation was lower for OVX group. In situ detection of ALP was higher at 7 days and lower at 10 and 14 days in OVX group. At 17 and 21 days, OVX group had a significative decrease of mineralization nodules. There was a downregulation in the expression of Alp, Bglap and Runx2 genes and an upregulation of Opg in OVX group, whereas Opn and Rankl modulation was similar between the evaluated groups. Our results suggest that osteoporosis has a deleterious effect on alveolar bone cells from ovariectomized rats, which might affect the treatment of diseases associated to the jaw bones.

scholarly journals In Situ Detection of Mycobacterium tuberculosis Transcripts in Human Lung Granulomas Reveals Differential Gene Expression in Necrotic Lesions

2002 ◽  
Vol 70 (11) ◽  
pp. 6330-6338 ◽  
Author(s):  
Gael Fenhalls ◽  
Liesel Stevens ◽  
Lorraine Moses ◽  
Juanita Bezuidenhout ◽  
Joanna C. Betts ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mycobacterium Tuberculosis ◽  
Transition Zone ◽  
Expression Of Genes ◽  
Metabolic States ◽  
Necrotic Region ◽  
Rna In Situ Hybridization ◽  
In Situ Detection

ABSTRACT We have used RNA-RNA in situ hybridization to detect the expression of several Mycobacterium tuberculosis genes in tuberculous granulomas in lung tissue sections from tuberculosis patients. The M. tuberculosis genes chosen fall into two classes. Four genes (icl, narX, and Rv2557 and Rv2558) have been implicated in the persistence of the bacterium in the host, and two genes (iniB and kasA) are upregulated in response to isoniazid exposure. Both necrotic and nonnecrotic granulomas were identified in all of the patients. Necrotic granulomas were divided into three zones: an outer lymphocyte cuff containing lymphocytes and macrophages, a transition zone consisting of necrotic material interspersed with macrophages, and a central acellular necrotic region. Transcripts of all of the genes studied were found in nonnecrotic granulomas and in the lymphocyte cuff of necrotic granulomas. Mycobacterial gene expression was associated with CD68-positive myeloid cells. Rv2557 and/or its homologue Rv2558, kasA, and iniB were expressed within the transition zone of necrotic granulomas, whereas icl and narX transcripts were absent from this area. There was no evidence of transcription of any of the genes examined in the central necrotic region, although mycobacterial DNA was present. The differential expression of genes within granulomas demonstrates that M. tuberculosis exists in a variety of metabolic states and may be indicative of the response to different microenvironments. These observations confirm that genes identified in models of persistence or in response to drug treatment in vitro are expressed in the human host.

Prolactin decreases expression of Runx2, osteoprotegerin, and RANKL in primary osteoblasts derived from tibiae of adult female rats

2008 ◽  
Vol 86 (5) ◽  
pp. 240-248 ◽  
Author(s):  
Narattaphol Charoenphandhu ◽  
Jarinthorn Teerapornpuntakit ◽  
Methajit Methawasin ◽  
Kannikar Wongdee ◽  
Kanogwun Thongchote ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Bone Cells ◽  
Nuclear Factor Κb ◽  
Clinical Findings ◽  
Female Rats ◽  
Adult Rats ◽  
Expression Of Genes ◽  
Primary Osteoblasts ◽  
Underlying Mechanisms

Hyperprolactinemia caused by physiological or pathological conditions, such as those occurring during lactation and prolactinoma, respectively, results in progressive osteopenia. The underlying mechanisms, however, are controversial. Prolactin (PRL) may directly attenuate the functions of osteoblasts, since these bone cells express PRL receptors. The present study therefore aimed to investigate the effects of PRL on the expression of genes related to the osteoblast functions by using quantitative real-time PCR technique. Herein, we used primary osteoblasts that were derived from the tibiae of adult rats and displayed characteristics of differentiated osteoblasts, including in vitro mineralization. Osteoblasts exposed for 48 h to 1000 ng/mL PRL, but not to 10 or 100 ng/mL PRL, showed decreases in the mRNA expression of Runx2, osteoprotegerin (OPG), and receptor activator of nuclear factor κB ligand (RANKL) by 60.49%, 72.74%, and 87.51%, respectively. Nevertheless, PRL did not change the RANKL/OPG ratio, since expression of OPG and RANKL were proportionally decreased. These concentrations of PRL had no effect on the mRNA expression of osteocalcin and osteopontin, nor on mineralization. High pathologic concentrations of PRL (1000 ng/mL) may downregulate expression of genes that are essential for osteoblast differentiation and functions. The present results explained the clinical findings of hyperprolactinemia-induced bone loss.

Bone Invasive Properties of Oral Squamous Cell Carcinoma and its Interactions with Alveolar Bone Cells: An In Vitro Study

2019 ◽  
Vol 19 (8) ◽  
pp. 631-640 ◽  
Author(s):  
Omel Baneen Qallandar ◽  
Faeza Ebrahimi ◽  
Farhadul Islam ◽  
Riajul Wahab ◽  
Bin Qiao ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Oral Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cancer Cells ◽  
Alveolar Bone ◽  
Cultured Cells ◽  
Bone Cells ◽  
Bone Invasion ◽  
Alveolar Bone Cells

Background: Co-culture of cancer cells with alveolar bone cells could modulate bone invasion and destructions. However, the mechanisms of interaction between oral squamous cell carcinoma (OSCC) and bone cells remain unclear. Objective: The aim of this study is to analyse the direct and indirect effects of OSCC cells in the stimulation of osteolytic activity and bone invasion. Methods: Direct co-culture was achieved by culturing OSCC (TCA8113) with a primary alveolar bone cell line. In the indirect co-culture, the supernatant of TCA8113 cells was collected to culture the alveolar bone cells. To assess the bone invasion properties, in vitro assays were performed. Results: The proliferation of co-cultured cancer cells was significantly (p<0.05) higher in comparison to the monolayer control cells. However, the proliferation rates were not significantly different between direct and indirect co-cultured cells with indirect co-cultured cells proliferated slightly more than the direct co-cultured cells. Invasion and migration capacities of co-cultured OSCC and alveolar bone cells enhanced significantly (p<0.05) when compared to that of control monolayer counterparts. Most importantly, we noted that OSCC cells directly co-cultured with alveolar bone cells stimulated pronounced bone collagen destruction. In addition, stem cells and epithelialmesenchymal transition markers have shown significant changes in their expression in co-cultured cells. Conclusion: In conclusion, the findings of this study highlight the importance of the interaction of alveolar bone cells and OSCC cells in co-culture setting in the pathogenesis of bone invasion. This may help in the development of potential future biotherapies for bone invasion in OSCC.

scholarly journals Peripheral administration of GH induces cell proliferation in the brain of adult hypophysectomized rats

2009 ◽  
Vol 201 (1) ◽  
pp. 141-150 ◽  
Author(s):  
N David Åberg ◽  
Inger Johansson ◽  
Maria A I Åberg ◽  
Johan Lind ◽  
Ulf E Johansson ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Corpus Callosum ◽  
Progenitor Cells ◽  
Parietal Cortex ◽  
Cellular Proliferation ◽  
Piriform Cortex ◽  
Adult Brain ◽  
Female Rats ◽  
Igf I

IGF-I treatment has been shown to enhance cell genesis in the brains of adult GH- and IGF-I-deficient rodents; however, the influence of GH therapy remains poorly understood. The present study investigated the effects of peripheral recombinant bovine GH (bGH) on cellular proliferation and survival in the neurogenic regions (subventricular zone (SVZ), and dentate gyrus of the hippocampus), as well as the corpus callosum, striatum, parietal cortex, and piriform cortex. Hypopituitarism was induced in female rats by hypophysectomy, and the rats were supplemented with thyroxine and cortisone acetate. Subsequently, the rats received daily s.c. injections of bGH for either 6 or 28 days respectively. Following 5 days of peripheral bGH administration, the number of bromodeoxyuridine (BrdU)-positive cells was increased in the hippocampus, striatum, parietal cortex, and piriform cortex after 6 and 28 days. In the SVZ, however, BrdU-positive cells increased only after 28 days of bGH treatment. No significant change was observed in the corpus callosum. In the hippocampus, after 28 days of bGH treatment, the number of BrdU/NeuN-positive cells was increased proportionally to increase the number of BrdU-positive cells. 3H-thymidine incorporation in vitro revealed that 24 h of bGH exposure was sufficient to increase cell proliferation in adult hippocampal progenitor cells. This study shows for the first time that 1) peripheral bGH treatment increased the number of newborn cells in the adult brain and 2) bGH exerted a direct proliferative effect on neuronal progenitor cells in vitro.

scholarly journals Trophectoderm-Specific Knockdown of LIN28 Decreases Expression of Genes Necessary for Cell Proliferation and Reduces Elongation of Sheep Conceptus

2020 ◽  
Vol 21 (7) ◽  
pp. 2549 ◽  
Author(s):  
Asghar Ali ◽  
Mark Stenglein ◽  
Thomas Spencer ◽  
Gerrit Bouma ◽  
Russell Anthony ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Critical Period ◽  
In Vitro Experiments ◽  
Placental Development ◽  
Expression Of Genes ◽  
Trophectoderm Cells ◽  
Successful Establishment

LIN28 inhibits let-7 miRNA maturation which prevents cell differentiation and promotes proliferation. We hypothesized that the LIN28-let-7 axis regulates proliferation-associated genes in sheep trophectoderm in vivo. Day 9-hatched sheep blastocysts were incubated with lentiviral particles to deliver shRNA targeting LIN28 specifically to trophectoderm cells. At day 16, conceptus elongation was significantly reduced in LIN28A and LIN28B knockdowns. Let-7 miRNAs were significantly increased and IGF2BP1-3, HMGA1, ARID3B, and c-MYC were decreased in trophectoderm from knockdown conceptuses. Ovine trophoblast (OTR) cells derived from day 16 trophectoderm are a useful tool for in vitro experiments. Surprisingly, LIN28 was significantly reduced and let-7 miRNAs increased after only a few passages of OTR cells, suggesting these passaged cells represent a more differentiated phenotype. To create an OTR cell line more similar to day 16 trophectoderm we overexpressed LIN28A and LIN28B, which significantly decreased let-7 miRNAs and increased IGF2BP1-3, HMGA1, ARID3B, and c-MYC compared to control. This is the first study showing the role of the LIN28-let-7 axis in trophoblast proliferation and conceptus elongation in vivo. These results suggest that reduced LIN28 during early placental development can lead to reduced trophoblast proliferation and sheep conceptus elongation at a critical period for successful establishment of pregnancy.

Vasopressin responses and receptors in the mesenteric vasculature of estrogen-treated rats

1986 ◽  
Vol 251 (5) ◽  
pp. H885-H889 ◽  
Author(s):  
J. St-Louis ◽  
A. Parent ◽  
R. Lariviere ◽  
E. L. Schiffrin
Keyword(s):  
Binding Sites ◽  
Pressor Response ◽  
Female Rats ◽  
Maximum Response ◽  
Male Rats ◽  
Ovariectomized Rats ◽  
Binding Characteristics ◽  
Vascular Bed ◽  
Mesenteric Vascular Bed

The effect of treatment with estrogens on the biological activity of arginine8 vasopressin (AVP) in the in vitro perfused mesenteric vascular bed and on the binding characteristics of [3H]AVP on membranes prepared from the same vascular bed was studied. Female rats treated with estradiol (400 micrograms/24 h sc), compared with ovariectomized rats, had an increase in the maximum response to AVP (from 128 +/- 3 to 153 +/- 3 mmHg) in the perfused preparation and an increase in the density of AVP binding sites (from 402 to 732 fmol/mg protein) in the membrane preparation. In male rats, the injection of estradiol increased the maximum response to AVP (from 109 +/- 4 to 137 +/- 3 mmHg) and the density of AVP binding sites (from 289 to 519 fmol/mg protein). The effective concentration producing 50% of maximum response of AVP in the perfused preparation was higher in male than in female rats, while the Kd in the binding experiments was similar in the four experimental groups. Our results show that estrogens upregulate the number of AVP binding sites, leading to an increase in the pressor response to AVP in the rat mesenteric vascular bed.

Effect of Substrate Surface Modification on Biomineralization of Osteoblasts

2006 ◽  
Vol 950 ◽  
Author(s):  
Yizhi Meng ◽  
Xiaolan Ba ◽  
Seo-Young Kwak ◽  
Elaine DiMasi ◽  
Meghan Ruppel ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Bone Cells ◽  
Bone Mineralization ◽  
Substrate Surface ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Tissue Scaffold ◽  
Actin Fiber

ABSTRACTUnderstanding how biomineralization occurs in the extracellular matrix (ECM) of bone cells is crucial to the development of a successfully engineered bone tissue scaffold, and to date there has not been a well-established method for the quantitative examination of bone mineralization in situ. We investigated the mechanical properties of MC3T3-E1 osteoblast-like cells and the crystalline properties of their biomineralized ECM in vitro using shear modulation force microscopy (SMFM), confocal laser scanning microscopy (CLSM), synchrotron X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR). The elastic modulus of the mineralizing cells increased at time points corresponding to mineral production, whereas that of the non-mineralizing cells did not vary significantly over time. CLSM showed a restructuring of the F-actin fiber network of mineralizing cells with time, which indicates remodeling activities in the cytoskeleton and was not seen in the non-mineralizing cells. Both XRD and FTIR showed that the mineralizing subclone produced hydroxyapatite in situ and that the non-mineralizing subclone was in fact weakly biomineralizing.

IN VITRO AND IN SITU DETECTION OF ERWINIA AMYLOVORA WITH MONOCLONAL ANTIBODIES

1987 ◽  
pp. 77-80
Author(s):  
C.P. Lin ◽  
T.A. Chen ◽  
J.M. Wells ◽  
T. van der Zwet
Keyword(s):  
Monoclonal Antibodies ◽  
Erwinia Amylovora ◽  
In Situ Detection
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