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In Vitro Cellular & Developmental... Mar 2011Dimethyl sulfoxide (DMSO) is routinely used in the laboratory as a solvent and vehicle for organic molecules. Although it has been used in previous studies involving...
Dimethyl sulfoxide (DMSO) is routinely used in the laboratory as a solvent and vehicle for organic molecules. Although it has been used in previous studies involving myeloid cells and macrophages, we are unaware of data demonstrating the effects of DMSO alone on osteoclast development. Recently, we were using DMSO as a vehicle and included a non-vehicle control. Surprisingly, we observed a marked change in osteoclast development, and therefore designed this study to examine the effects of DMSO on osteoclast development. Osteoclasts were generated from two sources: bone marrow macrophages and an osteoclast progenitor cell line. Cells were cultured with DMSO for various durations and at differing concentrations and mature, multinucleated (>3 nuclei) TRAP(+) cells were assessed in terms of cell number, cell surface area, and number of nuclei/cell. Osteoclast surface area increased in 5 μM DMSO to a mean of 156,422 pixels from a mean of 38,510 pixels in control culture, and subsequently decreased in 10 μM DMSO to a mean of 18,994 pixels. With serial addition of DMSO over 5 d, a significant increase in mean surface area, and number of nuclei/cell was also observed, while the opposite was true when DMSO was serially removed from culture. These findings show that DMSO exerts a marked effect on osteoclast differentiation. Since many investigators use DMSO to solubilize compounds for treatment of osteoclasts, caution is warranted as altering DMSO concentrations may have a profound effect on the final data, especially if osteoclast differentiation is being assessed.
Topics: Animals; Cell Count; Dimethyl Sulfoxide; Mice; Osteoclasts; Surface Properties; Time Factors
PubMed: 21359822
DOI: 10.1007/s11626-011-9385-8 -
Connective Tissue Research 1989When implanted subcutaneously in rats, devitalized bone particles (BP) elicit the differentiation of osteoclastic cells. Those cells can be distinguished from foreign... (Review)
Review
When implanted subcutaneously in rats, devitalized bone particles (BP) elicit the differentiation of osteoclastic cells. Those cells can be distinguished from foreign body giant cells that form in response to particulate plastics. Osteoclast features include resorption of the bone substrate, ruffled borders, calcitonin receptors, tartrate-resistant acid phosphatase activity, and modulation by bone active agents. To determine whether expression of these features depends on specific components of the matrix, we characterized the multinucleated cells that developed in response to osteocalcin-deficient BPs, particulate microcrystalline hydroxyapatite (HA), and HA containing 0.1% osteocalcin, collagen, or bovine serum albumin. Only those particles that contained mineral and osteocalcin were associated with osteoclastic cells. These studies support the hypothesis that osteocalcin may function as a matrix signal in the differentiation of osteoclasts.
Topics: Animals; Bone Matrix; Cell Differentiation; Hydroxyapatites; Osteocalcin; Osteoclasts; Rats
PubMed: 2692953
DOI: 10.3109/03008208909023880 -
Zhongguo Gu Shang = China Journal of... Apr 2013Osteoclasts and osteoblasts are not exist alone,while communicating with each other through direct contact, diffusible paracrine factors and cell-bone matrix... (Review)
Review
Osteoclasts and osteoblasts are not exist alone,while communicating with each other through direct contact, diffusible paracrine factors and cell-bone matrix interaction. Co-culture system of osteoblast with osteoclast,including direct co-culture and indirect co-culture. It should be according to the ratio of osteoclasts and osteoblasts under the pathology, choosing the same species. Compared with lonely culture of osteoblasts or osteoclasts,co-culture system is much closer to the microenvironment in vivo. It benefits to explain the interactions between osteoblasts and osteoclasts, exploring molecular communication in bone diseases. It was mainly used to investigate the pharmacological mechanism of herbal and western medicine in bone remodeling. Some osteoporosis drugs (such as epimedium,sanchi, fructus psoraleae, ranelate strontium) not only promoted osteoblastic bone formation, but also inhibited osteoclastic bone resorption in the system,so as to balance bone homeostasis. At the same time,it has been used to study medical physics and assess biomedical materials in recent years. Considerably,the co-cultrue system will be used to study the subchondral bone remodeling and its pharmacological mechanism of herbal and western medicine in osteoarthritis.
Topics: Animals; Bone Remodeling; Cell Communication; Coculture Techniques; Humans; Osteoblasts; Osteoclasts
PubMed: 23844502
DOI: No ID Found -
Biochemical and Biophysical Research... Jan 2019Osteoporosis is a metabolic bone lesion in which the bone mass is reduced per unit volume due to increased bone resorption. Its main characteristics are bone pain and...
Osteoporosis is a metabolic bone lesion in which the bone mass is reduced per unit volume due to increased bone resorption. Its main characteristics are bone pain and increasing danger of fragility fracture. Excessive osteoclast activation is known to be responsible for extensive bone resorption. Thus, inhibition of osteoclastic bone resorption and regulation of the bone microenvironment are vital treatment strategies for osteoporosis. For the first time, we investigated the effect of proanthocyanidins (PACs) extracted from grape seed, which significantly inhibited osteoclast formation and differentiation from bone marrow macrophages (BMMs) and the RAW264.7 cell line and efficiently attenuated osteoclastic bone resorption without toxicity. These findings were confirmed by changes in the NF-κB and JNK/mitogen-activated protein kinase (MAPK) signaling pathways, which are major and classical signaling pathways involved in RANKL-mediated osteoclastogenesis in vitro. The PACs inhibited osteoclast formation and differentiation by inhibiting the NF-κB and JNK signaling pathways and might be useful for the treatment of osteoporosis.
Topics: Animals; Bone Resorption; Grape Seed Extract; MAP Kinase Kinase 4; Mice; Mice, Inbred C57BL; NF-kappa B; Osteoclasts; Osteogenesis; Osteoporosis; Proanthocyanidins; RAW 264.7 Cells; Signal Transduction
PubMed: 30583865
DOI: 10.1016/j.bbrc.2018.12.125 -
European Journal of Histochemistry : EJH Mar 2022Histochemical detection of tartrate-resistant acid phosphatase (TRAP) activity is a fundamental technique for visualizing osteoclastic bone resorption and assessing...
Histochemical detection of tartrate-resistant acid phosphatase (TRAP) activity is a fundamental technique for visualizing osteoclastic bone resorption and assessing osteoclast activity status in tissues. This approach has mostly employed colorimetric detection, which has limited quantification of activity in situ and co-labelling with other skeletal markers. Here we report simple colorimetric and fluorescent TRAP assays in zebrafish and medaka, two important model organisms for investigating the pathogenesis of bone disorders. We show fluorescent TRAP staining, utilising the ELF97 substrate, is a rapid, robust and stable system to visualise and quantify osteoclast activity in zebrafish, and is compatible with other fluorescence stains, transgenic lines and antibody approaches. Using this approach, we show that TRAP activity is predominantly found around the base of the zebrafish pharyngeal teeth, where osteoclast activity state appears to be heterogeneous.
Topics: Acid Phosphatase; Animals; Colorimetry; Isoenzymes; Osteoclasts; Tartrate-Resistant Acid Phosphatase; Zebrafish
PubMed: 35330553
DOI: 10.4081/ejh.2022.3369 -
Bioscience, Biotechnology, and... 2016Osteoporosis is a debilitating disease caused by decreased bone density. Compounds with anti-osteoclastic activity, such as bisphosphonates, may help in the prevention...
Osteoporosis is a debilitating disease caused by decreased bone density. Compounds with anti-osteoclastic activity, such as bisphosphonates, may help in the prevention and treatment of osteoporosis. Herein, we determined the inhibitory effects of ginger hexane extract (GHE) on receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. The results showed that GHE (1) suppressed osteoclast differentiation and the formation of actin rings; (2) inhibited the expression of Nfatc1, a master transcriptional factor for osteoclast differentiation, in a dose-dependent manner (10-20 μg/mL); and (3) inhibited other osteoclastogenesis-related genes, such as Oscar, Dc-stamp, Trap, and Mmp9. These findings suggest that GHE may be used to prevent and treat osteoporosis by inhibiting osteoclast differentiation.
Topics: Animals; Cell Differentiation; Cell Line; Zingiber officinale; Hexanes; Mice; Osteoclasts; Plant Extracts; RANK Ligand
PubMed: 26967638
DOI: 10.1080/09168451.2015.1127133 -
Biochemical and Biophysical Research... Mar 2020Low density lipoprotein receptor-related protein 1 (LRP1), a multifunctional cell surface protein, is expressed in bone marrow-derived macrophages. While LRP1 is thought...
Low density lipoprotein receptor-related protein 1 (LRP1), a multifunctional cell surface protein, is expressed in bone marrow-derived macrophages. While LRP1 is thought to be a suppressor of osteoclast differentiation at late stages, its function at early stages remains unclear. Here we demonstrate that Lrp1 stable knockdown by lentiviral short hairpin RNA in macrophage cell line RAW264 cells inhibited RANKL-induced osteoclast formation and osteoclastic master transcription factor Nfatc1 mRNA expression as assessed by quantitative RT-PCR. Furthermore, knockdown of the Lrp1 gene suppressed not only differentiation, but also proliferation, and inhibitory effects on osteoblastic ALP activity by osteoclast-derived humoral factors. Thus, we propose that LRP1 in macrophages is required for both differentiation into osteoclasts and osteoclast-osteoblast interactions.
Topics: Animals; Cell Communication; Cell Differentiation; Cell Proliferation; Gene Knockdown Techniques; Low Density Lipoprotein Receptor-Related Protein-1; Mice; Mice, Inbred C57BL; Osteoblasts; Osteoclasts; RAW 264.7 Cells
PubMed: 31964526
DOI: 10.1016/j.bbrc.2020.01.065 -
Clinical Calcium 2018Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. We have originally established...
Osteoclasts are bone-resorbing giant polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. We have originally established an advanced imaging system for visualizing the in vivo behavior of mature osteoclasts in living bone tissues with intravital multiphoton microscopy. By means of this system, we could grasp the real time-course of osteoclastic bone resorption, and identified two distinct functional states of differentiated osteoclasts, 'bone-resorptive' and 'non-resorptive'. Intravital imaging also revealed that various biologic drugs acted directly on mature osteoclasts during inflammatory bone destruction. In this review, we show the latest data of intravital imaging of osteoclast dynamics.
Topics: Animals; Bone and Bones; Cell Survival; Drug Evaluation, Preclinical; Humans; Inflammation; Osteoclasts
PubMed: 29371486
DOI: No ID Found -
Arthritis Research & Therapy 2009Osteoclast precursors arise from the CD14+ CD16- population in controls but details about cell surface marker expression and functional characteristics of these cells is...
Osteoclast precursors arise from the CD14+ CD16- population in controls but details about cell surface marker expression and functional characteristics of these cells is unknown, particularly in patients with inflammatory arthritis. In a recent issue of Arthritis, Research and Therapy, Lari and colleagues found that osteoclasts developed from a proliferative CD14+ CD16- subset in healthy controls. These cells took on the morphology of osteoclasts, expressed mRNA for osteoclast-related genes and excavated pits on bone wafers. These findings provide new insights into monocyte diversity and provide evidence that osteoclast precursors arise from a small proliferating monocyte population in controls. Additional studies are needed in patients with inflammatory arthritis.
Topics: Biomarkers; Blood Circulation; Cell Differentiation; Humans; Osteoclasts; Stem Cells
PubMed: 19591635
DOI: 10.1186/ar2707 -
Calcified Tissue International Nov 2020Iron overload is closely associated with osteoporosis, the potential cellular mechanism involved in decreased osteoblast differentiation and increased osteoclast...
Iron overload is closely associated with osteoporosis, the potential cellular mechanism involved in decreased osteoblast differentiation and increased osteoclast formation. However, the effect of iron overload on the biological behavior in osteocytes has not been reported. This study aims to investigate the changes of osteocytic activity, apoptosis, and its regulation on osteoclastogenesis in response to iron overload. MLO-Y4 osteocyte-like cells and primary osteocytes from mice were processed with ferric ammonium citrate (FAC) and deferoxamine (DFO), the conditioned medium (CM) was harvested and co-cultured with Raw264.7 cells and bone marrow-derived macrophages (BMDMs) to induce them to differentiate into osteoclasts. Osteocyte apoptosis, osteoclast differentiation, osteocytic gene expression and protein secretion of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) was examined. Excessive iron has a toxic effect on MLO-Y4 osteocyte-like cells. Increased cell apoptosis in MLO-Y4 cells and primary osteocytes was induced by iron overload. The osteoclastic formation, differentiation-related gene expression, and osteoclastic bone-resorption capability were significantly increased after treated with the CM from iron overload-exposed osteocytes. Excessive iron exposure significantly promoted the gene expression and protein secretion of the RANKL in MLO-Y4 cells. Addition of RANKL-blocking antibody completely abolished the increase of osteoclast formation and bone resorption capacity induced by the CM from osteocytes exposed to excessive iron. Moreover, the pan-caspase apoptosis inhibitor, QVD (quinolyl-valyl-O-methylaspartyl-[-2,6-difluorophenoxy]-methylketone) was used to inhibit osteocyte apoptosis. The results showed osteocyte apoptosis induced by iron overload was reduced by QVD and accompanied by the decrease of soluble RANKL (sRANKL) in supernatant. The increase of osteoclast formation and bone resorption capacity induced by the CM from osteocytes exposed to excessive iron was significantly decreased by QVD. These results indicated that iron overload-induced osteocyte apoptosis is required to regulate osteoclast differentiation by increasing osteocytic RANKL production. This study, for the first time, reveals the indirect effect of iron overload on osteoclast differentiation through regulating osteocytes.
Topics: Animals; Apoptosis; Cell Differentiation; Iron Overload; Mice; Osteoclasts; Osteocytes; Osteoprotegerin; RANK Ligand; RAW 264.7 Cells
PubMed: 32995951
DOI: 10.1007/s00223-020-00735-x