scholarly journals Acid Dentin Lysate Modulates Macrophage Polarization and Osteoclastogenesis In Vitro

Materials ◽  
2021 ◽  
Vol 14 (22) ◽  
pp. 6920
Author(s):  
Jila Nasirzade ◽  
Zahra Kargarpour ◽  
Layla Panahipour ◽  
Reinhard Gruber
Keyword(s):  
Bone Marrow ◽  
Macrophage Polarization ◽  
Hematopoietic Cells ◽  
Tartrate Resistant Acid Phosphatase ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Polycytidylic Acid ◽  
M1 Macrophage ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Dentin prepared from extracted teeth is used as autograft for alveolar bone augmentation. Graft consolidation involves the acid lysis of dentin thereby generating a characteristic paracrine environment. Acid lysate of dentin is mimicking this environment. Acid dentin lysate (ADL) potentially targets hematopoietic cells thereby affecting their differentiation towards macrophages and osteoclasts; however, the question remains if ADL controls macrophage polarization and osteoclastogenesis. Here, we show that ADL reduced lipopolysaccharide (LPS)-induced macrophage polarization of the pro-inflammatory (M1) phenotype, indicated by attenuated Interleukin 1 (IL1), Interleukine 6 (IL6)and cyclooxygenase 2 (COX2) expression. This decrease in M1 macrophages was confirmed by the reduced phosphorylation and nuclear translocation of p65 in the LPS-exposed RAW 264.7 macrophages. Similarly, when RAW 264.7 macrophages were incubated with other agonists of Toll-like receptor (TLR) signaling e.g., FSL1, Polyinosinic-polycytidylic acid High Molecular Weight (Poly (1:C) HMW), Pam3CSK4, and imiquimod, ADL reduced the IL6 expression. We further show herein that ADL decreased osteoclastogenesis indicated by the reduced formation of multinucleated cell expressing cathepsin K and tartrate-resistant acid phosphatase in murine bone marrow cultures. Overall, our results suggest that acid dentin lysate can affect the differentiation of hematopoietic cells to M1 macrophage polarization and a decrease in osteoclastogenesis in bone marrow cultures.

Clonal succession of hematopoietic cells in long-term bone marrow cultures

1985 ◽  
Vol 99 (3) ◽  
pp. 348-350
Author(s):  
O. A. Gurevich ◽  
N. I. Drize ◽  
G. A. Udalov ◽  
I. L. Chertkov
Keyword(s):  
Bone Marrow ◽  
Hematopoietic Cells ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

scholarly journals Inhibin Suppresses and Activin Stimulates Osteoblastogenesis and Osteoclastogenesis in Murine Bone Marrow Cultures

Endocrinology ◽  
2002 ◽  
Vol 143 (1) ◽  
pp. 74-83 ◽  
Author(s):  
D. Gaddy-Kurten ◽  
J. K. Coker ◽  
E. Abe ◽  
R. L. Jilka ◽  
S. C. Manolagas
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Bone Morphogenetic Protein ◽  
Colony Formation ◽  
Tartrate Resistant Acid Phosphatase ◽  
Murine Bone Marrow ◽  
Morphogenetic Protein ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Abstract Using primary murine bone marrow cell cultures, we demonstrate that inhibin suppresses osteoblastogenesis and osteoclastogenesis. In contrast, activin supports osteoblast formation (by alkaline phosphatase-positive and mineralized colony formation); and activin also stimulates osteoclast formation (as measured by staining tartrate-resistant acid phosphatase-positive multinucleated cells). Inhibin, the activin antagonist follistatin, and the bone morphogenetic protein antagonist noggin can all suppress endogenous activin accumulation in bone marrow cultures. Associated with this decrease in activin is the loss of mineralized osteoblastic colony formation (colony forming unit-osteoblast; CFU-OB). However, exogenous activin administration, even in the presence of noggin, permits both alkaline phosphatase-positive and CFU-OB colony formation in vitro. In contrast, the stimulatory effects of locally produced activin on osteoblast and osteoclast development are not likely to be dominant over the suppressive effects of gonadally derived inhibin. The suppressive effect of inhibin is maintained in the presence of either activin or bone morphogenetic protein, suggesting the presence of a distinct inhibin-specific receptor. Taken together, the direct regulation of osteoblastogenesis and osteoclastogenesis by inhibin and activin in vitro suggest that changes in the inhibin/activin ratio detected by bone marrow cells, during the perimenopausal transition, contribute to altered cell differentiation and may be associated with the increased bone resorption observed at this time.

scholarly journals Adipocyte-Derived Exosomes Carrying Sonic Hedgehog Mediate M1 Macrophage Polarization-Induced Insulin Resistance via Ptch and PI3K Pathways

2018 ◽  
Vol 48 (4) ◽  
pp. 1416-1432 ◽  
Author(s):  
Ming Song ◽  
Lu Han ◽  
Fang-fang Chen ◽  
Di Wang ◽  
Feng Wang ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Western Blot ◽  
Sonic Hedgehog ◽  
High Glucose ◽  
Insulin Signalling ◽  
Macrophage Polarization ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
M1 Polarization ◽  
M1 Macrophage

Background/Aims: Adipocyte-derived exosomes (ADEs) stimulate the activation of macrophages and contribute to the development of insulin resistance. Sonic Hedgehog (Shh) is an exosome-carrying protein and stimulates macrophages to secrete inflammatory cytokines. However, the impact of ADEs carrying Shh on the pro-inflammatory activation of macrophages and consequently, adipocyte insulin resistance is unclear. Methods: 3T3-L1 adipocytes were cultured with high glucose and insulin to imitate the pathogeny of insulin resistance. ADEs were isolated from conditioned media of 3T3-L1 adipocytes via differential ultracentrifugation. We explored the role of ADEs carrying Shh in the polarization of macrophages by flow cytometry. Western blot and electrophoretic mobility shift assay (EMSA) were performed to determine the activation of Shh-mediated signalling pathways. The effects of ADE-treated macrophages on adipocyte insulin signalling were studied by Western blot. Results: We found that circulating Shh-positive exosomes were increased in type 2 diabetes patients. High glucose and insulin increased the secretion of Shh-positive ADEs. The ADEs carrying Shh induced pro-inflammatory or M1 polarization of bone marrow-derived macrophages (BMDM) and RAW 264.7 macrophages. Inhibitors of Ptch and PI3K blocked the M1 polarization induced by ADEs, which suggests that ADEs carrying Shh mediated M1 macrophage polarization through the Ptch/PI3K signalling pathway. ADE-treated RAW 264.7 macrophages were subsequently used to assess the effect on insulin signalling in adipocytes. Using a co-culture assay, we showed that both ADE-treated macrophages and exosomes from these macrophages could decrease the expression of insulin-resistant substrate-1 (IRS-1) and hormone-sensitive lipase (HSL) in adipocytes. Inhibitors of Ptch and PI3K blocked the down-regulation of IRS-1 and HSL induced by ADE-treated macrophages. Conclusion: Together, these data indicate that ADEs carrying Shh induce the M1 polarization of macrophages, which contributes to insulin resistance in adipocytes through the Ptch/PI3K pathway.

scholarly journals Altered stem cell (CFU-S) function following infection of hematopoietic cells with a virus carrying V-src

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 398-405 ◽  
Author(s):  
D Boettiger ◽  
TM Dexter
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Cells ◽  
Serial Passage ◽  
Self Renewal ◽  
Murine Bone Marrow ◽  
Bone Marrow Cultures ◽  
Normal Differentiation ◽  
High Level ◽  
Marrow Cultures

Abstract Long-term murine bone marrow cultures were used to support the growth and development of hematopoietic cells. After hematopoiesis was established, the cultures were infected with a recombinant murine amphotropic virus carrying the avian sarcoma virus src gene and the CFU- S kinetics were examined. The CFU-S from the src-infected cultures displayed a reduced seeding efficiency in the standard spleen colony assay. The self-renewal capacity of these CFU-S was tested by their ability to reestablish hematopoiesis when serially transplanted on irradiated bone marrow cultures and by serial passage in spleens of irradiated mice. In both tests, cells from the src-infected cultures exhibited an enhanced ability to sustain a high level of self-renewal. The other property of stem cells which may be measured is the probability of self-renewal at each cell division which dictates the distribution between stem cells and differentiated type progeny. CFU-S from the src-infected cultures had higher average probabilities of self- renewal and therefore reduced differentiation. These differences suggest that expression of src had indirectly or directly altered the normal differentiation program of the stem cells.

scholarly journals Altered stem cell (CFU-S) function following infection of hematopoietic cells with a virus carrying V-src

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 398-405
Author(s):  
D Boettiger ◽  
TM Dexter
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Cells ◽  
Serial Passage ◽  
Self Renewal ◽  
Murine Bone Marrow ◽  
Bone Marrow Cultures ◽  
Normal Differentiation ◽  
High Level ◽  
Marrow Cultures

Long-term murine bone marrow cultures were used to support the growth and development of hematopoietic cells. After hematopoiesis was established, the cultures were infected with a recombinant murine amphotropic virus carrying the avian sarcoma virus src gene and the CFU- S kinetics were examined. The CFU-S from the src-infected cultures displayed a reduced seeding efficiency in the standard spleen colony assay. The self-renewal capacity of these CFU-S was tested by their ability to reestablish hematopoiesis when serially transplanted on irradiated bone marrow cultures and by serial passage in spleens of irradiated mice. In both tests, cells from the src-infected cultures exhibited an enhanced ability to sustain a high level of self-renewal. The other property of stem cells which may be measured is the probability of self-renewal at each cell division which dictates the distribution between stem cells and differentiated type progeny. CFU-S from the src-infected cultures had higher average probabilities of self- renewal and therefore reduced differentiation. These differences suggest that expression of src had indirectly or directly altered the normal differentiation program of the stem cells.

Macrophage heterogeneity in bone marrow culture in vitro

1990 ◽  
Vol 95 (3) ◽  
pp. 481-485
Author(s):  
M. Mori ◽  
Y. Sadahira ◽  
S. Kawasaki ◽  
T. Hayashi ◽  
M. Awai
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Hematopoietic Cells ◽  
Bone Marrow Culture ◽  
Mouse Bone Marrow ◽  
Specific Monoclonal Antibody ◽  
Culture In Vitro ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Macrophages in mouse bone marrow cultures were investigated with macrophage-specific monoclonal antibody F4/80 and anti-Forssman glycosphingolipid (GSL) antibody, which was specific for macrophages in hematopoietic foci. Antibody F4/80 stained two types of cells, small macrophages and large flat macrophages associated with hematopoietic cells. The cytochemical and phagocytotic characteristics were similar between these two types of cells, but Forssman GSL was positive only for the large flat macrophages associated with hematopoietic cells. The data suggest that Forssman GSL positive macrophages, derived from resident bone marrow macrophages, play an important role in hematopoiesis and are clearly distinguished from small macrophages in vitro.

scholarly journals Cell types associated with fibronectin in long-term mouse bone marrow cultures.

1982 ◽  
Vol 30 (3) ◽  
pp. 235-244 ◽  
Author(s):  
U Reincke ◽  
P Hsieh ◽  
P Mauch ◽  
S Hellman ◽  
L B Chen
Keyword(s):  
Bone Marrow ◽  
Hematopoietic Cells ◽  
Cell Types ◽  
Mouse Bone Marrow ◽  
Cell Adherence ◽  
Fibronectin Matrix ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

The formation of fibronectin matrix was studied in long-term mouse bone marrow cultures. Stromal and hematopoietic cells were observed in situ under phase contrast optics and quantified according to their staining characteristics on smear preparations. Surface fibronectin was demonstrated by indirect immunofluorescence. While only stromal and no hematopoietic cells participated, various stromal cell types differed in their expression of cell surface fibronectin: Reticulum cells contributed the major portion of fibronectin matrix. Elongated, meshwork-forming histiocytes expressed some surface fibronectin, while the flattened, macrophagic histiocytes remained fibronectin negative. These findings were recapitulated during regeneration of scrape wounds in the adherent layers. Isolated fibronectin matrix did not support hematopoietic cell adherence or maintenance, although it had marked effects on stromal cells.

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