Bone marrow mechanotransduction in porcine explants alters kinase activation and enhances trabecular bone formation in the absence of osteocyte signaling

Bone ◽  
2018 ◽  
Vol 107 ◽  
pp. 78-87 ◽  
Author(s):  
Kimberly J. Curtis ◽  
Thomas R. Coughlin ◽  
Devon E. Mason ◽  
Joel D. Boerckel ◽  
Glen L. Niebur
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Trabecular Bone ◽  
Kinase Activation

scholarly journals Antiosteoporotic Activity ofDioscorea alataL. cv. Phyto through Driving Mesenchymal Stem Cells Differentiation for Bone Formation

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Kang-Yung Peng ◽  
Lin-Yea Horng ◽  
Hui-Ching Sung ◽  
Hui-Chuan Huang ◽  
Rong-Tsun Wu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Bone Formation ◽  
Trabecular Bone ◽  
Nodule Formation ◽  
Mineral Density ◽  
Osteoporosis Therapy ◽  
Trabecular Bone Mineral Density ◽  
Bone Nodule ◽  
Bone Nodule Formation

The aim of this study was to evaluate the effect of an ethanol extract of the rhizomes ofDioscorea alataL. cv. Phyto, Dispo85E, on bone formation and to investigate the mechanisms involved. Our results showed that Dispo85E increased the activity of alkaline phosphatase (ALP) and bone nodule formation in primary bone marrow cultures. In addition, Dispo85E stimulated pluripotent C3H10T1/2 stem cells to differentiate into osteoblasts rather than adipocytes. Ourin vivodata indicated that Dispo85E promotes osteoblastogenesis by increasing ALP activity and bone nodule formation in both intact and ovariectomized (OVX) mice. Microcomputed tomography (μCT) analysis also showed that Dispo85E ameliorates the deterioration of trabecular bone mineral density (tBMD), trabecular bone volume/total volume (BV/TV), and trabecular bone number (Tb.N) in OVX mice. Our results suggested that Dispo85E is a botanical drug with a novel mechanism that drives the lineage-specific differentiation of bone marrow stromal cells and is a candidate drug for osteoporosis therapy.

scholarly journals Impaired Osteoblastic Differentiation of MSCs Suppresses Normal Hematopoiesis in MDS

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 17-18
Author(s):  
Yasutaka Hayashi ◽  
Kimihito Cojin Kawabata ◽  
Yosuke Tanaka ◽  
Yasufumi Uehara ◽  
Shigeru Kiryu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Bone Formation ◽  
Hematopoietic Stem Cells ◽  
Trabecular Bone ◽  
Bone Marrow Failure ◽  
Bone Volume ◽  
Marker Genes ◽  
Double Labeling ◽  
Hematopoietic Stem

Myelodysplastic syndromes (MDS) is a clonal disorder of hematopoietic stem cells (HSCs) characterized by clonal hematopoietic stem cells (HSCs) with cytopenia, morphological abnormalities, genetic alteration, ineffective normal hematopoiesis, and frequent progression to AML. It has long remained unresolved how MDS cells, which are less proliferative, inhibit normal hematopoiesis and eventually come to dominate the bone marrow space. Despite several studies of mesenchymal stem cells (MSCs), one of the principal components of HSC niche supporting normal hematopoiesis, the molecular mechanisms underlying this process remain unclear. In this study, we examined the mechanism by which less-proliferative MDS cells outcompete normal hematopoiesis through the effects on MSCs using serially transplantable Abcg2-induced MDS/AML model we recently generated. The recipient-derived normal BM cells displayed a considerably lower colony output with markedly decreased numbers of the hematopoietic stem progenitor cells (HSPCs) . However, there were no direct effects on the colony-forming ability of the recipient HSPCs co-cultured with MDS/AML cells, indicating that MDS/AML cells inhibited hematopoiesis through alteration of bone marrow microenvironment, such as MSCs, rather than direct interaction between normal and malignant HSCs. We next analyzed histological features of BM specimens. Interestingly, bone sections from the MDS/AML mice showed a reduced trabecular bone and narrowed growth plates. Moreover, micro computed tomography (micro-CT) analysis of the femora showed a significant reduction of the trabecular bone volume in the recipient mice transplanted with the MDS/AML BM cells. We detected decreased bone formation based on the calcein double labeling, but unchanged numbers of the TRAP-positive mononuclear or multinucleated (osteoclastic) cells in the MDS/AML samples, suggesting that the reduced bone volume was caused by suppressed bone formation. The impaired bone formation was also observed in the human MDS patients in terms of lower bone volume and decreased expression of BGLAP, one of osteogenic markers. In line with the above findings, single cell qRT-PCR analyses of mouse MSCs displayed downregulation of a line of osteolineage markers, indicating that MDS/AML cells suppress bone formation through inhibiting osteolineage differentiation of MSCs. Based on the findings, we next examined if re-induction of osteolineage differentiation of the MDS/AML-derived MSCs could rescue the potential of MSCs to support normal hematopoiesis. Importantly, the number of colony-forming cells (CFCs) was significantly restored by inducing differentiation of MDS/AML-derived MSCs toward osteogenic lineage both in vitro and in vivo. These results indicate that the impairment of osteolineage differentiation is the principal cause for an impaired normal hematopoiesis in MDS/AML, and that restoring the supportive niche will be a potential therapeutic option. Since extracellular vesicles (EVs) derived from MDS/AML cells are critical mediators of intercellular communication, we examined the molecular mechanism underlying the dysfunction of MSCs via EVs. As expected, EVs from MDS/AML cells were incorporated into the normal MSCs where osteolineage marker genes were clearly downregulated, and the number of CFCs significantly decreased in the HSPCs co-cultured with MSCs treated by the MDS/AML-derived EVs. Moreover, by comprehensively analyzing microRNAs (miRNAs) enriched in EVs derived from MDS/AML cells, we identified several miRNAs that impaired the differentiation of normal MSCs. These results suggested that miRNAs in EVs derived from MDS/AML cells disrupted the hematopoietic supporting niche through suppressing an osteolineage differentiation of MSCs. Here we uncover a heretofore unrecognized mechanism of bone marrow failure in MDS via the impairment of osteolineage differentiation in MSCs. EVs from MDS cells will be an attractive therapeutic target to restore the supportive niches, such as MSCs, for the remaining normal HSCs. Figure Disclosures No relevant conflicts of interest to declare.

scholarly journals BMP4 Gene Therapy Inhibits Myeloma Tumor Growth, but Has a Negative Impact on Bone

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1928-1928 ◽  
Author(s):  
Toril Holien ◽  
Marita Westhrin ◽  
Siv Helen Moen ◽  
Muhammad Zahoor ◽  
Glenn Buene ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Multiple Myeloma ◽  
Bone Marrow ◽  
Bone Formation ◽  
Tumor Growth ◽  
Trabecular Bone ◽  
Plasma Cells ◽  
Bmp Signaling ◽  
Myeloma Cells ◽  
Post Test

Abstract TH and MW contributed equally to this work. Background Multiple myeloma is caused by an accumulation of malignant plasma cells in the bone marrow. Myeloma is characterized by an osteolytic bone disease, caused by increased bone degradation and reduced bone formation. Bone morphogenetic proteins (BMPs) are members of the transforming growth factor (TGF)-β superfamily. BMP-signaling is important for both pre- and postnatal bone formation. Additionally, several BMPs induce growth arrest and apoptosis in myeloma cells. Thus, increasing BMP-signaling in myeloma patients may reduce tumor growth and restore bone formation. We therefore explored BMP4 gene therapy in a human-mouse model of multiple myeloma. Methods Calcium phosphate scaffolds with human mesenchymal stromal cells (MSCs) were implanted in RAG2-/-GC-/- mice and the MSCs were left to differentiate in vivo for 8 weeks to create a humanized bone microenvironment. Then, adeno-associated virus (AAV), AAV8-BMP4, which has tropism for liver cells and expresses murine Bmp4 under the control of the liver specific human α1-antitrypsin (hAAT1) promoter, were administered by tail-vein injection. Empty viral vectors, AAV8-CTRL, were used for the control group. After 2 weeks, when BMP4 was detectable in circulation, we injected fluorescently labelled KJON myeloma cells in 3 out of 4 scaffolds in each mouse. The KJON cells are hyperdiploid, have a relatively slow growth rate and rely on interleukin (IL)-6 supplementation in the absence of a supporting microenvironment, thus resembling primary human myeloma cells. Tumor growth was examined by weekly imaging until end-point, 6 weeks after tumor cell injection. Results At end-point, serum levels of BMP4 in AAV8-BMP4 mice were in the range of 50-200 ng/mL, but not detectable in AAV8-CTRL mice. Strikingly, tumor growth as quantified by imaging was significantly reduced in AAV8-BMP4 mice compared with the AAV8-CTRL mice (p<0.01, 2-way ANOVA, Bonferroni post-test), suggesting that high levels of circulating BMP4 reduced tumor growth. Malignant plasma cells were not detected in the murine bone marrow or spleen as examined by imaging and flow cytometry, suggesting that the myeloma cells were confined to the humanized scaffolds. Myeloma cells isolated from tumors from both AAV8-BMP4 and AAV8-CTRL mice were still sensitive to BMP4 treatment in vitro, indicating that they did not acquire resistance to BMP4 during the experiment. We hypothesized that increased circulating BMP4 would also be beneficial for bone in this model. Human bone is generated on the scaffolds by osteoblasts that differentiate from human MSCs seeded on the scaffolds before implantation. In this model, myeloma cells inhibit osteoblast differentiation and bone formation. Indeed, in AAV8-CTRL mice we found less bone in tumor cell-containing scaffolds compared with scaffolds without tumor cells (p<0.05, Kruskal-Wallis with Dunn's post-test). However, bone formation was not increased in tumor-containing scaffolds of AAV8-BMP4 mice. To delineate the effects of BMP4 overexpression on bone per se, without direct influence from the cancer cells, we examined the femurs by μCT. Surprisingly, the AAV8-BMP4 mice had significantly reduced trabecular bone volume (p=0.017), trabecular numbers (p=0.016), as well as significantly increased trabecular separation (p<0.001) compared with the AAV8-CTRL mice. Thus, high levels of circulating BMP4 seemed to inhibit murine trabecular bone formation. There was no difference in cortical bone parameters between the two groups. Conclusion Taken together, BMP4 gene therapy inhibited myeloma tumor growth, but also reduced trabecular bone formation in mice. Care should be taken when considering BMP4 as a therapeutic agent. If other BMPs that are also potent inhibitors of multiple myeloma cell survival and proliferation will have similar impact on bone remains to be investigated. Disclosures No relevant conflicts of interest to declare.

scholarly journals RESPON TULANG FEMUR TIKUS OSTEOPOROSIS YANG MENGKONSUMSI CALCITRIOL

2018 ◽  
Vol 36 (1) ◽  
pp. 1
Author(s):  
Hartiningsih Hartiningsih ◽  
Devita Anggraeni
Keyword(s):  
Bone Marrow ◽  
Bone Resorption ◽  
Bone Formation ◽  
Trabecular Bone ◽  
Distal Femur ◽  
Histopathological Examination ◽  
Diet Group ◽  
Ovariectomized Rats ◽  
Standard Diet ◽  
The Right

Calcitriol supplementation in ovariectomized rats decreased bone resorption and increased bone formation, however, it depend on dose. The objective of the research was to study the response of femur bone in osteoporosis rats consuming calcitriol. Thirty female Wistar rats at 8 weeks of age were randomly divided into six groups (sham operated rats as normal control rats/group N and NK, ovariectomized control rats/group Ov and OvK, ovariectomized rats/group OvDand OvE) of five each. All rats were fed standard diet for 8 weeks. At 16 weeks  of age, group N and Ov  were euthanized, the right femur were taken for histopathological  examination. Group NK and OvK were fed a standard diet, group OvD was fed a standard diet +40ng calcitriol;  and group OvE was fed a standard diet+25µg ethynil ethyl estradiol. Treatments were done for six weeks. At the end of study, blood samples were taken from plexus orbitalis medialis for estrogen analysis. All rats were euthanized using ketamine10% and xylazine 2%. Right femur was taken for histopathological examination  using hematoxylin and eosin stain, and immunohistochemistry using monoclonal antibody anti TRAP5b which was detected with streptavidin-biotin.         The results showed that estradiol level of the rats in group OvD was not significantly different compared with the rats in OvK group, however, it was significanly lower compared to the rats in group OvE. Histopathologic figure of right distal femur metaphysis in group OvD was shown lesser adipocyte in the bone marrow and more trabecular bone speculum compared to group  OvK, however, there was more adipocyte in the bone marrow and lesser trabecular bone speculum compared to group OvE. Immunohistochemistry of distal femur metaphysis in group OvD and OvE were revealed tartrate resistant alkaline phosphatase 5b (TRAP5b) expression in trabecular bone, which was located in bone marrow space and trabecular speculum surface as well. Based on the results, it can be concluded that calcitriol 40ng/day supplementation in osteoporosis rats for 6 weeks decreased bone resorption and increased bone formation distal femur metaphysis.

Author response for "Piezo1 Inactivation in Chondrocytes Impairs Trabecular Bone Formation"

2020 ◽  
Author(s):  
Gretl Hendrickx ◽  
Verena Fischer ◽  
Astrid Liedert ◽  
Simon von Kroge ◽  
Melanie Haffner‐Luntzer ◽  
...  
Keyword(s):  
Bone Formation ◽  
Trabecular Bone ◽  
Author Response

scholarly journals Topical co‐administration of zoledronate with recombinant human bone morphogenetic protein-2 can induce and maintain bone formation in the bone marrow environment

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Hideki Ueyama ◽  
Yoichi Ohta ◽  
Yuuki Imai ◽  
Akinobu Suzuki ◽  
Ryo Sugama ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Bone Structure ◽  
Precursor Cells ◽  
The Other ◽  
Bone Morphogenetic Protein 2 ◽  
New Bone Formation ◽  
Micro Computed Tomography ◽  
Mineral Density ◽  
Left Femur

Abstract Background Bone morphogenetic proteins (BMPs) induce osteogenesis in various environments. However, when BMPs are used alone in the bone marrow environment, the maintenance of new bone formation is difficult owing to vigorous bone resorption. This is because BMPs stimulate the differentiation of not only osteoblast precursor cells but also osteoclast precursor cells. The present study aimed to induce and maintain new bone formation using the topical co-administration of recombinant human BMP-2 (rh-BMP-2) and zoledronate (ZOL) on beta-tricalcium phosphate (β-TCP) composite. Methods β-TCP columns were impregnated with both rh-BMP-2 (30 µg) and ZOL (5 µg), rh-BMP-2 alone, or ZOL alone, and implanted into the left femur canal of New Zealand white rabbits (n = 56). The implanted β-TCP columns were harvested and evaluated at 3 and 6 weeks after implantation. These harvested β-TCP columns were evaluated radiologically using plane radiograph, and histologically using haematoxylin/eosin (H&E) and Masson’s trichrome (MT) staining. In addition, micro-computed tomography (CT) was performed for qualitative analysis of bone formation in each group (n = 7). Results Tissue sections stained with H&E and MT dyes revealed that new bone formation inside the β-TCP composite was significantly greater in those impregnated with both rh-BMP-2 and ZOL than in those from the other experimental groups at 3 and 6 weeks after implantations (p < 0.05). Micro-CT data also demonstrated that the bone volume and the bone mineral density inside the β-TCP columns were significantly greater in those impregnated with both rh-BMP-2 and ZOL than in those from the other experimental groups at 3 and 6 weeks after implantations (p < 0.05). Conclusions The topical co-administration of both rh-BMP-2 and ZOL on β-TCP composite promoted and maintained newly formed bone structure in the bone marrow environment.

scholarly journals Circular RNA AFF4 modulates osteogenic differentiation in BM-MSCs by activating SMAD1/5 pathway through miR-135a-5p/FNDC5/Irisin axis

2021 ◽  
Vol 12 (7) ◽  
Author(s):  
Chao Liu ◽  
An-Song Liu ◽  
Da Zhong ◽  
Cheng-Gong Wang ◽  
Mi Yu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Osteogenic Differentiation ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Regulatory System ◽  
Circular Rna ◽  
Luciferase Reporter ◽  
Integrin Αv

AbstractBone marrow-derived mesenchymal stem cells (BM-MSCs), the common progenitor cells of adipocytes and osteoblasts, have been recognized as the key mediator during bone formation. Herein, our study aim to investigate molecular mechanisms underlying circular RNA (circRNA) AFF4 (circ_AFF4)-regulated BM-MSCs osteogenesis. BM-MSCs were characterized by FACS, ARS, and ALP staining. Expression patterns of circ_AFF4, miR-135a-5p, FNDC5/Irisin, SMAD1/5, and osteogenesis markers, including ALP, BMP4, RUNX2, Spp1, and Colla1 were detected by qRT-PCR, western blot, or immunofluorescence staining, respectively. Interactions between circ_AFF4 and miR-135a-5p, FNDC5, and miR-135a-5p were analyzed using web tools including TargetScan, miRanda, and miRDB, and further confirmed by luciferase reporter assay and RNA pull-down. Complex formation between Irisin and Integrin αV was verified by Co-immunoprecipitation. To further verify the functional role of circ_AFF4 in vivo during bone formation, we conducted animal experiments harboring circ_AFF4 knockdown, and born samples were evaluated by immunohistochemistry, hematoxylin and eosin, and Masson staining. Circ_AFF4 was upregulated upon osteogenic differentiation induction in BM-MSCs, and miR-135a-5p expression declined as differentiation proceeds. Circ_AFF4 knockdown significantly inhibited osteogenesis potential in BM-MSCs. Circ_AFF4 stimulated FNDC5/Irisin expression through complementary binding to its downstream target molecule miR-135a-5p. Irisin formed an intermolecular complex with Integrin αV and activated the SMAD1/5 pathway during osteogenic differentiation. Our work revealed that circ_AFF4, acting as a sponge of miR-135a-5p, triggers the promotion of FNDC5/Irisin via activating the SMAD1/5 pathway to induce osteogenic differentiation in BM-MSCs. These findings gained a deeper insight into the circRNA-miRNA regulatory system in the bone marrow microenvironment and may improve our understanding of bone formation-related diseases at physiological and pathological levels.

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