scholarly journals MiR-144-5p, an exosomal miRNA from bone marrow-derived macrophage in type 2 diabetes, impairs bone fracture healing via targeting Smad1

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Dong Zhang ◽  
Yifan Wu ◽  
Zonghuan Li ◽  
Hairen Chen ◽  
Siyuan Huang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Fracture Healing ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Fracture Repair ◽  
Osteogenic Potential ◽  
Patients With Diabetes ◽  
The Impact

Abstract Background Patients with diabetes have an increased risk of nonunion and delayed union of fractures. Macrophages have been shown as a key player in diabetic complications. However, it remains obscure how diabetic milieu affects macrophage-derived exosomes and its implications on osteogenic differentiation of BMSCs. In this study, we aim to define the impact of diabetic milieu on macrophage-derived exosomes, role of extracellular vesicles in intercellular communication with BMSCs, and subsequent effects on osteogenic differentiation and fracture repair. Results The osteogenic potential and the ability of fracture repair of exosomes derived from diabetic bone marrow-derived macrophages (dBMDM-exos) were revealed to be lower, as compared with non-diabetic bone marrow-derived macrophages (nBMDM-exos) in vitro and in vivo. Interestingly, miR-144-5p levels were sharply elevated in dBMDM-exos and it could be transferred into BMSCs to regulate bone regeneration by targeting Smad1. In addition, the adverse effects of dBMDM-exos on the osteogenic potential and the ability of fracture repair were reversed through the suppression of miR-144-5p inhibition in vitro and vivo. Conclusions The results demonstrated an important role of exosomal miR-144-5p in bone regeneration, offering insight into developing new strategy for the improvement of fracture healing in patients with diabetes mellitus. Graphic Abstract

Abstract 525: The Role of FHL2 in Mechanically Regulated Valve Interstitial Cell Osteogenic Differentiation

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Alan Y Lam ◽  
Jan-Hung Chen ◽  
Craig A Simmons
Keyword(s):  
Osteogenic Differentiation ◽  
Nuclear Translocation ◽  
Dominant Negative ◽  
Osteogenic Potential ◽  
Matrix Stiffness ◽  
Calcific Aortic Valve Disease ◽  
Regulatory Relationship ◽  
Sirna Knockdown

Introduction: Lesions in calcific aortic valve disease (CAVD) often include osteoblasts and occur preferentially in the natively stiffer fibrosa side of the valve, suggesting roles for osteogenesis and matrix stiffness in CAVD. Valve interstitial cells (VICs) have osteogenic potential that is modulated by matrix stiffness in vitro . Osteogenesis of mesenchymal stem cells is mediated by FHL2 and modulated by matrix stiffness via RhoA. However, the connection between FHL2 and RhoA and their roles in mechanically-regulated osteogenic differentiation of VICs and CAVD have not been established. Hypothesis: Matrix stiffness regulates RhoA-dependent FHL2 expression to direct VIC osteogenesis. Methods/Results: Primary porcine aortic VICs were grown on collagen-coated polyacrylamide gels of varying elastic moduli in vitro . With increasing substrate stiffness, VICs demonstrated increased RhoA activation by ELISA and increased FHL2 nuclear translocation by immunofluorescence and western blotting. Notably, significant increases were seen on 22 kPa substrates, a stiffness unique to the disease-prone fibrosa, compared to 11 kPa substrates (p<0.05). We investigated the regulatory relationship between RhoA and FHL2 in VICs by controlling RhoA levels with adenoviruses. With constitutively active RhoA, FHL2 nuclear translocation increased (p<0.06); in contrast, with dominant negative RhoA, FHL2 nuclear translocation decreased (p<0.05). The effect of RhoA levels on stiffness-dependent FHL2 activity was verified by knocking down RhoA levels pharmacologically (C3 toxin) which abrogated stiffness-dependent FHL2 nuclear translocation. Alkaline phosphatase activity as a marker of osteogenesis was highest on substrates with fibrosa-like stiffness and was reduced by FHL2 siRNA knockdown. Conclusions: Together, these results support the hypothesis that mechanically-regulated VIC osteogenic differentiation in vitro is mediated by FHL2 operating downstream of RhoA. New insights into the mechanotransduction mechanisms that regulate VIC osteogenesis may lead to novel strategies for treating CAVD.

Association of Dental Pulp Stem Cells and Ricinus Bone Compound in a Model of Bone Defect

2020 ◽  
Vol 3 (3) ◽  
pp. 267-278
Author(s):  
Alan Jesus ◽  
Adriano Jesus ◽  
Flávia Lima ◽  
Luiz Freitas ◽  
Cássio Meira ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Bone Defect ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Autogenous Bone ◽  
Control Group

Autogenous bone grafting is needed in some bone tissue defects; however, it causes secondary surgical wounds and morbidity. Tissue bioengineering may be an alternative approach for bone regeneration. Here we investigated the osteogenic potential of dental pulp stem cells from deciduous teeth (DPSC) in association with a Ricinus bone compound (RBC) in a model of bone defect. The influence of the biomaterial RBC on the proliferation and osteogenic differentiation of DPSC was assessed in vitro by MTT metabolism and alizarin red staining, respectively. The morphologic analysis was performed using the optic and scanning electron (SEM) microscopies. For the in vivo study, 54 Wistar rats submitted to calvarial defects were filled with RBC or RBC+DPSC. A control group had the defects filled only with blood clots. Analyses were performed 15, 30 and 60 days after treatment using digital radiography, optical microscopy, SEM and chemical analysis by electron dispersive spectroscopy. The Ricinus bone compound (RBC) did not inhibit the osteogenic differentiation in vitro. No spontaneous regeneration was observed in the control group. The area of the calvarial defect of the RBC+DPSC group showed greater radiopacity on day 15. The RBC presented no reabsorption, was biocompatible and showed osteointegration, working as a mechanical filling. Only sparse ossification areas were found and those were larger and more developed on the RBC+DPSC group when compared to animals treated only with RBC. RBC in association with DPSC is a promising combination for applications in bone regeneration.  

scholarly journals Role of Fzd6 in Regulating the Osteogenic Differentiation of Adipose-derived Stem Cells in Osteoporosis

2021 ◽  
Author(s):  
Tianli Wu ◽  
Zhihao Yao ◽  
Gang Tao ◽  
Fangzhi Lou ◽  
Hui Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Wnt Signaling Pathway ◽  
Osteogenic Potential ◽  
Adipose Derived Stem Cells ◽  
Alizarin Red ◽  
Differentiation Potential

Abstract Objective: Although it has been demonstrated that adipose-derived stem cells (ASCs) from osteoporosis mice (OP-ASCs) exhibit impaired osteogenic differentiation potential, the molecular mechanism has not yet been elucidated. We found that Fzd6 was decreased in OP-ASCs compared with ASCs. This study investigates the effects and underlying mechanisms of Fzd6 in the osteogenic potential of OP-ASCs. Methods: Fzd6 expression in ASCs and OP-ASCs was measured by PCR gene chip. Fzd6 overexpression and silencing lentiviruses were used to evaluate the role of Fzd6 in the osteogenic differentiation of OP-ASCs. Real-time PCR (qPCR) and western blotting (WB) was performed to detect the expression of Fzd6 and bone-related molecules, including runt-related transcription factor 2 (Runx2) and osteopontin (Opn). Alizarin red staining and Alkaline phosphatase (ALP) staining was performed following osteogenic induction. Microscopic CT (Micro-CT), hematoxylin and eosin staining (H&E) staining, and Masson staining were used to assess the role of Fzd6 in osteogenic differentiation of osteoporosis (OP) mice in vivo.Results: Expression of Fzd6 was decreased significantly in OP-ASCs. Fzd6 silencing down-regulated the osteogenic ability of OP-ASCs in vitro. Overexpression of Fzd6 rescued the impaired osteogenic capacity in OP-ASCs in vitro. We obtained similar results in vivo.Conclusions: Fzd6 plays an important role in regulating the osteogenic ability of OP-ASCs both in vivo and in vitro. Overexpression of Fzd6 associated with the Wnt signaling pathway promotes the osteogenic ability of OP-ASCs, which provides new insights for the prevention and treatment of OP.

A Tumor Suppressor microRNA Defines the Leukemic Hierarchy in Acute Myeloid Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3653-3653
Author(s):  
Edith Schneider ◽  
Anna Staffas ◽  
Linda Roehner ◽  
Christian Buske ◽  
Lars Bullinger ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Tumor Suppressor ◽  
Murine Bone Marrow ◽  
Progression Model ◽  
Step Model ◽  
Total Bone Marrow ◽  
The Impact

Abstract In order to identify functionally relevant microRNAs (miRNA) in AML, we profiled global miRNA expression in a murine AML progression model based on Hoxa9 and Meis1 overexpression. We found miR-155 and miR-708 as the most significantly upregulated miRNA species in leukemic Hoxa9/Meis1 cells compared to the preleukemic Hoxa9/ctrl cells (both p<0.01). Subsequent analysis of various AML subtypes (CN-AML, t(11q23), t(8;21), t(15;17), n=38) showed significantly elevated levels of miR-155 and miR-708 in all tested samples (n=5-10) compared to total bone marrow from healthy donors, indicating potential oncogenic roles for these miRNAs. We further investigated, in vivo, the role of miR-155 and miR-708 in AML by retroviral overexpression with Hoxa9 in murine bone marrow (mbm) cells followed by syngeneic transplantation. Overexpression of miR-155 in conjunction with Hoxa9 (Hoxa9/miR-155) caused a significantly accelerated onset of a myelomonocytic leukemia within 93 days (survival 93±23 days, p<0.0001), but still a less aggressive course of disease compared to mice transplanted with Hoxa9/Meis1 (survival 36±4 days, p<0.0001). We then hypothesized that the combination of miR-155 and miR-708 could further replace the oncogenic potential of Meis1. Therefore, mbm cells were retrovirally transduced with Hoxa9, miR-155 and miR-708 (Hoxa9/155/708) or Hoxa9 and miR-708 (Hoxa9/miR-708) and functionally analyzed in vitro and in vivo. To our surprise, miR-708 abrogated the leukemogenic effect of Hoxa9, alone or in combination with Hoxa9/miR-155 in vivo (p=0.0117, p<0.0001, the mice were sacrificed after 197 days), with little or no engraftment. To understand why miR-708, a potent tumor suppressor miRNA, is upregulated in the highly aggressive Hoxa9/Meis1 AML cells, we first showed in vivo that miR-708 does not impair the development of AML by Hoxa9/Meis1 cells. In order to explore the role of miR-708 in leukemia initiating cells (LIC), we FACS-sorted subpopulations enriched for LIC based on c-kit, Mac-1 and Gr-1 expression within the Hoxa9/Meis1 cells. Of the three subpopulations (c-kit+ Gr-1- Mac-1-, c-kit+ Gr-1+ Mac-1+, c-kit- Gr-1+ Mac-1+) the transplanted c-kit+ Gr-1- Mac-1- cells caused a significantly shorter survival (survival 36 days, range: 32-37d) compared to the other sorted subpopulations (p=0.0072 and p=0.0021, respectively), whereas the c-kit- Gr-1+ Mac-1+ subpopulation exhibited the longest survival (survival 44 days, range: 42-51d). Based on these findings, we measured miR-708 and miR-155 levels in the sorted Hoxa9/Meis1 subpopulations by qRT-PCR. Strikingly, no difference compared to bulk cells was detected for miR-155, highlighting its role as an oncogenic driver in all subpopulations. In contrast, miR-708 showed a distinct expression pattern with lower expression in the LIC-enriched c-kit+ Gr-1- Mac-1- subpopulation compared to bulk (p=0.032), allowing discernment of the LIC enriched population and highlighting its role as an orchestrator of the leukemic hierarchy. Adapted from these findings, we developed an AML 4-step model (Hoxa9/miR-708, Hoxa9/ctrl, Hoxa9/miR-155, Hoxa9/Meis1) based on four immortalized Hoxa9 based cell lines with completely different in vivo properties ranging from no engraftment to induction of highly aggressive AML. Considering the central role of Hoxa9/Meis1 in AML, we demonstrate, for the first time, the concept of a tumor suppressor miRNA that stratifies the leukemic hierarchy driven by an oncogenic miRNA. Moreover, we present a new AML 4-step model as a novel tool that will allow detailed investigation of the impact of new oncogenes at every step of leukemogenesis. Disclosures Buske: CELLTRION, Inc.: Consultancy, Honoraria.

scholarly journals Strontium Promotes the Proliferation and Osteogenic Differentiation of Human Placental Decidual Basalis- and Bone Marrow-Derived MSCs in a Dose-Dependent Manner

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Yi-Zhou Huang ◽  
Cheng-Guang Wu ◽  
Hui-Qi Xie ◽  
Zhao-Yang Li ◽  
Antonietta Silini ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Osteogenic Differentiation ◽  
Early Stage ◽  
Cell Types ◽  
Osteogenic Potential ◽  
Dependent Manner ◽  
Gene Expressions ◽  
Dose Dependent ◽  
Osteogenic Gene

The osteogenic potential of mesenchymal stromal cells (MSCs) varies among different tissue sources. Strontium enhances the osteogenic differentiation of bone marrow-derived MSCs (BM-MSCs), but whether it exerts similar effects on placental decidual basalis-derived MSCs (PDB-MSCs) remains unknown. Here, we compared the influence of strontium on the proliferation and osteogenic differentiation of human PDB- and BM-MSCs in vitro. We found that 1 mM and 10 mM strontium, but not 0.1 mM strontium, evidently promoted the proliferation of human PDB- and BM-MSCs. These doses of strontium showed a comparable alkaline phosphatase activity in both cell types, but their osteogenic gene expressions were promoted in a dose-dependent manner. Strontium at doses of 0.1 mM and 1 mM elevated several osteogenic gene expressions of PDB-MSCs, but not those of BM-MSCs at an early stage. Nevertheless, they failed to enhance the mineralization of either cell type. By contrast, 10 mM strontium facilitated the osteogenic gene expression as well as the mineralization of human PDB- and BM-MSCs. Collectively, this study demonstrated that human PDB- and BM-MSCs shared a great similarity in response to strontium, which promoted their proliferation and osteogenic differentiation in a dose-dependent manner.

Apigenin promotes osteogenic differentiation of mesenchymal stem cells and accelerates bone fracture healing via activating Wnt/β-catenin signaling

2021 ◽  
Author(s):  
Fei-fei Pan ◽  
Jiang Shao ◽  
Chuan-jian Shi ◽  
Zhi-peng Li ◽  
Wei-ming Fu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Fracture Healing ◽  
Osteogenic Differentiation ◽  
Bone Fracture ◽  
Fruits And Vegetables ◽  
Target Genes ◽  
Fracture Repair

Apigenin (API), a natural plant flavone, is abundantly found in common fruits and vegetables. As a bioactive flavonoid, API exhibits several activities including anti-proliferation and anti-inflammation. A recent study showed that API could retard osteoporosis progress, indicating its role in the skeletal system. However, the detailed function and mechanism remain obscure. In the present study, API was found to promote osteogenic differentiation of mesenchymal stem cells (MSCs). And further investigation showed that API could enhance the expression of the critical transcription factor β-catenin and several downstream target genes of Wnt signaling, thus activated Wnt/β-catenin signaling. Using a rat femoral fracture model, API was found to improve new bone formation and accelerate fracture healing in vivo. In conclusion, our data demonstrated that API could promote osteogenesis in vitro and facilitate the fracture healing in vivo via activating Wnt/β-catenin signaling, indicating that API may be a promising therapeutic candidate for bone fracture repair.

scholarly journals Bone-Forming Peptide-4 Induces Osteogenic Differentiation and VEGF Expression on Multipotent Bone Marrow Stromal Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Mi Eun Kim ◽  
Jong Keun Seon ◽  
Ju Yeon Kang ◽  
Taek Rim Yoon ◽  
Jun Sik Lee ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Formation ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Bone Repair ◽  
Fracture Repair ◽  
Marrow Stromal Cells ◽  
Dependent Manner

Bone morphogenetic proteins (BMPs) have been widely used as treatment for bone repair. However, clinical trials on fracture repair have challenged the effectiveness of BMPs and suggested that delivery of multipotent bone marrow stromal cells (BMSCs) might be beneficial. During bone remodeling and bone fracture repair, multipotent BMSCs differentiate into osteoblasts or chondrocytes to stimulate bone formation and regeneration. Stem cell-based therapies provide a promising approach for bone formation. Extensive research has attempted to develop adjuvants as specific stimulators of bone formation for therapeutic use in patients with bone resorption. We previously reported for the first time bone-forming peptides (BFPs) that induce osteogenesis and bone formation. BFPs are also a promising osteogenic factor for prompting bone regeneration and formation. Thus, the aim of the present study was to investigate the underlying mechanism of a new BFP-4 (FFKATEVHFRSIRST) in osteogenic differentiation and bone formation. This study reports that BFP-4 induces stronger osteogenic differentiation of BMSCs than BMP-7. BFP-4 also induces ALP activity, calcium concentration, and osteogenic factors (Runx2 and osteocalcin) in a dose dependent manner in BMSCs. Therefore, these results indicate that BFP-4 can induce osteogenic differentiation and bone formation. Thus, treatment of multipotent BMSCs with BFP-4 enhanced osteoblastic differentiation and displayed greater bone-forming ability than BMP-7 treatment. These results suggest that BFP-4-stimulated cell therapy may be an efficient and cost-effective complement to BMP-7-based clinical therapy for bone regeneration and formation.

scholarly journals Impact of Donor Age on the Osteogenic Supportive Capacity of Mesenchymal Stromal Cell-Derived Extracellular Matrix

2021 ◽  
Vol 9 ◽  
Author(s):  
Marta S. Carvalho ◽  
Laura Alves ◽  
Isabel Bogalho ◽  
Joaquim M. S. Cabral ◽  
Cláudia L. da Silva
Keyword(s):  
Extracellular Matrix ◽  
Osteogenic Differentiation ◽  
Bone Fractures ◽  
Therapeutic Option ◽  
Osteogenic Potential ◽  
Donor Age ◽  
Gene Markers ◽  
Age Related ◽  
The Impact

Mesenchymal stromal cells (MSC) have been proposed as an emerging cell-based therapeutic option for regenerative medicine applications as these cells can promote tissue and organ repair. In particular, MSC have been applied for the treatment of bone fractures. However, the healing capacity of these fractures is often compromised by patient’s age. Therefore, considering the use of autologous MSC, we evaluated the impact of donor age on the osteogenic potential of bone marrow (BM)-derived MSC. MSC from older patients (60 and 80 years old) demonstrated impaired proliferative and osteogenic capacities compared to MSC isolated from younger patients (30 and 45 years old), suggesting that aging potentially changes the quantity and quality of MSC. Moreover, in this study, we investigated the capacity of the microenvironment [i.e., extracellular matrix (ECM)] to rescue the impaired proliferative and osteogenic potential of aged MSC. In this context, we aimed to understand if BM MSC features could be modulated by exposure to an ECM derived from cells obtained from young or old donors. When aged MSC were cultured on decellularized ECM derived from young MSC, their in vitro proliferative and osteogenic capacities were enhanced, which did not happen when cultured on old ECM. Our results suggest that the microenvironment, specifically the ECM, plays a crucial role in the quality (assessed in terms of osteogenic differentiation capacity) and quantity of MSC. Specifically, the aging of ECM is determinant of osteogenic differentiation of MSC. In fact, old MSC maintained on a young ECM produced higher amounts of extracellularly deposited calcium (9.10 ± 0.22 vs. 4.69 ± 1.41 μg.μl–1.10–7 cells for young ECM and old ECM, respectively) and up-regulated the expression of osteogenic gene markers such as Runx2 and OPN. Cell rejuvenation by exposure to a functional ECM might be a valuable clinical strategy to overcome the age-related decline in the osteogenic potential of MSC by recapitulating a younger microenvironment, attenuating the effects of aging on the stem cell niche. Overall, this study provides new insights on the osteogenic potential of MSC during aging and opens new possibilities for developing clinical strategies for elderly patients with limited bone formation capacity who currently lack effective treatments.

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