Expression Of Bone Morphogenetic Protein Research Articles

Hesperidin Exherts Antiglaucoma Effects by Activating Overexpression of BMP4 Signaling and Management of Retinal Ganglionic Cells Degeneration

We investigated the effect of hesperidin on bone morphogenetic protein 4 signaling, which is involved in preventing retinal ganglion cell degeneration in rats with glaucoma. We investigated the therapeutic efficacy of hesperidin as an enhancer of the bone morphogenetic protein 4 signaling pathway at three dose levels (50, 100, and 200 mg/kg) using a rat glaucoma model produced by injecting conjunctival fibroblasts intracamerally into the eyes of the animals. Bone morphogenetic protein 4 was stimulated with bilateral intravitreal injections of 2 mL adeno-associated virus vectors cloned with bone morphogenetic protein 4 primers into the right eye. The intraocular pressure, expression of bone morphogenetic protein 4 and Smad 4, and the distribution of the retinal ganglion cells count were performed a week following the onset of glaucoma. When compared to the model group, the retinal ganglion cells survival rate was increased proportionately in the hesperidin group (P < 0.01). A significant increase in bone morphogenetic protein 4 messenger ribonucleic acid expression was observed in the hesperidin group (P < 0.01) compared to the model group. The hesperidin group showed similar positive results (P < 0.01) with intraocular pressure, Western blot analysis, retinal explant culture, and histological analysis. Further, the model group showed a decline in retinal ganglion cells survival rate (P < 0.01) compared to the control group, and similar trends were noticed in intraocular pressure, Western blot analyses, retinal explant culture, and histological analysis. According to our findings, hesperidin increases the activity of the bone morphogenetic protein 4 pathway, which in turn increases the survival of retinal ganglion cells and the function of the eyes in an experimental glaucoma model.

Secretion of BMP-2 by tumor-associated macrophages (TAM) promotes microcalcifications in breast cancer

IntroductionBreast microcalcifications is a characteristic feature in diagnostic imaging and a prognostic factor of breast cancer. However, the underlying mechanisms of breast microcalcifications formation are not fully understood. Previous studies have shown that upregulation of bone morphogenetic protein 2 (BMP-2) is associated with the occurrence of microcalcifications and tumor-associated macrophages (TAMs) in the tumor microenvironment can secrete BMP-2. The aim of this study is to elucidate the role of secretion of BMP-2 by TAMs in promoting microcalcifications of breast cancer through immunohistochemical staining and co-culturing of breast cancer cells with TAMs.MethodsA total of 272 patients diagnosed with primary invasive breast cancer from January 2010 to January 2012 in the First Hospital of China Medical University were included in this study. Immunohistochemical staining of CD68 (marker of entire macrophages), CD168 (marker of the M2-like macrophages) and BMP-2 were performed on 4-μm tissue microarray (TMA) sections. Following induction, THP-1 cells were differentiated to M2-like TAMs and were then co-cultured with breast cancer cells (MCF-7). Calcifications and BMP-2 expression were analyzed by Alizarin Red S staining and western blot, respectively.ResultsImmunohistochemical analysis showed that the expression of CD168 was significantly increased in tissues with microcalcifications and was correlated with the expression of BMP-2 and poor prognosis. The formation of cellular microcalcifications and BMP-2 expression were significantly increased in MCF-7 cells co-cultured with TAMs compared with MCF-7 cells alone.ConclusionsThese findings support the hypothesis that TAMs secrete BMP-2 to induce microcalcifications in breast cancer cells and influence prognosis via multiple pathways including BMP-2 and its downstream factors.

Circ0007042 alleviates intervertebral disc degeneration by adsorbing miR-369 to upregulate BMP2 and activate the PI3K/AKt pathway

BackgroundTo identify regulatory ncRNA molecules that can cause differential expression of CDH2 in intervertebral disc degeneration (IDD) and explore whether there are other ways to affect the progression of IDD.MethodsA primary culture of human nucleus pulposus (NP) cells was established and identified by immunofluorescence. An in vitro IDD model was constructed by compressing human NP cells, and the MTT assay was used to measure cell viability. Changes in the ncRNA group were analysed by RNA-seq. The expression levels of hsa_circ_7042, CDH2, and miR-369-3p were detected by qPCR. Cell apoptosis, senescence, and extracellular matrix (ECM) metabolism were detected by flow cytometry, β-galactosidase staining, and Western blotting. hsa_circ_7042, miR-369-3p, and bone morphogenetic protein 2 (BMP2) were verified by luciferase and RNA immunoprecipitation (RIP) analyses. The PI3K/Akt pathway was validated by transfection of BMP2 siRNA. Furthermore, a mouse model of lumbar spine instability was constructed. circ_7042 adenovirus was packaged and injected into the intervertebral discs of mice, and the influence of circ_7042 overexpression on intervertebral disc degeneration was determined.ResultsWestern blotting, qPCR, and flow cytometry analyses confirmed that overexpression of circ_7042 could downregulate miR-369-3p and upregulate the expression of CDH2 and BMP2 in IDD cell and animal models. Additionally, the levels of apoptotic and senescent cells decreased, and ECM degradation decreased. The PI3K/Akt pathway was significantly activated after circ_7042 was overexpressed. The injection of circ_7042-overexpressing adenovirus effectively reduced ECM degradation and the level of apoptosis in NP tissue.Conclusionscirc_7042 could upregulate the expression of CDH2 and BMP2 by absorbing miR-369-3p, and the increased BMP2 activated the PI3K/Akt pathway, thus improving IDD.

Altered Expression of the Hedgehog Pathway Proteins BMP2, BMP4, SHH, and IHH Involved in Knee Cartilage Damage of Patients With Osteoarthritis and Kashin-Beck Disease

ObjectiveTo investigate the expression of Hedgehog (HH) signaling pathway proteins in knee articular cartilage from Kashin-Beck disease (KBD) and osteoarthritis (OA) patients.MethodsKnee articular cartilage samples were collected from normal (N), OA, and KBD adults (aged 38-60 years) and divided into 3 groups with 6 subjects in each group. The localization of the HH pathway proteins bone morphogenetic protein 2 (BMP2), bone morphogenetic protein 4 (BMP4), Sonic hedgehog (SHH), and Indian hedgehog (IHH) was observed with the microscope after immunohistochemical (IHC) staining. Positive staining cell rates of each proteins were compared.ResultsThe strongest stainings of all proteins were observed in the middle zones of all 3 groups. The positive staining rates of BMP4 and IHH were significantly lower in the OA and KBD groups than those in the N group in all 3 zones. The positive staining rates of BMP2 and SHH tend to be lower in the OA and KBD groups than those in the N group in the deep zone, while higher in the OA and KBD groups than those in the N group in superficial and middle zones.ConclusionsAltered expression of the HH pathway proteins BMP2, BMP4, SHH, and IHH was found in OA and KBD articular cartilage. There seemed to be a compensatory effect between SHH and IHH in cartilage damage. Further studies on the pathogenesis of OA and KBD may be carried out from these aspects in the future.

Toll-Like Receptor 4 Gene Knockout Increases the Function of Exogenous Bone Marrow Stem Cells in Ischemic Brain

The present study was to investigate the effects of toll-like receptor 4 on transplanted bone marrow stem cells in the ischemic brain. Focal cerebral ischemia was induced by permanent occlusion of middle cerebral artery in toll-like receptor 4 gene knockout mice and wild type control mice. Bone marrow stem cells were injected into lateral cerebral ventricle. The cylinder tests, in vivo image, polymerase chain reaction array, etc. were applied to evaluate the neurological function and the underlying mechanisms. On the 10th d after permanent occlusion of middle cerebral artery, the ratio of right limb movement in group 1 was significantly higher than that in wild type and toll-like receptor 4 gene knockout mice without injection of bone marrow stem cells (p<0.05). In vivo imaging showed that on the 3rd d after injection of bone marrow stem cells, the fluorescence intensities in group 3 and group 1 were significantly enhanced compared with that on the 1st d and 7th d. There were more green fluorescent protein positive cells in group 3 and group 1 compared with sham controls (p<0.05). Amount of green fluorescent protein positive cells in group 1 was significantly higher compared with group 3. In toll-like receptor 4 gene knockout mice, bone morphogenetic protein 2 messenger ribonucleic acid and fibroblast growth factor 9 messenger ribonucleic acid were up-regulated and interleukin-1-beta protein was decreased, compared with wild type mice. The present study firstly demonstrated that toll-like receptor 4 deficiency further increased the amount of intracerebroventricularly injected bone marrow stem cells in ischemic brain areas, thereby improved neurological function after focal cerebral infarction. The increased expression of bone morphogenetic protein 2 and fibroblast growth factor 9 in the environment of toll-like receptor 4 deficiency might be one of the molecular mechanisms.

Cytotoxicity and bioactive potential of new root repair materials for use with BMP-2 transfected human osteoblast cells.

Modified formulations of calcium silicate repair materials with additives have been developed to enhance handling, consistency, biocompatibility and bioactivity. Considering the relevance of osteoblastic cell response to mineralized tissue repair, human osteoblastic cells (Saos-2 cells overexpressing BMP-2) were exposed to mineral trioxide aggregate (MTA) (with calcium tungstate - CaWO4), MTA HP Repair, Bio-C Repair and Bio-C Pulpo. Cell viability was assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR), and cell death, by flow cytometry. Gene expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX-2), and alkaline phosphatase (ALP) osteogenic markers were evaluated by real-time polymerase chain reaction (RT-qPCR). ALP activity and alizarin red staining (ARS) were used to detect mineralization nodule deposition. Bioactive cements presented no cytotoxic effect, and did not induce apoptosis at the higher dilution (1:12). MTA, Bio-C Repair and Bio-C Pulpo exhibited higher ALP activity than the control group (P < 0.05) after 7 days. MTA, MTA HP and Bio-C Pulpo affected the formation of mineralized nodules (p < 0.05). Exposure to all cement extracts for 1 day increased BMP-2 gene expression. RUNX-2 mRNA was greater in MTA, MTA HP and Bio-C Repair. MTA, MTA HP and Bio-C Pulpo increased the ALP mRNA expression, compared with BMP-2 unexposed cells (P < 0.05). Calcium silicate cements showed osteogenic potential and biocompatibility in Saos-2 cells transfected BMP-2, and increased the mRNA expression of BMP-2, RUNX-2, and ALP osteogenic markers in the BMP-2 transfected system, thereby promoting a cellular response to undertake the mineralized tissue repair.

Polysaccharides from aloe vera target the Wnt/β-catenin pathway to impact the tooth density of pulpitis rats.

To investigate the mechanism of polysaccharides from aloe vera (PAV), a main active ingredient of Aloe vera, treatment in pulpitis rats. Pulpitis were modeled by drilling the occlusal central fossa with Sprague Dawley rats. Next, the rats were treated with 20, 40, and 80 mg/kg PAV for three weeks, respectively. Computed tomography scanning assay, hematoxylin and eosin staining, and tartrate-resistant acid phosphatase staining were used to detect the pathology change. Then, levels of tumor necrosis factor-α, interleukin-1β, prostaglandin E2, and ciclooxigenase 2 were detected by enzyme-linked immunosorbent assay. The expressions of bone morphogenetic protein 2 human (BMP-2), osteocalcin, osterix, and runt-related transcription factor 2 (Runx2) were quantified by quantitative real-time polymerase chain reaction and Western blotting (WB). Finally, Wnt3a expression, p-GSK3β/GSK3β and p-β-catenin/β-catenin ratio were analyzed by WB. PAV up regulated the bone mineral density, and reduced the breakage of the crown and cervical structures, and the necrosis of the crown and root pulp of pulpitis rats. In addition, results indicated that PAV could inhibit osteoblast formation. While osteoblasts' number was decreased, proteins of BMP-2, osteocalcin, osterix, and Runx2 were up-regulated by PAV. Furthermore, PAV increased the Wnt3a expression and the p-β-catenin/β-catenin ratio, and decreased p-GSK3β/GSK3β ratio. Interestingly, these effects were all in dose dependence. PAV could inhibit pulp inflammation and promote osteoblasts differentiation via suppressing the activation of the Wnt/β-catenin signaling, enhancing the dental bone density.

Assessment of BMP responses in an in vitro model of acute ethanol toxicity

The assay presented here was designed to assess the immediate effects of ethanol (EtOH) exposure on intracellular signaling activated by BMPs (Bone Morphogenetic Proteins). Previous reports of the relationship between EtOH exposure and BMP-dependent signaling have primarily assessed the expression of individual BMPs, changes in BMP target genes or effects on the phosphorylation level of key downstream mediators after days or weeks of in vivo EtOH exposure. What happens to BMP-stimulated signaling immediately following exposure to EtOH remains largely unexplored. Here, the early events of BMP-evoked intracellular signaling were examined in an in vitro model of acute EtOH toxicity. The BMP/Ethanol Stimulation Assay involved first stimulating cultured cells with recombinant BMPs. BMP-evoked intracellular signaling was then allowed to develop for 30 minutes. Next, the cells were exposed to a range of EtOH concentrations for an additional 30 minutes. Finally, the cultures were processed for Western blot analysis or immunofluorescent labeling.This short-term assay:• Permits investigation of EtOH exposure during the initial signaling events downstream of BMP receptor activation• Enables assessment of how the presence of BMPs might protect against cellular injury caused by toxic EtOH levels

Overexpression of lncRNA A2M-AS1 inhibits cell growth and aggressiveness via regulating the miR-587/bone morphogenetic protein 3 axis in lung adenocarcinoma.

Lung adenocarcinoma (LUAD) is a malignant tumor that occurs in the lungs. Numerous reports have substantiated the participation of long non-coding RNAs (lncRNAs) in the tumorigenesis of LUAD. Previously, lncRNA alpha-2-macroglobulin antisense RNA 1 (A2M-AS1) was confirmed to be an important regulator in the biological processes of LUAD and dysregulation of A2M-AS1 was associated with non-small cell lung cancer (NSCLC) progression. However, the precise mechanism of A2M-AS1 in LUAD has not been elucidated. Therefore, our study was designed to investigate the detailed molecular mechanism of A2M-AS1 in LUAD. Herein, the expression of lncRNA A2M-AS1, microRNA (miRNA) miR-587, and bone morphogenetic protein 3 (BMP3) in LUAD cell lines and tissues were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting. The viability, proliferation, migration and invasion of LUAD cells were tested by cell counting kit-8 (CCK-8), colony formation and Transwell assays. In vivo tumor growth was investigated by xenograft animal experiment. Interactions among A2M-AS1, miR-587 and BMP3 were measured by RNA pulldown and luciferase reporter assays. In this study, A2M-AS1 was downregulated in LUAD tissues and cells and related to poor prognosis in LUAD patients. A2M-AS1 overexpression suppressed LUAD cell proliferation, migration and invasion in vitro and inhibited tumor growth in vivo. Mechanistically, A2M-AS1 directly bound with miR-587 to promote BMP3 expression in LUAD cells. Low expression of BMP3 was found in LUAD tissues and cells and was closely correlated with poor prognosis in LUAD patients. BMP3 deficiency reserved the inhibitory influence of A2M-AS1 overexpression on LUAD cell behaviors. Overall, A2M-AS1 inhibits cell growth and aggressiveness via regulating the miR-587/BMP3 axis in LUAD.

The Clinical Significance of the Expression of the Serum BMP 6 in the Patients with Tumor-related Anemia with Lower Levels of CRP and Hepcidin

<i>Objective:</i> To research the expression characteristic and clinical significance of the serum bone morphogenetic protein 6 (BMP6) at the lower expression condition of serum C-reactive protein (CRP) and Hepcidin (Hepc) in patients with tumor-related anemia. <i>Methods:</i> 113 hospitalized tumor patients from October 2019 to June 2020 and 20 healthy volunteers (control group) were recruited. We used enzyme-linked immunosorbent assay (ELISA) to determine the levels of BMP6, soluble-hemojuvelin (s-HJV) and Hepc, Electrochemiluminescence method to determine SF level, and particle-enhanced immune transmission turbidity method to determine CRP level, then analyzed their expression characteristics, correlations and clinical significance. <i>Results:</i> (1) The levels of BMP6, Hepc and CRP of anemia group were respectively 426.18±202.83mg/mL, 6.58±5.41ug/L and 19.10±18.24mg/L, which were all higher than 334.37±171.32mg/mL, 4.60±2.28ug/L and 3.62±3.46mg/L of non-anemia group (<i>t</i>=2.5804, 2.4750 and 3.4144, <I>P</I><0.01, <I>P</I><0.05 and <I>P</I><0.01, respectively). (2) The level of s-HJV was 0.71±0.28ng/mL, which was obviously lower than 1.07±1.01ng/mL (<i>t</i>=2.6529, <I>P</I><0.01) in non-anemia group. (3) There were negative correlations between BMP6 and both Hb and s-HJV (respectively <i>r</i>=-0.2807, <i>t</i>=3.0812 and <i>r</i>=-0.2497, <i>t</i>=2.7230, respectively, <I>P</I> both <0.01), but there were no correlations between BMP6 and CRP, Hepc and SF. (4) Hepc was positively correlated with both CRP and SF (<i>r</i>=0.2004, <i>t</i>=2.0660 and <i>r</i>=0.3089, <i>t</i>=3.1045, <I>P</I><0.05 and <I>P</I><0.01, respectively). Hepc was negatively correlated with Hb (<i>r</i>=-0.2524, <i>t</i>=2.7525, <I>P</I><0.01). Hepc had no correlation with s-HJV (<I>P</I>>0.05). (5) The BMP6 in the normal CRP group was 461.86±142.91 mg/mL, which was significantly higher than 364.51± 129.84 mg/mL in elevated CRP group (<I>F</I>=11.35, <I>P</I><0.01). (6) Hepc, s-HJV and SF in the normal CRP group were 5.17±3.49ug/L, 0.69±0.26ng/mL and 149.89±142.34ng/mL, respectively, which were all significantly lower than 9.02±7.14ug/L, 0.76±0.33ng/mL and 288.36±254.51ng/mL in the elevated CRP group (<I>F</I>=9.56, 4.67 and 2.84, <I>P</I>=0.000, 0.09, 0.046, respectively). <i>Conclusions:</i> When tumor patients with anemia is in a low-level condition of CRP and Hepc, the high expression of BMP6 is an important cause for the occurrence and development of anemia.

MGP Regulates Perivascular Adipose-Derived Stem Cells Differentiation Toward Smooth Muscle Cells Via BMP2/SMAD Pathway Enhancing Neointimal Formation

Perivascular adipose-derived stem cells (PV-ADSCs) could differentiate into smooth muscle cells (SMCs), participating in vascular remodeling. However, its underlying mechanism is not well explored. Our previous single-cell RNA-sequencing dataset identified a unique expression of matrix Gla protein (MGP) in PV-ADSCs compared with subcutaneous ADSCs. MGP involves in regulating SMC behaviors in vascular calcification and atherosclerosis. In this study, we investigated MGP’s role in PV-ADSCs differentiation toward SMCs in vitro and in vascular remodeling in vivo. PV-ADSCs were isolated from perivascular regions of mouse aortas. Quantitative reverse transcription–polymerase chain reaction (qRT-PCR), Western blot, and immunofluorescence confirmed higher MGP expression in PV-ADSCs. The MGP secretion increased along PV-ADSCs differentiation toward SMCs in response to transforming growth factor–beta 1 (TGF-β1). Lentivirus knockdown of MGP markedly promoted the bone morphogenetic protein 2 (BMP2) expression and phosphorylation of SMAD1/5/8 in PV-ADSCs, subsequently inhibiting its differentiation toward SMCs. Such inhibition could be partially reversed by further application of BMP2 inhibitors. On the contrary, exogenous MGP inhibited BMP2 expression and SMAD1/5/8 phosphorylation in PV-ADSCs, thereby promoting its differentiation toward SMCs. Transplantation of cultured PV-ADSCs, which was pretreated by MGP knockdown, in mouse femoral artery guide-wire injury model significantly alleviated neointimal hyperplasia. In conclusion, MGP promoted the differentiation of PV-ADSCs toward SMCs through BMP2/SMAD-mediated signaling pathway. This study offers a supplement to the society of perivascular tissues and PV-ADSCs.

Application of Hyaluronic Acid as a Biopolymer Material in Reconstruction of Interdental Papilla in Rats

Applying hyaluronic acid, a biopolymer material, in the treatment of interdental papilla reconstruction has become a trend. The main objective of this research is to investigate the histologic effect of hyaluronic acid on interdental papilla over time. Deficient interdental papilla models were surgically created in sixty-two Sprague-Dawley (SD) rats and were randomly treated with the injection of hyaluronic acid (HA group) or phosphate-buffered saline (sham control group) or left untreated (control group). After 2, 4, and 8 weeks, the rats were sacrificed in batches to observe the histological changes. A fluorochrome label was used to monitor bone formation in 8 weeks. Immunohistochemical analyses were performed to evaluate the expression of potentially relevant cytokines, vascular endothelial growth factor (VEGF), bone morphogenetic protein-2 (BMP-2), and Wnt-induced secreted protein 1 (WISP1) in the gingival tissue in 8 weeks. A preliminary study of HA degradation after 24 weeks was performed in two rats. Following the HA injection, no inflammation or granulomatous foreign body reaction was observed. HA was able to promote collagen fiber and alveolar bone regular formation in the reconstruction site. HA also enhanced VEGF, BMP-2, and WISP-1 expression in gingival tissue (p＜0.05). After 24 weeks, there was no HA filler observed in the interdental papilla. In conclusion, our study suggested that HA is an effective way to reconstruct deficient interdental papilla.

Characterizations and the Mechanism Underlying Osteogenic Activity of Peptides from Enzymatic Hydrolysates of Stichopus japonicus.

Sea cucumber (Stichopus japonicus) is a kind of fishery product with high nutritional value. It exhibits a wide range of biological activity and has potential application in the food, pharmaceutical, and biomedical industries. However, there are no reports available on the effects of S. japonicus peptides (SJP) on bone mineral density regulations. The purpose of this work was to analyze the composition and osteogenic activity of SJP and explore its underlying mechanism. The results showed that SJP stimulated cell proliferation, differentiation, and mineralization in a dose-dependent manner. In addition, SJP could promote the proliferation of MC3T3-E1 cells by altering the cell cycle progression and regulating the expression of Cyclins. Besides, SJP activated the WNT/β-catenin pathway and increased the nuclear level of the active form β-catenin. Furthermore, SJP also induced the expression of bone morphogenetic protein (BMP-2) and increase the phosphorylation levels of p38, JNK, and ERK, suggesting that the osteogenic activity of SJP may be achieved through the activation of WNT/β-catenin and BMP/MAPK signal pathways. In vivo, SJP significantly inhibited the serum levels of RANKL, ALP, and TRAP, whereas it increased the levels of osteocalcin and osteoprotegerin in OVX-mice. These results indicate that SJP may have the potential to stimulate bone formation and regeneration, and may be used as a functional food or nutritional supplement to prevent osteoporosis.

Dysregulated BMP2 in the Placenta May Contribute to Early-Onset Preeclampsia by Regulating Human Trophoblast Expression of Extracellular Matrix and Adhesion Molecules

Many pregnancy disorders, including early-onset preeclampsia (EOPE), are associated with defects in placental trophoblast cell invasion and differentiation during early placental development. Bone morphogenetic protein 2 (BMP2) belongs to the TGF-β superfamily and controls various physiological and developmental processes. However, the expression of BMP2 in the placenta and underlying molecular mechanisms of how BMP2 regulates trophoblast function remain unclear. In this study, we analyzed several publicly available microarray and RNA-seq datasets and revealed differences in expression of TGF-β superfamily members between gestational age-matched non-preeclamptic control and EOPE placentas. Importantly, BMP2 levels were significantly reduced in EOPE placentas compared with controls, and RNAscope in situ hybridization further demonstrated BMP2 expression was disrupted in EOPE placental villi. To explore the molecular mechanisms of BMP2-regulated early trophoblast differentiation, we examined BMP2 expression in first-trimester human placenta and found it to be localized to all subtypes of trophoblasts and the decidua. RNA-seq analysis on control and BMP2-treated primary human trophoblast cells identified 431 differentially expressed genes, including several canonical TGF-β/BMP signaling targets (BAMBI, ID1, INHBA, IGFBP3). Gene ontology annotations revealed that differentially expressed genes were involved in cell adhesion and extracellular matrix organization. Furthermore, we identified adhesion molecule with IgG-like domain 2 (AMIGO2) as a novel target for BMP2 that contributed to BMP2-induced trophoblast invasion and endothelial-like tube formation. Overall, our findings provide insight into the molecular processes controlled by BMP2 during early placental development that may contribute to the pathogenesis of EOPE.

Deficiency of Formyl Peptide Receptor 2 Retards Hair Regeneration by Modulating the Activation of Hair Follicle Stem Cells and Dermal Papilla Cells in Mice.

Hair loss is one of the most common chronic diseases, with a detrimental effect on a patient's psychosocial life. Hair loss results from damage to the hair follicle (HF) and/or hair regeneration cycle. Various damaging factors, such as hereditary, inflammation, and aging, impair hair regeneration by inhibiting the activation of hair follicle stem cells (HFSCs) and dermal papilla cells (DPCs). Formyl peptide receptor 2 (FPR2) regulates the inflammatory response and the activity of various types of stem cells, and has recently been reported to have a protective effect on hair loss. Given that stem cell activity is the driving force for hair regeneration, we hypothesized that FPR2 influences hair regeneration by mediating HFSC activity. To prove this hypothesis, we investigated the role of FPR2 in hair regeneration using Fpr2 knockout (KO) mice. Fpr2 KO mice were found to have excessive hair loss and abnormal HF structures and skin layer construction compared to wild-type (WT) mice. The levels of Sonic hedgehog (Shh) and β-catenin, which promote HF regeneration, were significantly decreased, and the expression of bone morphogenetic protein (Bmp)2/4, an inhibitor of the anagen phase, was significantly increased in Fpr2 KO mice compared to WT mice. The proliferation of HFSCs and DPCs was significantly lower in Fpr2 KO mice than in WT mice. These findings demonstrate that FPR2 impacts signaling molecules that regulate HF regeneration, and is involved in the proliferation of HFSCs and DPCs, exerting a protective effect on hair loss.

Age-associated changes in microRNAs affect the differentiation potential of human mesenchymal stem cells: Novel role of miR-29b-1-5p expression.

Age-associated osteoporosis is widely accepted as involving the disruption of osteogenic stem cell populations and their functioning. Maintenance of the local bone marrow (BM) microenvironment is critical for regulating proliferation and differentiation of the multipotent BM mesenchymal stromal/stem cell (BMSC) population with age. The potential role of microRNAs (miRNAs) in modulating BMSCs and the BM microenvironment has recently gained attention. However, miRNAs expressed in rapidly isolated BMSCs that are naïve to the non-physiologic standard tissue culture conditions and reflect a more accurate in vivo profile have not yet been reported. Here we directly isolated CD271 positive (+) BMSCs within hours from human surgical BM aspirates without culturing and performed microarray analysis to identify the age-associated changes in BMSC miRNA expression. One hundred and two miRNAs showed differential expression with aging. Target prediction and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that the up-regulated miRNAs targeting genes in bone development pathways were considerably enriched. Among the differentially up-regulated miRNAs the novel passenger strand miR-29b-1-5p was abundantly expressed as a mature functional miRNA with aging. This suggests a critical arm-switching mechanism regulates the expression of the miR-29b-1-5p/3p pair shifting the normally degraded arm, miR-29b-1-5p, to be the dominantly expressed miRNA of the pair in aging. The normal guide strand miR-29b-1-3p is known to act as a pro-osteogenic miRNA. On the other hand, overexpression of the passenger strand miR-29b-1-5p in culture-expanded CD271+ BMSCs significantly down-regulated the expression of stromal cell-derived factor 1 (CXCL12)/ C-X-C chemokine receptor type 4 (SDF-1(CXCL12)/CXCR4) axis and other osteogenic genes including bone morphogenetic protein-2 (BMP-2) and runt-related transcription factor 2 (RUNX2). In contrast, blocking of miR-29b-1-5p function using an antagomir inhibitor up-regulated expression of BMP-2 and RUNX2 genes. Functional assays confirmed that miR-29b-1-5p negatively regulates BMSC osteogenesis in vitro. These novel findings provide evidence of a pathogenic anti-osteogenic role for miR-29b-1-5p and other miRNAs in age-related defects in osteogenesis and bone regeneration.

Calcification and Expression of Bone Morphogenetic Protein-4 in Papillary Thyroid Carcinoma

Background/Aim: The purpose of this study was to investigate whether bone morphogenetic protein (BMP)-4 expression differs according to the presence of calcification in papillary thyroid carcinoma and to investigate if calcification and BMP-4 expression are associated with metastasis of papillary carcinoma.Methods: We performed the study using paraffin-embedded tissues of 55 patients who underwent thyroidectomy in Kosin University Gospel Hospital. Of those 55 cases, 36 were papillary thyroid carcinomas, and 19 were benign tumors. Immunohistochemical staining with BMP4 antibody was performed on the paraffin-embedded tissues.Results: Among the 36 papillary thyroid carcinomas, 20 (55.6%) of extrathyroidal invasion and 21 (58.3%) of lymph node metastasis were observed. The presence of calcification was significantly different according to incidence of extranodal invasion (p = 0.02), lymphovascular invasion (p &lt; 0.001), and lymph node metastasis (p = 0.04). BMP-4 expression was weakly positive in benign tumors and strongly positive in papillary thyroid carcinoma (p &lt; 0.001). BMP-4 expression was significantly different by TNM stage (p &lt; 0.001).Conclusion: In the present study, the presence of calcification in thyroid papillary carcinoma was associated with tumor metastasis. BMP-4 was strongly expressed in papillary thyroid carcinoma and was associated with the stage of papillary thyroid carcinoma.

CircSMAD4 alleviates high glucose-induced inflammation, extracellular matrix deposition and apoptosis in mouse glomerulus mesangial cells by relieving miR-377-3p-mediated BMP7 inhibition

BackgroundDiabetic nephropathy (DN) is a common complication of diabetes mellitus. Accumulating studies suggest that the deregulation of circular RNA (circRNA) is involved in DN pathogenesis. This study aimed to investigate the role of circSMAD4 in DN models.MethodsMice were treated with streptozotocin to establish DN models in vivo. Mouse glomerulus mesangial cells (SV40-MES13) were treated with high glucose to establish DN models in vitro. The expression of circSMAD4, miR-377-3p and bone morphogenetic protein 7 (BMP7) mRNA was measured by quantitative real-time PCR (qPCR). The releases of inflammatory factors were examined by ELISA. The protein levels of fibrosis-related markers, apoptosis-related markers and BMP7 were checked by western blot. Cell apoptosis was monitored by flow cytometry assay. The predicted relationship between miR-377-3p and circSMAD4 or BMP7 was validated by dual-luciferase reporter assay or pull-down assay.ResultsCircSMAD4 was poorly expressed in DN mice and HG-treated SV40-MES13 cells. HG induced SV40-MES13 cell inflammation, extracellular matrix (ECM) deposition and apoptosis. CircSMAD4 overexpression alleviated, while circSMAD4 knockdown aggravated HG-induced SV40-MES13 cell injuries. MiR-377-3p was targeted by circSMAD4, and miR-377-3p enrichment partly reversed the effects of circSMAD4 overexpression. BMP7 was a target of miR-377-3p, and circSMAD4 regulated BMP7 expression by targeting miR-377-3p. MiR-377-3p overexpression aggravated HG-induced injuries by suppressing BMP7.ConclusionCircSMAD4 alleviates HG-induced SV40-MES13 cell inflammation, ECM deposition and apoptosis by relieving miR-377-3p-mediated inhibition on BMP7 in DN progression.

Antibacterial wollastonite supported excellent proliferation and osteogenic differentiation of human bone marrow derived mesenchymal stromal cells

Biocompatibility and bacterial infections are the primary concerns associated with the current bone graft substitutes. The application of wollastonite-based scaffolds for bone tissue engineering becomes a novel subject of interest. In the present study, a single phasic wollastonite scaffold was synthesised using citric acid-based sol–gel combustion route. Its physicochemical characteristics, antibacterial properties as well as its biocompatibility and osteogenic induction effect on human bone marrow derived stromal cells (hBMSCs) are yet to be explored. The TGA/DTA, XRD and SEM/EDX confirmed the characteristics of wollastonite. The antibacterial test indicated wollastonite inhibition of 47.81% and 45.54% for gram-positive, Staphylococcus aureus and Staphylococcus epidermidis and 47.04% and 46.07% for gram-negative, Escherichia coli and Pseudomonas aeruginosa bacterial strains, respectively. The SEM micrographs demonstrated an excellent attachment of hBMSCs on wollastonite and comparable to commercial hydroxyapatite (cHA) scaffold. The alamar blue cell proliferation assay confirmed 1.7- and 1.8-fold significant increase in hBMSCs seeded on wollastonite and cHA scaffold, respectively, on day 14 as compared with day 1. The immunohistochemistry analysis on Type-I collagen (Col1) and Bone morphogenetic protein-2 (BMP2) expression on day 14 confirmed the osteogenic differentiation of hBMSCs seeded on wollastonite and comparable with cHA scaffold. In conclusion, wollastonite scaffold has a greater potential to substitute bone grafts in orthopaedic applications.

Designing a novel CaO–MgO–SiO2-based multiphase bioceramic with adjustable ion dissolution behavior for enhancing osteogenesis

Many bioactive inorganic ions such as calcium (Ca), magnesium (Mg) and silicon (Si) have been confirmed to take part in bone regeneration through promoting the secretion of osteogenesis-related growth factors. However, it is not clear about the effective concentration range and action mechanism of ion combinations on synergistically facilitating osteogenesis. Besides, it is of great significance to design a novel bioceramic that could continuously release these effective inorganic ions to match the need for bone tissue repair. Thus, in this work, the concentration level of combined Ca, Mg, Si ions beneficial to osteogenesis of bone marrow-derived mesenchymal stem cells (BMSCs) was explored by orthogonal experimental design (OED) firstly. Meanwhile, eight single-phase ceramics in CaO–MgO–SiO2 system were synthesized, and the influences of their inherent properties on ion dissolution behaviors were systematically investigated. Finally, based on the above results, three multiphase bioceramics (named C1, C2 and C3) containing calcium silicate, diopside and akermanite as crystalline phase were designed and synthesized. It was found that the ionic extracts of bioceramic C1 were just within the above effective concentration range, displaying a predetermined ability of stimulating osteogenic differentiation of BMSCs through enhancing the expression of bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF). All the results indicated that this novel multiphase bioceramic with adjustable ion dissolution behavior has a great application potential in bone tissue engineering.