Experimental Study on the Construction of Tissue Engineered Bone by Bone Marrow Mesenchymal Stem Cells Induced by MiR-29b Composite Bionic Scaffold

2019 ◽  
Vol 9 (12) ◽  
pp. 1763-1769
Author(s):  
Peng Yu ◽  
Jun Li ◽  
Bingshen Jia ◽  
Zizhenbiao Wang ◽  
Sheng Wang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Type I Collagen ◽  
Negative Control ◽  
Control Group ◽  
Type I ◽  
Mineral Density ◽  
Alp Activity ◽  
Bionic Scaffold ◽  
Marrow Mesenchymal Stem Cells

MiR-29b promotes osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Our study aims to evaluate MiR-29b's role in composite bionic scaffold-induced BMSCs in the construction of tissue engineered bone. Rat BMSCs were isolated and transfected with NC (negative control) and MiR-29b plasmid. Cell proliferation was assessed by MTT assay and the expression of osteogenic genes Runx2 and OC was analyzed by Real time PCR. Healthy male SD rats were divided into fracture group; negative control group; and MiR-29b group followed by analysis of the changes of bone mineral density, ALP activity, and expression of MiR-29b, type I collagen and Runx2 by Real time PCR. Up-regulation of MiR-29b significantly promoted BMSCs cell proliferation, inhibited Caspase 3 activity, and promoted Runx2 and OC expression compared to NC group (P < 0 05). NC group showed significantly increased bone density, ALP activity and the expression of type I collagen and Runx2 compared with control group (P < 0 05). Transfection of BMSCs induced by MiR-29b combined with biomimetic scaffold significantly promoted the expression of MiR-29b in fractured rats, increased bone mineral density and ALP activity, as well as upregulated type I collagen and Runx2, compared to NC group (P < 0 05). Up-regulation of MiR-29b promotes cell proliferation and osteogenic differentiation of BMSCs. Implantation of BMSCs induced by MiR-29b composite bionic scaffold can promote osteogenic differentiation and promote bone healing in bone defects.

Effect of Phosphatase and Tensin Homolog 12 on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells During Aging by Regulating Transforming Growth Factor Beta/Smad Signaling Pathway

2021 ◽  
Vol 11 (8) ◽  
pp. 1630-1635
Author(s):  
Bin Wu ◽  
Fenghua Bai ◽  
Jianping Lin ◽  
Guangji Wang ◽  
Wentao Cai ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Type I Collagen ◽  
Type I ◽  
Smad Signaling ◽  
Alp Activity ◽  
Smad Signaling Pathway ◽  
Marrow Mesenchymal Stem Cells

Aging affects bone marrow mesenchymal stem cells (BMSC) differentiation. PTEN12 regulates cell proliferation and apoptosis. However, whether PTEN12 affects BMSCs osteogenic differentiation during aging is unknown. Two BMSCs derived from Zempster24−/− (senescence) and Zempster24+/+ (normal) mice were cultured in vitro. Real-time PCR analysis was used to analyze PTEN12 expression. PTEN12 siRNA was transfected into senescent Zempster24-/-BMSCs and after 14 days of osteogenic induction, cell proliferation was analyzed by MTT method along with measuring expression of osteocalcin, type I collagen, RUNX2 and OPN by Real time PCR, ALP activity, and TGFβ/smad signaling protein expression by Western blot. Compared to normal BMSCs, PTEN12 level in aging BMSCs was significantly elevated, osteocalcin, type I collagen, RUNX2 and OPN mRNA level was decreased along with reduced ALP activity and TGFβ1 and Smad2 expression (P < 0.05). PTEN12 siRNA transfection into senescent BMSCs significantly down-regulated PTEN12, upregulated osteocalcin, type I collagen, RUNX2 and OPN mRNA, increased ALP activity and TGFβ1 and Smad2 expression (P <0.05). Aging increases PTEN12 level and inhibits BMSCs osteogenic differentiation. Down-regulation of PTEN12 in BMSCs during aging can promote BMSCs osteogenic differentiation by regulating TGFβ/smad signaling pathway.

Progranulin Promotes Osteogenic Differentiation and Osteosynthesis of Bone Marrow Mesenchymal Stem Cells and Alleviates Osteoporosis Through Phosphatidylinositol 3-Kinase/Protein Kinase B/Peroxisome Proliferator-Activated Receptor γ Pathway

2020 ◽  
Vol 10 (12) ◽  
pp. 1865-1870
Author(s):  
Yang Ying ◽  
Binghao Zhao ◽  
Wei Qian ◽  
Li Xu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Mrna Level ◽  
Control Group ◽  
Peroxisome Proliferator ◽  
Mineral Density ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) have self-renewal potential with multi-directional differentiation. Progranulin prevents bone degradation, inhibits inflammation and protects bone tissue. However, the role of Progranulin in osteoporotic BMSCs is unclear. Osteoporosis (OP) rat models were prepared by ovarian removal and treated with different doses (5 and 10 μM) of Progranulin followed by analysis of BMP-2 level by ELISA, bone mineral density and ALP activity. OP rat BMSCs were isolated and assigned into control group and Progranulin group followed by analysis of Progranulin level by ELISA, cell proliferation by MTT assay, RUNX2 and COL1A1 mRNA level by Real time PCR, and PI3K/Akt/PPARγ signaling protein level by Western blot. Progranulin treatment of OP rats dose-dependently increased BMP-2 expression, bone density and ALP activity. Compared with OP group, there were significant differences (P <0.05). Progranulin expression and BMSCs proliferation was increased, and RUNX2 and COL1A1 mRNA expression was elevated in Progranulin-treated OP group along with increased PI3K/Akt expression and decreased PPARγ protein expression. Compared with OP group, the difference was statistically significant, and the change was more significant with increasing concentration (P <0.05). Progranulin promotes BMSCs osteogenic differentiation and proliferation by regulating PI3K/Akt/PPARγ signaling pathway, which is beneficial for OP rats’ bone synthesis.

Effect of Silent Information Regulator on the Survival and Osteogenic Differentiation of Inflammatory Bone Marrow Mesenchymal Stem Cells by Regulating NF-κB Signaling Pathway

2020 ◽  
Vol 10 (1) ◽  
pp. 121-126
Author(s):  
Wenkun Lu ◽  
Tao Wang ◽  
Xunjian Gao ◽  
Fuqiang Yang ◽  
Jianjian Ge
Keyword(s):  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Caspase 3 ◽  
Adipogenic Differentiation ◽  
Control Group ◽  
Promote Cell Proliferation ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells ◽  
Inflammation Group

Osteogenic differentiation of BMSCs is beneficial for osteoarthritis (OA) treatment. Silent information regulator (SIRT1) plays a role in endocrine diseases and aging-related diseases. However, the role of SIRT1 in OA has not yet been elucidated. Rat BMSCs were isolated and divided into control group, inflammation group (BMSCs were cultured with IL-6), SIRT1 group (SIRT1 agonist Resveratrol was added under the action of IL-6) followed by analysis of cell proliferation by MTT assay, Caspase 3 activity, ALP activity, expression of osteogenic genes Runx2 and OC and adipogenic differentiation gene PPARγ2 by Real time PCR, NF-κB expression by western blot and secretion of TNF-α and IL-6 by ELISA. In inflammation group, SIRT1 expression was significantly decreased, cell proliferation was significantly inhibited, and Caspase 3 activity was increased. Meanwhile, ALP activity, Runx2 and OC expression was decreased, PPARγ2 and NF-κB expression was increased, along with elevated TNF-α and IL-6 secretion compared to control (P < 0.05). Resveratrol can significantly promote the expression of SIRT1 in BMSCs of inflammation group, promote cell proliferation, decrease Caspase 3 activity, and increase Runx2 and OC expression. In addition, it decreased PPARγ2 and NF-κB expression and reduced the secretion of TNF-α and IL-6 (P < 0.05). The expression of SIRT1 was decreased in BMSCs under inflammation. SIRT1 overexpression in BMSCs under inflammation inhibits inflammation, promotes proliferation and osteogenic differentiation of BMSCs through regulating NF-κB signaling pathway.

Effect of Hepatic Kinase B1 (LKB1) on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells During Senescence

2020 ◽  
Vol 10 (2) ◽  
pp. 246-251
Author(s):  
Wenxiao Jiang ◽  
Yijun Zhang ◽  
Ye Huang ◽  
Yunfeng Cheng ◽  
Zhigang Liu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Type I Collagen ◽  
Mtor Pathway ◽  
Nodule Formation ◽  
Type I ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Hepatic kinase B1 (LKB1) is a tumor suppressor and regulates cell proliferation and apoptosis. However, whether LKB1 affects bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation of during aging remains unclear. Two BMSCs derived from Zempster24−/− (aging) and Zempster24+/+ (normal) mice were cultured in vitro followed by measurement of LKB1 expression by real-time quantitative PCR and Western blot. LKB1 siRNA was transfected into Zempster24−/−BMSCs and LKB1 expression was measured. 14 days after osteogenic induction, mineralized nodule formation was evaluated by alizarin red staining, expression of Calcin, type I collagen, RUNX2 and OPN mRNA expression was measured, together with alkaline phosphatase (ALP) activity and the PI3K/mTOR pathway activity. Compared with normal BMSCs, LKB1 expression was significantly increased, calcified nodules were decreased, with reduced expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA as well as decreased ALP activity and PI3K/mTOR signaling protein expression (P < 0.05). LKB1 siRNA transfection into senescent BMSCs down-regulated LKB1 expression, increased calcification nodule formation, expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA, as well as increased ALP activity and PI3K/mTOR pathway protein expression (P < 0.05). Aging can promote the increase of LKB1 expression and inhibit the osteogenic differentiation of BMSCs. Down-regulation of LKB1 expression in BMSCs during senescence can promote osteogenic differentiation through regulating PI3K/mTOR pathway.

Effect of Chordin-Like 1 on Differentiation of Bone Marrow Mesenchymal Stem Cells Through Wnt5/TGF-β Signaling Pathway

2019 ◽  
Vol 9 (9) ◽  
pp. 1304-1310
Author(s):  
Qing Yang ◽  
Lei Wu ◽  
Yang Liu ◽  
Bing Yuan
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Control Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Chordin-like 1 (CHRDL1) functions in multiple tissues and organs. However, whether CHRDL1 affects bone marrow mesenchymal stem cells (BMSCs) differentiation remain unclear. Rat BMSCs were isolated and divided into control group, CHRDL1 group and CHRDL1 siRNA group followed by analysis of CHRDL1 level by real time PCR and ELISA, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, expression of o Runx2, OC and PPARγ2 by Real time PCR, TGF-β secretion by ELIS, and Wnt5 protein expression by Western blot. CHRDL1 expression was significantly increased in CHRDL1 group, along with significantly promoted cell proliferation, decreased Caspase 3 activity, increased ALP activity and expression of Runx2 and OC, decreased PPARγ2 expression, increased TGF-β secretion and Wnt5 expression compared to control group (P < 0.05). However, CHRDL1 siRNA transfection significantly decreased CHRDL1 expression, inhibited cell proliferation, increased Caspase 3 activity, decreased ALP activity and Runx2 and OC expression, increased PPARγ2 expression, decreased TGF-β secretion and Wnt5 expression. (P < 0.05). Down-regulation of CHRDL1 expression in BMSCs promotes Wnt5/TGF-β signaling transduction, which in turn increases BMSCs proliferation and osteogenic differentiation. Up-regulation of CHRDL1 expression in BMSCs inhibited the activation of Wnt5/TGF-β signaling pathway, promoted BMSCs apoptosis, and inhibited BMSCs proliferation and osteogenic differentiation.

Distal Low Homeobox 3 Promotes Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation and Bone Synthesis and Ameliorates Osteoporosis by Activating Typical Wnt Signaling

2019 ◽  
Vol 9 (12) ◽  
pp. 1776-1782
Author(s):  
Yongyi Xu ◽  
Lei Chen
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Density ◽  
Osteogenic Differentiation ◽  
Caspase 3 ◽  
Type I Collagen ◽  
Type I ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

The distal low homeobox 3 (DLX3) regulates the bone marrow mesenchymal stem cells (BMSC) osteogenic differentiation. However, whether DLX3 affects osteoporosis (OP) remains unclear. An OVX-induced OP rat model was constructed and DLX3 plasmid was injected followed by analysis of bone mineral density and ALP activity. Rat BMSCs were isolated and divided into control group, OP group and DLX3 group (transfected with DLX3 plasmid) followed by analysis of chondrocytes survival rate by MTT assay, Caspase 3 activity, type I collagen and Osterix expression by Real time PCR and -catenin level by Western blot. DLX3 expression was significantly down-regulated in OP rats with deceased bone density and ALP activity compared to sham group (P < 0 05). When DLX3 was transfected into OP rats, DLX3 expression was significantly up-regulated with increased bone density and ALP activity compared with OP group (P < 0 05). BMSCs survival was significantly decreased in OP group and Caspase 3 activity was significantly increased with downregulated type I collagen, Osterix and -catenin (P < 0 05). However, transfection of DLX3 plasmid into OP group BMSCs cells can significantly reverse the above changes, compared to OP group (P < 0 05). DLX3 expression is reduced in osteoporosis. Up-regulation of DLX3 can promote osteogenic differentiation of BMSCs by regulating typical Wnt signaling, promote differentiation into osteoblasts, increase bone density increase, and then ameliorate osteoporosis.

(deleted)

2021 ◽  
Author(s):  
MV Osikov ◽  
EV Davydova ◽  
KS Abramov
Keyword(s):  
Bone Healing ◽  
Standard Therapy ◽  
Type I Collagen ◽  
Femoral Shaft ◽  
Control Group ◽  
Blood Levels ◽  
Type I ◽  
Mineral Density ◽  
Fracture Consolidation ◽  
To Receive

Efferent physical therapy holds promise as an adjunct to the combination treatment of femoral fractures in young, working-age individuals. The aim of the study was to investigate the dynamics of bone turnover markers at different stages of femoral fracture consolidation in patients undergoing ozone therapy. The study enrolled 20 men (group 2, 47.8 ± 3.5 years) with a femoral shaft fracture (AO/ASIF 32А, 32В). The control group (group 1, 46.8 ± 3.7 years) comprised 10 healthy males. Subgroup 2a (n = 10) was assigned to receive standard therapy; subgroup 2b (n = 10) was assigned to receive standard therapy complemented by minor autohemotherapy (MAHT) at 20 mg/L ozone concentrations. On days 7, 30 and 90, fracture consolidation was assessed on the RUST scale and blood levels of С-terminal telopeptides of type I collagen (bCTx, pg/ml) and procollagen type I carboxy-terminal propeptide (PICP, ng/ml) were measured. On day 7, the total RUST score in subgroups 2a and 2b was 4 points; on day 30, it was 6.5 and 8.7 points, respectively, and on day 90, it reached 10 and 11.5 points, respectively. Bone mineral density was as high as 90% in the MAHT subgroup vs. 78% in subgroup 2а, indicating faster bone healing. On day 30, bCTx levels in subgroup 2b were higher than in subgroup 2a (2289.4 [2145.3; 2365.4] vs. 1894.6 [1745.3; 2098.2], respectively. On day 7, PICP was significantly elevated in subgroup 2b in comparison with subgroup 2a; its levels peaked on days 30 and 90 (day 30: 268.3 [231.2; 286.3] vs. 183.2 [174.6; 195.6]; day 90: 584.6 [512.3; 589.3] vs. 351.2 [312.3; 369.4]. Thus, MAHT produces a positive effect on the quality and intensity of bone healing in men with isolated closed femoral shaft fractures.

Cellular Compatibility of a New Tantalum-Niobium Scaffold Material

2020 ◽  
Author(s):  
Jianan Ouyang ◽  
Zhenhan Deng ◽  
Kang Chen ◽  
Jianyi Xiong ◽  
Ying Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Type I Collagen ◽  
Material Surface ◽  
Control Group ◽  
Type I ◽  
Scaffold Material ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Porous Tantalum ◽  
Significant Difference

Abstract [Objective] To determine the cellular compatibility of porous tantalum-niobium (Ta-Nb) material. [Method] Rabbit osteoblasts were co-cultured with porous Ta-Nb material. The cell proliferation was detected by CCK-8 method, and the cell adhesion was observed under scanning electron microscope (SEM). The expressions of type-I collagen and osteocalcin were detected by RT-PCR assay. [Results] CCK-8 detection indicated that the cell proliferation on the porous Ta-Nb material showed no difference from that of the control group (P>0.05). SEM revealed that a large amount of cells adhered onto the surface and in the pores of the material. The number of cells on the material surface increased obviously over time. RT-PCR assay showed that with the prolonging of the time of co-culture, the expression of type-I collagen was enhanced (P<0.05), while the osteocalcin expression exhibited no significant difference (P>0.05[Conclusion] Porous Ta-Nb scaffold material can be used to promote the adhesion, growth and differentiation of osteoblasts with satisfactory cellular compatibility.

Effect of αCGRP on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Through Smad/Runx2 Signaling Pathway

2020 ◽  
Vol 10 (6) ◽  
pp. 868-873
Author(s):  
Shengxiang Huang ◽  
Haibo Mei ◽  
Rongguo He ◽  
Kun Liu ◽  
Jin Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

The α-calcitonin gene-related peptide (α-CGRP) regulates bone metabolism and has potential applications in enhancing bone remodeling in vivo. However, α-CGRP's role in bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation remain unclear. Rat BMSCs were separated into control group, α-CGRP group and α-CGRP siRNA group, in which BMSCs were transfected with α-CGRP plasmid and α-CGRP siRNA respectively followed by analysis of α-CGRP level by real time PCR and ELISA, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of calcified nodules by alizarin red staining, Smad1 and Smad7 level by Western blot and Runx2 by real time PCR. αCGRP transfection into BMSCs significantly up-regulated CGRP, which could promote cell proliferation, inhibit Caspase 3 activity, promote ALP activity, increase calcified nodules formation and upregulate Smad1, Smad7 and Runx2 compared to control (P < 0.05); transfection of αCGRP siRNA significantly down-regulated CGRP in BMSCs, inhibited cell proliferation, promoted Caspase 3 activity, inhibited ALP activity, inhibited calcified nodules formation and downregulate Smad1, Smad7 and Runx2 (P < 0.05). αCGRP overexpression promotes the Smad/Runx2 signaling, which in turn promotes BMSCs proliferation and osteogenesis. Decreased αCGRP level inhibits Smad/Runx2 signaling, promotes BMSCs apoptosis, inhibits proliferation and osteogenic differentiation.

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