Journal: Materials Today Bio

Article Title: The combination of a 3D-Printed porous Ti–6Al–4V alloy scaffold and stem cell sheet technology for the construction of biomimetic engineered bone at an ectopic site

doi: 10.1016/j.mtbio.2022.100433

Figure Lengend Snippet: Gene primer sequences for RT‒PCR of BMSCs.

Article Snippet: Primary antibodies, including anti -OCN (Genetex, 1:1000), anti -OPN (Novusbio, 1:1000), anti-COL-I (Bioss, 1:1000), anti -Runx2 (Bioss, 1:1000), and anti -β-actin (Abcam, 1:1000), were added to the membranes.

Techniques:

Journal: Materials Today Bio

Article Title: The combination of a 3D-Printed porous Ti–6Al–4V alloy scaffold and stem cell sheet technology for the construction of biomimetic engineered bone at an ectopic site

doi: 10.1016/j.mtbio.2022.100433

Figure Lengend Snippet: In vitro analysis of osteogenesis of the combination of BMSCs and PT scaffolds. (A) Schematic of the experiment. BMSCs were collected from each group after 7 days and 14 days of coculturing with PT scaffolds in osteogenic induction medium. (B) Alizarin red staining images. (C) Quantification of Alizarin red staining based on the relative absorbance value. (D–G) The RT‒PCR results of osteogenesis-related gene expression of OCN, OPN, COL-I, and Runx2 in the PT, PT/LC, PT/MC, PT/HC, and PT/CS groups on days 7 and 14. These results suggested that increasing the number of initial seeding cells improved the capability of osteogenic differentiation. (H) Western blot (WB) analysis presented results similar to those of RT‒PCR. ∗ indicates significant differences (p ​< ​0.05).

Article Snippet: Primary antibodies, including anti -OCN (Genetex, 1:1000), anti -OPN (Novusbio, 1:1000), anti-COL-I (Bioss, 1:1000), anti -Runx2 (Bioss, 1:1000), and anti -β-actin (Abcam, 1:1000), were added to the membranes.

Techniques: In Vitro, Staining, Gene Expression, Western Blot

Journal: Nanomaterials

Article Title: Antibacterial and Osteogenic Activity of Titania Nanotubes Modified with Electrospray-Deposited Tetracycline Nanoparticles

doi: 10.3390/nano10061093

Figure Lengend Snippet: Osteogenic differentiation of osteoblasts on the specimens for 21 d. ( a ) The expression levels of osteopontin (OPN) and osteocalcin (OCN) assessed by real-time RT-PCR (* p < 0.05 vs. MA). ( b ) Fluorescence images of OCN (upper) and OPN (lower) (scale bar = 100 µm).

Article Snippet: To confirm the findings, immunofluorescence staining was performed with solutions of anti-OPN antibody (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-OCN antibody (QED Bioscience, San Diego, CA, USA) in blocking solution (1% BSA in PBS) at 4 °C for 1 d. After the cells were rinsed with wash buffer, they were soaked in solution with goat anti-mouse lgG-FITC (Santa Cruz Biotechnology, Dallas, TX, USA) for 1 h. The stained cells were observed using CLSM (LSM700, Carl-Zeiss).

Techniques: Expressing, Quantitative RT-PCR, Fluorescence

Journal: Journal of Orthopaedic Surgery and Research

Article Title: T. gondii excretory proteins promote the osteogenic differentiation of human bone mesenchymal stem cells via the BMP/Smad signaling pathway

doi: 10.1186/s13018-024-04839-0

Figure Lengend Snippet: TgEP promoted the osteogenic differentiation of hBMSCs. ( A ) Cytotoxic effect of TgEP on hBMSCs detected using a cytotoxicity detection kit. ( B ) Effect of TgEP on cell proliferation assessed using a CCK-8 kit. ( C ) ALP staining was performed to evaluate ALP expression in the cells. Scale bar = 500 μm. ( D ) Quantification of ALP staining results using ImageJ software and statistical analysis. ( E ) Alizarin red staining was performed to measure the content of mineralized nodules in the cells. Scale bar = 500 μm. ( F ) Quantification of alizarin red staining results using ImageJ software and statistical analysis. ( G ) mRNA transcription levels of osteogenic genes were assessed using qRT‒PCR after the cells were treated for 3 or 7 days. ( H , J ) Western blotting analysis of the osteogenic proteins ALP, Runx2, Osx, and OCN in hBMSCs treated for 5 days ( H ) or 7 days ( J ). The expression levels were quantified using β-actin as a loading control ( I , K ). ( L ) Immunofluorescence staining was performed to detect the expression of Runx2 in the cells. Scale bar = 30 μm. (Data are presented as the mean ± SD; n = 3; * P < 0.05, ** P < 0.01, *** P < 0.001, ns P >0.05 compared to the control group)

Article Snippet: For immunohistochemistry analysis, tissue sections were stained according to standard protocols using anti-osteocalcin (OCN) antibodies (Cat.#: 614,487) (1:100, ZEN-BIO, China).

Techniques: CCK-8 Assay, Staining, Expressing, Software, Western Blot, Control, Immunofluorescence

Journal: Journal of Orthopaedic Surgery and Research

Article Title: T. gondii excretory proteins promote the osteogenic differentiation of human bone mesenchymal stem cells via the BMP/Smad signaling pathway

doi: 10.1186/s13018-024-04839-0

Figure Lengend Snippet: TgEP promotes the repair of bone defects in vivo. ( A ) Histological analysis of bone tissue sections at 6 weeks, including H&E staining, Masson’s trichrome staining, and osteocalcin immunohistochemical staining. Scale bar = 500 μm. ( B ) Histological analysis of bone tissue sections at 8 weeks, including H&E staining, Masson’s trichrome staining, and osteocalcin immunohistochemical staining. Scale bar = 500 μm.

Article Snippet: For immunohistochemistry analysis, tissue sections were stained according to standard protocols using anti-osteocalcin (OCN) antibodies (Cat.#: 614,487) (1:100, ZEN-BIO, China).

Techniques: In Vivo, Staining, Immunohistochemical staining

Journal: Dentistry Journal

Article Title: In Vivo Evaluation of Regenerative Osteogenic Potential Using a Human Demineralized Dentin Matrix for Dental Application

doi: 10.3390/dj12030076

Figure Lengend Snippet: Representative histological images of (H&E) ( A , D , G , J ), Masson’s trichrome ( B , E , H , K ) and osteocalcin immune staining ( C , F , I , L ) are shown at 4 weeks post-treatment. Coronal sections through the mid-point of the defects were prepared from decalcified specimens. Fibrous connective tissues were seen to be mostly filled in the defect areas in the control group ( A – C ). The group treated with SA hydrogel as the only grafted material showing fibrous and adipose tissue ( D – F ). lacuna spaces were observed in the newly formed lamellar bone in NHH with a wide bone marrow space ( G – I ). DDMH-treated groups showed considerable new bone trabeculae formation ( J – L ). BT: bone trabeculae; FT: fibrous tissue. Arrows represent MT- and OCN-positive reactions. Scale bar: 100 μm.

Article Snippet: Subsequently, the samples were embedded in paraffin; and after staining with hematoxylin and eosin (H&E), Masson’s trichrome (MT) (Cat# HT15, Sigma Aldrich), and immunohistochemical staining with a polyclonal anti-rabbit osteocalcin antibody (OCN; Cat# GB11233, diagnostic biosystems) at a 1:150 dilution, histological examination was conducted using an Olympus BX-51 optical microscope (Olympus Co., Tokyo, Japan).

Techniques: Staining, Control

Journal: Dentistry Journal

Article Title: In Vivo Evaluation of Regenerative Osteogenic Potential Using a Human Demineralized Dentin Matrix for Dental Application

doi: 10.3390/dj12030076

Figure Lengend Snippet: Representative histological images of (H&E) ( A , D , G , J ), Masson’s trichrome ( B , E , H , K ) and osteocalcin immune staining ( C , F , I , L ) are shown at 8 weeks post-treatment. Rather than the newly created bone, the defect locations in the control group were found to be primarily filled with fibrous connective tissues and adipose tissue ( A – C ). Circular new bony islands with the presence of early woven bone were observed in the group treated with SA as the only grafted material ( D – F ). Newly formed lamellar bone was observed in NHH with a wide bone marrow space ( G – I ). In contrast, DDMH-treated groups showed considerable new bone formation at 8 weeks with relatively narrow marrow spaces ( J – L ). BT means bone trabeculae; FT: fibrous tissue. Arrows represent MT- and OCN-positive reactions. Scale bar: 100 μm.

Article Snippet: Subsequently, the samples were embedded in paraffin; and after staining with hematoxylin and eosin (H&E), Masson’s trichrome (MT) (Cat# HT15, Sigma Aldrich), and immunohistochemical staining with a polyclonal anti-rabbit osteocalcin antibody (OCN; Cat# GB11233, diagnostic biosystems) at a 1:150 dilution, histological examination was conducted using an Olympus BX-51 optical microscope (Olympus Co., Tokyo, Japan).

Techniques: Staining, Control

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Osteoprogenitors in the bone marrow. The ratio of CD34 + OCN + cells from MDS patients was less than that of normal controls (*P<0.05), and those from Group 2 were great less than that of normal controls (**P<0.01). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; PE, phycoerythrin; PerCP, peridinin chlorophyll.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences) and anti-OCN-PE (cat. no. 564146, BD Biosciences) staining was used to identify osteoprogenitor cells in the bone marrow.

Techniques: Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Activity of osteoblasts. The percentage of OCN + CD34 − Lin − cells among osteoblasts from MDS patients was less than that of normal controls (*P<0.05). The percentage in Group 1 was close to that of normal controls (P>0.05), whereas the percentage in Group 2 was significantly less than that of normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; APC, allophycocyanin; PerCP, peridinin chlorophyll.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences) and anti-OCN-PE (cat. no. 564146, BD Biosciences) staining was used to identify osteoprogenitor cells in the bone marrow.

Techniques: Activity Assay, Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Correlation between the activity of osteoblasts from MDS patients and the severity of MDS. The percentage of OCN + CD34 − Lin − cells among osteoblasts from MDS patients was negatively correlated with the blast count in bone marrow (P<0.01) and was positively correlated with hemoglobin level (P<0.05) and neutrophil count (P<0.05) in the peripheral blood of the MDS patients. MDS, myelodysplastic syndrome; OCN, osteocalcin.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences) and anti-OCN-PE (cat. no. 564146, BD Biosciences) staining was used to identify osteoprogenitor cells in the bone marrow.

Techniques: Activity Assay

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: JAG1 expression on osteoblasts. The percentage of JAG1 + cells out of the total number of OCN + CD34 − Lin − osteoblasts was higher in MDS patients than in normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; JAG1, Jagged 1.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences) and anti-OCN-PE (cat. no. 564146, BD Biosciences) staining was used to identify osteoprogenitor cells in the bone marrow.

Techniques: Expressing, Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: TIM3 expression on osteoblasts. The percentage of TIM3 + CD34 + Lin − cells among osteoblasts was higher in MDS patients than in normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; TIM3, T-cell immunoglobulin and mucin domain-containing 3.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences) and anti-OCN-PE (cat. no. 564146, BD Biosciences) staining was used to identify osteoprogenitor cells in the bone marrow.

Techniques: Expressing, Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Osteoprogenitors in the bone marrow. The ratio of CD34 + OCN + cells from MDS patients was less than that of normal controls (*P<0.05), and those from Group 2 were great less than that of normal controls (**P<0.01). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; PE, phycoerythrin; PerCP, peridinin chlorophyll.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences), anti-Lin-FITC (cat. no. 562722 BD Biosciences) and anti-OCN-allophycocyanin (APC) (cat. no. 557833 BD Biosciences) staining was used to measure the normal osteogenic function by osteoblasts cultured in vitro .

Techniques: Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Activity of osteoblasts. The percentage of OCN + CD34 − Lin − cells among osteoblasts from MDS patients was less than that of normal controls (*P<0.05). The percentage in Group 1 was close to that of normal controls (P>0.05), whereas the percentage in Group 2 was significantly less than that of normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; APC, allophycocyanin; PerCP, peridinin chlorophyll.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences), anti-Lin-FITC (cat. no. 562722 BD Biosciences) and anti-OCN-allophycocyanin (APC) (cat. no. 557833 BD Biosciences) staining was used to measure the normal osteogenic function by osteoblasts cultured in vitro .

Techniques: Activity Assay, Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: Correlation between the activity of osteoblasts from MDS patients and the severity of MDS. The percentage of OCN + CD34 − Lin − cells among osteoblasts from MDS patients was negatively correlated with the blast count in bone marrow (P<0.01) and was positively correlated with hemoglobin level (P<0.05) and neutrophil count (P<0.05) in the peripheral blood of the MDS patients. MDS, myelodysplastic syndrome; OCN, osteocalcin.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences), anti-Lin-FITC (cat. no. 562722 BD Biosciences) and anti-OCN-allophycocyanin (APC) (cat. no. 557833 BD Biosciences) staining was used to measure the normal osteogenic function by osteoblasts cultured in vitro .

Techniques: Activity Assay

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: JAG1 expression on osteoblasts. The percentage of JAG1 + cells out of the total number of OCN + CD34 − Lin − osteoblasts was higher in MDS patients than in normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; JAG1, Jagged 1.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences), anti-Lin-FITC (cat. no. 562722 BD Biosciences) and anti-OCN-allophycocyanin (APC) (cat. no. 557833 BD Biosciences) staining was used to measure the normal osteogenic function by osteoblasts cultured in vitro .

Techniques: Expressing, Standard Deviation

Journal: Oncology Letters

Article Title: Abnormal changes in the quantity and function of osteoblasts cultured in vitro in patients with myelodysplastic syndrome

doi: 10.3892/ol.2018.9206

Figure Lengend Snippet: TIM3 expression on osteoblasts. The percentage of TIM3 + CD34 + Lin − cells among osteoblasts was higher in MDS patients than in normal controls (*P<0.05). Data are presented as the mean ± standard deviation. MDS, myelodysplastic syndrome; OCN, osteocalcin; TIM3, T-cell immunoglobulin and mucin domain-containing 3.

Article Snippet: Anti-CD34-PerCP (cat. no. 555823 BD Biosciences), anti-Lin-FITC (cat. no. 562722 BD Biosciences) and anti-OCN-allophycocyanin (APC) (cat. no. 557833 BD Biosciences) staining was used to measure the normal osteogenic function by osteoblasts cultured in vitro .

Techniques: Expressing, Standard Deviation