normal human fetal osteoblast cell line  (ATCC)

Journal: Biomedicines

Article Title: CAPE Derivatives as Potent Agents for Induction of Osteogenic Differentiation in DPSCs and Biomaterial Development

doi: 10.3390/biomedicines13123039

Figure Lengend Snippet: Cytotoxicity assessment of biomaterials enriched with CAPE derivatives on hFOB 1.19 osteoblasts. ( a ) MTT assay conducted by using 24 h extracts of the biomaterials after 48 h cell culture (PS medium—polystyrene extract served as negative control of cytotoxicity; * statistically significant results compared to PS medium, # statistically significant results compared to mat_control, $ statistically significant results compared to mat_ 1d _L, ^ statistically significant results compared to mat_ 1a _L, p < 0.05, One-way ANOVA followed by Tukey’s test); ( b ) confocal laser scanning microscope images showing live/dead staining of cells cultured on the surface of biomaterials for 48 h (Nomarski contrast was applied to show biomaterial microstructure; viable cells—green fluorescence; dead cells—red fluorescence; magnified 100×, scale bar = 200 µm).

Article Snippet: Normal human fetal osteoblast cell line (hFOB 1.19, ATCC CRL-11372) was cultured in a 1:1 mixture of Dulbecco’s Modified Eagle’s Medium and Ham’s F12 Medium, supplemented with 2.5 mM L-glutamine, 0.3 mg/mL G418 (Sigma-Aldrich Chemicals, Warsaw, Poland), and 10% FBS (Pan-Biotech GmbH, Aidenbach, Bavaria, Germany) and maintained at 34 °C (5% CO 2 in air atmosphere).

Techniques: MTT Assay, Cell Culture, Negative Control, Control, Laser-Scanning Microscopy, Staining, Fluorescence

Journal: Biochemistry and Biophysics Reports

Article Title: Development of injectable cuttlebone derived nanohydroxyapatite hydrogel for osteoblast cell encapsulation

doi: 10.1016/j.bbrep.2025.102268

Figure Lengend Snippet: Demonstrating characteristics of calcium carbonate microcapsules (CaCO 3 ) without (CaCO 3 alone) and with quercetin (CaCO 3 -QT), (A) Scanning electron microscopic images exhibiting size, shape and surface microstructure of the CaCO 3 -alone (a) and CaCO 3 -QT (b), (B) A cumulative release profile of quercetin (QT) from the CaCO 3 -QT in phosphate buffer saline (PBS), (D) Percentages of cell viability and (E) Percentages of in vitro mineralization derived from alizarin red staining of human fetal osteoblast cell line (hFOB) cultured in growth mediums (D) and osteogenic medium (E) that were pre-incubated with the 2 % (w/v) CaCO 3 alone and CaCO 3 -QT. Abbreviation Control represents the regular growth mediums containing 10 % fetal bovine serum alone and Control-QT, for growth medium supplemented with 10 μg/ml QT in (C and D). The percentages of cell viability and mineralization were calculated relative to a control group. Symbols A – F indicate statistically different among investigation times at p < 0.05, and ∗ represent the differences at p < 0.05, ∗∗p < 0.01 and ∗∗∗∗p < 0.0001 (n = 5, Mean ± SD).

Article Snippet: The human fetal osteoblast cell line (hFOB 1.19) (Cat No. CRL-11372, ATCC, USA) was cultured in a growth medium for cell expansion and an osteogenic medium to stimulate osteogenic differentiation.

Techniques: Calcium Carbonate, Saline, In Vitro, Derivative Assay, Staining, Cell Culture, Incubation, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Development of injectable cuttlebone derived nanohydroxyapatite hydrogel for osteoblast cell encapsulation

doi: 10.1016/j.bbrep.2025.102268

Figure Lengend Snippet: Illustrating the effectiveness of a 5 % (w/v) cuttlebone derived nanohydroxyapatite (CB-nHA) combined with 1 % (w/v) calcium carbonate microcapsules containing quercetin (CaCO 3 -QT) within a thermosensitive hydrogel composed of a 7:1 (w/w) ratio of chitosan to collagen. The goal is to promote cell growth and osteogenic differentiation of the human fetal osteoblast cell line (hFOB). Figures (A–D) show live/dead cell staining of hFOB in a growth medium: (A) on a culture plate and (B–D) encapsulated in hydrogels. Specifically, (B) illustrates hFOB with 0 % CB-nHA-CaCO 3 alone, (C) shows 5 % CB-nHA-CaCO 3 alone, and (D) displays 5 % CB-nHA-CaCO 3 -QT. Figure (E) presents the cell viability percentages of hFOB cultured in mediums pre-incubated with hydrogels for 1, 3, and 5 days, compared to a control group in regular growth medium (Control). Figure (F) depicts alkaline phosphatase (ALP) activity levels of encapsulated cells in the hydrogels containing 0 % and 5 % CB-nHA with either CaCO 3 alone or with CaCO 3 -QT. Statistical significance is indicated as follows: ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, ∗∗∗∗ for p < 0.0001, and ns for p > 0.05 (n = 5, Mean ± SD).

Article Snippet: The human fetal osteoblast cell line (hFOB 1.19) (Cat No. CRL-11372, ATCC, USA) was cultured in a growth medium for cell expansion and an osteogenic medium to stimulate osteogenic differentiation.

Techniques: Derivative Assay, Calcium Carbonate, Staining, Cell Culture, Incubation, Control, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: A ‘Spicy’ Mechanotransduction Switch: Capsaicin-Activated TRPV1 Receptor Modulates Osteosarcoma Cell Behavior and Drug Sensitivity

doi: 10.3390/ijms26188816

Figure Lengend Snippet: 1 Hz–24 h mechanical stretch induces cell orientation in U-2 OS cells via TRPV1 activation. ( A ) Representative fluorescence microscopy images of U-2 OS cells stained with Hoechst (nuclei), β-tubulin (cytoskeleton), and merged channels. Images were acquired at 40× magnification. Scale bar = 25 μm. ( B ) Schematic representation of nuclear orientation expressed as ⟨cos 2 φ⟩. ( C ) Violin plots display the distribution of nuclear orientation values in SAOS-2, U-2 OS, and hFOB cells. ( D ) Violin plots show nuclear orientation in stretched U-2 OS cells treated with AMG (I), Cytochalasin D (Cyto D) or Indomethacin (Indo). All analyses were performed on three biological replicates, with at least 45 cells analyzed per condition.

Article Snippet: Osteosarcoma cell lines Human SAOS-2 (HTB-85) and U-2 OS (HTB-96) and human fetal osteoblast cell line hFOB (CRL-3602) were purchased from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Activation Assay, Fluorescence, Microscopy, Staining

Journal: International Journal of Molecular Sciences

Article Title: A ‘Spicy’ Mechanotransduction Switch: Capsaicin-Activated TRPV1 Receptor Modulates Osteosarcoma Cell Behavior and Drug Sensitivity

doi: 10.3390/ijms26188816

Figure Lengend Snippet: Capsaicin and its Antagonist AMG9810 Modulate OS Mechanotransduction by Activating or Blocking TRPV1 Channels. ( A ) Representative fluorescence microscopy images showing the effects of ASP7633 (ASP) and Capsaicin (CAP) with or without AMG (CAP+ I) on nuclear morphology. Magnification: 20×; scale bar: 25 µm. Nuclear enlargement quantification is shown as bar plots. Data represent three biological replicates, with a minimum of 30 cells per condition. ( B ) Representative confocal microscopy images of OS cells, with corresponding bar plot quantification of the nucleus-to-cytoplasm (N/C) ratio following the same treatments as in ( A ). Magnification: 20×, scale bar: 100 μm. The analysis includes three biological replicates with at least 25 cells per condition. ( C ) Migration assays showing the impact of capsaicin, with or without AMG9810, on the migratory ability of both OS cell lines. ( D ) The effect of Capsaicin treatment (with or without co-treatment with AMG9810) on the adhesion capability of U-2 OS and SAOS-2 cells. Scale bar: 100 μm. ( E ) Shows the relative cell viability of hFOB, U-2 OS, and SAOS-2 cell lines after treatment with increasing concentrations of capsaicin. ( F ) Relative cytotoxicity of U-2 OS cells treated with cisplatin or doxorubicin following capsaicin treatment with or without AMG. ( G ) Relative cytotoxicity of SAOS-2 cells treated with cisplatin or doxorubicin following capsaicin treatment with or without AMG. For each experiment, statistical analyses were performed on the most representative of three biological replicates, each comprising a minimum of four technical replicates per condition. Statistical significance between treated and control samples was determined using Student’s t -test, with significance levels indicated as p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Osteosarcoma cell lines Human SAOS-2 (HTB-85) and U-2 OS (HTB-96) and human fetal osteoblast cell line hFOB (CRL-3602) were purchased from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Blocking Assay, Fluorescence, Microscopy, Confocal Microscopy, Migration, Control

Journal: International Journal of Molecular Sciences

Article Title: A ‘Spicy’ Mechanotransduction Switch: Capsaicin-Activated TRPV1 Receptor Modulates Osteosarcoma Cell Behavior and Drug Sensitivity

doi: 10.3390/ijms26188816

Figure Lengend Snippet: TRPV1 activation modulates expression and nuclear localization Src in OS cells. ( A – D ) Representative Western blot images and their respective densitometric analyses of cell extract from U-2 OS or SAOS-2. ( A , C ) SRC (60 kDa) and GAPDH (37 kDa) protein levels following mechanical stretch or capsaicin treatment, with or without co-treatment with AMG9810, respectively. ( B , D ) Acetylated histone 3 (acH3, 17 kDa) and Histone 3 (H3 17 kDa) levels under the same conditions as above. ( E , F ) Representative immunofluorescence images of nuclei (Hoechst), Src protein, and merged channels in OS cells subjected to mechanical stretch or capsaicin treatment, with or without co-treatment with AMG9810. Src nuclear levels quantified by corrected total cell fluorescence (CTCF) are reported as bar plots (mean ± SD). ( G , H ) Western blot and CTCF analysis of Src in hFOB cells under mechanical stretch or capsaicin treatment. Densitometric analysis of the Western blot bands was performed using ImageJ 1.52 software and quantified in arbitrary units. CellProfiler 4.2.8 was used to identify nuclear SRC fluorescence intensity and area values . Data represent three biological replicates, with a minimum of 45 cells analyzed per condition. Error bars represent the standard error of the mean (SEM). Statistical analysis was conducted on at least three biological replicates. Statistical significance was determined using an unpaired t -test, with results shown as mean ± SD; p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Osteosarcoma cell lines Human SAOS-2 (HTB-85) and U-2 OS (HTB-96) and human fetal osteoblast cell line hFOB (CRL-3602) were purchased from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Activation Assay, Expressing, Western Blot, Immunofluorescence, Fluorescence, Software