Journal: International Wound Journal

Article Title: Curcumin accelerates cutaneous wound healing via multiple biological actions: The involvement of TNF‐α, MMP‐9, α‐SMA, and collagen

doi: 10.1111/iwj.12904

Figure Lengend Snippet: Curcumin down‐regulated MMP‐9 expression in TNF‐α‐treated Hs68 cells via NF‐κB signal pathway. (A) Hs68 cells were incubated with the indicated concentrations of curcumin for 1 hour and then with 3 ng/mL TNF‐α for 23 hours in the continued presence of curcumin. MMP‐9 expression was analysed by immunofluorescence staining. Scale bar = 100 μm. (B) MMP‐9 protein in cell lysates was measured by Western blot. GAPDH was used as the loading control. The data are expressed as a fold value compared with the control value and are shown as the mean ± SD for 3 separate experiments. (C) Western blot analysis for the phosphorylation of NF‐κB p65. Hs68 cells were pre‐incubated for 1 hour with 20 μM curcumin and were then treated with 3 ng/mL TNF‐α for 5 minutes. (D) Immunofluorescence staining for NF‐κB p65. Hs68 cells were pre‐incubated for 1 hour with 20 μM curcumin and were then treated with 3 ng/mL TNF‐α for 30 minutes. Representative results from 3 separate experiments are shown. (E) Cells were co‐incubated for 1 hour with 0–20 μM Bay11–7082 (a NF‐κB inhibitor) and then with 3 ng/mL TNF‐α for 23 hours. Cell lysates were prepared and assayed for MMP‐9 by Western blot. The data are expressed as mean ± SD for 3 separate experiments. *P < .05 compared with the untreated cells. †P < .05 compared with the TNF‐α‐treated cells

Article Snippet: Hs68 cells were pre‐treated with different concentrations of curcumin (0‐2.5 μM) and an MMP‐9 inhibitor (CAS 1177749584;5 μM; Santa Cruz Biotechnology) for 1 hour.

Techniques: Expressing, Incubation, Immunofluorescence, Staining, Western Blot

Journal: International Wound Journal

Article Title: Curcumin accelerates cutaneous wound healing via multiple biological actions: The involvement of TNF‐α, MMP‐9, α‐SMA, and collagen

doi: 10.1111/iwj.12904

Figure Lengend Snippet: Curcumin increases myofibroblast differentiation via the inhibition of NF‐κB expression. (A) Immunohistochemical staining for α‐SMA expression in control and curcumin‐treated wounds at the determined time. The marked area in the upper panel was shown in the lower panel at a higher magnification. α‐SMA‐positive myofibroblasts were increased in the curcumin‐treated group than the control group at postoperative day 7 and 12. The arrowheads indicate myofibroblasts. Scale bar: upper = 1 mm; lower = 100 μm. (B) Hs68 cells were incubated for 1 hour with a different concentration of curcumin; then, the cells were incubated with 3 ng/mL of TNF‐α for 23 hours. α‐SMA expression was analysed by immunofluorescence staining. Scale bar = 50 μm. (C) Hs68 cells were incubated with the indicated concentration of curcumin for 1 hour and then with 3 ng/mL TNF‐α for 23 hours in the continued presence of curcumin; α‐SMA protein in cell lysates was then measured by Western blot. GAPDH was used as the loading control. (D) Cells were co‐incubated for 1 hour with 0–20 μM Bay11–7082 (a NF‐κ B inhibitor) and then with 3 ng/mL TNF‐α for 23 hours. Cell lysates were prepared and assayed for α‐SMA by Western blot. The data are expressed as a fold value compared with the control value and are shown as the mean ± SD for 3 separate experiments. *P < .05 compared with the untreated cells. †P < .05 compared with the TNF‐α‐treated cells

Article Snippet: Hs68 cells were pre‐treated with different concentrations of curcumin (0‐2.5 μM) and an MMP‐9 inhibitor (CAS 1177749584;5 μM; Santa Cruz Biotechnology) for 1 hour.

Techniques: Inhibition, Expressing, Immunohistochemical staining, Staining, Incubation, Concentration Assay, Immunofluorescence, Western Blot

Journal: International Wound Journal

Article Title: Curcumin accelerates cutaneous wound healing via multiple biological actions: The involvement of TNF‐α, MMP‐9, α‐SMA, and collagen

doi: 10.1111/iwj.12904

Figure Lengend Snippet: Curcumin increases collagen production in wound and in TNF‐α‐treated fibroblasts. (A) The sections with Masson's trichrome staining of wound were scanned with an Aperio CS2 digital pathology scanner. The dotted marked area in the upper panel was shown in the lower panel at higher magnification. Scale bar: upper = 3 mm; lower = 1 mm. (B) The scoring system was used to evaluate collagen deposition (blue colour) by Masson's trichrome staining, using a 5‐point visual scoring scale. The collagen deposition score (grade 5) in the curcumin‐treated group was significantly greater than the control. *P < .05 compared with the control group at the determined time. (C) The level of collagen protein was measured by Western blot. Hs68 cells were incubated with the indicated concentrations of curcumin for 1 hour and then with 3 ng/mL TNF‐α for 23 hours in the continued presence of curcumin; collagen protein in cell lysates was then measured by Western blot. GAPDH was used as the loading control. The data are expressed as a fold value compared with the control value and are shown as the mean ± SD for 3 separate experiments. *P < .05 compared with the untreated cells. †P < .05 compared with the TNF‐α‐treated cells. (D) The total amount of collagen was measured by Sircol collagen assay. Hs68 cells were incubated with the MMP‐9 inhibitor (5 μM) and with the indicated concentrations of curcumin for 1 hour and then with 3 ng/mL TNF‐α for 23 hours. The supernatant was collected, and the level of collagen was measured by Sircol collagen assay. *P < .05 compared with the TNF‐α‐treated cells. †P < .05 compared with the TNF‐α+MMP9 inhibitor‐treated cells. ǂP < .05 compared with the TNF‐α+curcumin‐treated cells

Article Snippet: Hs68 cells were pre‐treated with different concentrations of curcumin (0‐2.5 μM) and an MMP‐9 inhibitor (CAS 1177749584;5 μM; Santa Cruz Biotechnology) for 1 hour.

Techniques: Staining, Western Blot, Incubation, Sircol Collagen Assay

Journal: Oncotarget

Article Title: Tumor suppressor REIC/DKK-3 and co-chaperone SGTA: Their interaction and roles in the androgen sensitivity

doi: 10.18632/oncotarget.6488

Figure Lengend Snippet: A. The yeast two-hybrid analysis was conducted using pPC86 (AD)/full-length human SGTA (derived from a normal heart cDNA library) and pDBLeu (BD)/full-length human REIC/DKK-3 plasmids. The blue colonies indicate those with an interaction between the two proteins. B. For the pull-down (PD) assay, the full-length cDNA of human REIC/DKK-3 and SGTA was cloned into the pFN21A and pMACS Kk.HA-C plasmids, respectively. Cell lysates from Halo-tagged REIC/DKK-3- and/or HA-tagged SGTA-transfected 293T cells were analyzed. The sample pulled down using Halo-tagged REIC/DKK-3 was analyzed by Western blotting (WB) using anti-HA antibody. C. REIC/DKK-3 and SGTA protein expression in 293T, PC3 and Hs68 cells was analyzed by Western blotting. Coomassie Brilliant Blue (CBB) staining of the membrane is shown as a loading control. D. The co-localization of REIC/DKK-3 and SGTA was examined by double immunofluorescence staining and observed by fluorescence microscopy. The images in green and red show the intracellular localization of REIC/DKK-3 and SGTA, respectively. The areas of overlap between REIC/DKK-3 and SGTA are shown in yellow in the merged image.

Article Snippet: The normal human foreskin fibroblast cell line Hs68 was provided by JCRB Cell Bank (Osaka, Japan).

Techniques: Derivative Assay, cDNA Library Assay, Clone Assay, Transfection, Western Blot, Expressing, Staining, Membrane, Control, Double Immunofluorescence Staining, Fluorescence, Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Adipose-derived stem cell spheroid-laden microbial transglutaminase cross-linked gelatin hydrogel for treating diabetic periodontal wounds and craniofacial defects

doi: 10.1186/s13287-023-03238-2

Figure Lengend Snippet: In vitro characterization of ADSCs and ADsp-mTG. A Pluripotent gene expression in ADSCs after 24 h of seeding. B The trilineage differentiation of ADSCs by Oil Red O (adipogenesis), Alizarin Red (osteogenesis), and Alcian Blue (chondrogenesis) at day 14. Scale bar: 100 µm. C The spreading of ADSCs from spheroids embedded in mTG. D The trilineage differentiation of ADsp in mTG at day 14. Scale bar: 100 µm. There is no downstream processing or averaging to adjust the resolution of the microscopic images. (Significant difference compared to Hs68: * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: Hs68 (cell line) , Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: In Vitro, Gene Expression

Journal: Stem Cell Research & Therapy

Article Title: Adipose-derived stem cell spheroid-laden microbial transglutaminase cross-linked gelatin hydrogel for treating diabetic periodontal wounds and craniofacial defects

doi: 10.1186/s13287-023-03238-2

Figure Lengend Snippet:

Article Snippet: Hs68 (cell line) , Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Isolation, cDNA Synthesis, Staining, Polymer

Journal: Pharmaceutics

Article Title: Comprehensive Analysis of the Safety Profile of a Single-Stranded RNA Nano-Structure Adjuvant

doi: 10.3390/pharmaceutics11090464

Figure Lengend Snippet: Dose-dependent cell viabilities of Hs68 cell line treated with the ssRNA nano-structure adjuvant, using MTT assays. Relative viabilities of Hs68 cells were compared to negative control (0 concentration of ssRNA nano-structure adjuvant) from 24 h to 72 h, based on the ssRNA concentration (20 and 200 μg). Poly I:C (20 and 200 μg) was used as a positive control. Unlike poly I:C, the ssRNA did not affect cell viability in Hs68 cells. The data were normalized to 100%. The data shown are expressed as the mean ± SD.

Article Snippet: HS68 cells, derived from human foreskin fibroblast were obtained from Korean Cell Line Bank (Seoul, Korea).

Techniques: Adjuvant, Negative Control, Concentration Assay, Positive Control