Journal: Journal of cellular physiology

Article Title: The Histone-Deacetylase-Inhibitor Suberoylanilide Hydroxamic Acid Promotes Dental Pulp Repair Mechanisms Through Modulation of Matrix Metalloproteinase-13 Activity

doi: 10.1002/jcp.25128

Figure Lengend Snippet: The effects of SAHA on MMP-13 protein expression and enzyme activity. (A) Cell lysates from selected time points (1, 7, 14, and 21 days) were normalised using a Bradford dye-binding method and total MMP-13 protein levels quantified for each time point. MMP-13 production was analysed by ELISA. (B) A range of MMP-13i concentrations were added to samples of recombinant MMP-9, −13 and enzyme activity was measured fluorogenically for each inhibitor concentration. MMP-13 activity was blocked dose-dependently, but MMP-9 was not significantly altered. All experimental MMP-13i concentrations tested significantly decreased MMP-13 enzyme activity. (C/D) The effects of SAHA and a pharmacological MMP-13i on MMP-13 enzyme activity in mineralizing DPCs. DPCs were cultured in mineralizing medium±SAHA/MMP-13i and compared with mineralizing and non-mineralizing control cultures at 48 h (C) and 14 days (D). MMP-13 activity (measured fluorogenically) increased in mineralizing culture and in the presence of SAHA at both experimental time-points. As the commercial FRET substrate was preferentially, but not exclusively cleaved by MMP-13, the selective MMP-13 inhibition was used to analyse the role of MMP-13. Selective MMP-13 inhibition significantly reduced enzyme activity in the SAHA cultures. Error bars represent SEM of three independent experiments carried out in triplicate. Significant difference P<0.05, denoted by asterix (*) between experimental group and normal medium control, symbol (^) between experimental group and mineralizing medium control and symbol (‡) between mineralizing medium+SAHA and mineralizing medium+SAHA+MMP-13i.

Article Snippet: Total MMP-13 levels were analysed by quantitative sandwich ELISA technique (Cusabio, Wuhan, Hubei Province, China) according to the manufacturer’s instructions and absorbance measured at 450 nm with the correction wavelength set at 570 nm.

Techniques: Expressing, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Concentration Assay, Cell Culture, Inhibition

Journal: Journal of cellular physiology

Article Title: The Histone-Deacetylase-Inhibitor Suberoylanilide Hydroxamic Acid Promotes Dental Pulp Repair Mechanisms Through Modulation of Matrix Metalloproteinase-13 Activity

doi: 10.1002/jcp.25128

Figure Lengend Snippet: The effects of SAHA and MMP-13 activity on dental pulp cell (DPC) migration. (A) Vertical migration of DPCs measured by transwell migration assay at 37°C for 3 h. Positive control (supplemented medium and 10% FCS), 1 and 3μM SAHA had a statistically significantly greater effect on cell migration compared with the negative control. MMP-13i (1 and 2μM) and SAHA in combination with 2μM MMP-13i was not significantly different compared with the negative control, indicating the MMP-13 may be partly responsible for the migratory effects of DPCs. (B) Quantitative data from (C/D) showing cell migration in response to HDACis. (B) Horizontal migration of DPCs by wound healing scratch assay. The effect roles of 1 μm SAHA on the migration and motility of rat DPCs quantified from 24 hour scratch assay. Cells were treated with 1 μM SAHA for 24 h and a scratch made in the confluent cell monolayer. Images were obtained at 0 (Ci–iv) and 24 h post wounding (Di–iv). Values are mean±SEM for 3 independent experiments carried out in triplicate. Asterisks (*) show significant differences compared with control (P<0.05). Scale bars indicate 100μm.

Article Snippet: Total MMP-13 levels were analysed by quantitative sandwich ELISA technique (Cusabio, Wuhan, Hubei Province, China) according to the manufacturer’s instructions and absorbance measured at 450 nm with the correction wavelength set at 570 nm.

Techniques: Activity Assay, Migration, Transwell Migration Assay, Positive Control, Negative Control, Wound Healing Assay

Journal: Journal of cellular physiology

Article Title: The Histone-Deacetylase-Inhibitor Suberoylanilide Hydroxamic Acid Promotes Dental Pulp Repair Mechanisms Through Modulation of Matrix Metalloproteinase-13 Activity

doi: 10.1002/jcp.25128

Figure Lengend Snippet: Overview schematic illustration highlighting the potential roles of MMP-13 is reparative dentinogenesis. (A) HDACi (SAHA) applied topically to exposed damaged pulp tissue, accelerates mineralization processes (and the expression of a range of mineralization-associated transcripts) and increases expression of MMP-13 in pulp cells. The expression of MMP-13 is central to the control of mineralization processes and the activation of a range of bioactive molecules by cleavage. These molecules could potentially promote angiogenesis, mineralization and cell migration. Increased MMP-13 expression has also been shown in inflamed dental DP cell lines (Zhang et al., 2013), which may have additional implications for dental pup healing. (B) Dental progenitor cells migrate from vasculature in central pulp under influence of bioactive molecules to injury site. Thereafter, the progenitor cells differentiate into odontoblast-like cells. MMP-13 may directly influence this recruitment process by increasing cell migration or by cleavage of matrix-bound dentin matrix proteins to increase chemotaxis and differentiation. Additionally MMP-13 (and other MMP collagenases) will breakdown extracellular matrix further increasing migration and stimulating repair. (C) Fossilised dentine matrix proteins leach into the pulp as a result of caries (Smith et al., 2008) or restorative materials (Schröder, 1985; Tomson et al., 2007) to promote reparative events. Growth factors released by MMP-cleavage (including MMP-13) further stimulate reparative response.

Article Snippet: Total MMP-13 levels were analysed by quantitative sandwich ELISA technique (Cusabio, Wuhan, Hubei Province, China) according to the manufacturer’s instructions and absorbance measured at 450 nm with the correction wavelength set at 570 nm.

Techniques: Expressing, Activation Assay, Migration, Chemotaxis Assay

Journal: Molecular carcinogenesis

Article Title: Breast cancer malignancy is governed by regulation of the macroH2A2/TM4SF1 axis, the AKT/NF-κB pathway, and elevated MMP13 expression.

doi: 10.1002/mc.23683

Figure Lengend Snippet: FIGURE 7 mH2A2/TM4SF1 axis regulates breast cancer cell metastatic potential via the AKT/NF‐κB signaling pathway. (A) Phosphorylation of AKT/NF‐κB was evaluated in NS or shM2 T47D cells using western blot analysis. (B) Protein expression concentration was quantified using Vilber Fusion fx software and presented in a graph. (C, D) Rescue effects of TM4SF1 on AKT/NF‐κB phosphorylation were evaluated using western blot analysis. (E) Secreted MMP13 was quantified using an enzyme‐linked immunosorbent (ELISA) assay. (F) Schematic diagram suggesting the novel role of mH2A2/TM4SF1 axis regulating breast cancer metastasis through the AKT/NF‐κB signaling pathway and MM13P activity. siC, scramble siRNA; siT, siRNA against TM4SF1. (B) p Values were calculated using one‐way ANOVA with Dunnett's multiple test for comparison between the three groups. (B, D, E) p Values were calculated using one‐way ANOVA with Dunnett's multiple test for comparison between the four groups. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: MMP13 protein levels in culture supernatants were detected using a Human MMP13 ELISA kit (KE00078, Proteintech) according to the manufacturer's instructions. https://onlinelibrary.w iley.com /doi/10.1002/m c.23683 by Indian Institute O f T echnology, W iley O nline L ibrary on [24/01/2024].

Techniques: Phospho-proteomics, Western Blot, Expressing, Concentration Assay, Software, Enzyme-linked Immunosorbent Assay, Activity Assay, Comparison

Journal: Stem Cell Research & Therapy

Article Title: The differential effects of leukocyte-containing and pure platelet-rich plasma (PRP) on tendon stem/progenitor cells - implications of PRP application for the clinical treatment of tendon injuries

doi: 10.1186/s13287-015-0172-4

Figure Lengend Snippet: L-PRP induces greater catabolic responses than P-PRP. Gene expression analysis of the catabolic markers, MMP-1 and MMP-13 , by quantitative reverse transcription-polymerase chain reaction ( a ). Unlike P-PRP, L-PRP significantly induced the expression of both catabolic genes. The control group was used as the reference. Gene expression levels were normalized to the expression of GAPDH (glyceraldehyde 3-phosphate dehydrogenase). Data are represented as mean ± SD of three independent values. MMP production was also measured by using enzyme-linked immunosorbent assay ( b ). The production was normalized to respective cell number in each group. Asterisks represent significant differences when compared with the respective control, and the pound symbols indicate significant differences between L-PRP and P-PRP ( P < 0.05). Note that the data for each group were calculated from three independent values. Data are expressed as the mean ± SD. A t test was used for statistical analysis. L-PRP leukocyte-platelet-rich plasma, MMP matrix metalloproteinase, P-PRP pure-platelet-rich plasma, SD standard deviation

Article Snippet: In a separate culture, MMP-1 and MMP-13 levels in the supernatant were also measured after 4 days by using enzyme-linked immunosorbent assay (ELISA) assay kits in accordance with the instructions of the manufacturer ( MMP-1 and MMP-13 ; Biotang Inc., Boston, MA, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Marine Drugs

Article Title: Chondroitin Sulfate Nanovectorized by LC-PUFAs Nanocarriers Extracted from Salmon ( Salmo salar ) by Green Process with Decreased Inflammatory Marker Expression in Interleukin-1β-Stimulated Primary Human Chondrocytes In Vitro Culture

doi: 10.3390/md22120571

Figure Lengend Snippet: The effect of nanoliposomes/CS exposure on IL-1β-stimulated MMPs. Human chondrocytes were stimulated or not with 1 ng/mL IL-1β and exposed to nanoliposomes (250 µg/mL) or nanoliposomes and CS extracts (250 µg/mL+ 25 µg/mL). For ( A , B , E ), total RNA was extracted after 24 h stimulation then reverse transcribed into cDNA and analyzed by RT-PCR. The relative abundance of MMP1, MMP3, and MMP13 mRNAs was normalized to RP29 mRNA. Comparison was performed using the ΔCt method with the fold value of reference = 1. The results shown are the mean ± SD of at least four individual experiments (*: p < 0.01 vs. control; #: p < 0.01 vs. IL-1β). For ( C , D , F ) levels of MMP1, 3, and 13 after 48 h of stimulation (*: p < 0.01 vs. control; #: p < 0.01 vs. IL-1β).

Article Snippet: After stimulations, supernatants were collected and centrifuged 5 min at 600 g and analyses were performed according to the kit manufacturer’s instructions for MMP1, MMP3, and MMP13 levels (DuoSet ® ELISA, R&D systems, Abingdon, UK—Human Total MMP1, Human Total MMP3, and Human Pro-MMP13) and PGE2 levels (Prostaglandin E2 Parameter Assay Kit, Bio-Techne, Abingdon, UK).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Comparison, Control

Journal: Marine Drugs

Article Title: Chondroitin Sulfate Nanovectorized by LC-PUFAs Nanocarriers Extracted from Salmon ( Salmo salar ) by Green Process with Decreased Inflammatory Marker Expression in Interleukin-1β-Stimulated Primary Human Chondrocytes In Vitro Culture

doi: 10.3390/md22120571

Figure Lengend Snippet: Sequences of specific primers for RT-PCR analyses.

Article Snippet: After stimulations, supernatants were collected and centrifuged 5 min at 600 g and analyses were performed according to the kit manufacturer’s instructions for MMP1, MMP3, and MMP13 levels (DuoSet ® ELISA, R&D systems, Abingdon, UK—Human Total MMP1, Human Total MMP3, and Human Pro-MMP13) and PGE2 levels (Prostaglandin E2 Parameter Assay Kit, Bio-Techne, Abingdon, UK).

Techniques: