Journal: International journal of oncology

Article Title: Contribution of the PI3K/MMPs/Ln-5γ2 and EphA2/FAK/Paxillin signaling pathways to tumor growth and vasculogenic mimicry of gallbladder carcinomas.

doi: 10.3892/ijo.2013.1897

Figure Lengend Snippet: Figure 3. Expression of MMP-2, MT1-MMP proteins from sections [(A) SABC method, original magnification, x200] and supernates [(B) ELISA] of 3-D culture samples in vitro in GBC-SD, SGC-996 and GBC-SD+TIMP-2 groups. (A) The positive expression site of MMP-2 and MT1-MMP proteins presented yellow-brown reactant in the cytoplasm. Overexpression of MMP-2 (a1) and MT1-MMP (a4) proteins in GBC-SD group was observed. Expression of MMP-2 and MT1-MMP proteins in SGC-996 group (a2,5) and GBC-SD+TIMP-2 (a3,6) group was significantly decreased (*P<0.001, #P<0.01, vs. GBC-SD group). (B) Expression of MMP-2 (b1) and MT1-MMP (b2) proteins in GBC-SD group increased significantly with time, when compared with SGC-996 group and GBC-SD+TIMP-2 group (*P<0.001).

Article Snippet: The sections (4-μm) from each group were dehydrated in xylene and graded ethanol series, were added in order with primary antibody [MMP-2 (1: 200), MT1-MMP (1:100); rabbit polyclonal antibody, Wuhan Boster Co., China)], biotinylated secondary antibody, SABC reagents and DAB solution (Wuhan Boster Co.), respectively.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, In Vitro, Over Expression

Journal: International journal of oncology

Article Title: Contribution of the PI3K/MMPs/Ln-5γ2 and EphA2/FAK/Paxillin signaling pathways to tumor growth and vasculogenic mimicry of gallbladder carcinomas.

doi: 10.3892/ijo.2013.1897

Figure Lengend Snippet: Figure 4. Expression of MMP-2, MT1-MMP proteins/mRNAs from sections of tumor xenografts in vivo [(A) SABC method, original magnification, x200; (B) qRT-PCR)] in GBC-SD, SGC-996 and GBC-SD+TIMP-2 groups. Overexpression of MMP-2 (Aa1,7) and MT1-MMP (Aa4,7) proteins or mRNA (B) in GBC-SD group was observed in vivo. Expression of MMP-2 and MT1-MMP proteins or mRNA in SGC-996 group (Aa2,5,7 and B) and GBC-SD+TIMP-2 (Ac3,6,7 and B) group was significantly decreased (*P<0.001, vs. GBC-SD group).

Article Snippet: The sections (4-μm) from each group were dehydrated in xylene and graded ethanol series, were added in order with primary antibody [MMP-2 (1: 200), MT1-MMP (1:100); rabbit polyclonal antibody, Wuhan Boster Co., China)], biotinylated secondary antibody, SABC reagents and DAB solution (Wuhan Boster Co.), respectively.

Techniques: Expressing, In Vivo, Quantitative RT-PCR, Over Expression

Journal: Biomaterials

Article Title: Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

doi: 10.1016/j.biomaterials.2016.05.011

Figure Lengend Snippet: Schematic diagrams of the peptide–functionalized Scl2 protein and hydrogel. (A) Schematic diagram of the Scl2 protein containing H–binding, I–binding, and HA–binding peptide sequences. The ‘CPPC’ domains represent the amino acid sequences inserted at the N and C termini to aid stability of each construct. (B) Generalized structure of the peptide–functionalized Scl2 protein and hydrogel. For clarity, the homotrimer chains are represented as different colors, and the triple helical motif has been opened out to show the three chains. The locations of the H–binding and I–binding sites are not proportional to their locations in the construct. The 2–arm acrylate–functionalized MMP7 and ACAN degradable peptides were conjugated to the Scl2 protein via thiol–acrylate reactions to generate hydrogels. (C) Schematic diagrams of different Scl2 protein hydrogel samples.

Article Snippet: MMP7 and ADAMTS4 fluorogenic substrates were purchased from Merck Millipore (UK).

Techniques: Binding Assay, Construct

Journal: Biomaterials

Article Title: Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

doi: 10.1016/j.biomaterials.2016.05.011

Figure Lengend Snippet: Degradation profile of acellular Scl2 hydrogels. Degradation of hydrogels incubated in chondrogenic medium supplemented with 30 ng/mL recombinant human (A) MMP7 or (B) ADAMTS4 over time characterized as dry weight loss and expressed as a percentage of initial dry weight. Degradation of hydrogels cross–linked via (C) MMP7 or (D) ADAMTS4 degradable peptides and exposed to 30 ng/mL recombinant human MMP1, 2, 7, or 13 over time characterized as dry weight loss and expressed as a percentage of initial dry weight. Trypsin–driven degradation is used as a positive control. Chondrogenic medium without exogenous enzymes is used as a negative control. Values represent means ± SD. *** p < 0.001 versus MMP7:ACAN(25:75)-HIHA-Scl2 (n = 3).

Article Snippet: MMP7 and ADAMTS4 fluorogenic substrates were purchased from Merck Millipore (UK).

Techniques: Incubation, Recombinant, Positive Control, Negative Control

Journal: Biomaterials

Article Title: Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

doi: 10.1016/j.biomaterials.2016.05.011

Figure Lengend Snippet: Metabolic activity and DNA content of hMSCs cultured in Scl2 hydrogels. (A) hMSC metabolic activity in hydrogels over 6 weeks in culture. (B) DNA content of hMSCs per construct in hydrogels cultured over 6 weeks in culture. All data normalized to day 0. Values represent means ± SD. * p < 0.05 versus MMP7:ACAN(25:75)-HIHA-Scl2 (n = 3 for each donor; 3 different bone marrow–derived hMSC donors).

Article Snippet: MMP7 and ADAMTS4 fluorogenic substrates were purchased from Merck Millipore (UK).

Techniques: Activity Assay, Cell Culture, Construct, Derivative Assay

Journal: Biomaterials

Article Title: Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

doi: 10.1016/j.biomaterials.2016.05.011

Figure Lengend Snippet: ECM accumulation and compressive modulus of hMSC–seeded Scl2 hydrogels. (A) Sulfated glycosaminoglycan (sGAG) content of tissue deposited by hMSCs in hydrogels over 6 weeks in culture. (B) Hydroxyproline content of tissue deposited by hMSCs in hydrogels over 6 weeks in culture as an estimation of total collagen content. (C) Dynamic mechanical analysis (DMA) used to determine the elastic modulus of compression of hMSC–seeded hydrogels compressed to 10% strain at 0.5% strain/min and 0.1–10 Hz after 6 weeks in culture. Values represent means ± SD. * p < 0.05, ** p < 0.01, ## p < 0.01 versus MMP7:ACAN(25:75)-HIHA-Scl2, *** p < 0.001 (n = 3 for each donor; 3 different bone marrow–derived hMSC donors). (D) Representative histological and immunohistochemical examination of hMSC–seeded hydrogels after 6 weeks in culture. Hydrogels are stained with Haematoxylin and Eosin (H&E), Alcian Blue for sGAG, and by immunohistochemistry for collagen type I, collagen type II, and collagen type X, respectively, from top to bottom. Scale bars are 50 μm.

Article Snippet: MMP7 and ADAMTS4 fluorogenic substrates were purchased from Merck Millipore (UK).

Techniques: Derivative Assay, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Biomaterials

Article Title: Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

doi: 10.1016/j.biomaterials.2016.05.011

Figure Lengend Snippet: hMSC gene expression and enzymatic activity in Scl2 hydrogels. (A) MMP7, (B) ADAMTS4, (C) TIMP2, and (D) TIMP3 gene expression of hMSCs encapsulated in hydrogels over 6 weeks in culture, as analyzed using the ΔΔCt method. Data presented as a fold difference relative to undifferentiated hMSCs (calibrator) prior to encapsulation and normalized to GAPDH (housekeeping gene). Activity of (E) MMP7 and (F) ADAMTS4 in hydrogels over 6 weeks in culture. MMP7 and ADAMTS4 activities were normalized to fluorescence signal output at day 0. Values represent means ± SD. * p < 0.05, # p < 0.05 versus MMP7:ACAN(25:75)-HIHA-Scl2, ** p < 0.01, ## p < 0.01 versus MMP7:ACAN(25:75)-HIHA-Scl2, *** p < 0.001, ### p < 0.001 versus MMP7:ACAN(25:75)-HIHA-Scl2 (n = 3 for each donor; 3 different bone marrow–derived hMSC donors).

Article Snippet: MMP7 and ADAMTS4 fluorogenic substrates were purchased from Merck Millipore (UK).

Techniques: Gene Expression, Activity Assay, Encapsulation, Fluorescence, Derivative Assay