Journal: Bioactive Materials

Article Title: Engineering a halloysite nanotube-enhanced hydrogel 3D skin model for modulated inflammation and accelerated wound healing

doi: 10.1016/j.bioactmat.2024.11.013

Figure Lengend Snippet: Viability and functionality assessment of HFFs in HNT-modified hydrogel (a) Representative images and percentage of HFFs positive for vimentin by IF staining on days 0, 3, and 7 in natural hydrogel and 1 μg/mL HNT-based hydrogel. Data are represented as mean ± SD. ∗ P < 0.05 (two-way ANOVA analysis). Scale bar = 20 μm. (b) Relative expression levels of talin2, plasminogen, fibronectin, and paxillin assessed by RT-PCR tests in cultured HFFs on day 3. Each dot represents one independent experiment. Data are represented as mean ± SD. ns, not significant, ∗ P < 0.05; ∗∗ P < 0.01 (unpaired Student's t-test). (c) Representative images and percentage of HFFs positive for vimentin by IF staining in natural hydrogel and 1 μg/mL HNT-based hydrogel. The cells were stimulated with cytokines (TNF-α, IL-17A, and IL-22) in different concentrations (0, 25, 50, 100 ng/mL). Scale bar = 100 μm. (d) Representative 3D images from the top view of HFFs positive for vimentin in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3 and 7. The cells were stimulated without or with cytokines (TNF-α, IL-17A, and IL-22, 50 ng/mL for each cytokine). Scale bar = 100 μm. (e) Relative expression levels of vimentin in HFFs in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3, and 7 without cytokine stimulation. Data are represented as mean ± SD. ∗ P < 0.05 (two-way ANOVA analysis). (f) Relative expression levels of vimentin in HFFs in I collagen-based hydrogel, natural hydrogel, and 1 μg/mL HNT-based hydrogel on days 1, 3, and 7 with cytokine (TNF-α, IL-17A, and IL-22, 50 ng/mL for each) stimulation. Data are represented as mean ± SD. ∗∗ P < 0.01 (two-way ANOVA analysis).

Article Snippet: Subsequently, the samples were incubated overnight at 4 °C with primary antibodies, including rabbit anti-Ki-67 (catalog #34330, CST, USA), keratin (K) 5 (catalog # 71536, CST, USA), vimentin (catalog # 10366-1-AP, Proteintech, China), fibronectin (catalog # 15613-1-AP, Proteintech, China), K17 (catalog # 17516-1-AP, Proteintech, China), mouse anti-K10 (catalog # ab9025, Abcam, UK), CD34 (catalog # 31120-1-AP, Proteintech, China), or IL-17A (catalog # 66148-1-Ig, Proteintech, China).

Techniques: Modification, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture

Journal: Bioactive Materials

Article Title: Engineering a halloysite nanotube-enhanced hydrogel 3D skin model for modulated inflammation and accelerated wound healing

doi: 10.1016/j.bioactmat.2024.11.013

Figure Lengend Snippet: Inflammatory bioactivity of modified hydrogel and its potential for cell proliferation, migration, and adhesion. (a) Schematic of RNA-seq of the HFFs in hydrogel group. Four groups were set as group A (without HNT), group B (with HNT of 1 μg/mL), group C (without HNT, stimulated with TNF-α, IL-17A, and IL-22, 50 ng/mL) and group D (with HNT of 1 μg/mL, stimulated with with TNF-α, IL-17A, and IL-22, 50 ng/mL). (b) Heatmap of relative expression levels of genes associated with inflammation, proliferation and migration in group B (with HNT) compared to group A (without HNT). (c) KEGG pathway analysis of enriched genes showing the relevant functional terms in group D compared to group B. (d) Specific inflammatory clusters of genes related to KEGG analysis showing the relevant functional terms in group D compared to group B. (e) KEGG pathway analysis of enriched genes showing the relevant functional terms in group A compared to group C as well as group D compared to group B. (f) Venn diagram of differentially expressed genes overlapped in the four groups. (g) Representative 3D images of K17 expression in upper-layer from the top and side-view of the modified 3D skin model. (h) Relative expression levels of K17 in 0 μg/mL and 1 μg/mL HNT-based hydrogel with cytokine (TNF-α, IL-17A, and IL-22, 50 ng/mL) stimulation. Each dot represents one independent experiment. Data are represented as mean ± SD. ∗ P < 0.05 (unpaired student's t-test).

Article Snippet: Subsequently, the samples were incubated overnight at 4 °C with primary antibodies, including rabbit anti-Ki-67 (catalog #34330, CST, USA), keratin (K) 5 (catalog # 71536, CST, USA), vimentin (catalog # 10366-1-AP, Proteintech, China), fibronectin (catalog # 15613-1-AP, Proteintech, China), K17 (catalog # 17516-1-AP, Proteintech, China), mouse anti-K10 (catalog # ab9025, Abcam, UK), CD34 (catalog # 31120-1-AP, Proteintech, China), or IL-17A (catalog # 66148-1-Ig, Proteintech, China).

Techniques: Modification, Migration, RNA Sequencing, Expressing, Functional Assay

Journal: Parasitology Research

Article Title: Evaluation of immunoprotective effects of recombinant protein and DNA vaccine based on Eimeria tenella surface antigen 16 and 22 in vivo

doi: 10.1007/s00436-021-07105-y

Figure Lengend Snippet: Serum cytokine concentration and IgY antibody level of rEtSAG16 and 22 proteins. Serum was collected at 14 dpi (28 days of age). G1: rEtSAG16 (50 μg); G2: rEtSAG16 (100 μg); G3: rEtSAG22 (50 μg); G4: rEtSAG22 (100 μg); G5: negative control group; G6: positive control group. a IFN-γ concentration. b IgY concentration. c IL-4 concentration. d IL-10 concentration. e IL-17 concentration. Bars represent mean ± SD value ( N = 8). Different alphabet responses significant difference among groups

Article Snippet: The concentration of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-17 (IL-17) and total IgY antibody level in serum were detected through an indirect enzyme-linked immunosorbent assay (ELISA) utilising Chick Cytokine ELISA Quantitation Kits (catalogue numbers: CSB-E08550Ch, CSB-E06756Ch, CSB-E12835C, CSB-E04607Ch and CSB-E11635Ch for IFN-γ, IL-4, IL-10, IL-17 and IgY respectively; CUSABIO, Wuhan, China), according to manufacturer instructions.

Techniques: Concentration Assay, Negative Control, Positive Control

Journal: Parasitology Research

Article Title: Evaluation of immunoprotective effects of recombinant protein and DNA vaccine based on Eimeria tenella surface antigen 16 and 22 in vivo

doi: 10.1007/s00436-021-07105-y

Figure Lengend Snippet: Serum cytokine concentration and IgY antibody level of pEGFP-N1-EtSAG16, pEGFP-N1-EtSAG22 and pEGFP-N1-EtSAG4-16-22 plasmids. Serum was collected at 14 dpi (28 days of age). G7: pEGFP-N1-EtSAG16 (50 μg); G8: pEGFP-N1-EtSAG16 (100 μg); G9: pEGFP-N1-EtSAG22 (50 μg); G10: pEGFP-N1-EtSAG22 (100 μg); G11: pEGFP-N1-EtSAG4-16-22 (50 μg); G12: pEGFP-N1-EtSAG4-16-22 (100 μg); G13: empty pEGFP-N1 control; G14: negative control group; G15: positive control group. a IFN-γ concentration. b IgY concentration. c IL-4 concentration. d IL-10 concentration. e IL-17 concentration. Bars represent mean ± sd value ( N = 8). Different alphabet responses significant difference among groups

Article Snippet: The concentration of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-17 (IL-17) and total IgY antibody level in serum were detected through an indirect enzyme-linked immunosorbent assay (ELISA) utilising Chick Cytokine ELISA Quantitation Kits (catalogue numbers: CSB-E08550Ch, CSB-E06756Ch, CSB-E12835C, CSB-E04607Ch and CSB-E11635Ch for IFN-γ, IL-4, IL-10, IL-17 and IgY respectively; CUSABIO, Wuhan, China), according to manufacturer instructions.

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