Journal: Acta biomaterialia

Article Title: Assessing Bioactivity and Biointegration of Engineered Salivary Tissue Constructs in a Preclinical Unilateral Fractionated Irradiated Rat Model

doi: 10.1016/j.actbio.2026.09.027

Figure Lengend Snippet: (A) H&E staining on right parotid glands of irradiated (IR) and age-matched non-irradiated (non-IR) rats at each collected timepoint. Scale bar, 50 μm. (B) Masson’s Trichrome staining on right parotid glands of non-IR and IR rats at each timepoint. Scale bar, 200 μm. (C) Quantification of acinar cell number, acini area, and level of fibrosis detected in tissues from A-B. Mean ± SEM. *, P < 0.05, ***, P < 0.001, ****, P < 0.0001. (D) Right parotid gland from non-IR and IR animals at each timepoint immunostained for α-smooth muscle actin (ACTA2, red), Aquaporin 5 (AQP5, green), Fatty Acid Binding Protein 4 (FABP4, green), β-III Tubulin (TUBB3, red), and DAPI (grey). Scale bars, 20 μm. (E) Quantification of total acinar cell-related AQP5 protein, myoepithelial cell-related ACTA2 protein, endothelial cell-related FABP4 protein, and neuronal cell-related TUBB3 protein expression. Mean ± SEM. *, P < 0.05. **, P < 0.01.

Article Snippet: The primary antibodies used were E-cadherin (13–1900, Invitrogen), AQP5 (AQP-005, Alamone Labs), α-Smooth Muscle Actin (ACTA2) (A2547, Sigma-Aldrich), β-III Tubulin (TUBB3) (MAB1195, R&D Systems), FABP4 (AF1443, R&D Systems), Ki-67 (550609, BD Biosciences), Lamin A/C ( AB133256 , Abcam), Cleaved Caspase 3 (9661, Cell Signaling Technology), Cleaved PARP (PARP) (9148, Cell Signaling Technology), α-Amylase (A8273, Sigma-Aldrich), RECA-1 (MCA970GA, Biorad), and KRT19 (Troma-III, DHSB).

Techniques: Analysis, Staining, Irradiation, Binding Assay, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of lack of salivation on the oral and cecal microbiome. ( A ) Expression of Aqp5 protein in WT and Aqp5 -/- submandibular glands obtained from 5 mice was assayed by Western blot. ( B ) Pilocarpine-stimulated saliva secretion in WT and Aqp5 -/- mice. Here and in all panels, each dot represents 1 mouse. ( C ) Heatmaps of the S16 analysis of the oral microbiome. Full list in accession number: PRJNA1260208 . ( D–I ) Analysis of the WT ( black and blue ) or Aqp5 -/- mice ( red ) oral microbiomes: similarities ( D ); observed OTUs ( E ); Chao1 index ( F ); Ace index ( G ); Shannon index ( H ), and Simpson index ( I ). ( J ) Analysis of phyla in the oral microbiome of WT and Aqp5 -/- mice. ( K ) Analysis of genus in the oral microbiome of WT and Aqp5 -/- mice. ( L ) Heatmaps of the S16 analysis of the cecal microbiome. Full list in accession number: PRJNA1260208 . ( M–Q ) Analysis of the WT ( black and blue ) or Aqp5 -/- mice ( red ) cecal microbiomes: similarities ( M ), Chao1 index ( N ), Shannon index ( O ); Simpson index ( P ); observed OTUs ( Q ). ( R ) Analysis of phyla in the cecal microbiome of WT and Aqp5 -/- mice. ( S ) Analysis of genus in the cecal microbiome of WT and Aqp5 -/- mice. ( T ) Level of intestinal neutrophils in WT and Aqp5 -/- mice.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome, Expressing, Western Blot, Analysis

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Changes in the cecal microbiome and inflammatory mediators in mice with reduced saliva due to deletion of Aqp5 and treated with DSS. ( A–G ) Analysis of cecal genus spices in the microbiome of WT and Aqp5 -/- mice. ( H–S ) Measurement of inflammatory mediators in the colon ( H–M ) and the serum ( N–S ) of WT and Aqp5 -/- mice treated with DSS for 6 days. Each dot represents 1 mouse.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome, Analysis

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of lack of saliva on development of DSS-induced colitis. ( A ) WT ( black and green ) and Aqp5 -/- mice ( blue and red ) were fed with 3% DSS ( green and red ) for 15 days. Mice were 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are the difference between WT and Aqp5 -/- mice fed with DSS. ( B ) Time course of development of disease activity of the mice in ( A ). ( C and D ) WT and AQP5 -/- mice age 8 ( C ) and 13 ( D ) weeks were treated with 3% DSS added to drinking water. The number of mice in each group is indicated in parenthesis, and the results are mean ± SEM. The P values are relative to WT mice fed with DSS. ( E ) Intestinal permeability expressed as fold change relative to the permeability measured before DSS treatment started and was taken as 1 for each group of mice. ( F ) Example images and averages of intestinal length. ( G–I ) Example images ( G ) and the level of ZO1 ( H ) and Occludin ( I ) measured by immunofluorescence staining. ( J–L ) Example H&E and PAS images ( J ) and analysis of cell proliferation ( K ) and goblet cell density ( L ). ( M and N ) Serum ( M ) and intestinal MPO ( N ) levels as reporters of neutrophils activity. ( O and P ) WT and AQP5 -/- mice age 18 weeks (5 mice in each group) were treated with 3% DSS and were gavaged with 50 μL PBS ( Black, blue, gray and green ) or with 50 μL whole saliva collected from 3 WT mice by stimulation with pilocarpine. The mice were gavaged on days 0, 3, and 6. Mice weight ( O ) was measured in 2-day intervals, and intestinal permeability ( P ) was measured on days 1, 3, 6, and 12. The results are mean ± SEM.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Activity Assay, Permeability, Immunofluorescence, Staining, Analysis, H&E Stain

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of saliva fluid and microbiome fraction on development of DSS-induced colitis. ( A ) Illustration of the protocol and procedures used to collect the salivary microbiome and fluid fractions. ( B and D ), WT ( B ) and Aqp5 -/- mice ( D ) were fed with water ( black ) or 3% DSS ( blue, green and red ) for 6 days and allowed to recover for 9 days. The mice were gavaged with PBS ( black ), the microbiome fraction in PBS ( green ), or the salivary fluid fraction ( red ) on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. ( C and E ) Time course of development of disease activity of the mice in ( B and D ), respectively. ( F ) Intestinal permeability expressed as fold change measured on day 8. ( G ) Example images and averages of intestinal length measured on day 8. ( H–J ) Example immunofluorescence and bright field images ( H ) and analysis of ZO1 ( I ) and Occludin ( J ) measured on day 8. ( K–M ) Example H&E and PAS images ( K ) and analysis of cell proliferation ( L ) and goblet cell density ( M ). ( N and O ) Serum ( N ) and intestinal MPO ( O ) levels.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome, Activity Assay, Permeability, Immunofluorescence, Analysis

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of the separated WT microbiome and salivary fluid on the serum and colonic inflammatory mediators in WT and Aqp5 -/- mice. Inflammatory mediators were measured at days 8 and 15 of WT ( open columns ) and of Aqp5 -/- mice ( filled columns ) treated with DSS ( blue ) and gavaged with WT microbiome fraction ( green ) of fluid fraction ( red ).

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of the WT microbiome and salivary fluid on the microbiome of WT and Aqp5 -/- mice. ( A and B ) Heatmaps of the WT and Aqp5 -/- mice microbiomes treated with DSS and the microbiome or fluid fractions. Full list in accession number: PRJNA1260208 . ( C–F ) Analysis of the cecal microbiome of WT ( open symbols and open columns ) and of Aqp5 -/- mice ( closed symbols and filled columns ) treated with DSS ( red, green and blue ) and gavaged with WT microbiome ( green ) of fluid fraction ( red ). ( G and H ) Analysis of Phyla and Genus level of the various mice. ( I–N ) Analysis of selective cecal genus spices in the microbiome of WT and Aqp5 -/- mice treated with DSS and gavaged with the microbiome of fluid fraction obtained from WT mice, as indicated.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome, Analysis

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of saliva fluid and microbiome fractions on development of DSS-induced colitis in the absence of the resident microbiome. ( A and D ) WT ( A ) and Aqp5 -/- mice ( D ) were treated with broad spectrum antibiotics for 1 week to clear the resident microbiome prior to the start of the experiment. The mice were fed with water ( black ) or 3% DSS ( blue, green and red ) for 6 days and allowed to recover for 6 days. The mice were gavaged with PBS ( black ), the microbiome fraction in PBS ( green ), or the salivary fluid fraction ( red ) on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. ( B and E ) Time course of development of disease activity of the mice in ( A and D ), respectively. ( C and F ) Survival curves of WT ( C ) and Aqp5 -/- mice ( F ). P values between the indicated groups are shown in the Figures. ( G ) Intestinal permeability expressed as fold change measured on day 8.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Microbiome, Activity Assay, Permeability

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Effect of saliva fluid fractions, MIF, and TFF2 on development of DSS-induced colitis. ( A ) Coomassie-stained proteins in whole saliva fluid, ( 1 ) >100-kDa fraction, ( 2 ) >50-kDa fraction, ( 3 ) >30-kDa fraction, and ( 4 ) <30-kDa fraction. ( B and C ), WT ( B ) and Aqp5 -/- mice ( C ) were fed with water ( black ) or 3% DSS ( blue, green, red and dark yellow ) for 6 days and allowed to recover for 9 days. The mice were gavaged with the indicated fractions of the salivary fluid on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. The results with DSS shown in dotted lines were taken from and are shown to better illustrate the effect of the fractions. ( D ) Venn chart of the proteins in each fraction. ( E ) A list of the proteins in the 50-KDa fraction. MIF and TFF2 are highlighted in red . ( F and G ) Protocol as in ( B and C ), except that the mice were gavaged with 50 μL of 0.01 μg/μL MIF ( green ) or 0.01 μg/μL TFF2 ( red ). ( H and I ) Example images ( H ) and averages of intestinal length ( I ) measured on day 8. ( J–L ) Example H&E images ( J ) and analysis of cell proliferation ( K ) and goblet cell density ( L ). ( M ) Intestinal permeability expressed as fold change measured on day 8. ( N–P ) Example of immunofluorescence and bright field images ( N ) and analysis of ZO1 ( O ) and Occludin ( P ) measured on day 8. ( Q and R ) Intestinal ( Q ) and serum ( R ) MPO levels.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Staining, Analysis, Permeability, Immunofluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Role of Innate Oral Immunity and the Salivary Fluid in Inflammatory Bowel Disease

doi: 10.1016/j.jcmgh.2025.101713

Figure Lengend Snippet: Scavenging saliva fluid MIF and TFF2 ameliorates effects of saliva fluid on development of DSS-induced colitis. ( A and B ) Standard curves ( left ) and the level of MIF ( A ) and TFF2 ( B ) in saliva from control ( black ) and DSS-treated mice ( red ) and saliva treated with the respective antibodies ( green ). ( C and E ) WT ( C ) and Aqp5 -/- mice ( E ) were fed with water ( black ) or 3% DSS ( blue, green and red ) for 6 days and allowed to recover for 6 days. The mice were gavaged with the salivary fluid treated with anti-MIF ( green ) and anti-TFF2 antibodies ( red ) on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. The results with DSS shown in dotted lines were taken from and are shown to better illustrate the effect of MIF and TFF2 depletion. ( D and F ) Time course of development of disease activity of the mice in ( C and E ), respectively. ( G ) Intestinal length measured on day 8. ( H and I ) Analysis of cell proliferation ( left ) and goblet cell density ( right ). ( J and K ) Intestinal ( J ) and serum ( K ) MPO levels. ( L ) Intestinal permeability expressed as fold change measured on day 8. ( M and N ) Analysis of ZO1 ( M ) and Occludin ( N ) measured on day 8.

Article Snippet: Aqp5 was detected by 1:100 dilution of anti-Aqp5 antibodies (Alomone Labs, AQP-005Cat).

Techniques: Control, Activity Assay, Analysis, Permeability, H&E Stain