Journal: bioRxiv

Article Title: Induction of cervical dysfunction associated with preterm birth by IL-1 and dysbiotic microbiome revealed in human endocervix chips

doi: 10.1101/2025.05.01.651107

Figure Lengend Snippet: A) Bar graph showing total colony forming units (CFUs)/Chip of BVC1 present in the inoculum and detected on Endocervix versus Ectocervix Chips 96 hours post-inoculation (N=6 chips per group, 2 independent experiments for each tissue type with each indicated by ○ or □ (Mean ± SD, log10 scale, one-way ANOVA). B) Immunofluorescence microscopic side-view images (left) of Endocervix Chips inoculated with either L. crispatus or BVC1 showing bacteria (purple, Bactoview) and mucus (green, WGA) (Bar, 200 μm) with bar graph at right depicting WGA lectin intensity (N=3-4 chips Mean ± SD, one-way ANOVA). C) Bar graphs showing cytokine (IL-1β, IL-6, IL-8, TNF-α, CXCL-10) and MMP-8 levels in apical outflows of Endocervix Chips 72 hours post-inoculation with the L. crispatus or BVC1 consortium (N=10 chips; 3 donors with results from each labeled as ○, □ or Δ; Mean ± SD, t -test).

Article Snippet: To assess the effect of inflammatory cytokines on mucus production, live chips were removed from Pods and exposed apically to 150μl of the lectin Ulex Europaeus Agglutinin I (UEA I), Rhodamine (Vector Laboratories, RL-1062-2) resuspended in HBSS++ at 10μg/ml (1:200 dilution) for 20 minutes at 37°C by gravity flow followed by two gravity washes with 150μl HBSS++.

Techniques: Immunofluorescence, Bacteria, Labeling

Journal: bioRxiv

Article Title: Induction of cervical dysfunction associated with preterm birth by IL-1 and dysbiotic microbiome revealed in human endocervix chips

doi: 10.1101/2025.05.01.651107

Figure Lengend Snippet: A) Immunofluorescence microscopic top-view images of the epithelium within Endocervix Chips showing Ulex Europaeus Agglutinin I (UEA I)-stained mucus after 24 hours without (control) or with exposure to a cytokine mixture of IL-1β, IL-6, IL-8 & TNF-α (bar, 180 μm) and bar graph at the right showing quantification of fluorescence intensities (-, control; +, with cytokines; 6 non-overlapping fields of view; N=3 chips; Mean ± SD, t -test). B) Graph showing levels MUC5B and MUC5AC gene expression determined by qRT-PCR in Endocervix Chip epithelia 24 hours after perfusion with cytokine mixture shown in A (N=6 chips, 2 donors, 2 independent repeats, Mean ± SD, t -test). Data were normalized to housekeeping genes and control chips (ΔΔCt). C) Immunofluorescence microscopic images of the Endocervix Chip stroma stained with F-actin (magenta) and Hoechst (nuclei, cyan) (bar, 100 μm). D) Graph showing levels of PTGS2 , PGR , and COL1A1 gene expression in PAH-perfused Endocervix Chip stroma with (+) and without (-) cytokine treatment (N=6-10 chips, 2-3 donors with results from each donor labeled as ○, □ or Δ;). Data normalized to housekeeping genes and control chips (ΔΔCt, Mean ± SD, t -test). E) Representative second harmonic microscopic images (left) of fibrillar collagen within the stroma in the basal channel of control and cytokine-perfused Endocervix Chips (bar, 100 μm) with quantification shown in the bar graph at the right (-, control; +, with cytokines; 9-10 non-overlapping fields of view, N=3 chips, Mean ± SD, t -test). F) Gene expression levels expressed as count per million (CPM)-normalized counts of IL-8 ( CXCR1 , CXCR2 ), IL-1 ( IL1R1 , IL1R2 ), IL-6 ( IL6R ), and TNF-α ( TNFRSF1A , TNFRSF1B ) receptors in the Endocervix Chip epithelium (white bars) and stroma (light grey). (N=10 chips, 3 donors, Mean ± SD, Two-way ANOVA with multiple comparisons).

Article Snippet: To assess the effect of inflammatory cytokines on mucus production, live chips were removed from Pods and exposed apically to 150μl of the lectin Ulex Europaeus Agglutinin I (UEA I), Rhodamine (Vector Laboratories, RL-1062-2) resuspended in HBSS++ at 10μg/ml (1:200 dilution) for 20 minutes at 37°C by gravity flow followed by two gravity washes with 150μl HBSS++.

Techniques: Immunofluorescence, Staining, Control, Fluorescence, Gene Expression, Quantitative RT-PCR, Labeling