type i collagen hydrogel  (Advanced Biomatrix Inc)

Journal: bioRxiv

Article Title: A human synovial tendon-on-a-chip models key features of peritendinous adhesions and offers a new approach methodology for testing anti-fibrotic drugs

doi: 10.64898/2026.04.03.716316

Figure Lengend Snippet: a , Culture timeline outlining the steps to incorporate a synovial layer. A tendon construct containing primary tendon fibroblasts and macrophages was matured in parallel in the bottom chamber. FLS are encapsulated in a type I collagen hydrogel and added to the basal side of the porous nanomembrane (acellular collagen for - FLS), followed by seeding vascular endothelial cells on the apical side to form the vascularized synovial compartment. At Day 0, the vascular-synovial compartment is joined with the bottom tendon compartment. Freshly isolated monocytes are added to the top compartment with or without TGF-β1 in the bottom compartment. Experimental analyses are performed from Day 1 to Day 5. b, Schematic detailing the functional unit of an intrasynovial tendon that is recapitulated in the synToC. The confocal immunofluorescence image corresponds to the cross-sectional view of representative cell types within the device (i) endothelium, ii) synovial gel, and iii) tendon) and demonstrates the spatial localization of FLS (orange), monocytes (red), and tendon fibroblasts (green) at Day 1 of culture. ECs and tendon resident monocyte-derived macrophages (MDM) are present but not labeled.

Article Snippet: FLS were lifted from flasks by first rinsing with HBSS (Gibco, 14175095) and applying 0.25% trypsin-EDTA for 3 min. FLS were encapsulated in a type I collagen hydrogel (Advanced Biomatrix, 5026) at a density of 100,000 cells ml -1 .

Techniques: Construct, Isolation, Functional Assay, Immunofluorescence, Derivative Assay, Labeling

Journal: bioRxiv

Article Title: A human synovial tendon-on-a-chip models key features of peritendinous adhesions and offers a new approach methodology for testing anti-fibrotic drugs

doi: 10.64898/2026.04.03.716316

Figure Lengend Snippet: a , An overview of the culture timeline to establish an FLS-EC co-culture. FLS are encapsulated in a type I collagen hydrogel and added to the basal side of the nanomembrane. After 24 h, ECs are added to the top vascular compartment and form a vascular barrier over 24 h. Media is replaced daily until Day 3 (D 3 ) at which point an inflammatory stimulus (10 ng ml⁻¹ TGF-β1) is added to the basal compartment. b, FLS-EC co-culture at Day 3 stained for VE-cadherin (green), cadherin-11 (yellow), and Hoechst (blue). Maximum intensity projection taken at 10x (left). i) z-slice focusing on the endothelial monolayer. ii) z-slice focusing near the bottom of the FLS gel, demonstrating robust VE-cadherin junctions in the ECs above the FLS gel and compartmentalization of cadherin-11 expression in the FLS gel. c, Synovial hydrogel stained for PRG4 (cyan) and F-actin (red) showing a z-slice in the FLS gel focusing just under the endothelium layer on the nanomembrane demonstrating robust PRG4 secretion from FLS closest to the endothelial barrier. d, Immunofluorescence images of collagen III (green) expression in the synovial gel, with Hoechst (blue) as nuclear counterstain. Density of collagen III+ cells upon stimulation with TGF-β1 on Day 6. e, Immunofluorescence images of nuclear Ki67 (yellow) expression in the synovial gel, with Hoechst (blue) as nuclear counterstain. The number of Ki67+ cells mm -3 significantly increased at Day 6 with TGF-β1 stimulation. f, Quantification of the total cell density in the synovial gel, showing comparable increases in cell counts at Day 6 regardless of TGF-β1 stimulation. One-way ANOVA with Tukey’s post-hoc test: n=3 devices per condition, *p<0.05, **p<0.01.

Article Snippet: FLS were lifted from flasks by first rinsing with HBSS (Gibco, 14175095) and applying 0.25% trypsin-EDTA for 3 min. FLS were encapsulated in a type I collagen hydrogel (Advanced Biomatrix, 5026) at a density of 100,000 cells ml -1 .

Techniques: Co-Culture Assay, Staining, Expressing, Immunofluorescence