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Villi-crypt in vitro model. Resazurin reduction assay performed (a) over the Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds in the presence and absence of gelatin to aid cell attachment ( n = 3 scaffolds for each condition). (b) Resazurin reduction assay performed for long-term cultures in the absence of gelatin (two independent experiments, n = 5 scaffolds for each condition of each experiment). Data are expressed as mean ± standard deviation. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (c) Phalloidin (green) and DAPI (blue) fluorescence microscopy <t>of</t> <t>Caco-2</t> cells grown for 21 days on the two compositions tested at two different magnifications. (c i ) Top and (c ii ) section view of the scaffolds. Scale bars represent 500 μm and 100 μm. Objectives are 4 × and 10 × , respectively. (d) Confocal images of Villi-(R)-Crypt scaffold cultured with the Caco-2 cells. (d i ) Schematic of the different confocal planes acquired. (d ii ) and (d iii ) details at two different magnifications of villi sections, scaffold baselines and crypt base; displaying the homogeneous covering of the scaffold and junctions formed between cells ( V = villi, C = crypt). Scale bars represent 200 μm (d ii , 10 × objective) and 50 μm (d iii , 20 × objective). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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Villi-crypt in vitro model. Resazurin reduction assay performed (a) over the Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds in the presence and absence of gelatin to aid cell attachment ( n = 3 scaffolds for each condition). (b) Resazurin reduction assay performed for long-term cultures in the absence of gelatin (two independent experiments, n = 5 scaffolds for each condition of each experiment). Data are expressed as mean ± standard deviation. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (c) Phalloidin (green) and DAPI (blue) fluorescence microscopy <t>of</t> <t>Caco-2</t> cells grown for 21 days on the two compositions tested at two different magnifications. (c i ) Top and (c ii ) section view of the scaffolds. Scale bars represent 500 μm and 100 μm. Objectives are 4 × and 10 × , respectively. (d) Confocal images of Villi-(R)-Crypt scaffold cultured with the Caco-2 cells. (d i ) Schematic of the different confocal planes acquired. (d ii ) and (d iii ) details at two different magnifications of villi sections, scaffold baselines and crypt base; displaying the homogeneous covering of the scaffold and junctions formed between cells ( V = villi, C = crypt). Scale bars represent 200 μm (d ii , 10 × objective) and 50 μm (d iii , 20 × objective). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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(A)CCK-8 assay demonstrating that the material alleviates oxidative stress and enhances cell survival. (B)Quantitative analysis of scratch healing rate. (C) Scratch wound healing assay showing cell proliferation capability. (D, E) EdU staining and PI staining assays <t>of</t> <t>Caco-2</t> cells following various treatments. (F, G) Quantitative analysis of EdU-positive cells, TUNEL-positive cells, and scratch closure rate. (H–J) mRNA expression levels of tight-junction proteins (Claudin-1, Occludin and ZO-1) in Caco-2 cells measured by quantitative qRT-PCR. (K) Heatmap of differentially expressed genes from RNA-seq. (L) PCA plot showing global transcriptome shifts. (M) Volcano plot of DEGs between H 2 O 2 and Co 3 O 4 @MMT groups. (N) KEGG enrichment analyses of DEGs. (O) Protein expression levels of HIF-1α, VEGFA and GLUT1 in Caco-2 cells. All statistical analyses were performed by comparing the H 2 O 2 group with other groups (n = 3), with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ns indicating no significant difference.
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Villi-crypt in vitro model. Resazurin reduction assay performed (a) over the Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds in the presence and absence of gelatin to aid cell attachment ( n = 3 scaffolds for each condition). (b) Resazurin reduction assay performed for long-term cultures in the absence of gelatin (two independent experiments, n = 5 scaffolds for each condition of each experiment). Data are expressed as mean ± standard deviation. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (c) Phalloidin (green) and DAPI (blue) fluorescence microscopy of Caco-2 cells grown for 21 days on the two compositions tested at two different magnifications. (c i ) Top and (c ii ) section view of the scaffolds. Scale bars represent 500 μm and 100 μm. Objectives are 4 × and 10 × , respectively. (d) Confocal images of Villi-(R)-Crypt scaffold cultured with the Caco-2 cells. (d i ) Schematic of the different confocal planes acquired. (d ii ) and (d iii ) details at two different magnifications of villi sections, scaffold baselines and crypt base; displaying the homogeneous covering of the scaffold and junctions formed between cells ( V = villi, C = crypt). Scale bars represent 200 μm (d ii , 10 × objective) and 50 μm (d iii , 20 × objective). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: Villi-crypt in vitro model. Resazurin reduction assay performed (a) over the Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds in the presence and absence of gelatin to aid cell attachment ( n = 3 scaffolds for each condition). (b) Resazurin reduction assay performed for long-term cultures in the absence of gelatin (two independent experiments, n = 5 scaffolds for each condition of each experiment). Data are expressed as mean ± standard deviation. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (c) Phalloidin (green) and DAPI (blue) fluorescence microscopy of Caco-2 cells grown for 21 days on the two compositions tested at two different magnifications. (c i ) Top and (c ii ) section view of the scaffolds. Scale bars represent 500 μm and 100 μm. Objectives are 4 × and 10 × , respectively. (d) Confocal images of Villi-(R)-Crypt scaffold cultured with the Caco-2 cells. (d i ) Schematic of the different confocal planes acquired. (d ii ) and (d iii ) details at two different magnifications of villi sections, scaffold baselines and crypt base; displaying the homogeneous covering of the scaffold and junctions formed between cells ( V = villi, C = crypt). Scale bars represent 200 μm (d ii , 10 × objective) and 50 μm (d iii , 20 × objective). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: In Vitro, Cell Attachment Assay, Standard Deviation, Fluorescence, Microscopy, Cell Culture

(a) Fold change in gene expression with respect to undifferentiated cells and (b) quantification of produced proteins. Caco-2 cells were cultured for 14 and 21 days on the two compositions tested ( n = 3 scaffolds for each condition). Data are expressed as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01.

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: (a) Fold change in gene expression with respect to undifferentiated cells and (b) quantification of produced proteins. Caco-2 cells were cultured for 14 and 21 days on the two compositions tested ( n = 3 scaffolds for each condition). Data are expressed as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: Gene Expression, Produced, Cell Culture, Standard Deviation

Venn diagram showing the distribution of proteins identified in Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds cultured with Caco-2 cells for the period of 14 and 21 days. The core proteins are shared across all conditions, while the exclusive proteins represent those uniquely identified in specific conditions. The 10 most abundant proteins are highlighted.

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: Venn diagram showing the distribution of proteins identified in Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds cultured with Caco-2 cells for the period of 14 and 21 days. The core proteins are shared across all conditions, while the exclusive proteins represent those uniquely identified in specific conditions. The 10 most abundant proteins are highlighted.

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: Cell Culture

(A)CCK-8 assay demonstrating that the material alleviates oxidative stress and enhances cell survival. (B)Quantitative analysis of scratch healing rate. (C) Scratch wound healing assay showing cell proliferation capability. (D, E) EdU staining and PI staining assays of Caco-2 cells following various treatments. (F, G) Quantitative analysis of EdU-positive cells, TUNEL-positive cells, and scratch closure rate. (H–J) mRNA expression levels of tight-junction proteins (Claudin-1, Occludin and ZO-1) in Caco-2 cells measured by quantitative qRT-PCR. (K) Heatmap of differentially expressed genes from RNA-seq. (L) PCA plot showing global transcriptome shifts. (M) Volcano plot of DEGs between H 2 O 2 and Co 3 O 4 @MMT groups. (N) KEGG enrichment analyses of DEGs. (O) Protein expression levels of HIF-1α, VEGFA and GLUT1 in Caco-2 cells. All statistical analyses were performed by comparing the H 2 O 2 group with other groups (n = 3), with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ns indicating no significant difference.

Journal: Materials Today Bio

Article Title: An orally deliverable Co 3 O 4 @MMT nanozyme platform for inflammatory bowel disease via ROS scavenging, barrier repair, and mucosal homeostasis regulation

doi: 10.1016/j.mtbio.2026.103310

Figure Lengend Snippet: (A)CCK-8 assay demonstrating that the material alleviates oxidative stress and enhances cell survival. (B)Quantitative analysis of scratch healing rate. (C) Scratch wound healing assay showing cell proliferation capability. (D, E) EdU staining and PI staining assays of Caco-2 cells following various treatments. (F, G) Quantitative analysis of EdU-positive cells, TUNEL-positive cells, and scratch closure rate. (H–J) mRNA expression levels of tight-junction proteins (Claudin-1, Occludin and ZO-1) in Caco-2 cells measured by quantitative qRT-PCR. (K) Heatmap of differentially expressed genes from RNA-seq. (L) PCA plot showing global transcriptome shifts. (M) Volcano plot of DEGs between H 2 O 2 and Co 3 O 4 @MMT groups. (N) KEGG enrichment analyses of DEGs. (O) Protein expression levels of HIF-1α, VEGFA and GLUT1 in Caco-2 cells. All statistical analyses were performed by comparing the H 2 O 2 group with other groups (n = 3), with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ns indicating no significant difference.

Article Snippet: The RAW264.7 mouse macrophages and human Caco-2 cell lines were both provided by the ATCC (USA).

Techniques: CCK-8 Assay, Wound Healing Assay, Staining, TUNEL Assay, Expressing, Quantitative RT-PCR, RNA Sequencing