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9, the Group G1 (Control), G2 (Toxic group), G3 (treatment with conventional 5-FU formulation), G4 (treatment with 1% 5-FU loaded chitosan nanogel), and G5 (treatment with 2% 5-FU loaded chitosan nanogel)

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Sections were then rinsed in PBS and incubated with the appropriate biotinylated secondary antibody at 1/1000 dilution for 1 h at room temperature

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The tissues were cut into pieces and digested with type I collagenase (1 mg/ml, Gibco, New York, USA) for 0.51 h at 37 C