title: Effects of 3-dimensional culture conditions (collagen-chitosan nano-scaffolds) on maturation of dendritic cells and their capacity to interact with T-lymphocytes.
abstract: In the body, there is a natural three-dimensional (3D) microenvironment in which immune cells, including dendritic cells (DC), play their functions. This study evaluated the impact of using collagen-chitosan 3D nano-scaffolds in comparisons to routine 2D culture plates on DC phenotype and functions. Bone marrow-derived DC were cultured on scaffolds and plates and then stimulated with lipopolysaccharide (LPS) or chitosan-based nanoparticles (NP) for 24 h. Thereafter, DC viability, expression of maturation markers and levels of cytokines secretion were evaluated. In another set of studies, the DC were co-cultured with allogenic T-lymphocytes in both the 2D and 3D systems and effects on DC-induction of T-lymphocyte proliferation and cytokine release were analyzed. The results indicated that CD40, CD86 and MHC II marker expression and interleukin (IL)-12, IL-6 and tumor necrosis factor (TNF)-α secretion by DC were enhanced in 3D cultures in comparison to by cells maintained in the 2D states. The data also showed that DNA/chitosan NP activated DC more than LPS in the 3D system. T-Lymphocyte proliferation was induced to a greater extent by DNA/NP-treated DC when both cell types were maintained on the scaffolds. Interestingly, while DC induction of T-lymphocyte interferon (IFN)-γ and IL-4 release was enhanced in the 3D system (relative to controls), there was a suppression of transforming growth factor (TGF)-β production; effects on IL-10 secretion were variable. The results here suggested that collagen-chitosan scaffolds could provide a pro-inflammatory and activator environment to perform studies to analyze effects of exogenous agents on the induction of DC maturation, NP uptake and/or cytokines release, as well as for the ability of these cells to potentially interact with other immune system cells in vitro.
authors.affiliation: a Department of Immunology, Faculty of Medical Sciences , Tarbiat Modares University , Tehran , Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Daneshmandi
authors.affiliation: a Department of Immunology, Faculty of Medical Sciences , Tarbiat Modares University , Tehran , Iran.
authors.forename: Shaghayegh Pishkhan
authors.initials: SP
authors.lastname: Dibazar
authors.affiliation: a Department of Immunology, Faculty of Medical Sciences , Tarbiat Modares University , Tehran , Iran.
authors.forename: Shirin
authors.initials: S
authors.lastname: Fateh
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D048271:Chitosan / Q000737:chemistry*; D003094:Collagen / Q000737:chemistry*; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D003094:Collagen; D048271:Chitosan
keywords: Collagen-chitosan scaffold; T cells; cytokine; dendritic cells; maturation markers