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)-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)-and IL-4 release was enhanced in the 3D system (relative to controls), there was a suppression of transforming growth factor (TGF)-production; effects on IL-10 secretion were variable. The results here suggested that collagen-chitosan scaffolds could provide a proinflammatory 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.

Macrophage interactions with polylactic acid and chitosan scaffolds lead to improved recruitment of human mesenchymal stem/stromal cells: a comprehensive study with different immune cells

“…Indeed, cell culture in a threedimensional spatial arrangement has been reported to affect a range of cellular functions, including cell proliferative index and gene expression, compared with two-dimensional substrates [27,28]. Moreover, similar observations were found between three-dimensional matrices with distinct properties [29].…”

Follistatin‐like protein 1 contributes to dendritic cell and T‐lymphocyte activation in nasopharyngeal carcinoma patients by altering nuclear factor κb and Jun N‐terminal kinase expression

“…This investigation found that DC immunity in NPC patients was augmented by FSTL1. Similarly, TNF‐α has been shown to promote the development of immature DCs into mature DCs . Therefore, the mechanism by which FSTL1 expedites the immune ability of DCs should be elucidated from this perspective.…”

“…Similarly, TNF-α has been shown to promote the development of immature DCs into mature DCs. 54,55 Therefore, the mechanism by which FSTL1 expedites the immune ability of DCs should be elucidated from this perspective. Above all, it is important to investigate the mechanism by which FSTL1 affects DCs to provide a basis for new antitumor immunotherapies.…”

Chitosan/collagen blends with inorganic and organic additive—A review

“…[69] The addition of collagen to chitosan increases pores diameter and porosity of the scaffold. [71] [71] …”