Journal: Biomaterials

Article Title: Host type 2 immune response to xenogeneic serum components impairs biomaterial-directed osteo-regenerative therapies

doi: 10.1016/j.biomaterials.2022.121601

Figure Lengend Snippet: Day 7 hydrogel explants were digested and a Luminex bead-based multiplex ELISA was run to analyze the local cytokine milieu. (a) Heatmap with two-way hierarchical clustering using Ward’s method with Euclidian distances shows distinct clustering of FBS-containing dorsal hydrogels based on local cytokine secretion. Scale bar represents data that has been log transformed, unit variance scaled, and mean centered. (b) Principal component analysis of cytokines within dorsal hydrogels. Principal component 1 and principal component 2 are displayed on the X- and Y-axes and explain 36.1% and 20.8% of the variance, respectively. 95% prediction ellipses are also plotted. (c) Cytokines with significantly different local concentrations, as well as key type 1 and type 2 immune response cytokines with no significant differences in concentrations (n=6 samples per group). Red, dashed line indicates the limit of detection of the Luminex MAGPIX System. A one-way ANOVA was used to detect statistical differences followed by Tukey’s multiple comparisons test for IL-4, Eotaxin, IL-9, IL-25, and IL-33. For all other cytokines, a Kruskal-Wallis test was used followed by Dunn’s multiple comparison test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Samples were analyzed using a Milliplex Mouse Cytokine/Chemokine Magnetic Bead Panel – Premixed 32 Plex (Millipore, MCYTOMAG-70K) or a custom R&D Luminex kit containing the analytes IFNγ, IL-13, IL-17A, IL-25, and IL-33 (R&D, LXSAMSM-05) on a MAGPIX System (Luminex Corp.) according to manufacturer instructions.

Techniques: Luminex, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Transformation Assay, Comparison