jbma37326-sup-0001-FigureS1.docxWord 2007 document , 337.7 KB

Figure S1 Example of a typical Western blot as used for evaluation of M1 and M2 macrophage marker protein expression, here depending on chitosan concentration (Cp), see Figure 4. Western blots were cut in half, and the upper part reacted with mouse anti-iNOS, or goat anti-CD206 antibodies, respectively; the lower part with mouse anti-actin antibodies and in case of M2 macrophages with rabbit anti-Arg-1 antibodies. Approximate molecular weights are indicated to the right (the two Western blots were not cut in exactly the same way and the extreme lower part was cut off). At least three independent experiments (from monocyte culture to Western blotting) were performed for statistical evaluation. Western blots were scanned, and band intensities analyzed using the LiCor ImageStudio Lite application. Actin band intensities served as reference for total protein content of a sample.

jbma37326-sup-0002-FigureS2.docxWord 2007 document , 726.3 KB

Figure S2 Effect of chitosan Cp and DA on CD206 expression of macrophages. CD206 levels expressed by M2 macrophages are lower on tissue culture plastic (TCP) than on chitosan substrate, but no significant differences are observed between chitosan hydrogels differing in their DA or Cp.

jbma37326-sup-0003-FigureS3.docxWord 2007 document , 4.3 MB

Figure S3 M1 andM2 macrophages seeded on poly-l-lysine, and 2% and 15% DA chitosan-FPHS, 36 h after addition of polarizing factors. Note that cell densities are about equal on the different substrates (see the DAPI only staining, bottom), but on 15% DA chitosan macrophages show aggregation, accompanied by reduced cell spreading, indicative of a lower substrate adhesivity (combined phalloidin/DAPI staining, top). Scale bar, 100 μm.

jbma37326-sup-0004-TableS1.docxWord 2007 document , 107.3 KB

Table S1 Statistical evaluation of relative macrophage marker protein expression levels iNOS, Arg-1 and CD206 depending on chitosan concentration (Cp, left column), and degree of acetylation (DA, right column), as shown in Figure 4, and Figure S2. One-way ANOVA with Tukey's multiple comparisons post-hoc test, n = 3–5 independent experiments.

jbma37326-sup-0005-TableS2.docxWord 2007 document , 63.9 KB

Table S2 Statistical evaluation of relative marker protein expression levels of macrophages grown on tissue culture plastic (TCP), with chito-oligosaccharides (COS) added to the medium, on chitosan alone, or on chitosan with chito-oligosaccharides (COS) added (C + C), as in Figure 7. One-way ANOVA with Tukey's multiple comparisons post-hoc test, n = 3–5 independent experiments.