Fig 1: Cumulative release studies in AEC-CM loaded HA-PEG and HA-PEG-HEP hydrogels. (A) Cumulative total protein release profile from HA-PEG and HA-PEG-HEP hydrogels in 1× PBS at 37 °C for 21 days and the magnified cumulative release profile during the initial 6 h. (B) Cumulative VEGF release profile from HA-PEG and HA-PEG-HEP hydrogels in 1× PBS at 37 °C for 21 days and the magnified cumulative release profile during the initial 6 h. (C) Cumulative AREG release profile from HA-PEG and HA-PEG-HEP hydrogels in 1× PBS at 37 °C for 21 days and the magnified cumulative release profile during the initial 6 h. Results are presented as mean ± SD (n ≥ 3), and the samples were analyzed with ordinary one-way ANOVA, followed by Turkey posthoc test, setting at the probabilities *p < 0.05, **p < 0.01, ***p< 0.001, ****p< 0.0001 comparing each HA-PEG and HA-PEG-HEP sample within the same time point.
Fig 2: Histograms representing (A) the FE (%) of CMINF total proteins absorbed on the 3D scaffolds subjected to three different functionalization methods: PA, HCl, and NaOH pretreatments; and (B) AREG quantification in CMINF (red line) or adsorbed on the 3D scaffold functionalized with PA or NaOH pre-activation. * Statistically significant between groups for p < 0.05.
Fig 3: Chemical characterization of 3D scaffolds before and after functionalization. Infrared spectra for (A) liquid (SF, SF+AREG, and CMINF), (B) PA, (C) HCl, and (D) NaOH-treated PLGA scaffolds, functionalized or not with SF+AREG and CMINF, and reported in the 650 and 4000 cm−1.
Fig 4: Experimental design. (A) Functionalization of 3D scaffolds with CMINF (3D+CMINF), production of released media (CMR) at different time points (6, 24, 48, 72 h, and 7 d), and quantification of AREG from the collected CMR. (B) Biological activity assessment of 3D+CMINF (direct effect) and each CMR collected at different time points: 6, 24, 48, 72 h, and 7 d (indirect effect) on activated PBMCs and Jurkat cells after 48 h of culture. Analysis of the 3D+CMINF influence on macrophage polarization was performed on total, scaffold-adherent, and supernatant fractions.
Fig 5: Kinetic release profile of AREG from 3D+CMINF and 3D+AREG at different time points. The red line represents the maximum concentration of AREG adsorbed on the surface of 3D scaffolds previously exposed to NaOH pre-treatment.
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