Journal: Biomaterials Science

Article Title: Modular noncovalent functionalization of electrospun piezoelectric scaffolds with bioactive nanocarriers

doi: 10.1039/d5bm01563d

Figure Lengend Snippet: Electrostatic liposome adsorption onto PVDF-TrFE scaffolds: (A) fluorescence microscopy of Cy5 labeled liposomes on scaffolds coated in deionized water, HEPES, and HEPES-NaCl (top to bottom) at low NP concentration (0.5 mg mL −1 ) or high NP concentration (2 mg mL −1 ) on poled scaffolds, or high NP concentration on unpoled scaffolds (left to right); scale bar = 100 µm and is applied to all groups. Images are collected at the same laser power and are contrast-matched using Fiji-ImageJ. (B) Image-based quantification of liposome adsorption onto scaffolds. (C) Energy-dispersive X-ray spectroscopy measurements of nitrogen levels on unpoled scaffolds coated with 2 mg mL −1 of liposomes in HEPES-NaCl, (D) piezoelectric coefficient, d 33 , on unpoled scaffolds, uncoated or coated with 2 mg mL −1 of liposomes in HEPES-NaCl, pre- and post-ethanol wash. All values are represented as mean ± SEM and pair-wise statistics are performed using Student's paired t -tests. Multiple comparisons following two-way ANOVA for image-based fluorescence quantification and d 33 measurements are corrected by Tukey's multiple comparisons test.

Article Snippet: The thin film was rehydrated at the desired concentration in loading buffer in a bath sonicator at 60 °C for 30 min. For fluorescent liposomes only, 1,2-distearoyl- sn-glycero -3-phosphoethanolamine- N -(Cyanine 5) fluorescent lipid (18:0 Cy5 PE) (Avanti Polar Lipids #810345) was added at 0.01 mol% during sonication.

Techniques: Adsorption, Fluorescence, Microscopy, Labeling, Liposomes, Concentration Assay, Spectroscopy

Journal: Biomaterials Science

Article Title: Modular noncovalent functionalization of electrospun piezoelectric scaffolds with bioactive nanocarriers

doi: 10.1039/d5bm01563d

Figure Lengend Snippet: Liposome coated scaffolds are suitable substrates for cells and enable cytokine presentation: (A) live HEK293 cell counts were measured from multiple time points on unpoled scaffolds with and without cationic liposome coating. Data is presented as mean ± SEM from 3 independent wells at each time point. Statistical significance was assessed by two-way ANOVA with Sidak's multiple comparisons test, comparing coated to uncoated scaffold at each time point. P values reflect differences between groups at individual time points, and all error bars are present on graph although some are too small to see. (B) Unpoled scaffolds were coated with fluorescent Cy5-labeled liposomes and imaged with confocal microscopy. Jurkat T cells were stained with calcein, cultured on Cy5 liposome-coated scaffolds, and imaged at 60× resolution 3 days after plating. (C) Jurkat T cells were fixed on day 4 and stained intracellularly with ActinProbe555 and DAPI. Arrows indicate punctate actin (yellow) or liposome uptake (magenta). Scale bars are 50 µm or 10 µm. (D) Activating the JAK-STAT Pathway with IL-15 Liposomes on Scaffolds: (i) unpoled scaffolds were coated with 2 mg mL −1 of IL-15-tethered liposomes in optimized HEPES-NaCl buffer conditions and SEAP is produced in response to IL-15 recognition. (ii) IL-15 reporter cells were plated on unpoled scaffolds, and absorbance was read daily, correlating to the downstream SEAP production from activation of the JAK-STAT pathway with n = 3 samples per group. Data is presented as relative fold change from uncoated scaffold control, with mean ± SD. Significance is conducted with an Ordinary Two-Way Anova with multiple comparisons using Šídák's multiple comparisons test.

Article Snippet: The thin film was rehydrated at the desired concentration in loading buffer in a bath sonicator at 60 °C for 30 min. For fluorescent liposomes only, 1,2-distearoyl- sn-glycero -3-phosphoethanolamine- N -(Cyanine 5) fluorescent lipid (18:0 Cy5 PE) (Avanti Polar Lipids #810345) was added at 0.01 mol% during sonication.

Techniques: Labeling, Liposomes, Confocal Microscopy, Staining, Cell Culture, Produced, Activation Assay, Control