sdf-1α solution pbs  (PeproTech)

Journal: Acta biomaterialia

Article Title: A Liposome/Gelatin Methacrylate Nanocomposite Hydrogel System for Delivery of Stromal Cell-Derived Factor-1 and Stimulation of Cell Migration

doi: 10.1016/j.actbio.2020.03.015

Figure Lengend Snippet: Schematic of the overall delivery system, which is composed of liposomal SDF-1α (lipoSDF) loaded in a UV-crosslinkable gelatin methacrylate (GelMA) hydrogel derived from Type B gelatin. Anionic liposomes are formed by mixing DSPC, DSPG, and cholesterol at a 65:10:25 molar ratio.

Article Snippet: The 1 mL mixture was placed on a rotovap overnight (Büchi R-100) at 40 °C and 100 mBar to remove the chloroform and generate a thin lipid film, which was subsequently rehydrated with 1 mL SDF-1α solution in PBS (12.5 μg/mL, Peprotech).

Techniques: Derivative Assay

Journal: Acta biomaterialia

Article Title: A Liposome/Gelatin Methacrylate Nanocomposite Hydrogel System for Delivery of Stromal Cell-Derived Factor-1 and Stimulation of Cell Migration

doi: 10.1016/j.actbio.2020.03.015

Figure Lengend Snippet: (A) Cumulative SDF-1α released from liposomes over 1 week is characterized by burst release kinetics (n = 4, mean ± STD). (B) SEM imaging of GelMA + lipoSDF shows small particles of less than 500 nm dispersed throughout the surface of a hydrogel fiber (Scale bar: 5 μm). (C) Cumulative percentage of unencapsulated SDF-1α or (E) lipoSDF released from GelMA over 1 week, normalized to the amount of initial detectable protein (n = 4, mean ± STD). One-way ANOVAs with Tukey’s post-hoc comparisons were conducted between groups at each time point. *p < 0.0001, **p = 0.001, ***p < 0.01). (D) Release exponent (n) and r2 values derived from the release of unencapsulated SDF-1α from GelMA hydrogels, fit to the Korsmeyer-Peppas model. (F) Shape parameter (β) and r2 values derived from release of lipoSDF, fit to the Weibull release model.

Article Snippet: The 1 mL mixture was placed on a rotovap overnight (Büchi R-100) at 40 °C and 100 mBar to remove the chloroform and generate a thin lipid film, which was subsequently rehydrated with 1 mL SDF-1α solution in PBS (12.5 μg/mL, Peprotech).

Techniques: Imaging, Derivative Assay

Journal: Acta biomaterialia

Article Title: A Liposome/Gelatin Methacrylate Nanocomposite Hydrogel System for Delivery of Stromal Cell-Derived Factor-1 and Stimulation of Cell Migration

doi: 10.1016/j.actbio.2020.03.015

Figure Lengend Snippet: (A) Representative images and (B) quantitative cell counts from Hoechst-stained MSCs in a region of interest (ROI) that have migrated through a transwell membrane in response to the respective chemotactic factor: PBS, 80 ng/mL of free or liposomal SDF-1α (Scale bar: 100 μm. n = 3, mean ± STD. One-way ANOVAs with Tukey’s post-hoc comparisons, *p < 0.01). (C) Comparison of MSC migration in response to media conditioned over 1 week by PBS, 80 ng/mL SDF-1α, or either 5 μg/mL of free or liposomal SDF-1α in 10% GelMA. Two sets of one-way ANOVAs with Tukey’s post-hoc comparisons were conducted between groups (groups being compared are denoted by either upper- or lower-case letters). Groups that do not share a letter are statistically different (p < 0.05).

Article Snippet: The 1 mL mixture was placed on a rotovap overnight (Büchi R-100) at 40 °C and 100 mBar to remove the chloroform and generate a thin lipid film, which was subsequently rehydrated with 1 mL SDF-1α solution in PBS (12.5 μg/mL, Peprotech).

Techniques: Staining, Migration

Journal: Acta biomaterialia

Article Title: A Liposome/Gelatin Methacrylate Nanocomposite Hydrogel System for Delivery of Stromal Cell-Derived Factor-1 and Stimulation of Cell Migration

doi: 10.1016/j.actbio.2020.03.015

Figure Lengend Snippet: (A) Representative Western blots of phosphorylated AKT and RPS6 compared to their respective total protein controls in MSCs exposed to PBS, 80 ng/mL free SDF-1α, or 5 μg/mL of either free or liposomal SDF-1α in GelMA. (B) Densitometry analysis of phosphorylated AKT and (C) RPS6. (n = 3, mean ± STD. Two sets of one-way ANOVAs with Tukey’s post-hoc comparisons were conducted between groups. *p < 0.05 and **p < 0.01)

Article Snippet: The 1 mL mixture was placed on a rotovap overnight (Büchi R-100) at 40 °C and 100 mBar to remove the chloroform and generate a thin lipid film, which was subsequently rehydrated with 1 mL SDF-1α solution in PBS (12.5 μg/mL, Peprotech).

Techniques: Western Blot