Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Measurements of total protein and growth factors released from hPLG and xPMC/hPLG. (A) Cumulative release percentage of total protein over a 4 day period, determined by BCA assay. (B) SDS-PAGE analysis results of the PBS supernatants collected over a 4 day period, stained with Coomassie blue. Cumulative release of (C) TGF-β1 and (D) PDGF-BB over a 3 day period. (E) Total protein contents found in the PBS supernatants of hPLG and hPLG combined with either PM or xCMC, assessed after a 2 day incubation. Data are expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01. Statistical significance in (C,D) was analyzed at the same time point.

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: BIA-KA, SDS Page, Staining, Incubation

Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Effect of xPMC/hPLG on the migration of periodontal fibroblasts. (A) Cells were seeded and exposed to chemokines from different gel spots for 6 h. The migratory cells surrounding the edge of gel spots of (i) hPPG, (ii) hPLG, (iii) xPMC/hPLG, (iv) xPMC/hPLG pretreated with IgG isotype control antibody (Ab), and (v) xPMC/hPLG pretreated with neutralizing antibody (nAb) against PDGF-BB were stained with crystal violet (A), and the cell density (cells/mm 2 ) was quantified from the number of cells that migrated close to each gel spot within a 500 μm radial distance from the gel edge (B). The results are expressed as mean ± SD ( n = 8). Different letters indicate significant differences ( p < 0.05).

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: Migration, Control, Staining

Journal: ACS Omega

Article Title: In Vitro Enhanced Performance of Human Platelet Lysate Gel Integrated with Mesoporous Silica Nanoparticle/Carboxymethyl Chitosan Composite Hydrogel: Structural Stability and Biological Activities for Chronic Wound Healing

doi: 10.1021/acsomega.5c13494

Figure Lengend Snippet: Schematic image of the proposed concentration-dependent biphasic effect of xPMC/hPLG on the chemotaxis and cell doubling of periodontal fibroblasts. Color gradient represents a concentration gradient from high (red) to low (blue) concentrations of hPL-derived mediators released from the xPMC/hPLG hydrogel. The hydrogel gave rise to a concentration gradient of a GF, i.e., PDGF-BB, declining with increasing distance away from the hydrogel. Fibroblasts in Area 2 were exposed to a low concentration of PDGF-BB sufficient to induce their migration toward the hydrogel. As the cells ascended the concentration gradient, the signal for chemotaxis was switched off, but the signal for cell division was switched on, thereby resulting in the presence of proliferating cells at Area 1, adjacent to the hydrogel.

Article Snippet: To unequivocally determine the role of PDGF-BB specifically as a primary GF mediating hPL-induced periodontal fibroblast migration, PDGF-BB activity was inhibited by preincubating xPMC/hPLG spots with 0.2 μg/mL of a PDGF-BB neutralizing antibody (AF-220-NA, R&D Systems) for 45 min. As a negative control, xPMC/hPLG spots were pretreated for 45 min with an isotype control goat IgG antibody (0.2 μg/mL; AB-108 C, R&D Systems).

Techniques: Concentration Assay, Chemotaxis Assay, Derivative Assay, Migration