Journal: bioRxiv

Article Title: Macrophage migration is differentially regulated by distinct ECM components

doi: 10.1101/2023.04.27.538597

Figure Lengend Snippet: (A) Representative images of fixed macrophages stained with Phalloidin 488 on 10 ìg/mL Poly-L-Lysine (P-L-L), Collagen (Col), Fibronectin (FN), Laminin (Lam), or Vitronectin (VN). Scale bar = 200 microns in each image. (B) Quantification of spread cell area in microns squared (**** = p<0.0001, FN n=967 cells, Lam n=846 cells pooled from 3 independent experiments). Means and standard error of the mean for each experiment are represented with black symbols, all data points are plotted and each experimental run is color-coded. Statistical analysis was Kruskal-Wallis with Dunn multiple comparisons test. (C) Quantification of cellular elongation (A.U.). Greater elongation corresponds to a higher value in this analysis. (**** = p<0.0001, FN n=967 cells, Lam n=846 cells pooled from the same 3 independent experiments used to generate data in Figure 1B). Means and standard error of the mean for each experiment are represented with black symbols, all data points are plotted and each experimental run is color-coded. Statistical analysis was Kruskal-Wallis with Dunn multiple comparisons test. (D) Total fluorescence of phalloidin staining (i.e. actin filaments) on each ECM. (**** = p<0.0001, FN n=967 cells, Lam n=846 cells pooled from the same 3 independent experiments used to generate data in Figure 1B and 1C). Means and standard error of the mean for each experiment are represented with black symbols, all data points are plotted and each experimental run is color-coded. Statistical analysis was Kruskal-Wallis with Dunn multiple comparisons test. (E) Representative tracks of macrophages migrating randomly on 10 ìg/mL fibronectin or laminin over a 16 hour time period, generated by the TrackMate FIJI plugin. Scale bar = 200 microns. (F) Velocity (cell speed) in microns per minute and (G) Persistence (d/T) of randomly migrating macrophages on 10 ìg/mL of the indicated ECM (**** = p<0.0001, *** = p=0.0001, P-L-L n=1264 tracks, Col n=959 tracks, FN n=1004 tracks, Lam n=1559 tracks, VN n=773 tracks pooled from 2 experiments). Means and standard error of the mean for each experiment are represented with black symbols, all data points are plotted and each experimental run is colorcoded. Statistical analysis was Kruskal-Wallis with Dunn multiple comparisons test.

Article Snippet: Poly-L-lysine (Sigma-Aldrich, #P8920); Rat tail collagen, type I (ThermoFisher, #A1048301); Fibronectin, human plasma (ThermoFisher, #33016015); Laminin 111, mouse (ThermoFisher, #23017015); Vitronectin, human plasma (Sigma-Aldrich, #5051); Rhodamine-conjugated fibronectin (Cytoskeleton Inc., FNR01-A); HiLite 488-labeled laminin (Cytoskeleton Inc., LMN02-A)

Techniques: Staining, Fluorescence, Generated

Journal: Materials

Article Title: A Vitronectin-Derived Bioactive Peptide Improves Bone Healing Capacity of SLA Titanium Surfaces

doi: 10.3390/ma12203400

Figure Lengend Snippet: Cell attachment, spreading, and migration of osteoblast-like HOS cells seeded on culture plates treated with vitronectin and synthetic peptides. ( A ) Photographs of osteoblast-like HOS cells adhering (upper panel) and spreading (lower panel) to culture plates treated with 1% bovine serum albumin (BSA), vitronectin (0.26 μg/cm 2 ), scrambled peptide (SP), and VnP-16 peptide (10.5 μg/cm 2 ). Bar = 100 μm. ( B , C ) Cell attachment ( B ) and spreading ( C ) to immobilized synthetic peptides. HOS cells were allowed to adhere to peptide-treated plates for 1 h ( B ) or 3 h ( C ) in serum-free medium. ( D ) Migration of osteoblast-like HOS cells induced by vitronectin and synthetic peptides. HOS cells were seeded into the upper chambers of transwell filters coated with vitronectin (0.26 μg/cm 2 ), SP, or VnP-16 (10.5 μg/cm 2 ) and were incubated for 24 h. ND, not detected. ( E ) The viabilities of osteoblast-like HOS cells treated with VnP-16 for 24 or 48 h. ** p < 0.01 vs. the SP-treated control group. Data in ( B – E ) ( n = 4) represent the mean ± SD.

Article Snippet: Human plasma vitronectin was obtained from Millipore (Bedford, MA, USA).

Techniques: Cell Attachment Assay, Migration, Incubation

Journal: Materials

Article Title: A Vitronectin-Derived Bioactive Peptide Improves Bone Healing Capacity of SLA Titanium Surfaces

doi: 10.3390/ma12203400

Figure Lengend Snippet: Cell attachment and spreading of osteoblast-like MG-63 cells seeded on culture plates treated with vitronectin and synthetic peptides. ( A ) Photographs of osteoblast-like MG-63 cells adhering (upper panel) and spreading (lower panel) to culture plates treated with 1% bovine serum albumin (BSA), vitronectin (0.26 μg/cm 2 ), scrambled peptide (SP), and VnP-16 peptide (10.5 μg/cm 2 ). Bar = 100 μm. ( B – C ) Cell attachment ( B ) and spreading ( C ) to immobilized synthetic peptides. MG-63 cells were allowed to adhere to peptide-treated plates for 1 h ( B ) or 3 h ( C ) in serum-free medium. ( D ) The viabilities of osteoblast-like MG-63 cells treated with VnP-16 for 24 or 48 h. ** p < 0.01 vs. the SP-treated control group. Data in ( B – D ) ( n = 4) represent the mean ± SD.

Article Snippet: Human plasma vitronectin was obtained from Millipore (Bedford, MA, USA).

Techniques: Cell Attachment Assay