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Impaired neutrophil directional chemotactic accuracy in chronic periodontitis patients.

Roberts HM, Ling MR, Insall R, Kalna G, Spengler J, Grant MM, Chapple IL - J. Clin. Periodontol. (2015)

Bottom Line: Circular statistics were utilized for the analysis of cell movement.However, velocity and accuracy were normalized for the weak chemoattractant CXCL8 while they remained significantly reduced for fMLP.Dysfunctional neutrophil chemotaxis may predispose patients with periodontitis to their disease by increasing tissue transit times, thus exacerbating neutrophil-mediated collateral host tissue damage.

View Article: PubMed Central - PubMed

Affiliation: Periodontal Research Group and MRC Centre for Immune Regulation, University of Birmingham, Birmingham, UK.

No MeSH data available.


Related in: MedlinePlus

Photograph of the Insall chamber. Large square illustrates position of the coverslip with adhered neutrophils. Arrows show the application of the chemoattractant. Small red rectangle shows the area visualized by video microscopy.
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fig02: Photograph of the Insall chamber. Large square illustrates position of the coverslip with adhered neutrophils. Arrows show the application of the chemoattractant. Small red rectangle shows the area visualized by video microscopy.

Mentions: The Insall chamber was used to visualize chemotaxis (Muinonen-Martin et al. 2010). For each sample, isolated neutrophils (400 μl in RPMI, final density 1 × 106/ml) were added to acid washed (0.2 M HCl), dried and blocked (7.5%, BSA 400 μl, Sigma) coverslips (22 mm, VWR International), which were then incubated at room temperature (approximately 23°C) for 30 min. to allow the cells to adhere. The coverslip was then inverted and placed at the top of the chemotaxis chamber ensuring that the chemoattractant loading bays were exposed (Fig. 2). The desired chemoattractant (80 µl, fMLP (10 nM) or CXCL8 (used at 200 ng/ml after assessing a range of concentrations) or control (RPMI media) was injected into the chemoattractant channels. Cell movement was analysed using a Zeiss Primovert microscope (Carl Zeiss Imaging, Thornwood, NY, USA) and Images captured every 30 s for up to 40 frames per condition using a Q Imaging Retiga 2000R camera (Qimaging, Surry, Canada).


Impaired neutrophil directional chemotactic accuracy in chronic periodontitis patients.

Roberts HM, Ling MR, Insall R, Kalna G, Spengler J, Grant MM, Chapple IL - J. Clin. Periodontol. (2015)

Photograph of the Insall chamber. Large square illustrates position of the coverslip with adhered neutrophils. Arrows show the application of the chemoattractant. Small red rectangle shows the area visualized by video microscopy.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340045&req=5

fig02: Photograph of the Insall chamber. Large square illustrates position of the coverslip with adhered neutrophils. Arrows show the application of the chemoattractant. Small red rectangle shows the area visualized by video microscopy.
Mentions: The Insall chamber was used to visualize chemotaxis (Muinonen-Martin et al. 2010). For each sample, isolated neutrophils (400 μl in RPMI, final density 1 × 106/ml) were added to acid washed (0.2 M HCl), dried and blocked (7.5%, BSA 400 μl, Sigma) coverslips (22 mm, VWR International), which were then incubated at room temperature (approximately 23°C) for 30 min. to allow the cells to adhere. The coverslip was then inverted and placed at the top of the chemotaxis chamber ensuring that the chemoattractant loading bays were exposed (Fig. 2). The desired chemoattractant (80 µl, fMLP (10 nM) or CXCL8 (used at 200 ng/ml after assessing a range of concentrations) or control (RPMI media) was injected into the chemoattractant channels. Cell movement was analysed using a Zeiss Primovert microscope (Carl Zeiss Imaging, Thornwood, NY, USA) and Images captured every 30 s for up to 40 frames per condition using a Q Imaging Retiga 2000R camera (Qimaging, Surry, Canada).

Bottom Line: Circular statistics were utilized for the analysis of cell movement.However, velocity and accuracy were normalized for the weak chemoattractant CXCL8 while they remained significantly reduced for fMLP.Dysfunctional neutrophil chemotaxis may predispose patients with periodontitis to their disease by increasing tissue transit times, thus exacerbating neutrophil-mediated collateral host tissue damage.

View Article: PubMed Central - PubMed

Affiliation: Periodontal Research Group and MRC Centre for Immune Regulation, University of Birmingham, Birmingham, UK.

No MeSH data available.


Related in: MedlinePlus