PURPOSE: The current study was designed to determine if a polymorphonuclear leukocyte (PMN) chemoattractant is derived from alkali-degraded whole cornea and to establish a range for its molecular weight. METHODS: We utilized a collagen gel-visual chemotactic assay to quantify the directional movement of PMN exposed to alkali-degraded corneas (30 min or 24 hr). In this experiment, the sample to be tested for chemotactic activity passed through a 14,000 molecular weight cutoff membrane into a collagen gel in which resting neutrophils were suspended in a random fashion. Cell movement was videotaped and subsequently tracked by digitizing the centroid of the cells at 30-sec intervals. Computer analysis of these tracks illustrated many behavioral characteristics, including directional movement. RESULTS: Alkali-degraded whole bovine corneas produced a chemotactic response in neutrophils within 5 min. Dilution of the 24-hr sample to 1:7 yielded a significant chemotactic response. The chemotactic response of both the 30-min and 24-hr samples followed a dose-response curve. CONCLUSIONS: This agent may be one of the inflammatory mediators that trigger the early neutrophil response after an alkali-injury to the eye.
PURPOSE: The current study was designed to determine if a polymorphonuclear leukocyte (PMN) chemoattractant is derived from alkali-degraded whole cornea and to establish a range for its molecular weight. METHODS: We utilized a collagen gel-visual chemotactic assay to quantify the directional movement of PMN exposed to alkali-degraded corneas (30 min or 24 hr). In this experiment, the sample to be tested for chemotactic activity passed through a 14,000 molecular weight cutoff membrane into a collagen gel in which resting neutrophils were suspended in a random fashion. Cell movement was videotaped and subsequently tracked by digitizing the centroid of the cells at 30-sec intervals. Computer analysis of these tracks illustrated many behavioral characteristics, including directional movement. RESULTS: Alkali-degraded whole bovine corneas produced a chemotactic response in neutrophils within 5 min. Dilution of the 24-hr sample to 1:7 yielded a significant chemotactic response. The chemotactic response of both the 30-min and 24-hr samples followed a dose-response curve. CONCLUSIONS: This agent may be one of the inflammatory mediators that trigger the early neutrophil response after an alkali-injury to the eye.
Authors: Saskia A Overbeek; Paul A J Henricks; Anja I Srienc; Pim J Koelink; Petra de Kruijf; Herman D Lim; Martine J Smit; Guido J R Zaman; Johan Garssen; Frans P Nijkamp; Aletta D Kraneveld; Gert Folkerts Journal: Eur J Pharmacol Date: 2011-03-31 Impact factor: 4.432