Zahm J M, Kaplan H, Hérard A L, Doriot F, Pierrot D, Somelette P, Puchelle E
INSERM U314, IFR53, CHU Maison Blanche, Reims, France.
Cell Motil Cytoskeleton. 1997;37(1):33-43. doi: 10.1002/(SICI)1097-0169(1997)37:1<33::AID-CM4>3.0.CO;2-I.
The respiratory epithelium is frequently injured by inhaled toxic agents or by micro-organisms. The epithelial wound repair represents a crucial process by which surface respiratory cells maintain the epithelial barrier integrity. The repair process involves both cell migration and proliferation, but as yet, the kinetic of these two mechanisms has not been extensively studied. Using an in vitro model of human respiratory epithelium wound repair, proliferative cell immunofluorescent staining and a computer-assisted technique allowing the tracking of living cells, we studied the cell proliferation and migration during the wound repair process. Respiratory epithelial cells were dissociated from human nasal polyps and cultured on a collagen I matrix. At confluency, a chemical wound was made on the culture. We observed that the cell mitotic activity peaked at 48 h after wounding (23% of the cells) and mainly concerned the cells located 160 to 400 microns from the wound edge. The migration speed was highest (35 to 45 microns/h) for the spreading cells at the wound edge and progressively decreased for the cells more and more distant from the wound edge. The temporal analysis of the cell migration speed during the wound repair showed that it was almost constant during the first 3 days of the repair mechanism and thereafter dropped down until the wound closure was completed (after 4 days). We also observed that over a 1-hour period, the intra-individual and interindividual variation of the cell migration speed was 43% and 37%, respectively. These results demonstrate that cell proliferation and cell migration during respiratory epithelial wound repair are differently expressed with regard to the cell location within the repairing area.
呼吸道上皮经常受到吸入的有毒物质或微生物的损伤。上皮伤口修复是一个关键过程,通过这个过程,表面呼吸道细胞维持上皮屏障的完整性。修复过程涉及细胞迁移和增殖,但到目前为止,这两种机制的动力学尚未得到广泛研究。我们使用人类呼吸道上皮伤口修复的体外模型、增殖细胞免疫荧光染色以及一种允许追踪活细胞的计算机辅助技术,研究了伤口修复过程中的细胞增殖和迁移。呼吸道上皮细胞从人鼻息肉中分离出来,培养在I型胶原基质上。细胞汇合后,在培养物上制造化学伤口。我们观察到,细胞有丝分裂活性在受伤后48小时达到峰值(23%的细胞),主要涉及位于距伤口边缘160至400微米处的细胞。伤口边缘正在铺展的细胞迁移速度最高(35至45微米/小时),而离伤口边缘越来越远的细胞迁移速度逐渐降低。对伤口修复过程中细胞迁移速度的时间分析表明,在修复机制的前3天,迁移速度几乎恒定,此后下降,直到伤口完全闭合(4天后)。我们还观察到,在1小时内,细胞迁移速度的个体内和个体间变异分别为43%和37%。这些结果表明,呼吸道上皮伤口修复过程中的细胞增殖和细胞迁移在修复区域内的细胞位置方面表现不同。