Fibroblast orientation to stretch begins within three hours

Most connective tissue cells align in response to stretch. Previous studies have shown these responses occur within 12-14 h of initiation of stretch, but do not identify the time at which this orientation occurs, nor whether the orientation continues after cessation of stretch. To ascertain the earliest times at which fibroblast orientation occurs, we cultured primary human fibroblasts on deformable culture dishes and stretched (1 Hz, 8% uniaxial strain) them for up to 24 h. We photographed the cells at 0.5, 1-6, 8, 10, 12, 14, 16, and 24 h. Similarly cells were photographed at 1-3, or 4 h after cessation of stretch for stretch durations of 1, 2, and 3 h. Orientation of cells were ascertained by an interactive computer program. The fibroblasts began to orient by 2-3 h and orientation appeared nearly complete by 24 h. Cultures stretched for 2 or 3 h continued to exhibit greater degrees of orientation (compared to controls) for 2 or 3 h respectively after cessation of stretch. We conclude fibroblasts begin to orient within 3 h of initiation of stretch, and that they continue to orient for several hours after cessation of stretch.