The ciliary beating of upper respiratory tract cells cultured on cover glasses was studied by using differential interference microscope equipped with high speed video. By culturing the cells on collagen coated cover glasses, objectives with higher magnifications could be used. With this system we could evaluate not only ciliary beat frequency, but also amplitude, wave form, orientation and synchrony of ciliary beating. Also some structural anomalies such as compound cilia and tide cilia bundles could be recognized. Ciliary beat frequency measured from 1,026 ciliated cells was 20.6 +/- 4.7 Hz (mean +/- SD). The orientation of ciliary beat directions was random and the mean standard deviation for measured angles was 73.0 degrees +/- 28.9 degrees (mean SD +/- SD). When the ciliary beat frequency was 20 Hz, the time used for effective phase was 0.022 +/- 0.002 s (mean +/- SD), and 0.028 +/- 0.004 s (mean +/- SD) for the recovery phase of beat. This system is advantageous for studying ciliary function because all parts can be studied simultaneously with higher magnification, and the effects of chemical physical mediators can be studied without disturbing effects of the autonomic nervous system or secretory cells. Also, the same cells could be observed before and after challenge with test medication and thus evaluated more accurately.
To study the detailed effects of exogenous adenosine triphosphate (ATP) on ciliary function, we used the differential interference microscope equipped with high speed video and evaluated ciliated cells from the human sinus mucosa in monolayer culture. With this system it was possible to evaluate all parts of ciliary motility with a minimum of interference from the mucous membrane, secretory cells and the autonomic nervous system. The best direct ciliostimulative effect of exogenous ATP on ciliary beat frequency (CBF) and ciliary beat amplitude (CBA) was observed at concentrations of ATP ranging from 10(-5) M to 10(-3) M. Exogenous ATP appeared to normalize the slightly damaged ciliary motility in groups with an initial CBF of 14 Hz or higher. When the CBF was less than 14 Hz, ATP produced an increase in CBF greater than 40%, an increase in CBA greater than 30%, but these did not reach the normal level in 5 min. The biggest increases: 59.9% in CBF and 40.7% in CBA were seen in the group with an initial CBF less than 9 Hz. In the cells with a low initial CBF and unsynchronized motility exogenous ATP increased the synchrony of ciliary movement together with an increase in CBF and CBA.
The degeneration of ciliary beat of human respiratory cells was studied in monolayer cell cultures by using a differential interference microscope equipped with a high speed video system. This method for studying ciliary beat in cell cultures on collagen-coated cover glasses is quite advantageous, because it allows for detailed study of all parts of ciliary function and not just ciliary beat frequency (CBF). In the present study both CBF and ciliary beat amplitude (CBA) were found to decrease continuously from the 1st day after plating but the wave form of ciliary beat did not change. Cultures with high cell density provided better preservation of normal ciliary beat for a longer period. In contrast, ciliary beat degenerated quickly in cultures with low cell density. CBF and CBA in cell cultures less than 5 days after plating were always high, supporting use of these cultures for studies of normal ciliary motility.
Soft palate stiffening operations--often with CO2 or contact Nd:YAG lasers--have been used recently to treat patients with pronounced snoring. Differences in soft palate scars formed after CO2 and contact Nd:YAG laser incisions were studied in an animal model to determine which of these lasers produces a more rigid scar and consequently might be more effective for a soft palate stiffening operation. Six mongrel dogs were placed in each laser group, after which either the CO2 or Nd:YAG laser was used to make a 1.5-cm-long soft palate incision. The scars produced were then excised and studied histologically 4 weeks postoperatively. Special attention was paid to collagen density and elastin fibers in the scar tissue. After contact Nd:YAG laser incisions scar tissue was found to contain densely aligned collagen fibers and practically no elastin fibers. CO2 laser incisions produced significantly different scars: irregularly arranged collagen fibers with some elastin fibers also inside the scar tissue. Differences in the shapes of the scars and wound contractions were also found that were only seen after contact laser incisions. These findings suggest that the contact Nd:YAG laser might be more effective for soft palate stiffening operations.
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