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Objective To study the low-intensity pulsed ultrasound on rat PSCs proliferation and secretion of extracellular matrix , and to explore the possible mechanism . Isolated neonatal rat neonatal rat the PSCs cells using immunomagnetic beads sorting isolated purified rat PSCs , the use of specific cell surface markers FGFR-3 immunohistochemical identification . Take well-grown third-generation cells with trypsin digestion , cells were resuspended in serum-containing medium and counted under a microscope , to of 1.6X10 << / sup > sup> 3 / ml were seeded in 96-well plates , divided into four groups ( blank control group , 5 minutes / day group , 12 min / day group , 20 minutes / day group ) in each of four wells . At 37 ℃, saturated humidity , 5 ? 2 incubator cultured 24h, low-intensity pulsed ultrasound intervention the ( ultrasonic strength 100mW/cm 2 When the cells adherent frequency of 1MHz, pulse interval is 20%). Stimulation when the coupling agent evenly on the ultrasonic probe , the probe is placed in the bottom of the culture plates were stimulated cells stimulated cultured for 24h . Stimulus after the first day , the third and the fifth day with CCK-8 assay cell proliferation . The other to 2X10 5 a / ml were seeded cells in 35mm diameter cell culture dishes grouping with before each group of three wells . Intervene in the same way as before , at intervals of 30 minutes after stimulation extraction of total RNA by RT-PCR method to detect collagen type II , Aggrecan, TGF-β 1 and Sox9 gene expression . Results LIPU a promoting effect the SD rats PCSCs proliferation , proliferation more obvious in the first day only 20 minutes / day stimulation group difference was statistically significant compared with the control group ; continuous stimulation in three days and five days after the experimental and control groups showed significant differences , but the differences between the various experimental groups was not significant . LIPU of type II collagen , of SD rats PCSCs 's Aggrecan , TGF - β 1 < / sub> and expression of Sox9 gene mRAN have obvious role in promoting more multi - mRAN expression more stimulate days to 20 minutes / day stimulation group increased the most obvious. Conclusion LIPU promote the SD rats PCSCs proliferation ability , and can promote the expression of the extracellular matrix mRAN , this effect may be by the TGF -beta 1 and Sox9 gene mediated 's .
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