Effects of high-intensity cyclical hydrostatic pressures on the ultrastructure and type II collagen expression of chondrocytes of knee joint in human
WANG Xin
BAI Qian
XU Kui
QIU Xiu-chun
FAN Qing-yu
MA Bao-an
Abstract：Objective To investigate the effects of high-intensity cyclic hydrostatic pressures on the ultrastructure and type II collagen contents of chondrocytes of the knee joint cultured in vitro in human. Methods The normal chondrocytes of human knee joint were isolated and cultured in vitro. The 3rd generation of chondrocytes were treated with high-intensity cyclical hydrostatic pressures ( 10.0 mPa ) by the multifunctional thermostatic high-insensitive hydrostatic pressure loading device for 2 h per day lasting for 5 days. Toluidine blue staining and immunohistochemical staining of type II collagen were employed to identify the chondrocytes. Cell morphology was observed by light microscopy. The ultrastructure of chondrocytes were observed by transmission electron microscopy ( TEM ). Type II collagen immunohistochemical staining and semi-quantitative analysis were performed to measure contents and expression of type II collagen of chondrocytes in 2 groups. Results Compared with the control group, cell morphology changed from irregular polygon into long spindle, membrane and cytoplasm retracted, the number of cells was reduced significantly and grew sparsely in the 10.0 mPa group. TEM showed that various apoptosis indications such as uneven distribution and margination of chromosomes in the 10.0 mPa group. Collagen type II immunohistochemistry and semi-quantitative analysis showed that the percent of stained area of control group and 10.0 mPa group was ( 61.64 ± 6.19 ) % and ( 43.76 ± 5.61 ) %, compared with the control group, the stained area and extent of chondrocytes were significantly decreased in the 10.0 mPa group ( P < 0.05 ). Conclusions The high-intensity pressure over human physiological range can be regarded as a mechanical injury to result in cell apoptosis, morphology and intracellular ultrastructural changes of human chondrocytes, and also decrease protein expression of human chondrocytes. These data provide the experimental basis for studying the relationship between stress injury and pathogenesis of osteoarthritis.
Keywords：ChondrocytesPressureUltrastructureTransmission electron microscopyCollagen type II chondrocytes pressureMicroscopyelectrontransmissionCollagen type II
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 920-924 )