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Abstract
Purpose: Mechanical stimulation, a crucial factor for maintaining the cartilaginous phenotype and promoting the chondrogenesis, has been widely used in autologous chondrocyte transplantation. This study was designed to investigate a novel concept of mechanical refractory period of chondrocytes after dynamic hydrostatic pressure (dHP).
Materials and methods: dHP protocols (0.1 Hz, 2 MPa) were applied. The variation in type II collagen (Col II) expression induced by each dHP unit was measured. The dynamic remodeling of F-actin during the mechanical protocols was observed morphologically and mechanically by laser confocal microscopy and optical magnetic twisting cytometry (OMTC), respectively. About 20 ng/ml VEGF was used to stabilize the F-actin and restrain the mechanical refractory period.
Results: Compared with the remarkable increase of Col II (16-fold) induced by the initial dHP unit, the chondrocytes entered a mechanical refractory period and the second unit hardly elevated Col II expression (only 2.9-fold). This refractory period recovered partially within 2 h. The uniform, parallel, and coarse fibers of F-actin before dHP became thin, sparse, and disordered, and the cell stiffness decreased concomitantly. The variations in both the morphology and the mechanical property of F-actin were highly synchronous to the mechanical refractory period and recovered in a time-dependent manner. VEGF postponed the appearance of this refractory period and maintained the high expression of Col II by VEGF/p38/MAPKAPK-2/LIMK/cofilin pathway.
Conclusion: A mechanical refractory period of chondrocytes has been discovered and defined in this study. The F-actin depolymerization is the putative mechanism, and this refractory period can be postponed by VEGF-induced F-actin stabilization.
Declaration of interest
The authors report that they have no conflicts of interest. The authors here alone are responsible for the content and writing of the paper. This study was supported by the following funds: The National Natural Science Foundation of China (No. 81171461), Science Foundation of Hunan Provincial Science and Technology Department (No. 2012WK3001), and The Natural Science Foundation of Hunan Province (No. 13JJ2013).