Effects of vibration and hyaluronic acid on activation of three‐dimensional cultured chondrocytes

Abstract

Objective. To investigate the effects of vibration (Vib) and hyaluronic acid (HA) on 3-dimensional cultured cartilage.Methods. Chondrocytes were obtained from metatarsophalangeal joints of freshly killed 6-month-old pigs. Twenty-four-well plates containing type I collagen sponge disks were used to culture samples. The frequency and the amplitude of the vibration of the well plate were 100 Hz and 0.5 nm, respectively. We produced 3-dimensional cartilage tissue using HA and vibration with collagen sponge as a carrier. Four different culture conditions were examined: a control HA؊Vib؊ group, an HA؊Vib؉ group, an HA؉Vib؊ group, and an HA؉Vib؉ group. Each group was cultured for 2 weeks. After culture days 3, 7, 10, and 14 (every 3.5 days), the levels of chondroitin 4-sulfate (C4S) and chondroitin 6-sulfate (C6S) isomers synthesized in each culture medium were measured. Histologic analysis, immunohistochemical analysis, and electron microscopic examination were performed.Results. Mean C4S and C6S synthesis had increased rapidly after 7 days of culture and continued to increase thereafter. There were significant differences among the 4 groups (P < 0.01). Synthesis of both C4S and C6S was most abundant in the HA؉Vib؉ group and the lowest in the HA؊Vib؊ group. After 1 and 2 weeks of culture, the chondrocytes had formed stratified structures on the collagen sponges in all groups, although the thickest structure was observed in the HA؉Vib؉ group and the thinnest in the HA؊Vib؊ group. Under immunofluorescence, the HA؉Vib؉ group exhibited the strongest chromatic features. Under electron microscopy, the chondrocytes in the HA؉Vib؉ group exhibited many long and slender prominences on their surface, and extracellular substance could be observed associated with the cells.Conclusion. Our results indicate that the combination of vibration and HA activates the production of proteoglycan in 3-dimensional cultured chondrocytes and stimulates MAPK and ␤-catenin. This suggests that some mechanoreceptors for vibration exist on the plasma membrane of chondrocytes and activate the intracellular signal transduction system.

The effect of bioreactor induced vibrational stimulation on extracellular matrix production from human derived fibroblasts

“…The evidence that does exist is for cells derived from tissue other than the vocal fold but does support the idea that vibrational stimuli can increase matrix synthesis. Takeuchi et al demonstrated that a high frequency (100 Hz) low displacement (0.5 nm) stimulus was capable of increasing chondrocyte matrix synthesis [28]. Specifically, they demonstrated that both chondroitin and collagen type II production was increased following vibrational stimulation.…”

Design and characterization of a dynamic vibrational culture system

“…The shear forces introduced to the cell membrane could cause changes in intracellular pressure. Further, the acceleration induced by vibration could be perceived by the cells (Takeuchi et al ., ). These factors combined lead to conformational changes of transmembrane proteins, such as mechanosensitive ion channels or integrin receptors (Wang et al ., ).…”

Separating Fluid Shear Stress from Acceleration during Vibrations In Vitro: Identification of Mechanical Signals Modulating the Cellular Response

“…25,31,32,41,47,50 While these studies have been defining the biochemical response of the cell, the physical mechanism by which the signal is sensed and transduced is typically neglected. Primarily based on in vivo investigations, direct and indirect mechanisms have been suggested including out-of-phase acceleration of the cell nucleus 38 or fluid shear.…”

“…Exploiting the opportunities of cell culture systems to investigate the underlying mechanisms, distinct cell types including osteoblasts, 2 osteocytes, 31 myoblasts, 50 chondrocytes, 47 or progenitor cells 45 have been shown to respond to vibrations in vitro. While providing important data on the biologic response of cells to vibrations, the identification of the specific component(s) of the vibratory signal that modulates the response requires the quantification of the cellular mechanical environment.…”