From fluctuations to stability: In-Situ chondrocyte response to cyclic compressive loading.

Chondrocytes, the sole cellular components in articular cartilage, are mechanosensitive and undergo significant morphological and volumetric changes in response to mechanical loading. These changes activate ion channels, initiating cellular mechanotransduction processes crucial for maintaining cartilage health. Dynamic loading has been shown to elicit anabolic responses that preserve cartilage integrity, while prolonged mechanical unloading leads to atrophy. However, the intricacies of how chondrocytes respond to dynamic loading remain poorly understood, largely due to technical limitations in capturing real-time cellular responses during loading cycles. This study aimed to advance our understanding of chondrocyte behavior during dynamic cyclic compression loading through high-speed imaging techniques. We developed a protocol to capture changes in chondrocyte volume, shape, and surface area at critical moments of maximal and minimal tissue stress during cyclic loading. Our findings revealed that chondrocyte volume fluctuated cyclically during the first 20 loading cycles, increasing by up to 4 % during load application and decreasing by as much as 8 % during unloading. These volume fluctuations stabilized over time, returning to baseline levels after approximately 100 cycles. Volume changes over time translate to shape change, causing similar oscillatory pattern in cell width and depth strains but not height strain, which remained relatively constant throughout the loading protocol. Changes in surface area mirrored the volume changes but were less pronounced (< 2 % increase), suggesting a protective mechanism against cell membrane rupture. This research offers valuable insights into the dynamic behavior of chondrocytes during cyclic loading, highlighting the importance of considering dynamic environments in cellular biomechanics studies.