RESUMO
Mechanical signals regulate a multitude of cell functions and ultimately govern fibrous tissue growth, maintenance and repair. Such mechanotransduction processes often involve modulation of intracellular calcium concentration ([Ca2+]i). However, most studies interrogate these responses in cells in simplified culture systems, thereby removing potentially important inputs from the native extracellular microenvironment. The objective of this study was to test the hypothesis that the intracellular calcium response of meniscus fibrochondrocytes (MFCs) is dependent on both the microenvironmental context in which this perturbation is applied and on the tensile deformation. Using a custom micro-mechanical tester mounted on a confocal microscope, intracellular calcium activity in MFCs in response to incremental tissue strains (0, 3, 6 and 9 %) was monitored in situ (i.e., in the native tissues) on MFC-seeded aligned scaffolds and MFC-seeded silicone membranes. The [Ca2+]i regulation by MFCs within the native meniscus tissue microenvironment was considerably different from [Ca2+]i regulation by MFCs on either aligned nanofibrous scaffolds or flat silicone membranes. Additionally, increasing levels of tensile deformation resulted in a greater number of responding cells, both in situ and in vitro, while having no effects on temporal characteristics of [Ca2+]i signalling. Collectively, these findings have significant implications for mechanobiology of load-bearing fibrous tissues and their responses to injury and degeneration. In addition, from a tissue engineering perspective, the findings establish cellular benchmarks for maturing engineered constructs, where native tissue-like calcium mechano-regulation may be an important outcome parameter to achieve mechanical functionality comparable to native tissue.