Nthesis after 48 h of loading. However, it has been shown that

Nthesis after 48 h of loading. However, it has been shown that changes in the biosynthesis may not be related solely to changes in mRNA expression [55]. While aggrecan and collagen II mRNA were up-regulated during the initial 0.5 h of static compression and decreased during the following 4?4 h, the synthesis of aggrecan and collagen protein decreased more rapidly already after 0.5 h [55]. However, none of the reviewed studies investigated collagen II or proteoglycans at both the mRNA and the protein level. Furthermore, the mRNA level alone does not give information about how the extracellular matrix is adapted in response to the loading. The Basmisanil web secretion and assembly of protein into the extracellular space is essential to change the mechanical properties of the tissue. Therefore, when investigating extracellular matrix proteins, like collagen II or proteoglycans, further investigations should include not only mRNA analysis but especially a detailed analysis of the extracellular amount and spatial distribution pattern of the proteins. Hence, it would be of interest to distinguish between soluble protein that is released into the supernatant and protein that is embedded into extracellular structures.PLOS ONE | DOI:10.1371/journal.pone.0119816 March 30,13 /Cyclic Tensile Strain and Chondrocyte MetabolismSuperficial Zone Protein. The superficial zone protein contributes to the lubrication function of the surface layer or articular cartilage which is essential for nearly frictionless gliding of the articulating joint partners under motion [56]. CTS of 7 upregulated mRNA levels of superficial zone protein after 12, 24 and 48 h compared to levels before loading [45] and compared to unloaded cells [57]. Higher strains (21 ) elevated mRNA levels after 12 h loading compared to levels before loading [45] and compared to unloaded cells [57]. Nevertheless, it decreased under control levels after 48 h of loading. Accordingly, immunoblot analysis revealed that superficial zone protein levels increased under 7 strain and decreased under 21 strain [57]. The results suggest that moderate loading supports lubrication and low-friction-motion by increasing expression of superficial zone protein in chondrocytes. Mechanical overloading, however, inhibits the expression and synthesis and thereby provokes cartilage degradation under motion since lubrication function is disturbed. Fibronectin. Fibronectin connects collagen fibers and other ECM proteins [58]. It is linked to the cell membrane through integrins and might transmit forces from the ECM to the chondrocyte [59]. CTS at 7 , 0.33 Hz and 0.5 Hz, for 4, 12 and 24 h increased the fibronectin mRNA levels in comparison to non-loaded chondrocytes [33,60]. This suggests that tissue adaptation in response to moderate CTS also FT011 structure comprises the production of molecules that are involved in matrix-cell connection and mechanical signal transmission, like fibronectin. To our knowledge, other non-collagenous matrix proteins have not yet been investigated in response to CTS in monolayer. However, it has been shown in three-dimensional agarose constructs that for example the cartilage oligomeric matrix protein (COMP) was increased in response to cyclic tension in chondrocytes [61]. Further investigation is needed to understand the complex interplay of mechanical signals and matrix adaptation. Information about the effect of two-dimensional CTS on non-collagenous proteins like the adhesive glycoproteins thrombospondin or cho.Nthesis after 48 h of loading. However, it has been shown that changes in the biosynthesis may not be related solely to changes in mRNA expression [55]. While aggrecan and collagen II mRNA were up-regulated during the initial 0.5 h of static compression and decreased during the following 4?4 h, the synthesis of aggrecan and collagen protein decreased more rapidly already after 0.5 h [55]. However, none of the reviewed studies investigated collagen II or proteoglycans at both the mRNA and the protein level. Furthermore, the mRNA level alone does not give information about how the extracellular matrix is adapted in response to the loading. The secretion and assembly of protein into the extracellular space is essential to change the mechanical properties of the tissue. Therefore, when investigating extracellular matrix proteins, like collagen II or proteoglycans, further investigations should include not only mRNA analysis but especially a detailed analysis of the extracellular amount and spatial distribution pattern of the proteins. Hence, it would be of interest to distinguish between soluble protein that is released into the supernatant and protein that is embedded into extracellular structures.PLOS ONE | DOI:10.1371/journal.pone.0119816 March 30,13 /Cyclic Tensile Strain and Chondrocyte MetabolismSuperficial Zone Protein. The superficial zone protein contributes to the lubrication function of the surface layer or articular cartilage which is essential for nearly frictionless gliding of the articulating joint partners under motion [56]. CTS of 7 upregulated mRNA levels of superficial zone protein after 12, 24 and 48 h compared to levels before loading [45] and compared to unloaded cells [57]. Higher strains (21 ) elevated mRNA levels after 12 h loading compared to levels before loading [45] and compared to unloaded cells [57]. Nevertheless, it decreased under control levels after 48 h of loading. Accordingly, immunoblot analysis revealed that superficial zone protein levels increased under 7 strain and decreased under 21 strain [57]. The results suggest that moderate loading supports lubrication and low-friction-motion by increasing expression of superficial zone protein in chondrocytes. Mechanical overloading, however, inhibits the expression and synthesis and thereby provokes cartilage degradation under motion since lubrication function is disturbed. Fibronectin. Fibronectin connects collagen fibers and other ECM proteins [58]. It is linked to the cell membrane through integrins and might transmit forces from the ECM to the chondrocyte [59]. CTS at 7 , 0.33 Hz and 0.5 Hz, for 4, 12 and 24 h increased the fibronectin mRNA levels in comparison to non-loaded chondrocytes [33,60]. This suggests that tissue adaptation in response to moderate CTS also comprises the production of molecules that are involved in matrix-cell connection and mechanical signal transmission, like fibronectin. To our knowledge, other non-collagenous matrix proteins have not yet been investigated in response to CTS in monolayer. However, it has been shown in three-dimensional agarose constructs that for example the cartilage oligomeric matrix protein (COMP) was increased in response to cyclic tension in chondrocytes [61]. Further investigation is needed to understand the complex interplay of mechanical signals and matrix adaptation. Information about the effect of two-dimensional CTS on non-collagenous proteins like the adhesive glycoproteins thrombospondin or cho.

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