Effect of testing temperature on the nanostructural response of tendon to tensile mechanical overload.

Despite many in vitro mechanical experiments of tendon being conducted at room temperature, few assessments have been made to determine how the structural response of tendon to mechanical overload may vary with ambient temperature. We explored whether damage to the collagen nanostructure of tendon resulting from tensile rupture varies with temperature. Use of bovine tail tendons in combination with NaBH crosslink stabilization treatment allowed us to probe the mechanisms underlying the observed changes. Untreated tendons and NaBH-stabilized tendons were pulled to rupture at temperatures of 24, 37, and 55 °C. Of nine mechanical parameters measured from the resulting stress-strain curves, only yield stress differed between the tendons tested at 37 and 24 °C. When tested at 55 °C, untreated tendons showed large reductions in ultimate strength and toughness, while NaBH-stabilized tendons showed smaller reductions. Differential scanning calorimetry was used to assess damage to the collagen fibril nanostructure of tendons resulting from rupture, with samples from the ruptured tendons compared to samples from the same tendons removed prior to loading. While there was indication that overload-induced molecular packing disruption to collagen fibrils may be heightened at 37 °C, statistical increases in damage compared to that occurring at 24 °C were only seen when testing was conducted at 55 °C. The results show that the temperature sensitivity of tendon to ramp loading depends on crosslinking within the tissue. In poorly crosslinked tissues, collagen may be more susceptible to mechanical damage when tested at physiologic temperature compared to room temperature. For tendons with a high density of thermally stable crosslinks, such as the human Achilles or patellar tendons, testing at room temperature should produce comparable results to testing at physiologic temperature.