The influence of radiation-induced collagen chain fragmentation, crosslinking, and sequential irradiation on the high-cycle fatigue life of human cortical bone.

ABSTRACT

Both high-cycle fatigue life and fatigue crack propagation resistance of human cortical bone allograft are radiation dose-dependent between 0 and 25 kGy such that higher doses exhibit progressively shorter lifetimes. Recently, we have shown that collagen chain fragmentation and stable crosslink accumulation may contribute to the radiation dose-dependent loss in fatigue crack propagation resistance of human cortical bone. To our knowledge, the influence of these mechanisms on high-cycle fatigue life of cortical bone have not been established. Sequential irradiation has also been shown to mitigate the loss of fatigue life of tendons, however, whether this mitigates losses in fatigue life of cortical bone has not been explored. Our objectives were to evaluate the influence of radiation-induced collagen chain fragmentation and crosslinking on the high-cycle fatigue life of cortical bone in the dose range of 0-15 kGy, and to evaluate the capability of sequential irradiation at 15 kGy to mitigate the loss of high-cycle fatigue life and radiation-induced collagen damage. High-cycle fatigue life specimens from four male donor femoral pairs were divided into 5 treatment groups (0 kGy, 5 kGy, 10 kGy, 15 kGy, and 15 kGy sequentially irradiated) and subjected to high-cycle fatigue life testing with a custom rotating-bending apparatus at a cyclic stress of 35 MPa. Following fatigue testing, collagen was isolated from fatigue specimens, and collagen chain fragmentation and crosslink accumulation were quantified using SDS-PAGE and a fluorometric assay, respectively. Both collagen chain fragmentation (p = 0.006) and non-enzymatic crosslinking (p < 0.001) influenced high-cycle fatigue life, which decreased with increasing radiation dose from 0 to 15 kGy (p = 0.016). Sequential irradiation at 15 kGy did not offer any mitigation in high-cycle fatigue life (p = 0.93), collagen chain fragmentation (p = 0.99), or non-enzymatic crosslinking (p ≥ 0.10) compared to a single radiation dose of 15 kGy. Taken together with our previous findings on the influence of collagen damage on fatigue crack propagation resistance, collagen chain fragmentation and crosslink accumulation both contribute to radiation-induced losses in notched and unnotched fatigue life of cortical bone. To maximize the functional lifetime of radiation sterilized structural cortical bone allografts, pathways other than sequential radiation should be explored to mitigate collagen matrix damage.