During aging, wear, and tear of intervertebral discs, human discs undergo a series of morphological and biochemical changes. Degradation of extracellular matrix proteins, e.g., collagen, arises as an important contributor and accelerator in this process. Existing methods to detect collagen degradation at the tissue level include histology and immunohistochemistry. Unfortunately, most of these methods only depict overall collagen content without the ability to specifically discern degraded collagen and to assess the severity of degeneration. To fill this technological gap, we developed a robust and simple approach to detect and assess early disc degeneration with a collagen hybridizing peptide (CHP) that hybridizes with the flawed triple helix structure in degraded collagen. Intriguingly, the CHP signal in mouse lumbar discs exhibited a linear incremental pattern with age. This finding was corroborated with histological analysis based on established methods. When comparing this analysis, a positive linear correlation was found between CHP fluorescence intensity and the histological score with a regression value of r(2) = 0.9478. In degenerative mouse discs elicited by pro-inflammatory stimuli (IL-1 beta and LPS) ex vivo, the newly developed approach empowered prediction of the severity of disc degeneration. We further demonstrated higher CHP signals in a degenerative human disc tissue when compared to a normal sample. These findings also resonated with histological analysis. This approach lays a solid foundation for specific detection and assessment of intervertebral disc degeneration at the molecular level and will promote development of future disc regeneration strategies.
[1]Univ Virginia, Dept Orthopaed Surg, 135 Hosp Dr,Cobb Hall, Charlottesville, VA 22908 USA;
[2]Huazhong Univ Sci & Technol, Tongji Hosp, Dept Orthopaed, Tongji Med Coll, 1095 Jiefang Ave, Wuhan 430030, Hubei, Peoples R China;
[3]Univ Utah, Dept Biomed Engn, 201 Presidents Circle, Salt Lake City, UT 84112 USA;
[4]Fudan Univ, Huashan Hosp, Dept Orthopaed, 12 Wulumuqi Middle Rd, Shanghai 200040, Peoples R China;
[5]Univ Virginia, Dept Biomed Engn, 415 Lane Rd, Charlottesville, VA 22904 USA
(8.0+极速模式)