Abstract
BackgroundCollagen cross-links contribute to the mechanical resilience of the intervertebral disc (IVD). UVA-light-activated riboflavin-induced collagen crosslinking (UVA-CXL) is a well-established and effective ophthalmological intervention that increases the mechanical rigidity of the collagen-rich corneal matrix in Keratoconus. This study explores the feasibility, safety and efficacy of translating this intervention in reinforcing the IVD.MethodsAnnulus fibrosus (AF) cells were isolated from bovine IVDs and treated with different combinations of riboflavin (RF) concentrations (0.05–8 mM) and UVA light intensities (0.3–4 mW/cm2). Metabolic activity (resazurin assay), cell viability (TUNEL assay), and gene expression of apoptosis regulators C-FOS and PT5 were assessed immediately and 24 hours after treatment. Biomechanical effects of UVA-CXL on IVDs were measured by indentation analysis of changes in the instantaneous modulus and by peel-force delamination strength analysis of the AF prior and after treatment.ResultsDifferent intensities of UVA did not impair the metabolic activity of AF cells. However, RF affected metabolic activity (p < 0.001). PT53 expression was similar in all RF conditions tested while C-FOS expression decreased 24 hours after treatment. Twenty-four hours after treatment, no apoptotic cells were observed in any condition tested. Biomechanical characterizations showed a significant increase in the annular peel strength of the UVA-CXL group, when compared to controls of UVA and RF alone (p < 0.05). UVA-CXL treated IVDs showed up to 152% higher (p < 0.001) instantaneous modulus values compared to the untreated control.ConclusionThis is the first study on UVA-CXL treatment of IVD. It induced significantly increased delamination strength and instantaneous modulus indentation values in intact IVD samples in a structure–function relationship. RF concentrations and UVA intensities utilized in ophthalmological clinical protocols were well tolerated by the AF cells. Our findings suggest that UVA-CXL may be a promising tool to reinforce the IVD matrix.
Highlights
Back pain is the most important single cause of disability worldwide, preventing patients from engaging in work and other everyday activities [1,2,3]
Different intensities of UVA did not impair the metabolic activity of annulus fibrosus (AF) cells
PT53 expression was similar in all RF conditions tested while C-FOS expression decreased 24 hours after treatment
Summary
Back pain is the most important single cause of disability worldwide, preventing patients from engaging in work and other everyday activities [1,2,3]. It is mostly associated with pathologies of the intervertebral disc (IVD). Progressive aggrecan degradation leads to a reduced water-binding capacity [5, 6]. These alterations cause debilitation of tissue strength and loss of mechanical properties, thereby contributing to biomechanical failure, disc herniation, instability and pain [8]. This study explores the feasibility, safety and efficacy of translating this intervention in reinforcing the IVD
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