Raman spectroscopic probe provides optical biomarkers of cartilage composition predictive of tissue function

The diagnosis of early osteoarthritis when therapeutic interventions may be most effective at reversing cartilage degeneration presents a clinical challenge. We describe a Raman arthroscopic probe and spectral analysis that measures biomarkers reflective of the content of predominant cartilage extracellular matrix (ECM) constituents-glycosaminoglycans (GAG), collagen, water-essential to cartilage function. We compare the capability of Raman-probe-derived biomarkers to predict functional properties of cartilage to quantitative MRI and histopathology assessments.

Osteochondral blocks were sectioned from 6 bovine femoral condyles with no macroscopic injury (n=62 blocks) and 6 condyles with a focal chondral lesion (n=32 blocks), but no macroscopic degeneration of surrounding cartilage (n=34 blocks). Blocks from 10 human knees were further analyzed (age 27-75; n=235 blocks). Using a custom arthroscopic Raman spectroscopy probe, spectra of chondral layers were measured and subjected to multivariate linear decomposition to extract ECM biomarker scores, reflecting the contribution of each ECM constituent to the spectra. Blocks were further analyzed for elastic modulus, T2/T2* MRI relaxation times, and OARSI scores.

For bovine tissues, Raman biomarkers revealed depleted GAG and cartilage softening peripheral to the lesion despite no macroscopic degeneration. Raman biomarkers accounted for 78% of GAG content variation and 71% of modulus variation. For human tissues, Raman biomarkers accounted for 71% of modulus variation. Raman biomarkers accounted for a greater variation of modulus (71%-72%) than OARSI (12-54%), T2* (15%-27%), or T2 (25%-30%).

These data support the application of Raman-probe-derived biomarkers for molecular assessment of key ECM constituents that define cartilage properties in health and disease.