Spatial correlation of native and engineered cartilage components at micron resolution

Tissue engineering (TE) approaches are being widely investigated for repair of focal defects in articular cartilage. However, the amount and/or type of extracellular matrix (ECM) produced in engineered constructs does not always correlate with the resultant mechanical properties. This could be related to the specifics of ECM distribution throughout the construct. Here, we present data on the amount and distribution of the primary components of native and engineered cartilage (i.e., collagen, proteoglycan (PG), and water) using Fourier transform infrared imaging spectroscopy (FT-IRIS). These data permit visualization of matrix and water at 25 μm resolution throughout the tissues, and subsequent colocalization of these components using image processing methods. Native and engineered cartilage were cryosectioned at 80 μm for evaluation by FT-IRIS in the mid-infrared (MIR) and near-infrared (NIR) regions. PG distribution correlated strongly with water in native and engineered cartilage, supporting the binding of water to PG in both tissues. In addition, NIR-derived matrix peaks correlated significantly with MIR-derived collagen peaks, confirming the interpretation that these absorbances arise primarily from collagen and not PG. The combined use of MIR and NIR permits assessment of ECM and water spatial distribution at the micron level, which may aid in improved development of TE techniques.