In this Letter, we report a multiplexed polarized hypodermic Raman needle probe for the biostructural analysis of articular cartilage. Using a custom-developed needle probe with a sapphire ball lens, we measure polarized Raman spectra of cartilage. By imaging two polarizations simultaneously on the charge-coupled device (CCD) and binning them separately, we capture both biochemical and structural tissue information in real time. Here, we demonstrate that polarized Raman spectroscopy can distinguish between different collagen fibril alignment orientations in a cartilage explant model system, supporting its capacity for diagnosing the hallmark collagen alignment changes occurring in the early stages of osteoarthritis (OA). Accordingly, this work shows that needle-based polarized Raman spectroscopy has great potential for the monitoring and diagnosis of early OA.