In vivo gene detection is huge challenge in research and disease diagnosis. Authors developed a novel delivery method (nanopieces) to apply molecular beacon (MB) technology in in vivo Matrix metalloproteinase-13 (MMP-13) mRNA detection. MMP-13 is identified as biomarker for Osteoarthritis (OA), which is one of the most common causes of disability. This research will provide great tool for early diagnosis of OA. We designed MB for mouse MMP-13, GAPDH, and scramble sequence, and validated through primary mouse chondrocytes in vitro delivery. We performed Destabilization of the medial meniscus (DMM) on mouse animal OA model, and use Nanopieces delivering MB to monitor gene expression. In fig. 1 Red color of GAPDH MB illustrated the successful and efficient delivery via Nanopieces. MMP-13 MB only showed its color in stimulated cells, which illustrated the specificity of design. In fig. 2, MMP-13 MB displayed strong fluorescence in DMM leg compared to sham leg; scramble MB in another channel, which is co-delivered to eliminate the affection of non-specific degradation, showed weak signal. After calculation, MMP-13 MB real signal was about 4 times stronger in DMM leg than sham leg. (Fig. 2C) In this study, Nanopieces successfully delivered MB in vitro and in vivo. In vitro, MMP-13 MB was validated the effectivity in cells. In vivo MMP-13 MB was successfully delivered into live mouse cartilage in DMM mouse model. Significantly stronger signal of DMM leg to sham leg was observed, which provided an easy discrimination of inflammatory and non-inflammatory leg. Therefore we developed novel MMP-13 detection method, which is suitable for early, sensitive and long-lasting in vivo arthritis diagnosis.

In vivo quantification of MMP-13 using molecular beacon to realize early, sensitive, and long-lasting arthritis diagnosis

250th National Meeting (2015)

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