Despite the substantial burden posed by osteoarthritis
(OA) globally,
difficult challenges remain in achieving early OA diagnosis and adopting
effective disease-modifying treatments. In this study, we use a biomolecular
approach to address these limitations by creating an inherently theranostic
molecular beacon whose imaging and therapeutic capabilities are activated
by early pathological changes in OA. This platform comprised (1) a
peptide linker substrate for metalloproteinase-13 (MMP-13), a pathological
protease upregulated in OA, which was conjugated to (2) a porphyrin
moiety with inherent multimodal imaging, photodynamic therapy, and
drug delivery capabilities, and (3) a quencher that silences the porphyrin’s
endogenous fluorescence and photoreactivity when the beacon is intact.
In diseased OA tissue with upregulated MMP-13 expression, this porphyrin
molecular beacon (PPMMP13B) was expected to undergo sequence-specific
cleavage, yielding porphyrin fragments with restored fluorescence
and photoreactivity that could, respectively, be used as a readout
of MMP-13 activity within the joint for early OA imaging and disease-targeted
photodynamic therapy. This study focused on the synthesis and characterization
of PPMMP13B, followed by a proof-of-concept evaluation
of its OA imaging and drug delivery potential. In solution, PPMMP13B demonstrated 90% photoactivity quenching in its intact
form and robust MMP-13 activation, yielding a 13-fold increase in
fluorescence post-cleavage. In vitro, PPMMP13B was readily
uptaken and activated in an MMP-13 cell expression-dependent manner
in primary OA synoviocytes without exuding significant cytotoxicity.
This translated into effective intra-articular cartilage (to a 50
μm depth) and synovial uptake and activation of PPMMP13B in a destabilization of the medial meniscus OA mouse model, yielding
strong fluorescence contrast (7-fold higher signal than background)
at the diseased joint site. These results provide the foundation for
further exploration of porphyrin molecular beacons for image-guided
OA disease stratification, effective articular delivery of disease-modify
agents, and OA photodynamic therapy.