Aim
The shortcomings of today's white light-based colorectal cancer screening are addressed by colonoscopic fluorescence imaging of preneoplastic lesions using targeted fluorescent agents to enhance contrast between the lesion and the surrounding normal colonic epithelium. Approach: We describe the development of Pluronic® nanoparticles of fluorocoxib A (FA), a fluorescent cyclooxygenase-2 (COX-2) inhibitor that enables targeted imaging of inflammation and cancer in numerous animal models, for endoscopic florescence imaging of colonic adenomas.
Conclusions
These results demonstrate that the formulation of FA in Pluronic nanoparticles overcomes a significant hurdle to its clinical development for early detection of colorectal neoplasms by fluorescence endoscopy.
Results
We formulated FA, a fluorescent COX-2 inhibitor, or fluorocoxib negative control (FNC), a nontargeted fluorophore and a negative control for FA, in micellar nanoparticles of FDA approved Pluronic tri-block co-polymer using a bulk solvent evaporation method. This afforded FA-loaded micellar nanoparticles (FA-NPs) or FNC-loaded micellar nanoparticles (FNC-NPs) with the hydrodynamic diameters ( DhDh<math> <mrow><msub><mi>D</mi> <mi>h</mi></msub> </mrow> </math> ) of 45.7±2.5nm45.7±2.5nm<math><mrow><mn>45.7</mn> <mo>±</mo> <mn>2.5</mn> <mtext> </mtext> <mi>nm</mi></mrow> </math> and 44.9±3.8nm44.9±3.8nm<math><mrow><mn>44.9</mn> <mo>±</mo> <mn>3.8</mn> <mtext> </mtext> <mi>nm</mi></mrow> </math> and the zeta potentials ( ζζ<math><mrow><mi>ζ</mi></mrow> </math> ) of −1.47±0.3mV-1.47±0.3mV<math><mrow><mo>-</mo> <mn>1.47</mn> <mo>±</mo> <mn>0.3</mn> <mtext> </mtext> <mi>mV</mi></mrow> </math> and −1.64±0.5mV-1.64±0.5mV<math><mrow><mo>-</mo> <mn>1.64</mn> <mo>±</mo> <mn>0.5</mn> <mtext> </mtext> <mi>mV</mi></mrow> </math> , respectively. We intravenously injected B6;129 mice bearing colonic adenomas induced by azoxymethane and dextran-sodium sulfate with FA-loaded Pluronic nanoparticles (FA-NPs). The diffusion-mediated local FA release and its binding to COX-2 enzyme allowed for clear detection of adenomas with high signal-to-noise ratios. The COX-2 targeted delivery and tumor retention were validated by negligible tumor fluorescence detected upon colonoscopic imaging of adenoma-bearing mice injected with Pluronic nanoparticles of FNC or of animals predosed with the COX-2 inhibitor, celecoxib, followed by intravenous dosing of FA-NPs. Conclusions: These results demonstrate that the formulation of FA in Pluronic nanoparticles overcomes a significant hurdle to its clinical development for early detection of colorectal neoplasms by fluorescence endoscopy.
Significance
Current white light colonoscopy suffers from many limitations that allow 22% to 32% of preneoplastic lesions to remain undetected. This high number of false negatives contributes to the appearance of interval malignancies, defined as neoplasms diagnosed between screening colonoscopies at a rate of 2% to 6%. Aim: The shortcomings of today's white light-based colorectal cancer screening are addressed by colonoscopic fluorescence imaging of preneoplastic lesions using targeted fluorescent agents to enhance contrast between the lesion and the surrounding normal colonic epithelium. Approach: We describe the development of Pluronic® nanoparticles of fluorocoxib A (FA), a fluorescent cyclooxygenase-2 (COX-2) inhibitor that enables targeted imaging of inflammation and cancer in numerous animal models, for endoscopic florescence imaging of colonic adenomas. Results: We formulated FA, a fluorescent COX-2 inhibitor, or fluorocoxib negative control (FNC), a nontargeted fluorophore and a negative control for FA, in micellar nanoparticles of FDA approved Pluronic tri-block co-polymer using a bulk solvent evaporation method. This afforded FA-loaded micellar nanoparticles (FA-NPs) or FNC-loaded micellar nanoparticles (FNC-NPs) with the hydrodynamic diameters ( DhDh<math> <mrow><msub><mi>D</mi> <mi>h</mi></msub> </mrow> </math> ) of 45.7±2.5nm45.7±2.5nm<math><mrow><mn>45.7</mn> <mo>±</mo> <mn>2.5</mn> <mtext> </mtext> <mi>nm</mi></mrow> </math> and 44.9±3.8nm44.9±3.8nm<math><mrow><mn>44.9</mn> <mo>±</mo> <mn>3.8</mn> <mtext> </mtext> <mi>nm</mi></mrow> </math> and the zeta potentials ( ζζ<math><mrow><mi>ζ</mi></mrow> </math> ) of −1.47±0.3mV-1.47±0.3mV<math><mrow><mo>-</mo> <mn>1.47</mn> <mo>±</mo> <mn>0.3</mn> <mtext> </mtext> <mi>mV</mi></mrow> </math> and −1.64±0.5mV-1.64±0.5mV<math><mrow><mo>-</mo> <mn>1.64</mn> <mo>±</mo> <mn>0.5</mn> <mtext> </mtext> <mi>mV</mi></mrow> </math> , respectively. We intravenously injected B6;129 mice bearing colonic adenomas induced by azoxymethane and dextran-sodium sulfate with FA-loaded Pluronic nanoparticles (FA-NPs). The diffusion-mediated local FA release and its binding to COX-2 enzyme allowed for clear detection of adenomas with high signal-to-noise ratios. The COX-2 targeted delivery and tumor retention were validated by negligible tumor fluorescence detected upon colonoscopic imaging of adenoma-bearing mice injected with Pluronic nanoparticles of FNC or of animals predosed with the COX-2 inhibitor, celecoxib, followed by intravenous dosing of FA-NPs. Conclusions: These results demonstrate that the formulation of FA in Pluronic nanoparticles overcomes a significant hurdle to its clinical development for early detection of colorectal neoplasms by fluorescence endoscopy.