Evaluation of a polarization-enhanced laparoscopy prototype for improved intra-operative visualization of peritoneal metastases.

Despite careful staging, the accuracy for preoperative detection of small distant metastases remains poor, creating a clinical need for enhanced operative staging to detect occult peritoneal metastases. This study evaluates a polarization-enhanced laparoscopy (PEL) prototype and assesses its potential for label-free contrast enhancement of peritoneal metastases. This is a first-in-human feasibility study, including 10 adult patients who underwent standard staging laparoscopy (SSL) for gastrointestinal malignancy along with PEL. Image frames of all detectable peritoneal lesions underwent analysis. Using Monte Carlo simulations, contrast enhancement based on the color dependence of PEL (mPEL) was assessed. The prototype performed safely, yet with limitations in illumination, fogging of the distal window, and image co-registration. Sixty-five lesions (56 presumed benign and 9 presumed malignant) from 3 patients represented the study sample. While most lesions were visible under human examination of both SSL and PEL videos, more lesions were apparent using SSL. However, this was likely due to reduced illumination under PEL. When controlling for such effects through direct comparisons of integrated (WLL) vs differential (PEL) polarization laparoscopy images, we found that PEL imaging yielded an over twofold Weber contrast enhancement over WLL. Further, enhancements in the discrimination between malignant and benign lesions were achieved by exploiting the PEL color contrast to enhance sensitivity to tissue scattering, influenced primarily by collagen. In conclusion, PEL appears safe and easy to integrate into the operating room. When controlling for the degree of illumination, image analysis suggested a potential for mPEL to provide improved visualization of metastases.

Label-free, multi-parametric assessments of cell metabolism and matrix remodeling within human and early-stage murine osteoarthritic articular cartilage.

Osteoarthritis (OA) is characterized by the progressive deterioration of articular cartilage, involving complicated cell-matrix interactions. Systematic investigations of dynamic cellular and matrix changes during OA progression are lacking. In this study, we use label-free two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging to assess cellular and extracellular matrix features of murine articular cartilage during several time points at early stages of OA development following destabilization of medial meniscus surgery. We detect significant changes in the organization of collagen fibers and crosslink-associated fluorescence of the superficial zone as early as one week following surgery. Such changes become significant within the deeper transitional and radial zones at later time-points, highlighting the importance of high spatial resolution. Cellular metabolic changes exhibit a highly dynamic behavior, and indicate metabolic reprogramming from enhanced oxidative phosphorylation to enhanced glycolysis or fatty acid oxidation over the ten-week observation period. The optical metabolic and matrix changes detected within this mouse model are consistent with differences identified in excised human cartilage specimens from OA and healthy cartilage specimens. Thus, our studies reveal important cell-matrix interactions at the onset of OA that may enable improved understanding of OA development and identification of new potential treatment targets.

Riboflavin depletion promotes longevity and metabolic hormesis in Caenorhabditis elegans.

Riboflavin is an essential cofactor in many enzymatic processes and in the production of flavin adenine dinucleotide (FAD). Here, we report that the partial depletion of riboflavin through knockdown of the C. elegans riboflavin transporter 1 (rft-1) promotes metabolic health by reducing intracellular flavin concentrations. Knockdown of rft-1 significantly increases lifespan in a manner dependent upon AMP-activated protein kinase (AMPK)/aak-2, the mitochondrial unfolded protein response, and FOXO/daf-16. Riboflavin depletion promotes altered energetic and redox states and increases adiposity, independent of lifespan genetic dependencies. Riboflavin-depleted animals also exhibit the activation of caloric restriction reporters without any reduction in caloric intake. Our findings indicate that riboflavin depletion activates an integrated hormetic response that promotes lifespan and healthspan in C. elegans.

Polarization enhanced laparoscope for improved visualization of tissue structural changes associated with peritoneal cancer metastasis.

A polarization enhanced laparoscopy (PEL) imaging system was developed to examine the feasibility of utilizing PEL to augment conventional white light laparoscopy (WLL) in the visualization of peritoneal cancer metastases. The system includes a modified tip to illuminate tissue with linearly polarized light and elements in the detection path enabling recording of corresponding images linearly co- and cross-polarized relative to the incident light. WLL and PEL images from optical tissue phantoms with features of distinct scattering cross-section confirm the enhanced sensitivity of PEL to such characteristics. Additional comparisons based on images acquired from collagen gels with different levels of fiber alignment highlight another source of PEL contrast. Finally, PEL and WLL images of ex vivo human tissue illustrate the potential of PEL to improve visualization of cancerous tissue surrounded by healthy peritoneum. Given the simplicity of the approach and its potential for seamless integration with current clinical practice, our results provide motivation for clinical translation.

Polarization gating technique extracts depth resolved fluorescence redox ratio in oral cancer diagnostics.

Mortality of oral cancer is often due to late diagnosis. Effective non-invasive diagnostic techniques may increase the survival rate based on an earlier diagnosis.. We report on the application of the polarization gating technique for isolating weakly scattered and highly scattered components of fluorescence emission from the superficial and deeper layers of tissue due to intrinsic fluorophores NADH and FAD. The fluorescence polarization spectra were collected from 21 normal and 67 oral squamous cell carcinoma biopsy tissues. The tissues were excited at 350 nm and the fluorescence emission had two peaks corresponding to NADH, and FAD respectively. The spectra were analyzed using the polarization gating technique along with the spectral deconvolution method to derive the optical redox ratio from different layers of tissue. The fractional change in redox ratio between superficial and deeper layers of tissue exhibits excellent statistical significance (p<10-3) which may be due to a shift in the metabolic pathway from oxidative phosphorylation to glycolysis in the cancer cell. Further, variation in collagen intensity in deeper layers of tissue is observed which may be attributed to the breakdown of collagen fibers in the stroma. Linear discriminant analysis showed that oral cancer tissue is discriminated with a better accuracy using polarization gating technique than that of conventional fluorescence spectroscopy.

Synchronous Luminescence Spectroscopy as a Tool in the Discrimination and Characterization of Oral Cancer Tissue.

High incidence of oral cancer is primarily due to ongoing tobacco epidemic. In this work, synchronous luminescence spectroscopy (SLS) has been used to characterize and discriminate oral cancer tissue. Spectral deconvolution method is employed to compute the fluorescence intensity, peak wavelength, and full width half maxima for different endogenous fluorophores. The fluorescence measurements were made on 21 normal and 88 oral squamous cell carcinoma biopsy tissues. Besides, variations in relative concentration of collagen, NADH, and FAD, peak shifts and broadening of peaks are observed for tryptophan, NADH, and FAD, in oral cancer tissues indicating both biochemical and micro environmental changes at cellular level. Linear discriminant analysis showed that oral cancer tissue is discriminated with a sensitivity and specificity of 100% and 95.2% respectively.