Tissue engineered cartilage integration to live and devitalized cartilage: a study by reflectance mode confocal microscopy and standard histology.

This study investigated the in vivo formation of engineering cartilage within living or devitalized cartilage discs using reflectance mode confocal microscopy and conventional light microscopy. Pig articular chondrocytes were suspended in fibrin glue and placed between two cartilage discs. Four experimental groups were prepared: in groups 1 and 2, the cell-hydrogel composite was placed between two live or between two devitalized cartilage discs, respectively; in groups 3 and 4, acellular fibrin glue was placed between two live or between two devitalized cartilage discs, respectively. Samples were implanted in the back of nude mice and analyzed after 2, 5, and 8 weeks. Results showed that engineered cartilage seems to grow more homogenously when the cell-seeded gel was placed between devitalized cartilages than when it was placed between live cartilage matrices. Confocal microscopy provides valuable information on the integration of tissue-engineered cartilage with native tissue and could be useful for nondestructive imaging in vivo.

Virtual biopsy of the joint tissues using near-infrared, reflectance confocal microscopy. A pilot study.

Standard noninvasive imaging techniques applied to joints provide gross morphological features, insufficient for assessing histological detail. On the other hand, biopsying is invasive, time consuming, and may involve unwanted processing artifacts. Near-infrared reflectance confocal microscopy is a technique that allows serial, high-resolution optical sectioning through intact tissues without employing exogenous fluorescent stains. The aim of this work was to evaluate the potential utility of near-infrared reflectance confocal microscopy for providing immediate histological information on meniscus, articular cartilage, epiphyseal plate, bone, muscle, and tendon. Images from near-infrared reflectance confocal microscopy were compared with mirror routine histology sections. Characteristic architectural features were readily visualized in the three dimensions of space. Additionally, the use of experimental contrast agents highlighted the localization of nuclei. Limitations include penetration depth and minor optical artifacts. In conclusion, near-infrared reflectance confocal microscopy is a useful technique for immediate, nondestructive, serial "virtual" sectioning through intact tissues, being thus a potential adjunct to current imaging techniques in orthopedics.

Uremia associated with erythrophagocytosis by small intestine epithelial cells: a case report.

BACKGROUND: Anemia is a frequent finding in patients with uremia due to chronic renal failure. Two factors contribute to the decrease in the red blood cell count and worsen the patient's general status: depression of erythropoiesis and shortening of the red blood cell lifespan. CASE: A novel response mechanism to erythrocyte loss took place in a uremic patient with gastrointestinal hemorrhage. In an autopsy case of an 80-year-old woman dying of uremia, analysis of an intestinal fluid smear revealed small intestine epithelial cells engulfing complete erythrocytes. CONCLUSION: This cytologic finding could account for a potential response mechanism to counteract the massive erythrocyte loss that occurs in hemorrhagic anemia with a uremic background.

Novel virtual biopsy of the kidney with near infrared, reflectance confocal microscopy: a pilot study in vivo and ex vivo.

PURPOSE: Current diagnostic strategies for the kidney combine noninvasive imaging techniques with invasive procedures such as needle biopsy. However, renal needle biopsy is not devoid of risks, such as bleeding or infection. Additionally, histology studies are limited to ex vivo morphology and processing induces tissue artifacts, is time-consuming and limits the performance of further studies. Near infrared, reflectance CM is a novel technique that allows high resolution optical sectioning through intact tissues without using any exogenous fluorescent stains. Contrast between structures is based on the natural differences in refractivity. In this pilot study we assessed the usefulness of CM in the study of the kidney in vivo and ex vivo. MATERIALS AND METHODS: Kidneys of live rats were imaged with CM. Contrast in images is based on native properties of the tissue upon being shone with a near infrared laser. By convention hyperrefractile structures are seen as white and hyporefractile structures are seen as black. CM imaging planes varied along the x, y and z axes of space. Images of live kidney were compared with those from ex vivo CM imaging and standard histology procedures. RESULTS: The tubules, glomeruli, vessels and interstitium were readily identified, revealing intracellular detail. Differences between the kidney in vivo and ex vivo were also observed. Experimental contrast agents further highlighted the nuclei. CONCLUSIONS: CM is a useful noninvasive imaging technique that is an adjunct to current techniques for 3-dimensional study of the kidney in vivo and ex vivo. Future technical developments will provide key applications during surgical intervention, transplantation and rapid tissue assessment.

Evaluation of human knee meniscus biopsies with near-infrared, reflectance confocal microscopy. A pilot study.

Knee cartilage biopsy is used to confirm the pathology in both clinical and experimental conditions and often guides diagnosis and therapeutic strategies. Current histopathological techniques are time consuming, induce tissue artefacts and often prevent further evaluation, once the tissue has been fixed. Hence, there is a potential need for a fast and nondestructive imaging technique for unfixed tissue. Near-infrared, reflectance confocal microscopy (CM) allows real-time, virtual sectioning of unstained, bulk tissue samples. This pilot study evaluates the use of CM in the assessment of meniscus histopathology in a series of 26 freshly-excised human meniscus samples compared to standard light microscopy of stained sections. CM images of the meniscus show cell and matrix detail, depicting morphologic features of collagen and elastic fibres, vessels and nerve endings. In addition, crystal deposits of gout and pseudogout are also demonstrable. Thus, CM is a novel imaging technique that could enable the pathologist to make a rapid microscopic evaluation of cartilage in a fresh and unfixed fashion.

In vivo and ex vivo virtual biopsy of the liver with near-infrared, reflectance confocal microscopy.

The assessment of liver architecture is an essential part of the understanding of its physiology and pathology. Current fluorescence confocal microscopy methods face numerous drawbacks, such as cytotoxicity, quenching effect, potential negative ino- and chrono-tropic effects and leaking of fluorescent agents through the sinusoid fenestrations. The recently developed, near-infrared reflectance confocal microscopy allows high-resolution optical sectioning through intact tissues, without employing fluorescent stains, while contrast between structures is provided by the natural refractivity of the tissue. The aim of this study is to assess the utility of near-infrared reflectance confocal microscopy in the evaluation of the hepatic microscopic architecture in vivo and ex vivo. Rat livers were noninvasively examined in vivo and ex vivo with near-infrared reflectance confocal microscopy. Two experimental contrast agents were subsequently used to enhance particular structures. Parenchymal and vascular structures are readily identified, as well as some intracellular details. Differences between in vivo and ex vivo states were also observed. The use of contrast agents also highlights certain morphologic structures. In conclusion, near-infrared reflectance confocal microscopy stands as a useful adjunct technique to the study of hepatic parenchyma offering details equivalent to, if not surpassing traditional light microscopy.

Evaluation of hepatic histology by near-infrared confocal microscopy: a pilot study.

Liver biopsy is a necessary procedure in establishing the tissue diagnosis of many liver conditions and often guides therapeutic strategies. Current histopathologic techniques are either time-consuming or tissue-destroying; hence the potential need for a fast and nondestructive imaging technique of unfixed human liver. This pilot study evaluates the use of near-infrared reflectance confocal microscopy (CM) in the study of human liver histopathology. Without cutting or staining the tissue, CM provides images of bulk parenchyma showing cellular and subcellular detail and depicting morphologic features of hepatic parenchyma in both diseased and nondiseased states. This article presents a series of 12 human liver biopsy samples, providing an overview on the potential of this technique in assessing common findings from light microscopy.