Comparing PRP and bone marrow aspirate effects on cartilage defects associated with partial meniscectomy: a confocal microscopy study on animal model.

AIM: The aim of our study was to assess the therapeutic effects of platelet-rich plasma (PRP) and bone marrow aspirate concentrate (BMAC) in an animal knee lesion complex associating a large osteochondral defect and meniscal defect resulted from partial meniscectomy, a clinical situation that occurs quite often in orthopedic practice. MATERIALS AND METHODS: Twenty-one male rabbits were included in the study, and all underwent initial surgery on the right knee to create the osteochondral defect on the internal femoral condyle, and remove the anterior horn of the internal meniscus, simulating a clinical situation. Rabbits were separated in three study groups: control, PRP group, in which three PRP injections were administered, and BMAC group, in which one single BMAC injection was administered. At the end of the six months follow-up period, knees were harvested and further analyzed using confocal microscopy and three-dimensional (3D) reconstruction of the articular surface. RESULTS: Therapeutic groups had better results concerning articular surface remodeling and joint degeneration indicators in comparison to trauma group. CONCLUSIONS: Our results suggest that using post-operative regenerative therapies does improve final results concerning surface contact remodeling that was investigated using confocal microscopy and should be considered a valid treatment adjuvant in managing patients with this type of lesion complex, as it improves global joint outcome.

Paving the way to tumor budding assessment using digital pathology: a pilot study in Timisoara City (Romania).

BACKGROUND AND AIM: The outcome for some colorectal cancer patients correlates poorly with classical prognostic factors, like tumor stage. Tumor budding (TB) is a promising and intensely studied new prognostic factor. We aimed to evaluate the reliability of bud counting on Hematoxylin-Eosin (HE)-stained and immunohistochemically (IHC)-stained scanned slides. MATERIALS AND METHODS: We evaluated 21 cases of robotic surgery colorectal cancer specimens that were submitted to the Department of Pathology, Emergency County Hospital, Timisoara, Romania. TB was assessed by one experienced (R3) and two junior pathologists (R1, R2), in 10 circular areas at 20× (0.785 mm²) on scanned HE-stained and IHC-stained [cytokeratin (CK) AE1∕AE3] slides. Interobserver agreement (Cohen's kappa) and intraclass correlation coefficient (ICC) were calculated. RESULTS: In the case of HE-stained slides, the inter-item correlation matrix showed values between 0.632 and 0.84, while the ICC on average measures for consistency showed very good correlation [ICC: 0.887, 95% confidence interval (CI): 0.765-0.95)]. The inter-item correlation matrix for IHC-stained slides comprised values between 0.864 and 0.921, while the ICC for average measures for consistency yielded an excellent value (ICC: 0.95, 95% CI: 0.896-0.978). We identified higher values for budding scores on IHC-stained slides, in comparison to the HE-stained slides: in 19∕21 cases for R1 (average increase of 234.85%), 16∕21 cases for R2 (average increase of 114.14%), and 20∕21 cases for R3 (average increase of 66.92%). CONCLUSIONS: We consider the method of buds counting in 10 microscopic fields on scanned slides to be reliable and valuable. TB counts are higher on IHC-stained slides and associate a better interobserver agreement.

In vivo confocal microscopy quantification of reactive oxygen species: a working model in rat kidney.

Oxidative stress is a culprit responsible for the development of acute and chronic kidney diseases. We aimed to establish a working model for the dynamic in vivo assessment of reactive oxygen species (ROS) production in rat kidney. A randomized controlled study was performed in 36 adult male Wistar rats subjected to unilateral urinary obstruction (UUO) via ureteral ligation and compared to SHAM controls. Dihydroethidium (DHE) was injected in the femoral vein and in vivo confocal microscopy was performed in the 2nd, 6th and 10th day, respectively after surgery. Maximal ROS levels elicited by UUO were recorded on the 6th day. However, the absolute difference of the means of DHE fluorescence intensity between UUO and SHAM was the highest on the 10th day. Our working model can monitor ROS production at different time frames and our initial findings suggest that the surgery-related ROS levels decline after an initial increase in the first days, whereas the ones elicited by chronic ligation continue to raise.