Histological Evaluation Research Articles

Role of macroscopic on-site evaluation of endoscopic ultrasound-guided fine-needle aspiration/biopsy: Results of a multicentric prospective study

BACKGROUND The concept of macroscopic on-site evaluation (MOSE) was introduced in 2015 when the endoscopist observed better diagnostic yield when the macroscopically visible core on MOSE was superior to 4 mm. Recent studies suggest that MOSE by the endoscopist may be an excellent alternative to rapid on-site evaluation, and some classifications have been published. Few studies have assessed the adequacy of histologic cores in MOSE during endoscopic ultrasound-guided fine-needle aspiration/biopsy (EUS-FNA/FNB). AIM To evaluate the performance of MOSE during EUS-FNA/FNB. METHODS This multicentric prospective study was conducted in 16 centers in 3 countries (Egypt, Iraq, and Morocco) and included 1108 patients with pancreatic, biliary, or gastrointestinal pathology who were referred for EUS examination. We prospectively analyzed the MOSE in 1008 patients with available histopathological reports according to 2 classifications to determine the adequacy of the histological core samples. Data management and analysis were performed using a Statistical Package for Social Sciences (SPSS) version 27. RESULTS A total of 1074 solid lesions were biopsied in 1008 patients with available cytopathological reports. Mean age was 59 years, and 509 patients (50.5%) were male. The mean lesion size was 38 mm. The most frequently utilized needles were FNB-Franseen (74.5%) and 22 G (93.4%), with a median of 2 passes. According to 2 classifications, 618 non-bloody cores (61.3%) and 964 good samples (95.6%) were adequate for histological evaluation. The overall diagnostic yield of cytopathology was 95.5%. The cytological examination confirmed the diagnosis of malignancy in 861 patients (85.4%), while 45 samples (4.5%) were inconclusive. Post-procedural adverse events occurred in 33 patients (3.3%). Statistical analysis showed a difference between needle types (P = 0.035) with a high sensitivity of FNB (97%). The analysis of the relationship between the MOSE-score and the final diagnosis showed a significant difference between the different scores of the MOSE (P < 0.001). CONCLUSION MOSE is a simple method that allows endoscopists to increase needle passes to improve sample quality. There is significantly higher FNB sensitivity and cytopathology diagnostic yield with good MOSE cores.

Neem seed oil ameliorates diabetic phenotype by suppressing redox imbalance, dyslipidaemia and pro-inflammatory mediators in a rodent model of type 2 diabetes.

The neem plant (Azadirachta indica) has popular ethnomedicinal applications. The anti-diabetic potential and mechanism of neem seed oil (NSO) in a rodent model of type 2 diabetes mellitus was evaluated in the present study. Experimentally-induced diabetic animals were administered NSO (200 and 400 mg/kg) or metformin (150 mg/kg) orally for 30 days, with some animals serving as positive and negative controls. NSO significantly (p < .05) reversed diabetes-induced impaired glucose metabolism, dyslipidaemia, and oxido-inflammatory imbalances typified by changes in the NADH/NAD+ ratio (p < .001) and increases in the mRNA or protein levels of C-reactive protein, 4-hydroxynonenal, and pro-inflammatory cytokines (TNF-α and Il-1β) among others in the hepatic or pancreatic tissues of diabetic animals. The histological evaluation of the pancreatic tissue corroborated the protective effect of NSO. The findings showed that the antidiabetic effect of NSO proceeded through its hypolipidemic effect and modulation of redox and inflammatory signalling events in the tissues of animals.

Multi-Layered Implant Approach for Hemilaryngectomy Reconstruction in a Porcine Model.

Partial laryngectomies result in voice, swallowing, and airway impairment for thousands of patients in the United States each year. Treatment options for dynamic restoration of laryngeal function are limited. Thus, there is a need for new reconstructive approaches. Here, we evaluated early (4 week) outcomes of multi-layered mucosal-myochondral (MMC) implants when used to restore laryngeal form and function after hemilaryngectomy in a porcine model. Six Yucatan minipigs underwent transmural hemilaryngectomies followed by reconstruction with customized MMC implants aiming to provide site-appropriate localization of regenerated laryngeal tissues, while supporting laryngeal function. All implants were fabricated from polymeric collagen, with a subset of muscle and cartilage implants containing motor endplate-expressing muscle progenitor cells or cartilage-like cells differentiated from adipose stem cells, respectively. Vocalization and laryngeal electromyography (L-EMG) measurements with nerve conduction studies were performed post-operatively and compared with baseline along with gross and histological analyses of the healing response. All animals (n = 6) survived and maintained airway patency, safe swallowing, and phonation, without the use of tracheostomy and/or gastrostomy tubes. Histological evaluation indicated no adverse tissue reaction or implant degradation, showing progressive regenerative remodeling with mucosa reformation and ingrowth of new muscle and cartilage. Preliminary L-EMG suggested weak but detectable motor unit action potentials. Although vocalization duration, frequency, and intensity decreased post-operatively, all animals retained vocal capacity and parameter recovery was evident over the study duration. Engineered collagen polymeric implants in the presence or absence of autologous cell populations may serve as a feasible reconstructive option to restore dynamic function after hemilaryngectomy. Long-term follow-up is needed to further assess functional outcomes. NA Laryngoscope, 2024.

Enhanced skin cancer diagnosis: a deep feature extraction-based framework for the multi-classification of skin cancer utilizing dermoscopy images

Skin cancer is one of the most common, deadly, and widespread cancers worldwide. Early detection of skin cancer can lead to reduced death rates. A dermatologist or primary care physician can use a dermatoscope to inspect a patient to diagnose skin disorders visually. Early detection of skin cancer is essential, and in order to confirm the diagnosis and determine the most appropriate course of therapy, patients should undergo a biopsy and a histological evaluation. Significant advancements have been made recently as the accuracy of skin cancer categorization by automated deep learning systems matches that of dermatologists. Though progress has been made, there is still a lack of a widely accepted, clinically reliable method for diagnosing skin cancer. This article presented four variants of the Convolutional Neural Network (CNN) model (i.e., original CNN, no batch normalization CNN, few filters CNN, and strided CNN) for the classification and prediction of skin cancer in lesion images with the aim of helping physicians in their diagnosis. Further, it presents the hybrid models CNN-Support Vector Machine (CNNSVM), CNN-Random Forest (CNNRF), and CNN-Logistic Regression (CNNLR), using a grid search for the best parameters. Exploratory Data Analysis (EDA) and random oversampling are performed to normalize and balance the data. The CNN models (original CNN, strided, and CNNSVM) obtained an accuracy rate of 98%. In contrast, CNNRF and CNNLR obtained an accuracy rate of 99% for skin cancer prediction on a HAM10000 dataset of 10,015 dermoscopic images. The encouraging outcomes demonstrate the effectiveness of the proposed method and show that improving the performance of skin cancer diagnosis requires including the patient's metadata with the lesion image.

Abstract 4118949: Factors Influencing Left Ventricular Function Recovery Following Left Ventricular Assist Device Implantation in Pediatric Dilated Cardiomyopathy: Insights from Single-Cell RNA Sequencing Analysis

Background and Objective: Heart transplantation is a recognized treatment for pediatric severe heart failure, but long-term issues necessitate alternative treatments. Some dilated cardiomyopathy (DCM) cases show left ventricular function recovery after LVAD implantation. This occurrence could lead to new treatments, but mechanisms are unclear. This study explores preoperative molecular factors interacting with LVAD's effects using single-cell transcriptomics on clinical samples. Methods: Twenty-five pediatric DCM patients underwent Berlin Heart EXCOR (BHE) implantation at our institution since 2013. Those with left ventricular ejection fraction (LVEF) improving by &gt;20% were in the Recovery group (R), others in Non-Recovery (NR). Single-cell RNA-sequence was performed on myocardial tissue from six patients (three per group). Differential gene expression and pathway analysis were conducted for each cell type. Findings were validated by comparing them with histological evaluation or clinical examination in all 25 cases. Results: R: 10 patients; NR: 15. R's weight was 6.3 kg, NR's 9.9 kg (p=0.049); LVEF: R 23%, NR 16.5% (p=0.61). In pseudobulk analysis of cardiomyocytes, upregulated genes in R included NPPB, MYL7, PCDH9. KEGG analysis indicated activation of Tight Junction and ECM receptor pathways in R. In fibroblasts, major extracellular matrix proteins were higher in R, GSL gene expression in NR. Plasma Brain Natriuretic Peptide (BNP) at BHE implantation was significantly higher in R; 2913 [1836-3990 (95% CI)] vs 1278 [667-1888] pg/ml (Image1,2). No differences in tissue fibrosis rates were found between the groups. Conclusion: Alterations in gene expression patterns in some cell clusters, including cardiomyocytes, may contribute to left ventricular function recovery post-BHE implantation in pediatric DCM patients. Plasma BNP levels correlated with gene expression alterations, suggesting both a potentially protective effect on cardiomyocytes and a useful marker of LV functional recovery. Further validation of these candidate genes may elucidate intracellular molecular mechanisms.

Kirschner wire creates more microdamage than standard or acrylic drill bits in the rabbit (Oryctolagus cuniculi) femur.

Histologically evaluate damage to rabbit femur after the creation of bicortical 1.5-mm-diameter holes using a standard surgical drill bit, an acrylic drill bit, and a Kirschner wire (K-wire). 10 femora (5 pairs) from skeletally mature female intact New Zealand white rabbits were used. The bone diaphyses were divided into 4 locations, systematically undergoing each test (surgical drill bit, acrylic drill bit, K-wire, and intact control). Four pairs were drilled using a mechanical testing machine, and 1 pair was drilled by hand. Cross-sections of the bone were stained en bloc with basic fuchsin for undecalcified histological evaluation. Damaged bone was reported as a percentage of a standardized area and categorized by location (cis- or transcortex), drill contact (entrance or exit of the cortex), and total damage (both cortices). The drilling method (hand vs mechanical testing machine) had no effect on histologic damage, so results were analyzed by combining all data. The K-wire demonstrated the greatest area of cracks/damage compared to both standard surgical and acrylic drill bits, whereas no difference in damage was noted between the 2 drill bits for all variables. The K-wire and drill bits caused microdamage; K-wire drilling created more microdamage than drill bits. The rabbit bone cortex is thin and brittle relative to dogs and cats, leading to failure during and after fracture fixation. The clinical failure of rabbit bone is at least partially caused by drill bits or K-wires causing microcracks.

Amelioration of propionic acid-induced autism-like behaviors in rats by fenofibrate: A focus on reduction of brain galectin-3 levels.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by impaired social interactions and repetitive behaviors. This study examines the effects of fenofibrate on a propionic acid (PPA)-induced rat model of ASD, focusing on behavioral changes, inflammatory markers, and histological findings. Thirty male Wistar rats were divided into three groups: a control group, a group receiving PPA and saline, and a group treated with PPA and fenofibrate for 15 days. Behavioral assessments, including the three-chamber sociability test, open-field test, and passive avoidance learning, were conducted. Biochemical analyses measured TNF-α, NGF, IL-17, IL-2, and galectin-3 levels in brain tissues. Histological evaluations focused on Purkinje neuron counts in the cerebellum and neuronal changes in the CA1 and CA3 regions of the hippocampus, along with glial fibrillary acidic protein (GFAP) levels. Fenofibrate treatment significantly improved behavioral outcomes, reducing autism-like behaviors compared to the PPA/saline group. Biochemically, the PPA/saline group showed elevated levels of malondialdehyde, TNF-α, IL-2, IL-17, and galectin-3, which were reduced following fenofibrate treatment. Histologically, the PPA/saline group exhibited fewer, dysmorphic Purkinje neurons and increased glial activity in the CA1 region, both of which were ameliorated by fenofibrate treatment. Fenofibrate shows promise in mitigating autism-like behaviors in a rat model of ASD, likely due to its antioxidative and neuroprotective properties, which contribute to preserving neuronal integrity and reducing inflammation.

Neutrophils enhance the clearance of systemic amyloid deposits in a murine amyloidoma model

IntroductionAmyloid-specific antibodies have been shown to opsonize and enhance amyloid clearance in systemic amyloidosis mouse models. However, the immunological mechanisms by which amyloid is removed have not been clearly defined. Previous reports from preclinical in vivo studies suggest polymorphonuclear cells (i.e., neutrophils) can affect amyloid removal. Therefore, we sought to analyze how neutrophils may contribute to the clearance of human AL amyloid extracts, using a murine amyloidoma model.MethodsImmunocompromised nude mice injected subcutaneously with patient-derived AL amyloid extract (generating a localized “amyloidoma”) were used to circumvent confounding factors contributed by the adaptive immune system and served as the model system. Two representative AL amyloid extracts were used, ALλ(CLA), which is refractory to clearance, and ALκ(TAL), which is readily cleared in mice. Neutrophil recruitment to the amyloid masses, cellular activation, and propensity to engulf amyloid were assessed.ResultsImmunophenotyping of amyloidomas from animals implanted with 2 mg of either ALλ or ALκ revealed that more neutrophils were recruited to ALκ amyloid masses as compared to the ALλ material, which was generally devoid of neutrophils. Ex vivo analyses indicated neutrophils do not efficiently phagocytose amyloid directly. However, histological evaluation of the ALκ amyloidoma revealed the abundant presence of neutrophil extracellular traps, which were absent in the ALλ amyloidomas. Using neutrophil depletion experiments in mice, we determined that mice devoid of neutrophils cleared the human amyloid lesions less efficiently. Moreover, mice devoid of neutrophils also had significantly reduced intra-amyloid expression of inflammatory cytokines.DiscussionNeutrophils may not directly mediate amyloid clearance through phagocytosis; however, these cells can be stimulated by the amyloid and may function to facilitate phagocytosis and amyloid clearance by professional phagocytes (e.g., macrophages).

SIRT1-Dependent Neuroprotection by Resveratrol in TOCP-Induced Spinal Cord Injury: Modulation of ER Stress and Autophagic Flux

This study explores the neuroprotective effects of resveratrol (Resv) against tri-o-cresyl phosphate (TOCP)-induced neurotoxicity in the spinal cord of adult hens. It is well documented that TOCP exposure causes significant neurodegeneration via mechanisms that involve endoplasmic reticulum (ER) stress and impaired autophagy. In this experiment, adult hens were assigned to one of four groups: Control, Resv, TOCP, and TOCP + Resv. The spinal cord tissues were examined through transmission electron microscopy, hematoxylin and eosin (HE) staining, Nissl staining, and Western blotting to evaluate key proteins associated with ER stress and autophagy. Additionally, RT-qPCR and immunofluorescence were employed to measure sirtuin1 (SIRT1) expression. The findings revealed that TOCP induced severe ultrastructural damage, including disrupted myelin sheaths, dilated ER, and extensive neurodegeneration, as confirmed by histological evaluations. The expression levels of GRP78, p-PERK, p-eIF2α, ATF4, CHOP, Beclin-1, P62, and LC3-II were also significantly elevated by TOCP. However, Resv treatment markedly attenuated these pathological changes by reducing ER stress, restoring autophagic flux, and upregulating SIRT1 expression, preserving spinal cord integrity. These results indicate that Resv can effectively counteract TOCP-induced neurotoxicity by modulating ER stress and autophagy, underscoring its potential as a therapeutic agent for neuroprotection.

INNV-20. FLUORESCEIN GUIDED SURGERY FOR ATYPICAL BRAIN LESIONS: IS IT USEFULL? CASE SERIES USING A LOW-COST DEVICE

Abstract INTRODUCTION Sodium Fluorescein (SF) enhances in areas in the brain with a disrupted Blood brain barrier (BBB). In neurosurgery, SF is extensive used for malignant brain tumors. Solitary brain abscesses (BA) are characterized by the pathognomonic finding of BBB disruption. Consequently, FL may have the potential to improve the intra-operative visualization of BA. Other infections also disrupt BBB and SF may guide these surgeries. CASE SERIES A 32-year old male patient, previously diagnosed with HIV presented with headache and during investigation MRI disclosure an extensive leptomeningeal commitment. Leptomeningeal biopsy guided by fluorescein and using Varioguide system was performed and the diagnosis was primary central nervous system lymphoma (Diffuse large B-cell). During surgery, it was observed increased fluorescence in arachnoid. A 72-year old woman presented with headache, impairment of mental status and epilepsy and a KPS of 60. Her initial hypothesis was metastasis but patient did not have a primary situs diagnosis. An open biopsy was performed through temporal posterior craniotomy. During surgery no visible well delimited tumor was found, even with ultrasound use. Fluorescein guided biopsy was performed. Histological evaluation showed an Epstein-Barr virus infection causing encephalitis. This is an extremely rare presentation of this type of infection and SF was extremely helpful to guide the procedure. CONCLUSION This paper reported atypical uses of SF for brain mass lesions surgeries. One of our cases is a unique presentation of a virus encephalitis which mimicked metastasis and other of increased fluorescence in Lymphoma. These atypical use of SF are relevant for further neurosurgical plannings Keywords: Fluorescein, Lymphoma, Biopsy

NCOG-19. FUNCTIONAL OUTCOME AFTER RE-RESECTION OF TUMOR PROGRESSION, MIXED LESION OR TREATMENT-ASSOCIATED EFFECTS IN IRRADIATED DIFFUSE GLIOMAS (GRADE 2-4): IDENTIFYING CLINICAL AND RADIOLOGICAL DETERMINANTS

Abstract In cases of radiological progression in patients with irradiated diffuse gliomas, recurrence-resection is frequently performed. In our previous histological evaluation 19% of these re-resection cases were found to be due to treatment-associated effects (TAE). Limited literature substantiating which patients will benefit from surgery and which patients will deteriorate postoperatively is available. We aim to (a) describe the functional outcomes after re-resection of an irradiated diffuse glioma in adult patients; (b) compare early functional outcomes between patients with TAE versus progressive disease; (c) identify predictors of early functional outcome. In this single-center, retrospective, cohort study, we included consecutive adult patients with diffuse glioma who underwent first-line radiotherapy and, upon radiological progression, a re-resection. Re-resection tissue was classified histologically (based on mitosis presence/absence) and by the multidisciplinary tumor board as tumor progression, TAE or mixed lesion. The primary endpoint was the functional outcome 30 days postoperatively, expressed as the Karnofsky Performance Scale (KPS). Univariable and multivariable logistic regression were used to identify clinicoradiological predictors of KPS deterioration of any severity. Of 159 included patients, 41 (25.8%) experienced mild KPS deterioration (10 points) and 25 (15.9%) a clinically relevant KPS deterioration (20 or more). KPS improvement (10 points or more) occurred in 31 patients (19.4%). Histological diagnosis groups did not differ significantly in KPS outcome. Pre-operative steroid usage (any dose) was an independent predictor of KPS deterioration (OR 2.46, 95%CI 1.12-5.39, p=0.025). Analysis of pre-operative radiological determinants is in progress. In conclusion, in this large single-center cohort of diffuse gliomas undergoing re-resection with systematic clinical evaluation, clinically relevant deterioration after re-resection occurred in 15.9% of all patients. Risk of deterioration is comparable in all histological diagnosis groups. Pre-operative steroid usage was identified as predictor for deterioration (of any severity). These results may provide clinical guidance in identifying which patients will benefit from re-resection.

Morphological and Histological Changes Following Triamcinolone Injection Alone or Mixed Injection of Triamcinolone and Botulinum Toxin in Upper Eyelid.

The purpose of this experiment was to evaluate the functional and histologic changes in the upper eyelid muscles after injection of triamcinolone acetonide (TA) alone or TA combined with botulinum toxin A (Botox or BTXA) in the cynomolgus monkey model. Twenty eyes of 10 cynomolgus monkeys were divided into 4 groups: 3 experimental groups (1, 2, and 3) and the control group (group 4) based on the injection type. In group 1, 0.5mL of TA (Kenalog 40mg/mL) was administered subconjunctivally (between the conjunctiva and the Müller muscle) with a 26 G needle to the inverted upper eyelid of one eye. In group 2, the same procedure was done with 0.5mL TA injected into the other upper eyelid. After that, 5UI/0.1mL Botox was injected transcutaneously into the suborbicularis oculi space in the middle third of the upper eyelid 3mm above the center of the superior tarsal border. In group 3, a subconjunctival of 0.5mL TA was administered to the inverted upper eyelid of one eye 3 times: the injection day, 2 weeks, and 6 weeks after the first injection, whereas a normal saline injection of the same volume was administered one time to the other eye at the injection day in group 4 (control group). Follow-up was done to evaluate the clinical changes in eyelid position at 2, 6, and 12 weeks after injection. Hematoxylin-eosin and Masson trichrome were used to assess the levator or Muller muscle histology and measure the fiber diameter. During the clinical follow-up, there were no major complications observed in any monkeys. The macroscopic appearance of the upper lid on the biopsy day did not differ among groups 1, 2, and 4. There were no cases in these groups that had subconjunctival TA deposit 3 months after injection. Conversely, in group 3, there were 3/5 eyes showing the subconjunctival TA deposit at the last examination.No specific changes in the marginal reflex distance 1, marginal reflex distance 2 (MRD2), and lid crease were noted in either TA alone injection groups 1, 3, and 4. In contrast, there was a significant decrease in marginal reflex distance 1 at 2 weeks (P = 0.003) and 6 weeks (P = 0.005) after TA injection in group 2 in comparison to the baseline.In terms of MRD2, while in group 2, there were significant differences between the pre-MRD2 and the post-MRD2 till the 2 weeks after injection (P = 0.006), then it became insignificant from the sixth week afterward. In contrast, at 2 weeks after injection, MRD2 was reduced in both TA-injected groups 1 and 3, but the observed difference was not significant in both groups. At 6 and 12 weeks, MRD2 fluctuation was not remarkable in these 2 groups and there were no significant differences in comparison to the baseline (P > 0.05).Histological evaluation showed that Müller muscle does not attach directly to the superior border of the tarsus, but it changes to the tendon before attaching to the tarsal plate. In addition, there were no statistical differences in levator muscle fiber diameter and Müller muscle fiber diameter between the 4 groups, with P = 0. 621 and P = 0.695, respectively. Triamcinolone acetonide combined with BTXA showed better results in decreasing upper eyelid height than TA alone in normal monkey eyelids due to its predictable effect. In addition, there were no differences between the side effects and the histology results between the 4 groups. Therefore, TA combined with BTXA may become a promising treatment for selective thyroid eyelid retraction and could offer an alternative to surgery and its complications.

Characterization of Gingival Epulis Lesions: A Histological and Morphological Approach

Objective: To conduct a clinical and histopathological study of Epulis at Bhittai medical and Dental College, Mirpurkhas. Methodology: A descriptive cross-sectional study was carried out at the Bhittai medical and Dental College Mirpurkhas. Patients clinically diagnosed with gingival outgrowth, across all age groups and both genders, were included. A provisional diagnosis of epulis was made based on clinical examination. After taking a careful history, relevant clinical features were recorded, and a biopsy procedure was performed using infiltration near the lesion or regional nerve block techniques. The data was documented using a pre-designed proforma and analyzed using SPSS version 22.0. Results: Out of 50 subjects, 30 (60%) were female and 20 (40%) were male. Most cases involved the maxillary gingiva (anterior or posterior) (62%). The majority of lesions measured between 2cm and 3cm (76%). Lesions typically exhibited a soft consistency (70%), with fewer cases being firm (8%) or hard (22%). Histologically, peripheral giant cell granuloma (PG) was most prevalent (50%), followed by fibrous epulis (42%), peripheral ossifying fibroma (4%), and peripheral giant cell granuloma with features of central giant cell granuloma (PGCG) (4%). No cases of congenital epulis or pregnancy tumor were reported. Histological evaluation revealed no significant gender differences (p-value 0.86). Conclusion: Clinically, most epulis lesions presented as painless, soft, coral pink masses, typically measuring approximately 2-3 cm in diameter and exhibiting a sessile form and observed more commonly in females, with a predominant occurrence in the maxillary region. Histopathological examination revealed that the most frequent types were fibrous epulis and peripheral giant cell granuloma.

Adipose tissue-derived injectable products combined with platelet-rich plasma for the treatment of osteoarthritis: the promising preclinical results are not confirmed by the clinical evidence.

The association of adipose tissue-derived injectable products with platelet-rich plasma (PRP) has been promoted for osteoarthritis (OA) treatment. The aim of this study was to investigate the preclinical and clinical evidence supporting the potential of this combined approach to treat OA. A systematic review was performed in January 2024 on five databases (PubMed, Embase, Scopus, Cochrane, and Web-of-Science) to identify preclinical in vivo and clinical studies. Safety, OA biomarker changes, and outcomes in terms of clinical and imaging results were analyzed. The quality of studies was assessed with the SYRCLE's tool for preclinical studies and the Downs and Black checklist for clinical studies. Ten preclinical studies (223 animals) and 14 clinical studies (594 patients) were included. Preclinical results documented improvements at the cartilage histological and immunohistochemical evaluation and at the biomarkers level. Clinical studies confirmed the procedure's safety, and the case series suggested satisfactory results in different joints in terms of symptoms and function improvement, with positive findings at the biomarker level. However, the randomized controlled trials did not document any clinical benefit, nor any changes in the imaging analysis. A large heterogeneity and overall poor quality were documented in both preclinical and clinical studies. There is an increasing interest in the use of adipose tissue-derived injectable products associated with PRP for the treatment of OA joints, with preclinical studies showing promising results with this combined approach. However, clinical studies did not confirm the benefits offered by PRP augmentation to adipose tissue-derived injectable products in patients affected by OA.

Immunohistochemical Investigation of Cyclooxygenase-2 Expression in Rabbit Uterine Adenocarcinoma and the Potential Use of COX-2 Inhibitors in Cancer Therapy

Cyclooxygenase-2 (COX-2) is overexpressed in many human and animal cancers. Selective COX-2 inhibitors have shown antitumoral effects in tumors with a high expression of COX-2. This study evaluates (1) the expression of COX-2 in rabbit uterine adenocarcinomas, (2) the correlation between immunophenotypic expression and histopathological changes, and (3) the post-surgery response to therapy with COX-2 inhibitors. Forty rabbit uteri were divided into three groups: neoplastic, hyperplastic, and normal endometrium. A histological and immunohistochemical score was applied to investigate the tumor’s grade and the COX-2 expression. By histological evaluation, 30 cases of endometrial adenocarcinoma, 5 cases of endometrial hyperplasia and 5 normal endometria were found. Of the six cases of endometrial adenocarcinoma with follow-up available, four received a post-surgical treatment with meloxicam and two were treated by surgery alone. The survival time of the animals treated with meloxicam was longer than that observed in the untreated animals. A statistically significant difference in COX-2 IHS was observed between non-neoplastic endometrium and adenocarcinoma. The progressive increase in COX-2 expression from normal epithelium to carcinoma suggests that upregulation of COX-2 expression may play a role in tumor initiation and progression. Our findings suggest the possible use of COX-2 inhibitors in treating uterine adenocarcinoma in rabbits. Further study will be needed to confirm this hypothesis.

Magnesium Membrane Shield Technique for Alveolar Ridge Preservation: Step-by-Step Representative Case Report of Buccal Bone Wall Dehiscence with Clinical and Histological Evaluations

Background/Objectives: Despite the increased use of new resorbable magnesium membranes, there are no reported cases or studies on the use of resorbable magnesium membranes in combination with bone grafts for alveolar ridge preservation (ARP) in cases with severe buccal bone wall dehiscence. This case report aimed to evaluate the effectiveness of the magnesium membrane shield technique in conjunction with bone grafting for ARP, assessing both clinical outcomes and histological bone regeneration. Methods: A 44-year-old female patient presented with a vertical fracture on tooth 24 (FDI Notation System) accompanied with complete destruction of the buccal bone wall. The treatment plan included tooth extraction, ARP using a combination of anorganic bovine bone and autologous bone grafting, and the application of a magnesium membrane as a shield to the pre-existing buccal wall. Six months post-procedure, a bone biopsy was taken from the implant site using a trephine bur. Results: Clinical and radiological evaluations six months after the procedure demonstrated sufficient bone volume for implant placement. Additionally, in the next three months, soft tissue conditioning with a provisional crown resulted in an aesthetically and functionally satisfactory outcome. Histological analysis of the bone biopsy revealed well-formed new bone in direct contact with residual biomaterial, with no signs of inflammation. Osteocytes were clearly visible within the newly formed bone matrix, indicating successful bone maturation. Active osteoblasts were observed along the bone-biomaterial interface, suggesting ongoing bone remodeling and integration. Additionally, histomorphometric evaluation revealed 47% newly formed bone, 32% soft tissue, and 19% residual biomaterial. Conclusions: This case demonstrates the potential of the magnesium shield technique as an ARP technique in cases with severe buccal wall dehiscence. The technique yielded satisfactory clinical outcomes and promoted successful bone regeneration, as confirmed by histological analysis.

A novel approach to engineering three-dimensional bladder tumor models for drug testing.

Bladder cancer (BCa) poses a significant health challenge, particularly affecting men with higher incidence and mortality rates. Addressing the need for improved predictive models in BCa treatment, this study introduces an innovative 3D in vitro patient-derived bladder cancer tumor model, utilizing decellularized pig bladders as scaffolds. Traditional 2D cell cultures, insufficient in replicating tumor microenvironments, have driven the development of sophisticated 3D models. The study successfully achieved pig bladder decellularization through multiple cycles of immersion in salt solutions, resulting in notable macroscopic and histological changes. This process confirmed the removal of cellular components while preserving the native extracellular matrix (ECM). Quantitative analysis demonstrated the efficacy of decellularization, with a remarkable reduction in DNA concentration, signifying the removal of over 95% of cellular material. In the development of the in vitro bladder cancer model, muscle invasive bladder cancer patients' cells were cultured within decellularized pig bladders, yielding a three-dimensional cancer model. Optimal results were attained using an air-liquid interface technique, with cells injected directly into the scaffold at three distinct time points. Histological evaluations showcased characteristics resembling in vivo tumors derived from bladder cancer patients' cells. To demonstrate the 3D cancer model's effectiveness as a drug screening platform, the study treated it with Cisplatin (Cis), Gemcitabine (Gem), and a combination of both drugs. Comprehensive cell viability assays and histological analyses illustrated changes in cell survival and proliferation. The model exhibited promising correlations with clinical outcomes, boasting an 83.3% reliability rate in predicting treatment responses. Comparison with traditional 2D cultures and spheroids underscored the 3D model's superiority in reliability, with an 83.3% predictive capacity compared to 50% for spheroids and 33.3% for 2D culture. Acknowledging limitations, such as the absence of immune and stromal components, the study suggests avenues for future improvements. In conclusion, this innovative 3D bladder cancer model, combining decellularization and patient-derived cells, marks a significant advancement in preclinical drug testing. Its potential for predicting treatment outcomes and capturing patient-specific responses opens new avenues for personalized medicine in bladder cancer therapeutics. Future refinements and validations with larger patient cohorts hold promise for revolutionizing BCa research and treatment strategies.

Self-assembling peptide hydrogel scaffold accelerates healing of patellar tendon injury: A histological and biomechanical study.

Although KI24RGDS peptide hydrogel that acts as a cell adhesion has been reported to repair tissue in meniscus injury, its effect on tendon injuries remains unknown. The purpose of this study was to clarify the effect of KI24RGDS for tendon repair based on histological and biomechanical evaluation. After introducing defects (length: 10mm; width: 3mm) at the centers of rabbits' patellar tendons, and the KI24RGDS group was implanted with KI24RGDS and observed for 8 weeks. KI24RGDS implantation resulted in limited tendon elongation and better histological scores with uniformed collagen fiber orientation and early vascularization. The failure load of the patellar tendon was higher in the KI24RGDS group than that in the defect group (p < 0.05) and no significant difference with the control group (intact patellar tendon) at 8weeks postoperatively. In conclusion, KI24RGDS administration might have therapeutic potential for tendon injuries by accelerating collagen remodeling.

Digital spatial profiling for pathologists.

The advent of "omics" technologies for high-depth tumor profiling has provided new information regarding cancer heterogeneity. However, a bulk omics profile can only partially reproduce tumor complexity, and it does not meet the preferences of pathologists used to perform an in situ assessment of marker expression, for instance, with immunohistochemistry. The NanoString GeoMx® Digital Spatial Profiler (DSP) is a platform for morphology-guided multiplex profiling of tissue slides, which allows the digital quantification of target analytes in different neoplastic settings. To illustrate the feasibility and opportunities offered by DSP from a pathologist's perspective, we applied DSP in three different representative neoplastic settings: breast carcinoma, thyroid anaplastic carcinoma, and biphasic mesothelioma. Because of the perfect overlap between the hematoxylin-eosin-stained slide and the GeoMx areas of interest, in breast carcinoma, two different antibodies allowed the distinction of the tumor cells from the surrounding tumor microenvironment. In biphasic mesothelioma, we could distinguish the epithelioid from the sarcomatoid neoplastic component, and in the thyroid, we easily separated the anaplastic areas from the well-differentiated carcinoma. DSP is a promising tool that combines traditional histological evaluation, allowing spatial assessment of a tumor and its surroundings, and innovative in situ digital profiling. Pathologists should not miss the opportunity to combine morphological and genomic analyses and be at the forefront of investigating the progression of dysplasia/neoplasia, low-grade or high-grade, epithelial/mesenchymal, and, more in general, overcoming the concept of in situ vs. bulk genomic methods.

Effects of Losartan and Fisetin on Microfracture-Mediated Cartilage Repair of Ankle Cartilage in a Rabbit Model.

Microfracture is one surgical treatment strategy for osteochondral lesions of the talus (OLTs) but results in fibrocartilage repair tissue, which has inferior mechanical properties to native hyaline cartilage. Biological regulation of microfracture has been suggested to improve the quality of cartilage repair in patients. To determine if administration of losartan, fisetin, or losartan and fisetin combined can enhance microfracture-mediated cartilage repair of OLTs in a rabbit model. Controlled laboratory study. Four-month-old female rabbits were divided into the following groups (8 rabbits per group): microfracture only (microfracture), microfracture plus losartan (losartan), microfracture plus fisetin (fisetin), and microfracture plus losartan and fisetin (losartan+fisetin). A 2.7-mm osteochondral defect and 4 microfracture holes were created in the talar dome cartilage. The rabbits were administered losartan (10 mg/kg/day), fisetin (20 mg/kg/day), or losartan and fisetin orally until euthanized 12 weeks after surgery. Gross evaluation, micro-computed tomography, histology, and immunohistochemistry evaluations of the osteochondral defects were performed as well as quantitative polymerase chain reaction of capsule tissue and enzyme-linked immunosorbent assay of serum. The losartan and fisetin groups had increased International Cartilage Regeneration & Joint Preservation Society macroscopic scores with improved cartilage repair and enhanced subchondral bone healing compared with the microfracture group. However, the losartan+fisetin group did not show a synergistic effect. O'Driscoll histology scores were higher in the losartan and fisetin groups compared with the microfracture group, while the losartan+fisetin group had a lower score than the losartan, fisetin, and microfracture groups. Collagen type 2 staining revealed organized chondrocytes in the losartan and fisetin groups, but the losartan+fisetin group did not show improvement when compared with other groups. Fisetin treatment decreased catalase and transforming growth factor-β1-activated kinase 1 expression in capsular tissue. Concomitant microfracture and biological regulation, using oral administration of either losartan or fisetin, may improve cartilage healing of OLTs; however, losartan and fisetin combined in the current drug administration regimen does not appear to provide synergistic effects. Oral intake of losartan or fisetin may result in beneficial effects on microfracture-mediated cartilage repair of OLTs.