Comparative assessment of bone regeneration by histometry and a histological scoring system / Evaluarea comparativă a regenerării osoase utilizând histometria și un scor de vindecare histologică

Obiective: Scopul studiului de față a constat în evaluarea valorii scorului de vindecare histologică, comparativ cu histometria în monitorizarea vindecării osose la nivelul calotei. Material și metodă: Am realizat un studiu cazcontrol cu un lot control și unul de studiu. La un număr de 60 de șoareci CD1 incluși în lotul de studiu am indus chirurgical un defect osos la nivelul calotei și am realizat reconstrucția defectului utilizând grefe obținute prin inginerie tisulară. Ingineria tisulară a grefonului osos s-a realizat utilizând celule stem embrionare poziționate pe suport matriceal -corn caduc de cerb, iar ca inductor al diferențierii am utilizat mediu osteogenetic bazal și complex. La cei 30 de șoareci CD1 incluși în lotul control am indus chirurgical același defect osos la nivelul calotei, dar nu am realizat reconstrucția osoasă a acestuia. Procesul de regenerare osoasă a fost evaluat la 2 și respectiv la 4 luni utilizând scorul de vindecare și histometria. Rezultate: Scorul de vindecare histologică s-a corelat statistic semnificativ cu dimeniunea defectului obtinută la histometrie (p<0.001). Evaluarea parametrilor în baza cărora s-a stabilit scorul de vindecare histologică indică regenerarea cea mai avansată la subiecții din lotul de studiu sacrificați la 4 luni, la care s-a utilizat pentru ingineria grefonului osos celule stem embrionare, suport matriceal corn caduc de cerb și mediu osteogenetic complex ca inductor. Concluzii: scorul de vindecare histologică este o metoda valoroasă de cuantificare a procesului de regenerare osoasă. Relevanță clinică: Acest studiu demonstrează că scorul de vindecare histologică prezentat este un instrument util pentru clinician în procesul de evaluare a regenerării osoase

The value of a new score for sonoelastographic differentiation between benign and malignant cervical lymph nodes

Abstract Objective: The aim of this study is to explore the diagnostic value of sonoelastography for the differentiation between benign and malignant superficial lymph nodes of the neck. In this respect the utility of an original scoring system was explored. Material and method: Over a period of 30 months the patients examined routinely for the assessment of superficial lymph nodes of the neck were recorded in a data base containing grey-scale, Doppler and sonoelastographic information and images. The sonoelastographic images of 30 benign and 39 malignant lymph nodes were assessed. The images were scored according to a new, eight pattern scoring system proposed by our group. Interobserver agreement and area under the ROC curve (AUROC) for the differentiation between benign vs. malignant and benign vs. metastatic nodes were analyzed. Results: The analysis of the interobserver agreement for the investigated score provided a weighted Kappa = 0.687, 95%CI [0.572 to 0.802] and standard error = 0.059. In the differentiation benign -malignant, the AUROC was 0.846, with sensitivity of 66.67% and specificity of 96.67% for score &gt;3. In the differentiation between benign and metastasis, the same criterion provided an AUROC of 0.855, with sensitivity of 71.43 and specificity of 96.67%. Conclusions: Our study suggests that applying the proposed score provides good interobserver agreement. The score also provided very good specificity and reasonable sensitivity in the differentiation between malignant and benign lymph nodes in the neck

2D ultrasonography and contrast enhanced ultrasound for the evaluation of cavitating mesenteric lymph node syndrome in a patient with refractory celiac disease and enteropathy T cell lymphoma

Abstract Background The cavitating mesenteric lymph node syndrome (CMLNS) is a rare manifestation of celiac disease, with an estimated mortality rate of 50%. Specific infections and malignant lymphoma may complicate its clinical course and contribute to its poor prognosis. Diagnosing the underlying cause of CMLNS can be challenging. This is the first report on contrast enhanced ultrasound (CEUS) findings in enteropathy associated T-cell lymphoma (EATL) complicating CMLNS in a gluten-free compliant patient with persistent symptoms and poor outcome. Case presentation We present the case of a 51-year old Caucasian male patient, diagnosed with celiac disease and CMLNS. Despite his compliance to the gluten-free diet the symptoms persisted and we eventually considered the possible development of malignancy. No mucosal changes suggestive of lymphoma were identified with capsule endoscopy. Low attenuation mesenteric lymphadenopathy, without enlarged small bowel segments were seen on computed tomography. CEUS revealed arterial rim enhancement around the necrotic mesenteric lymph nodes, without venous wash-out. No malignant cells were identified on laparoscopic mesenteric lymph nodes biopsies. The patient died due to fulminant liver failure 14 months later; the histopathological examination revealed CD3/CD30-positive atypical T-cell lymphocytes in the liver, mesenteric tissue, spleen, gastric wall, kidney, lung and bone marrow samples; no malignant cells were present in the small bowel samples. Conclusions CEUS findings in EATL complicating CMLNS include arterial rim enhancement of the mesenteric tissue around the cavitating lymph nodes, without venous wash-out. This vascular pattern is not suggestive for neoangiogenesis, as arteriovenous shunts from malignant tissues are responsible for rapid venous wash-out of the contrast agent. CEUS failed to provide a diagnosis in this case.</p

2D ultrasonography and contrast enhanced ultrasound for the evaluation of cavitating mesenteric lymph node syndrome in a patient with refractory celiac disease and enteropathy T cell lymphoma

BACKGROUND: The cavitating mesenteric lymph node syndrome (CMLNS) is a rare manifestation of celiac disease, with an estimated mortality rate of 50%. Specific infections and malignant lymphoma may complicate its clinical course and contribute to its poor prognosis. Diagnosing the underlying cause of CMLNS can be challenging. This is the first report on contrast enhanced ultrasound (CEUS) findings in enteropathy associated T-cell lymphoma (EATL) complicating CMLNS in a gluten-free compliant patient with persistent symptoms and poor outcome. CASE PRESENTATION: We present the case of a 51-year old Caucasian male patient, diagnosed with celiac disease and CMLNS. Despite his compliance to the gluten-free diet the symptoms persisted and we eventually considered the possible development of malignancy. No mucosal changes suggestive of lymphoma were identified with capsule endoscopy. Low attenuation mesenteric lymphadenopathy, without enlarged small bowel segments were seen on computed tomography. CEUS revealed arterial rim enhancement around the necrotic mesenteric lymph nodes, without venous wash-out. No malignant cells were identified on laparoscopic mesenteric lymph nodes biopsies. The patient died due to fulminant liver failure 14 months later; the histopathological examination revealed CD3/CD30-positive atypical T-cell lymphocytes in the liver, mesenteric tissue, spleen, gastric wall, kidney, lung and bone marrow samples; no malignant cells were present in the small bowel samples. CONCLUSIONS: CEUS findings in EATL complicating CMLNS include arterial rim enhancement of the mesenteric tissue around the cavitating lymph nodes, without venous wash-out. This vascular pattern is not suggestive for neoangiogenesis, as arteriovenous shunts from malignant tissues are responsible for rapid venous wash-out of the contrast agent. CEUS failed to provide a diagnosis in this case

Case report Ultrasound and CT imaging features in a patient with salivary duct carcinoma of the parotid gland: a case report with literature review

Abstract The aim of this paper was to present the ultrasound (US) and computed tomography (CT) appearance of a patient with salivary duct carcinoma of the parotid gland. US showed a voluminous mass of the parotid gland, with multiple calcifications. Furthermore, it revealed regional multiple lymph nodes with malignant characters. Sonoelastography of the lesion and lymph nodes detected increased rigidity. Contrast enhanced CT scan of the neck completed the data description regarding the mass expansion and invasion of surrounding tissues. US and CT imaging features played a key role in establishing the malignant character of the mass and lymph nodes

Additional file 3: Figure S3. of Dental follicle stem cells in bone regeneration on titanium implants

Alamar Blue viability assay. For testing the viability and proliferation rate of DF stem cells cultivated on titanium implants, cells seeded at a cell density of 1.2 × 105 cells/well in 12-wells plates were stained with Alamar blue solution at different periods of time (24 h, 4 and 12 days). Briefly, 100 μl of Alamar blue solution (Invitrogen) was added in each well containing 900 μl stem cell medium or differentiation medium (OS and OC). Each sample was evaluated in triplicate. After 1 h of incubation in dark at 37 °C, the medium was transferred to another 12-well plates and the absorbance was read using a BioTek Synergy 2 plate reader at 570 nm (Winooski, VT, USA). Statistical analysis was performed using t test and two-way ANOVA, Bonferroni posttest. Results: No important differences were observed between titanium implants in terms of cell viability. Statistical differences were noticed only for the 24 h culture between cell cultured on control titanium implants (Ti ctrl) and implants infiltrated with HA (Ti HA) (Figure S3). Two-way ANOVA statistical analysis revealed differences regarding the time factor (24 h vs. 12 days and 4 vs. 12 days) Figure S3: Graphical aspect of optical density values (absorbance at 570 nm) of Alamar blue staining of DF stem cells cultivated with standard stem cell medium evaluated after 24 h, 4 and 12 days (Legend: TiCtrl- Ti6Al7Nb alloy porous titanium, TiHA-titanium infiltrated with hydroxyapatite, TiSiO2-titanium infiltrated with silicatitanate). (PNG 1002 kb

Additional file 2: Figure S2. of Dental follicle stem cells in bone regeneration on titanium implants

FDA (fluorescein diacetate) viability test. DF stem cell adhesion after 1 h as well the proliferation rate during 48 h and 7 days of cultivation on titanium implants surfaces were investigated using FDA assay. Images were captured in fluorescence microscopy at 488 nm with a Zeiss Axiovert microscope. Image acquisition was performed with an AxioCam MRC camera. Figure S2: Fluorescence images captured after FDA staining of DF stemm cells after 1, 48 h and 7 days of cultivation in standard stem cell medium. (Legend: TiCtrl- Ti6Al7Nb alloy porous titanium, TiHA-titanium infiltrated with hydroxyapatite, TiSiO2-titanium infiltrated with silicatitanate) (magnification ×100). (PNG 940 kb

Additional file 5: Figure S1. of Dental follicle stem cells in bone regeneration on titanium implants

Neuronal differentiation of Df stem cells. Protocol of neuronal differentiation. DF stem cells were seeded in 6 well plates at cell density of 20 × 105 cells/well. When cells reached confluence a two steps protocol was applied: cells were cultivated for 48 h in presence of neuronal differentiation medium 1 consisting of DMEM high glucose/F12-HAM (1:1 ratio), 10 % fetal bovine serum (FBS), 1 % antibiotics,2 mM glutamine, 1 % non-essential aminoacids (NEA), supplemented with 10 ng/ml Epidermal Growth Factor (EGF), 10 ng/ml basic Fibroblast Growth Factor (bFGF), 2 % B27 and 1 % N2 Supplement. Afterwards cells were exposed to the differentiation medium 2 for 3 weeks: DMEM high glucose/F12-HAM, 10 % FBS, 1 % antibiotics, 2 mM glutamine, 1 % NEA, 1 % N2 Supplement, 2 % B27 Supplement, 3 μM all-trans retinoic acid, 0.5 mM 3-isobutyl-1-methyl-xanthine (IBMX) (all reagents were purchased from Sigma Aldrich). At the end of 3 weeks of DF stem cultivation with neuronal differentiation medium, cells were fixed and immunocytochemical stained for neurofilaments and CD 133 expression. As shown in Figure S1, the stained cells expressed positivity only for neurofilaments. Figure S1: Fluorescence image of DF stem cells induced to differentiate into neuronal cells. Cells were stained with anti neurofilaments antibody conjugated with FITC green), anti CD 133 conjugated with Texas red (red) and nuclei were counterstained with DAPI (magnification ×100). (PNG 451 kb