Cytotoxicity and bioactive potential of new root repair materials for use with BMP-2 transfected human osteoblast cells

Abstract: Modified formulations of calcium silicate repair materials with additives have been developed to enhance handling, consistency, biocompatibility and bioactivity. Considering the relevance of osteoblastic cell response to mineralized tissue repair, human osteoblastic cells (Saos-2 cells overexpressing BMP-2) were exposed to mineral trioxide aggregate (MTA) (with calcium tungstate - CaWO4), MTA HP Repair, Bio-C Repair and Bio-C Pulpo. Cell viability was assessed by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) and neutral red (NR), and cell death, by flow cytometry. Gene expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX-2), and alkaline phosphatase (ALP) osteogenic markers were evaluated by real-time polymerase chain reaction (RT-qPCR). ALP activity and alizarin red staining (ARS) were used to detect mineralization nodule deposition. Bioactive cements presented no cytotoxic effect, and did not induce apoptosis at the higher dilution (1:12). MTA, Bio-C Repair and Bio-C Pulpo exhibited higher ALP activity than the control group (P < 0.05) after 7 days. MTA, MTA HP and Bio-C Pulpo affected the formation of mineralized nodules (p < 0.05). Exposure to all cement extracts for 1 day increased BMP-2 gene expression. RUNX-2 mRNA was greater in MTA, MTA HP and Bio-C Repair. MTA, MTA HP and Bio-C Pulpo increased the ALP mRNA expression, compared with BMP-2 unexposed cells (P < 0.05). Calcium silicate cements showed osteogenic potential and biocompatibility in Saos-2 cells transfected BMP-2, and increased the mRNA expression of BMP-2, RUNX-2, and ALP osteogenic markers in the BMP-2 transfected system, thereby promoting a cellular response to undertake the mineralized tissue repair.

Perillyl alcohol has antibacterial effects and reduces ROS production in macrophages

Abstract Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 μM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 μM (p<0.05) and 250 μM (p<0.01). The POH increased ROS production at both 10 μM and 100 μM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 μM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 μM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.

Evaluation of previous abdominal pain episodes in women with biliary pancreatitis: possible higher incidence of recurrent attacks.

Background and aim: Recurrent biliary pancreatitis is described as episodes of new abdominal pain after diagnosis of pancreatitis. Few studies have analyzed the abdominal pain before the diagnosis of acute pancreatitis. Our study aimed to analyze factors associated with previous abdominal pain episodes in patients with biliary pancreatitis, and elucidate its possible pancreatic origin. Methods: Data from direct interrogation and medical records was analyzed from 48 hospitalized female patients with diagnosis of acute biliary pancreatitis. Results: Mean age of our patients was 31.6 years (SD+13.9). Forty one (85.4%) patients gave history of at least one previous abdominal pain episode. During the episode 37 (90.2%) patients received H2 receptor antagonist or proton pump inhibitors as treatment; 26 (63.4%) had epigastric pain; 23(56.1%) gave association with cholecystokinetic food; 21 (51.2%) complained of nausea and/or vomiting; 23 (56.1%) had jaundice, acholia and/or dark urine; and 20 (48.9%) patients had microlithiasis and/or biliary sludge. Conclusions: Previous abdominal pain episodes had similar characteristics to a pancreatic episode in a high percentage of our patients. These characteristics suggest that these episodes are often undiagnosed pancreatic attacks.

Eficacia del Enalapril en pacientes monorrenos con dieta normoproteica y sin ella / Efficacy of Enalapril in patients with single kidney and with or without normoprotein diet

El paciente monorreno, sin dieta normoproteica,puede presentar microalbuminuriaanormal con eventual desarrollo de glomerulosclerosis.Objetivo. Evaluar la eficacia del Enalapril parareducir la microalbuminuria patológica en pacientesmonorrenos con dieta normoproteica y sin ella.Población, material y métodos. Estudiamos 49 pacientesmonorrenos, edad (media) 11 años, eutróficos,con función renal normal y consentimiento médicoaprobado. Criterios de exclusión: uropatías,hipertensión arterial, enfermedades oncológicas, tratamientoscon drogas nefrotóxicas, proteinuriaortostática. La población se clasificó en dos grupos:Grupo A: con dieta normoproteica (0,8-1,2 g/kg/díapara sexo y edad). Grupo B: con dieta libre.A los pacientes con microalbuminuria anormal seles administró Enalapril (intervalo de dosis: 0,1 mg/kg/día a 0,3 mg/kg/día).Resultados. Grupo A: 21 pacientes, 2 con microalbuminuriapatológica. Aumento promedio del tamañorenal: 14% (DE 8 por ciento). Riesgo de presentarmicroalbuminuria patológica: 9 porc ciento.Grupo B: 28 pacientes, 11 con microalbuminuriaanormal. Riesgo de presentar microalbuminuriaanormal: 40 por ciento. Aumento promedio del tamaño renal:33,8 (DE 6,1 por ciento). RR: 4,125(IC 1-16) (p: 0,01).NND: 3. Media de microalbuminuria pre-Enalapril50,20 μg/minuto y post-Enalapril 11,5 μg/minuto.Dosis media de Enalapril: 0,2 mg/kg/día.Coeficiente de correlación entre microalbuminuriay aumento del tamaño renal: 0,75; y entre ingestaproteica y aumento del tamaño renal: 0,72.Tiempo (medio) de aparición de la microalbuminuriaanormal: 81 meses (DE 7 meses).Conclusión. El Enalapril junto con la dieta normoproteicaen pacientes monorrenos disminuyen elriesgo de desarrollar microalbuminuria anormal