Cytotoxic and anti-inflammatory effects of chitosan and hemostatic gelatin in oral cell culture.

Chitosan is a biopolymer with bactericidal/bacteriostatic effect, biocompatible and biodegradable. It has been used in tissue engineering to replace tissues partially or completely by releasing bioactive materials or influencing cell growth, usually in regenerative medicine and dentistry. The aim of this study was to evaluate the cytotoxic and anti-inflammatory effect of chitosan alone or with hemostatic gelatin (Spongostand®) in cultures of human pulp cells (HPC), human gingival fibroblasts (HGF) and mouse pre-osteoblasts (MC3T3-E1, ATCC). HPC and HGF were isolated from patients. Cells were subcultured in DMEM. Chitosan was inoculated at different concentrations (0-0.5%) and hemostatic gelatins impregnated with chitosan (0.19%) were placed directly in the presence of cells and incubated for 24 hours. Cell viability was determined by MTT method and mean cytotoxic concentration (CC50) was calculated from the dose-response curve. Anti-inflammatory effect was calculated from the in vitro gingivitis model induced with interleukin 1beta (IL-1ß) in HGF and protein detection. The data were subjected to Shapiro-Wilk, Kruskal-Wallis and Mann-Whitney tests. Experiments were performed in triplicate of three independent assays. Cell viability of HPC, HGF and MC3T3-E1 in contact with chitosan decreased significantly (p<0.05). The HPC were the most sensitive (CC50= 0.18%), followed by HGF (CC50= 0.18%) and MC3T3-E1 (CC50= 0.19%). The cytotoxicity of gelatins impregnated with chitosan decreased cell viability of HGF and HPC by 11% and 5%, respectively. The proinflammatory effect was reduced significantly in the gingivitis model. To conclude, chitosan induces moderate cytotoxic effects alone or with hemostatic gelatin at 0.19%, in dose-dependent manner, with anti-inflammatory effects on human gingival fibroblasts. The use of chitosan as a biomaterial can be an excellent choice for use in regenerative dentistry.

El quitosano es un biopolímero con efecto bactericida/bacteriostático, biocompatible y biodegradable. Se ha utilizado en ingeniería de tejidos con el fin de reemplazar parcial o completamente los tejidos como material bioactivo o influyendo en el crecimiento celular, comúnmente, para medicina y odontología regenerativa. Evaluar el efecto citotóxico y antiinflamatorio del quitosano solo o con gelatina hemostática (Spongostand®) en cultivos con células pulpares humanas (HPC), fibroblastos gingivales humanos (HGF) y preosteoblastos de ratón (MC3T3-E1, ATCC). HPC, HGF se aislaron de pacientes. Las células se subcultivaron en DMEM. Se inoculó quitosano a diferentes concentraciones (0-0,5%) y se colocaron gelatinas hemostáticas impregnadas con quitosano (0,19%) directamente en presencia de células y se incubaron durante 24 horas. La viabilidad celular se determinó mediante el método MTT y se calculó la concentración citotóxica media (CC50) a partir de la curva dosis-respuesta. El efecto antiinflamatorio se calculó a partir del modelo de gingivitis in vitro inducido con interleucina 1ß (IL-1ß) en HGF. Los datos se sometieron a las pruebas de Shapiro-Wilk, Kruskal-Wallis y Mann-Whitney. Los experimentos se realizaron por triplicado de tres ensayos independientes. La viabilidad celular de HPC, HGF y MC3T3-E1 en contacto con el quitosano disminuyó significativamente la viabilidad celular (p<0.05). Las HPC fueron las más sensibles (CC50= 0,18%) seguido de HGF (CC50= 0,18%) y MC3T3-E1 (CC50= 0,19%). Las gelatinas impregnadas con quitosano mostraron una disminución en la viabilidad celular para HGF, HPC de 11% y 5% respectivamente y se redujo significativamente el efecto pro-inflamatorio en el modelo de gingivitis humano. El quitosano induce efectos citotóxicos moderados solo o con gelatina hemostática a 0,19% de forma dosis-dependiente con efectos antiinflamatorios en fibroblastos gingivales humanos. El uso de quitosano como biomaterial puede ser una excelente opción para su uso en odontología regenerativa.

Characterizing and Tuning Perfusion Parameters Within an Innovative, Versatile Oxygenating Perfusion System.

The advantages of oxygenated perfusion are continuing to be demonstrated by many groups focused on improving the efficacy of tissue preservation for transplant, bioreactors for studies of basic tissue physiology, and closed-loop resuscitation. This work presents a novel and portable device that supplies oxygenated and pulsatile perfusion, both of which are regulated by a single pump-oxygenator component comprised of silicone tubes that are cyclically inflated/deflated with compressed oxygen. In this study, pump variables (oxygen supply pressure and length of a silicone tube) were evaluated against hydraulic elements that mimicked the vascular resistance of kidneys, livers, and hearts. The perfusion pressures, flow rates, and oxygenation rates produced by the device were characterized for all configurations of pump variables, and the pulse rates were tuned to improve performance. The device supplied perfusion pressures ranging from 3.5 to 109 mmHg, flow rates ranging from 1.4 to 71.8 mL min-1, and oxygenation rates up to 316.6 µmol min-1. From those results, it was determined that the device was capable of achieving perfusion parameters used in previous kidney, liver, and heart preservation studies. Ultimately, this research demonstrated the efficacy of a novel device that is designed to supply oxygenated perfusion across a range of applications.

Cytotoxic and anti-inflammatory effects of chitosan and hemostatic gelatin in oral cell culture / Efectos citotóxicos y antiinflamatorios del quitosano y la gelatina hemostática en cultivo celular oral

ABSTRACT Chitosan is a biopolymer with bactericidal/bacteriostatic effect, biocompatible and biodegradable. It has been used in tissue engineering to replace tissues partially or completely by releasing bioactive materials or influencing cell growth, usually in regenerative medicine and dentistry. The aim of this study was to evaluate the cytotoxic and anti-inflammatory effect of chitosan alone or with hemostatic gelatin (Spongostand®) in cultures of human pulp cells (HPC), human gingival fibroblasts (HGF) and mouse pre-osteoblasts (MC3T3-E1, ATCC). HPC and HGF were isolated from patients. Cells were subcultured in DMEM. Chitosan was inoculated at different concentrations (0-0.5%) and hemostatic gelatins impregnated with chitosan (0.19%) were placed directly in the presence of cells and incubated for 24 hours. Cell viability was determined by MTT method and mean cytotoxic concentration (CC50) was calculated from the dose-response curve. Anti-inflammatory effect was calculated from the in vitro gingivitis model induced with interleukin 1beta (IL-1β) in HGF and protein detection. The data were subjected to Shapiro-Wilk, Kruskal-Wallis and Mann-Whitney tests. Experiments were performed in triplicate of three independent assays. Cell viability of HPC, HGF and MC3T3-E1 in contact with chitosan decreased significantly (p<0.05). The HPC were the most sensitive (CC50= 0.18%), followed by HGF (CC50= 0.18%) and MC3T3-E1 (CC50= 0.19%). The cytotoxicity of gelatins impregnated with chitosan decreased cell viability of HGF and HPC by 11% and 5%, respectively. The proinflammatory effect was reduced significantly in the gingivitis model. To conclude, chitosan induces moderate cytotoxic effects alone or with hemostatic gelatin at 0.19%, in dose-dependent manner, with anti-inflammatory effects on human gingival fibroblasts. The use of chitosan as a biomaterial can be an excellent choice for use in regenerative dentistry.

RESUMEN El quitosano es un biopolímero con efecto bactericida/bacteriostático, biocompatible y biodegradable. Se ha utilizado en ingeniería de tejidos con el fin de reemplazar parcial o completamente los tejidos como material bioactivo o influyendo en el crecimiento celular, comúnmente, para medicina y odontología regenerativa. Evaluar el efecto citotóxico y antiinflamatorio del quitosano solo o con gelatina hemostática (Spongostand®) en cultivos con células pulpares humanas (HPC), fibroblastos gingivales humanos (HGF) y preosteoblastos de ratón (MC3T3-E1, ATCC). HPC, HGF se aislaron de pacientes. Las células se subcultivaron en DMEM. Se inoculó quitosano a diferentes concentraciones (0-0,5%) y se colocaron gelatinas hemostáticas impregnadas con quitosano (0,19%) directamente en presencia de células y se incubaron durante 24 horas. La viabilidad celular se determinó mediante el método MTT y se calculó la concentración citotóxica media (CC50) a partir de la curva dosis-respuesta. El efecto antiinflamatorio se calculó a partir del modelo de gingivitis in vitro inducido con interleucina 1β (IL-1β) en HGF. Los datos se sometieron a las pruebas de Shapiro-Wilk, Kruskal-Wallis y Mann-Whitney. Los experimentos se realizaron por triplicado de tres ensayos independientes. La viabilidad celular de HPC, HGF y MC3T3-E1 en contacto con el quitosano disminuyó significativamente la viabilidad celular (p <0.05). Las HPC fueron las más sensibles (CC50= 0,18%) seguido de HGF (CC50= 0,18%) y MC3T3-E1 (CC50= 0,19%). Las gelatinas impregnadas con quitosano mostraron una disminución en la viabilidad celular para HGF, HPC de 11% y 5% respectivamente y se redujo significativamente el efecto pro-inflamatorio en el modelo de gingivitis humano. El quitosano induce efectos citotóxicos moderados solo o con gelatina hemostática a 0,19% de forma dosis-dependiente con efectos antiinflamatorios en fibroblastos gingivales humanos. El uso de quitosano como biomaterial puede ser una excelente opción para su uso en odontología regenerativa.

Análisis de Patrones de Cierre Velofaríngeo en Pacientes con Labio y Paladar Hendido / Pattern Analysis of Velopharyngeal Closure in Patients With Cleft Lip and Palate

El objetivo fue identificar los patrones de Cierre Velofaríngeo (CVF) en pacientes con Paladar Hendido (PH) y Labio y Paladar Hendido (LPH) que acuden a la Clínica de Atención Integral de Pacientes con Labio y Paladar Hendido (CAIPLPH) de la Escuela Nacional de Estudios Superiores, Unidad León (ENES, León). Estudio transversal, con una muestra por conveniencia de 79 pacientes entre 4 y 10 años de edad. Las variables incluidas fueron el patrón de CVF, tipo de hendidura, edad del paciente y el antecedente de intervención quirúrgica previa para el cierre de la hendidura palatina. A los pacientes se les realizó valoración clínica y nasofibroscopía por examinadores previamente estandarizados para conocer si existe una relación entre edad y patrón de CVF, entre el tipo de hendidura y patrón de CVF y una relación entre el patrón de CVF y el antecedente quirúrgico, utilizando una prueba bivariada de Chi cuadrada. Se examinaron 46 hombres y 33 mujeres con una media de edad 6,6 años. La frecuencia de los patrones de CVF fueron coronal 8,86%, sagital 67,08%, circular 13,92% y circular con rodete de Passavant 10,12%. Se observó una diferencia estadística significativa entre el patrón de CVF y el tipo de hendidura (X2= 53,93, p<0,001). Las diferentes formas en que se pueden presentar las hendiduras de paladar y labiopalatinas influyen en la función del CVF ya que se puede presentar un patrón distinto en cada caso. El patrón de CVF no se ve afectado por la edad ni por el antecedente quirúrgico de cierre de la hendidura palatina.

The objective of the study was to identify patterns of velopharyngeal closure (CVF) in patients with Cleft Palate (PH) and cleft lip and palate (LPH) attending the Clinic of Comprehensive Attention of Patients with Cleft Lip and Palate (CAIPLPH) of the Escuela Nacional de Estudios Superiores, Unidad León (ENES, León). A cross-sectional study and convenience sample of 79 patients was carried out, including those aged between 4 and 10 years. The variables included were: CVF pattern, kind of cleft, the patient's age and history of previous surgery for closure of the palatal cleft. Patients underwent clinical evaluation and nasofibroscopy by examiners previously standardized to know if there is a relationship between age and CVF pattern, between kind of cleft and pattern of CVF and a relationship between the pattern of CVF and patient's history of previous surgery, using a X2 test bivariate square. Forty-six men and 33 women with a mean age 6.6 years were examined. Frequency patterns CVF: 8.86% coronal, sagittal 67.08%, circular 13.92% and a circular with Passavant ridge 10.12%. A statistically significant difference between the pattern of CVF and the kind of cleft (X2= 53.93, p <0.001) was observed. The different ways in which the PH and LPH clefts can present influence the function of CVF as it can present a different pattern in each case. CVF pattern is unaffected by age or by the previous surgical closure.

Alterations and pathologies prevalence in panoramic radiographs in patients attending the university dental clinic / Prevalencia de alteraciones y patologías en radiografías panorámicas en pacientes atendidos en una clínica dental universitaria

The aim was to report the prevalence of alterations and pathologies of the analysis of digital panoramic radiographs in the clinic of the Dental School at Escuela Nacional de Estudios Superiores, Unidad León, since October 2011 until April 2012. This was developed as a retrospective study involving 516 digital panoramic radiographs upon which was performed a visual analysis to report the prevalence of abnormalities or pathologies. The findings were related to maxillary sinus abnormality or pathology, nasal cavity, maxillary and mandibular bones and dental abnormalities. The panoramic radiograph because of its versatility is still one of the most important oral and maxillofacial diagnosis methods, it helps us to detect pathologies, which do not always have symptomatology and could cause health problems.

El objetivo fue describir la prevalencia de alteraciones y patologías en radiografías panorámicas digitales de la clínica de la Facultad de Odontología de la Escuela Nacional de Estudios Superiores, Unidad León, desde octubre de 2011 hasta abril de 2012. Se desarrolló un estudio retrospectivo que incluyó a 516 radiografías panorámicas digitales en las que se realizó un análisis visual para informar de la prevalencia de anomalías o patologías. Los hallazgos se relacionaron con anormalidad o patología del seno maxilar, cavidad nasal, hueso maxilar y mandibular, y alteraciones dentales. La radiografía panorámica debido a su versatilidad sigue siendo uno de los métodos de diagnóstico oral y maxilofacial más importantes, que nos ayuda a detectar patologías, que no siempre tienen la sintomatología y podrían causar problemas de salud.