RESUMEN
An in vitro model for neural trauma was characterized and validated. The model is based on a novel device that is capable of applying high strain rate, homogeneous, and equibiaxial deformation to neural cells in culture. The deformation waveform is fully arbitrary and controlled via closed-loop feedback. Intracellular calcium ([Ca2+]i) alterations were recorded in real time throughout the imposed strain with an epifluorescent microscopy system. Peak change in [Ca2+]i recovery of [Ca2+]i and percent responding NG108-15 cells were shown to be dependent on strain rate (1(-1) to 10(-1)) and magnitude (0.1 to 0.3 Green's Strain). These measures were also shown to depend significantly on the interaction between strain rate and magnitude. This model for neural trauma is a robust system that can be used to investigate the cellular tolerance and response to traumatic brain injury.
Asunto(s)
Calcio/fisiología , Modelos Biológicos , Traumatismos del Sistema Nervioso/fisiopatología , Análisis de Varianza , Fenómenos Biomecánicos , Calcio/metabolismo , Homeostasis , Traumatismos del Sistema Nervioso/metabolismoRESUMEN
The strength of adhesion of NG108-15 cells to glass substrates modified with adsorbed proteins (laminin and poly-ornithine) or modified with covalently bound peptides (tri-ornithine and Tyr-Ile-Gly-Ser-Arg) was quantitatively assessed, by determining the shear stresses necessary to denude the cells from substrates using a spinning disk device. The shear stresses required to detach NG108-15 cells from glass modified with either adsorbed poly-ornithine or with both poly-ornithine and laminin were significantly (P < 0.05) higher than the shear stresses required to detach the cells from plain glass substrates. Covalent surface modifications resulted in higher strengths of NG108-15 adhesion than were exhibited on surfaces modified with adsorbed proteins. NG108-15 cell adhesion strength was maximal on surfaces covalently modified with only amine groups (without any peptides or proteins). These results indicate that general (i.e., not necessarily receptor-specific) surface modification strategies, which increase the net surface charge of a substrate, will elicit strong adhesion of NG108-15 cells.
Asunto(s)
Moléculas de Adhesión Celular/fisiología , Adhesión Celular , Neuronas/citología , Técnicas de Cultivo de Célula , Humanos , Laminina/fisiología , Microscopía de Contraste de Fase , Neuronas/metabolismo , Péptidos/fisiología , Estrés Mecánico , Propiedades de Superficie , Células Tumorales CultivadasRESUMEN
PURPOSE: Others have shown that the fetal bovine bladder is relatively noncompliant. Previous studies on compliance of fetal bovine bladders have demonstrated that the youngest fetal bladders had lowest and the oldest fetal bladders (near full-term) had greatest compliance. Our study was designed to determine the level of participation of active tension in the compliance of fetal bladders during gestation. MATERIALS AND METHODS: Fetal bovine bladders were obtained immediately after maternal harvest and crown-to-rump length was measured to determine gestational age. The fetus was inspected for genitourinary anomalies and the bladder was immediately placed in chilled M199 media. Strips (1 x 0.5 cm.) were excised from the anterior sagittal plane of the bladder and subjected to length-tension analysis in oxygenated Tyrode's buffer at 37C. Tension was measured using a force transducer and length was increased using a micropositioner. Compliance refers to the length-tension studies performed in normal Tyrode's solution and consists of a combination of active (smooth muscle tone) and passive properties. Passive compliance refers to length-tension studies performed after inactivation of bladder smooth muscle tone. Compliance with muscle tone intact was determined by incrementally stretching the strips to twice resting length in physiological buffer and then permitting them to return to resting length. Passive compliance with muscle tone ablated was determined in the same fashion after overnight incubation in calcium-free Tyrode's buffer in the presence of 5 mM. egtazic acid and 10 mM. sodium azide. An exponential function was fit to the normalized length-tension curves, where the exponential coefficient (EC) is numerically inversely proportional to compliance. RESULTS: Passive compliance was greatest in the youngest bladders (EC = 0.5 in the first trimester) and gradually decreased with increasing fetal age (EC = 1.2 in the third trimester). Active compliance demonstrated the opposite pattern, since the younger bladders were more stiff (EC = 2.1 in the first and 1.6 in the third trimesters). CONCLUSIONS: These studies demonstrate that passive compliance is greatest in the youngest bladders and progressively decreases with gestation. However, active smooth muscle tone is greatest in the youngest bladders and decreases with gestation. Thus, high active smooth muscle tone in the youngest fetal bladders results in relatively poor compliance of the early stage fetal bladder.
Asunto(s)
Vejiga Urinaria/embriología , Vejiga Urinaria/fisiología , Animales , Bovinos , Edad Gestacional , Músculo Liso/fisiologíaRESUMEN
Shortening of the first metatarsal has been one of the reported complications noted in association with base osteotomies. Previous studies have used radiographs as a means of assessing metatarsal length, the reliability of which has been subject to question. In this study, the authors have determined the amount of shortening that may be anticipated with a closing base wedge osteotomy of the first metatarsal through an experimental model. A mathematical model was also devised that confirmed these findings. Additional studies were performed to determine what influences other variables may have on this procedure.
Asunto(s)
Hallux Valgus/cirugía , Metatarso/cirugía , Osteotomía/métodos , Humanos , Modelos Teóricos , Resultado del TratamientoRESUMEN
The short-term (less than 2 min) alterations in the intracellular free calcium concentration in differentiated NG108-15 (neuroblastoma cross glioma) cells exposed to dynamic mechanical deformation with and without superimposed chemical hypoxia were determined. A previously developed device, modified for these studies, was used to apply deformations at a magnitude and rate representative of those experienced by neural tissue in Traumatic Brain Injury. Chemical hypoxia was imposed using a combination of 2-deoxy-D-glucose and salicylate, anaerobic and aerobic metabolic blockers, respectively. Real time measurement of intracellular free calcium concentration using Fura-2 and a custom epifluorescence microscopy system provided a quantitative index of cell response. At high rates of deformation (approximately 10 sec-1), increases in intracellular free calcium concentration were exponentially related to the magnitude of the applied deformation. Chemical hypoxia had no effect on this acute response. At low rates of deformation, small increases in intracellular free calcium concentration were independent of the magnitude of the deformation. These findings indicate that strategies for reducing severity of TBI should focus on minimizing the rate of deformation of neural cells. Together with data from animal, physical, and finite element models, these data can be employed in the development of physiologic injury tolerance criteria for the whole head.
Asunto(s)
Lesiones Encefálicas/metabolismo , Calcio/metabolismo , Hipoxia/metabolismo , Heridas y Lesiones/metabolismo , Animales , Modelos Animales de Enfermedad , Técnicas In VitroRESUMEN
Foot ulcerations are one of the most common and dangerous complications associated with chronic diabetes mellitus. Many studies have focused on neuropathy, in conjunction with elevated ground reactive forces, as the principal cause of these ulcerations. The authors discuss the mechanical cause of diabetic ulcerations at the cellular level. It is hypothesized that increased rate of tissue deformation associated with foot slap secondary to progressive motor neuropathy is the actual culprit, and not the magnitude of local pressure applied. The authors present a cellular model that shows that high rates of tissue deformation may result in elevated intracellular calcium concentrations, which may lead to cellular death, while comparable loads gradually applied do not. Furthermore, there is no significant difference in the response observed at 5 psi and 10 psi. Based on these findings, it is hypothesized that techniques such as ankle foot orthoses, which control the velocity of foot strike, may be useful in treating diabetic foot ulcerations.