Limited beneficial effects of perfluorocarbon emulsions on encapsulated cells in culture: experimental and modeling studies.

Due to the high solubility of oxygen in perfluorocarbons (PFCs), these compounds have been explored for improved cell and tissue oxygenation. The goal of this study is to investigate the effects of a PFC emulsion on cellular growth and function in a tissue engineered construct. A perfluorotributylamine (PFTBA) emulsion was co-encapsulated at 10 vol% with mouse ßTC-tet insulinoma cells in calcium alginate beads and cultured under normoxic and severely hypoxic conditions. The number of metabolically active cells and the induced insulin secretion rate were measured over time for up to 16 days. Results showed no significant effect of PFTBA relative to the PFTBA-free control. The alginate-PFC-cell system was also modeled mathematically, and simulations tracked the number of viable cells over time under the same conditions used experimentally. Simulations revealed only a small, likely experimentally undetectable difference in cell density between the PFC-containing and PFC-free control beads. It is concluded that PFTBA up to 10 vol% has no significant effect on the growth and function of encapsulated ßTC-tet cells under normoxic and hypoxic conditions.

Monitoring of dissolved oxygen and cellular bioenergetics within a pancreatic substitute.

This work investigated the use of nuclear magnetic resonance (NMR) spectroscopy in combination with a mathematical model of an encapsulated cell system as a method for rapidly assessing the status of a pancreatic substitute. To validate this method, an in vitro experiment was performed in which the encapsulated cells were perfused in an NMR-compatible system and the dissolved oxygen (DO) concentration of the perfusing medium was lowered from 0.20 to 0.05 mM, then returned to 0.20 mM in a stepwise fashion. The cellular metabolic activity and bioenergetics were evaluated by measuring the oxygen consumption rate (via DO sensors) and nucleotide triphosphate levels (via (31)P NMR). By incorporating a perfluorocarbon emulsion into the alginate beads, the cellular oxygenation state was monitored by measuring the average intrabead DO (AIDO) concentration by (19)F NMR. The in vitro measurements were then compared with model predictions based on the measured external DO concentration and time. Model-predicted cell growth and AIDO closely matched the experimentally acquired data. As the DO concentrations both external to and within the pancreatic substitute are needed to apply this methodology in vivo, the feasibility of measuring the DO concentration from two distinct bead populations implanted in the peritoneal cavity of mice was established. It is concluded that PFC incorporation and (19)F NMR measurements, in combination with a mechanistic model of the encapsulated system, allow the tracking of the state of a pancreatic substitute in vitro and potentially in vivo.

Modeling of encapsulated cell systems.

Tissue engineered substitutes consisting of cells in biocompatible materials undergo remodeling with time as a result of cell growth and death processes. With inert matrices that do not directly influence cell growth, remodeling is driven mainly by the concentration of dissolved oxygen (DO). Insulin-secreting cell lines encapsulated in alginate-based beads and used as a pancreatic substitute represent such a case. Beads undergo remodeling with time so that an initially homogeneous distribution of cells is eventually replaced by a dense peripheral ring of primarily viable cells, whereas inner cells are mostly necrotic. This paper develops and analyzes a mathematical model of an encapsulated cell system of spherical geometry that tracks the viable and dead cell densities and the concentration of DO within the construct as functions of radial position and time. Model simulations are compared with experimental histology data on cell distribution. Correlations are then developed between the average intrabead DO concentration (AIDO) and the total viable cell number, as well as between AIDO and the radial cell and DO distributions in beads. As AIDO can be measured experimentally by incorporating a perfluorocarbon emulsion in the beads and acquiring (19)F nuclear magnetic resonance (NMR) spectroscopic data, these correlations can be used to track the remodeling that occurs in the construct in vitro and potentially in vivo. The usefulness of mathematical models in describing the dynamic changes that occur in tissue constructs with time, and the value of these models at obtaining additional information on the system when used interactively with experimental measurements, are discussed.

Mini-surgical approach for spinal endoscopy in the presence of stenosis of the sacral hiatus.

Spinal endoscopy is a useful tool for the management of intractable low back or radicular pain originating from post lumbar laminectomy syndrome, epidural scarring, or disc protrusions, and non-responsive to conservative modalities and other interventional techniques including fluoroscopically directed epidural steroid injections and percutaneous adhesiolysis. Spinal endoscopy requires that the caudal canal be entered via the sacral hiatus. However, in a very small proportion of patients, access to the caudal canal is restricted because of stenosis or cartilaginous overgrowth of the hiatus. In such cases, the procedure is stopped because of the absence of an alternative approach to enter the epidural space with the spinal endoscope, resulting in non-availability of this treatment. This report describes a novel method of dealing with the problem of cartilaginous obstruction of the sacral hiatus, using a mini-surgical approach to decompress the hiatus, allowing access into the caudal canal.

Comparison of registration accuracy of skin- and bone-implanted fiducials for frameless stereotaxis of the brain: a prospective study.

The registration accuracy of skin- and bone-implanted fiducials using a frameless stereotactic system were analyzed prospectively.Twenty-eight patients underwent resection of intra-axial neoplasmas after both skin- and bone-implantable fiducial markers were placed. Both sets of fiducials were independently co-registered to a magnetic resonance imaging data set acquired preoperatively using the ISG Viewing Wandtrade mark. Root mean square errors were recorded as an objective measure of registration accuracy of the two types of fiducials.Root mean square errors of bone-implanted fiducials registration were lower than those of skin fiducials; however, this difference was not statistically significant (p = 0.206).The registration accuracy of skin- and bone-implanted fiducials appears to be similar. Still, bone-implanted fiducials may be advantageous compared to skin fiducials when re-registration of the patient-image space is desired intraoperatively such as during major drift in the patient's position or after surgical repositioning.