Materials for Controlled Release of Local Anesthetics.

Controlled release systems for prolonged duration local anesthesia have long been an area of research interest, and now are entering clinical practice, in part driven by the opioid epidemic. We discuss the design considerations and material properties of systems for controlled release of local anesthetics, from relatively simple systems to covalent binding of drugs to materials and delivery triggered by external stimuli.

Controlling Anesthesia Hardware With Simple Hand Gestures: Thumbs Up or Thumbs Down?

BACKGROUND: Modern consumer electronic devices and automobiles are often controlled by interfaces that sense physical gestures and spoken commands. In contrast, patient monitors and anesthesia devices are typically equipped with panel-mounted buttons, dials, and keyboards. The increased use of noncontact gesture-based interfaces in anesthesia may improve patient safety through more intuitive and prompter control of equipment and also through reduced rates of surface contamination. A novel gesture-based controller was designed and retrofitted to a standard GE Solar 8000M patient monitor. This type of technical innovation is rare, due to closely held proprietary input control systems on commercially produced clinical equipment. Nevertheless, we hypothesized that anesthesiologists would find a contactless gesture interface straightforward to use. METHODS: A gesture-based interface system was developed to control a Solar 8000M patient monitor using a millimeter-wave radar sensor. The system was programmed to detect noncontact "rotate" and "press" gestures to control the patient monitor by implementing a virtual trim knob for interface control. Fifty anesthesiologists tested a prototype interface and evaluated usability by completing a short questionnaire incorporating modified Likert scales. These evaluations were performed in a nonpatient care environment so that respondents were not adversely task loaded during assessment, also allaying any ethical or safety concerns regarding use of this novel interface for patient management. RESULTS: Anesthesia hardware was controlled reliably with 2 distinct gestures above the gesture sensor. The gesture-based interface generally was well received by anesthesiologists (8.09; confidence interval, 8.06-8.12 on a 10-point scale), who preferred the simpler "press" gesture to the "rotate" gesture (8.45; 8.39-8.51 vs 7.73; 7.67-7.79 on a 10-point scale; P = .005). The correlation between the preference scores for the 2 gestures from each anesthesiologist was strong (Pearson r = 0.49; 0.25-0.68; P < .001). Advancing level of training (resident, fellow, attending 1-10 years, attending >10 years) was not correlated with preference scores for either gesture (Spearman ρ = -0.02; -0.30 to 0.26; P = .87 for "press" and Spearman ρ = 0.08; -0.20 to 0.35; P = .58 for "rotate"). CONCLUSIONS: The use of gesture sensing for controlling anesthesia equipment was well received by a cohort of anesthesiologists. Even though the simpler "press" gesture was preferred over the "rotate" gesture, the intrarespondent correlation indicates that the preference for gestures as a whole is the stronger effect. No adverse relationship was found between acceptability and anesthesia experience level. Gesture sensing is a promising new area to simplify and improve the interaction between the anesthesiologist and the anesthesia workstation.

Novel DNA Aptamers that Bind to Mutant Huntingtin and Modify Its Activity.

The CAG repeat expansion that elongates the polyglutamine tract in huntingtin is the root genetic cause of Huntington's disease (HD), a debilitating neurodegenerative disorder. This seemingly slight change to the primary amino acid sequence alters the physical structure of the mutant protein and alters its activity. We have identified a set of G-quadruplex-forming DNA aptamers (MS1, MS2, MS3, MS4) that bind mutant huntingtin proximal to lysines K2932/K2934 in the C-terminal CTD-II domain. Aptamer binding to mutant huntingtin abrogated the enhanced polycomb repressive complex 2 (PRC2) stimulatory activity conferred by the expanded polyglutamine tract. In HD, but not normal, neuronal progenitor cells (NPCs), MS3 aptamer co-localized with endogenous mutant huntingtin and was associated with significantly decreased PRC2 activity. Furthermore, MS3 transfection protected HD NPCs against starvation-dependent stress with increased ATP. Therefore, DNA aptamers can preferentially target mutant huntingtin and modulate a gain of function endowed by the elongated polyglutamine segment. These mutant huntingtin binding aptamers provide novel molecular tools for delineating the effects of the HD mutation and encourage mutant huntingtin structure-based approaches to therapeutic development.

A bilayer small diameter in vitro vascular model for evaluation of drug induced vascular injury.

In pre-clinical safety studies, drug-induced vascular injury (DIVI) is defined as an adverse response to a drug characterized by degenerative and hyperplastic changes of endothelial cells and vascular smooth muscle cells. Inflammation may also be seen, along with extravasation of red blood cells into the smooth muscle layer (i.e., hemorrhage). Drugs that cause DIVI are often discontinued from development after considerable cost has occurred. An in vitro vascular model has been developed using endothelial and smooth muscle cells in co-culture across a porous membrane mimicking the internal elastic lamina. Arterial flow rates of perfusion media within the endothelial chamber of the model induce physiologic endothelial cell alignment. Pilot testing with a drug known to cause DIVI induced extravasation of red blood cells into the smooth muscle layer in all devices with no extravasation seen in control devices. This engineered vascular model offers the potential to evaluate candidate drugs for DIVI early in the discovery process. The physiologic flow within the co-culture model also makes it candidate for a wide variety of vascular biology investigations.

Comparative analysis of anti-polyglutamine Fab crystals grown on Earth and in microgravity.

Huntington's disease is one of nine neurodegenerative diseases caused by a polyglutamine (polyQ)-repeat expansion. An anti-polyQ antigen-binding fragment, MW1 Fab, was crystallized both on Earth and on the International Space Station, a microgravity environment where convection is limited. Once the crystals returned to Earth, the number, size and morphology of all crystals were recorded, and X-ray data were collected from representative crystals. The results generally agreed with previous microgravity crystallization studies. On average, microgravity-grown crystals were 20% larger than control crystals grown on Earth, and microgravity-grown crystals had a slightly improved mosaicity (decreased by 0.03°) and diffraction resolution (decreased by 0.2 Å) compared with control crystals grown on Earth. However, the highest resolution and lowest mosaicity crystals were formed on Earth, and the highest-quality crystal overall was formed on Earth after return from microgravity.

Decellularized extracellular matrix microparticles as a vehicle for cellular delivery in a model of anastomosis healing.

Extracellular matrix (ECM) materials from animal and human sources have become important materials for soft tissue repair. Microparticles of ECM materials have increased surface area and exposed binding sites compared to sheet materials. Decellularized porcine peritoneum was mechanically dissociated into 200 µm microparticles, seeded with fibroblasts and cultured in a low gravity rotating bioreactor. The cells avidly attached and maintained excellent viability on the microparticles. When the seeded microparticles were placed in a collagen gel, the cells quickly migrated off the microparticles and through the gel. Cells from seeded microparticles migrated to and across an in vitro anastomosis model, increasing the tensile strength of the model. Cell seeded microparticles of ECM material have potential for paracrine and cellular delivery therapies when delivered in a gel carrier. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1728-1735, 2016.

Anti-PolyQ Antibodies Recognize a Short PolyQ Stretch in Both Normal and Mutant Huntingtin Exon 1.

Huntington's disease is caused by expansion of a polyglutamine (polyQ) repeat in the huntingtin protein. A structural basis for the apparent transition between normal and disease-causing expanded polyQ repeats of huntingtin is unknown. The "linear lattice" model proposed random-coil structures for both normal and expanded polyQ in the preaggregation state. Consistent with this model, the affinity and stoichiometry of the anti-polyQ antibody MW1 increased with the number of glutamines. An opposing "structural toxic threshold" model proposed a conformational change above the pathogenic polyQ threshold resulting in a specific toxic conformation for expanded polyQ. Support for this model was provided by the anti-polyQ antibody 3B5H10, which was reported to specifically recognize a distinct pathologic conformation of soluble expanded polyQ. To distinguish between these models, we directly compared binding of MW1 and 3B5H10 to normal and expanded polyQ repeats within huntingtin exon 1 fusion proteins. We found similar binding characteristics for both antibodies. First, both antibodies bound to normal, as well as expanded, polyQ in huntingtin exon 1 fusion proteins. Second, an expanded polyQ tract contained multiple epitopes for fragments antigen-binding (Fabs) of both antibodies, demonstrating that 3B5H10 does not recognize a single epitope specific to expanded polyQ. Finally, small-angle X-ray scattering and dynamic light scattering revealed similar binding modes for MW1 and 3B5H10 Fab-huntingtin exon 1 complexes. Together, these results support the linear lattice model for polyQ binding proteins, suggesting that the hypothesized pathologic conformation of soluble expanded polyQ is not a valid target for drug design.

The future of medical diagnostics: large digitized databases.

The electronic health record mandate within the American Recovery and Reinvestment Act of 2009 will have a far-reaching affect on medicine. In this article, we provide an in-depth analysis of how this mandate is expected to stimulate the production of large-scale, digitized databases of patient information. There is evidence to suggest that millions of patients and the National Institutes of Health will fully support the mining of such databases to better understand the process of diagnosing patients. This data mining likely will reaffirm and quantify known risk factors for many diagnoses. This quantification may be leveraged to further develop computer-aided diagnostic tools that weigh risk factors and provide decision support for health care providers. We expect that creation of these databases will stimulate the development of computer-aided diagnostic support tools that will become an integral part of modern medicine.

Nano "fly paper" technology for the capture of circulating tumor cells.

Some efficient diagnosis and therapy systems require the isolation and quantification of circulating tumor cells (CTCs), since these species are important "biomarkers" for monitoring cancer metastasis and prognosis. Existing techniques for isolating/counting CTCs include immunomagnetic-bead-based separation and microfluidic capture. However, some of these techniques have low capture efficiency and low specificity. Through the use of a three-dimensional (3D) nanostructured substrate - specifically, a silicon-nanowire (SiNW) array coated with epithelial-cell-adhesion-molecule antibodies (anti-EpCAM) - we show that CTCs can be captured efficiently and specifically. Unlike conventional methods for isolating CTCs that depend on collision frequency and contact duration, nanoscaled local topographic interactions between the CTCs and the substrate increase their binding and markedly enhance capture efficiency.

The retention of extracellular matrix proteins and angiogenic and mitogenic cytokines in a decellularized porcine dermis.

Decellularized dermis materials demonstrate considerable utility in surgical procedures including hernia repair and breast reconstruction. A new decellularized porcine dermis material has been developed that retains many native extracellular matrix (ECM) proteins and cytokines. This material has substantial mechanical strength with maximum tensile strength of 141.7 +/- 85.4 (N/cm) and suture pull through strength of 47.0 +/- 14.0 (N). After processing, many ECM proteins remained in the material including collagen III, collagen IV, collagen VII, laminin and fibronectin. Glycosaminoglycans, including hyaluronic acid, were also preserved. Among several cytokines whose levels were quantified, more vascular endothelial growth factor (VEGF) and transforming growth factor beta (TGF-beta) were retained within this material than in comparable decellularized dermis materials. The retention of bioactivity was demonstrated in a cell culture assay. Because this decellularized porcine dermis material both retains significant strength and has substantial biological activity, it may promote rapid integration and repair in clinical applications.

Preserved extracellular matrix components and retained biological activity in decellularized porcine mesothelium.

Mesothelium tissues such as peritoneum and pleura have a thin and strong layer of extracellular matrix that supports mesothelial cells capable of rapid healing. Decellularized porcine mesothelium was characterized for strength, composition of the matrix and biological activity. The tensile strength of the material was 40.65 +/- 21.65 N/cm. Extracellular matrix proteins collagen IV, fibronectin, and laminin as well as glycosaminoglycans were present in the material. Cytokines inherent in the extracellular matrix were preserved. Vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF) and transforming growth factor beta (TGF-beta) were retained and the levels of VEGF and TGF-beta in the decellularized mesothelium were higher than those found in decellularized small intestinal submucosa (SIS). The decellularized mesothelium also stimulated human fibroblasts to produce more VEGF than fibroblasts grown on tissue culture plastic. Decellularized mesothelium is a sheet material with a combination of strength and biological activity that may have many potential applications in surgical repair and regenerative medicine.