In-vivo Intradermal Delivery of Co-57 labeled Vitamin B-12, and Subsequent Comparison with Standard Subcutaneous Administration.

Vitamin B-12 (cobalamin) deficiency in humans is a worldwide problem emanating from varied causes such as insufficient dietary intake or malabsorption of the micronutrient due to an underlying condition (absence or failure of intrinsic factor, atrophic gastritis, post-operative bariatric surgery, inflammatory bowel disease, cobalt deficiency etc.). As oral supplementation is limited by its bioavailability due to the absorptive property of intrinsic factor, clinicians often prescribe parenteral forms of administration to replenish diminished levels rapidly. The gold standard in parenteral delivery of cobalamin is subcutaneous and/or intramuscular injections. The relatively large molecular size of cobalamin (1355.39 Da) makes passive transdermal patch-based delivery via the stratum corneum quite challenging. Hence, the primary goal of this study is to investigate the feasibility of intradermal (ID) delivery of Vitamin B-12 via an almost painless microneedle injection and subsequent comparison with standard subcutaneous (SC) delivery. This work reports on a custom-made microneedle device built from a commercial insulin needle and it's use to perform ID delivery of Co-57 radiolabeled Vitamin B-12 in-vivo in rabbits. The pharmacokinetic profile and bioavailability were studied and compared with SC delivery. It is the first comprehensive study, to our best knowledge, that compares a micronutrient (eg. Vitamin B-12) delivery via ID and SC routes in-vivo. While the bioavailability for the SC route is found to be slightly higher compared to the ID route (99% vs. 96%), the Tmax for both are almost identical. Thus, ID delivery of Vitamin B-12 using a microneedle injection could be a viable and minimally invasive alternative to existing parenteral options.

Digital microfluidics for time-resolved cytotoxicity studies on single non-adherent yeast cells.

Single cell analysis (SCA) has gained increased popularity for elucidating cellular heterogeneity at genomic, proteomic and cellular levels. Flow cytometry is considered as one of the most widely used techniques to characterize single cell responses; however, its inability to analyse cells with spatio-temporal resolution poses a major drawback. Here, we introduce a digital microfluidic (DMF) platform as a useful tool for conducting studies on isolated yeast cells in a high-throughput fashion. The reported system exhibits (i) a microwell array for trapping single non-adherent cells by shuttling a cell-containing droplet over the array, and allows (ii) implementation of high-throughput cytotoxicity assays with enhanced spatio-temporal resolution. The system was tested for five different concentrations of the antifungal drug Amphotericin B, and the cell responses were monitored over time by time lapse fluorescence microscopy. The DMF platform was validated by bulk experiments, which mimicked the DMF experimental design. A correlation analysis revealed that the results obtained on the DMF platform are not significantly different from those obtained in bulk; hence, the DMF platform can be used as a tool to perform SCA on non-adherent cells, with spatio-temporal resolution. In addition, no external forces, other than the physical forces generated by moving the droplet, were used to capture single cells, thereby avoiding cell damage. As such, the information on cellular behaviour during treatment could be obtained for every single cell over time making this platform noteworthy in the field of SCA.

A novel method for monolithic fabrication of polymer microneedles on a platform for transdermal drug delivery.

This paper reports on the creation of a novel method for monolithic fabrication of out-of-plane polymer (SU-8) microneedles incorporating sharpness of needle-tips, hollowness of needle lumen as well as a platform on which the microneedles stand orthogonally with the hollow of the needle lumen continuous through the platform. In essence, both the microneedle as well as the platform on which it stands, are made of the same polymer material, rendering the process monolithic. The microneedle tips produced were quite sharp with tip diameters ranging between 5 to 10 µm, needle heights greater than 1 mm and resulting aspect ratio of 40. Further, mechanical tests performed on the fabricated microneedles demonstrate a critical compressive failure load of about 173 mN on average per microneedle, which translates into a safety factor greater than one for skin penetration.

Out-of-plane, high strength, polymer microneedles for transdermal drug delivery.

This paper reports on the high strength of high-aspect ratio (> 50) hollow, polymer microneedles fabricated out-of-plane using a fairly repeatable fabrication process. Further, these microneedle tips were sharpened by a molding principle, with a simple anisotropic etch of silicon wafer. Also, an enhanced elegant process was explored to incorporate the mounting of the microneedle onto a platform without using any additional material, such that the bore of the microneedle is continuous with the bore of the platform in order to facilitate microfluidic delivery through the hollow needles. The high aspect ratio microneedles undergo failure at the critical load of around 4 N, while the insertion force for such a needle into agar gel, which is a fairly good equivalent of the human skin due to its inherent visco-elastic properties, is 7 mN, which translates into a safety factor (ratio of critical loading force to the maximum applied force) of greater than 500 thus, making it adequately strong for skin penetration.

2-Scale topography dry electrode for biopotential measurements.

The design and fabrication of a novel 2-scale topography dry electrode using macro and micro needles is presented. The macro needles enable biopotential measurements on hairy skin, the function of the micro needles is to decrease the electrode impedance even further by penetrating the outer skin layer. Also, a fast and reliable impedance characterization protocol is described. Based on this impedance measurement protocol, a comparison study is made between our dry electrode, 3 other commercial dry electrodes and a standard wet gel electrode. Promising results are already obtained with our electrodes which do not have skin piercing micro needles. For the proposed electrodes, three different conductive coatings (Ag/AgCl/Au) are compared. AgCl is found to be slightly better than Ag as coating material, while our Au coated electrodes have the highest impedance.

Two-dimensional multi-channel neural probes with electronic depth control.

This paper presents multi-electrode arrays for in vivo neural recording applications incorporating the principle of electronic depth control (EDC), i.e., the electronic selection of recording sites along slender probe shafts independently for multiple channels. Two-dimensional (2D) arrays were realized using a commercial 0.5- µm complementary-metal-oxide-semiconductor (CMOS) process for the EDC circuits combined with post-CMOS micromachining to pattern the comb-like probes and the corresponding electrode metallization. A dedicated CMOS integrated front-end circuit was developed for pre-amplification and multiplexing of the neural signals recorded using these probes.

Design and fabrication of a low cost implantable bladder pressure monitor.

In the frame of the Flemish Community funded project Bioflex we developed and fabricated an implant for short term (< 7 days) bladder pressure monitoring, and diagnosis of incontinence. This implant is soft and flexible to prevent damaging the bladder's inner wall. It contains a standard flexible electronic circuit connected to a battery, which are embedded in surface treated silicone to enhance the biocompatibility and prevent salt deposition. This article describes the fabrication of the pill and the results of preliminary cytotoxicity tests. The electronic design and its tests, implantation and the result of the in-vivo experimentation will be presented in other articles.

Wireless powering for a self-propelled and steerable endoscopic capsule for stomach inspection.

This paper describes the integration of an active locomotion module in a wirelessly powered endoscopic capsule. The device is a submersible capsule optimized to operate in a fluid environment in a liquid-distended stomach. A 3D inductive link is used to supply up to 400mW to the embedded electronics and a set of 4 radio-controlled motor propellers. The design takes advantage of a ferrite-core in the receiving coil-set. This approach significantly improves the coupling with the external field source with respect to earlier work by the group. It doubles the power that can be received with a coreless coil-set under identical external conditions. The upper limit of the received power was achieved complying with the strict regulations for safe exposure of biological tissue to variable magnetic fields. The wireless transferred power was proven to be sufficient to achieve the speed of 7cm/s in any directions. An optimized locomotion strategy was defined which limits the power consumption by running only 2 motors at a time. A user interface and a joystick controller allow to fully drive the capsule in an intuitive manner. The device functionalities were successfully tested in a dry and a wet environment in a laboratory set-up.

A 3D slim-base probe array for in vivo recorded neuron activity.

This paper introduces the first experimental results of a new implantable slim-base three-dimensional (3D) probe array for cerebral applications. The probes are assembled perpendicularly into the slim-base readout platform where electrical and mechanical connections are achieved simultaneously. A new type of micromachined interconnect has been developed to establish electrical connection using extreme planarization techniques. Due to the modular approach of the platform, probe arrays of different dimensions and functionality can be assembled. The platform is only several hundred microns thick which is highly relevant for chronic experiments in which the probe array should be able to float on top of the brain. Preliminary tests were carried out with the implantation of a probe array into the auditory cortex of a rat.

Biomedical stretchable sytems using MID based stretchable electronics technology.

In order to fit human body, flexibility, or even better stretchability is requested for biomedical systems like implants or smart clothes. A stretchable electronic technology has been developed. This can provide highly stretchable interconnections fully compatible with PCB technologies. In order to prove the feasibility of complex biomedical systems like inner body implants or wearable systems, a variety of stretchable systems has been designed from sensor to power source systems.

The influence of micro-motion on the tissue differentiation around immediately loaded cylindrical turned titanium implants.

OBJECTIVE: The aim of this study was to evaluate the effect of various degrees of implant displacement on the tissue differentiation around immediately loaded cylindrical turned titanium implants. DESIGN: The experiments were conducted in repeated sampling bone chambers placed in the tibia of 10 rabbits. Tissues could grow into the bone chambers via perforations. Due to its double structure, tissues inside the chamber could be harvested leaving the chamber intact. This allowed several experiments within the same animal. The chambers contained a cylindrical turned titanium implant that was loaded in a well-controlled manner. In each of the 10 chambers, four experiments were conducted with the following test conditions: immediate implant loading by inducing 0 (control), 30, 60 and 90 microm implant displacement, 800 cycles per day at a frequency of 1 Hz, twice a week during a period of 6 weeks. Histological and histomorphometrical analyses were performed on methylmethacrylate histological sections. An ANOVA was conducted on the dataset. RESULTS: The total tissue volume was significantly lowest in the unloaded control condition. The bone volume fraction on the other hand, was significantly larger in the unloaded and 90 microm implant displacement, compared to the 30 microm implant displacement. Bone density increased with increasing micro-motion with significantly higher values for the 60 microm- and 90 microm-test conditions compared to the unloaded situation. The chance to have bone-to-implant contact decreased in case of micro-motion at the tissues-implant interface. CONCLUSION: The magnitude of implant displacement had a statistically significant effect on the tissue differentiation around immediately loaded cylindrical turned titanium implants. Implant micro-motion had a detrimental effect on the bone-to-implant contact in an immediate loading regimen.

Intracorporeal Videoprobe (IVP).

The objective of the work in the EC project IVP is the development and evaluation of two prototypes of video systems:- a small wired videoprobe with a CMOS image sensorand- an autonomous video-capsule with a telemetric link for image data transfer to an external PC-based system.

Recent progress on transcutaneous energy transfer for total artificial heart systems.

For many years, transcutaneous energy transfer (TET) systems have been developed for energizing total artificial heart systems. Although such a basic system can be developed without too much design effort, optimization toward high power transfer efficiency forces the introduction of novel system topologies and design strategies. In addition, for medical applications, the thermal impact of a TET system on the biological tissue should be taken into account, resulting in limitations on usable coil geometries. This article presents a TET system that has been developed for a power transfer of 25 W over a distance of 1 cm with minimal dimensions of 1 x 6 x 4 cm for the external driver and 5 x 3 x 1 cm for the internal electronics. The coil geometries have a thickness of 2 mm and a diameter of 6 cm. An overall system efficiency of 80% was achieved for an internal load of 25 W.

Pre-load on oral implants after screw tightening fixed full prostheses: an in vivo study.

The fit of implant supported fixed prostheses is said to be of clinical concern because of the rigid fixation of an oral implant in its surrounding bone. The influence of the torque sequence of the set screws during fixation of implant supported fixed full prostheses on the final pre-load was investigated in vitro. No significant effect of the torque sequence of the set screws on the final pre-load was observed. The main objective of this study was to quantify and qualify the pre-load in vivo on implants supporting a fixed full prosthesis. This was performed when the prostheses were supported by all five or six implants and was repeated when the prostheses were supported by only four and three implants. A total of 13 patients with a fixed full implant supported prosthesis were selected. The existing abutments were changed for strain gauged abutments. After tightening the set screws with a torque of 10 N cm, the pre-load conditions were registered. The average (SEM) axial forces and bending moments in case of five or six, four and three supporting implants were 323 N (43 N), 346 N (59 N), 307 N (60 N) 21 N cm (3 N cm) and 21 N cm (2 N cm), 23 N cm (5 N cm), respectively. In addition, the pre-load was registered after fixation of a machined gold cylinder, as delivered by the manufacturer, on each of the supporting implants, representing the 'optimal fit' situation. The corresponding average (SEM) axial forces and bending moments in case of five or six, four and three supporting implants were 426 N (36 N), 405 N (40 N), 413 N (46 N) and 8 N cm (1 N cm), 8 N cm (1 N cm), 8 N cm (1 N cm), respectively. The induced axial forces after tightening the prostheses were significantly lower then after tightening the gold cylinder in case of five or six supporting implants (P < 0.02). The induced bending moments after tightening the prostheses were statistically significantly higher (P < 0.0001) then after tightening the gold cylinder in all test conditions (five or six, four or three supporting implants). This study underlines the static load present after screw tightening implant supported fixed full prostheses.

Three-dimensional force measurements on oral implants: a methodological study.

This paper describes a methodology that allows in vitro and in vivo quantification and qualification of forces on oral implants. Strain gauges are adapted to the outer surface of 5.5 and 7 mm standard abutments (Brånemark System, Nobel Biocare, Sweden). The readings of the strain gauges are transformed into a numerical representation of the normal force and the bending moment around the X- and Y-axis. The hardware and the software of the 3D measuring device based on the strain gauge technology is explained and its accuracy and reliability tested. The accuracy level for axial forces and bending moments is 9.72 N and 2.5 N x cm, respectively, based on the current techniques for strain gauged abutments. As an example, an in vivo force analysis was performed in a patient with a full fixed prosthesis in the mandible. Since axial loads of 450 N and bending moments of 70 N x cm were recorded, it was concluded that the accuracy of the device falls well within the scope of our needs. Nevertheless, more in vivo research is needed before well defined conclusions can be drawn and strategies developed to improve the biomechanics of oral implants.

Magnitude and distribution of occlusal forces on oral implants supporting fixed prostheses: an in vivo study.

Since loading is increasingly believed to be a determining factor in the treatment outcome with oral implants, there is a need to expand the knowledge related to the biomechanics of oral implants. The aim of this study is to gain insight in the distribution and magnitude of occlusal forces on oral implants carrying fixed prostheses. This is done by in vivo quantification and qualification of these forces, which implies that not only the magnitude of the load but also its type (axial force or bending moment) will be registered. A total of 13 patients with an implant supported fixed full prosthesis were selected. Occlusal forces on the supporting implants were quantified and qualified during controlled load application of 50 N on several positions along the occlusal surface of the prostheses and during maximal biting in maximal occlusion by use of strain gauged abutments. The test was conducted when the prostheses were supported by all (5 or 6) implants and was repeated when the prostheses were supported by 4 and by 3 implants only. Despite considerable inter-individual variation, clear differences in implant loading between these test conditions were seen. Loading of the extension parts of the prostheses caused a hinging effect which induced considerable compressive forces on the implants closest to the place of load application and lower compressive or tensile forces on other implants. On average, higher forces were observed with a decreasing number of supporting implants. Bending moments were highest when 3 implants only were used.

Concept, design and fabrication of smart orthopedic implants.

Extensive clinical experience has been built up using orthopaedic implants instrumented with strain gauges connected to a Wheatstone bridge by means of percutaneous leads. This research showed the medical relevance of the monitoring of the deformation of implants as a powerful tool to evaluate nursing and rehabilitation exercises, for tracing dangerous overloads and anticipating implant failure and also to observe the healing process. The IMPACT 3500 project focuses on the instrumentation of femoral implants with on board sensors: regular Benoist-Girard implants have been modified, to contain a 'sensing cell', and thoroughly tested in vitro and in vivo. The implant deformations are measured with resistive strain gauges, and the signal is transferred to a personal computer for processing and display, via a hard wired connection, or via a telemetry system. Two fully implantable wireless designs, called Linkstrain and Sealstrain, are powered from the outside by magnetic induction. As Sealstrain contains the whole telemetric system in its cavity, the highest miniaturization was required; this seriously deteriorates the efficiency of the inductive power link.

Influence of prosthesis material on the loading of implants that support a fixed partial prosthesis: in vivo study.

BACKGROUND: Since loading is increasingly believed to be a determining factor in the treatment outcome with oral implants, there is a need to expand the knowledge related to the biomechanics of oral implants and its influencing factors. PURPOSE: The aim of this study was to investigate the influence of prosthesis material on the distribution and magnitude of load on oral implants carrying a fixed partial prosthesis by in vivo quantification and qualification of this load. METHODS: Eight patients with in total nine three-unit fixed partial prostheses on three implants and three patients with in total four two-unit fixed partial prostheses on two implants were selected. Both metal and acrylic resin prostheses were made. Strain gauged abutments were used to measure the load on the supporting implants during controlled load application of 50 N on several positions along the occlusal surface of the prostheses and during maximal biting in maximal occlusion. Additional tests were conducted when the three-unit prostheses were supported only by two implants, thereby creating an extension pontic. RESULTS: A significantly better distribution of bending moments with the metal prostheses in comparison to the acrylic resin prostheses was observed in the case of the three-unit prostheses on two implants. No other difference in load or load distribution with the different prosthesis materials was noted. CONCLUSION: The clinical significance of the study reveals an increased risk for bending overload of the implants that are closest to the point of load application only in the case of acrylic resin long span prostheses or acrylic resin prostheses with extensions.

In vivo forces on oral implants supporting a mandibular overdenture: the influence of attachment system.

This study was designed to gain insight into the influence of the attachment system on the loading conditions of oral implants supporting a mandibular overdenture on two implants. Five patients were selected and were provided with two implants in the canine area of the mandible (Brånemark System). All patients received a new mandibular overdenture that could be mounted on an ovoid-shaped bar (Dolder, C&M): (a) with and (b) without bilateral extensions and (c) on ball-attachments (Nobel Biocare). Using three strain gauges attached to the outer surface of the 5.5-mm standard abutments, the axial forces and bending moments on both supporting implants could be quantified. Load registrations were made during application of 50 N on seven predetermined positions along the occlusal surface of the prosthesis and during maximal biting in maximal occlusion (clenching). The results revealed no differences in induced axial force for the various anchorage devices, unlike the differences in bending moment. Although there is a tendency for better axial load sharing with bars and better sharing of bending moments with ball attachments, these differences were not significant.

The influence of bone mechanical properties and implant fixation upon bone loading around oral implants.

Finite element models were created to study the stress and strain distribution around a solitary Brånemark implant. The influence of a number of clinically relevant parameters was examined: bone-implant interface (fixed bond versus frictionless free contact), bone elastic properties, unicortical versus bicortical implant fixation and the presence of a lamina dura. Bone loading patterns in the vicinity of the implant seem to be very sensitive to these parameters. Hence they should be integrated correctly in numerical models of in vivo behaviour of oral implants. This necessitates the creation of patient-dependent finite element models.