Analysis of implant stability changes in immediate loading using a laser displacement sensor in vivo and comparison of its sensitivity with that of resonance frequency analysis.

OBJECTIVE: The aim of this study was to analyze the stability changes in immediately loaded implants by using an in vivo quantitative measurement of micromotion under functional dynamic loading and to verify the sensitivity of Resonance Frequency Analysis (RFA) as compared to that of actual micromotion. MATERIALS AND METHODS: The micromotions of immediately loaded implants placed in the tibia of 11 rabbits were monitored using a laser displacement sensor. Functional dynamic loading forces were applied 5 days a week for 6 weeks. The implant stability quotient (ISQ) was monitored using RFA. RESULTS: The micromotion of the almost-loaded implants increased to peak values the day after loading was started and subsequently reached a plateau gradually. The ISQ changes in the loaded implants closely correlated with the alterations of the actual micromotion (r = -0.98, p < .01). Although the ISQ value itself correlated with the measured micromotion at the time of initial fixation (r = 0.73, p < .05), it did not correlate with the micromotion of the implant that acquired integration. No close correlation was observed between the ISQ and the histomorphometrical data. CONCLUSION: The immediately loaded implants showed the lowest stability immediately after the start of loading, which gradually increased thereafter. RFA is considered a useful method for examining stability changes and initial stability; however, it cannot determine the absolute magnitude of the stability after integration.

Effects of implant tilting and the loading direction on the displacement and micromotion of immediately loaded implants: an in vitro experiment and finite element analysis.

PURPOSE: The purpose of this study was to investigate the effects of implant tilting and the loading direction on the displacement and micromotion (relative displacement between the implant and bone) of immediately loaded implants by in vitro experiments and finite element analysis (FEA). METHODS: Six artificial bone blocks were prepared. Six screw-type implants with a length of 10 mm and diameter of 4.3 mm were placed, with 3 positioned axially and 3 tilted. The tilted implants were 30° distally inclined to the axial implants. Vertical and mesiodistal oblique (45° angle) loads of 200 N were applied to the top of the abutment, and the abutment displacement was recorded. Nonlinear finite element models simulating the in vitro experiment were constructed, and the abutment displacement and micromotion were calculated. The data on the abutment displacement from in vitro experiments and FEA were compared, and the validity of the finite element model was evaluated. RESULTS: The abutment displacement was greater under oblique loading than under axial loading and greater for the tilted implants than for the axial implants. The in vitro and FEA results showed satisfactory consistency. The maximum micromotion was 2.8- to 4.1-fold higher under oblique loading than under vertical loading. The maximum micromotion values in the axial and tilted implants were very close under vertical loading. However, in the tilted implant model, the maximum micromotion was 38.7% less than in the axial implant model under oblique loading. The relationship between abutment displacement and micromotion varied according to the loading direction (vertical or oblique) as well as the implant insertion angle (axial or tilted). CONCLUSIONS: Tilted implants may have a lower maximum extent of micromotion than axial implants under mesiodistal oblique loading. The maximum micromotion values were strongly influenced by the loading direction. The maximum micromotion values did not reflect the abutment displacement values.

The effects of bone density and crestal cortical bone thickness on micromotion and peri-implant bone strain distribution in an immediately loaded implant: a nonlinear finite element analysis.

PURPOSE: This study investigated the effects of bone density and crestal cortical bone thickness at the implant-placement site on micromotion (relative displacement between the implant and bone) and the peri-implant bone strain distribution under immediate-loading conditions. METHODS: A three-dimensional finite element model of the posterior mandible with an implant was constructed. Various bone parameters were simulated, including low or high cancellous bone density, low or high crestal cortical bone density, and crestal cortical bone thicknesses ranging from 0.5 to 2.5 mm. Delayed- and immediate-loading conditions were simulated. A buccolingual oblique load of 200 N was applied to the top of the abutment. RESULTS: The maximum extent of micromotion was approximately 100 µm in the low-density cancellous bone models, whereas it was under 30 µm in the high-density cancellous bone models. Crestal cortical bone thickness significantly affected the maximum micromotion in the low-density cancellous bone models. The minimum principal strain in the peri-implant cortical bone was affected by the density of the crestal cortical bone and cancellous bone to the same degree for both delayed and immediate loading. In the low-density cancellous bone models under immediate loading, the minimum principal strain in the peri-implant cortical bone decreased with an increase in crestal cortical bone thickness. CONCLUSIONS: Cancellous bone density may be a critical factor for avoiding excessive micromotion in immediately loaded implants. Crestal cortical bone thickness significantly affected the maximum extent of micromotion and peri-implant bone strain in simulations of low-density cancellous bone under immediate loading.

Theoretical efficacy of preventive measures for pathologic fracture after surgical removal of mandibular lesions based on a three-dimensional finite element analysis.

PURPOSE: Pathologic fracture of the mandible after removal of a lesion historically has been a clinical problem. The present study aimed to evaluate mandibular strength after removal of a lesion and to illustrate the theoretical efficacy of preventive measures against pathologic fracture based on a 3-dimensional finite element (FE) analysis. MATERIALS AND METHODS: A computed tomographic (CT)-based FE model of the mandible of a patient with a dentigerous cyst including a third molar was constructed. Using this model, the decrease of mandibular strength after virtual removal of the lesion was analyzed. The effect of the decrease of occlusal force and reinforcement by a miniplate was analyzed using a simple FE model of the mandible. Based on these analyses, removal of the cyst with the third molar was performed with a decrease of occlusal force and reinforcement by a miniplate. The validity of these procedures was analyzed using a CT-based FE model constructed after surgery. RESULTS: The von Mises stress in a CT-based FE model after virtual removal of the cyst with the third molar was markedly greater than that in the original FE model. In the analysis using a simple FE model, the stress around the fenestrated area was decreased after premolar loading compared with that after molar loading. In addition, miniplate placement around the fenestrated area markedly decreased the stress. Based on these results, the cast crowns of the first and second molars were removed and the fenestrated area of the mandible was reinforced with a 1.5-mm locking miniplate in the actual surgery. The von Mises stress in the fenestrated area was decreased and primarily borne by the miniplate in the analysis of a CT-based FE model constructed after surgery. CONCLUSION: The present study illustrated the theoretical efficacy of plate application for the decrease of stress on the mandible after surgical removal of a cyst including a third molar based on a simulation by FE analysis.

Effect of clenching on biomechanical response of human mandible and temporomandibular joint to traumatic force analyzed by finite element method.

PURPOSE: The purpose of the present study was to analyze the effect of clenching on the biomechanical response of human mandible and temporomandibular joint (TMJ) to traumatic force by the finite element (FE) method. MATERIAL AND METHODS: FE models of the mandible and the TMJ in resting and clenching positions were prepared. Distribution and magnitude of von Mises stress were analyzed by applying force as a point load in the symphyseal, canine, body and angle regions of the mandible. In addition, strain energy density (SED) at the articular disc and in posterior connective tissue of TMJ was analyzed. RESULTS: In the resting position, von Mises stress was mainly concentrated at the condylar neck and in the retromolar region of the mandible. In the clenching position, the stress at the condylar neck decreased in all loadings. The stress in the retromolar region similary decreased in the symphyseal, canine and body loading, respectively; however, higher stress was observed in the retromolar region on the loading side in the angle loading. High SED was generated at the articular disc and in posterior connective tissues of TMJ in the resting position. The SED in these tissues decreased in all loadings in the clenching position. CONCLUSIONS: Clenching generally reduces stress at the condylar neck and in the retromolar region of the mandible, and strain energy at the articular disc and in posterior connective tissue of TMJ by traumatic forces on the mandible; however, clenching induces greater stress in the retromolar region on the loading side by traumatic force to the angle region.

Biomechanical analysis of the strength of the mandible after marginal resection.

PURPOSE: This study investigated the biomechanical behavior of the mandible after marginal resection by tensile test in a human cadaveric mandible and finite element (FE) analysis. MATERIALS AND METHODS: Human cadaveric mandibular models after marginal resection were prepared with residual heights of 5, 10, and 15 mm. The strength in each of these mandibular models was examined by tensile testing. In addition, FE models of the mandible after marginal resection were prepared with residual heights of 5, 7.5, 10, 12.5, and 15 mm. Distribution and magnitude of von Mises stress were analyzed by applying bite forces of 151 N as a point load on the incisal region and 355.2 and 478.1 N on the premolar and molar regions on the nonresected and resected sides, respectively. At the molar region of the resected side, bite forces of 368.5 N and 286.9 N (80% and 60%, respectively, of 478.1 N) were also applied. RESULTS: On tensile testing, all cadaveric mandibular models were broken at the posterior resection corner. The tensile force was significantly larger in the model with a residual height of 15 mm compared with that of those with a 5- or 10-mm residual height. On FE analysis, von Mises stress was concentrated at the resection corner. The region of maximal von Mises stress concentration in FE models was consistent with that showing destruction on tensile testing. The relationship between the residual height and von Mises stress in the resection area was linear in models of the incisal, premolar, and molar loading on the nonresected side and quadratic in models of the premolar and molar loading on the resected side. The maximal von Mises stress in the resection area was highest during molar loading on the resected side under the present loading condition and exceeded the threshold for the development of pathologic fracture in the model with a residual height of around 10 mm or less. However, the maximal von Mises stress decreased in parallel with the reduction of bite force in the molar region of the resected side. CONCLUSIONS: The residual height and bite force are critical factors for the prevention of pathologic fracture of the mandible after marginal resection. Currently, a residual height of more than 10 mm and reduction of bite force are recommended to reduce the risk of fracture.

Direct 3-D morphological measurements of silicone rubber impression using micro-focus X-ray CT.

Three-dimensional computer models of dental arches play a significant role in prosthetic dentistry. The microfocus X-ray CT scanner has the advantage of capturing precise 3D shapes of deep fossa, and we propose a new method of measuring the three-dimensional morphology of a dental impression directly, which will eliminate the conversion process to dental casts. Measurement precision and accuracy were evaluated using a standard gage comprised of steel balls which simulate the dental arch. Measurement accuracy, standard deviation of distance distribution of superimposed models, was determined as +/-0.050 mm in comparison with a CAD model. Impressions and casts of an actual dental arch were scanned by microfocus X-ray CT and three-dimensional models were compared. The impression model had finer morphology, especially around the cervical margins of teeth. Within the limitations of the current study, direct three-dimensional impression modeling was successfully demonstrated using microfocus X-ray CT.

Development of a high-precision image-processing automatic measurement system for MRI visceral fat images acquired using a binomial RF-excitation pulse.

Development of a rapid and accurate method for visceral fat measurement is an important task, given the recent increase in the number of patients with metabolic syndrome. In this study, we optimized the Fast Low Angle Shot (FLASH) sequence using a binominal radiofrequency excitation pulse, in which the acquisition time is short, and measured changes in the amount of visceral fat in subjects after a period of wearing clothes with a fat-reducing effect during walking. We solved the reproducibility problem associated with the number of slices, and developed automatic measurement software for high-precision separation and extraction of abdominal visceral fat images. This software was developed using intensity correction with the coil position, derivation of a threshold by histogram analysis and fat separation by template matching for abdominal images. The cross-sectional area of a single slice varies for every acquisition due to visceral organ movement, but the relative error largely converged for seven slices. The measured amount of abdominal fat tended to be consistent with changes in the body fat and waist circumference of the subjects. The correlation coefficients between automatic extraction using the measurement software and manual extraction were 0.9978 for subcutaneous fat and 0.9972 for visceral fat, showing very strong positive correlations. The consistency rates were 0.9502+/-0.0167 for subcutaneous fat and 0.9395+/-0.0147 for visceral fat, and the shapes of the regions were also extracted very accurately. These results show that the magnetic resonance imaging acquisition method and image processing system developed in this study are beneficial for measurement of abdominal visceral fat. Therefore, this method may have a major role in future diagnosis of metabolic syndrome.

Trend report on international and Japanese standardization activities for bioceramics and tissue engineered medical products.

Since porous and injectable bioceramics have recently been utilized often as scaffolds for bone regenerative medicine, the need for their standardization has increased. One of the standard proposals in ISO/TC150 and JIS has been a draft for characterization of the porous bioceramic scaffolds in both micro- and macro-scopic aspects. ISO/TC150/SC7 (Tissue engineered medical products) has been co-chaired by Professor J E Lemons, Department of Surgery, University of Alabama at Birmingham and Dr R Nakaoka, Division of Medical Devices, National Institute of Health Sciences, Japan. The scope of SC7 has been specified as 'Standardization for the general requirements and performance of tissue engineered medical products with the exclusion of gene therapy, transplantation and transfusion'.

Effect of fracture gap on stability of compression plate fixation: a finite element study.

In compression plating, anatomical reduction and compression across the fracture site are the basic principles necessary to achieve primary bone healing. However, varying amounts of gap at the fracture site frequently occur due to technical pitfalls, such as overbending of the plate and inaccurate reduction, and due to the fracture configuration itself. Little is known as to how fracture gap affects stability of the bone-plate construct. We analyzed the effects of fracture gap size (1 and 4 mm) and bone defect (25%, 50%, 75%, 100%) on the biomechanical stability of the compression plate-bone construct through validated finite element analysis. The stiffnesses of eight different models were compared with the stiffness of an ideally compressed model (0 mm/0%). Stress concentration in form of peak von Mises stress (PVMS) was also evaluated. The decrease in stiffness depended mainly on the depth of bone defect. The decrease in stiffness was similar in models with the same defect and different gap size. Considerably more stress was concentrated around the central hole of the plate in gap models with the depth of bone defects of 75% and 100% than with smaller defects. We concluded that even a thin fracture gap (1 mm) with no contact between the fracture after plating decreases stiffness exponentially; contact at the fracture surfaces of > or =50% was necessary to avoid undue stress concentration in the plate.

Control of proliferation and differentiation of osteoblasts on apatite-coated poly(vinyl alcohol) hydrogel as an artificial articular cartilage material.

One of the key challenges in employing biomaterials is determining how to fix them into the surrounding tissue. To enhance the interaction with surrounding bone, amorphous hydroxyapatite (HA) was coated onto the surface of the bio-inert poly(vinyl alcohol) hydrogel (PVA-H), as an artificial cartilage material, by a pulsed laser deposition technique. Next we examined the binding effects of the HA thin film (300 nm thick) to the underlying bone using osteoblast proliferation and differentiation. A mouse osteoblast cell line, MC3T3E1, was cultured on the HA-coated and noncoated PVA-H with a water content of 33% or 53% for 3 weeks. Cell proliferation, alkaline phosphatase (ALP) activity, and levels of osteocalcin were evaluated for biocompatibility and differentiation. HA coating enhanced the cell proliferation, the ALP activity, and the levels of osteocalcin on both low and high water-content PVA-Hs. The cell growth rates on the PVA-H were lower than on tissue culture dishes even after the HA coating was added; however, osteoblastic differentiation was highly promoted by the HA coating on low water content PVA-H. These results suggested that the HA coating on the PVA-H enhanced the affinity between the bone and the PVA-H as an artificial cartilage material in surface replacement arthroplasty.

Biomechanical analysis of miniplate osteosynthesis for fractures of the atrophic mandible.

PURPOSE: The purpose of this study was to investigate the biomechanical behavior of miniplate osteosynthesis for fracture of the edentulous mandible with various degrees of atrophy by finite element (FE) analysis. MATERIALS AND METHODS: Three-dimensional FE models simulating various atrophic or nonatrophic edentulous mandibles were constructed. The models were divided into 3 groups based on the height: 20 mm, 15 mm, and 10 mm. A model 30-mm high was defined as a nonatrophic mandible. Fracture in the premolar region was simulated. Single or double miniplate osteosynthesis was assumed to fix the fracture. In each case, models of fractures with and without bone contact between bone fragments were prepared. A bite force of 62.8 N was applied in the FE models as a point load on the anterior point. RESULTS: There were no noticeable differences in compressive stress level in the bone around screws among the single miniplate models or double miniplate models with bone contact. Single miniplate models without bone contact showed markedly greater compressive stress than that of models with bone contact. The use of double miniplates showed a great influence on von Mises stress reduction in the miniplates. Without bone contact, greater interfragmentary displacements occurred; however, interfragmentary displacements were within the limit of not causing a malunion of the fractured bone in all models. CONCLUSION: Double miniplate fixation may be a reliable method for treating fracture of the atrophic mandible from a biomechanical viewpoint.

Beneficial storage effects of epigallocatechin-3-o-gallate on the articular cartilage of rabbit osteochondral allografts.

A fresh osteochondral allograft is one of the most effective treatments for cartilage defects of the knee. Despite the clinical success, fresh osteochondral allografts have great limitations in relation to the short storage time that cartilage tissues can be well-preserved. Fresh osteochondral grafts are generally stored in culture medium at 4 degrees C. While the viability of articular cartilage stored in culture medium is significantly diminished within 1 week, appropriate serology testing to minimize the chances for the disease transmission requires a minimum of 2 weeks. (-)-Epigallocatechin-3-O-gallate (EGCG) has differential effects on the proliferation of cancer and normal cells, thus a cytotoxic effect on various cancer cells, but a cytopreservative effect on normal cells. Therefore, a storage solution containing EGCG might extend the storage duration of articular cartilages. Rabbit osteochondral allografts were performed with osteochondral grafts stored at 4 degrees C in culture medium containing EGCG for 2 weeks and then the clinical effects were examined with macroscopic and histological assessment after 4 weeks. The cartilaginous structure of an osteochondral graft stored with EGCG was well-preserved with high cell viability and glycosaminoglycan (GAG) content of the extracellular matrix (ECM). After an osteochondral allograft, the implanted osteochondral grafts stored with EGCG also provided a significantly better retention of the articular cartilage with viability and metabolic activity. These data suggest that EGCG can be an effective storage agent that allows long-term preservation of articular cartilage under cold storage conditions.

Preservation of platelets by adding epigallocatechin-3-o-gallate to platelet concentrates.

The effect of epigallocatechin-3-O-gallate (EGCG), a major component of green tea, on platelet preservation was evaluated. Single donor platelets (N = 10) were collected and preserved by the standard method. EGCG was added to the platelet concentrates before preservation and then the functional and biochemical parameters were monitored throughout the storage period. After 6 days of preservation, the aggregability of the platelets was significantly maintained by addition of 50 and 100 microg/ml of EGCG. Platelet prothrombinase activity was also significantly retained by the addition of EGCG. The accumulation of P-selectin and RANTES in the plasma preserved with EGCG was less than those preserved without EGCG, which indicated that EGCG might inhibit platelet activation. Furthermore, EGCG reduced the increase of LDH in plasma during preservation and inhibited the activation of caspase-3 and cleavage of gelsolin, thereby showing that EGCG could inhibit the apoptosis of platelets. These results suggest that EGCG may play an effective role in preserving platelets by inhibiting the activation and apoptosis of platelets.

Trypsin as a novel potential absorption enhancer for improving the transdermal delivery of macromolecules.

OBJECTIVES: The aim was to assess the effect of trypsin on the transdermal delivery of macromolecules by applying its specific biochemical properties to the stratum corneum of the skin. METHODS: Fluorescein isothiocyanate (FITC)-labelled dextrans (FDs), with molecular weights of 4 to 250 kDa, and FITC-insulin were used as model macromolecules and a model polypeptide, and the in-vitro transdermal permeation experiments, with or without trypsin (0.1-2.5%), were carried out using rat skin and cultured human epidermis. The mechanism for the enhancement of trypsin was also studied using fluorescence and conventional light microscopy. KEY FINDINGS: Trypsin significantly increased the transdermal permeability of all FDs through the rat skin (2.0- to 10.0-fold). It also markedly enhanced the permeation of FD4 through three-dimensional cultured human epidermis (3.1-fold), which was used to evaluate the transport pathways other than the transfollicular route. Furthermore, the permeation flux of FITC-insulin was increased by 10.0-fold with trypsin pretreatment (from 0.02 +/- 0.00 to 0.20 +/- 0.07 microg/cm(2) per h). Mechanistic studies indicated that trypsin affects both the intercellular pathway and the hair follicular route, and may alter stratum corneum protein structures, thereby affecting skin barrier properties. CONCLUSIONS: This study suggests that trypsin could be effective as a biochemical enhancer for the transdermal delivery of macromolecules including peptide and protein drugs.

In vivo structural analysis of articular cartilage using diffusion tensor magnetic resonance imaging.

PURPOSE: The articular cartilage is a small tissue with a matrix structure of three layers between which the orientation of collagen fiber differs. A diffusion-weighted twice-refocused spin-echo echo-planar imaging (SE-EPI) sequence was optimized for the articular cartilage, and the structure of the three layers of human articular cartilage was imaged in vivo from diffusion tensor images. MATERIALS AND METHODS: The subjects imaged were five specimens of swine femur head after removal of the flesh around the knee joint, five specimens of swine articular cartilage with flesh present and the knee cartilage of five adult male volunteers. Based on diffusion-weighted images in six directions, the mean diffusivity (MD) and the fractional anisotropy (FA) values were calculated. RESULTS: Diffusion tensor images of the articular cartilage were obtained by sequence optimization. The MD and FA value of the specimens (each of five examples) under different conditions were estimated. Although the articular cartilage is a small tissue, the matrix structure of each layer in the articular cartilage was obtained by SE-EPI sequence with GRAPPA. The MD and FA values of swine articular cartilage are different between the synovial fluid and saline. In human articular cartilage, the load of the body weight on the knee had an effect on the FA value of the surface layer of the articular cartilage. CONCLUSION: This method can be used to create images of the articular cartilage structure, not only in vitro but also in vivo. Therefore, it is suggested that this method should support the elucidation of the in vivo structure and function of the knee joint and might be applied to clinical practice.

Development of new nerve guide tube for repair of long nerve defects.

A novel nerve guide tube (poly (L-lactic) acid (PLLA)/ polyglycolic acid (PGA)-c-tube) capable of repairing long peripheral nerve injuries in a canine model has been developed. The tube was created by braiding together PLLA and PGA and then coating it with collagen. PLLA was newly added to the formulation to achieve higher sustainability. The tube was compared with a PGA-collagen tube in clinical use since 2002 having the same structure with a collagen coating but composed of PGA alone (PGA-c-tube). When tested for repair of a 40-mm gap in the left peroneal nerve, using PLLA/PGA-c-tube (n = 15), PGA-c-tube (n = 15), and a negative control group where the cut stump was capped using a silicone cap (n = 15), the lumen structure essential for securing the space for nerve regeneration was maintained in PLLA/PGA-c-tube for over 12 months with a higher number of axons both within the tube and at the distal nerve end. Electrophysiological evaluation revealed that the amplitude of compound muscle action potentials and sensory nerve action potentials after nerve regeneration with PLLA/PGA-c-tube were significantly higher. When assessed using magnetic resonance imaging (MRI), the volume of the tibialis anterior (TA) muscle in dogs that had undergone nerve repair using PLLA/PGA-c-tube was approximately 80% that of the positive control at 12 months. Functional analysis conducted by assessing the ankle angle revealed faster recovery in the PLLA/PGA-c-tube group. Better regeneration was achieved using a PLLA/PGA-c-tube that contains the slowly decomposing fiber material, PLLA. This indicates potential for repair of even longer nerve gaps or defects located near joints, and also clinical application.

Analysis of the mandibular movement by simultaneous multisection continuous ultrafast MRI.

PURPOSE: Using simultaneous multiple cross-sectional imaging, we imaged four cross sections, including the mandibular midline and the right and left temporomandibular joints, to observe one movement of mouth opening and closing and analyze the movement of the mandible and temporomandibular joints. MATERIALS AND METHODS: Four cross sections, including the midsagittal section of the mandible, the sagittal sections of the right and left temporomandibular joints and the horizontal section containing the heads of right and left temporomandibular joints, were imaged simultaneously. The imaging was conducted in 10 male and female volunteers. RESULTS: In all volunteers, the relationship of the mandibular movement on the median line with the right and left temporomandibular joints was observed. Images of the volunteers with trismus indicated the condition in which the right and left temporomandibular joints did not move in keeping with each other but moved independently from each other. CONCLUSION: Complex movement of the temporomandibular joint was first evaluated by simultaneous multiple cross-sectional MRI for the movement of mandible and temporomandibular joints.

3D morphological measurements of dental casts with occlusal relationship using microfocus X-ray CT.

In the diagnosis of dental occlusion, it is necessary to quantitatively measure interocclusal contacts and transfer them to a computer model. In this aspect, three-dimensional computer models of upper and lower dental casts play a significant role. In this study, we proposed a new method to measure occlusal interaction by using a microfocus X-ray CT technique. Measurement accuracy was determined as +/-0.03 mm in comparison with a coordinate measuring machine. A superimposition procedure for two sets of three-dimensional dental cast models was also established. Using the same dental cast, the standard deviation between the two sets of models was +/-0.015 mm - which was defined as measurement precision. Between an optical laser scanner and the microfocus X-ray CT system, the standard deviation measured between the two models was +/-0.05 mm. Data were acquired when upper and lower dental casts mounted on the bite impression were scanned, and then occlusal interaction, contacts, and distance distribution between the casts were visualized by a colored map on the cast models. Within the limitations of the current study, it was successfully demonstrated that microfocus Xray CT was well poised for quantitative measurement of occlusal interaction.

Transdermal delivery of insulin using trypsin as a biochemical enhancer.

Trypsin has been extensively used in laboratory settings for in vitro epidermal separation and keratinocyte isolation for over 50 years. The aim of this study was to assess the enhancing effect of trypsin on the transdermal delivery of insulin by applying its specific biochemical properties to react with the stratum corneum (SC) of skin. Bovine insulin was used as a model peptide to investigate in vitro permeation through rat skin and in vivo hypoglycemic effects of bovine insulin with or without the trypsin pretreatment. Trypsin significantly increased the transdermal permeability of bovine insulin in pH 3.0 solution, but no effect was observed in pH 6.0 solution. The permeation flux of bovine insulin from pH 3.0 solution was promoted 5.2-fold with 0.25% trypsin pretreatment when compared with the control. The enhancement of trypsin was dependent on the concentration in the range of 0.5-2.5%. Furthermore, with trypsin pretreatment, the plasma glucose level was reduced to less than 60% of the initial value after 8 h of in vivo permeation of bovine insulin with pH 3.0 solution, but did not return to the initial value during an 8-h experiment. Mechanistic studies with Fourier transform-infrared and attenuated total reflectance analysis and electrical resistance measurements suggest that trypsin alters the SC protein structure from the alpha- to the beta-form and decreases the electrical resistance of skin, thereby decreasing the SC barrier and enhancing the permeation of insulin. We conclude that trypsin would be effective as a biochemical enhancer for the transdermal delivery of peptide and protein drugs such as insulin.