Physical and biological properties of a novel anti-adhesion material made of thermally cross-linked gelatin film: Investigation of the usefulness as anti-adhesion material.

To create more useful, effective and safer anti-adhesion materials, we developed a thermally cross-linked gelatin film. In this study, we examined the physical properties of the film such as the physical strength and the adhesiveness to reveal the handling properties and biological properties, such as the anti-adhesion effect, the influence on cell proliferation, and the cytotoxicity to reveal the anti-adhesion mechanism, especially in comparison with the conventional hyaluronic acid and carboxymethylcellulose film (the conventional film). A tensile test under dry and wet conditions and shearing stress test showed that the gelatin film has significant higher maximum tensile stress and fracture strain than the conventional film. In the study using a rat model of cecum adhesion, the anti-adhesion effect of the gelatin film was significantly superior to that of the conventional film. In the cell proliferation test, the number of fibroblast cells on the gelatin film increased at each time point, while no cell proliferation was observed on the conventional film. Furthermore, in the cytotoxicity test using a colony assay and Live/Dead assay, the extract of the gelatin film had no cytotoxicity, while the extract of the conventional film had cytotoxicity considerably. These results suggest that the gelatin film provides better handling than the conventional film, due to better physical strength and ductility of the film. In addition, the gelatin film has a significantly greater anti-adhesion effect than the conventional film without any cytotoxicity. Therefore, the gelatin film is quite favorable as an anti-adhesion material. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 689-696, 2018.

Multiphoton microscope measurement-based biphasic multiscale analyses of knee joint articular cartilage and chondrocyte by using visco-anisotropic hyperelastic finite element method and smoothed particle hydrodynamics method.

The articular cartilage of a knee joint has a variety of functions including dispersing stress and absorbing shock in the tissue and lubricating the surface region of cartilage. The metabolic activity of chondrocytes under the cyclic mechanical stimulations regenerates the morphology and function of tissues. Hence, the stress evaluation of the chondrocyte is a vital subject to assess the regeneration cycle in the normal walking condition and predict the injury occurrence in the accidents. Further, the threshold determination of stress for the chondrocytes activation is valuable for development of regenerative bioreactor of articular cartilage. In this study, in both macroscale and microscale analyses, the dynamic explicit finite element (FE) method was used for the solid phase and the smoothed particle hydrodynamics (SPH) method was used for the fluid phase. In the homogenization procedure, the representative volume element for the microscale finite element model was derived by using the multiphoton microscope measured 3D structure comprising 3 different layers: surface, middle, and deep layers. The layers had different anisotropic structural and rigidity characteristics because of the collagen fiber orientation. In both macroscale and microscale FE analyses, the visco-anisotropic hyperelastic constitutive law was used. Material properties were identified by experimentally determined stress-strain relationships of 3 layers. With respect to the macroscale and microscale SPH models for non-Newtonian viscous fluid, the previous observation results of interstitial fluid and proteoglycan were used to perform parameter identifications. Biphasic multiscale FE and SPH analyses were conducted under normal walking conditions. Therefore, the hydrostatic and shear stresses occurring in the chondrocytes caused by the compressive load and shear viscous flow were evaluated. These stresses will be used to design an ex-vivo bioreactor to regenerate the damaged articular cartilage, where chondrocytes are seeded in the culture chamber. To know the stress occurred on and in the chondrocytes is vitally important not only to understand the normal metabolic activity of the chondrocyte but also to develop a bioreactor of articular cartilage regeneration as the knee joint disease treatment.

Multi-scale finite element analyses for stress and strain evaluations of braid fibril artificial blood vessel and smooth muscle cell.

In this study, we developed a multi-scale finite element (FE) analysis code to obtain the stress and strain that occurred in the smooth muscle cell (SMC) at micro-scale, which was seeded in the real fabricated braid fibril artificial blood vessel. This FE code can predict the dynamic response of stress under the blood pressure loading. We try to establish a computer-aided engineering (CAE)-driven scaffold design technique for the blood vessel regeneration. Until now, there occurred the great progresses for the endothelial cell activation and intima layer regeneration in the blood vessel regeneration study. However, there remains the difficulty of the SMC activation and media layer regeneration. Therefore, many researchers are now studying to elucidate the fundamental mechanism of SMC activation and media layer regeneration by using the biomechanical technique. As the numerical tool, we used the dynamic-explicit FE code PAM-CRASH, ESI Ltd. For the material models, the nonlinear viscoelastic constitutive law was adapted for the human blood vessel, SMC and the extra-cellular matrix, and the elastic law for the polyglycolic acid (PGA) fiber. Through macro-FE and micro-FE analyses of fabricated braid fibril tubes by using PGA fiber under the combined conditions of the orientation angle and the pitch of fiber, we searched an appropriate structure for the stress stimulation for SMC functionalization. Objectives of this study are indicated as follows: 1. to analyze the stress and strain of the human blood vessel and SMC, and 2. to calculate stress and strain of the real fabricated braid fibril artificial blood vessel and SMC to search an appropriate PGA fiber structure under combined conditions of PGA fiber numbers, 12 and 24, and the helical orientation angles of fiber, 15, 30, 45, 60, and 75 degrees. Finally, we found a braid fibril tube, which has an angle of 15 degree and 12 PGA fibers, as a most appropriate artificial blood vessel for SMC functionalization.

Development of blood extraction system for health monitoring system.

We focus on research to develop a compact human blood sampling device used for the Self Monitoring of Blood Glucose (SMBG). The SMBG comprises: (1) an indentation system using a shape memory alloy (SMA) actuator to force a microneedle through the skin; (2) a micro electrical pumping system to extract blood using a bimorph type piezoelectric microactuator; (3) a biosensor using an enzyme such as glucose oxidase (GOx) to detect and evaluate the amount of glucose in extracted blood. A titanium microneedle the same size as a female mosquito's labium (60 mum outer diameter, 25 mum inner diameter) was produced by the sputter deposition method. The mechanical design of the device was based upon the mosquito's blood sampling mechanism. The blood extraction system worked well. We measured the performance of the principal components: the indentation load for a microneedle of external diameter 100 mum was found to be 0.1 N. The pumping system has an extraction speed of about 2 mul/min for whole blood. This is similar to that achieved by the mosquito.