<p>Recent research has shown that enhanced focal adhesion between cells and extracellular matrix (ECM) and intracellular actin polymerization can accelerate cellular functions like proliferation and differentiation. It is a desirable and necessary technique to modulate cellular functions in the desired lineage in tissue engineering. Previously, we have shown that topographical effects of micropatterns on a cell culture substrate promoted osteogenic differentiation of mesenchymal stem cells (MSCs) without administration of osteogenic growth factors. In this study, bone marrow mesenchymal stem cells from rats were cultured with combined biophysical stimuli, such as surface topography of biomaterials and uniaxial tensile stress and induced osteogenic differentiation. We evaluated MSCs by assessment of alkaline phosphatase (ALP). We demonstrated that a polydimethylsiloxane substrate with microgroove patterns (2 μm of ridge thickness, 1 μm of depth and 1 μm of groove distance) could suppress osteogenic differentiation compared to a flat substrate. Changes of cell adhesion and shape were observed at microgroove substrates by immunofluorescence staining of focal adhesion and actin filaments. We showed that to apply the stretching force promoted differentiation at microgroove substrates but inhibited differentiation at the flat surface. Through this study, a more efficient method to control cellular fate is expected to be established for tissue regeneration <i>in vitro</i>.</p> <div style="text-align:center;"> </div>

Keywords

Mesenchymal stem cells, Osteoblast differentiation, Uniaxial tensile stress, Surface topography, Micropattern, Alkaline phosphatase, Focal adhesion

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[Advance Publication] (Proper information for citation will be announced after formal publication)

Recently, many groups in the field of tissue engineering have attempted to utilize decellularized matrices for tissue regeneration. The decellularized matrices are known as a suitable tissue-engineered scaffold that retains the original structure of extracellular matrix (ECM) in the native tissue as well as its complex vasculature. While chemical reagents such as sodium dodecyl sulfate (SDS) are generally selected to fabricate the decellularized matrices due to its ease of use, high hydrostatic pressure (HHP) has become a powerful alternative to induce the decellularization without using any chemical reagents which have a possibility to provoke inflammatory response by the residual chemicals after <i>in vivo</i> transplantation. Although the HHP has been regarded as a promising tool to decellularize the native tissue, its fabrication mechanism remains still unknown. The aim of this study was to investigate the fabrication mechanism using HHP of 980 MPa to decellularize uterine tissues harvested from Sprague Dawley rats. As a result of histochemical analysis, we first reported that actin filaments in the uterine tissue were depolymerized by applying HHP. Our present findings will lead to the optimization of fabrication method using hydrostatic pressure to have an optimal decellularize matrix with complete micro- and macro-structures of the native tissue for tissue regeneration.

Keywords

Decellularization, Scaffold, Uterus, Hydrostatic pressure, Uterine regeneration

Paper information

Jeonghyun KIM, Shu TAKEDA, Narintadeach CHAROENSOMBUT, Kinyoshi KAWABATA, Yugo KISHIMOTO, Tsuyoshi KIMURA, Akio KISHIDA, Takashi USHIDA, Katsuko S FURUKAWA, “Fabrication of uterine decellularized matrix using high hydrostatic pressure through depolymerization of actin filaments”, Journal of Biomechanical Science and Engineering, Vol.14, No.3 (2019), p.19-00097. doi:10.1299/jbse.19-00097. Final Version Released on November 4, 2019, Advance Publication Released on September 25, 2019.

Adherent cells generate traction forces, which are generated by and transmitted along the actin cytoskeleton to the underlying external substrate via focal adhesions. These cell generated forces are fundamental for maintaining cell shape and driving cell migration as well as triggering signaling pathways to promote processes such as differentiation and proliferation. Here we investigate how mechanical stretch affects traction forces. Following moderate mechanical stretching and release, traction forces are increased and then return to basal levels. However, when cells are stretched with relatively large strains, &gt;10%, traction forces fail to return to basal levels and are attenuated. In this study, we investigate whether cells experiencing attenuated traction forces, following a large strain, can still respond to subsequent mechanical stretching. Traction forces were measured in cells following a consecutive 12% increase in mechanical stretch. We show that, even after traction force attenuation occurs, cells are able to respond to consecutive mechanical stretching by increasing traction force. Visualization of stress fibers, with GFP-actin, indicates that the attenuation of traction force is accompanied by a decrease in stress fiber integrity during mechanical stretching. The ability of cells to increase traction force during a second round of stretching, following attenuation, maybe generated by intact stress fibers that escape damage.

Keywords

Traction force, Mechanical stretch, Actin stress fibers, HUVEC

Paper information

Akira TSUKAMOTO, Katie R. RYAN, Yusuke MITSUOKA, Katsuko S. FURUKAWA, Takashi USHIDA, “Cellular traction forces increase during consecutive mechanical stretching following traction force attenuation”, Journal of Biomechanical Science and Engineering, Vol.12, No.3 (2017), p.17-00118. doi:10.1299/jbse.17-00118. Final Version Released on July 21, 2017, Advance Publication Released on March 06, 2017.

To provide shock-wave exposure with high accuracy and widen the application area of shock-wave therapy, a miniaturized shock-wave device for endoscopic surgery was developed. The shock-wave device applies an electrohydraulic mechanism for generating shock waves in light of device miniaturization and suitability for standard clinical practice. The outer diameter of the shock-wave generator is 11 mm, so it can be inserted in the body through a trocar used in endoscopic surgery or a natural orifice in the case of natural-orifice translumenal endoscopic surgery (NOTES). The distance between the shock-wave focal point and the front of the device is 10 mm. The focused shock wave was observed at the focal point by visualization with the schlieren imaging technique. The pressure at the focal point was measured, and the measurement revealed that the device can produce a peak pressure of 2.32±0.81 MPa at 2-kV discharge voltage, 3.69±1.06 MPa at 3 kV, 5.67±2.44 MPa at 4 kV, and 7.27±2.33 MPa at 5 kV.

Keywords

Underwater Shock Waves, Minimally Invasive Surgery, Forceps-Compatible Shock-Wave Device, Endoscopic Surgery, Electrohydraulic Shock-Wave Generator

Paper information

Keiichi NAKAGAWA, Akira TSUKAMOTO, Tatsuhiko ARAFUNE, Hongen LIAO, Etsuko KOBAYASHI, Takashi USHIDA and Ichiro SAKUMA, “A Miniaturized Shock-Wave Device for Mounting on Forceps”, Journal of Biomechanical Science and Engineering, Vol. 8, No. 1 (2013), pp. 17-26. doi:10.1299/jbse.8.17

Cells under cyclic stretch sense their environments and induce responses such as actin stress fiber (SF) reorientation and morphological changes. These physiological responses are thought to occur when cells sense incompatibility between SF orientation and stretching direction. This hypothesis requires existence of SFs. However, such existence of SFs in cells under cyclic stretch remains unclear since few studies attempted to track the existence of SFs throughout cyclic stretch. In order to track the existence of SFs throughout cyclic stretch, high time resolution time-lapse imaging was improved in two points. First, SFs were clearly imaged with coexpression of DsRed-zyxin and GFP-actin. Second, time resolution was improved so that fluorescence images were obtained every 28 sec. With the improved high time resolution time-lapse imaging, it was revealed, for the first time, that SFs could exist continuously throughout cyclic stretch. Moreover, physiological responses including morphological change as well as SF reorientation occurred during the time when SFs formed incompatibility between SF orientation and stretching direction. These results demonstrated that SFs continuously existed in cells under cyclic stretch and in turn suggested that continuous presence of incompatibility between orientation of long-lasting SFs and the stretching direction might be important for mechanosensing which induces physiological responses.

Keywords

Cyclic Stretch, Stress Fiber, Orientation Dynamics, Morphological Change, Time-Lapse Imaging, HUVEC

Paper information

Yusuke MITSUOKA, Akira TSUKAMOTO, Shunsuke IWAYOSHI, Katsuko S. FURUKAWA and Takashi USHIDA, “High Time Resolution Time-Lapse Imaging Reveals Continuous Existence and Rotation of Stress Fibers under Cyclic Stretch in HUVEC”, Journal of Biomechanical Science and Engineering, Vol. 7, No. 2 (2012), pp.188-198 . doi:10.1299/jbse.7.188

Three-dimensional porous scaffolds play an important role for tissue engineering. The recent developments of porous scaffolds and their preparation methods, especially those developed by our group, are summarized in this review. A method for the preparation of biodegradable porous scaffolds has been developed by using pre-prepared ice particulates as porogen material. A kind of composite biodegradable porous scaffolds has been developed by forming collagen microsponges in the pores or interstices of a synthetic polymer sponge or mesh. A composite sponge of synthetic polymer, collagen and hydroxyapatite has been developed for hard tissue engineering. Bovine articular cartilage-like tissue has been engineered by culturing chondrocytes in the PLGA-collagen scaffolds.

Keywords

Biodegradable Polymer, Tissue Engineering, Composite Sponge, Composite Mesh, Articular Cartilage

Paper information

Tetsuya TATEISHI, Guoping CHEN and Takashi USHIDA, “Polyfunctional Scaffolds for Tissue Engineering”, Journal of Biomechanical Science and Engineering, Vol. 1, No. 1 (2006), pp.8-15 . doi:10.1299/jbse.1.8

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Keywords

Paper information

Takashi USHIDA and Taiji ADACHI, “Preface”, Journal of Biomechanical Science and Engineering, Vol. 1, No. 1 (2006), pp.1-1 . doi:10.1299/jbse.1.1