Mechanobiology Mechanobiology and its Clinical Application and its - PowerPoint PPT Presentation

Mechanobiology Mechanobiology and its Clinical Application and its Clinical Application Keiji Keiji Naruse Naruse, M.D. & Ph.D. , M.D. & Ph.D. Professor and Chair Professor and Chair Cardiovascular Physiology, Okayama Cardiovascular Physiology, Okayama Univ Univ Grad Grad Sch Sch Med Med CSO, STREX Inc. CSO, STREX Inc. Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

OVERVIEW OVERVIEW MECHANOBIOLOGY MECHANOBIOLOGY MECHANOTRANSDUCTION MECHANOTRANSDUCTION REPRODUCTIVE MEDICINE REPRODUCTIVE MEDICINE Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

We are continually exposed to mechanical stimuli We are continually exposed to mechanical stimuli TEETH TEETH HEARING HEARING TOUCH TOUCH periodontal ligament cells Bone CARDIOVASCULAR CARDIOVASCULAR MECHANOBIOLOGY MECHANOBIOLOGY JOINTS JOINTS REPRODUCTION REPRODUCTION 1xG 1xG Modified from ” ICURUS ” by HENRY MATISSE Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Hemodynamic forces Hemodynamic forces Endothelial cells Endothelial cells shear stress shear stress stretch stretch pressure pressure NO, ET1 etc. NO, ET1 etc. STRETCH STRETCH- -ACTIVATED ACTIVATED Gene expression Gene expression ION CHANNEL ION CHANNEL Morphological changes Morphological changes Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Mechanobiology Mechanobiology Biomedical Biomedical MEMS MEMS Biomedical Engineering Biomedical Engineering SOFT LITHOGRAPHY SOFT LITHOGRAPHY • MEMS( MEMS(micro electro mechanical systems) ) Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Soft Lithography Soft Lithography Mechanotransduction • PDMS PDMS ( (polydimethylsiloxane polydimethylsiloxane) ) • nm nm- -mm mm photolithography photolithography • Merits Merits milling milling Light, EB Light, EB – Transparent Transparent photomask photomask – Stable Stable – Flexible Flexible – High throughput High throughput regist regist – Inexpensive Inexpensive Silicon wafer Silicon wafer PDMS PDMS pore pore cure cure lift off lift off Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Chamber Stretch Chamber Mechanotransduction Stretch chamber Stretch chamber PDMS PDMS mould mould Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch System Stretch System Mechanotransduction Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch- -induced Morphological change induced Morphological change 1 Hz 20 % 1 Hz 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch- Stretch -induced morphological change induced morphological change 20 15 0 min 0 min 10 % 5 0 0 30 60 90 120 150 180 20 15 30 min 30 min 10 % 5 0 0 30 60 90 120 150 180 20 15 10 60 min 60 min % 5 0 0 30 60 90 120 150 180 20 15 10 120 min 120 min % 5 0 0 30 60 90 120 150 180 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch- Stretch -induced morphological change induced morphological change Stretch Stretch- -activated channel dependent activated channel dependent Gd 3+ 3+ Gd 2+ free Gd 3+ 3+ Ca 2+ Gd Ca free 0 min 0 min 20 20 15 15 10 10 % % 5 5 0 0 0 30 60 90 120 150 180 0 30 60 90 120 150 180 20 20 30 min 30 min 15 15 10 10 % % 5 5 2+ free Ca 2+ Ca free 0 0 0 30 60 90 120 150 180 0 30 60 90 120 150 180 20 20 15 15 10 10 60 min 60 min % % 5 5 0 0 0 30 60 90 120 150 180 0 30 60 90 120 150 180 20 20 15 15 10 10 % % 5 5 120 min 120 min 0 0 0 30 60 90 120 150 180 0 30 60 90 120 150 180 1 Hz, 20 % 60 min 1 Hz, 20 % 60 min 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch- -induced Ca response induced Ca response SA channel SA channel Gd blocks SA channel Ca 2+ 2+ influx Ca influx Fura2 Fura2 Fluorsecence Fluorsecence Microscopy Microscopy Plasma membrane Plasma membrane 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch Stretch- -induced Ca response induced Ca response 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch- Stretch -induced Ca response induced Ca response 1.6 1.4 1.2 Ratio (340/380) 1.0 0.8 0.6 (10%) (15%) (20%) (25%) (5%) 0.4 200 400 600 800 0 Stretch for 3 sec Time (sec) Br J Pharmacol, 2002 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

Stretch- Stretch -induced Ca response induced Ca response Stretch- Stretch -activated channel dependent activated channel dependent 1.4 1.2 Stretch for 3 sec Ratio (340/380) 1.0 0.8 (10%) (15%) (20%) (25%) (10%) (15%) (20%) (25%) (5%) (5%) 0.6 Gd 3+ 0.4 0.4 0 0 100 200 300 400 500 600 Time (sec) Br J Pharmacol, 2002 1 Hz, 20 % Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel • MscL MscL, , MscS MscS (Non (Non-selective) selective) • Mid Mid- -1 ( 1 (Non Non- -selective) selective) (K + selective) • TREK/TRAAK TREK/TRAAK ( selective) SAKCA (K + selective) • SAKCA (K selective) • TRPs ( TRPs (cation cation selective) selective) Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine

SA channel SA channel Mid1 Mid1 Molecular Identification of a Eukaryotic, Stretch Molecular Identification of a Eukaryotic, Stretch- - Channel ： Mid1 Activated Nonselective Activated Nonselective Cation Cation Channel Mid1 Science, 285:882-885 1999 Science, 285:882 885 1999 Cardiovascular Physiology Okayama University Graduate School of Medicine Cardiovascular Physiology Okayama University Graduate School of Medicine