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Low-intensity pulsed ultrasound activates the phosphatidylinositol 3 kinase/Akt pathway and stimulates the growth of chondrocytes in three-dimensional cultures: a basic science study

Ryohei Takeuchi1*, Akihide Ryo23, Noriko Komitsu1, Yuko Mikuni-Takagaki4, Atsuko Fukui1, Yuta Takagi5, Toshihiko Shiraishi5, Shin Morishita5, Yoshiyuki Yamazaki1, Ken Kumagai1, Ichiro Aoki3 and Tomoyuki Saito1

Author Affiliations

1 Department of Orthopaedic Surgery, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama City, Kanagawa 236-0004, Japan

2 First Research Group, AIDS Research Center, National Institute of Infectious Diseases, 4-7-1 Gagkuen, Musashimurayama, Tokyo 208-0011, Japan

3 Department of Pathology, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama City, Kanagawa 236-0004, Japan

4 Department of Functional Biology, Kanagawa Dental College, 82 Inaokachyo, Yokosuka City, Kanagawa 238-8580, Japan

5 Department of Environment and Information Sciences, Yokohama National University Graduate School, 79-5 Tokiwadai, Hodogaya-ku, Yokohama City, Kanagawa 240-851, Japan

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Arthritis Research & Therapy 2008, 10:R77  doi:10.1186/ar2451

Published: 11 July 2008



The effect of low-intensity pulsed ultrasound (LIPUS) on cell growth was examined in three-dimensional-cultured chondrocytes with a collagen sponge. To elucidate the mechanisms underlying the mechanical activation of chondrocytes, intracellular signaling pathways through the Ras/mitogen-activated protein kinase (MAPK) and the integrin/phosphatidylinositol 3 kinase (PI3K)/Akt pathways as well as proteins involved in proliferation of chondrocytes were examined in LIPUS-treated chondrocytes.


Articular cartilage tissue was obtained from the metatarso-phalangeal joints of freshly sacrificed pigs. Isolated chondrocytes mixed with collagen gel and culture medium composites were added to type-I collagen honeycomb sponges. Experimental cells were cultured with daily 20-minute exposures to LIPUS. The chondrocytes proliferated and a collagenous matrix was formed on the surface of the sponge. Cell counting, histological examinations, immunohistochemical analyses and western blotting analysis were performed.


The rate of chondrocyte proliferation was slightly but significantly higher in the LIPUS group in comparison with the control group during the 2-week culture period. Western blot analysis showed intense staining of type-IX collagen, cyclin B1 and cyclin D1, phosphorylated focal adhesion kinase, and phosphorylated Akt in the LIPUS group in comparison with the control group. No differences were detected, however, in the MAPK, phosphorylated MAPK and type-II collagen levels.


LIPUS promoted the proliferation of cultured chondrocytes and the production of type-IX collagen in a three-dimensional culture using a collagen sponge. In addition, the anabolic LIPUS signal transduction to the nucleus via the integrin/phosphatidylinositol 3-OH kinase/Akt pathway rather than the integrin/MAPK pathway was generally associated with cell proliferation.