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.
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.
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DOI：
10.1186/ar2451
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发表时间：
2008
影响因子：
4.9
通讯作者：
Saito T
中科院分区：
文献类型：
--
作者：
Takeuchi R;Ryo A;Komitsu N;Mikuni-Takagaki Y;Fukui A;Takagi Y;Shiraishi T;Morishita S;Yamazaki Y;Kumagai K;Aoki I;Saito T
低强度脉冲超声（LIPUS）对胶原海绵三维培养软骨细胞生长的影响。为了阐明软骨细胞机械活化的机制，我们在lipus处理的软骨细胞中检测了Ras/丝裂原活化蛋白激酶（MAPK）和整合素/磷脂酰肌醇3激酶(PI3K)/Akt通路的细胞内信号通路以及参与软骨细胞增殖的蛋白。关节软骨组织取自刚宰杀的猪跖指关节。将分离软骨细胞与胶原凝胶和培养基复合材料混合，加入i型胶原蜂窝海绵中。实验细胞每天暴露于LIPUS 20分钟。软骨细胞增生，海绵表面形成胶原基质。进行细胞计数、组织学检查、免疫组化分析和免疫印迹分析。在2周的培养期间，LIPUS组的软骨细胞增殖率略高于对照组，但明显高于对照组。Western blot分析显示，与对照组相比，LIPUS组的ix型胶原蛋白、细胞周期蛋白B1和细胞周期蛋白D1、局灶黏附激酶磷酸化和Akt磷酸化程度较高。然而，在MAPK、磷酸化MAPK和ii型胶原蛋白水平上没有检测到差异。LIPUS在胶原蛋白海绵三维培养中促进培养软骨细胞的增殖和ix型胶原蛋白的产生。此外，合成代谢LIPUS信号通过整合素/磷脂酰肌醇3-OH激酶/Akt途径而不是整合素/MAPK途径传递到细胞核，通常与细胞增殖有关。
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.
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影响因子：
2.4
作者：
Itoh, H;Aso, Y;Miyata, T
通讯作者：
Miyata, T
影响因子：
2.8
作者：
Parvizi, J;Parpura, V;Bolander, ME
通讯作者：
Bolander, ME
影响因子：
4.8
作者：
Li, Tian-Fang;Chen, Di;O'Keefe, Regis J.
通讯作者：
O'Keefe, Regis J.
影响因子：
--
作者：
Takeuchi, Ryohei;Saito, Tomoyuki;Morishita, Shin
通讯作者：
Morishita, Shin
影响因子：
2.8
作者：
SAH, RLY;KIM, YJ;SANDY, JD
通讯作者：
SANDY, JD