Low-intensity pulsed ultrasound (LIPUS) and cell-to-cell communication in bone marrow stromal cells

Sena, Kotaro; Angle, Siddhesh R.; Kanaji, Arihiko; Aher, Chetan V.; Karwo, David G.; Sumner, Dale R.; Virdi, Amarjit S.

doi:10.1016/j.ultras.2011.01.007

Cited by 40 publications

(22 citation statements)

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“…38,39 Many studies examining the effects of LIPUS on cultured cells used similar experimental parameters, such as stimulus power, duration, and ultrasound frequency. [40][41][42][43][44] The focus of this study was to determine whether LIPUS stimulates the expression of HSP leading to osteoblast activation and whether osteoblast stimulation could activate osteogenic signal transduction, independent of the canonical BMP signaling pathway. The experimental protocol used in the present study was based on the precedent manuals, although in this study, LIPUS exposure was concentrated into a one-time treatment at 30 mW/cm 2 for 15 min.…”

Section: Discussionmentioning

confidence: 99%

“…Although different exposures such as 45 and 60 mW/cm 2 were examined rather than 30 mW/cm 2 , those stronger exposures did not accelerate the differentiation or proliferation of the cultured cells, and the design used in the current study just followed similar in vitro studies. 40,42,45 Heat alters the induction of HSP27, HSP70, and HSP90 in osteoblasts. 13 HSP70 expression in cultured human bone marrow cells was increased by the application of mild heat (39°C).…”

Section: Discussionmentioning

confidence: 99%

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Low-Intensity Pulsed Ultrasound Stimulation Enhances Heat-Shock Protein 90 and Mineralized Nodule Formation in Mouse Calvaria-Derived Osteoblasts

Miyasaka

Nakata

Jia

et al. 2015

Tissue Engineering Part A

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Low-intensity pulsed ultrasound (LIPUS) has demonstrated its positive effects on osteogenic differentiation of mesenchymal stem cells and the proliferation and differentiation of osteoblasts, negative effects on osteoclast growth, and promotion of angiogenesis, leading to improvement of the tissue perfusion. Heat-shock proteins (HSPs) are initially identified as molecules encouraged and expressed by heat stress or chemical stress to cells and involved in the balance between differentiation and apoptosis of osteoblasts. However, it remains unclear if the effect of LIPUS on osteoblast differentiation could involve HSP expression and contribution. In this study, mouse calvarial osteoblasts were exposed to LIPUS at a frequency of 3.0 MHz by 30 mW/cm 2 for 15 min or to 42°C heat shock for 20 min at day 3 of cell culture and examined for osteogenesis with pursuing induction of HSP27, HSP70, and HSP90. LIPUS as well as heat shock initially upregulated HSP90 and phosphorylation of Smad1 and Smad5, encouraging cell viability and proliferation at 24 h, enhancing mineralized nodule formation stronger by LIPUS after 10 days. However, HSP27, associated with BMP2-stimulated p38 mitogen-activated protein kinase during osteoblast differentiation, was downregulated by both stimulations at this early time point. Notably, these two stimuli maintained Smad1 phosphorylation with mineralized nodule formation even under BMP2 signal blockage. Therefore, LIPUS might be a novel inducer of osteoblastic differentiation through a noncanonical signal pathway. In conclusion, LIPUS stimulation enhanced cell viability and proliferation as early as 24 h after treatment, and HSP90 was upregulated, leading to dense mineralization in the osteoblast cell culture after 10 days.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Low-Intensity Pulsed Ultrasound Stimulation Enhances Heat-Shock Protein 90 and Mineralized Nodule Formation in Mouse Calvaria-Derived Osteoblasts

Miyasaka

Nakata

Jia

et al. 2015

Tissue Engineering Part A

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“…Taken together, these data suggest that SDF-1/CXCR4 signaling plays a very important role in fracture healing. Disrupting the SDF/CXCR4 pathway during LIPUS treatment could partially reduce, but not fully abolish LIPUS-induced fracture healing, indicating that other factors or signaling pathways might also be involved in this accelerated fracture repair process, such as increased blood circulation [88], upregulated osteogenic genes [89], and gap junctional cell-to-cell intercellular communication in rat MSCs [69].…”

Section: Discussionmentioning

confidence: 99%

Low Intensity Pulsed Ultrasound Enhanced Mesenchymal Stem Cell Recruitment through Stromal Derived Factor-1 Signaling in Fracture Healing

Wei

Leung

et al. 2014

PLoS ONE

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Low intensity pulsed ultrasound (LIPUS) has been proven effective in promoting fracture healing but the underlying mechanisms are not fully depicted. We examined the effect of LIPUS on the recruitment of mesenchymal stem cells (MSCs) and the pivotal role of stromal cell-derived factor-1/C-X-C chemokine receptor type 4 (SDF-1/CXCR4) pathway in response to LIPUS stimulation, which are essential factors in bone fracture healing. For in vitro study, isolated rat MSCs were divided into control or LIPUS group. LIPUS treatment was given 20 minutes/day at 37°C for 3 days. Control group received sham LIPUS treatment. After treatment, intracellular CXCR4 mRNA, SDF-1 mRNA and secreted SDF-1 protein levels were quantified, and MSCs migration was evaluated with or without blocking SDF-1/CXCR4 pathway by AMD3100. For in vivo study, fractured 8-week-old young rats received intracardiac administration of MSCs were assigned to LIPUS treatment, LIPUS+AMD3100 treatment or vehicle control group. The migration of transplanted MSC to the fracture site was investigated by ex vivo fluorescent imaging. SDF-1 protein levels at fracture site and in serum were examined. Fracture healing parameters, including callus morphology, micro-architecture of the callus and biomechanical properties of the healing bone were investigated. The in vitro results showed that LIPUS upregulated SDF-1 and CXCR4 expressions in MSCs, and elevated SDF-1 protein level in the conditioned medium. MSCs migration was promoted by LIPUS and partially inhibited by AMD3100. In vivo study demonstrated that LIPUS promoted MSCs migration to the fracture site, which was associated with an increase of local and serum SDF-1 level, the changes in callus formation, and the improvement of callus microarchitecture and mechanical properties; whereas the blockade of SDF-1/CXCR4 signaling attenuated the LIPUS effects on the fractured bones. These results suggested SDF-1 mediated MSCs migration might be one of the crucial mechanisms through which LIPUS exerted influence on fracture healing.

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“…After trypsinization with 0.25% trypsin-0.91 mM EDTA solution, the cells (5 × 10 5 cells) were seeded on a 35-mm plastic culture dish (ASAHI GLASS Co., Ltd., Tokyo, Japan) with 2 mL of culture medium and cultured at 37 °C for 24 h. It has been demonstrated that LIPUS is applied to cells from the upper [26,38] or lower side [25,27–29,31,33,35–37,39] of the culture dish. As demonstrated in Figure 8, LIPUS was transmitted through the bottom of the culture dish with standard US coupling gel (Teijin Pharma, Ltd., Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…We recently demonstrated that the osteoblast activity in goldfish scales responds sensitively to LIPUS and may be important in promoting bone formation [31]. LIPUS has been proven effective in bone marrow stromal cells, which are thought to be one of the cell types involved in fracture healing [32,33]. In addition, LIPUS is reported to stimulate the growth and synthesis of matrix proteins in chondrocytes [34,35].…”

Section: Introductionmentioning

confidence: 99%

Genes Responsive to Low-Intensity Pulsed Ultrasound in MC3T3-E1 Preosteoblast Cells

Tabuchi

Sugahara

Ikegame

et al. 2013

IJMS

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Although low-intensity pulsed ultrasound (LIPUS) has been shown to enhance bone fracture healing, the underlying mechanism of LIPUS remains to be fully elucidated. Here, to better understand the molecular mechanism underlying cellular responses to LIPUS, we investigated gene expression profiles in mouse MC3T3-E1 preosteoblast cells exposed to LIPUS using high-density oligonucleotide microarrays and computational gene expression analysis tools. Although treatment of the cells with a single 20-min LIPUS (1.5 MHz, 30 mW/cm2) did not affect the cell growth or alkaline phosphatase activity, the treatment significantly increased the mRNA level of Bglap. Microarray analysis demonstrated that 38 genes were upregulated and 37 genes were downregulated by 1.5-fold or more in the cells at 24-h post-treatment. Ingenuity pathway analysis demonstrated that the gene network U (up) contained many upregulated genes that were mainly associated with bone morphology in the category of biological functions of skeletal and muscular system development and function. Moreover, the biological function of the gene network D (down), which contained downregulated genes, was associated with gene expression, the cell cycle and connective tissue development and function. These results should help to further clarify the molecular basis of the mechanisms of the LIPUS response in osteoblast cells.

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Low-intensity pulsed ultrasound (LIPUS) and cell-to-cell communication in bone marrow stromal cells

Cited by 40 publications

References 50 publications

Low-Intensity Pulsed Ultrasound Stimulation Enhances Heat-Shock Protein 90 and Mineralized Nodule Formation in Mouse Calvaria-Derived Osteoblasts

Low-Intensity Pulsed Ultrasound Stimulation Enhances Heat-Shock Protein 90 and Mineralized Nodule Formation in Mouse Calvaria-Derived Osteoblasts

Low Intensity Pulsed Ultrasound Enhanced Mesenchymal Stem Cell Recruitment through Stromal Derived Factor-1 Signaling in Fracture Healing

Genes Responsive to Low-Intensity Pulsed Ultrasound in MC3T3-E1 Preosteoblast Cells

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