I–2 Low Intensity Pulsed Ultrasound (LIPUS) Helps to Maintain the Undifferentiated Status of Mesenchymal Stem Cells

Kusuyama, Joji; Seong, Chang Hwan; Bandow, Kenjiro; Kakimoto, Kyoko; Ohnishi, Tomokazu; Matsuguchi, Tetsuya

doi:10.1097/01.bot.0000462953.87235.74

Cited by 4 publications

(6 citation statements)

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“…The phenotypic characterization permitted to identify a functional multipotent hMSC stemness potency maintenance (CD73+/CD90+/CD105+) in the tested groups until 14 days, which decreased at 21 days concurrently with osteoblast differentiation (Kern & Shibata, ; Pittenger et al, ; Ringdén & Le Blanc, ; Rutten et al, ; Saalbach, Haustein, & Anderegg, ; Yoshimura et al, , ). In addition, LIPUS stimulation markedly promoted the expression of stem cell transcriptional factors OCT‐3/4, SOX2, and NANOG related to stemness maintenance, as recently reported by Kusuyama et al (Alvarez et al, ; Kusuyama et al, ). As for the tendency of MSCs to lose their multipotency over time in culture, flow cytometry data recorded at different experimental time points might be consistent with long term culture stress, as reported by Stolzing, Coleman, & Scutt, .…”

Section: Discussionsupporting

confidence: 77%

“…Results of MAPK1 (a), MAPK6 (b) gene expression of Untreated OM ( ), LIPUS MSCGM ( ), and LIPUS OM ( ) cells expressed as relative fold changes (RF) of Untreated MSCGM values (1) (Mean ± SD, n = 3 duplicates). GLM analysis with "group" and "experimental time" as fixed effects showed for: MAPK1-an interaction between "group" and "experimental time," F = 5.92, p < 0.005; MAPK6-an interaction between "group" and "experimental time," F = 4.37, p < 0.005 expression of stem cell transcriptional factors OCT-3/4, SOX2, and NANOG related to stemness maintenance, as recently reported byKusuyama et al (Alvarez et al, 2015;Kusuyama et al, 2015). As for the tendency of MSCs to lose their multipotency over time in culture, flow cytometry data recorded at different experimental time points might be consistent with long term culture stress, as reported byStolzing, Coleman, & Scutt, 2006.…”

supporting

confidence: 70%

“…Recently, it has been showed that low intensity pulsed ultrasound (LIPUS) is able to positively influence the maintenance of MSC stemness (Kusuyama et al, 2015). LIPUS stimulation at 30 mW/cm 2 is an established, widely used and FDA-approved therapeutic treatment for accelerating bone healing in fractures and in delayed or established nonunions (Angle, Sena, Sumner, & Virdi, 2011;Augat, Simon, Liedert, & Claes, 2005;El-Mowafi & Mohsen, 2005).…”

mentioning

confidence: 99%

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Osteogenic commitment and differentiation of human mesenchymal stem cells by low‐intensity pulsed ultrasound stimulation

Costa¹,

Carina²,

Fontana

et al. 2017

Journal Cellular Physiology

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Low-intensity pulsed ultrasound (LIPUS) as an adjuvant therapy in in vitro and in vivo bone engineering has proven to be extremely useful. The present study aimed at investigating the effect of 30 mW/cm LIPUS stimulation on commercially available human mesenchymal stem cells (hMSCs) cultured in basal or osteogenic medium at different experimental time points (7, 14, 21 days). The hypothesis was that LIPUS would improve the osteogenic differentiation of hMSC and guarantying the maintenance of osteogenic committed fraction, as demonstrated by cell vitality and proteomic analysis. LIPUS stimulation (a) regulated the balance between osteoblast commitment and differentiation by specific networks (activations of RhoA/ROCK signaling and upregulation of Ribosome constituent/Protein metabolic process, Glycolysis/Gluconeogenesis, RNA metabolic process/Splicing and Tubulins); (b) allowed the maintenance of a few percentage of osteoblast precursors (21 days CD73+/CD90+: 6%; OCT-3/4+/NANOG+/SOX2+: 10%); (c) induced the activation of osteogenic specific pathways shown by gene expression (early: ALPL, COL1A1, late: RUNX2, BGLAP, MAPK1/6) and related protein release (COL1a1, OPN, OC), in particular in the presence of osteogenic soluble factors able to mimic bone microenvironment. To summarize, LIPUS might be able to improve the osteogenic commitment of hMSCs in vitro, and, at the same time, enhance their osteogenic differentiation.

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Section: Discussionsupporting

confidence: 77%

supporting

confidence: 70%

mentioning

confidence: 99%

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Osteogenic commitment and differentiation of human mesenchymal stem cells by low‐intensity pulsed ultrasound stimulation

Costa¹,

Carina²,

Fontana

et al. 2017

Journal Cellular Physiology

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“…The search for a method that can upregulate and maintain the stemness genes of MSCs is an urgent question. Soluble reagents, cell density, 3D collagen scaffolds, and cell‐specific component media are increasingly considered to be essential for maintaining the stemness of MSCs, and all of these are available in vitro culture settings 5,33–35 . Several studies have shown that low‐intensity pulse US increases MSC proliferation through activation of MAPK/ERK and PI3K/Akt signaling pathways, leading to the upregulation of multiple proteins 31,36 .…”

Section: Discussionmentioning

confidence: 99%

Optimal Treatment Parameters for Ultrasound‐Stimulated Microbubbles in Upregulating Proliferation and Stemness of Bone Marrow Mesenchymal Stem Cells

Chen,

Zhu,

Duan

et al. 2024

J of Ultrasound Medicine

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ObjectivesUltrasound‐targeted microbubble disruption (UTMD) is a widely used technique to improve the differentiation and proliferation capacity of mesenchymal stem cells (MSCs), but the optimal therapeutic parameters for UTMD are unclear. In this study, we aimed to find the appropriate peak negative pressure (PNP), which is a key parameter for enhancing the stemness properties and proliferation of MSCs.MethodsExperiments were performed in UTMD group, ultrasound (US) group under different PNP exposure conditions (0.5, 1.0, and 1.5 MPa), and control group. Apoptosis safety was analyzed by flow cytometry and MSC proliferation was measured at 12, 24, and 36 hours after irradiation by cell counting kit 8. The expression of the stemness genes NANOG, OCT‐4, and SOX‐2 were determined by enzyme‐linked immunosorbent assay (ELISA) or reverse transcription polymerase chain reaction.ResultsThe results showed that the 1.5 MPa UTMD‐treated group had the highest proliferation capacity of MSCs at 24 hours. ELISA or quantitative reverse transcription polymerase chain reaction results showed that UTMD treatment of the 1.5 MPa group significantly upregulated the expression of the stemness genes NANOG, SOX‐2, and OCT‐4.ConclusionsIn conclusion, the appropriate peak PNP value of UTMD was 1.5 MPa, and 1.5 MPa‐mediated UTMD group obviously promoted MSCs proliferation and maintained stemness by upregulating the expression of stemness genes.

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“…Recently, it has been demonstrated in in vitro studies that LIPUS improves the balance between stemness and osteoblast differentiation of hMSCs by modulating different proteins such as transforming protein RhoA [3,4,5,6,7]. RhoA is a protein of the Rho Family that acts as a molecular switch responding to cell surface receptors for various cytokines, growth factors, adhesion molecules, and G-protein-coupled receptors [8,9].…”

Section: Introductionmentioning

confidence: 99%

miR-31-5p Is a LIPUS-Mechanosensitive MicroRNA that Targets HIF-1α Signaling and Cytoskeletal Proteins

Costa

Carina

Conigliaro

et al. 2019

IJMS

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The roles of low-intensity pulsed ultrasound (LIPUS) and microRNAs (miRNAs) on hMSCs commitments have already been investigated; however, the effects of the application of their co-treatments in an in vitro cell model are still unknown. Our previous studies demonstrated that (i) LIPUS modulated hMSCs cytoskeletal organization and (ii) miRNA-675-5p have a role in HIF-1α signaling modulation during hMSCs osteoblast commitment. We investigated for the first time the role of LIPUS as promoter tool for miRNA expression. Thanks to bioinformatic analysis, we identified miR-31-5p as a LIPUS-induced miRNA and investigated its role through in vitro studies of gain and loss of function. Results highlighted that LIPUS stimulation induced a hypoxia adaptive cell response, which determines a reorganization of cell membrane and cytoskeleton proteins. MiR-31-5p gain and loss of function studies, demonstrated as miR-31-5p overexpression, were able to induce hypoxic and cytoskeletal responses. Moreover, the co-treatments LIPUS and miR-31-5p inhibitor abolished the hypoxic responses including angiogenesis and the expression of Rho family proteins. MiR-31-5p was identified as a LIPUS-mechanosensitive miRNAs and may be considered a new therapeutic option to promote or abolish hypoxic response and cytoskeletal organization on hMSCs during the bone regeneration process.

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I–2 Low Intensity Pulsed Ultrasound (LIPUS) Helps to Maintain the Undifferentiated Status of Mesenchymal Stem Cells

Cited by 4 publications

References 0 publications

Osteogenic commitment and differentiation of human mesenchymal stem cells by low‐intensity pulsed ultrasound stimulation

Osteogenic commitment and differentiation of human mesenchymal stem cells by low‐intensity pulsed ultrasound stimulation

Optimal Treatment Parameters for Ultrasound‐Stimulated Microbubbles in Upregulating Proliferation and Stemness of Bone Marrow Mesenchymal Stem Cells

miR-31-5p Is a LIPUS-Mechanosensitive MicroRNA that Targets HIF-1α Signaling and Cytoskeletal Proteins

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