Effects of low‐intensity pulsed ultrasound on proliferation and chondroitin sulfate synthesis of cultured chondrocytes embedded in Atelocollagen® gel

Nishikori, Tetsuya; Ochi, Mitsuo; Uchio, Yuji; Maniwa, Sokichi; Kataoka, Hiroko; Kawasaki, Kenzo; Katsube, Ken‐ichi; Kuriwaka, Masakazu

doi:10.1002/jbm.1226

Cited by 83 publications

(50 citation statements)

References 33 publications

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“…The LIUS treatment was shown to have an effect on the proliferation of fibroblsts, osteoblast, and monocyte (28,31), but not in chondrocytes both in 3-D and monolayer cultures (18,19). In our experiment, however, the pretreatment of LIUS reduced the time to population doubling of the replated cells in monolayer culture until passage 2, when compared to the untreated control (Table 2).…”

Section: Sox-9contrasting

confidence: 50%

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Low‐intensity Ultrasound Stimulation Enhances Chondrogenic Differentiation in Alginate Culture of Mesenchymal Stem Cells

et al. 2006

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Mesenchymal stem cells (MSCs) are regarded as a potential autologous source for cartilage repair, because they can differentiate into chondrocytes by transforming growth factor‐beta (TGF‐β) treatment under the 3‐dimensional (3‐D) culture condition. However, more efficient and versatile methods for chondrogenic differentiation of MSCs are still in demand for its clinical application. Recently, low‐intensity ultrasound (LIUS) was shown to enhance fracture healing in vitro and induce chondrogenesis of MSCs in vitro. In this study, we investigated the effects of LIUS on the chondrogenesis of rabbit MSCs (rMSCs) in a 3‐D alginate culture and on the maintenance of chondrogenic phenotypes after replating them on a monolayer culture. The LIUS treatment of rMSCs increased: (i) the matrix formation; (ii) the expression of chondrogenic markers such as collagen type II, aggrecan, and Sox‐9; (iii) the expression of tissue inhibitor of metalloprotease‐2 implicated in the integrity of cartilage matrix; and (iv) the capacity to maintain the chondrogenic phenotypes in a monolayer culture. Notably, LIUS effects were clearly shown even without TGF‐β treatment. These results suggest that LIUS treatment could be an efficient and cost‐effective method to induce chondrogenic differentiation of MSCs in vitro for cartilage tissue engineering.

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Section: Sox-9contrasting

confidence: 50%

“…LIUS is also reported to enhance repair of articular cartilage in animal model of cartilage defect (16,17). In cultured chondrocytes in vitro, LIUS stimulation enhances the synthesis of matrix proteins such as collagen type II and proteoglycans depending on the culture condition and LIUS intensity (18)(19)(20)(21)(22).…”

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confidence: 99%

Low‐intensity Ultrasound Stimulation Enhances Chondrogenic Differentiation in Alginate Culture of Mesenchymal Stem Cells

et al. 2006

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“…Similarly, the LIUS treatment at low intensities (50 to 120 mW/cm 2 ) did not stimulate chondrocyte proliferation in studies using a monolayer culture (12), and in atelocollagen gel culture (27). On the contrary, Huang et al found that the total cell density in osteoarthritic rat cartilage was increased after LIUS treatment as compared with the control group (11,27). Therefore, the effects of LIUS on chondrocyte proliferation are still controversial, depending on the experimental systems applied.…”

Section: Discussionmentioning

confidence: 94%

“…In this study, we selected randomly the non-affected area of human OA cartilages for explant cultures and found no significant changes in chondrocyte proliferation by various the intensity of LIUS. Similarly, the LIUS treatment at low intensities (50 to 120 mW/cm 2 ) did not stimulate chondrocyte proliferation in studies using a monolayer culture (12), and in atelocollagen gel culture (27). On the contrary, Huang et al found that the total cell density in osteoarthritic rat cartilage was increased after LIUS treatment as compared with the control group (11,27).…”

Section: Discussionmentioning

confidence: 97%

“…Histological analyses of the effects of LIUS on articular cartilage in experimental arthritis have drawn attention to the possibility that LIUS might promote arthritic cartilage repair (7,11,27). LIUS was also shown to stimulate the expression of proteoglycan and type II collagen, in a rat fracture model (13) and in a monolayer culture (10,12).…”

Section: Discussionmentioning

confidence: 99%

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Effects of low‐intensity ultrasound (LIUS) stimulation on human cartilage explants

Min

Woo²,

Cho

et al. 2006

Scandinavian Journal of Rheumatology

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Objective: To evaluate the effects of low-intensity ultrasound (LIUS) stimulation on the anabolic state of human cartilage from patients with osteoarthritis (OA). Methods: Explant cultures of human OA cartilage were stimulated for 10 min every day for 7 consecutive days using continuous-wave sonication at a frequency of 1 MHz with spatial and temporal average intensities of 0 (control), 40, 200, 500, or 700 mW/cm 2 . The effects of LIUS on cell proliferation were evaluated by 3 H-thymidine incorporation. Proteoglycan synthesis was evaluated by the incorporation of 35 S-sulfate and by Safaranin O staining. Collagen synthesis was evaluated by 3 H-proline incorporation and immunohistochemistry. Results: At an intensity of 200 mW/cm 2 , LIUS treatment induced the expression of collagen type II and proteoglycan measured by the incorporation of radioactivity and specific staining of the cartilage explants. However, the expression decreased again at the higher intensities of 500 or 700 mW/cm 2 . Ultrasound had no stimulatory effect on cell proliferation at any intensity. Conclusion: LIUS has anabolic effects on human cartilage in explant cultures, indicating a potentially important method for the repair of osteoarthritic cartilage.Osteoarthritis (OA) is the most common joint disease and is characterized by progressive joint destruction, ultimately leading to the need to replace major joints. Although the precise pathological mechanism has not yet been defined, changes in the function of the chondrocytes are thought to be the main cause of the pathology of OA, particularly the imbalance between the anabolism and the catabolism of the extracellular matrix produced from chondrocytes. Methods to increase synthesis of the cartilage matrix and inhibit its degradation may therefore provide effective treatments for OA.Ultrasound (US) has been used extensively in diagnostic applications (1, 2) and there has recently been increasing interest in its therapeutic use for various musculoskeletal disorders. Several studies have shown that low-intensity US (LIUS) is effective in regenerating injured tissues, including muscles (3, 4) and bones (5-9). In particular, LIUS has been shown to enhance the healing of fractured bones by inducing chondrocyte proliferation (7,9). When applied to a monolayer culture of human chondrocytes, LIUS also increased cellular proliferation and matrix synthesis (10). In animal studies, LIUS stimulation in both bone fractures and cartilage resulted in increased expression of proteoglycan or type II collagen rather than that of bone formation factors such as transforming growth factor-b (TGF-b), osteocalcin, alkaline phosphatase, and a1(I)-procollagen, hence protecting the collagen matrix and preventing the destruction or degradation of cartilage (11-13). However, the effects of LIUS treatment on chondrocyte proliferation and the expression of matrix proteins such as proteoglycan and collagen type II have not been clearly shown in intact human cartilage.In this study, we evaluated the LIUS effects on the ana...

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Low‐Intensity Ultrasound Upregulates the Expression of Cyclin‐D1 and Promotes Cellular Proliferation in Human Mesenchymal Stem Cells

Budhiraja

Sahu

Subramanian

2018

Biotechnology Journal

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Human mesenchymal stem cells (hMSCs) hold great potential for cellular based therapeutics and tissue engineering applications and their expansion is an interesting prospect due to their low availability from in vivo sources. Therefore, this study investigated the effect of continuous-wave low-intensity ultrasound (LIUS) at 5.0-MHz and 14.0-kPa (<20 mW cm ) on the proliferative capacity, colony-formation efficiency, genetic stability, and differentiation potential of hMSCs. Additionally, potential signaling pathways involved in LIUS-mediated proliferation of hMSCs are studied. Compared to non-stimulated controls, LIUS-treated hMSCs shows a 1.9-fold greater colony-forming efficiency and 2.5-fold higher rate of cell proliferation, respectively. Differential staining and qRT-PCR analysis for selective chondrogenic, osteogenic, and adipogenic markers further confirmed that the LIUS treatment did not impact the multipotency of hMSCs. LIUS-treated hMSCs expressed normal male karyotype. The synthesis of cyclin-D1, a master regulator of cellular proliferation, is upregulated under LIUS and its enhanced mRNA expression under LIUS is noted to be mediated by the activation of both MAPK/ERK and PI3K/AKT pathways. In conclusion, LIUS promotes proliferation and self-renewal capacity of hMSCs.

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Effects of low‐intensity pulsed ultrasound on proliferation and chondroitin sulfate synthesis of cultured chondrocytes embedded in Atelocollagen® gel

Cited by 83 publications

References 33 publications

Low‐intensity Ultrasound Stimulation Enhances Chondrogenic Differentiation in Alginate Culture of Mesenchymal Stem Cells

Low‐intensity Ultrasound Stimulation Enhances Chondrogenic Differentiation in Alginate Culture of Mesenchymal Stem Cells

Effects of low‐intensity ultrasound (LIUS) stimulation on human cartilage explants

Low‐Intensity Ultrasound Upregulates the Expression of Cyclin‐D1 and Promotes Cellular Proliferation in Human Mesenchymal Stem Cells

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