In vitro effects of therapeutic ultrasound on cell proliferation, protein synthesis, and cytokine production by human fibroblasts, osteoblasts, and monocytes

Doan, Ninh; Reher, Peter; Meghji, S; Harris, Malcolm

doi:10.1016/s0278-2391(99)90281-1

Cited by 281 publications

(211 citation statements)

References 52 publications

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“…Several reports have shown that LIUS can stimulate cell growth. Doan et al (21) reported that therapeutic LIUS (0.1 to 1 W/cm 2 ) induced the proliferation of fibroblasts and osteoblasts and, more recently, Harle et al described consistent findings (32). 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.…”

Section: Discussionsupporting

confidence: 58%

“…Chondrocyte proliferation was measured by 3 H-thymidine incorporation in the explant cultures (21,22). The explant cultures were treated with 10 mCi/mL of 3 H-thymidine (Amersham Biosciences, Piscataway, NJ, USA) for 24 h. The explants were rinsed 3-4 times with phosphate-buffered saline (PBS) and digested with papain (125 mg/ml in 0.1 M sodium phosphate, 5 mM Na 2 -ethylenedoaminetetraacetic acid (EDTA), and 5 mM cysteineHCl, pH 6.5) for 4-6 h at 60˚C.…”

Section: Assay For Chondrocyte Proliferationmentioning

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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Section: Discussionsupporting

confidence: 58%

Section: Assay For Chondrocyte Proliferationmentioning

confidence: 99%

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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“…It has been established that BMP-2 plays a central role in initiating bone formation [42], stimulating osteogenic differentiation of MSCs [9], and increasing osteoblast activity [10]. The lower BMP-2 expression could reduce local MSC activity and the capacity for bone repair [32].…”

Section: Discussionmentioning

confidence: 99%

Low-intensity Pulsed Ultrasound Enhances Bone Repair in a Rabbit Model of Steroid-associated Osteonecrosis

Zhu

Cai

Lin

et al. 2015

Clinical Orthopaedics &Amp; Related Research

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Background Steroids are a leading cause of femoral head osteonecrosis. Currently there are no medications available to prevent and/or treat steroid-associated osteonecrosis. Low-intensity pulsed ultrasound (LIPUS) was approved by the FDA for treating delayed union of bone fractures. Some studies have reported that LIPUS can enhance bone formation and local blood flow in an animal model of fracture healing. However, whether the effect of osteogenesis and neovascularization by LIPUS can enhance the repair progress in steroid-associated osteonecrosis is unknown.Questions/purposes We hypothesized that LIPUS may facilitate osteogenesis and neovascularization in the reparative processes of steroid-associated osteonecrosis. Using a rabbit animal model, we asked whether LIPUS affects (1) bone strength and trabecular architecture; (2) blood vessel number and diameter; and (3) BMP-2 and VEGF expression. Methods Bilateral femoral head necrosis was induced by lipopolysaccharide and methylprednisolone in 24 rabbits. The left femoral heads of rabbits received LIPUS therapy (200 mW/cm 2 ) for 20 minutes daily and were classified as the LIPUS group. The right femoral heads of the same rabbits did not receive therapy and were classified as the control group. All rabbits were euthanized 12 weeks after LIPUS therapy. Micro-CT, biomechanical testing, histologic evaluation, immunohistochemistry, quantitative realtime PCR, and Western blot were used for examination of the effects of LIPUS. Results Twelve weeks after LIPUS treatment, the loading strength in the control group was 355 ± 38 N (95% CI, 315-394 N), which was lower (p = 0.028) than that in the LIPUS group (441 ± 78 N; 95% CI, 359-524 N). The bone tissue volume density (bone volume/total volume) in the LIPUS group (49.29% ± 12.37%; 95 % CI, 36.31%-62.27%) was higher (p = 0.022) than that in the control group (37.93% ± 8.37%; 95 % CI, 29.15%-46.72%). The percentage of empty osteocyte lacunae in the LIPUS group (17% ± 4%; 95% CI, 15%-20%) was lower (p = 0.002) than that in the control group (26% ± 9%; 95% CI, 21%-32%). The mineral apposition rate (lm/day) in the LIPUS group (2.3 ± 0.8 lm/day; 95% CI, 1.8 2.8 lm/day) was higher (p = 0.001) than that in the control group (1.6 ± 0.3 lm/day; 95% CL, 1.4-1.8 lm/day). The number of blood vessels in the LIPUS group (7.8 ± 3.6/mm 2 ; 95% CI, 5.5-10.1 mm 2 ) was greater (p = 0.025) than the number in Hanxiao Zhu and Xunzi Cai contributed equally to this work and should be considered as co-first authors.The institution of the authors has received funding from the National Natural Science Foundation of China (Hangzhou, Zhejiang, China) (81101377, 81101345, 81171687, 81371954, 81201414, 81201416, 81401785, 81472113) 123Clin Orthop Relat Res (2015) 473:1830-1839 DOI 10.1007 Clinical Orthopaedics and Related Research ® A Publication of The Association of Bone and Joint Surgeons® the control group (5.7 ± 2.6/mm 2 ; 95% CI, 4.0-7.3 mm 2 ). Messenger RNA (mRNA) and protein expression of BMP-2 in the LIPUS group (75 ± 7, 95% CI, 70-7...

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“…The primary effect of LIPUS stimulation found at the early stage of consolidation of distraction osteogenesis might be associated with enhancement of intramembranous ossification in consolidation stage as reported by others, 23 suggesting that osteoblasts might play an important role in LIPUS-induced bone formation. In fact, in vitro experiments demonstrated that LIPUS exerted mechanical stress on osteogenic cells directly to stimulate proliferation 24,25 and osteoblastic activity. 26 also stimulated osteoblasts to synthesize nitric oxide and prostaglandin E 2 (PGE 2 ) 28,29 which enhanced bone formation.…”

Section: Discussionmentioning

confidence: 99%

Low intensity pulsed ultrasound accelerated bone remodeling during consolidation stage of distraction osteogenesis

Chan

Qin

Lee

et al. 2005

Journal Orthopaedic Research

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Bone regeneration in distraction osteogenesis occurs under tensile stress with axial rhythmic distraction after osteotomy. In this study, we evaluated if the low intensity pulsed ultrasound (LIPUS) was also effective on enhancement of bone remodeling during consolidation stage of distraction osteogenesis. Open osteotomy of seventeen 18-week-old female New Zealand rabbit tibiae were performed. The distraction was applied with the rate of 1 mm per day. LIPUS (30 mW/cm 2 , 1.5 MHz) was delivered for 20 min per day during 4-week consolidation stage (n ¼ 10). The animals without treatment served as sham group (n ¼ 7). Plain X-ray, peripheral quantitative computational tomography (pQCT), and torsional test were performed. Results showed that smaller radiolucent interzone of LIPUS treatment group was gradually occupied by calcified tissue in plain X-ray at week 2. The bone mineral density (BMD) measured on radiographs increased by 9.18% in the LIPUS group. Bone mineral content (BMC), hard callus volume, and bone strength index (BSI) measured by pQCT were 83%, 116%, and 94%, respectively, in LIPUS group that were significantly greater than those of the controls. At the 4th week, LIPUS-treated callus showed the development of neocorticalization in the proximal and distal region. The BMC, hard callus volume, and BSI of LIPUS group decreased and was not significantly different from control. This was also confirmed by the maximum torque of LIPUS-treated callus (1424.2 AE 457.3 N Á mm) obtained at week 4, which did not differ from that of the sham group (1968.8 AE 895.1 N Á mm). In conclusion, the effective period of LIPUS treatment was at the initial stage of consolidation, with accelerated bone formation and remodeling. ß

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In vitro effects of therapeutic ultrasound on cell proliferation, protein synthesis, and cytokine production by human fibroblasts, osteoblasts, and monocytes

Cited by 281 publications

References 52 publications

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

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

Low-intensity Pulsed Ultrasound Enhances Bone Repair in a Rabbit Model of Steroid-associated Osteonecrosis

Low intensity pulsed ultrasound accelerated bone remodeling during consolidation stage of distraction osteogenesis

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