Mitigation of bone loss with ultrasound induced dynamic mechanical signals in an OVX induced rat model of osteopenia (original) (raw)

Low intensity ultrasound effects over osteopenic female rats bones

Acta Ortopédica Brasileira, 2003

Several studies have already shown the beneficial effects of low intensity pulsed ultrasound on osteogenesis in fracture cases. However, few reports have related the ultrasound action in bone with some injury but without fracture. Thus, we induced a rat osteopenia model by ovariectomy and the proximal third of rat femur was stimulated by ultrasound (200mus burst of 1.5 MHz sine waves repeated at 1.0 kHz, 30mW/cm², SATA) for 20 min/day, during 20 days. After the treatment period, the body weight was significantly higher in the non-treated group than the treated one. No significant difference in bone mineral content was detected among the groups (p > 0.05). Also, no significant difference was noted in the mechanical properties of the femoral diaphysis. However, histologic investigations showed that the treated femur presented less microarchitectural deterioration than the non-treated group. Moreover, it was demonstrated that the treated group did show recent bone formation which wa...

Cytokine release from osteoblasts in response to ultrasound stimulation

Biomaterials, 2003

Bone is a dynamic tissue with a well-balanced homeostasis preserved by both formation and resorption of bone. Normal turnover of bone, however, can be upset by either increased osteoclast activity or decreased osteoblast function; either mechanism alone or both may result in a net loss of bone. Both osteoclasts and osteoblasts could be stimulated by mechanical stimulation in vitro, and it is assumed that this process may occur in vivo as well. In this experiment, we investigated this hypothesis by examining the effects of ultrasound stimulation on osteoblast growth and cytokine release. With this model, we explored the mechanism of low-intensity pulsed ultrasound on osteoblasts growth and upregulation of osteoclasts formation and function by cytokine release. The results showed that specific pulsed ultrasound exposure could enhance osteoblasts population together with increase in TGFb1 secretion and decrease in concentration of IL-6 and TNFa in the culture medium. Although, animal studies and clinical trial are needed to understand the real process in the whole body, ultrasound stimulation might be a good method for prevention of bone loss due to osteoporosis. r

Effect of low-intensity pulsed ultrasound on the activity of osteoclasts: An in vitro study

Archives of Oral Biology, 2016

The objective of this in vitro study was to evaluate the effect of low-intensity pulsed ultrasound on the resorption activity of osteoclast cell cultures. Design: RAW 264.7 cells were cultured and seeded over plates that were pre-coated with a synthetic carbonate apatite, and marked with fluoresceinamine-labeled sodium chondroitin polysulfate. Plates were randomly divided into 4 groups according to the treatment assigned to each one of them: NO RANKL (no RANK-L addition and no ultrasound application), NO LIPUS (addition of RANK-L and no ultrasound application), LIPUS 10 (addition of RANK-L and 10 min of ultrasound application per day), and LIPUS 20 (addition of RANK-L and 20 min of ultrasound application per day). The ultrasound device produced 1.5 MHz pulses with a repetition rate of 1 kHz and intensity of 30 mW/cm 2. The experiment extended for one week and afterwards, resorption activity was evaluated according to the fluorescence intensity analysis and pit resorption measurements (number of pits and mean area). Results: Our experiment consistently demonstrated that low-intensity pulsed ultrasound application enhanced osteoclasts resorptive activity. In addition, it was demonstrated that when daily ultrasound application lasted longer (20 min) the resorption was the highest. Results obtained from both evaluation methods were reasonably coherent. Conclusions: Low-intensity pulsed ultrasound increases osteoclast resorptive activity in the absence of osteoblasts. This effect seems to be influenced by ultrasound treatment time. Future research might be directed to investigate osteoclast response to different ultrasound application protocols (frequencies and intensities) and potential cellular mechanisms.

Stimulation of bone repair with ultrasound: A review of the possible mechanic effects

Ultrasonics, 2014

In vivo and in vitro studies have demonstrated the positive role that ultrasound can play in the enhancement of fracture healing or in the reactivation of a failed healing process. We review the several options available for the use of ultrasound in this context, either to induce a direct physical effect (LIPUS, shock waves), to deliver bioactive molecules such as growth factors, or to transfect cells with osteogenic plasmids; with a main focus on LIPUS (or Low Intensity Pulsed Ultrasound) as it is the most widespread and studied technique. The biological response to LIPUS is complex as numerous cell types respond to this stimulus involving several pathways. Known to-date mechanotransduction pathways involved in cell responses include MAPK and other kinases signaling pathways, gap-junctional intercellular communication, up-regulation and clustering of integrins, involvement of the COX-2/PGE2, iNOS/NO pathways and activation of ATI mechanoreceptor. The mechanisms by which ultrasound can trigger these effects remain intriguing. Possible mechanisms include direct and indirect mechanical effects like acoustic radiation force, acoustic streaming, and propagation of surface waves, fluid-flow induced circulation and redistribution of nutrients, oxygen and signaling molecules. Effects caused by the transformation of acoustic wave energy into heat can usually be neglected, but heating of the transducer may have a potential impact on the stimulation in some in-vitro systems, depending on the coupling conditions. Cavitation cannot occur at the pressure levels delivered by LIPUS. In-vitro studies, although not appropriate to identify the overall biological effects, are of great interest to study specific mechanisms of action. The diversity of current experimental set-ups however renders this analysis very complex, as phenomena such as transducer heating, inhomogeneities of the sound intensity in the near field, resonances in the transmission and reflection through the culture dish walls and the formation of standing waves will greatly affect the local type and amplitude of the stimulus exerted on the cells. A future engineering challenge is therefore the design of dedicated experimental set-ups, in which the different mechanical phenomena induced by ultrasound can be controlled. This is a prerequisite to evaluate the biological effects of the different phenomena with respect to particular parameters, like intensity, frequency, or duty cycle. By relating the variations of these parameters to the induced physical effects and to the biological responses, it will become possible to derive an 'acoustic dose' and propose a quantification and cross-calibration of the different experimental systems. Improvements in bone healing management will probably also come from a combination of ultrasound with a 'biologic' components, e.g. growth factors, scaffolds, gene therapies, or drug delivery vehicles, the effects of which being potentiated by the ultrasound.

Low-Intensity Pulsed Ultrasound Produced an Increase of Osteogenic Genes Expression During the Process of Bone Healing in Rats

Ultrasound in Medicine and Biology, 2010

The aim of this study was to measure the temporal expression of osteogenic genes during the process of bone healing in low-intensity pulsed ultrasound (LIPUS) treated bone defects by means of histopathologic and real-time polymerase chain reaction (PCR) analysis. Animals were randomly distributed into two groups (n 5 30): control group (bone defect without treatment) and LIPUS treated (bone defect treated with LIPUS). On days 7, 13 and 25 postinjury, 10 rats per group were sacrificed. Rats were treated with a 30 mW/cm 2 LIPUS. The results pointed out intense new bone formation surrounded by highly vascularized connective tissue presenting a slight osteogenic activity, with primary bone deposition was observed in the group exposed to LIPUS in the intermediary (13 days) and late stages of repair (25 days) in the treated animals. In addition, quantitative realtime polymerase chain reaction (RT-qPCR) showed an upregulation of bone morphogenetic protein 4 (BMP4), osteocalcin and Runx2 genes 7 days after the surgery. In the intermediary period, there was no increase in the expression. The expression of alkaline phosphatase, BMP4 and Runx2 was significantly increased at the last period. Our results indicate that LIPUS therapy improves bone repair in rats and upregulated osteogenic genes, mainly at the late stages of recovery. (E-mail: a.renno@unifesp.br) Ó