Low-magnitude, High-frequency Loading Research Articles

Osteogenic effect of low-intensity pulsed ultrasound and whole-body vibration on peri-implant bone. An experimental in vivo study.

The aims of this study were (i) to compare the osteogenic impact of low-intensity pulsed ultrasound (LIPUS) and low-magnitude high-frequency (LMHF) loading achieved with whole-body vibration (WBV) on peri-implant bone healing and implant osseointegration in rat tibiae, and (ii) to examine their combined effect on these processes. Titanium implants were inserted in the bilateral tibiae of 28 Wistar rats. Rats were randomly divided into four groups: LIPUS+WBV, LIPUS, WBV, and control. LIPUS was applied to the implant placement site for 20min/day on 5days/week (1.5MHz and 30mW/cm2 ). WBV was applied for 15min/day on 5days/week (50Hz and 0.5g). In the LIPUS+WBV group, both stimuli were applied under the same stimulation conditions as in the LIPUS and WBV groups. After 4weeks of treatment, peri-implant bone healing and implant osseointegration were assessed using removal torque (RT) tests, micro-CT analyses of relative gray (RG) value, and histomorphometrical analyses of bone-to-implant contact (BIC) and peri-implant bone formation (BV/TV). The LIPUS+WBV group had significantly greater BIC than the WBV and control groups. Although there were no significant intergroup differences in RT, RG value, and BV/TV, these variables tended to be greater in the LIPUS+WBV group than the other groups. The combination of LIPUS and LMHF loading may promote osteogenic activity around the implant. However, further study of the stimulation conditions of LIPUS and LMHF loading is necessary to better understand the osteogenic effects and the relationship between the two stimuli.

Effect of high-frequency loading and parathyroid hormone administration on peri-implant bone healing and osseointegration

The objective of this study is to examine the effect of low-magnitude, high-frequency (LMHF) loading, and anti-osteoporosis medications such as parathyroid hormone (PTH) and bisphosphonates on peri-implant bone healing in an osteoporosis model, and to assess their combined effects on these processes. Thirteen-week-old ovariectomized rats (n = 44) were divided into three groups: PTH, alendronate, and saline. After 3 weeks of drug administration, titanium implants were inserted into the tibiae. Each group was subdivided into two groups: with or without LMHF loading via whole-body vibration (50 Hz at 0.5 g, 15 min per day, 5 days per week). Rats were killed 4 weeks following implantation. Removal torque test, micro-CT analyses (relative gray (RG) value, water = 0, and implant = 100), and histomorphometric analyses (bone-to-implant contact (BIC) and peri-implant bone formation (bone volume/tissue volume (BV/TV))) were performed. Removal torque values and BIC were significantly differed by loading and drug administration (ANOVA). Post hoc analysis showed that PTH-treated groups were significantly higher than the other drug-treated groups. BV/TV was significantly enhanced by PTH administration. In cortical bone, RG values were significantly increased by loading. In trabecular bone, however, RG values were significantly increased by PTH administration. These findings suggest that LMHF loading and PTH can act locally and additively on the bone healing process, improving the condition of implant osseointegration.

Whole body vibration improves osseointegration by up-regulating osteoblastic activity but down-regulating osteoblast-mediated osteoclastogenesis via ERK1/2 pathway

Due to the reduction in bone mass and deterioration in bone microarchitecture, osteoporosis is an important risk factor for impairing implant osseointegration. Recently, low-magnitude, high-frequency (LMHF) vibration (LM: <1×g; HF: 20–90Hz) has been shown to exhibit anabolic, but anti-resorptive effects on skeletal homeostasis. Therefore, we hypothesized that LMHF loading, in terms of whole body vibration (WBV), may improve implant fixation under osteoporotic status. In the in vivo study, WBV treatment (magnitude: 0.3g, frequency: 40Hz, time: 30min/12h, 5days/week) was applied after hydroxyapatite-coated titanium implants were inserted in the bilateral tibiae of ovariectomized rats. The bone mass and the osteospecific gene expressions were measured at 12weeks post implantation. In the in vitro study, the cellular and molecular mechanisms underlying osteoblastic and osteoclastic activities were fully investigated using various experimental assays. Micro-CT examination showed that WBV could enhance osseointegration by improving microstructure parameters surrounding implants. WBV-regulated gene levels in favor of bone formation over resorption may be the reason for the favorable adaptive bone remolding on bone-implant surface. The in vitro study showed that vibration (magnitude: 0.3g, frequency: 40Hz, time: 30min/12h) up-regulated osteoblast differentiation, matrix synthesis and mineralization. However, mechanically regulated osteoclastic activity was mainly through the effect on osteoblastic cells producing osteoclastogenesis-associated key soluble factors, including RANKL and M-CSF. Osteoblasts were therefore the direct target cells during the mechanotransduction process. The ERK1/2 pathway was demonstrated to play an essential role in vibration-induced enhancement of bone formation and decreased bone resorption. Our data suggests that WBV was a helpful non-pharmacological intervention for improving osseointegration under osteoporosis.

Mechanical stimulation of bone marrow in situ induces bone formation in trabecular explants.

Low magnitude high frequency (LMHF) loading has been shown to have an anabolic effect on trabecular bone in vivo. However, the precise mechanical signal imposed on the bone marrow cells by LMHF loading, which induces a cellular response, remains unclear. This study investigates the influence of LMHF loading, applied using a custom designed bioreactor, on bone adaptation in an explanted trabecular bone model, which isolated the bone and marrow. Bone adaptation was investigated by performing micro CT scans pre and post experimental LMHF loading, using image registration techniques. Computational fluids dynamic models were generated using the pre-experiment scans to characterise the mechanical stimuli imposed by the loading regime prior to adaptation. Results here demonstrate a significant increase in bone formation in the LMHF loaded group compared to static controls and media flow groups. The calculated shear stress in the marrow was between 0.575 and 0.7 Pa, which is within the range of stimuli known to induce osteogenesis by bone marrow mesenchymal stem cells in vitro. Interestingly, a correlation was found between the bone formation balance (bone formation/resorption), trabecular number, trabecular spacing, mineral resorption rate, bone resorption rate and mean shear stresses. The results of this study suggest that the magnitude of the shear stresses generated due to LMHF loading in the explanted bone cores has a contributory role in the formation of trabecular bone and improvement in bone architecture parameters.

Low-magnitude high-frequency loading, by whole-body vibration, accelerates early implant osseointegration in ovariectomized rats.

Osteoporosis deteriorates jaw bone quality and may compromise early implant osseointegration and early implant loading. The influence of low-magnitude, high-frequency (LMHF) vibration on peri-implant bone healing and implant integration in osteoporotic bones remains poorly understood. LMHF loading via whole-body vibration (WBV) for 8 weeks has previously been demonstrated to significantly enhance bone-to-implant contact, peri-implant bone fraction and implant mechanical properties in osteoporotic rats. In the present study, LMHF loading by WBV was performed in osteoporotic rats, with a loading duration of 4 weeks during the early stages of bone healing. The results indicated that 4-week LMHF loading by WBV partly reversed the negative effects of osteoporosis and accelerated early peri-implant osseointegration in ovariectomized rats.

Fluid shear stress in trabecular bone marrow due to low-magnitude high-frequency vibration

Low-magnitude high-frequency (LMHF) loading has recently received attention for its anabolic effect on bone. The mechanism of transmission of the anabolic signal is not fully understood, but evidence indicates that it is not dependent on bone matrix strain. One possible source of signaling is mechanostimulation of the cells in the bone marrow. We hypothesized that the magnitude of the fluid shear stress in the marrow during LMHF loading is in the mechanostimulatory range. As such, the goal of this study was to determine the range of shear stress in the marrow during LMHF vibration. The shear stress was estimated from computational models, and its dependence on bone density, architecture, permeability, marrow viscosity, vibration amplitude and vibration frequency were examined. Three-dimensional finite element models of five trabecular bone samples from different anatomic sites were constructed, and a sinusoidal velocity profile was applied to the models. In human bone models during axial vibration at an amplitude of 1 g, more than 75% of the marrow experienced shear stress greater than 0.5Pa. In comparison, in vitro studies indicate that fluid induced shear stress in the range of 0.5 to 2.0Pa is anabolic to a variety of cells in the marrow. Shear stress at the bone-marrow interface was as high as 5.0Pa. Thus, osteoblasts and bone lining cells that are thought to reside on the endosteal surfaces may experience very high shear stress during LMHF loading. However, a more complete understanding of the location of the various cell populations in the marrow is needed to quantify the effects on specific cell types. This study suggests the shear stress within bone marrow in real trabecular architecture during LMHF vibration could provide the mechanical signal to marrow cells that leads to bone anabolism.

TRABECULAR BONE ADAPTATION TO LOW-MAGNITUDE HIGH-FREQUENCY LOADING AT MICRO-GRAVITY

Exposure to microgravity causes loss of lower body bone mass in some astronauts. Low-magnitude high-frequency loading can stimulate bone formation on earth. Here we hypothesized that low-magnitude high-frequency loading will also stimulate bone formation under microgravity conditions. Two groups of six bovine cancellous bone explants were cultured at microgravity on a Russian Foton-M3 spacecraft and were either loaded dynamically using a sinusoidal curve or experienced only a static load. Comparable reference groups were investigated at normal gravity. Bone structure was assessed by histology, and mechanical competence was quantified using mCT and FE modelling; bone remodelling was assessed by fluorescent labelling and secreted bone turnover markers. Statistical analyses on morphometric parameters and apparent stiffness did not reveal significant differences between the treatment groups. The release of bone formation marker from the groups cultured at normal gravity increased significantly from the first to the second week of the experiment by 90.4% and 82.5% in response to static and dynamic loading, respectively. Bone resorption markers decreased significantly for the groups cultured at microgravity by 7.5% and 8.0% in response to static and dynamic loading, respectively. We found low strain magnitudes to drive bone turnover when applied at high frequency, and this to be valid at normal as well as at microgravity. In conclusion, we found the effect of mechanical loading on trabecular bone to be regulated mainly by an increase of bone formation at normal gravity and by a decrease in bone resorption at microgravity. Additional studies with extended experimental time and increased samples number appear necessary for a further understanding of the anabolic potential of dynamic loading on bone quality and mechanical competence. Citation: Torcasio A, Jahn K, Van Guyse M, Spaepen P, Tami AE, et al. (2014) Trabecular Bone Adaptation to Low-Magnitude High-Frequency Loading in Microgravity. PLoS ONE 9(5): e93527. doi:10.1371/journal.pone.0093527 Editor: Ali Al-Ahmad, University Hospital of the Albert-Ludwigs-University Freiburg, Germany Received May 22, 2013; Accepted March 7, 2014; Published May 2, 2014 Copyright: 2014 Torcasio et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This study was financially supported by grant C90346 (Belgium PRODEX-9 project) and the ESA MAP grant #AO99-122. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: harry.vanlenthe@mech.kuleuven.be

Low‐magnitude high‐frequency loading via whole body vibration enhances bone‐implant osseointegration in ovariectomized rats

Osseointegration is vital to avoid long-time implants loosening after implantation surgery. This study investigated the effect of low-magnitude high-frequency (LMHF) loading via whole body vibration on bone-implant osseointegration in osteoporotic rats, and a comparison was made between LMHF vibration and alendronate on their effects. Thirty rats were ovariectomized to induce osteoporosis, and then treated with LMHF vibration (VIB) or alendronate (ALN) or a control treatment (OVX). Another 10 rats underwent sham operation to establish Sham control group. Prior to treatment, hydroxyapatite (HA)-coated titanium implants were inserted into proximal tibiae bilaterally. Both LMHF vibration and alendronate treatment lasted for 8 weeks. Histomorphometrical assess showed that both group VIB, ALN and Sham significantly increased bone-to-implant contact and peri-implant bone fraction (p < 0.05) when compared with group OVX. Nevertheless the bone-to-implant contact and peri-implant bone fraction of group VIB were inferior to group ALN and Sham (p < 0.05). Biomechanical tests also revealed similar results in maximum push out force and interfacial shear strength. Accordingly, it is concluded that LMHF loading via whole body vibration enhances bone-to-implant osseointegration in ovariectomized rats, but its effectiveness is weaker than alendronate.

Influence of whole-body vibration time on peri-implant bone healing: a histomorphometrical animal study

to examine the influence of time of low-magnitude, high-frequency (LMHF) loading, whole-body vibration (WBV) on peri-implant bone healing. a custom-made Ti implant was inserted into the medio-proximal site of one tibia of 95 rats and was left to heal for 1 or 4 weeks. The daily WBV consisted of 15 consecutive frequency steps (12, 20, 30, …, 150 Hz) at an acceleration of 0.3 g. The rats were divided into five groups with different loading times: 0 (control/non-loading), 1.25, 2.5, 5 and twice 1.25 min. (with an interim recovery period) of loading. Bone-to-implant contact (BIC) and peri-implant bone fraction were measured. BIC of every test group was significantly higher than that of the control group for both healing periods. In the 4-week healing group, BIC and BFs (in all region of interests) were significantly higher in the case of twice 1.25 min. of loading compared with 1.25 min. of loading. time of loading significantly influenced the effect of the WBV on peri-implant bone healing. Twice 1.25 min. of loading appears to have the most favourable effect. LMHF loading with a particular time sequence can stimulate peri-implant bone healing and formation.

The effect of whole-body vibration on peri-implant bone healing in rats

The aim of this study was to evaluate the effect of low-magnitude, high-frequency (LMHF) loading, applied by means of whole-body vibration (WBV), on peri-implant bone healing and implant osseointegration in rat tibiae. A custom-made titanium implant was inserted into the proximal metaphysis of the tibiae of 42 rats and left to heal for 3, 7, 14 or 25 days. Half of the animals received LMHF mechanical vibration for 5 days per week (test), whereas the others served as unloaded controls. The WBV consisted of 15 consecutive frequency steps (12, 20, 30, … to 150 Hz). Each of the 15 frequencies was applied for 2000 cycles, at an acceleration of 0.3 g. In the group with a 25-day healing period, PET images were taken at -1 (day before surgery), 3, 5, 7, 10, 14 and 21 days of loading, after an injection of [(18)F]sodium fluoride, a positron-emitting tracer. The ratio of the metabolic activity around the implants to that of a reference site (uptake ratio) was calculated as a measure of bone metabolism. Bone-to-implant contact (BIC) and peri-implant bone fraction (BF) were analysed for histomorphometrical measurement. The mean BIC and BF were significantly influenced by both the loading and the healing time (ANOVA, P<0.01). The PET images did not reveal any significant difference in uptake ratio between the test and the control implants. LMHF loading increased BIC and BF significantly. The results confirm the bone-stimulating potential of LMHF loading, through WBV, on peri-implant bone healing and osseointegration.