The pro-metabolic effects of ultrasound
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@Mauritio excellent! Exactly the lead I was looking for thanks!
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@Mauritio said in The pro-metabolic effects of ultrasound:
The contraceptive study used Intensities from 1-2.2W/cm2 .
The testosterone studies I posted use around 20mW, so the difference here is the factor 100.
surely we can't derive many meaningful conclusions from that ,besides: don't overdo it.Thanks for having a look at those studies! Yes now I notice how you carefully presented the applied mW of ultrasound power in your posts.
I take it that the medium and high settings of your device and the commonly used intensities in diagnostic, especially in Doppler ultrasound, are off the menu.
When asked, us technicians routinely don't even know how much their machines emit but it surely will be in the top of the range of the standard 720mW/cm² "ISPTA" allowance to maximize image quality (higher if the device manual considers/lays out the expected extra thermal burden).For further mW/cm² references: Diagnostic fetal ultrasound seems to be limited to 10mW/cm² in Japan. Whereas in the US the basic allowance for diagnostic fetal ultrasound is 94mW/cm², or 17mW/cm² for targeting eyes.
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@CrumblingCookie Ok good to know!
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Low-frequency (<100 kHz), low-intensity (<100 mW/cm2) ultrasound to treat venous ulcers: A human study and in vitro experiments
Here they used 20kHz or 100kHz in twenty subjects with venous ulcer wounds (not a pretty sight!), with 15 or 45 min per session for a maximum of four treatments at 100mW/cm² SPTP (note that it's periphery, no sensitive organs), with a custom, flexural transducer inside the wound dressing, a pulse repetition frequency (PRF) of 1 Hz and duty cycle of 50%. I.e. every alternating second there was an ultrasound burst. That's all very, very low frequency in the world of ultrasound.
The lower 20kHz frequency and only 15minutes treatments (∼63 J/cm2) rather than 45minutes proved effective in this very small study:"In addition, it should be noted that the average initial wound size for the five human subjects in the 20 kHz, 15 min treatment group was nearly identical to the average non-healing ulcer in the sham group (4.8 cm2 vs 4.7 cm2), showing that wounds of the same size, when treated with ultrasound did heal, whereas those not-treated did not heal.
Larger doses (i.e., those received by the 45 min at 20 kHz ultrasound or the higher intensity 200 mW/cm2in vitro group) did not show improvements in proliferation or wound healing compared to the lower (633 J) treatment dose. These findings seem to be supported by those reported by Johns (2002), whose review paper explores previously reported efficacy of different frequencies (45 kHz–3 MHz) and doses (2–150 J/cm2) and suggests that different exposure parameters may require unique energy densities or doses to reach therapeutic efficacy.
The in vitro results indicated that 20 kHz ultrasound at 100 mW/cm2 caused an average of 32% increased metabolism (p < 0.05) and 40% increased cell proliferation (p < 0.01) after 24 h when compared to the control, non-treated cells. "
The lower the ultrasound frequency the more it encroaches on LESWT / LI-ESWT (Low Energy Shock Wave Therapy / low-intensity extracorporeal shockwave therapy), which appears marvellous for healing cardiovascular systems. Typical LEWST frequencies are 120/minute, i.e. a mere 2Hz.
There's successful therapeutic overlapping between very low frequency low intensity pulsed ultrasound and LESWT e.g. in the context of diabetes and erectile dysfunction:
https://pmc.ncbi.nlm.nih.gov/articles/PMC4837316/
https://pubmed.ncbi.nlm.nih.gov/20451317/Then there's of course "bass-therapy" of listening to loud, deep, rythmic music. Which lifts depression. And I am confident could strengthen bones to the marrow similar to yet more fun than LESWT in an ortopedics office.
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The Johns 2002 paper goes into many depths:
"Specifically, ultrasound has been shown to modulate vasoconstriction; lymphocyte adhesion properties of endothelium, mast cell degranulation, phagocytosis by macrophage, production of growth factors by macrophages; calcium fluxes in fibroblasts; angiogenesis; proliferation of T-cells, osteoblasts, fibroblasts, and a number of proteins associated with inflammation and repair (IL-1, IL-2, IL-6, IL-8, interferon-γ, fibroblast growth factor-b, vascular endothelial growth factor, collagen) (Table)1,34,40–42,45–53; and to accelerate thrombolyisis.7–16
In general, most of these researchers used a frequency of 1 MHz or 3 MHz, and the intensities ranged from 0.1 to 1.5 W/cm2.
An alternative therapeutic protocol employs a frequency of 45 kHz. An intensity range of 5 to 100 mW/cm2 was shown to increase the production of IL-1, IL-8, vascular endothelial growth factor, fibroblast growth factor-b, and collagen; promote bone healing; and accelerate thrombolysis.5,45,54,55
The long-wave (45-kHz) ultrasound increases penetration depth and, therefore, seems to be more appropriate than traditional high-frequency ultrasound (1 MHz and 3 MHz) for promoting revascularization and bone healing at greater depths."
"The frequency resonance hypothesis suggests that the energy provided to the enzyme by the ultrasound wave may induce transient conformational shifts in certain enzymatic proteins, altering the enzyme's activity (ie, kinases or phosphatases) and the overall function of the cell (Figure 1). Alternatively, ultrasound's resonating force may result in the dissociation of functional multimolecular complexes (Figure 2) or the release of a sequestered molecule by dislodging an inhibitor molecule from the multimolecular complex (Figure 3). In essence, the mechanism of ultrasound's action in Figures 2 and 3 is the same. "
"Hypothetically, frequency resonance may imply that different frequencies (1 MHz, 3 MHz, 45 kHz, and others) establish unique resonant or shearing forces (or both). Moreover, various frequencies may affect combinations of proteins or multimolecular complexes in different ways, lending to the possibility of targeted effects at the cellular and molecular levels."
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@CrumblingCookie said in The pro-metabolic effects of ultrasound:
200 mW/cm2in vitro group) did not show improvements in proliferation or wound healing
Thanks for sharing ! Again it's the lower doses around 100mW/cm2 that have the best effects .
Very interesting that 200mW/cm2 didn't even work. Maybe I'm not even going to need my mid and high settings on the device. -
@CrumblingCookie said in The pro-metabolic effects of ultrasound:
There's successful therapeutic overlapping between very low frequency low intensity pulsed ultrasound and LESWT e.g. in the context of diabetes and erectile dysfunction:
Here's a study using low frequency ultrasound for erectile dysfunction. 2/Week was as effective as 3/week.
It increased erection hardness by about 25-30% .
The intensity isn't stated but based on other studies , I would guess about 0.25mW/cm2 was used.
Another benefit of this study is that it shows that ultrasound is safe to apply on the penis in reasonable amounts.
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The effect of ultrasound on aged rats seems to be different. In this study there was noe effect on sperm health or testosterone when ultrasound was applied in the following schedule :
1.5 Mhz frequency, 1 Khz
repetition pulse rate, 200 fls pulse width, 30 V peak-topeak amplitude and 20 mW/cm2 intensity.Maybe there is something to the hormesis kind of idea. Old /sick individuals also don't respond with an adequate increase in Uncoupling proteins to cold("shivering cold thermogenesis") .
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@Mauritio said:
Yes quite a lot in comparison; 300mW/cm², 5 minutes each on all four sides:
Treatment
LIPUS therapy adopts a LIPUS therapy device (WBL-ED) with a pulse duration time to pulse rest time ratio of 1:4 (200:800 ms) at 1,000 Hz and a frequency of 1.7 MHz (Wan-beili Medical Devices, Beijing, China). An F-type probe and energy level III (equivalent to 0.3 W/cm 2 ) were selected. The skin and probe were evenly coated with ultrasonic couplant. At each treatment session, LIPUS was applied on each side of the penile shaft and crus in turn, 5 minutes per site, for 20 minutes.
The response rates of the 3/W and 2/W groups increased gradually at weeks 4, 6, 8, and 12 in both the FAS and PPS, respectively. The response rates at week 12 in 3/W and 2/W groups were 62.0% and 62.5%, respectively, in the FAS. The response rates at week 12 in the 3/W and 2/W groups were 67.9% and 66.9%, respectively, in the PPS.
The percentage of patients with EHS ≥3 demonstrated significant improvements from baseline to week 12 in the 3/W group (54.9%−84.0%) and 2/W group (59.5%−83.5%),
We could hypothesize that next time they may find 2x/week with <100mW/cm² working similarly well or better not only wrt perfusion but retained/improved composition, maturation and structure of collagen (although that's a difficult thing to observe because to sacrifice and to dissect after experimentation is clearly excluded).
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I am wondering also on the impact of a very low pulse rate (as the 1Hz in the wound healing study above). Maybe that kind of emulates LESWT but I'm no biophysicist.
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Looking through the graphs in this 1999 paper doesn't definitely answer any of my thoughts but pinpoints further questions I'm having.
They used 45kHz continuous vs the usual 1MHz 1:4 pulsed ultrasound. The various protein expressions from the continous 45kHz are sometimes non-linear to the energy intensity - in contrast to the pulsed 1MHz where the dose-dependent effect appear very linear.-> Continuous US may be inferior to pulsed US. The tissues may need those resting phases between US bursts.
What's also super evident is that with the long-wave 45kHz, even after multiplying the energy intensities by x5 for equivalent joules, all the cell proliferation maxima are far lower than with 1MHz pulsed: At 5-30mW/cm² (x5 = 25-150mW/cm² pulsed) versus 100-700mW/cm².
On page 7 and 8 are the graphs for FGFb and VEGF expression from monocytes and osteoblasts and those drop off really sharply at anything but the lowest doses beyond 5 or 15mW in the 45kHz cont or beyond 100mW in the 1MHz pulsed.
FGFb (Basic fibroblast growth factor) is even strongly suppressed below control levels at more than 100mW at 1MHz pulsed.-> Variations in energy intensity, frequency and pulsing definitely induce very opposing effects on respective cellular functions. E.g. stimulating vascularization and noncollagenous protein synthesis takes place at much lower doses than the maxima for fibroblast and osteoblast proliferation.
-> Seeing, however, that even for the latter the effects by far exceeded the controls at the lowest intensities tested (5mW/cm² 45kHz cont and 100mW/cm² 1MHz pulsed) there appears to be no specific incentive to go beyond the minimum dosage in any case, based on this in vitro study.
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@CrumblingCookie said in The pro-metabolic effects of ultrasound:
LIPUS therapy adopts a LIPUS therapy device (WBL-ED) with a pulse duration time to pulse rest time ratio of 1:4 (200:800 ms) at 1,000 Hz and a frequency of 1.7 MHz (Wan-beili Medical Devices, Beijing, China). An F-type probe and energy level III (equivalent to 0.3 W/cm 2 ) were selected. The skin and probe were evenly coated with ultrasonic couplant. At each treatment session, LIPUS was applied on each side of the penile shaft and crus in turn, 5 minutes per site, for 20 minutes.
Oh i didn't catch where they mentioned the intensity.
That's way higher than I thought .
But good to know that it's safe. -
**Rejuvenation and ultrasound **
This is the most interesting study on ultrasound I've seen so far.
TLDR:
In this study low frequency ultrasound:- Increases life span and healthspan
- reverses cellular senescence, causing senescent cells to become functional again and divide
- Inhibits 15 different markers of cellular senescence (SASP)
- Drastically increases activity and physical fitness levels in ages mice by 7-10 fold
- Inhibits mTOR similarly to rapaymcin and protein restriction
- Maintains youthful appearance in old mice
- Seems to be partially Sirt1 dependent
Even if you're usually not interested in graphs, tables and such, take a look at the videos of the different groups of mice in the study. They're only 10s long.
Their appearance and movement is so different. That's almost all you need to know.
It almost looks like you see old and young mice in different videos, but they're all the same age.Ultrasound setting:
The Frequency was very low compared to what is usually used they made an in vitro test and based on that decided to go with"An LFU of 32.2 kHz at 4 kPa for 30 min with a duty cycle of 1.5 s on and 1.5 s off was optimal for inducing the growth of the senescent cells; thus, these values were used for subsequent cell and mouse experiments."
The intensity is not clear, because they bathed the mice in water while applying ultrasound. But from what I could gather from the supplementary data, it seems pretty low and in line with most studies on this thread.
Longevity and activity:
The maximal lifespan seems to be improved by 10-15%, which is quite remarkable for just sound.
The most effective ones were the groups that received the least ultrasound : every other (D2) and every third day(D3).The activity was increased drastically in ultrasound treated mice. Again D2 and D3 beeing Most effective .
"At 29 months of age, the sham mice turned the wheel only 1500 turns on average with the best performers at 2000 turns, whereas the D3 LFU-treated mice had an average of ~12,000 turns with the best at 30,000 (Figure 6c)."
The "positive bystander effect":
Interestingly, cells that were treated with ultrasound caused other cells in a senescent state to revert back to normal cells.
It's a reverse-senescene, not just in the sense that it reversed senescence, but also the opposite of what senescent cells do, which is: create more senescent cells.
So similarly to radiation damage affecting the whole body,even though only a small part was radiated, this seems to be true in a positive way for ultrasound. It benefits the whole body even though a small part was radiated."Surprisingly, ultrasound treatment of normal cells causes secretion of growth-stimulating factors that partially restore normal behavior in senescent cells..."
You could say ultrasound is not just delaying age, but partially reversing it. Normally, senescent cells don't go back to working by themselves. Their either die (apoptosis) or they keep secreting inflammatory molecules (SASP). With ultrasound neither is the case, they don't die and they don't keep secreting. They revert back to normal functional cells.
"Thus, all of these objective criteria indicate that LFU reverses senescence, and we suggest that LFU actually rejuvenates senescent cells."
Another important point is that rejuvenation seems to take place in a matter of weeks.
Here in 4 weeks the level of a senescent marker β-galactosidase went from being present in 70% of cells to 10-20%.Safety:
Seems completely safe. No tumours upon autopsy. Only benefits."In terms of the safety of LFU, half of the mice in the longevity study were treated daily with LFU for over 300 days without damage or evidence of harm from the LFU treatment. Further, the treated mice maintained a normal weight, whereas the weight of the sham mice was declining with age, and the animals that died had no tumors or obvious cause of death upon autopsy (data not shown). "
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@Mauritio Thanks, helps alot!
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Very nice find!
You shouldn't have cut off the d) and e) series of mice photographs. Or maybe you purposefully did to tease us, given the many sensations already? Aaaaw, how plump and full of hair those treated mice are in comparison to the fragile control group! And then the videos really emphasize the differences even more. Impressive. Phew.
The treated mice look kind of surreal wrt to that they evidently exhibit the body structure of aged mice but are buffed-up and full of shiny fur.The most effective ones were the groups that received the least ultrasound
That's cool!
32.2 kHz at 4 kPa for 30 min with a duty cycle of 1.5 s on and 1.5 s off
Bold and original decision and based on original in-vitro findings by the authors!
That's up my alley. Such parameters I could warm up with: Frequencies "just" above the auditory range and long pulses.
-> I've tried to find such an ultrasound device but it seems the I-Tech Mio Sonic from Italy you had found is already on the lowest spectrum and a good one in wide comparison:
Other devices I could find cost at least as much even if looking really cheap and they don't even declare their energy output but it's probably high. They are being pictured for cosmetics use with women using them around their face to treat wrinkles and rub in lotion, pointing them at their temples and sidewards to their eyes. The "professional" devices for medical practitioners costs thousands. They blast up to 3000mW/cm² with a high 150mW/cm² as their absolute bottom setting. The "LIPUS" devices are a subgroup among the devices for medical professionals and are standardized to 1.5MHz, 30mW/cm² pulsed and marketed for fracture healing (for which they are a little controversial, and otherwise on-par with LLLT).
It made me feel all nauseous, really.
It seems such a device has to be built by oneself. -
@CrumblingCookie said in The pro-metabolic effects of ultrasound:
You shouldn't have cut off the d) and e) series of mice photographs. Or maybe you purposefully did to tease us, given the many sensations already?
Lol you gotta keep em engaged, don't you ?
@CrumblingCookie said in The pro-metabolic effects of ultrasound:
Aaaaw, how plump and full of hair those treated mice are in comparison to the fragile control group! And then the videos really emphasize the differences even more. Impressive. Phew.
The treated mice look kind of surreal wrt to that they evidently exhibit the body structure of aged mice but are buffed-up and full of shiny fur.Yeah it's wild how shiny their fur is! That and the increased activity are impressive.
@CrumblingCookie said in The pro-metabolic effects of ultrasound:
Such parameters I could warm up with: Frequencies "just" above the auditory range and long pulses.
Yes and it also kinda lines up with peats recommended 50-100 kHz.
That part I cannot replicate, since my device only goes with 1MHz but at least the intensity is somewhat similar.
It's interesting that there need to be a pulsing to optimize the benefits. For once in the actual treatment but also in the sense that it should be cycled (twice a week seems optimal) .
That reminds a little of the pulsed treatment with Yamanaka factors in mice, which showed similar results.@CrumblingCookie said in The pro-metabolic effects of ultrasound:
Other devices I could find cost at least as much even if looking really cheap and they don't even declare their energy output but it's probably high. They are being pictured for cosmetics use with women using them around their face to treat wrinkles and rub in lotion, pointing them at their temples and sidewards to their eyes. The "professional" devices for medical practitioners costs thousands. They blast up to 3000mW/cm² with a high 150mW/cm² as their absolute bottom setting. The "LIPUS" devices are a subgroup among the devices for medical professionals and are standardized to 1.5MHz, 30mW/cm² pulsed and marketed for fracture healing (for which they are a little controversial, and otherwise on-par with LLLT).
It made me feel all nauseous, really.
Thanks for trying anyways.
That would've been my next step, to look for a device with lower frequency. -
@Mauritio said:
Thanks for trying anyways.
That would've been my next step, to look for a device with lower frequency."Conflicts of Interest
Authors (M.S., S.K.K., F.M., B.B.R. and R.M.) are co-authors of patents related to these studies, and M.S. and F.M. have financial interests in a company, Mechanobiologics Inc. that is planning to market LFU devices suitable for senescent cell rejuvenation in vitro and in vivo."
Researching the schematics and electrical parts list aside I am wondering how that whole-body treatment could be achieved. In the study they placed the mice on top of a plastic mesh on top of a distancing cylinder in a large beaker filled with water, so that the mice were half-submerged in water and in the far-field of the transducer, i.e. in a relatively homogenous ultrasonic field.
Simply dipping a custom-built transducer at the foot end into the bathtub wouldn't be anywhere as homogenous.
Perhaps place a super-wide-beam transducer at the bottom middle of a bathtub and put a plastic sunlounger on top to lay on. Or integrate it into one of those fancy costly floatation pods.Does anyone know how to convert those 4kPa power approximately to on-skin mW/cm²?
From looking at the supplemental figure 1 one can see that it's well possible to invest time and effort yet to no or little avail by having the parameters just crucially off: Too much pressure/power (anything above 4kPa) and there's no effect. More on- than off-time pulsing and the effects decrease. A different frequency and there are none of the wanted effects. Too little duration and there's no or little effect yet too frequent repetitions of the same and the effects dimish too.
The supplemental table S2 states the us treatment is strongly antiserotinergic: Peatarians, assemble. Whereas I don't know what to make of those strongly upregulated viral infection processes in table S1. -
I wonder if the water is not playing a role in the 29 month old mice who are showing signs of reversal of age? For instance, water can be structured many different ways, (red light, vortexing, filtering through rock like media). I wouldn’t be surprised if the ultrasound is structuring/energizing the water, and it’s the mice being bathed in this substance that is exerting the effects.
I have heals many maladies in the past simply structuring water and then soaking my feet in that water. For whatever reason that I can only guess about, my best results have always come from using sea-water, (structuring that, and then soaking my feet in that). The results are systemic. A rough patch of skin on say my knee is returned to normal by soaking my feet. An increase in energy, (not driven by cortisol) can be observed by soaking my feet.
I say all of this to say: water may be playing a key role in these findings, (though if ultrasound is capable of structuring fluid, the fluids of the body may suffice too). I just personally have had remarkable success with “bathing” in structured water.
