Instrument for stimulating adipocytes using audible sound waves

Abstract

To provide a novel method capable of noninvasively controlling the differentiation and / or accumulation of adipocytes (adipose tissue) in a living body.SOLUTION: An instrument for noninvasively applying stimulation to adipocytes in a living body of a subject, the stimulation being vibration stimulation generated by oscillation of audible sound waves, comprises audible sound wave generation means for generation of the audible sound waves.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present disclosure relates to an instrument using audible sound waves, and more particularly to an instrument for non-invasively stimulating adipocytes in a living body, an acoustic stimulation system for inhibiting adipocyte differentiation and / or accumulation in a living body, and a method for inhibiting adipocyte differentiation and / or accumulation using audible sound stimulation. [Background technology]

[0002] Currently, over one billion people worldwide are obese, and the annual medical costs of treating them amount to 590 trillion yen. It is estimated that by 2035, over half of the world's population will be obese, and the various adverse effects of obesity, such as lifestyle-related diseases, will increase dramatically. There are concerns that the current social structure will become unsustainable without effective measures to combat obesity.

[0003] The first and most important treatment for obesity is to prevent the obesity cycle by suppressing adipose tissue formation in early-stage simple obesity, which can be improved with dietary control and exercise. However, maintaining appropriate diet and exercise habits over the long term is often difficult. Furthermore, appropriate medical treatment is essential for chronic, late-stage obesity and symptomatic obesity caused by genetic or hormonal abnormalities. The only effective treatment for late-stage, chronic obesity is adipose tissue removal surgery, and the five obesity treatments previously approved by the US Food and Drug Administration (FDA) have not been widely adopted due to their high cost and limited effectiveness. In recent years, novel anti-obesity drugs have been developed that target the cellular mechanisms of fat accumulation. Ugobi, a GLP-1 receptor agonist that suppresses appetite and gastrointestinal motility, was launched in February 2024, and Arai, a lipolytic enzyme inhibitor that inhibits fat absorption, was launched in April 2024. The effectiveness of these drugs in treating obesity is expected to become clear in the future, but the former is expected to have side effects such as gastrointestinal disorders and hypoglycemia, and the latter is expected to have side effects such as digestive system failure and disorders due to inadequate intake of fat-soluble vitamins, and the choice of who can take them is expected to be limited, as it will be determined based on the patient's medical history and need for medical treatment.As such, controlling obese tissue is not easy, whether for the suppression of early-stage obesity or for the medical treatment of late-stage / chronic obesity, and the development of a simple, non-invasive, and effective method for controlling adipose tissue is desired.

[0004] As a method for reducing fat non-invasively, for example, a method for reducing fat by using ultrasound has been proposed (Patent Documents 1 and 2, etc.). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2001-286484 [Patent Document 2] Special Publication No. 2012-513837 Summary of the Invention [Problem to be solved by the invention]

[0006] Obesity has been on the rise worldwide in recent years. Obesity, particularly obesity caused by excessive body fat, can worsen lifestyle-related diseases such as diabetes and hypertension, and can even lead to arteriosclerosis, myocardial infarction, and stroke. Surgical resection, a common treatment for obesity, entails physical and mental burdens and risks, and consumes significant medical resources for surgery and postoperative care. Anti-obesity drug treatment raises concerns about its actual efficacy and side effects. Future issues include safety issues due to interactions with concomitant medications, and its versatility, including whether it is a widely applicable treatment. Therefore, there is a need for the development of new technologies that can noninvasively control the differentiation and / or accumulation of adipocytes (adipose tissue) in the body, as an alternative or complement to the current medical treatments described above.

[0007] Therefore, in one aspect, the present disclosure provides a new method capable of non-invasively controlling the differentiation and / or accumulation of adipocytes (adipose tissue) in a living body. [Means for solving the problem]

[0008] In one aspect, the present disclosure relates to an apparatus for non-invasively stimulating fat cells in a target living organism, wherein the stimulation is a vibrational stimulation generated by the oscillation of an audible sound wave, and the apparatus includes an audible sound wave generating means for generating the audible sound wave.

[0009] In another aspect, the present disclosure relates to an acoustic stimulation system for suppressing differentiation and / or accumulation of fat cells in a living organism, wherein the stimulation is a vibration stimulation generated by the oscillation of audible sound waves, and the acoustic stimulation system comprises a generating means for generating audible sound waves at least either on the water surface or underwater in a water tank capable of accommodating a human or non-human animal.

[0010] In another aspect, the present disclosure relates to a method for suppressing differentiation and / or accumulation of adipocytes in a living subject by applying vibrational stimulation to the subject through the generation of audible sound waves. [Effects of the Invention]

[0011] According to one aspect of the present disclosure, a new method capable of non-invasively controlling the differentiation and / or accumulation of adipocytes (adipose tissue) in a living body can be provided. DETAILED DESCRIPTION OF THE INVENTION

[0012] The inventor constructed a system that uses a vibrator to directly irradiate audible sound waves onto cells cultured in a culture dish, and through further research, discovered that the Ptgs2 (prostaglandin-endoperoxide synthase 2, COX-2) gene and the Ctgf (connective tissue growth factor, CCN2) gene show high and stable gene responses in different cells and under different sound wave conditions, making these genes representative of the gene responses induced by sound wave stimulation. In a previous study by the inventors, these genes were confirmed to respond when cells (e.g., mesenchymal cells, myoblasts, fibroblasts, and neuroblastoma cells) cultured in a culture dish were indirectly irradiated with sound waves using a speaker system (Masahiro Kumeta et al., "Cell type-specific suppression of mechanosensitive genes by audible sound stimulation" PLOS ONE https: / / doi.org / 10.1371 / journal.pone.0188764 January 31, 2018). Further research revealed that when sound waves are indirectly irradiated using a speaker system, not only does it vibrate the culture medium, but also the culture dish, and the vibrations generated in this dish are transmitted to the cells, suggesting that these indirectly transmitted vibrations act as repressors of the mechanosensitive genes Ptgs2 and Ctgf. Then, in the process of further analyzing the effects of audible sound wave stimulation on various cells, the inventor discovered that audible sound wave stimulation may affect the differentiation of adipocytes, and furthermore, through experiments such as gene level and fat differentiation evaluation, audible sound wave stimulation effectively suppresses the progress of differentiation and the proportion of differentiated cells. The present disclosure is based on these findings.

[0013] Although the details of the mechanism by which the effects of the present disclosure are manifested are not clear, it is presumed as follows. Prostaglandin E2 (PGE2) is known to inhibit adipocyte differentiation and promote fat breakdown via its receptor EP4. The Ptgs2 gene, which is induced by sound stimulation, is one of the factors involved in the synthesis of PGE2. When adipose tissue (adipocytes) is exposed to vibration stimulation generated by the oscillation of audible sound waves, the expression (response) of Ptgs2 is induced in the adipocytes, which increases the synthesis of PGE2, thereby suppressing the differentiation, maturation, and maintenance processes in adipocytes. As a result, it is thought that adipose tissue can be reduced and controlled. However, the present disclosure need not be construed as being limited to these mechanisms.

[0014] In one or more embodiments, the present disclosure has a technical feature of using vibration stimulation generated by the oscillation of audible sound waves, which have very little attenuation compared to ultrasound, which is used as bioacoustic stimulation. According to one or more embodiments, the vibration stimulation is transmitted to adipocytes present in subcutaneous tissue, thereby suppressing differentiation activity in the adipocytes and inducing a decrease in adipose tissue, thereby enabling simple and effective control of adipose tissue. Therefore, in one or more embodiments, the present disclosure can prevent obesity and inhibit its progression, and can also prevent and improve lifestyle-related diseases caused by obesity, maintain and manage body shape, and promote health.

[0015] In the present disclosure, "audible range" generally refers to the frequency range of sound that can be perceived by humans, and typically refers to a frequency band of 20 Hz or higher and less than 20 kHz. In one or more embodiments, the frequency of audible sound waves in the present disclosure may be 20 Hz or higher, 30 Hz or higher, 40 Hz or higher, 50 Hz or higher, 100 Hz or higher, 110 Hz or higher, 120 Hz or higher, 150 Hz or higher, or 200 Hz or higher. In one or more embodiments, the frequency of audible sound waves in the present disclosure may be less than 20 kHz, 19 kHz or lower, 18 kHz or lower, 17 kHz or lower, or 16 kHz or lower.

[0016] In the present disclosure, in one or more embodiments, "oscillation of audible sound waves" refers to converting a signal (electrical signal) in the audible frequency band into physical vibrations and generating them. In one or more embodiments, the signal (electrical signal) used is a pure tone of a single frequency in the above frequency band, and examples thereof include various waveforms such as sine waves, triangular waves, square waves, and sawtooth waves, noise sounds including any multiple frequency bands within the above frequency band, and complex sounds consisting of multiple frequency bands and varying over time. In one or more embodiments, the audible sound waves oscillated to generate a vibration stimulus in the present disclosure are preferably audible sound waves consisting of sine waves, more preferably audible sound waves consisting of a single sine wave, and even more preferably audible sound waves consisting solely of sine waves of a single frequency, from the viewpoint of being able to provide a vibration stimulus generated by oscillating a sound signal consisting exclusively of audible frequencies.

[0017] In the present disclosure, the term "audible sound wave generating means" refers to a means for generating audible sound waves, and in one or more embodiments, includes an oscillator capable of generating an audible sound signal and a vibrating unit that converts the audible sound signal from the oscillator into physical vibrations to generate sonic vibrations. In one or more embodiments, examples of the vibrating unit include a vibrating body (vibrator) and a vibrator (shaker, etc.). In one or more embodiments, the vibrating body (vibrator) can transmit (provide) vibrations by vibrating itself. In one or more embodiments, the vibrator can transmit (provide) vibrations by vibrating a member to which the vibrator is attached. In one or more embodiments, the audible sound wave generating means may include an oscillator capable of generating an audible sound signal and a vibrating body (vibrator), or may include an oscillator capable of generating an audible sound signal and a vibrator. In one or more embodiments, the audible sound wave generating means may include an oscillator capable of generating an audible sound wave signal, a vibrating body (vibrator), and a vibrator. In one or more embodiments, the audible sound wave generating means may be a speaker-type vibrator that can be used underwater. In one or more embodiments, the audible sound wave generating means may generate audible sound waves by itself, or may generate audible sound waves via a transmission means or the like, which will be described later.

[0018] In the present disclosure, "sonic stimulation" refers to a vibration stimulation generated by the oscillation of audible sound waves, and can also be called an audible sound stimulation.

[0019] In the present disclosure, "adipocytes" refer to cells that have lipid droplets in their cytoplasm and are involved in the storage and metabolism of fatty acids. In the present disclosure, "adipose tissue" refers to loose connective tissue composed of adipocytes.

[0020] Apparatus of the present disclosure In one aspect, the present disclosure relates to an instrument for non-invasively stimulating adipocytes in a living subject using audible sound waves. The instrument of the present disclosure includes a generator (audible sound wave generator) for emitting audible sound waves, and is an instrument for non-invasively applying vibrational stimulation to adipocytes in a living subject, the vibrational stimulation being generated by the generation of audible sound waves by the generator. That is, one of the technical features of the instrument of the present disclosure is the use of vibrational stimulation generated by the generation of audible sound waves as non-invasive stimulation. Thus, in one or more embodiments, the instrument of the present disclosure relates to an instrument for suppressing differentiation and / or accumulation of adipocytes in a living subject using vibrational stimulation generated by the generation of audible sound waves. In one or more embodiments, the instrument of the present disclosure can be used as an acoustic therapy instrument, although not particularly limited thereto. Audible sound waves have very little attenuation compared to the ultrasound waves used for bioacoustic stimulation, and are significantly different in frequency band and energy from ultrasound. Because the device of the present disclosure uses audible sound waves, the method of action exerted on fat cells by the device of the present disclosure is clearly different from that of general devices that use ultrasound.

[0021] In one or more embodiments, the device of the present disclosure applies a vibration stimulus generated by the oscillation of audible sound waves, which can more efficiently enhance the expression of the Ptgs2 gene and promote the synthesis of PGE2, thereby suppressing the differentiation and / or accumulation of adipocytes and promoting their decomposition, etc. Thus, in one or more embodiments, the device of the present disclosure can also be referred to as a device that enhances the expression of the Ptgs2 gene in adipocytes, or a device for promoting the synthesis of PGE2 in adipocytes.

[0022] In one or more embodiments, the sound pressure applied by the audible sound waves is 1 μPa or more and 100 kPa or less. In one or more embodiments, the sound pressure is 1 μPa or more, 10 μPa or more, 100 μPa or more, 1 mPa or more, 10 mPa or more, 100 mPa or more, 1 Pa or more, 10 Pa or more, or 50 Pa or more. In one or more embodiments, the sound pressure is 10 kPa or less, 1 kPa or less, 500 Pa or less, 400 Pa or less, or 300 Pa or less. In one or more embodiments, the sound pressure may be sound pressure in air or sound pressure in an aqueous liquid.

[0023] In one or more embodiments, the audible sound wave may be generated by continuous oscillation (PLL oscillation), FM oscillation (sweep oscillation), hyper oscillation, AM oscillation (output modulation oscillation), fixed frequency oscillation, or the like.

[0024] The vibration stimulation may, in one or more embodiments, be continuous, intermittent, or intermittent, or a combination thereof.

[0025] In one or more embodiments, the vibration stimulus may be applied directly to the subject, or may be applied indirectly to the subject via an aqueous liquid or the like.

[0026] In one or more embodiments, the device of the present disclosure includes a control unit for controlling the audible sound wave generating means. In one or more embodiments, the control unit can control the audible sound wave generating means to apply audible sound wave stimulation to the subcutaneous tissue, and preferably can control the audible sound wave generating means to generate audible sound waves so as to apply audible sound wave stimulation to the subcutaneous tissue and suppress differentiation and / or accumulation of adipocytes in the subcutaneous tissue. In one or more embodiments, the control of the audible sound wave generating means by the control unit can include controlling the frequency of the audible sound waves and / or the sound pressure applied by the audible sound waves. In one or more embodiments, the control unit may include controlling the audible sound wave generating means to generate audible sound waves of a single frequency, and from the viewpoint of being able to provide vibrational stimulation caused by the oscillation of a sound signal consisting exclusively of audible frequencies, it is preferable to control the audible sound wave generating means to generate audible sound waves composed of sine waves, more preferably to control the audible sound wave generating means to generate audible sound waves composed of one sine wave, and even more preferably to control the audible sound wave generating means to generate audible sound waves composed only of sine waves of a single frequency.

[0027] In one or more embodiments, the device of the present disclosure may include a transmission means for transmitting vibrational stimulation to a subject, which is generated by audible sound waves generated from the audible sound wave generating means. This allows the vibrational stimulation to be transmitted to the subcutaneous tissue of the subject more selectively and efficiently. In one or more embodiments, the transmission means is preferably a transmission means that can contact the subject. By including the contactable transmission means, in one or more embodiments, the vibrational stimulation can be applied with the transmission means in direct contact with the body surface of the subject, making it easier to apply the vibrational stimulation locally. In one or more embodiments, the contactable transmission means may be a carrier (housing) that can apply vibrational stimulation when attached to (in close contact with) the surface of the subject, or may be a probe that is used by contacting the surface of the subject. In one or more embodiments, the surface of the object may be a body surface such as skin (facial surface). In one or more embodiments, the transmission means may be integrated with the audible sound wave generating means (for example, the audible sound wave generating means is incorporated into a housing), or the vibration part of the audible sound wave generating means may be arranged in a housing different from the oscillation part, and may be connected to the oscillation part of the audible sound wave generating means via a cable or a wireless connection.

[0028] In one or more embodiments, the device of the present disclosure may be a device for applying vibrational stimulation to a subject via an aqueous liquid, the vibrational stimulation being generated by the oscillation of audible sound waves from an audible sound wave generating means. By generating vibrational stimulation in an aqueous liquid and applying the vibrational stimulation to a subject via the aqueous medium, it may be possible to more effectively apply vibrational stimulation to adipocytes in the subcutaneous tissue of the subject. Thus, in one or more embodiments, the device of the present disclosure may be one that generates an audible sound stimulation while the audible sound wave generating means or vibrator is immersed in the aqueous liquid, and applies the audible sound stimulation to the subject in the aqueous liquid. Alternatively, the device may be one that applies audible sound waves generated from the audible sound wave generating means or vibrator to the aqueous liquid, and then contacts the aqueous liquid with the subject, thereby applying the audible sound stimulation to the subject. In the former case, the vibrational stimulation generated by the oscillation of audible sound waves can be easily applied to the entire subject, which may enable simple suppression of overall adipocyte differentiation and / or accumulation. In the latter case, it becomes easier to locally apply vibrational stimuli generated by the emission of audible sound waves, which may enable local suppression of differentiation and / or accumulation of adipocytes. In one or more embodiments, the audible sound wave generating means in this aspect may be a speaker-type vibrator (e.g., a vibration speaker) that can be used underwater. In one or more embodiments, the device of the present disclosure may be placed in a water tank such as a bathtub. In one or more embodiments, the aqueous liquid may be water, warm water, or the like.

[0029] In one or more embodiments, the subject of the present disclosure may be a human or a non-human animal. Non-human animals are animals other than humans, and in one or more embodiments, may be mammals other than humans. In one or more embodiments, non-human animals may be monkeys, dogs, cats, chickens, rabbits, pigs, cows, horses, mice, rats, guinea pigs, hamsters, etc.

[0030] [Acoustic stimulation system] In another aspect, the present disclosure relates to an acoustic stimulation system for suppressing the differentiation and / or accumulation of adipocytes in a living organism, wherein the acoustic stimulation is a vibrational stimulation generated by the oscillation of audible sound waves, and the acoustic stimulation system includes an audible sound wave generating means for generating audible sound waves at least on the surface of or underwater in a water tank capable of accommodating a human or non-human animal. In one or more embodiments, the audible sound wave generating means in the acoustic stimulation system of the present disclosure can be an instrument of the present disclosure, an audible sound wave generating means used in the instrument of the present disclosure, or the like. The size of the water tank is not particularly limited, but in one or more embodiments, the size may be large enough to immerse all or part of a living subject (human or non-human animal). In one or more embodiments, being able to accommodate a human or non-human animal means that the part of the target living body (human or non-human animal) in which it is desired to inhibit differentiation and / or accumulation of adipocytes can be immersed.

[0031] In one or more embodiments, the acoustic stimulation system of the present disclosure includes a control means for controlling the audible sound wave generating means. In one or more embodiments, the control means can control the audible sound wave generating means to apply audible sound wave stimulation to the subcutaneous tissue, and preferably can control the audible sound wave generating means to generate audible sound waves so as to apply audible sound stimulation to the subcutaneous tissue and suppress differentiation and / or accumulation of adipocytes in the subcutaneous tissue. In one or more embodiments, the control of the audible sound wave generating means by the control means can include controlling the frequency of the audible sound waves and / or the sound pressure applied by the audible sound waves. In one or more embodiments, the control means preferably controls the audible sound wave generating means to generate audible sound waves composed of sine waves, more preferably controls the audible sound wave generating means to generate audible sound waves composed of a single sine wave, from the viewpoint of being able to provide vibrational stimulation caused by the oscillation of sound signals composed exclusively of audible frequencies, and even more preferably controls the audible sound wave generating means to generate audible sound waves composed only of sine waves of a single frequency.

[0032] [Method for inhibiting fat cell differentiation / accumulation] According to the present disclosure, by applying a vibration stimulus generated by the oscillation of audible sound waves to a living organism, it is possible to suppress the differentiation activity of adipocytes in the subject and the accumulation of adipocytes in the subject. Therefore, in another aspect, the present disclosure relates to a method for suppressing the differentiation of adipocytes in the subject by applying a vibration stimulus generated by the oscillation of audible sound waves to the living organism of the subject. In another aspect, the present disclosure relates to a method for suppressing the accumulation of adipocytes in the subject by applying a vibration stimulus generated by the oscillation of audible sound waves to the living organism of the subject. In one or more embodiments, the vibrational stimulation in the method of the present disclosure can be applied to a living organism using the device and acoustic stimulation system of the present disclosure.

[0033] In one or more embodiments, the vibration stimulation can be performed using the above-mentioned audible sound wave generating means. In one or more embodiments, the method of the present disclosure includes controlling the audible sound wave generating means to apply an audible sound wave stimulation to the subcutaneous tissue, and preferably includes controlling the audible sound wave generating means to apply an audible sound wave stimulation to the subcutaneous tissue to suppress differentiation and / or accumulation of adipocytes in the subcutaneous tissue.

[0034] In one or more embodiments, the method of the present disclosure includes controlling the frequency of audible sound waves that provide the vibrational stimulus. In one or more embodiments, the method of the present disclosure includes controlling the frequency of the audible sound waves that provide the vibrational stimulus to 20 Hz or more, 30 Hz or more, 40 Hz or more, 50 Hz or more, 100 Hz or more, 110 Hz or more, 120 Hz or more, 150 Hz or more, or 200 Hz or more, or to less than 20 kHz, 19 kHz or less, 18 kHz or less, 17 kHz or less, or 16 kHz or less.

[0035] In one or more embodiments, controlling the frequency includes using a single frequency for the audible sound waves that provide the vibrational stimulus. In one or more embodiments, the method of the present disclosure includes providing the subject with a vibrational stimulus generated by an audible sound signal of a single frequency. Thus, in one or more embodiments, the method of the present disclosure does not include providing the subject with a vibrational stimulus generated by an audible sound signal including multiple frequencies, or generating audible sound signals of different frequencies.

[0036] In one or more embodiments of the method of the present disclosure, from the viewpoint of being able to provide a vibration stimulus caused by the oscillation of a sound signal consisting exclusively of audible frequencies, it is preferable to control the audible sound wave generating means to generate audible sound waves consisting of sine waves, more preferably to control the audible sound wave generating means to generate audible sound waves consisting of a single sine wave, and even more preferably to control the audible sound wave generating means to generate audible sound waves consisting only of sine waves of a single frequency.

[0037] In one or more embodiments, the method of the present disclosure includes controlling the sound pressure applied by audible sound waves. In one or more embodiments, the method of the present disclosure includes controlling the sound pressure applied by audible sound waves to 1 μPa or more, 10 μPa or more, 100 μPa or more, 1 mPa or more, 10 mPa or more, 100 mPa or more, 1 Pa or more, 10 Pa or more, or 50 Pa or more, or 10 kPa or less, 1 kPa or less, 500 Pa or less, 400 Pa or less, or 300 Pa or less.

[0038] In one or more embodiments, the method of the present disclosure applies a vibration stimulus generated by the oscillation of a sound wave signal consisting exclusively of frequencies in the audible range, and preferably does not include applying a vibration stimulus generated by oscillations mainly composed of ultrasound to a subject.

[0039] In yet another aspect, the present disclosure relates to a method for controlling the instrument of the present disclosure, the control method comprising: a control unit for controlling an audible sound wave generating means setting a frequency of an audible sound wave so that differentiation and / or accumulation of adipocytes is suppressed; and causing the audible sound wave generating means to generate a vibration stimulus by oscillating the audible sound wave of the set frequency.

[0040] In the control method of the present disclosure, from the viewpoint of being able to provide vibration stimulation caused by the oscillation of a sound signal consisting exclusively of audible frequencies, the control unit may include controlling the audible sound wave generating means to generate audible sound waves consisting of sine waves, preferably controlling the audible sound wave generating means to generate audible sound waves consisting of one sine wave, and even more preferably controlling the audible sound wave generating means to generate audible sound waves consisting only of sine waves of a single frequency.

[0041] Hereinafter, embodiments of the present disclosure will be described with reference to the drawings. However, the present disclosure is not limited thereto.

[0042] Fig. 3 is a block diagram showing the configuration of one embodiment of the device of the present disclosure. Device 10 for providing non-invasive stimulation shown in Fig. 3 has oscillator 11 capable of generating an audible sound wave signal and vibration unit 12 connected and housed in the same housing, and vibration unit 12 converts the audible sound wave signal from oscillator 11 into vibration and outputs sonic vibration. Oscillator 11 outputs electrical vibration (vibration signal) necessary to generate only audible sound. Oscillator 11 may be connected to a signal amplifier, or oscillator 11 may have a built-in signal amplifier, as necessary. Although FIG. 3 shows only one vibration unit 12, the device 10 of the present disclosure may have two or more vibration units 12.

[0043] In Fig. 3, the oscillation unit 11 and the vibration unit 12 are arranged in one housing, but as shown in Fig. 4, the oscillation unit 11 and the vibration unit 12 may be arranged in separate housings. In this embodiment, the oscillation unit 11 and the vibration unit 12 may be connected by wire or wirelessly.

[0044] 5, the device 10 of the present disclosure may have a built-in control unit 13 for changing the frequency of the audible sound waves, the sound pressure applied by the audible sound waves, etc. By using the control unit 13 to control the vibrations caused by the oscillation of the audible sound waves output from the vibration unit 12, it is possible to more efficiently suppress the differentiation and / or accumulation of adipocytes.

[0045] 6 and 7 show examples of use of the device 10 of the present disclosure. As shown in FIG. 6, the device 10 of the present disclosure may be used by incorporating a vibration unit 12 wirelessly connected to an oscillation unit (not shown) into a housing 14 (e.g., a belt-type housing) and attaching the housing 14 to a desired position on a subject 15. Also, as shown in FIG. 7, a speaker-type housing 16 incorporating a vibration unit (not shown) may be placed in a bathtub 17 containing warm water 18. In the example shown in FIG. 7, a vibration stimulus generated by the oscillation of audible sound waves output from the vibration unit 12 can be applied to a subject (not shown) in the bathtub 17 via the warm water 18. In FIG. 7, the speaker-type housing 16 is placed on the side of the bathtub 17, but the present disclosure is not limited to this. The speaker-type housing 16 may also be placed in contact with a subject in the bathtub 17 containing warm water 18.

[0046] That is, the present disclosure may relate to one or more of the following embodiments: [1] A device for non-invasively stimulating fat cells in a living subject, The stimulus is a vibration stimulus generated by oscillation of an audible sound wave, An apparatus comprising an audible sound wave generating means for generating audible sound waves. [2] The device described in [1] for suppressing at least one of differentiation and accumulation of the fat cells by vibration stimulation generated by the oscillation of the audible sound waves. [3] The device according to [1] or [2], further comprising a transmission means capable of contacting the target and applying the vibration stimulus to the target. [4] An apparatus described in any one of [1] to [3], for oscillating the audible sound waves while the generating means is immersed in an aqueous liquid, and applying the vibration stimulus to the subject via the aqueous liquid. [5] An instrument according to any one of [1] to [4], wherein the frequency of the audible sound waves is equal to or greater than 20 Hz and less than 20 kHz. [6] The device according to any one of [1] to [5], wherein the audible sound waves are of a single frequency. [7] An apparatus described in any of [1] to [6], further comprising a control unit that controls the frequency of the audible sound waves generated from the audible sound wave generating means so as to suppress at least one of the differentiation and accumulation of fat cells. [8] An acoustic stimulation system for suppressing at least one of differentiation and accumulation of adipocytes in a living organism, comprising: The stimulus is a vibration stimulus generated by oscillation of an audible sound wave, An acoustic stimulation system comprising an audible sound wave generating means for generating audible sound waves at least on the surface of water and underwater in an aquarium capable of accommodating humans or non-human animals. [9] A method for suppressing at least one of differentiation and accumulation of fat cells in a subject by applying vibrational stimulation generated by the oscillation of audible sound waves to the subject living body. [Brief explanation of the drawings]

[0047] [Figure 1] Figure 1 shows the results of suppression of adipocyte differentiation by acoustic stimulation, and shows the relative mRNA expression levels of adipocyte differentiation marker genes Cebpa and Pparg on day 3 of differentiation of 3T3-L1 cells, as determined by real-time PCR. [Figure 2] Figure 2 shows the results of suppression of adipocyte differentiation by acoustic stimulation. Figure 2A shows stained images of adipose 3T3-L1 cells labeled with BODIPY 493 / 503 (undifferentiated cells, early differentiated cells, and late differentiated cells), and Figure 2B shows the ratios of undifferentiated cells, early differentiated cells, and late differentiated cells on day 7 of culture. [Figure 3] FIG. 3 is a block diagram illustrating one embodiment of an instrument of the present disclosure. [Figure 4] FIG. 4 is a block diagram illustrating another embodiment of the device of the present disclosure. [Figure 5] FIG. 5 is a block diagram illustrating yet another embodiment of an apparatus of the present disclosure. [Figure 6] FIG. 6 is a schematic diagram illustrating one method of using the device of the present disclosure. [Figure 7]FIG. 7 is a schematic diagram illustrating another method of using the device of the present disclosure. [Example]

[0048] The present disclosure will be described in more detail below with reference to examples, but these are merely illustrative and the present disclosure is not limited to these examples. All references cited in this disclosure are incorporated herein by reference.

[0049] [Suppression of adipocyte differentiation by ultrasonic stimulation 1] Mouse-derived preadipocyte 3T3-L1 cells were cultured in culture dishes. When the cell density reached approximately 70%, the medium was replaced with MDI differentiation medium containing 0.5 mM methylisobutylxanthine (ab120840, Abcam, Cambridge, UK), 1 μM dexamethasone (ab120743, Abcam, Cambridge, UK), and 10 μg / ml insulin (ab123768, Abcam, Cambridge, UK) to induce adipogenesis. To apply acoustic stimulation, the tip of a vibrating plate attached to a vibrator was placed in contact with the medium. Under this condition, 440 Hz sine wave acoustic stimulation was applied continuously at an intensity (sound pressure) of 100 Pa for 3 days. The effect of acoustic stimulation on adipogenesis was then assessed by measuring the expression levels of the marker genes CCAAT / enhancer-binding protein α (Cebpa) and peroxisome proliferator-activated receptor γ (Pparg) using real-time PCR. As a control, the expression levels of the above genes were measured in cells cultured in the same manner as above, except that the cells were cultured in MDI differentiation-inducing medium for 3 days without acoustic stimulation (silent condition). These results are shown in FIG.

[0050] As shown in Figure 1, the expression levels of both the Cebpa gene and the Pparg gene were reduced by approximately 80% compared to the control, indicating that audible sound stimulation suppresses adipocyte differentiation.

[0051] [Suppression of adipocyte differentiation by ultrasonic stimulation 2] 3T3-L1 cells were cultured for 3 days under the same conditions as described above in [Suppression of adipocyte differentiation by acoustic stimulation 1]. Then, they were replaced with differentiation-promoting medium containing only insulin and cultured for an additional 4 days. To evaluate intracellular lipid accumulation by fluorescent staining, the cells were incubated for 5 minutes in medium containing 0.5 mM BODIPY 493 / 503 (D3922, Invitrogen / Thermo Fisher Scientific, Waltham, MA, USA). After washing twice with phosphate-buffered saline, the cells were observed under a confocal laser scanning microscope (FV-1200, Olympus, Tokyo, Japan). Figure 2A shows the adipocyte differentiation stage as assessed by the amount of fluorescence accumulation. To assess the effect of audible acoustic stimulation, the percentage of cells at each differentiation stage was calculated and shown in Figure 2B.

[0052] Observation using a confocal laser scanning microscope revealed that the state of cell differentiation was classified into three categories based on the state of lipid accumulation in the cells: undifferentiated (almost no lipid droplets), early differentiation (lipid droplets several micrometers in diameter are present), and late differentiation (many lipid droplets are 10 micrometers or larger in diameter) (Figure 2A). Analysis of the proportion of cells incubated for 7 days based on this classification revealed that acoustic stimulation reduced the proportion of late differentiation cells by approximately 15% compared to cells in the silent condition (control), and significantly increased the proportion of undifferentiated cells by approximately 20% (Figure 2B). These findings demonstrate that audible sound stimulation can slow the progression of adipocyte differentiation and control adipocyte differentiation.

[0053] [Suppression of adipocyte differentiation by ultrasonic stimulation 3] 3T3-L1 cells were induced to differentiate under the same conditions as described above in [Suppression of adipocyte differentiation by ultrasonic stimulation 1], except that 440 Hz sine wave ultrasonic stimulation at an intensity of 100 Pa was performed periodically for 2 hours every 24 hours for 3 days (2 hours x 3 times).The expression levels of Cebpa and Pparg genes were measured after 3 days, and observations were made using BODIPY staining after 7 days. As a result, statistically significant suppression of the expression levels of both Cebpa and Pparg genes was confirmed (Fig. 1). Furthermore, the proportion of undifferentiated cells in cells exposed to periodic acoustic stimulation (2 hours x 3 times) was similar to that observed in cells exposed to continuous acoustic stimulation for 3 days (Fig. 2B). Therefore, it has become clear that periodic (intermittent) audible sound stimulation can also slow the progression of adipocyte differentiation and control adipocyte differentiation.

Claims

1. An apparatus for non-invasively stimulating fat cells in a target living body, The stimulus is a vibration stimulus generated by oscillation of an audible sound wave, An apparatus comprising: an audible sound wave generating means for generating audible sound waves.

2. The device according to claim 1, for suppressing differentiation and / or accumulation of the adipocytes by vibration stimulation caused by the oscillation of the audible sound waves.

3. The device of claim 1 , further comprising a transmission means capable of contacting the object and applying the vibration stimulus to the object.

4. 2. The device according to claim 1, wherein the generating means is immersed in an aqueous liquid and oscillates the audible sound waves to apply the vibration stimulus to the subject through the aqueous liquid.

5. The device of claim 1 , wherein the frequency of the audible sound waves is greater than or equal to 20 Hz and less than 20 kHz.

6. The device of claim 1 , wherein the audible sound waves are a single frequency.

7. The device according to any one of claims 1 to 6, further comprising a control unit that controls the frequency of the audible sound waves generated from the audible sound wave generating means so as to suppress differentiation and / or accumulation of adipocytes.

8. An acoustic stimulation system for suppressing differentiation and / or accumulation of adipocytes in a living organism, The stimulus is a vibration stimulus generated by oscillation of an audible sound wave, An acoustic stimulation system comprising an audible sound wave generating means for generating audible sound waves at least on the surface of water and underwater in an aquarium capable of accommodating humans or non-human animals.

9. A method for suppressing differentiation and / or accumulation of fat cells in a subject by applying vibrational stimulation generated by the oscillation of audible sound waves to the subject living body.

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