Device for promoting healing of ulcer on body surface

Abstract

A body surface ulcer healing promoting device is provided with: a glucose concentration sensor (11) that detects the glucose concentration in the exudate of an ulcer (2) on a body surface (1); vibrators (21A, 21B, 21C) that impart vibrations to the ulcer (2); and a control device (31) that controls the vibrations of the vibrators (21A, 21B, 21C) on the basis of the glucose concentration in the exudate.

Description

Technical Field

[0001] The invention relates to a treatment technology, and particularly to a device for promoting the healing of body surface ulcers. Background Art

[0002] Diabetes and its complications are the main non-traumatic causes of lower limb amputation. In most cases, foot ulcers occur before lower limb amputation surgery. At the stage of foot ulcer development, it is usually difficult to correct hyperglycemia. It is believed that only two-thirds of foot ulcers are eventually cured, and the remaining foot ulcers result in some form of amputation surgery. Foot ulcers and amputation surgery have a significant impact on the lives of patients (for example, refer to patent documents 1 to 3). As a treatment method for foot ulcers, there is a method of local administration of platelet-derived growth factor.

[0003] Prior art literature Patent Literature Patent Document 1: Japanese Patent No. 5715326 Patent Document 2: Japanese Patent No. 5252585 Patent Document 3: Japanese Patent No. 5558573 Summary of the invention

[0004] Technical issues There are few treatment options for ulcers on the body surface, and patients are looking for a means to promote the healing of ulcers while in their daily lives. Therefore, one of the objects of the present invention is to provide a device for promoting the healing of ulcers on the body surface.

[0005] Technical Solution The device for promoting healing of a body surface ulcer according to the present invention comprises a glucose concentration sensor for detecting the glucose concentration in the exudate of the body surface ulcer, a vibrator for vibrating the ulcer, and a control device for controlling the vibration of the vibrator based on the glucose concentration in the exudate.

[0006] In the above-mentioned device for promoting healing of body surface ulcers, the ulcer may be an ulcer caused by diabetes.

[0007] In the above-mentioned device for promoting healing of body surface ulcers, the ulcer may be a foot ulcer caused by diabetes.

[0008] The above-mentioned body surface ulcer healing promoting device may include a plurality of vibrators.

[0009] In the above-mentioned body surface ulcer healing promoting device, the control device may control the vibration of the vibrator in a manner to promote the production of arginine in the cells.

[0010] In the above-mentioned body surface ulcer healing promoting device, the control device may control the vibration of the vibrator so that the glucose concentration in the exudate becomes equal to or higher than the concentration that increases the blood flow.

[0011] In the above-mentioned body surface ulcer healing promoting device, the control device may vibrate the vibrator until the glucose concentration in the exudate becomes or lower than the concentration at which the amount of arginine produced in the cells becomes equal to or higher than a threshold value.

[0012] The above-mentioned body surface ulcer healing promoting device may further include a blood flow sensor for detecting blood flow in the vicinity of the ulcer, and the control device may control the vibration of the vibrator based on the blood flow.

[0013] In the above-mentioned body surface ulcer healing promoting device, the control device can control the vibration of the vibrator in a manner to increase blood flow.

[0014] Technical Effects According to the present invention, it is possible to provide a device for promoting the healing of an ulcer on a body surface. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Schematic diagram showing a device for promoting healing of a body surface ulcer according to an embodiment.

[0016] Figure 2 Schematic diagram showing the relationship between hyperglycemia and metabolism according to an embodiment.

[0017] Figure 3 Schematic diagram showing the relationship between hyperglycemia and metabolism according to an embodiment.

[0018] Figure 4 This is a graph showing the uptake of 2-deoxyglucose in Examples.

[0019] Figure 5 This is a graph showing the uptake of 2-deoxyglucose in Examples.

[0020] Figure 6 This is a graph showing the uptake of 2-deoxyglucose in Examples.

[0021] Figure 7 This is a graph showing the uptake of 2-deoxyglucose in Examples.

[0022] Figure 8 is a schematic diagram showing GLUT4 translocation according to an example.

[0023] Fig. 9 Schematic diagrams and photographs showing GLUT4 translocation of the Examples.

[0024] Fig.10 These are photographs showing whether GLUT4 in the example is translocated.

[0025] Fig.11 This is a graph showing the ratio of the intensity of fluorescence from GLUT4 in the cell membrane to the intensity of fluorescence from GLUT4 in the cytoplasm in the Example.

[0026] Fig.12 This is a photograph of a rat in Example.

[0027] Fig.13 These are photographs of wounds on rats in Examples.

[0028] Fig.14 These are photographs of wounds on rats in Examples.

[0029] Fig.15 This is a graph showing the areas of wounds in rats of Examples.

[0030] Fig.16 This is a photograph of the tissue of the wound part of the rat in the Example.

[0031] Fig.17 This is a photograph of the tissue of the wound part of the rat in the Example.

[0032] Fig.18 This is a graph showing the blood volume of rats in Examples.

[0033] Fig.19 is a circuit diagram of the excitation device of the embodiment.

[0034] Fig. 20 These are photographs of wounds on rats in Examples.

[0035] Fig.21 This is a graph showing the areas of wounds in rats of Examples.

[0036] Fig. 22 This is a graph showing the blood volume of rats in Examples.

[0037] Fig.23 This is a photograph of the tissue of the wound part of the rat in the Example.

[0038] Fig.24 This is a graph showing the gene expression levels in rats of the Example.

[0039] Fig.25 This is a graph showing the gene expression levels in rats of the Example.

[0040] Fig.26 This is a graph showing the gene expression levels in rats of the Example.

[0041] Fig. 27 This is a photograph of the tissue of the wound area of ​​a rat stained with the anti-PTX3 antibody of the Example.

[0042] Fig.28 This is a graph showing the value obtained by dividing the number of CD68-positive cells by the number of CD163-positive cells (M1 / M2 macrophage ratio) in the Example.

[0043] Explanation of symbols 11... glucose concentration sensor, 12... blood flow sensor, 21... vibrator, 31... control device, 41... dressing. DETAILED DESCRIPTION

[0044] like Figure 1 As shown, the body surface ulcer healing promotion device of the embodiment comprises: a glucose concentration sensor 11 for detecting the glucose concentration in the exudate of the ulcer 2 on the body surface 1, vibrators 21A, 21B, 21C for applying vibration to the ulcer 2, and a control device 31 for controlling the vibration of the vibrators 21A, 21B, 21C based on the glucose concentration in the exudate.

[0045] As the vibrators 21A, 21B, and 21C, motors, pneumatic devices, transducer materials, piezoelectric foils, voice coils, and piezoelectric actuators can be used. The number of vibrators 21A, 21B, and 21C provided in the body surface ulcer healing promotion device of the embodiment is not particularly limited, and may be one or more. For example, the vibrators 21A, 21B, and 21C are provided in a manner surrounding the patient's ulcer 2. The vibrators 21A, 21B, and 21C generate low-frequency vibrations, for example.

[0046] The low frequency is, for example, 20 Hz to 100 Hz, 30 Hz to 90 Hz, 40 Hz to 80 Hz, 40 Hz to 70 Hz, or 40 Hz to 60 Hz, but is not particularly limited. The intensity of the vibration is, for example, 100 mVpp to 3000 mVpp, 500 mVpp to 2500 mVpp, 500 mVpp to 2000 mVpp, or 800 mVpp to 2000 mVpp, but is not particularly limited.

[0047] The vibrators 21A, 21B, 21C may be arranged on the body surface 1 via the dressing 41. Examples of the material of the dressing 41 include silicone, polyurethane, a hydrophilic film, a hydrophilic fiber, a hydrocolloid, a hydrogel, cellulose acetate, and polyvinyl alcohol.

[0048] The glucose concentration sensor 11 is configured to be in contact with the patient's ulcer 2, for example. The principle of the glucose concentration sensor 11 detecting the concentration of glucose is not particularly limited. For example, the glucose concentration sensor 11 includes a membrane that fixes glucose oxidase, a platinum electrode, and a silver electrode. If glucose is decomposed into gluconic acid and hydrogen peroxide by glucose oxidase, the hydrogen peroxide is oxidized at the platinum electrode and reduced at the silver electrode. Thus, the current generated at the electrode is proportional to the glucose concentration. The glucose concentration sensor 11 detects the glucose concentration in the exudate of the ulcer 2 and transmits the glucose concentration in the exudate of the ulcer 2 to the control device 31.

[0049] The control device 31 controls the vibration of the vibrators 21A, 21B, and 21C so that the vibration of the vibrators 21A, 21B, and 21C satisfies the conditions for promoting the production of arginine in cells. Examples of the conditions for promoting the production of arginine in cells include the intensity and frequency of the vibration.

[0050] The control device 31 may store the relationship between the glucose concentration in the exudate and the amount of arginine produced in the cells obtained in advance. In addition, the control device 31 may also store the threshold value of the amount of arginine produced in the cells required for the appropriate synthesis of nitric oxide obtained in advance. Generally speaking, the glucose concentration in the exudate and the amount of arginine produced in the cells are in a negative proportional relationship. Therefore, if the glucose concentration in the exudate is high, the amount of arginine produced in the cells is small, and if the glucose concentration in the exudate is low, the amount of arginine produced in the cells is large.

[0051] The control device 31 controls the vibration of the vibrators 21A, 21B, and 21C so that the glucose concentration in the exudate detected by the glucose concentration sensor 11 becomes a concentration or less at which the amount of arginine produced in the cells becomes a threshold value or more. The control device 31 can vibrate the vibrators 21A, 21B, and 21C until the glucose concentration in the exudate detected by the glucose concentration sensor 11 becomes a concentration or less at which the amount of arginine produced in the cells becomes a threshold value or more.

[0052] The control device 31 may store the relationship between the glucose concentration in the exudate and the blood flow obtained in advance. In addition, the control device 31 may also store the appropriate blood flow threshold obtained in advance. Generally, the glucose concentration in the exudate is in a negative proportional relationship with the blood flow. Therefore, if the glucose concentration in the exudate is high, the blood flow is small, and if the glucose concentration in the exudate is low, the blood flow is large.

[0053] The control device 31 controls the vibration of the vibrators 21A, 21B, and 21C so that the glucose concentration in the transudate detected by the glucose concentration sensor 11 becomes a concentration below the concentration at which the blood flow reaches a threshold value or more. The control device 31 may vibrate the vibrators 21A, 21B, and 21C until the glucose concentration in the transudate detected by the glucose concentration sensor 11 becomes a concentration below the concentration at which the blood flow reaches a threshold value or more.

[0054] The body surface ulcer healing promotion device of the embodiment may further include a blood flow sensor 12 for detecting blood flow near the ulcer. The principle of the blood flow sensor 12 detecting blood flow is not particularly limited. For example, the blood flow sensor 12 includes a light source and a light receiving element. The laser emitted from the light source to the human body is scattered by the red blood cells in the blood vessels. The blood flow sensor 12 receives the scattered light using the light receiving element and detects the blood flow based on the spectrum of the scattered light. The blood flow sensor 12 transmits the detected blood flow to the control device 31.

[0055] The control device 31 can control the vibration of the vibrators 21A, 21B, 21C based on the blood flow. For example, the control device 31 can control the vibration of the vibrators 21A, 21B, 21C to meet the condition of increased blood flow. Examples of the condition of increased blood flow include the intensity and frequency of the vibration.

[0056] The ulcer may be an ulcer caused by diabetes. The ulcer may be a foot ulcer. Normally, in the human body, glucose taken into cells from blood vessels is used for the production of ATP in the glycolytic system and the citric acid cycle. However, if the glucose taken up by cells increases due to hyperglycemia, the glucose is taken up into the polyol metabolic pathway.

[0057] like Figure 2 As shown, in the polyol metabolic pathway, glucose is converted into sorbitol by aldose reductase. If the activity of aldose reductase increases in the polyol metabolic pathway, NADPH (reduced nicotinamide adenine dinucleotide phosphate) as a coenzyme is consumed. NADPH is also used in the synthesis of nitric oxide (NO) from arginine in vascular endothelial cells, but if NADPH is consumed in the polyol metabolic pathway, the synthesis of nitric oxide decreases. In addition, if the concentration of glucose is high, the production of arginine is blocked. If the production of arginine is blocked, the synthesis of nitric oxide decreases. The decrease in nitric oxide leads to decreased blood flow and ischemia.

[0058] Without being bound by theory, it is believed that the body surface ulcer healing promotion device of the embodiment applies vibration to the ulcer 2, such as Figure 3As shown, AMPK (AMP-activated protein kinase) in the adipocytes near the ulcer 2 is activated, and the insulin resistance of the adipocytes is improved. In addition, it is believed that by improving the insulin resistance of the adipocytes, the glucose concentration near the adipocytes is made appropriate, and nitric oxide is appropriately synthesized in the vascular endothelial cells to promote blood flow. It should be noted that diabetic foot ulcers are prone to form in areas where muscles are lacking, such as metatarsal bones and heels. In addition, for diabetic patients, muscle atrophy is aggravated due to peripheral neuropathy, and muscles are lacking. Therefore, it is believed that the body surface ulcer healing promotion device of the embodiment activates AMPK in the adipocytes near the ulcer 2.

[0059] In addition, although not bound by theory, it is believed that the mechanical sensors of vascular endothelial cells respond to the mechanical stress generated by the body surface ulcer healing promotion device of the embodiment, and promote the synthesis of nitric oxide synthase (NOS) in vascular endothelial cells. In addition, it is believed that the activation of AMPK in adipocytes leads to the activation of nitric oxide synthase in vascular endothelial cells. It is believed that by promoting the synthesis and activation of nitric oxide synthase, nitric oxide is appropriately synthesized in vascular endothelial cells, thereby promoting blood flow.

[0060] The body surface ulcer healing promoting device of the embodiment can be miniaturized, so the patient can keep the body surface ulcer healing promoting device fixed to the patient's body surface while living an independent daily life. In addition, the patient's insulin resistance is not constant and may change. The body surface ulcer healing promoting device of the embodiment detects the glucose concentration in the exudate that changes due to insulin resistance, thereby controlling the vibration of the vibrators 21A, 21B, and 21C, thereby suppressing unnecessary burdens on the patient.

[0061] The stages of ulcer healing are divided into an inflammatory phase and a proliferative phase. In the inflammatory phase, neutrophils and M1 inflammatory macrophages infiltrate the ulcer and produce edema. In the proliferative phase, fibroblasts migrate from the periphery of the ulcer and reconstruct the extracellular matrix, causing angiogenesis and forming granulation tissue. In diabetic patients with chronic hyperglycemia, persistent inflammation and the inability to shift to the proliferative phase become the reason why foot ulcers are difficult to treat. The body surface ulcer healing promotion device of the embodiment locally vibrates the vicinity of the ulcer to promote the contraction of the wound as soon as possible, so that the inflammation will not persist and the ulcer will move to the proliferative phase more quickly.

[0062] (Example 1) 3T3-L1 adipocytes that had been differentiated into adipocytes for 8 days were prepared. The adipocytes were divided into a group to which vibration was applied and a group to which vibration was not applied, and the adipocytes were cultured separately. The group to which vibration was applied was subjected to 50 Hz vibration at an intensity of 600 mVpp, 800 mVpp, 1000 mVpp, 1500 mVpp, or 2000 mVpp for 40 minutes every day.

[0063] After 7 days, if Figure 4 As shown in the figure, the uptake of 2-deoxyglucose in the adipocytes of the group given vibration was greater than that of the group not given vibration. In addition, the uptake of 2-deoxyglucose in the adipocytes was examined over 7 days. The results were as follows: Figure 5 As shown, when vibration is applied, the uptake of 2-deoxyglucose by adipocytes increases over time.

[0064] The results of administration of wortmannin, an inhibitor of insulin-dependent glucose uptake pathway, to adipocytes showed that Figure 6 As shown in Figure 2, Altmanin did not significantly inhibit the effect of vibration on promoting the uptake of 2-deoxyglucose. On the other hand, when compound C, an AMPK inhibitor, was administered to adipocytes, the results were as follows: Figure 7 As shown, compound C significantly inhibited the effect of promoting 2-deoxyglucose uptake by vibration. This result suggests that the effect of promoting 2-deoxyglucose uptake by vibration is caused by the activation of AMPK by vibration.

[0065] (Example 2) GLUT4, a sugar transporter, is responsible for the intracellular uptake of blood sugar promoted by insulin and exercise. Figure 8 and Fig. 9 As shown in the figure, GLUT4 is normally stored in the GLUT4 storage compartment in the cytoplasm. If the cells are stimulated by insulin or exercise, GLUT4 is transported from the storage compartment to the cell membrane. This phenomenon, called GLUT4 translocation, is known to be inhibited by insulin resistance caused by type 2 diabetes. GLUT4 translocation can be observed by immunofluorescence staining of GLUT4.

[0066] To observe whether GLUT4 translocation occurs in 3T3-L1 adipocytes under vibration at 50 Hz, 1000 mVpp to 1500 mVpp, 40 min / day, and 5 days. Fig.10 As shown, in cells not given insulin and inhibitors, GLUT4 is present in the cytoplasm in cells not given vibration, but GLUT4 is biased to be present in the cell membrane in cells given vibration. In cells given insulin but not inhibitors, GLUT4 is biased to be present in the cell membrane in both cells not given vibration and cells given vibration. In cells given otmanin, GLUT4 is present in the cytoplasm in cells not given vibration, but GLUT4 is biased to be present in the cell membrane in cells given vibration. In cells given compound C, GLUT4 is present in the cytoplasm in cells not given vibration, and GLUT4 is also present in the cytoplasm in cells given vibration.

[0067] Fig.11 A graph showing the ratio of the intensity of fluorescence in the cell membrane to the intensity of fluorescence in the cytoplasm. In cells given compound C, no significant difference was confirmed between cells not given vibration and cells given vibration. In cells given Ultramannin, significant differences were confirmed between cells not given vibration and cells given vibration. The results also provide the following inspiration: the effect of promoting the uptake of 2-deoxyglucose caused by vibration is caused by the activation of AMPK caused by vibration.

[0068] (Example 3) Seven-week-old male SD rats were prepared, 55 mg / kg of streptozotocin was intraperitoneally administered to the rats, and rats whose blood glucose levels were 300 mg / dL or more 7 and 14 days later were selected as diabetic model rats.

[0069] like Fig.12 As shown, a full-thickness incision with a diameter of 2 cm was made on the flank of anesthetized diabetic rats, a dressing (Foamlite, ConvaTec) was applied to the wound, and the trunk was fixed with gauze. The dressing was changed every day.

[0070] Then, for 14 days, the vibrator was placed on the gauze on the wound, and low-frequency vibration was applied locally to the wound for 40 minutes while the diabetic rats were anesthetized by isoflurane inhalation and put to sleep. The oscillation period was 50 Hz, and the oscillation intensity was 0 mVpp, 300 mVpp, 600 mVpp, or 1000 mVpp.

[0071] The appearance of the wound was observed over time. Fig.13 , Fig.14 ,and Fig.15 As shown in Figure 2, the wound of the diabetic rats given vibration contracted earlier than that of the diabetic rats not given vibration. Fig.16 As shown in FIG. 1 , the wound tissue was observed by hematoxylin-eosin staining, and the results showed that the dilation of blood vessels was confirmed in the diabetic model rats given 1000 mVpp vibration compared with the diabetic model rats not given vibration. Fig.17 As shown, the dilation of blood vessels was confirmed in the diabetic model rats to which the vibration of 1000 mVpp was applied, compared with the diabetic model rats to which the vibration was not applied.

[0072] (Example 4) Non-diabetic rats and diabetic rats were anesthetized and wounds were formed in the same manner as in Example 3, and the total blood volume during anesthesia, the total blood volume during wound formation under anesthesia, and the total blood volume after awakening from anesthesia were measured using a laser tissue blood oxygen monitor (OMEGAMONITOR, BOM-L1 TRSF, OMEGAWAVE, INC.). Fig.18 As shown, in both non-diabetic rats and rats with a diabetic model, the tissue whole blood volume during awakening was greater than that during anesthesia. This result suggests that changes in tissue whole blood volume can be measured more accurately during awakening than during anesthesia.

[0073] (Example 5) Using C1034 small vibration motor (Shinshin Electric Co., Ltd.), we made Fig.19 An excitation device for a circuit equivalent to the circuit diagram shown. In the same manner as in Example 3, a full-thickness defect with a diameter of 2 cm was formed on the flank of a diabetic rat under anesthesia, a dressing was attached to the wound of the diabetic rat under anesthesia, and an excitation device was attached to the dressing. The excitation device was used to apply vibration of 50 Hz, 600 mVpp~1000 mVpp to the rat for 7 days, 40 minutes / day. When applying the vibration, the rat was not anesthetized and was kept in a conscious state. In the control group, the excitation device was attached to the rat, but the excitation device was not operated.

[0074] As a result, if Fig. 20 and Fig.21 As shown in Figure 2, the wound of the rats given vibration contracted earlier. Fig. 22 , a graph showing the blood volume of rats in the control group on day 0 after the wound was formed when the blood volume was set to 1. The blood volume of rats given vibration increased every day. Therefore, it is shown that vibration not only temporarily dilates blood vessels, but also increases the stable blood volume of rats.

[0075] In addition, 7 days after the wound was formed, Fig.23 As shown, the wound tissue was observed by hematoxylin-eosin staining, and the results showed that the diabetic rats treated with vibration had vasodilation and numerous new blood vessels compared to the diabetic rats not treated with vibration. This result indicates that vasodilation and angiogenesis lead to an increase in the steady-state blood volume of rats.

[0076] (Example 6) In Example 3, in rats given vibration, gene expression was detected on the 4th day, and the results were as follows Fig.24As shown, NOS3, a gene marker of eNOS, a NO synthase associated with vasodilation, and Vegfa, a marker of angiogenesis, increased under 1000 mVpp vibration. In addition, it is suggested that inflammation is suppressed and vascular dynamics are improved due to the reduction of Tnf-a.

[0077] In Example 3, in rats given vibration, wound healing continued on day 14, as shown in Fig.25 As shown, in rats given vibration, inflammatory markers (Tnf-a, Ptx, Ccl2) were significantly reduced compared to rats not given vibration. In addition, NOS3, which is associated with vasodilation, was significantly increased in rats given 300mVpp to 1000mVpp vibration.

[0078] In Example 5, in rats given vibration, gene expression was examined on day 7, and the results were as follows Fig.26 As shown, it was confirmed that inflammatory markers were reduced compared with the control group to which vibration was not given.

[0079] (Example 7) As in Example 3, a rat of a diabetic model was prepared, and a full-thickness defect with a diameter of 2 cm was formed on the flank of the rat of the diabetic model under anesthesia. A dressing (Foamlite, ConvaTec) was attached to the wound, and the trunk was fixed with gauze. The dressing was changed every day. Then, for 14 days, a vibrator was placed on the gauze on the wound every day, and low-frequency vibration was locally applied to the wound for 40 minutes while the rat of the diabetic model was given isoflurane inhalation anesthesia to make it fall asleep. The period of the vibration was 50 Hz, and the intensity of the vibration was 0 mVpp or 1000 mVpp.

[0080] 14 days after the wound was formed, the wound tissue was collected and immersed in a 10% formalin solution at room temperature overnight to fix the tissue. Then, the tissue was dehydrated using G-Nox (Genostaff), a substitute for ethanol and xylene, and then embedded in paraffin to prepare a paraffin block of the tissue. Furthermore, the paraffin-embedded tissue was thinly sliced ​​at a thickness of 3 μm to prepare tissue sections.

[0081] The tissue sections were immersed in G-Nox for 5 minutes × 3 times to remove paraffin from the tissue sections. Next, the tissue sections were immersed in ethanol for 5 minutes × 3 times to remove G-Nox from the tissue sections. Then, the tissue sections were washed with purified water for 5 minutes × 2 times. Furthermore, the tissue sections were incubated in 3% hydrogen peroxide diluted with methanol for 30 minutes to inactivate endogenous peroxidase in the tissue sections. In addition, the tissue sections were placed in 0.01 mol / L citrate buffer (pH 6.0) and activated by autoclaving at 121°C for 15 minutes.

[0082] Next, the tissue sections were washed with phosphate-buffered saline (PBS) and reacted with anti-penicillin 3 (PTX3) antibody (rabbit-polyclonal, 13797-1-AP, Novus Biological) diluted 100-fold with bovine serum albumin / phosphate-buffered saline overnight. Then, the anti-PTX3 antibody that did not bind to the antigen was removed from the tissue sections, and the tissue sections were washed with PBS for 5 minutes × 3 times.

[0083] In addition, the tissue sections were incubated with 1000-fold diluted horseradish peroxidase (HRP)-labeled anti-rabbit immunoglobulin antibody (Jackson ImmunoResearch) at room temperature for 1 hour. Then, the tissue sections were reacted with 0.2 mg / mL 3,3'-diaminobenzidine [DAB] (Wako Pure Chemical Industries, Ltd.) in 0.05 mol / L Tris-HCl buffer (pH 7.4) supplemented with 2% hydrogen peroxide to visualize HRP, and the reaction was stopped with pure water. Then, hematoxylin was used for contrast staining. The tissue sections were dehydrated with ethanol, made transparent with G-Nox, and then sealed with a sealing agent.

[0084] The stained tissue sections were observed under a microscope (BZ-X800, Keyence). Fig. 27 As shown in the figure, a significant decrease in PTX3, an inflammatory marker, was observed in the rats given vibration, compared with the rats not given vibration.

[0085] (Example 8) Tissue sections were prepared in the same manner as in Example 7. However, in this example, endogenous peroxidase in the tissue sections was not inactivated. The tissue sections were washed with phosphate-buffered saline (PBS), and anti-CD68 antibodies (CD68 / SR-D1 antibody (ED1), mouse-monoclonal, NB600-985-0.025, Novus Biological) and anti-CD163 antibodies (CD163, EPR19518, rabbit-monoclonal, ab182422, Abcam) diluted 100 times with bovine serum albumin / phosphate-buffered saline were reacted with the tissue sections overnight. CD68 is a marker for M1 macrophages, and M1 macrophages promote inflammatory responses. CD163 is a marker for M2 macrophages, and M2 macrophages inhibit inflammatory responses.

[0086] Then, the anti-CD68 antibody and anti-CD163 antibody that did not bind to the antigen were removed from the tissue sections, and the tissue sections were washed with PBS for 5 minutes × 3 times. Furthermore, the tissue sections were incubated with 1000-fold diluted green fluorescent dye-labeled anti-rabbit IgG antibody (Alexa Fluor 488, registered trademark, donkey, #711-545-152, Jackson ImmunoResearch) and red fluorescent dye-labeled anti-mouse IgG antibody (Alexa Fluor 594, registered trademark, donkey, #715-585-151, Jackson ImmunoResearch) at room temperature for 1 hour. Next, the tissue sections were washed with PBS, the nuclei were stained with blue fluorescent dye using DAPI, and the tissue sections were sealed.

[0087] Observe the stained tissue sections with a microscope (BZ-X800, Keyence). Cells that emit green fluorescence are CD163-positive cells. Cells that emit red fluorescence are CD68-positive cells. Calculate the number of CD68-positive cells and the number of CD163-positive cells. Furthermore, calculate the value obtained by dividing the number of CD68-positive cells by the number of CD163-positive cells (M1 / M2 macrophage ratio). The smaller the M1 / M2 ratio, the more the inflammation is improved. As a result, if Fig.28 As shown, M1 / M2 was significantly reduced in rats given vibration compared with rats not given vibration.

Claims

1. A device for promoting healing of body surface ulcers, characterized in that: have: a glucose concentration sensor that detects glucose concentration in exudate from an ulcer on a body surface; a vibrator that applies vibrations to the ulcer; and A control device controls the vibration of the vibrator based on the glucose concentration in the exudate.

2. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The ulcer is an ulcer caused by diabetes.

3. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The ulcer is a foot ulcer caused by diabetes.

4. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The body surface ulcer healing promoting device includes a plurality of the vibrators.

5. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The control device controls the vibration of the vibrator in a manner that promotes arginine production in cells.

6. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The control device controls the vibration of the vibrator so that the glucose concentration becomes equal to or higher than a concentration that increases blood flow.

7. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The control device vibrates the vibrator until the glucose concentration becomes equal to or lower than a concentration at which the amount of arginine produced in cells becomes equal to or higher than a threshold value.

8. The device for promoting healing of body surface ulcers according to claim 1, characterized in that: The body surface ulcer healing promoting device further comprises a blood flow sensor for detecting blood flow near the ulcer. The control device controls vibration of the vibrator based on the blood flow.

9. The device for promoting healing of body surface ulcers according to claim 8, characterized in that: The control device controls the vibration of the vibrator in such a manner that the blood flow is increased.

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