Living skin optical transparency reagent and reversible skin optical transparency method

Abstract

The invention discloses a living skin light transparentizing reagent and a reversible skin light transparentizing method, and belongs to the technical field of biomedical optical imaging. The living skin optical transparency reagent comprises a first solution and hydrogel containing food coloring, wherein the first solution is prepared from the following components in percentage by volume: 40 to 60 percent of carbohydrates, 1 to 10 percent of penetration enhancer and 35 to 55 percent of alcohol substances; the hydrogel containing the food coloring is prepared by dissolving a high-molecular compound in water and then adding the food coloring, and according to the mass percent, the content of the high-molecular compound is 0.1-1%, the content of the water is 49-89.9%, and the content of the food coloring is 10-50%. The living body skin optical transparency reagent enables skin tissues to become transparent to light, improves imaging depth, can accurately obtain distribution information of skin microvascular structures, blood flow and blood oxygen, and can reduce inflammatory response, so that the living body skin optical transparency reagent has a good application prospect in skin optical imaging.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical optical imaging, and in particular relates to a living skin light transparency reagent and a reversible skin light transparency method. Background Art

[0002] The skin is the first line of defense for organisms against external invasions and is also an important target for in vivo optical imaging research. It is of great significance in the research of peripheral vascular diseases, diabetes, tumors, and immunology. Optical imaging technology can dynamically observe the changes in skin structure, vascular function, and immune cell function during the development of the disease, thereby providing assistance for the diagnosis and treatment of skin lesions. However, the turbidity of the skin limits the penetration depth of light in the tissue, so that optical imaging technology can only obtain information on the surface of the skin, but cannot obtain the deep structure of the skin tissue, and it also severely limits the quality of optical imaging.

[0003] At present, in order to achieve high-resolution optical imaging of the skin at the living level, researchers have developed a variety of skin window models based on surgical operations. However, the establishment of surgical skin windows will inevitably cause local or systemic inflammatory responses in the body, resulting in the inability to obtain the structural and functional changes of the skin in the real microenvironment.

[0004] Therefore, it is necessary to provide a new living skin phototransparency agent and a reversible skin phototransparency method to solve the deficiencies in the prior art. Summary of the invention

[0005] The purpose of the present invention is to provide a living skin light transparent reagent and a reversible skin light transparent method. The living skin light transparent reagent provided by the present invention can make the refractive index inside the skin tissue uniform and reduce tissue scattering, making the skin tissue more transparent to light, increasing the imaging depth, and obtaining the distribution information of the skin microvascular structure, blood flow and blood oxygen.

[0006] In a first aspect, the present invention provides a living skin phototransparent reagent, which includes a first solution and a hydrogel containing edible pigments; wherein, in terms of volume percentage, the first solution includes 40-60% of a carbohydrate substance, 1-10% of a penetration enhancer and 35-55% of an alcohol substance; the hydrogel containing edible pigments is prepared by dissolving a polymer compound in water and then adding the edible pigments, wherein in terms of mass percentage, the content of the polymer compound is 0.1-1%, the content of water is 49-89.9%, and the content of the edible pigments is 10-50%.

[0007] In the present invention, the inventors have found that when the living skin light-transparent reagent is applied to the skin of a subject, the first solution penetrates into the skin tissue, replaces the water molecules in the collagen hydration layer, and dissociates the collagen of the skin tissue, thereby reducing the skin's refractive index; the hydrogel containing edible pigments further reduces the refractive index difference between water and lipids inside the skin, achieves uniformity of the refractive index inside the tissue, reduces tissue scattering, and thus makes the skin tissue transparent to light, improves imaging depth, and can accurately obtain information on the distribution of skin microvascular structure, blood flow, and blood oxygen.

[0008] It is understandable that in the preparation process of the hydrogel containing edible pigments, conventional techniques can be used, for example, the polymer compound can be directly dissolved in water, or heated to assist dissolution, and a cross-linking agent can be added to quickly obtain the hydrogel.

[0009] In some embodiments, in the first solution, the saccharide substance is selected from at least one of monosaccharides and their derivatives or polysaccharides and their derivatives.

[0010] In some embodiments, the monosaccharide and its derivatives are selected from at least one of fructose, glucose, galactose or mannose; the polysaccharide and its derivatives are selected from at least one of sucrose, lactose or maltose.

[0011] In the present invention, the inventors further discovered that carbohydrate substances have a higher refractive index and carry hydroxyl functional groups, which can interact with the dermal collagen of the skin through hydrogen bonds, thereby displacing water molecules in the collagen hydration layer, making the refractive index of each component inside the skin uniform, so as to achieve optical transparency of the skin.

[0012] It is understandable that monosaccharides and their derivatives and polysaccharides and their derivatives can be routinely selected according to actual use needs. For example, in the present invention, monosaccharides and their derivatives are preferably at least one of fructose, glucose, galactose or mannose; polysaccharides and their derivatives are preferably at least one of sucrose, lactose or maltose.

[0013] In some embodiments, in the first solution, the penetration enhancer is selected from at least one of thiazone, azone, or acetone.

[0014] In the present invention, the inventors further discovered that penetration enhancers can effectively reduce the barrier effect of the stratum corneum and dense epidermis on the penetration of reagents, thereby promoting the effective penetration of active ingredients into the skin tissue, and are mainly used to increase the penetration of reagents.

[0015] It is understandable that the penetration enhancer can be conventionally selected according to actual use needs. For example, in the present invention, the penetration enhancer is preferably at least one of thiophene, azone or acetone. Further, the penetration enhancer can be processed into a liquid state, such as heating and melting to obtain a liquid to mix with other components in the first solution.

[0016] In some embodiments, in the first solution, the alcohol substance is selected from at least one of polyethylene glycol, polypropylene glycol, or glycerol.

[0017] In the present invention, the inventors further studied and found that alcohol substances have a higher refractive index and carry hydroxyl functional groups, which can interact with the dermal collagen of the skin through hydrogen bonds, thereby replacing water molecules in the collagen hydration layer; in addition, the hydroxyl functional group can form hydrogen bonds with the collagen in the skin to achieve collagen dissociation, reduce the scattering of collagen, and homogenize the refractive index of each component inside the skin to achieve optical transparency of the skin.

[0018] It is understandable that the alcohol substance can be conventionally selected according to actual use needs. For example, in the present invention, the alcohol substance is preferably at least one of polyethylene glycol, polypropylene glycol or glycerol.

[0019] In some embodiments, in the preparation process of the hydrogel containing edible pigments, the polymer compound is selected from at least one of low melting point agarose, polyacrylamide, sodium polyacrylate or sodium hyaluronate.

[0020] In the present invention, the inventors further discovered that the polymer substance forms a hydrogel after being dissolved in water, and the hydrogel is maintained on the surface of the skin to ensure that the skin absorbs the edible pigment contained in the hydrogel.

[0021] It is understandable that the polymer material can be routinely selected according to actual use needs. For example, in the present invention, the polymer compound is preferably at least one of low melting point agarose, polyacrylamide, sodium polyacrylate or sodium hyaluronate.

[0022] In some embodiments, in the preparation of the hydrogel containing food coloring, the food coloring is selected from at least one of acid red, acid orange, acid brown, lemon yellow or sunset yellow.

[0023] In the present invention, the inventors further discovered that edible pigments have strong absorption properties and can effectively increase the refractive index of the aqueous solution after being dissolved in water. After the skin absorbs edible pigments, the refractive index difference between water and lipids inside the skin can be significantly reduced, thereby homogenizing the refractive index of each component inside the skin to achieve optical transparency of the skin.

[0024] It is understandable that the food coloring can be conventionally selected according to actual use needs. For example, in the present invention, the food coloring is preferably at least one of acid red, acid orange, acid brown, lemon yellow or sunset yellow.

[0025] In a second aspect, the present invention provides a method for reversible skin phototransparency using any of the above-mentioned living skin phototransparency reagents, comprising the following steps: S1, performing hair removal treatment on the subject's skin, and obtaining skin in an original turbid state after washing and drying; S2, dripping a first solution onto the skin in an original turbid state, applying it evenly, performing a first reaction, and then removing the first solution to obtain pretreated skin; S3, applying a hydrogel containing edible pigments onto the pretreated skin, applying it evenly, and performing a second reaction to obtain phototransparent skin.

[0026] In some embodiments, the step of removing the hydrogel containing the food coloring on the photocleared skin and then scrubbing the skin to allow the skin tissue to regenerate and return to its original turbid state is also included.

[0027] In some embodiments, in step S2, the time for the first reaction is 5-10 min; in step S3, the time for the second reaction is 8-15 min.

[0028] In some embodiments, the subject comprises a mouse.

[0029] In the present invention, by successively dripping a first solution and applying a hydrogel containing edible pigments to the skin of a test subject, the refractive index inside the skin tissue is homogenized, tissue scattering is reduced, and the skin tissue becomes transparent to light, so as to achieve high-resolution imaging of the skin vascular structure, thereby overcoming the problems of low imaging quality and shallow depth due to the high scattering characteristics of the skin; further, after becoming transparent, washing with physiological saline can renature the skin tissue and restore it to its original turbid state, thereby achieving reversible light transparency of the skin; the method is simple to operate, has a high degree of transparency, high imaging quality, and good biocompatibility.

[0030] The beneficial effects of the present invention are as follows: Different from the prior art, the present invention provides a living skin light-transparent reagent. When the living skin light-transparent reagent is applied to the skin of a subject, the first solution penetrates into the skin tissue, replaces the water molecules in the collagen hydration layer, and dissociates the collagen of the skin tissue, thereby reducing the refractive index of the skin; the hydrogel containing edible pigments further reduces the refractive index difference between water and lipids inside the skin, achieves homogenization of the refractive index inside the tissue, and reduces tissue scattering, thereby making the skin tissue transparent to light, improving the imaging depth, and being able to accurately obtain the distribution information of the skin microvascular structure, blood flow and blood oxygen, while reducing the inflammatory response of the subject. Therefore, it has a good application prospect in skin optical imaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a flow chart of the method for reversible skin phototransparency using a living skin phototransparency agent in the present invention; Figure 2The effect diagram of the phototransparency of the mouse abdominal skin tissue in Example 1 of the present invention is shown in FIG. 1 ; wherein (a) is the effect diagram before the transparency, (b) is the effect diagram after the transparency, and (c) is the effect diagram after the refolding; Figure 3 The effect diagram of the phototransparency of the living skin tissue on the back skin of the mouse in Example 2 of the present invention; wherein (a) is the effect diagram before the transparency, and (b) is the effect diagram after the transparency; Figure 4 The effect diagram of the phototransparency of the living skin tissue of the mouse leg skin in Example 3 of the present invention is shown; wherein (a) is the effect diagram before the transparency, and (b) is the effect diagram after the transparency; Figure 5 The effect diagram of the phototransparency of the mouse scalp in vivo skin tissue in Example 4 of the present invention is shown in FIG. 1 , wherein (a) is the effect diagram before the transparency, and (b) is the effect diagram after the transparency; Figure 6 The effect diagram of the phototransparency of the living skin tissue on the back skin of the mouse in Comparative Example 1 of the present invention is shown in FIG. 1 , wherein (a) is the effect diagram before the transparency, and (b) is the effect diagram after the transparency; Figure 7 The figures are the effect diagrams of the phototransparency of the living skin tissue on the back skin of mice in comparative example 2 of the present invention; wherein (a) is the effect diagram before the transparency, and (b) is the effect diagram after the transparency. DETAILED DESCRIPTION

[0032] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] The experimental methods without specific conditions in the examples are usually carried out according to conventional conditions and conditions described in the manual, or according to conditions recommended by the manufacturer. The general equipment, materials, reagents, etc. used are all available from commercial channels unless otherwise specified.

[0034] See also Figure 1, which is a flow chart of the reversible skin phototransparency method using a living skin phototransparency reagent in the present invention. Specifically, the method includes the following steps: S1, depilating the skin of the subject, and obtaining skin in an original turbid state after washing and drying; S2, dripping a first solution onto the skin in an original turbid state, applying it evenly, performing a first reaction, and then removing the first solution to obtain pretreated skin; S3, applying a hydrogel containing edible pigments onto the pretreated skin, applying it evenly, and performing a second reaction to obtain phototransparent skin; S4, removing the hydrogel containing edible pigments on the phototransparent skin, and then scrubbing the skin to renature the skin tissue and restore it to the original turbid state.

[0035] Example 1 A living skin phototransparent reagent, comprising a first solution and a hydrogel containing acid red; wherein, in terms of volume percentage, the first solution comprises 50% sucrose, 5% thiazide and 45% polyethylene glycol; the hydrogel containing acid red is prepared by dissolving low melting point agarose in water and then adding acid red, wherein, in terms of mass percentage, the content of low melting point agarose is 0.2%, the content of water is 79.8%, and the content of acid red is 20%.

[0036] The method for reversible skin phototransparency using the living skin phototransparency reagent comprises the following steps: S1. After anesthetizing a two-month-old Balb / c mouse, the mouse's abdominal skin was depilated with a depilatory cream, and the depilatory cream on the skin surface was cleaned with 75% alcohol. Then, the skin surface was cleaned with saline, and the skin surface was blown dry with compressed air to obtain skin in its original turbid state; S2, adding the first solution to the skin in the original turbid state obtained in step S1, and gently massaging, and wiping with a cotton ball after reacting for 8 minutes to obtain pretreated skin; S3, applying the hydrogel containing acid red to the pretreated skin obtained in step S2, and gently massaging, and reacting for 10 minutes to obtain light-transparent skin; S4, removing the hydrogel containing acid red on the light-transparent skin obtained in step S3, and then scrubbing the skin with physiological saline to renature the skin tissue and restore it to its original turbid state.

[0037] The skin in the original turbid state obtained in step S1 of this embodiment, the light-transparent skin obtained in step S3, and the renatured skin tissue obtained in step S4 were respectively placed under an imaging device for observation. The results are as follows: Figure 2 shown.

[0038] from Figure 2It can be seen that after the abdomen of the mouse is transparentized by the living skin light-transparent reagent of the present invention, the abdominal tissue can become transparent. When the living skin light-transparent reagent is removed, the abdominal tissue returns to its original turbid state. The results show that the living skin light-transparent reagent of the present invention can make the abdominal tissue of the mouse light-transparent in a short time.

[0039] Example 2 A living skin phototransparent reagent, comprising a first solution and a hydrogel containing acid red; wherein, in terms of volume percentage, the first solution comprises 50% fructose, 2.5% thiazide and 47.5% polyethylene glycol; the hydrogel containing acid red is prepared by dissolving low melting point agarose in water and then adding acid red, wherein in terms of mass percentage, the content of low melting point agarose is 0.5%, the content of water is 74.5%, and the content of acid red is 25%.

[0040] The method for reversible skin phototransparency using the living skin phototransparency reagent comprises the following steps: S1. After anesthetizing two-month-old Balb / c mice, depilate the back skin of the mice with a depilatory cream, clean the depilatory cream on the skin surface with 75% alcohol, clean the skin surface with physiological saline, and blow dry the skin surface with compressed air to obtain skin in the original turbid state; S2, adding the first solution to the skin in the original turbid state obtained in step S1, and gently massaging, and wiping with a cotton ball after reacting for 5 minutes to obtain pretreated skin; S3, applying the hydrogel containing acid red to the pretreated skin obtained in step S2, and gently massaging, and reacting for 10 minutes to obtain light-transparent skin; S4, removing the hydrogel containing acid red on the light-transparent skin obtained in step S3, and then scrubbing the skin with physiological saline to renature the skin tissue and restore it to its original turbid state.

[0041] The skin in the original turbid state obtained in step S1 of this embodiment and the light-transparent skin obtained in step S3 were placed under an imaging device for observation respectively. The results are as follows: Figure 3 shown.

[0042] from Figure 3 It can be seen that after the living skin light-clearing reagent of the present invention is used to perform a transparent treatment on the mouse back, the back tissue can become transparent; the results show that the living skin light-clearing reagent of the present invention can make the mouse back tissue light-clear in a short time.

[0043] Example 3 A living skin phototransparent reagent, comprising a first solution and a hydrogel containing acid yellow; wherein, in terms of volume percentage, the first solution comprises 50% fructose, 2.5% thiazide and 47.5% polyethylene glycol; the hydrogel containing acid yellow is prepared by dissolving sodium hyaluronate in water and then adding acid yellow, wherein, in terms of mass percentage, the content of sodium hyaluronate is 0.5%, the content of water is 69.5%, and the content of acid yellow is 30%.

[0044] The method for reversible skin phototransparency using the living skin phototransparency reagent comprises the following steps: S1. After anesthetizing two-month-old Balb / c mice, the mouse legs were depilated with depilatory cream, and the depilatory cream on the skin surface was cleaned with 75% alcohol. Then, the skin surface was cleaned with normal saline, and the skin surface was blown dry with compressed air to obtain skin in the original turbid state; S2, adding the first solution to the skin in the original turbid state obtained in step S1, and gently massaging, and wiping with a cotton ball after reacting for 5 minutes to obtain pretreated skin; S3, applying the hydrogel containing acid yellow to the pretreated skin obtained in step S2, and gently massaging, and reacting for 10 minutes to obtain light-transparent skin; S4, removing the acid yellow hydrogel on the light-transparent skin obtained in step S3, and then scrubbing the skin with physiological saline to renature the skin tissue and restore it to its original turbid state.

[0045] The skin in the original turbid state obtained in step S1 of this embodiment and the light-transparent skin obtained in step S3 were placed under an imaging device for observation respectively. The results are as follows: Figure 4 shown.

[0046] from Figure 4 It can be seen that after the living skin light-transparent reagent of the present invention is used to perform transparency treatment on the mouse legs, the leg tissue can become transparent; the results show that the living skin light-transparent reagent of the present invention can make the mouse leg tissue light-transparent in a short time.

[0047] Example 4 A living skin phototransparent reagent, comprising a first solution and a hydrogel containing sunset yellow; wherein, in terms of volume percentage, the first solution comprises 50% sucrose, 5% thiazone and 45% polyethylene glycol; the hydrogel containing sunset yellow is prepared by dissolving sodium polyacrylate in water and then adding sunset yellow, wherein in terms of mass percentage, the content of sodium polyacrylate is 0.2%, the content of water is 59.8%, and the content of sunset yellow is 40%.

[0048] The method for reversible skin phototransparency using the living skin phototransparency reagent comprises the following steps: S1. After anesthetizing two-month-old Balb / c mice, the scalp of the mice was depilated with a depilatory cream, and the depilatory cream on the skin surface was cleaned with 75% alcohol. Then, the skin surface was cleaned with saline, and the skin surface was blown dry with compressed air to obtain skin in the original turbid state; S2, adding the first solution to the skin in the original turbid state obtained in step S1, and gently massaging, and wiping with a cotton ball after reacting for 8 minutes to obtain pretreated skin; S3, applying the hydrogel containing sunset yellow to the pretreated skin obtained in step S2, and gently massaging, and reacting for 10 minutes to obtain light-transparent skin; S4, removing the hydrogel containing sunset yellow on the light-transparent skin obtained in step S3, and then scrubbing the skin with physiological saline to renature the skin tissue and restore it to its original turbid state.

[0049] The skin in the original turbid state obtained in step S1 of this embodiment and the light-transparent skin obtained in step S3 were placed under an imaging device for observation respectively. The results are as follows: Figure 5 shown.

[0050] from Figure 5 It can be seen that after the mouse scalp is transparentized by the living skin light-transparent reagent of the present invention, the scalp tissue can become transparent; the results show that the living skin light-transparent reagent of the present invention can make the mouse scalp tissue light-transparent in a short time.

[0051] Comparative Example 1 In this comparative example, the living skin photoclearing reagent and the reversible skin photoclearing method using the living skin photoclearing reagent are basically the same as those in Example 2, except that the living skin photoclearing reagent only includes the first solution and does not include the hydrogel containing acid red.

[0052] The skin in the original turbid state obtained in step S1 of this comparative example and the light-transparent skin obtained in step S3 were placed under an imaging device for observation respectively. The results are as follows: Figure 6 shown.

[0053] from Figure 6 It can be seen that after the live skin light-transparent reagent of the present invention is used to perform transparent treatment on the back of mice, the back tissue can become transparent, but the transparent effect is obviously worse than that in Example 2.

[0054] Comparative Example 2 In this comparative example, the living skin photoclearing reagent and the reversible skin photoclearing method using the living skin photoclearing reagent are basically the same as those in Example 2, except that the living skin photoclearing reagent only includes a hydrogel containing acid red, and does not include the first solution.

[0055] The skin in the original turbid state obtained in step S1 of this comparative example and the light-transparent skin obtained in step S3 were placed under an imaging device for observation respectively. The results are as follows: Figure 7 shown.

[0056] from Figure 7 It can be seen that after the live skin light-transparent reagent of the present invention is used to perform transparent treatment on the back of mice, the back tissue can become transparent, but the transparent effect is obviously worse than that in Example 2.

[0057] Based on the results in Comparative Example 1, Comparative Example 2 and Example 2, when the living skin photo-clearing agent includes only one of the first solution and the hydrogel containing acid red, the transparentizing effect is significantly reduced; the results further show that in the present invention, by using a living skin photo-clearing agent that includes both the first solution and the hydrogel containing acid red, the two work synergistically to significantly improve the transparentizing effect.

[0058] It should be noted that the above embodiments all belong to the same inventive concept, and the description of each embodiment has its own focus. For matters that are not described in detail in some embodiments, reference may be made to the description in other embodiments.

[0059] The above-mentioned embodiments only express the implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A living skin light transparent reagent, characterized in that: comprising a first solution and a hydrogel containing food coloring; Wherein, the first solution comprises 40-60% of a carbohydrate substance, 1-10% of a penetration enhancer and 35-55% of an alcohol substance, in terms of volume percentage; The hydrogel containing edible pigment is prepared by dissolving a polymer compound in water and then adding the edible pigment. In terms of mass percentage, the content of the polymer compound is 0.1-1%, the content of water is 49-89.9%, and the content of the edible pigment is 10-50%.

2. The living skin light-transparent reagent according to claim 1, characterized in that: In the first solution, the sugar substance is selected from at least one of monosaccharides and their derivatives or polysaccharides and their derivatives.

3. The living skin light-transparent reagent according to claim 2, characterized in that: The monosaccharide and its derivatives are selected from at least one of fructose, glucose, galactose or mannose; the polysaccharide and its derivatives are selected from at least one of sucrose, lactose or maltose.

4. The living skin light-transparent reagent according to claim 1, characterized in that: In the first solution, the penetration enhancer is selected from at least one of thiazone, azone or acetone.

5. The living skin light-transparent reagent according to claim 1, characterized in that: In the first solution, the alcohol substance is selected from at least one of polyethylene glycol, polypropylene glycol or glycerol.

6. The living skin light-transparent reagent according to claim 1, characterized in that: In the preparation process of the hydrogel containing edible pigments, the polymer compound is selected from at least one of low melting point agarose, polyacrylamide, sodium polyacrylate or sodium hyaluronate.

7. The living skin light-transparent reagent according to claim 1, characterized in that: In the preparation process of the hydrogel containing edible pigment, the edible pigment is selected from at least one of acid red, acid orange, acid brown, lemon yellow or sunset yellow.

8. A method for reversible skin phototransparency using the living skin phototransparency reagent according to any one of claims 1 to 7, characterized in that: The steps include: S1. Depilating the skin of the subject, washing and drying the skin to obtain the skin in the original turbid state; S2, adding a first solution to the skin in an original turbid state, applying it evenly, performing a first reaction, and then removing the first solution to obtain pretreated skin; S3, applying the hydrogel containing edible pigments to the pretreated skin, and after applying evenly, performing a second reaction to obtain light-transparent skin.

9. The method according to claim 8, characterized in that The method also includes the steps of removing the hydrogel containing the edible pigment on the light-transparentized skin and then scrubbing the skin to remodel the skin tissue and restore it to its original turbid state.

10. The method according to claim 8, characterized in that In step S2, the first reaction time is 5-10 minutes; in step S3, the second reaction time is 8-15 minutes.

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