Multiparametric ultrasonographic process and analysis for validating pharmaceuticals, cosmetics, and biophysical effects

Abstract

The present invention patent application pertains to a multiparametric ultrasonographic process and analysis for validating pharmaceuticals, cosmetics, and biophysical effects, relating to an ultrasound analysis technique that uses three tools, along with their extensions and sub-tools, to assess changes in the tissue layers of an area under examination, describing and comparing the findings before and after the application of a cosmetic or pharmaceutical and evidencing their effects on the examined tissues, characterised in that multiparametric analysis is performed, combining high-resolution ultrasonography, high-power Doppler for microcirculation analysis, and strain and shear wave elastography, so as to carry out real-time, in vivo assessment of the effects of pharmaceuticals or cosmetics on the skin or scalp, painlessly and without the use of ionising radiation or any deleterious biological processes.

Description

[0001] "Multiparametric Ultrasound Process and Analysis for Validation of Drugs, Cosmetics, and Biophysical Effects"

[0002] INTRODUCTION

[0003] 001 This descriptive report of a Patent of Invention concerns a Multiparametric Ultrasonographic Process and Analysis for Validation of Drugs, Cosmetics and Biophysical Effects, hereinafter referred to as the Multiparametric Ultrasonographic Process and Analysis, which aims to evaluate "in vivo" and in real time, the reactions of the tissue layers of the skin and scalp, in a distinct and separate manner, an ultrasound analysis technique that uses three tools, with their unfoldings and sub-tools, to evaluate the changes found in the tissue layers of the area under study, describing and comparing the findings before and after application of a cosmetic or drug, highlighting its effects on the examined tissues.

[0004] 002 In this way, high-resolution ultrasound is used, specifically Doppler ultrasound of microcirculation / tissue perfusion, and of the stiffness or elasticity of each tissue layer, through Strain and Shear Wave Elastography, in a comparative way before and after, and can even be performed in several stages, after the application of drugs and cosmetics, being able to identify the biological reactions after the penetration of each active ingredient, and as a result we can highlight the obtaining of real-time images, being a painless, non-invasive method that does not present radiation or other risks to the patient's health.

[0005] FIELD OF APPLICATION

[0006] 003 The field of application of this Multiparametric Ultrasound Process and Analysis is intended for the Pharmaceutical Industry for incorporation into the research arsenal of the Pharmaceutical Industry, specifically in the development of drugs and cosmetics intended for skin and scalp. PURPOSE

[0007] 004 The purpose of this Multiparametric Ultrasound Processing and Analysis is to research and evaluate "in vivo" and in real time, the reactions of the tissue layers of the skin and scalp, serving the pharmaceutical industry when developing drugs and cosmetics for the skin and scalp and, in the case of consumers, avoiding problems when using these products.

[0008] STATE OF THE ART

[0009] 005 As is generally known among technicians and professionals in the field, cosmetic formulations are the basis for the development of a wide variety of beauty and skin and hair care products.

[0010] 006 () The development of any product generally encompasses formulation and laboratory testing before the product is placed on the market.

[0011] 007 According to the eBook "Alphabet of Cosmetic Formulations" from Farma Junior of the University of São Paulo, USP, the formulation of a cosmetic product encompasses several aspects represented in percentage and method of manufacture of the product, in addition, of course, to information on each ingredient, specifying its physical and chemical characteristics, functions, restrictions and possible interactions to be avoided.

[0012] 008 Each formulation is unique and manufactured according to the client's specifications and preferences in order to achieve the desired product result. The choice of each ingredient is based on its intrinsic physical-chemical characteristics and, when combined, these factors determine the texture, appearance, functionality, and other properties of the final product.

[0013] 009 Cosmetic formulations generally comprise:

[0014] Cosmetic actives, which are substances with a specific action to satisfy a particular need, such as moisturizing, anti-aging, and soothing effects. These are the components with the greatest commercial appeal.

[0015] . Adjuvants: These are capable of modifying the properties of the formula, consisting of emollients, humectants, surfactants, and thickeners. They can also have a corrective action, such as colorants, fragrances, preservatives, and antioxidants.

[0016] Vehicles and excipients: these are inert adjuvants that aim to give shape to the cosmetic, solubilize and carry the raw materials and active ingredients, that is, to take them to the site of action.

[0017] 010 The main components of a formulation for the aforementioned purposes are: Consistency Agents (or Thickeners); Moisturizing Agents; Antioxidants; Preservatives; Emollients; Emulsifiers; Essential Oils; pH regulator; Surfactants and Humectants.

[0018] 011 The functions of these components are described below:

[0019] - Consistency Agents

[0020] 012 Consistency agents or thickeners are substances capable of increasing the viscosity of cosmetic formulations, improving the rheological properties of stability, fluidity, feel (sensory) and appearance, ensuring better product performance. Most consistency agents can retain water in the skin and thus can act as moisturizing agents. Some also have emulsifying properties (ability to mix immiscible liquid phases, forming an emulsion) or gelling properties. Among the most used are Quaternized Guar Gum, Xanthan Gum and Glyceryl Monostearate.

[0021] - Moisturizing Agents

[0022] 013 Moisturizing agents have the property of permeating the stratum corneum of the skin and binding to the water molecules present there. Because of this property, the moisturizing agent is able to retain water molecules in the skin, preventing water from evaporating into the environment. Urea and glycerin are notable examples.

[0023] Antioxidants

[0024] 014 Antioxidants are responsible for interrupting, preventing, or delaying damage to cells caused by chain reactions of free radicals, unstable residual molecules that are highly reactive and produced by the body as a reaction to environmental pressure. Antioxidants provide protection against oxidative stress, which causes photoaging, wrinkling, degeneration of elastic tissue (elastosis), dryness, and skin pigmentation. Vitamin C, Vitamin E, and Ferulic Acid are prominent examples.

[0025] PRESERVATIVES;

[0026] 015 Preservatives are substances used to extend the shelf life of cosmetics because they inhibit the growth of microorganisms (such as bacteria, fungi, and yeasts) and thus prevent product deterioration, as well as ensuring their preservation during their expiration date and the safety and health of the consumer. Since there is no ideal preservative, the use of combinations or blends of preservatives is a very common practice because they are more likely to exhibit broad-spectrum activity. Furthermore, it is recommended that the preservative be present in a low concentration in the product to prevent the possibility of irritation and sensitization. Caprylyl Glycol stands out as the most commonly used.

[0027] - Emollients

[0028] 016 When used on the skin, the function of an emollient is to soften and smooth it, as it increases the skin's ability to retain moisture. Function: There is a wide variety of compounds that are emollients, and they generally have good spreadability. Jojoba oil and rosehip oil stand out in this regard.

[0029] - EMULSIFIER

[0030] 0.17 Promotes the mixing of immiscible phases, such as water and oil, reducing interfacial tension. Function: Emulsifiers are used to provide viscosity to creams and lotions. Notable examples include the use of Glyceryl Monostearate, Glyceryl Oleate, and Stearic Acid.

[0031] Essential Oils

[0032] Essential oils are natural active ingredients extracted from aromatic plants and can have various functions. They contain the "essence" of the plant's aromatic properties, that is, the volatile compounds (which quickly change from their solid or liquid state to a gas). In formulations, essential oils are mainly used for aromatization and to enhance effects. They are a good substitute for synthetic fragrances. Their function is due to their naturally intense odor and they can intensify the effects of cosmetics because of their properties that cause various effects on human skin. Because they are a hyper-concentrated form of the plant's active ingredient, essential oils should not be applied directly to the skin as they can cause irritation and allergic reactions. Furthermore, oils can be used individually or in complex combinations (blends). Among the most commonly used are Lavender Essential Oil, Rosemary Essential Oil, and Tea Tree Essential Oil.

[0033] pH regulator

[0034] 019 Its function is to adjust the pH of the formulation so that it has the correct pH for its respective purpose according to the pharmaceutical form of the product. The use of Citric Acid and Lactic Acid is highlighted.

[0035] - SURFACTANTS

[0036] 020 These are substances that have an affinity for both oils and fats as well as water, thus acting as a "combiner" of these immiscible phases.

[0037] 021 Surfactants can be classified according to their charge; therefore, we will cite an example for each classification:

[0038] Anionic: Sodium lauroyl sarcosinate. It is a biodegradable surfactant produced from the amino acid sarcozin. Being an anionic surfactant, it is incompatible with cationic surfactants. It can be used in personal care products, baby products, intimate hygiene products, and facial cleansing foams, since, in addition to its surfactant properties, it is also a foaming and conditioning agent.

[0039] Cationic: Ethioprine trimethyl ammonium chloride. This compound is a quaternary ammonium compound that can be used in hair care products because, in small quantities, it neutralizes the negative charges of the hair strand, providing softness. In addition to being a surfactant, it is also an emulsifier and antimicrobial.

[0040] Non-ionic: Decii glycoside. This compound is mildly aggressive and is made from sugar and coconut, making it suitable for sensitive skin, such as baby skin. In addition to being a non-ionic surfactant, it also acts as a cleansing agent and emulsion stabilizer. Because it is a non-ionic surfactant, it is compatible with other classes of surfactants.

[0041] Non-ionic: Cocamidopropyl Betaine. This compound is a co-surfactant, compatible with anionic, cationic, and non-ionic surfactants. It can be used in shampoos, bath products, and cleansing creams. This is because, in addition to being a co-surfactant, it is also a conditioning, thickening, and foaming agent.

[0042] - HUMECTANTS

[0043] 022 Humectants are hygroscopic components, meaning they have the property of absorbing moisture from the environment, moisturizing the stratum corneum (the outermost layer and surface of the skin). The absorption rate varies according to each substance. They contribute to skin hydration by forming a moisturizing film on it. Humectants form hydrogen bonds with water molecules, attracting them to the surface of the stratum corneum and retaining the skin's natural hydration, resulting in a less scaly appearance and a lower probability of cracking and irritation. Hyaluronic acid, glycerin, lactic acid, and sorbitol are prominent examples. 023 Another important item to consider refers to dermocosmetics, which are products with pharmacological active ingredients in their composition that act on the beauty and health of the skin.These products are able to penetrate deeper layers of the skin and, due to their active ingredients, can be used in various treatments, treating the skin from the inside out. Their action is more profound than that of common cosmetics, since these treat the skin only temporarily.

[0044] 024 The technology and compounds used in dermocosmetics, such as acids, amino acids, and proteins, act on the deeper layers of the dermis. Thus, they reduce or eliminate skin problems by treating them from their origin, with medium- and long-term results. They can also be used for prevention, delaying skin aging, dryness, and the appearance of blemishes, for example.

[0045] PROBLEMS TO SOLVE

[0046] 025 As is common knowledge among professionals in the field, both cosmetics and dermocosmetics, according to the regulations of the National Health Surveillance Agency, ANVISA, must meet various standards that attest to the quality of the product, which is why they are subjected to various laboratory tests and trials.

[0047] 026 Among the microbiological tests to which cosmetics are subjected, the following tests stand out:

[0048] Microbiological Tests

[0049] 027 Microbiological tests are fundamental in the analysis of cosmetics and pharmaceuticals, ensuring the safety and quality of products intended for consumers. These tests evaluate the presence, quantity, and type of microorganisms, such as bacteria, fungi, and yeasts, that can contaminate products during their manufacture, storage, or use.

[0050] Chemical and Physical Tests

[0051] 1)28 Chemical and physical tests are vital for determining the composition, quality, and stability of cosmetics and pharmaceuticals. These tests evaluate a range of properties and characteristics that directly influence the efficacy, safety, and acceptability of the products.

[0052] Shelf-life or shelf-time

[0053] 029 This test is used to determine the shelf life of cosmetic products, in other words, these studies are used to determine the expiration date of cosmetic products.

[0054] Challenge Test

[0055] 030 This test aims to determine the length of time a product maintains its functionality and safety under specified storage conditions. This allows companies to properly determine and label the expiration date of their products, ensuring that consumers receive effective and safe products throughout the recommended usage period.

[0056] 031 With regard to dermocosmetics, the tests for approval and release by the Health Surveillance Agency are more rigorous and actually prove and demonstrate their safety and efficacy.

[0057] 032 According to current legislation, the three most common tests for dermocosmetics are:

[0058] Efficacy test: the result is the perception of those who used the product, who give their opinion on how much their skin improved.

[0059] Clinical test: the result is determined by the perception or evaluation of a dermatologist.

[0060] Dermatological test: verifies that the product is safe for use, according to an evaluation by a dermatologist.

[0061] 033 It is worth highlighting that the only test performed to verify penetration is the test called In Vitro Cutaneous Absorption, where Cutaneous Absorption is understood as the passage of the active ingredient(s) through the epidermis, reaching the viable epidermis (formed by cells with intense proliferative capacity) or the dermis. 034 From the analysis of the State of the Art, including the part corresponding to the Tests, it is concluded that the Multiparametric Ultrasonography Process presented here is not performed on any type of product currently manufactured and marketed, either in Brazil or in the rest of the world.

[0062] PATENTIAL BACKGROUND

[0063] 035 With a view to placing a Multiparametric Ultrasonographic Process and Analysis on the market, prior art searches were carried out in the INPI Database and no identical or similar documents were found.

[0064] TECHNOLOGICAL ADVANCEMENT

[0065] 036 As it develops, the technological advancement presented by the Multiparametric Ultrasound Process and Analysis refers to the examination of cosmetic products by multiparametric analysis, which combines high-resolution ultrasound, high-power Doppler for microcirculation analysis, and strain and shear wave elastography, allowing the evaluation of the effect of drugs or cosmetics on the skin or scalp in real time and "in vivo", painlessly, without the use of ionizing radiation or any deleterious biological process.

[0066] ADVANTAGES

[0067] 037 The advantages relate to real-time and “in vivo” analysis, painlessly, without the use of ionizing radiation or any other process, of the reactions of the tissue layers of the skin and scalp after the use of cosmetic products.

[0068] DESCRIPTION OF THE METHODOLOGY

[0069] 038 To obtain a complete and thorough understanding of how the Multiparametric Ultrasound Process and Analysis is structured, it is important to highlight that it corresponds to an ultrasound analysis technique that uses three tools, with their sub-tools and components, to evaluate the changes found in the tissue layers of the area under study in this specific case, describing and comparing the findings before and after the application of a cosmetic or drug - highlighting its effects on the examined tissues.

[0070] 039 Ultrasound offers several advantages, most notably obtaining images in real time, being a painless, non-invasive method that does not involve radiation or other health risks to the patient.

[0071] 040 B-mode, known as Bright Mode, consists of the actual ultrasound, in grayscale, used for the anatomical investigation of organic tissues in real time. These images aim to study the morphology of the tissue layers that make up the skin and scalp, from the epidermis, superficial dermis (or papillary dermis), deep dermis (or reticular dermis), adipose layer (or hypodermis) and fascia or aponeurosis depending on the region studied, whether it is the face, scalp or other body regions.

[0072] 041 Doppler ultrasound reveals changes in blood flow and tissue perfusion, as well as assessing peak systolic pressure and blood flow resistance index in the tissues studied at their analysis sites.

[0073] 042 Evaluation of the spatial distribution of blood circulation and microcirculation, the quantity and quality of blood vessels supplying the skin or scalp with its follicles, as well as the caliber of such vessels, by amplitude Doppler flowmetry analysis. Demonstrating the greater capillary vascular capacity for blood irrigation of the investigated territory.

[0074] 043 Arterial spectral assessment, of the maximum amplitude of the peak systolic velocity and the resistance index, which is the ratio between systole and diastole (derived from heartbeats), by color Doppler velocimetry. Analysis of regional venous return can also be performed, to determine if it is increased, stable, or reduced.

[0075] 044 In turn, ultrasound elastography is a modality that aims to evaluate the stiffness or elasticity of a region, in order to better characterize pathologies that alter the fibroelastic characteristics of a given area.

[0076] 045 Qualitative Electrography, by compression or Strain Wave, evaluates the greater stiffness or greater elasticity of tissue after mechanical compression stimuli performed with the transducer itself in an area of ​​interest.

[0077] 046 This technique allows for color-map characterization according to the viscoelastic properties of each fabric, with variations ranging from "soft" (highest elasticity), intermediate, and "hard" (highest stiffness). In this method, we do not have numerical values ​​to define stiffness, but we can perform comparative studies of stiffness in neighboring fabrics and obtain numerical relationships between them (example: Point A is 3 times stiffer than point B).

[0078] 047 Quantitative Elastography, by shear or

[0079] Shear Wave. A technique that allows characterization in numerical values ​​in kPa or m / s (kilopascals or meters per second).

[0080] 048 DESCRIPTIVE ANATOMY OF THE AREAS FOR

[0081] APPLICATION OF THE TECHNIQUE

[0082] 049 The figures above demonstrate the layers of the skin and scalp, which consists of a differentiated area of ​​skin, where the thickness of a healthy scalp will vary according to the age, sex and biotype of the patient, being in children, depending on age, between 2-3 mm and in adults 5-7 mm, highlighting that healthy skin varies according to the region of the body (e.g.

[0083] 050 The skin on the back is thicker, the skin on the forearm is thinner), according to ethnicity and age.

[0084] 051 Here are some illustrative examples:

[0085] - Research on the active ingredient used in shampoos containing Officinol, aiming to evaluate the ultrasonographic changes found in the scalp before and after application of shampoos containing the active ingredient OFFICINOL (Sugar cane) in different concentrations, demonstrating the permeation of the product through multiple parameters (B-mode or high-resolution ultrasound, high-power Doppler showing vascular map and spectral Doppler, and elastography (Shear Wave and / or Strain Wave), evaluating the stiffness / elasticity of the area of ​​interest).

[0086] 052 This active ingredient is multifunctional, but primarily a strong antioxidant. It has already been tested for rejuvenation and improvement of skin tone, and more recently, in vitro research has been conducted with it for stimulating hair growth.

[0087] 053 The methodology applied corresponds to:

[0088] - Initially, a preliminary analysis of the volunteers' scalp is performed, acquiring images in B-mode, Doppler with perfusion mapping, spectral analysis, and elastography of the area of ​​interest.

[0089] 054 Next, the shampoo is applied to the volunteers' scalp along the midline, with a light local massage, the product is left on for 3 minutes and then rinsed thoroughly.

[0090] 055 Acquisition of new images through multiparametric analysis starting 15 minutes after application, highlighting the changes found in comparison with the Results:

[0091] - The study was conducted with the objective of demonstrating en vivo and in real time the permeation capacity of the studied active ingredient through a liquid vehicle, in the form of shampoo - at concentrations of 1% and 2.5%, evidenced by the changes found in the images obtained by the multiparametric ultrasound study as illustrated in the following images:

[0092] - Multiparametric Ultrasound Study of the Scalp In Vivo to Evaluate the Permeation of the Active Ingredient Officinol after application in a shampoo formula.

[0093] 056 Hair loss represents the most recurrent and stressful clinical complaint faced by dermatologists. Androgenetic alopecia (AGA), alopecia areata (AA), and telogen effluvium (TE) are the most common types of hair loss, with AGA being the most prevalent, affecting almost 70% of men and 40% of women worldwide.

[0094] 057 The etiology and pathogenesis of AGA remain undefined; however, accumulating evidence has indicated that increased levels of dihydrotestosterone (DHT), overexpression of the androgen receptor gene, miniaturization of hair follicles in androgen-sensitive areas, genetic predisposition, and emotional stress contribute to its development.

[0095] 058 There are studies that indicate that administering medication through the scalp offers an available delivery method for drugs, particularly hydrophilic drugs. These studies were mostly conducted through histological analyses, using tissue biopsies.

[0096] 059 The in vitro test performed with Offieinol demonstrated efficacy in preventing premature aging of follicles, at concentrations of 1% and 2.5%.

[0097] In this study, we demonstrate the changes found in vivo in the scalp using a non-invasive tool, without the use of radiation or contrast, making it possible to compare the data obtained at time intervals, depending on the type of analysis desired.

[0098]

[0099]

[0100]

[0101] This 2.5% shampoo formula was applied to a 48-year-old female volunteer with early-stage androgenetic alopecia.

[0102]

[0103] Doppler

[0104] Perfusion map:

[0105]

[0106] Spectral Doppler

[0107]

[0108] 061 In conclusion, based on the results obtained, we can affirm that the active ingredient permeated the scalp after local application, according to the described methodology. This effect demonstrates the ability of the described active ingredients to be delivered through the scalp, showing modification of the tissue layers immediately after application, indicating absorption of the studied substances by this method of use. It is important to highlight the greater increase in vascularization at the 1% concentration of OFFICINDL Shampoo, however, with an increase in tissue rigidity.

[0109] 062 At a concentration of 2.5% of the aforementioned product, we observed a smaller increase in the distribution of the vascular network. In all spectral Doppler analyses, there was an increase in peak systolic velocities with a reduction in the Resistance index (ratio between systolic velocity and diastolic velocity), and in elastographic analyses, we evidenced an increase in tissue stiffness at the 1% concentration in all layers, and, conversely, we observed a significant increase in elasticity in the tissue layers at the 2.5% concentration.

[0110] 063- Multiparametric Ultrasound Study of the Scalp In Vivo for Evaluation of the Penneation of “ACTIVE INGREDIENT BLEND”

[0111] 064 Pre- and post-application analyses of drugs on the scalp and face, showing by multiparametric ultrasound the immediate changes resulting from the infusion of said substances, performed by electroporation infusion equipment with different pulse frequencies.

[0112] 065 Comparative evaluation before and 5 minutes after infusion of an "active ingredient blend" with equipment that promotes infusion by electroporation, without the use of needles, in a 52-year-old female patient with Androgenetic Alopecia by multiparametric Ultrasound (High-resolution B-mode, Ultrasensitive Doppler and Shear Wave Elastography) in equipment that uses 14 to 33 MHz transducers.

[0113] 066 Infused area:

[0114] Scalp - Midline

[0115] 067 Demonstrating tissue layers of the epidermis, superficial dermis, deep dermis, and hypodermis with good differentiation of their boundaries. Note the thinning, however, maintaining the regularity of the contours in the deep dermis.

[0116] 068 Mode B (Bright Mode)

[0117] EP - epidermis;

[0118] DS - superficial dermis;

[0119] DP - deep dermis;

[0120] HYPOD - hypodermis

[0121] 069 Doppler - Perfusion Map:

[0122] -Distribution of the vascular map in the tissue layers, showing a predominance in the deep dermis.

[0123] 070 Spectral Doppler

[0124] - Good peak systolic velocity with an intermediate resistance index (RI) in arteriolar analysis at the limits of the deep dermis.

[0125] - Systolic peak: 8.4 cm / s

[0126] -IR: 0.64

[0127] 071 Elastography Shear Wave:

[0128] Elastogram showing a very similar degree of stiffness between the layers of the scalp, being:

[0129] -Superficial dermis: 3.86 m / s

[0130] -Deep dermis: 3.93 m / s

[0131] -Hypodermis: 3.83 m / s

[0132] -Mode 3 (Bright Mode)

[0133] Scalp - 5' post-infusion

[0134] -Increased echogenicity of the superficial dermis, making it difficult to distinguish between the superficial and deep dermis.

[0135] 072 Doppler - perfusion map

[0136] - Diffuse increase in vascular perfusion of the superficial and deep dermis and an increase in the superficial hypodermis region.

[0137]

[0138] 073 Spectral Doppler

[0139] - Arteriolar spectral analysis at the limits of the deep dermis demonstrating a reduction in peak systolic velocities and a slight increase in the resistance index (RI).

[0140] - Peak systolic rate: 2.8 cm / s

[0141] - IR: 0.67

[0142] 074 Elastography Shear Wave

[0143] -Elastogram demonstrating an increase in elasticity in the superficial dermis and hypodermis of approximately 4% and an increase in stiffness in the deep dermis of 6%.

[0144] - Superficial dermis: 3.70 m / s

[0145] - Deep dermis: 4.16 m / s

[0146] - Hspoderme: 3.68 m / s

[0147] 3 - FACE - INFUSION OF HALURONIC ACID IN THE RIGHT ZYGOMATIC ARCH BY ELECTROPORATION, AT A RATE OF 5 PULSES PER SECOND.

[0148] 075 Pre-infusion

[0149] - Comparative evaluation before and 5 minutes after hyaluronic acid infusion with Mesoject Gun in a 54-year-old female patient, using multiparametric ultrasound (high-resolution B-mode, ultrasensitive Doppler and Shear Wave Elastography) on equipment that uses .14 to 33 MHz transducers.

[0150] - Infused area:

[0151] -RIGHT ZYGOMATIC ARCH

[0152] 5 pulses / second

[0153] Mode B

[0154] -Face ~ zygomatic arch D - pre-infusion

[0155] - Demonstrating the lesional layers of the epidermis, superficial dermis, deep dermis, and hypodermis with good differentiation of their boundaries. 076 Doppler - perfusion map

[0156] -Face - zygomatic arch D - pre-infusion

[0157] -Perifusion map demonstrating vascular network with typical distribution.

[0158] 077 Spectral Doppler

[0159] - Face - zygomatic arch D - pre-infusion

[0160] - Usual speed, with a moderate resistance index in an arteriole located in the deep dermis.

[0161] -Systolic peak: 6.0 cm / s

[0162] - IR: 0.67

[0163] 078 Elastography Shear wave

[0164] - Face ~ zygomatic arch D - pre-infusion

[0165] Elastograin showing a very similar degree of stiffness between the dermal layers of the face, being:

[0166] - Superficial dermis: 1.40 m / s

[0167] - Deep dermis: 0.1,37 m / s

[0168] Hypodermis: 1.41 m / s

[0169]

[0170] Mode B

[0171] - Face - xygomatic arch D - 5' post-infusion

[0172] - Mode B demonstrating tissue layers of the epidermis, superficial dermis, deep dermis, and hypodermis with slight blurring of their boundaries.

[0173] 079 Doppler perfusion

[0174] - Face - zygomatic arch D - 5' post-infusion

[0175] - Demonstrating a diffuse increase in the vascular network of the superficial and deep dermis.

[0176] 080 Spectral Doppler - Face ~ zygomatic arch D - 5' post-infusion

[0177] - Significant increase in peak systolic velocity and reduction in the resistance index in arterioles at the limit of the deep dermis.

[0178] - Peak systole: 15.2 cm / s

[0179] - IR: 0.61

[0180] 081 Elastography Shear Wave

[0181] - Face - zygomatic arch D - 5' post-infusion

[0182] - Elastogram showing increased stiffness comparatively in all tissue layers, being approximately 8.5% in the DS, 7.5% in the DP and 6% in the hypodermis.

[0183] - Superficial dermis: 1.53 m / s

[0184] Deep dermis: 1.48 m / s

[0185] Hypodermis: 1.50 m / s 4 - FACE - INFUSION OF HYALURONIC ACID IN THE LEFT ZYGOMATIC ARCH BY ELECTROPORATION, AT A RATE OF 3 PULSES PER SECOND.

[0186] 082 Comparative evaluation before and 5' after hyaluronic acid infusion with Mesoject Gun in a 54-year-old female patient, by monoparametric ultrasound (high-resolution B-mode, ultrasensitive Doppler and Shear Wave Elastography) using equipment with transducers from 1.4 to 33 MHz.

[0187] Mode B

[0188] - Face - zygomatic arch E - pre-infusion

[0189] - B-mode demonstrating tissue layers of the epidermis, superficial dermis, deep dermis, and hypodermis with good differentiation of their boundaries.

[0190] 083 EP-epidemism; DS- superficial dermis; DP- deep dermis; HIPOD- hypodermis

[0191] - Face - zygomatic arch E - pre-infusion

[0192] 084 Doppler - perfusion map

[0193] - Perfusion map demonstrating vascular pattern with usual distribution

[0194]

[0195] -Face - zygomatic arch E - pre-infection

[0196] 085 Spectral Doppler

[0197] -Usual speed, with a moderate resistance index in localized aryerfoía in the deep dermis.

[0198] - Peak systolic rate: 3.6 cm / s

[0199] ~ IR: 0.66

[0200] - Knife - zygomatic arch E - pre-infusion

[0201] 086 Elastography Shear wave

[0202] Elastogram showing very similar degrees of stiffness between the dermal layers of the face, being:

[0203] - Superficial dermis - 2.0 m / s

[0204] - Deep dermis - 1.86 m / s - Hypodermis - 1.80 m / s

[0205] - Face - zygomatic arch E - 5' post-infusion

[0206] Mode B

[0207] ~ Mode B demonstrating tissue layers of the epidermis: superficial dermis, deep dermis, and hypodermis with slight blurring of their boundaries.

[0208] - Face - zygomatic arch E ~ 5' post-infusion

[0209] 087 Perfusion Doppler

[0210] - Demonstrating a diffuse increase in vascular network, mainly in superficial dermatitis.

[0211] - Face - zygomatic arch E - 5' post-infection

[0212] 088 Spectral Doppler - Large increase in peak systolic velocity and slight reduction in resistance index in arteriole at the limit of the deep dermis.

[0213] - Peak systolic rate: 22.6 cm / s

[0214] -IR: 0.60

[0215] - Face - zygomatic arch E - 5' post-infusion

[0216] 089 Elastography Shear Wave

[0217] - Elastogram showing a reduction in stiffness compared to all tissue layers, being approximately 5% in the DS, 8.5% in the DP, and 2% in the hypodermis.

[0218] - Superficial dermis - 1.90 m / s

[0219] - Deep dermis - 1.77 m / s

[0220] Hypodermis - 1.77 m / s

[0221]

[0222] CONCLUSION

[0223] 090 It is verified by all that has been described and illustrated that this is a Multiparametric Ultrasonographic Process and Analysis for Validation of Drugs, Cosmetics and Biophysical Effects, which perfectly fits within the norms governing the Patent of Invention due to filling an important gap existing in the market.

Claims

CLAIMS 1) “MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS”, which corresponds to an ultrasound analysis technique that uses three tools, with their sub-tools and components, to evaluate the changes found in the tissue layers of the area under study, describing and comparing the findings before and after the application of a cosmetic or drug and highlighting its effects on the examined tissues, characterized by performing multiparametric analysis combining high-resolution ultrasound, high-power Doppler for microcirculation analysis and Strain and Shear Wave Elastography in order to evaluate the effect of drugs or cosmetics on the skin or scalp in real time and “in vivo”, painlessly, without the use of ionizing radiation or any deleterious biological process. 2) "MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS", according to claim 1, characterized by applying high-resolution ultrasound in B-mode, known as Bright Mode, for anatomical investigation of organic tissues in real time and analyzing the morphology of the tissue layers comprising the skin and scalp, from the epidermis, superficial dermis (or papillary dermis), deep dermis (or reticular dermis), adipose layer (or hypodermis) and fascia or aponeurosis depending on the region studied, whether it is the face, scalp or other body regions. 3) “MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS”, according to claim 1, characterized by applying high-power Doppler for the analysis of microcirculation and highlighting the changes found in blood flow and its tissue perforation, as well as evaluating the systolic peak and flow resistance index. Blood in the tissues studied at their analysis sites through the evaluation of spatial distribution and arterial spectral evaluation. 4) "MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS", according to claims 1 and 3, characterized by allowing, through amplitude flowmetric analysis, the evaluation of the spatial distribution of blood circulation and microcirculation, the quantity and quality of blood vessels nourishing the skin or scalp with its follicles, as well as the caliber of such vessels. 5) “MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS”, according to claims 1 and 3, characterized by allowing, through Doppler color velocimetry, the arterial spectral evaluation, the maximum amplitude of the peak systolic velocity and the resistance index, which is the ratio between systole and diastole resulting from heartbeats. 6) “MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS”, according to claim 1, characterized by the application of Strain Wave Elastography or Qualitative Elastography, to evaluate the greater stiffness or greater elasticity of tissue after mechanical compression stimuli performed with the transducer itself in an area of ​​interest. 7) “MULTIPARAMETRIC ULTRASONOGRAPHIC PROCESS AND ANALYSIS FOR VALIDATIONS OF DRUGS, COSMETICS AND BIOPHYSICAL EFFECTS”, according to claim 1, characterized by the application of Quantitative Elastography, by shear or Shear Wave to characterize elasticity in numerical values ​​in Kpa or m / s (Kilopascal or meters per second).

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