CA2378155A1

CA2378155A1 - Colloidal composition for esthetic correction

Info

Publication number: CA2378155A1
Application number: CA002378155A
Authority: CA
Inventors: Mateus Sommer Neto
Original assignee: Individual
Current assignee: Individual
Priority date: 1999-08-02
Filing date: 2000-08-01
Publication date: 2001-02-08
Also published as: MXPA02001210A; JP2003519093A; WO2001008632A2; WO2001008632A3; EP1200057A1; BR9904471A; AU6142500A

Abstract

The present invention refers to a method and a chemical composition of carboxy glyconate hydrolactic or magnesium, to be employed with the aim of correcting dermatological disorders. More specifically, the present invention refers to a composition to be applied, in humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the na-sogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight.

Description

COI~LOIDAI~ COMPOSITION FOR ESTHETIC CORRECTION
FIELD OF THE INVENTION
This invention refers to a method and a chemical composition to be employed with the aim of correcting dermatological disorders. More specifically, the present invention refers to a composition to be applied, in humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of trie sKln associated to natural aging, or through the action of external agents, such as sunlight.
BACFCGROUND OF THE INVENTION
Human skin, when intact, is a barrier for many natural and synthetic substances. Cosmetic and pharmaceutical agents can be pharmacologically effective through oral administration or other systematic administrations of these agents, however, many agents presently known are barely or totally ineffective for topical application to the skin.
Topical effectiveness of pharmaceutical agents depends on two major factors: (a) the biological availability of an active ingredient in the topical preparation, and, (b) the absorption, penetration and distribution of the active ingredient in the area of the skin to be treated.
The formation of wrinkles may be due to natural aging and/or damages caused by the sun. A majority of the fine wrinkles on the face are caused by natural or congenital aging, whilst the deeper wrinkles on the face are the consequence of a skin with acne or action of the sun. Despite the real mechanism of the formation of wrinkles remaining unknown, the formation of the fine visible wrinkles due to the reduction in the number and diameter of the elastic fibers in the papillary dermis is accepted, as well as the atrophy of the dermis and reduction of adipose tissue.
Histopathological and electronic microscopy studies ,indicate that the thicker wrinkles are caused due to an excessive deposit of abnormal elastic materials in the superior dermis and thickening of the skin.
Despite the fact that imperfections of the skin do not possess any physiological significance, these imperfections may IS cause serious physiological problems to the human being that possesses them. In many societies, the bearers oz tnese imperfections dislike showing the signs of aging, and wrinkles are the greatest preoccupation of the human race since time immemorial .
Implants and grafts presently available used to correct depressions and corporal imperfections demonstrate the difficulties in handling, complications and deficiency in long term results.
It is known in the state of the art the use of various vehicles with the aim of correcting esthetical imperfections.
One of these vehicles is bovine collagen, which is a long chain protein with a high risk of allergy and passive to transmit viruses, such as bovine encephalitis, or, also, lasting future complications, such as inflammatory processes, intense allergic phenomena and, mainly, autoimmune diseases (for instance, rheumatoid arthritis).
Another vehicle also known in the state of the art is liquid silicone, however liquid silicone produces siliconomas, and has not been entirely approved by the authorities for medical use.
Amongst the classic treatments for the elimination of wrinkles, esthetic surgery may be mentioned, which, whilst eliminating wrinkles, presents numerous disadvantages, including the scars left on the skin by these surgeries, considering that these scars are, frequently, very difficult to occult.
Another method for eliminating wrinkles includes the use of implants made of various materials. For instance, the Brazilian patent application PI 9503052-2 filed on 22 June 1995, in the name of Miguel Marques 0liveira Jr., incorporated herein for reference. The PI 9503052-2 describes an alloplastic surgical implant for increase and/or esthetic modeling for esthetic definition of the forehead. A prosthesis built in accordance with the PI 9503052-2, is made in semi-rigid solid silicone, and implanted in the frontal region for the correction of deficiencies, however, the disadvantages of this implant are numerous and include rejection, allergic reactions and the effectiveness is limited to a restricted area.

Brazilian patent PI 8907235-9, granted on the 24 September 1996 to Dr. Martin Lemperle, refers to compositions for use in esthetic medicine, and is incorporate herein for reference. The PI 8907235-9 describes a composition for filling imperfections at skin level. This composition employs polymethylmetacrilate microballoons in a suspension medium. The suspension medium consists of water, alcohols, or a mixture of these, and also a tensoactive, such as Tween 80. However, as this composition is not stable, it requires to be kept under rigorous storage condit;-ons and under constant refrigeration.
It is now being discovered that dermatolcgical imperfections may be corrected without presenting the disadvantages related ~.~ the state of the art.
SUMMARY OF THE INVENTION
The present invention refers to a method and a chemical composition, particularly to be employed with the aim of correcting dermatological disorders, however, that may be used for any other purpose in reconstructive surgery.
One objective of the present invention is to present a method and a composition possessing a perfect combination of materials, which are practical concerning storage, not requiring refrigeration, are easily handled by the doctor, do not induce allergies or infections, are not cancerigenous, are biocompatible and, finally, are low cost.
One of the main objectives of the present invention. is to offer a new method and composition to correct dermatological disorders, such as the elimination of wrinkles in the skin of humans, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation cf the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural 5 aging, or through the action of external agents, such as sunlight.
The composition of the present invention is applicable through the injection of the composition immediately below the area of the skin where the dermatologicai imperfections occur.
The composition of the present invention is not, substantially, absorbable by the human body, and, consequently, will not be rejected by the human being, and will also not cause allergic reactions.
In addition, the method of injecting the composition, in accordance with the present invention, is easily and dependably applicable, without requiring the anesthetization of the patient.
DETAILED DESCRIPTION OF THE INVENTION
The composition, in accordance with the present invention, is composed of a colloidal formulation termed carboxy glyconate hydrolactic of magnesium.
This formulation consists of:
(a) approximately 5 to 70g of an ether of cellulose;
(b) approximately 800 to 1300m1 of Ringuer lactate;
(c) approximately 2 to 20g of EDTA;
(d) 20 to 120m1 of calcium glyconate; and, (e) optionally, between 0 and loo xylocaine.

The quantities of the ingredients may vary within the optimal ranges until adequate viscosity is attained.
Amongst the polymers of ether of cellulose, those with low molecular weight or low viscosity are preferable, such as:
hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcelluiose, hydroxybutylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose of sodium, carboxymethylcellulose of calcium, methylcellulose and/cr ethylceliulose.
The Ringuer lactate is an inert solution composed of sodium chloride, potassium chloride, dehydrated calcium chloride, sodium lactate and water.
The EDTA, termed trihydrated ethylenediamine trisodium tetracetate is chemically inert, and consequently, appropriate for medical use, because apart from not decomposing in other substances, it does not react with other chemical bodies whether in the composition of the invention or in the human body. Its chemical formula is Cio-H~6-N?-Oe.3Na.
The preparation process of the composition of the present invention consists in mixing all the ingredients in stainless steel tubes, followed by agitation. The temperature of the mixing may vary between 30°C and 70°C, preferably at around 50°C, and for a time period for approximately 3 hours.

This procedure is repeated twice more, until obtaining the final product.
At this stage a colloidal solution is obtained.
The resulting colloidal product is then filtered, and its pH is verified so as to present a value of approximately 6Ø
The density of the product shall present a value equal to 1.
The resulting product, that is, the carboxy glyconate hydrolactic of magnesium composition, already having the pH and density required, -is injected into vials of 20m1, and after labeling, are ready for distribution.
To illustrate =he present invention, experiments in animals were performed. These experiments were based on the investigation of the reactions occurring in the issues of the animals after the application of the exogenous materials in accordance with the present invention.
EXPERIMENT I
Comparative histoloaical study of the cellular alterations and of the extracellular matrix resulting from the injectable implant of the carboxy alyconate hydrolactic of magnesium colloid in the hvpodermis of albino rats. Preliminary results In Experiment I the possible different tissue reactions resulting from the injection of carboxy glyconate hydrolactic of magnesium were evaluated in a group of 108 rats, obtaining samples in the 1st, 2na~ q~n~ 8rh and 16th weeks, with another 20 rats injected with a physiological solution of sodium chloride, kept in identical conditions to act as a control.

Materials and Method Seventy-two adult rats, albino, females of the Wistar breed, were superficially anesthetized through the inhalation of ether, trichotomized on the back (interscapular region) with an electric hair clippers, selecting, on the median dorsal line, an area where two surgical stitches were applied, placed sagitally, with black line (having a distance of l.5cm between them). On the back, in the interscapular region, a first group of 36 rats was injected with 0.5m1 of the carboxy glyconate hydrolactic of magnesium colloidal compositior_, and the rats were placed in three appropriate plastic cages (l3cm each) with a bottom of sterile wood shavings and supplying them wit:: food and filtered water ad libitum. A second group ,.~ 36 rats was, treated similarly, however a physiological solution of sodium chloride was injected and these rats were kept in identical conditions to act as a control.
After l, 2, 4 and 8 weeks, 14 animals (sever injected with carboxy glyconate hydrolactic of magnesium and seven inected with the physiological solution were put down in deep anesthesia with ether, and, then had fragments cf skin, kidney, liver, spleen, heart and lungs removed, at the same time that their principal lymph nodes were examined for eventual histopathological studies, in the case of growth being found.
In the 16th week, the remaining animals were put down.
The fragments were fixed in a solution of formaldehyde at 10 o in PBS, at 4°C, for a period of 3 to 5 days, after which they were washed for 15 minutes in running water. In addition, a second fragment of skin from the region injected was removed from each animal, and fixed for 4 hours in a solution of paraformaldehyde at 5o in PBS for later immuno-histochemical study and, after washing in PBS, the fragment was treated in the same way as the others for histological processing.
All the fragments were then dehydrated in solutions of increasing concentrations of ethanol, diaphanized by xylene, to be later impregnated and, finally, included in paraffin. Cuts of 5 micrometers were obtained using a Spencer 820 rotative microtome from American Optical Co., and, those previously fixed in formaldehyde-PBS, after being mounted on histological slides, were dyed by the Hematoxillin-Eosin technique. The preparation could then be examined by an Optiphot model Nikon microscope, coupled to an automatic AFX II photometer with a IS 35mm camera, thus obtaining photomicrographies on 100 ASA
Kodacolor film from Kodak.
During the first phase of observations, it is possible to observe in the first week, through the visualization of the skin from the animals injected with carboxy glyconate hydrolactic of magnesium, that droplets of colloid (identifiable as amorphous and basophilic material) are surrounded by mononuclear cells, which develop intense infiltration in the area of the injection around the annexes below the epidermis. In the second week, the infiltration persists and macrophages can already be observed around the droplets of colloid, as well as giant multinuclear cells. In the fourth and eighth weeks, macrophages appear, either molding incipient capsules or infiltrating between the colloid droplets. Finally, in the 16th week, macrophages, fibroblasts and conjunctive matrix surround conglomerates of colloid, still distinguishable by their basophilia.
5 In addition, the observation of images of the retraction of the blood plasma fixed inside the blood vessels close to the endothelial surface, which could be confused with a possible presence of minute microballoons in the blood, become, in reality, impossible to be mistaken, as in the blood vessels of 10 ~~e animals injected with NaCl, such ,images are also present.
No signs of neoplasia were detected in the fragments of the skin studied.
From the second week of experiment, the colloid is already retained in the granuioma, in stage of formation, present in a general manner below the muscle, but that can pass the muscular layer and reach the dermis. This granuloma evolves until constituting a sac containing the colloid inside still surrounded by the macrophages. In tt~.e 16'h week, conjunctive matrix can be observed between the macrophages and fibroblasts as part of the conjunctive capsule surrounding the granuloma.
Conclusion of Experiment I
From the observations of Experiment I, it can be concluded that the injection of carboxy glyconate hydrolactic of magnesium, in the hypodermis of albino rats, promotes the 35 formation of a granuloma reacting to a foreign body, localized preferentially at the place of injection, that is, in the inferior part of the skeletal muscular layer (present in the rat) of the hypodermic. In some situations, however, it reaches the supra muscular region of the hypodermic, attaining deep dermis. This granuloma becomes stable around the second week, after the injection, and the carboxy glyconate hydrolactic of magnesium colloid can be visualized in an apparently stable form surrounded by giant foreign body cells. After the eighth week, the granulomas becomes surrounded by conjunctive capsules that extend septa inwards, turning the granulomas lobulated.
The presence of neoplasia was not observed.
EXPERIMENT II
Comparative histoloaical study of the cellular alterations and the extracellular matrix resulting from the injected implant of the carboxv alvconate hydrolactic of magnesium colloid in the hvpodermis of albino rats. Evaluation of the matrix In Experiment II, adult female albino rats of the Wistar breed were used.
This Experiment II was an evaluation of tissue alterations resulting from the injected implant of carboxy glyconate hydrolactic of magnesium in 108 albino rats, injecting these 108 rats with carboxy glyconate hydroiactic of magnesium in the previously trichotomized dorsal interscapular region. Another 20 rats were injected with a physiological solution of sodium chloride, and were kept in identical conditions to act as a control.
Material and Methodolo The methodology used for obtaining the tissue of the animals is described below.

The albino rats, after being put down in deep anesthesia, had fragments of skin, kidney, liver, spleen, heart and lung removed. These fragments were fixed in a solution of formaldehyde at 10% in PBS, at 4°C, for a period of 3 to 5 days, after which they were washed for 15 minutes in running water. All the fragments were then dehydrated in solutions of increasing concentrations of ethanol, diaphanized by xylene, to be later impregnated and, finally, included in paraffin. Cuts of 5 micrometers were obtained using a Spencer 820 rotative microtcme from the Psner;-can Optical Oo., and after being mounted on histological slides, were dyed by either the Hematoxillin-Eosin techn,~que, or by the Gomori trichromic method. The preparation could then be examined by an Optiphot model Nikon microscope, coupled to an automatic AFX II
photometer with a 35mm camera, thus obtainng photomicrographies on 100 ASA Ektachrome film from Kodak.
Samples were obtained of tissues from rats in the 1", 2na, 4th, 8" and l5tr weeks, to evaluate possible differences in tissue reactions resulting from the injection of carboxy glyconate hydrolactic of magnesium.
It was verified that the injection of carboxy glyconate hydrolactic of magnesium in albino rats promotes the formation of a aranuloma in reaction to foreign bodies, localized preferably in the area of the injection, that is, in the inferior skeletal muscular layer (present in the rat) of the hypodermic In some situations, however, it reaches the supra muscular region of the hypodermis, attaining deep dermis. This granuloma becomes stable around the second week, after the injection, and the carboxy glyconate hydrolactic magnesium colloid can be visualized in an apparently stable form surrounded by giant foreign body cells. After the eighth week, the granulomas becomes surrounded by conjunctive capsules that extend septa inwards, turning the granulomas lobed. The presence of neoplasia was not observed.
Through photomicrographies, it was possible to observe that when employing the composition of the present invention, collections of droplets of carboxy giyconate hydrolactic magnesium colloid, of irregular shape and showing basophilic content, were observed, either grouped between macrophages or surrounded by giant foreign body cells.
The injection of carboxy glyconate hydrolactic of magnesium colloid produced inflammatory reactions at the end of the first week filling the area with macrophages infiltrating between the droplets of amorphous and basophilic colloid, promoting a matrix deposit well visible below the muscular layer, with an abundance of reticular fibers, revealed by the green refringency, comparable to that of the peri and endomysium.
In the second week, macrophages already form a thick layer around the collections of colloid droplets and infiltrate between colloid conglomerates, forming septa that show more extensive deposits of extracellular matrix rich in reticular fibers in the more slender ones and collagen fibers in the thicker ones.

In the fourth and eighth weeks, it is possible to observe, with clarity, that when of carboxy glyconate hydrolactic of magnesium is injected, amorphous spaces occur containing basophilic material (colloid) forming separate aggregates. In the fourth week and the eighth week, the predominance of fibers with green birefringency in the interstices of the aggregates of the colloid droplets is observed, compatible with the predominance of collagen III, particular to reticular fibers.
In the sixteenth week, it is observed that the colloid granulomas show a predominance of green birefringency in the interstice with slenderer septa and yellow or red birefringency in the thicker septa and capsules. This is in accordance with the greater quantity of collagen III present in the interstice of the smaller septa.
Conclusion of Experiment II
From the observations of Experiment II, it is possible to conclude that the injection of carboxy glyconate hydrolactic of magnesium, in the hypodermis of albino rats, promotes the formation of granulomas reactional to foreign bodies having the same characteristics as extracellular matrix. The granulomas thus constituted become stable around the second week after the injection and, the colloid can be visualized in an apparently stable form surrounded by giant foreign body cells. Already, in the first week, the actual macrophages when infiltrating the colloid begin to promote the septation of the future granuloma, with an initial production of extracellular matrix. The granuloma, in the second week, exhibits more advanced septation and, in the 8t'~' week, possesses a thick conjunctive capsule rich in collagen I that sends septa inwards, making the mentioned granuloma distinctly lobulate. However, inside the lobules, the richness of collagen III is notable. This richness 5 of collagen III inside the stabilized granuloma tends to indicate that the slender conjunctive fibers present there do not characterize a state of fibrosis, when thick conjunctive fibers (collagen fibers) rich in collagen I would predominate.
METHOD FOR TREATMENT OF DERMATOI~OGICAZ DISORDERS
10 A method to treat dermatological disorders by filling and correcting unaesthetical alterations with the composition of carboxy glyconate hydrolactic of magnesium, in accordance with the present invention, is described below.
A composition is obtained by the process already described 15 herein, consisting of mixing all the ingredients in stainless steel tubes, followed by agitation. The temperature of the mixing may vary between 30°C and 7C°C, preferably at around 50°C, and for a time period for approximately 3 hours.
This procedure is repeated twice more, until obtaining the final product.
At this stage a colloidal solution is obtained.
The resulting colloidal product is then filtered, and its pH is verified so as to present a value of approximately 6.0 and a density equal to 1.
The resulting product, that is, the compositon of carboxy glyconate hydrolactic of magnesium, already having the pH and density required, is injected into vials of 20m1.

For the injection of the product, it is initially necessary to identify the areas to be treated and, if necessary, to mark these areas in advance with a patient standing upright. Next, the patient is placed in dorsal decubitus, in a seated position, at an angle of between 45 and 60 degrees, when the area to be treated is the ventral part of the head, or without ventral decubitus, when the area to be treated is the dorsal area.
The asepsis and antisepsis is then performed with the usual antiseptics.
As routine, a blocking or ever. local infiltratio.~.~. with anesthetics is performed for greater comfort and less stress for the patient during the procedure.
There is no necessity of prior tests as the product is inert and biocompatible, that is, exempt of any allergic or toxic reaction. The formulation of the compositio::, in accordance with the present invention, impedes contamination by fungi or bacteria.
The content of the vial is aspirated with a needle or cannula, this latter with a rhombic point. The size of the needle or cannula varies in accordance with the quantity necessary for filling the area to be treated.
The application of the composition of carboxy glyconate hydrolactic of magnesium is performed with a needle or a cannula, depending on the area to be treated.

The injection of the composition should' ~voicy '~hg formation of sequential bubbles, like a pearl necklace, because in this manner the result would be irregular.
Being a permanent implant, hypocorrections were performed and, when necessary, more than one or two complementary sessions can occur, with an interval of 25-30 days between each session. To obtain a better result in total safety, this time interval was established for a further injection. This interval is fundamental for the deposit of collagen and fibrin fibers, providing the necessary support for the microimplant.

Claims

1. Colloidal composition for esthetic corrections characterized by being composed of a colloidal formulation termed carboxy glyconate hydrolactic of magnesium, consisting of:
(a) approximately 5 to 70g of an ether of cellulose;
(b) approximately 800 to 1300ml of Ringuer lactate;
(c) approximately 2 to 20g EDTA;
(d) 20 to 120ml of calcium glyconate; and, (e) optionally, between 0 and 10% xylocaine.

2. Colloidal composition in accordance with Claim 1, characterized by the fact that the ether of cellulose is a polymers of ether of cellulose selected from hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxybutyl cellulose, hydroxyproplymethyl cellulose, carboxymethyl cellulose of sodium, carboxymethyl cellulose of calcium, methylcellulose and/or ethylcellulose.

3. Colloidal composition in accordance with Claim 1 characterized by the Ringuer lactate being an inert solution composed of sodium chloride, potassium chloride, dehydrated calcium chloride, sodium lactate and water.

4. Composition in accordance with Claim 1 characterized by being applied, in humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight.

5. Process for the preparation of the colloidal composition for esthetical correction characterized by including the stages of:

(a) mix an ether of cellulose, a Ringuer lactate solution, EDTA and calcium glyconate;

(b) agitate the ingredients of stage (a), at a temperature varying between 30°C and 70°C, for a time period for approximately 3 hours;

(c) repeat the procedures of stage (b) twice;

(d) filter the product obtained in stage (c, and (e) adjust the pH of the filtered product to an approximate value of 6Ø

6. Process in accordance with Claim 5 characterized by also including the addition of xylocaine, as an optional ingredient.