CN109195642B - Filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid and preparation method thereof - Google Patents
Filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid and preparation method thereof Download PDFInfo
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- CN109195642B CN109195642B CN201680086204.9A CN201680086204A CN109195642B CN 109195642 B CN109195642 B CN 109195642B CN 201680086204 A CN201680086204 A CN 201680086204A CN 109195642 B CN109195642 B CN 109195642B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/258—Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
Abstract
The present invention relates to a filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid, which exhibits an excellent tissue enhancement effect by injecting the composition of the present invention into a site requiring tissue enhancement, increasing the volume of the injected site, and generating new tissue, and a method for preparing the same.
Description
Technical Field
The present invention relates to a filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid and a preparation method thereof.
Background
Anti-aging (anti-aging) refers to various treatments for medical maintenance and maintenance of aging. Recently, as the trend of aging and emphasis on appearance is deepened, income is increased, women who enter the society are increased, the demand for anti-aging is rapidly increased, and in addition, government policies and innovations of biology and medical technology, which pay attention to improvement of quality of life, the quality of anti-aging products and services is also continuously developed (Jinxiu Fang, 2014; Jintaiji, 2015). As the anti-aging industry has proliferated into future emerging industries, the facial aesthetic (facial aestheticists) related markets, including botulism, filling, laser therapy, skinning, etc., have attracted attention.
Filler (filler) is, as the term is used, a "substance for filling the skin" and means a substance capable of replenishing the skin tissue. Cosmetic fillers are products which exert their own volume-maintaining effect without pharmacological action for the purpose of restoring facial firmness, improving contour, and relieving facial wrinkles, and are safe surgeries for non-surgical treatments without skin incisions or surgeries (jinzhengying, 2015; Kang i.g., 2015). Such cosmetic fillers are known as injectable facial implants (injectable facial implants), dermal fillers (dermal fillers), wrinkle improving fillers (wrinkle fillers), facial soft tissue fillers (facial soft tissue fillers) or by a variety of names.
The duration of the effect may vary depending on the characteristics of the product (main component, viscosity, etc.), the difference in the operation (injection amount, injection technique), and the characteristics of each patient. They can be classified into biphasic (biphasic) and monophasic (monophasic) fillers according to their properties. The single-phase filler is present in a viscous paste state due to a large amount of a crosslinking agent mixed therein, and is easy to mold to make the surface smooth, but the durability is slightly poor. The two-phase filler has excellent elasticity and is less deformed, but the surface may be uneven (hongkuhong, 2013).
The cosmetic filler is classified into degradable and non-degradable according to its main component. The degradable filler contains a natural polymer such as collagen (collagen), hyaluronic acid (hyaluronic acid), and hydroxyapatite (hydroxyapatite) as a main component, and has an advantage that it is relatively easy to remove by using an enzyme for decomposing the relevant component. Non-degradable fillers have an advantage of maintaining a sustained effect for a long period of time because they are not absorbed but remain in the body, but have a disadvantage of being difficult to remove after surgery, and polymethyl methacrylate (PMMA), crosslinked polyamide derivatives (crosslinked polyamide derivatives), and the like are used as main components. Some cosmetic fillers contain a trace amount of anesthetic in order to reduce pain during surgery (center for information and technology support of medical instruments, 2015; zhao yang river, et al, 2015).
Examples of side effects of the injected filler include edema, pain, tenderness, numbness, bruise, hematocele, redness, erythema, pigmentation, allergic reactions, itching, pruritis (pruritus), asymmetry, an irregular shape, a prominent lump or nodule (nodule), infection, granuloma (granuloma), vascular blood flow abnormality (vasular) and Tyndall effect (Tyndall effect).
Accordingly, the present inventors have conducted studies on a filling composition for tissue enhancement, and have confirmed that a gel formed by injecting a temperature-sensitive hydrogel composition containing nucleic acid, chitosan and hyaluronic acid into the dermal layer of the skin exhibits a sustained tissue enhancement effect of the skin, thereby completing the present invention.
As a prior art, european patent publication No. 0696210 discloses a filler using a hydrogel based on hyaluronic acid and polydeoxyribonucleotide, and does not describe the chitosan of the present invention, although it is similar to the present invention. In addition, korean granted patent No. 1476381 discloses a wrinkle-improving facial or dermal filler containing a DNA polymer isolated from fish semen or eggs, but does not relate to chitosan and hydrogel at all.
Disclosure of Invention
Technical problem to be solved
The object of the present invention is to provide a filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid and a preparation method thereof.
Means for solving the problems
The present invention relates to a filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid.
The composition may be in the range of 10 to 1000: 1: 10 to 1000 by weight of a composition in which nucleic acid, chitosan and hyaluronic acid are mixed.
The composition may be in the range of 25 to 100: 1: a composition of mixed nucleic acid, chitosan and hyaluronic acid in a weight ratio of 25 to 100.
The composition may be in the range of 1: 5 to 5: 1 weight ratio of nucleic acid and hyaluronic acid.
The nucleic acid may be present in an amount of 0.01 to 3% by weight, based on the total weight of the composition.
The nucleic acid may be deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or a mixture thereof.
The nucleic acid may have a molecular weight of 1kDa to 100000 kDa.
The nucleic acid may have a molecular weight of 10kDa to 10000 kDa.
The nucleic acid may have a molecular weight of 50kDa to 3500 kDa.
The chitosan may be present in an amount of 0.00001 wt% to 0.3 wt%, based on the total weight of the composition.
The chitosan may have a molecular weight of 3kDa to 1000 kDa.
The hyaluronic acid may be present in an amount of 0.01 to 3% by weight, based on the total weight of the composition.
The hyaluronic acid may have a molecular weight of 20kDa to 3000 kDa.
In addition, the present invention relates to a method for preparing a filling composition for tissue enhancement, the method comprising: i) a step of preparing a stock solution of nucleic acid; ii) a step of preparing a chitosan stock solution; iii) a step of mixing and stirring the nucleic acid stock solution of the i) step and the chitosan stock solution of the ii) step; iv) adding a hyaluronic acid raw material to the mixed solution of the nucleic acid and the chitosan in the step iii), wherein the weight ratio of the nucleic acid to the chitosan to the hyaluronic acid is 10 to 1000: 1: a step of mixing and stirring in a manner of 10 to 1000; and v) stirring the mixed solution of the nucleic acid, the chitosan and the hyaluronic acid in the step iv), and reducing the temperature to normal temperature.
The preparation method can comprise the following steps: i) a step of preparing a nucleic acid stock solution by placing a nucleic acid into a buffer solution (buffer), stirring at 70 ℃ to 80 ℃ and dissolving for 1 hour to 3 hours; ii) a step of preparing a chitosan stock solution by dissolving chitosan in an acidic buffer solution; iii) a step of mixing the nucleic acid stock solution of the i) step and the chitosan stock solution of the ii) step, and stirring at 60 ℃ to 80 ℃ for 20 minutes to 1 hour; iv) adding a hyaluronic acid raw material to the mixed solution of nucleic acid and chitosan of step iii), wherein the weight ratio of the mixed solution of nucleic acid, chitosan and hyaluronic acid is 10-1000: 1: a step of mixing in a manner of 10 to 1000 and stirring at 55 to 65 ℃ for 1 to 2 hours; and v) stirring the mixed solution of the nucleic acid, the chitosan and the hyaluronic acid in the step iv) for 2 to 4 hours, and reducing the temperature to the normal temperature.
The present invention is described in detail below.
In the filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid, the weight ratio of nucleic acid, chitosan and hyaluronic acid is preferably 10 to 1000: 1: 10 to 1000, more preferably the weight ratio of nucleic acid, chitosan and hyaluronic acid is from 25 to 100: 1: 25 to 100.
The composition may be in the range of 1: 5 to 5: 1 weight ratio of nucleic acid and hyaluronic acid.
The nucleic acid may be present in an amount of 0.01 to 3% by weight, based on the total weight of the composition.
The nucleic acid may be a nucleic acid having a molecular weight of 1kDa to 100000 kDa. Preferably, it may be 10kDa to 10000kDa, and most preferably, it may be 50kDa to 3500 kDa.
The nucleic acid may be deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or a mixture thereof. Preferably, it may be deoxyribonucleic acid.
In addition, the deoxyribonucleic acid may be an oligonucleotide (oligonucleotide), a polynucleotide (polynucleotide), and a polydeoxyribonucleotide (polydeoxyribonucleotide).
The chitosan may be present in an amount of 0.00001 wt% to 0.3 wt%, based on the total weight of the composition.
The molecular weight of the chitosan is preferably 3kDa to 1000kDa, but is not limited thereto.
The hyaluronic acid may be present in an amount of 0.01 to 3% by weight, based on the total weight of the composition.
The hyaluronic acid may have a molecular weight of 20kDa to 3000 kDa.
The preparation method of the filling composition for tissue enhancement may be a preparation method comprising the steps of i) preparing a nucleic acid stock solution by putting a nucleic acid into a buffer solution (buffer), stirring at 70 to 80 ℃, and dissolving for 1 to 3 hours; ii) a step of preparing a chitosan stock solution by dissolving chitosan in an acidic buffer solution; iii) a step of mixing the nucleic acid stock solution of the i) step and the chitosan stock solution of the ii) step, and stirring at 60 ℃ to 80 ℃ for 20 minutes to 1 hour; iv) adding a hyaluronic acid raw material to the mixed solution of nucleic acid and chitosan of step iii), wherein the weight ratio of the mixed solution of nucleic acid, chitosan and hyaluronic acid is 10-1000: 1: a step of mixing in a manner of 10 to 1000 and stirring at 55 to 65 ℃ for 1 to 2 hours; and v) stirring the mixed solution of the nucleic acid, the chitosan and the hyaluronic acid in the step iv) for 2 to 4 hours, and reducing the temperature to the normal temperature.
Examples of the buffer solution that can be used for preparing the nucleic acid stock solution include, but are not limited to, disodium hydrogen phosphate dodecahydrate (sodium phosphate dibasic hydrochloride), sodium chloride (sodium chloride), magnesium chloride (magnesium chloride), potassium chloride (potassium chloride), phosphate buffered saline (phosphate buffer saline), and HEPES (N- (2-hydroxyethyl) piperazine-N' -2-ethanesulfonic acid), preferably disodium hydrogen phosphate dodecahydrate (sodium phosphate dibasic hydrochloride).
Examples of the acidic buffer solution that can be used for preparing the chitosan stock solution include acetic acid (acetic acid), hydrochloric acid (hydrochloric acid), ascorbic acid (ascorbic acid), lactic acid (lactic acid), and nitric acid (nitric acid), and preferably, acetic acid (acetic acid) is used, but not limited thereto.
The tissue augmentation filling composition comprising nucleic acid, chitosan, and hyaluronic acid may be a temperature sensitive hydrogel. The temperature-sensitive hydrogel refers to a hydrogel that changes phase with sol phase transition (phase transition) into gel (gel) or changes phase into sol.
The filling composition for tissue enhancement can exhibit not only a tissue enhancement effect by injection but also a tissue enhancement effect more sustained than that of a conventional filling composition for tissue enhancement by filling a dermal layer, a subcutaneous tissue, a sarcolemma and a sarcolemma as new tissues, in which new blood vessels and collagen are generated by proliferation of fibroblasts (fibroplasts) around the injection site.
Effects of the invention
The present invention relates to a filling composition for tissue enhancement comprising a nucleic acid, chitosan and hyaluronic acid, and a method for preparing the same, wherein when the filling composition for tissue enhancement of the present invention is injected into an intradermal or subcutaneous site of the skin, it is confirmed that the filling composition for tissue enhancement is gelled, and the volume of the injected site is increased. Further, observation of the histological change of the injection site revealed that a new tissue was generated at the injection site.
Thus, it is expected that the tissue-strengthening filling composition of the present invention, which contains nucleic acid, chitosan and hyaluronic acid, can exhibit an excellent tissue-strengthening effect by being applied to a site where tissue strengthening is required.
Drawings
Fig. 1 shows the results of confirming the elasticity (a), viscosity (B), and viscoelasticity (C) of the skin tissue and the filling composition for tissue enhancement.
Fig. 2 shows the structure of the skin tissue and the site of administration of the filling composition for tissue enhancement.
Fig. 3 shows the observation results of histological change (a) and tissue filling effect (B) at the administration site by administration of the filling composition for tissue enhancement.
Fig. 4 shows the results of confirmation of whether or not the composition remained in the sarcolemma (a) and the epidermis and dermis layers (B) after administration of the filling composition for tissue enhancement.
Detailed Description
Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein, and may be embodied in other forms. Rather, the embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.
< example 1. preparation of filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid >
To prepare the filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid of the present invention, a stock solution (stock solution) of nucleic acid and chitosan was prepared.
A nucleic acid stock solution was prepared by dissolving nucleic acid in 130mM disodium hydrogen phosphate dodecahydrate buffer solution for 2 hours or more with a heated stirrer at 75 ℃.
Chitosan stock solutions were prepared using 90mM acetic acid (acetic acid).
The prepared stock solutions of nucleic acid and chitosan were mixed and stirred in a heated stirrer at 70 ℃ for 30 minutes. The hyaluronic acid raw material was further mixed with a mixed solution of nucleic acid and chitosan, and the mixture was stirred in a heating stirrer at 60 ℃ for 1 hour, and then stirred at room temperature for 3 hours to prepare a filling composition for tissue enhancement containing nucleic acid, chitosan and hyaluronic acid. At this time, mixing was performed in such a manner that the concentrations of nucleic acid, chitosan and hyaluronic acid reached the concentrations as in table 1 below.
[ Table 1]
< comparative example 1 preparation of filling composition for tissue enhancement to be compared >
The filling composition for tissue enhancement of the comparison object was prepared so as to reach the concentration of the comparative example shown in table 2 below. The preparation was carried out in the same manner as in example 1.
[ Table 2]
< Experimental example 1. measurement of viscoelasticity of filling composition for tissue enhancement >
In order to confirm the viscoelasticity of the tissue-reinforcing filler composition of the present invention, which has characteristics related to the duration (duration) and volume (volume), a rheometer (rheometer) was used. In this case, the measurement conditions used were PU20, Gap (Gap)1 mm, 1Hz, 1% stress-strain rate (stress strain), temperature was increased by 1 ℃ every 1 minute from 25 ℃ to 40 ℃, and G' (Pa) (elastic coefficient), G "(Pa) (viscosity coefficient), and G × (Pa) (viscoelastic coefficient) were measured.
As for the viscoelasticity measurement, in order to compare the example 1-5 and the mixed composition in which the inventor's visual inspection and the injection test by syringe filling were carried out to use the most excellent feeling in the example 1, the comparative examples 1-1, 1-2 and 1-3 were selected, and the results of the measurement are shown in FIG. 1, in which the results of the measurement at 36 ℃ are shown in the following Table 3.
[ Table 3]
As shown in fig. 1 and table 3, the compositions of examples 1 to 5 in which nucleic acid, chitosan and hyaluronic acid were mixed exhibited superior values in all of elasticity (fig. 1A), viscosity (fig. 1B) and viscoelasticity (fig. 1C) as compared to the compositions of comparative examples 1 to 1 in which nucleic acid was present alone, comparative examples 1 to 2 in which nucleic acid and hyaluronic acid were mixed, and comparative examples 1 to 3 in which nucleic acid and chitosan were mixed, and it was confirmed that the compositions of examples 1 to 5 were the most suitable combinations for use as filling compositions for tissue enhancement. In addition, when chitosan was added to and mixed with a composition composed of nucleic acid and hyaluronic acid, it was found that the physical properties of the composition were changed.
< Experimental example 2 preparation of Experimental animals >
For animal experiments, male rats (Rat) of the Sprague-Dawley line of 10 weeks of age were purchased from YOUNG BIO corporation (south city, korea), and two rats were used for each experimental group. In the stainless-steel-wire-mesh cultivation box (W × D × H is 260 mm × 305 mm × 210 mm), two feeds were performed during the quarantine and acclimation period, and one feed was performed during the observation period. As the feed, solid feed for laboratory animals (Harlan laboratories, inc., USA) was used, and the solid feed was fed into a feeder to allow free intake. In drinking water, tap water is filtered by a filtered water sterilizer and then irradiated with ultraviolet rays, and is freely taken by an automatic water supply device. For the environment of the experimental animal breeding room, the temperature is kept at 23 +/-3 ℃, the relative humidity is kept at 55 +/-15%, the illumination time is 12 hours (the lamp is turned on at 8 am to turned off at 8 pm), and the illumination intensity is 150-300 Lux.
< Experimental example 3 infusion of filling composition for tissue enhancement >
Rats raised in experimental example 2 were subjected to respiratory anesthesia using Isoflurane (Isoflurane). The back of the anesthetized rat was shaved, the injection site was sterilized with iodine cotton, and 0.5cc of the filling composition for tissue enhancement was injected Intradermally (ID) or Subcutaneously (SC) using a 1cc disposable sterile syringe and a 30 gauge needle as labeled in fig. 2. After the injection of the composition, the composition was fed in an incubator for 1 week, and the effect, biocompatibility, tissue enhancement, and the like of the filling composition for tissue enhancement were observed.
< Experimental example 4. histopathological Observation >
The filling composition for tissue enhancement of the present invention was injected subcutaneously with CO after 7 days2After air-sacrifice of the rats, the tissue of the subcutaneous back site injected with the composition was collected. The collected tissue was fixed with 10% neutral buffered formalin (neutral buffered formalin) for 24 hours or more, dehydrated through a general tissue processing procedure, and then soaked with paraffin. The tissue after completion of the soaking was sliced at a thickness of 5 μm. The prepared tissue sections were reacted in Mayer's hematoxylin line (Sigma, USA) for 1 second, and then washed with running water for 10 minutes.After that, the reaction was carried out in a solution of eosin (Sigma, USA) for 3 seconds. The stained tissue is subjected to a dehydration process
(Fischer scientific, USA) seal. Using a microscope to perform H&The E-stained tissue sections were observed for histopathological changes, and the results are shown in FIG. 3.
As shown in fig. 3, (a) shows the tissue-strengthening effect of the filling composition for tissue enhancement of the present invention injected into the skin after 7 days. When treated with the composition of comparative example 1-1, the tissue enhancement or filling effect was observed in the lower part of the sarcolemma as compared with the untreated group, but the effect was not large. In contrast, when the compositions of comparative examples 1-2 and comparative examples 1-3 were injected, the tissue layers became slightly thicker compared to the case without treatment, but the difference between the two groups was not large. When treated with the compositions of examples 1 to 5, the lower part of the sarcolemma appeared thicker, and new tissue was generated and filled in the place where the composition existed, as compared with the case of untreated and treated with the composition of comparative example. (B) In the case of (3), the tissue augmentation filling composition of examples 1 to 5 exhibited a tissue filling effect after injection. That is, the density of the tissue increases as the tissue-strengthening filler composition fills the cells to form a layer at the site where the tissue-strengthening filler composition is present. This indicates that the filling composition for reinforcing a tissue of the present invention exhibits a tissue-reinforcing effect not by simple gelation but by tissue formation.
From this, it was found that the tissue-strengthening effect of the filling composition for tissue strengthening of the present invention not only increases the volume of the tissue due to gelation of the composition, but also increases the density of the tissue by generating new tissue such as blood vessels and collagen at the site where the composition is injected, thereby further improving the tissue-strengthening effect.
< Experimental example 5 Observation of volume Change caused by injection of filling composition for tissue augmentation >
The result of the injection of the filling composition for tissue enhancement of the present invention into the subcutaneous tissue and the incision of the injection site on the 3 rd or 7 th day after the injection was performed according to the method of the above experimental example 3 to confirm whether the composition remained in the epidermis and the dermis layer or the sarcolemma is shown in fig. 4.
As shown in fig. 4, in (a), the epidermis and the dermis layer or the sarcolemma are separated by cutting the injection site of the filling composition for tissue enhancement, and after fixing the separated sarcolemma with formalin, the tissue is cut in half for observation of the cross section of the tissue. When the sarcolemma was observed, the filling composition for tissue enhancement was injected, and after 3 days, it was confirmed that the volume of the sarcolemma of the rat into which only the filling composition for tissue enhancement of examples 1 to 5 was injected was increased, and as a result of confirming the section of the sarcolemma, it was confirmed that the gel-like composition remained in the filling composition for tissue enhancement. In (B), the tissue-reinforcing filler composition was injected, and after 7 days, whether or not the gel composition remained in the epidermis and dermis layers was observed. As a result, when the filling compositions for tissue enhancement of comparative examples 1-1, 1-2 and 1-3 were injected, no composition remained in the epidermis and dermis layers. On the contrary, in the case where the filling compositions for tissue enhancement of examples 1 to 5 were administered, it was confirmed that transparent gel-like substances remained in the epidermis and the dermis, and thus it was found that the tissue-enhancing effect of the filling compositions for tissue enhancement of the present invention could be maintained for a long period of time.
Claims (14)
1. A filling composition for tissue enhancement comprising a nucleic acid, chitosan and hyaluronic acid,
wherein the weight ratio of nucleic acid, chitosan and hyaluronic acid of the composition is 10 to 1000: 1: 10 to 1000.
2. The tissue augmentation filling composition of claim 1,
the weight ratio of nucleic acid, chitosan and hyaluronic acid of the composition is 25 to 100: 1: 25 to 100.
3. The tissue augmentation filling composition of claim 1,
the weight ratio of nucleic acid and hyaluronic acid of the composition is 1: 5 to 5: 1.
4. the filling composition for tissue enhancement according to claim 1 or 2,
the nucleic acid is present in an amount of 0.01 to 3 wt% based on the total weight of the composition.
5. The filling composition for tissue enhancement according to claim 1 or 2,
the nucleic acid is deoxyribonucleic acid, ribonucleic acid or a mixture thereof.
6. The filling composition for tissue enhancement according to claim 1 or 2,
the nucleic acid has a molecular weight of 1kDa to 100000 kDa.
7. The tissue augmentation filling composition of claim 6,
the molecular weight of the nucleic acid is 10kDa to 10000 kDa.
8. The tissue augmentation filling composition of claim 7,
the molecular weight of the nucleic acid is 50kDa to 3500 kDa.
9. The filling composition for tissue enhancement according to claim 1 or 2,
the chitosan is contained in an amount of 0.00001 wt% to 0.3 wt% based on the total weight of the composition.
10. The filling composition for tissue enhancement according to claim 1 or 2,
the molecular weight of the chitosan is 3kDa to 1000 kDa.
11. The filling composition for tissue enhancement according to claim 1 or 2,
the content of the hyaluronic acid is 0.01-3 wt% based on the total weight of the composition.
12. The filling composition for tissue enhancement according to claim 1 or 2,
the molecular weight of the hyaluronic acid is 20kDa to 3000 kDa.
13. A method for producing a filling composition for reinforcing tissue, characterized in that,
a method for preparing the filling composition for tissue enhancement as set forth in claim 1 or 2, comprising:
i) a step of preparing a nucleic acid stock solution;
ii) a step of preparing a chitosan stock solution;
iii) a step of mixing and stirring the nucleic acid stock solution of the i) step and the chitosan stock solution of the ii) step;
iv) adding a hyaluronic acid raw material to the mixed solution of the nucleic acid and the chitosan of step iii) and mixing, such that the weight ratio of the nucleic acid, the chitosan and the hyaluronic acid is 10 to 1000: 1: a step of mixing and stirring in a manner of 10 to 1000; and
v) stirring the mixed solution of the nucleic acid, chitosan and hyaluronic acid in the step iv), and reducing the temperature to normal temperature.
14. The method of preparing a filling composition for tissue enhancement according to claim 13,
the preparation method comprises the following steps:
i) a step of preparing a stock solution of nucleic acid by placing nucleic acid in a buffer solution, stirring at 70 ℃ to 80 ℃ and dissolving for 1 hour to 3 hours;
ii) a step of preparing a chitosan stock solution by dissolving chitosan in an acidic buffer solution;
iii) a step of mixing the nucleic acid stock solution of the i) step and the chitosan stock solution of the ii) step, and stirring at 60 ℃ to 80 ℃ for 20 minutes to 1 hour;
iv) adding a hyaluronic acid raw material to the mixed solution of nucleic acid and chitosan of step iii), wherein the weight ratio of the mixed solution of nucleic acid, chitosan and hyaluronic acid is 10-1000: 1: a step of mixing in a manner of 10 to 1000 and stirring at 55 to 65 ℃ for 1 to 2 hours; and
v) stirring the mixture of the nucleic acid, chitosan and hyaluronic acid obtained in the step iv) for 2 to 4 hours, and cooling the temperature to normal temperature.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160070452A KR101710639B1 (en) | 2016-06-07 | 2016-06-07 | Filler composition comprising nucleic acid, chitosan and hyaluronic acid for tissue augmentation and process for producing the same |
| KR10-2016-0070452 | 2016-06-07 | ||
| PCT/KR2016/007237 WO2017213285A1 (en) | 2016-06-07 | 2016-07-05 | Filler composition for tissue augmentation comprising nucleic acid, chitosan, and hyaluronate, and preparation method therefor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109195642A CN109195642A (en) | 2019-01-11 |
| CN109195642B true CN109195642B (en) | 2021-12-10 |
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| CN201680086204.9A Active CN109195642B (en) | 2016-06-07 | 2016-07-05 | Filling composition for tissue enhancement comprising nucleic acid, chitosan and hyaluronic acid and preparation method thereof |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR101710639B1 (en) |
| CN (1) | CN109195642B (en) |
| WO (1) | WO2017213285A1 (en) |
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| JP6660469B2 (en) | 2015-11-24 | 2020-03-11 | ビーエムアイ コリア カンパニー リミテッド | Hyaluronic acid injectable composition containing hyaluronic acid derivative and DNA fraction and use thereof |
| KR102132478B1 (en) * | 2018-11-15 | 2020-07-10 | (주)진우바이오 | Hyaluronic Acid-Polydeoxyribonucleotide Complex and Its Film and Preparation Method Thereof |
| KR102274661B1 (en) * | 2020-10-26 | 2021-07-09 | 주식회사 이노테라피 | Biocompatible film and manufacturing method for the same |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6368356B1 (en) * | 1996-07-11 | 2002-04-09 | Scimed Life Systems, Inc. | Medical devices comprising hydrogel polymers having improved mechanical properties |
| WO2002041928A1 (en) * | 2000-11-22 | 2002-05-30 | Assistance Publique - Hopitaux De Paris | Porous polymeric biomaterials, preparation method and uses |
| CN101400327A (en) * | 2006-03-13 | 2009-04-01 | 凯塞拉生物药品公司 | Fluidic tissue augmentation compositions and methods |
| CN101925348A (en) * | 2007-12-07 | 2010-12-22 | 实验室维维西公司 | Biodegradable single-phase cohesive hydrogel |
| CN102068719A (en) * | 2011-01-18 | 2011-05-25 | 复旦大学 | Adhesion prevention material formed by physical crosslinking hydrogel composition and preparation method and application thereof |
| CN103755981A (en) * | 2014-01-14 | 2014-04-30 | 西北大学 | Hydrogel used as skin filling material and preparation method thereof |
| EP2745849A1 (en) * | 2012-12-19 | 2014-06-25 | Idea Medical Devices S.r.l. | Polydeoxyribonucleotides composition and uses thereof |
| KR20140102208A (en) * | 2011-11-09 | 2014-08-21 | 트러스티즈 오브 터프츠 칼리지 | Injectable silk fibroin particles and uses thereof |
| CN105504313A (en) * | 2016-01-22 | 2016-04-20 | 杭州协合医疗用品有限公司 | Preparation method and application of hyaluronic acid plural gel |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2704431B1 (en) | 1993-04-30 | 1995-07-21 | Sebbin Laboratoires | Use of hydrogels based on hyaluronic acid and / or polydeoxyribonucleotides as filling materials for prostheses and prostheses resulting therefrom. |
| US6254634B1 (en) * | 1998-06-10 | 2001-07-03 | Surmodics, Inc. | Coating compositions |
| EP1407759B8 (en) * | 2002-10-11 | 2010-02-24 | Zummer Ventures LLC | A cosmetic composition in colloidal form comprising hyaluronic acids and chitosan |
| JP4354445B2 (en) * | 2005-10-05 | 2009-10-28 | 恵雄 岡畑 | Method for forming DNA / chitosan complex |
| RU2010116775A (en) * | 2007-09-28 | 2011-11-10 | Энджин, Инк. (Ca) | HIGHLY CONCENTRATED COMPOSITIONS OF CHITOSAN POLYLEX AND NUCLEIC ACID |
| KR101476381B1 (en) | 2010-08-09 | 2014-12-24 | 정래준 | Compositions for enhancing activity of cells, and face or skin filler comprising the polynucleotide fragments complex separated from fish's semen or egg |
| EP2508212A1 (en) * | 2011-04-05 | 2012-10-10 | Universitätsklinikum Freiburg | Biocompatible and biodegradable gradient layer system for regenerative medicine and for tissue support |
-
2016
- 2016-06-07 KR KR1020160070452A patent/KR101710639B1/en active IP Right Grant
- 2016-07-05 CN CN201680086204.9A patent/CN109195642B/en active Active
- 2016-07-05 WO PCT/KR2016/007237 patent/WO2017213285A1/en active Application Filing
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6368356B1 (en) * | 1996-07-11 | 2002-04-09 | Scimed Life Systems, Inc. | Medical devices comprising hydrogel polymers having improved mechanical properties |
| WO2002041928A1 (en) * | 2000-11-22 | 2002-05-30 | Assistance Publique - Hopitaux De Paris | Porous polymeric biomaterials, preparation method and uses |
| CN101400327A (en) * | 2006-03-13 | 2009-04-01 | 凯塞拉生物药品公司 | Fluidic tissue augmentation compositions and methods |
| CN101925348A (en) * | 2007-12-07 | 2010-12-22 | 实验室维维西公司 | Biodegradable single-phase cohesive hydrogel |
| CN102068719A (en) * | 2011-01-18 | 2011-05-25 | 复旦大学 | Adhesion prevention material formed by physical crosslinking hydrogel composition and preparation method and application thereof |
| KR20140102208A (en) * | 2011-11-09 | 2014-08-21 | 트러스티즈 오브 터프츠 칼리지 | Injectable silk fibroin particles and uses thereof |
| CN104363931A (en) * | 2011-11-09 | 2015-02-18 | 塔夫茨大学信托人 | Injectable silk fibroin particles and uses thereof |
| EP2745849A1 (en) * | 2012-12-19 | 2014-06-25 | Idea Medical Devices S.r.l. | Polydeoxyribonucleotides composition and uses thereof |
| CN103755981A (en) * | 2014-01-14 | 2014-04-30 | 西北大学 | Hydrogel used as skin filling material and preparation method thereof |
| CN105504313A (en) * | 2016-01-22 | 2016-04-20 | 杭州协合医疗用品有限公司 | Preparation method and application of hyaluronic acid plural gel |
Non-Patent Citations (4)
| Title |
|---|
| Chitosan/hyaluronic acid polyelectrolyte complex hydrogels in the management of burn wounds;Vasile C;《Europe PMC》;20130430;第565-571页 * |
| Injectable in situ forming biodegradable chitosan-hyaluronic;Palumbo F;《Carbohydrate Polymers》;20061231;第3卷(第66期);第379-385页 * |
| 透明质酸复合材料研究现状;丁金聚;《中国组织工程研究》;20151231(第21期);第3403-3408页 * |
| 透明质酸的结构、性能、改性和应用研究进展;张堃;《高分子通报》;20151231;第217-226页 * |
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| CN109195642A (en) | 2019-01-11 |
| KR101710639B1 (en) | 2017-03-08 |
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