Filler composition containing nucleic acids and calcium hydroxyapatite

Abstract

The present invention relates to a filler composition comprising nucleic acids and calcium hydroxyapatite.

Description

Technical Field

[0001] The present invention relates to a filler composition containing nucleic acid and calcium hydroxyapatite.

Background Art

[0002] As the interest in the appearance of the face, body, etc. has increased, the number of cosmetic surgeries for improving skin conditions and complementing appearance defects has been increasing. In particular, there has been an increasing interest in cosmetic surgeries for improving skin wrinkles and other appearance conditions. In recent years, for both cosmetic purposes and medical purposes of replacing damaged living tissues, a method of injecting a substance similar to skin tissue, so-called filler, into a desired body part has been widely used. Here, the filler plays a role of compensating for skin defects by securing volume under the skin sunken due to facial wrinkles, scars, etc., and particularly improving the appearance of the skin.

[0003] Generally, a filler composition mainly composed of cross-linked hyaluronic acid is widely used. However, since it has a high viscosity and a low elasticity, it is not only difficult to inject into the skin, but even if it is injected into the skin, it cannot maintain the injected form (shape) for a long time, and has a drawback of a short form maintenance period. In addition, in order to produce a skin filler composition having improved elasticity and viscosity that provide a high volumizing ability, showing a low extrusion force, and being usable for a long time, carboxymethyl cellulose (CMC), a synthetic polymer, may be added (Patent Document 1). However, carboxymethyl cellulose (CMC) has a problem that it causes inflammation when subcutaneously administered to animals, and causes fibrosarcoma at the injection site when repeatedly injected, and has low biocompatibility.

[0004] Since a filler composition having excellent biocompatibility, reasonably long usability, and providing an effective volumizing ability has not been sufficiently developed, the development of an improved filler composition is still required.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] Korean Published Patent Publication No. 10-2018-0038753 [Overview of the project] [Problems that the invention aims to solve]

[0006] The present inventors have confirmed that a filler composition containing nucleic acids and calcium hydroxyapatite exhibits excellent biocompatibility and possesses effective volume-adding ability based on superior physical properties, thereby completing the present invention. [Means for solving the problem]

[0007] The present invention aims to provide a filler composition comprising nucleic acids and calcium hydroxyapatite (CaHAp). [Effects of the Invention]

[0008] The filler composition comprising nucleic acids and calcium hydroxyapatite according to the present invention exhibits excellent biocompatibility and effective volume-adding ability. [Brief explanation of the drawing]

[0009] [Figure 1] This figure shows the properties of nucleic acids and calcium hydroxyapatite at various concentrations. [Figure 2] This figure shows the injection pressure measurement results at various concentrations of nucleic acid and calcium hydroxyapatite. [Figure 3] This figure shows the viscoelasticity measurement results at various concentrations of nucleic acid and calcium hydroxyapatite. [Figure 4] This figure shows the results of a comparative evaluation of the physicochemical properties of nucleic acid and calcium hydroxyapatite compositions and carboxymethylcellulose and calcium hydroxyapatite compositions. [Figure 5] This figure shows the results of a comparative evaluation of the biocompatibility of nucleic acid and calcium hydroxyapatite compositions and carboxymethylcellulose and calcium hydroxyapatite compositions. [Modes for carrying out the invention]

[0010] The present invention will be described in detail below. Note that each description and embodiment disclosed herein applies to other descriptions and embodiments. That is, any combination of the various elements disclosed herein is included in the present invention. Furthermore, the present invention is not limited to the following specific descriptions.

[0011] Furthermore, a person with ordinary skill in the art will be able to recognize and confirm many equivalents of the specific embodiments of the present invention described herein using only ordinary experiments. Moreover, these equivalents are also intended to be included in the present invention.

[0012] One embodiment of the present invention provides a filler composition comprising nucleic acid and calcium hydroxyapatite.

[0013] The nucleic acids of the present invention promote the production of the extracellular matrix (ECM), which is a component between cells, and activate the skin's healing ability within the human body. By doing so, they play a role in restoring the regenerative capacity of the skin itself, which has deteriorated due to aging, and thus can improve the functionality of the skin itself. The nucleic acids of the present invention are polynucleotides (PN), polydeoxyribonucleotides (PDRN), or mixtures thereof, and are used in combination with "DNA fragment mixtures," "DNA fractions," etc.

[0014] In this invention, "polynucleotide" is also called "PN" and refers to a DNA or RNA chain, which is a polymer of nucleotides in which nucleotide monomers are covalently linked together in a chain. In this invention, "polydeoxyribonucleotide" is also called "PDRN" and refers to a type of low molecular weight DNA complex having a specific molecular weight, but is not limited to these. For example, the polynucleotide has a relatively longer nucleic acid length or a larger molecular weight compared to polydeoxyribonucleotide and acts as a physical support, providing cell adhesion, lubrication, and buffering effects, and is used as a raw material for medical devices. Polydeoxyribonucleotide is used as a raw material for pharmaceuticals for cell proliferation and tissue regeneration, but is not limited to these.

[0015] In any of the above-described examples, the nucleic acid of the present invention may have a molecular weight of approximately 1 to 100,000 kDa, 5 to 50,000 kDa, 50 to 10,000 kDa, or 50 to 1,500 kDa.

[0016] In any of the embodiments described above, the nucleic acid of the present invention may be obtained by extraction from the testes or semen of a fish. Specifically, the fish may be a salmonid fish. More specifically, it may be salmon or trout, but is not limited to these.

[0017] In this invention, "calcium hydroxyapatite (CaHAp)" is a substance widely used to correct wrinkles and replenish lost volume. While primarily used for wrinkle removal and facial contouring, its applications are not limited to these. Furthermore, the above term may be used interchangeably with calcium hydroxyapatite, CaHA, CaHAp, etc.

[0018] The inventors of the present invention have confirmed that when nucleic acid and calcium hydroxyapatite are included instead of the general composition of conventional fillers, they function as excellent fillers, and thus have completed the present invention.

[0019] The composition of the present invention may contain nucleic acid and calcium hydroxyapatite at specific contents.

[0020] As an example, the nucleic acid contained in the composition of the present invention is 1 wt% to 5 wt% based on the total weight of the composition, and the content of calcium hydroxyapatite is 5 wt% to 30 wt% based on the whole composition, but is not limited thereto.

[0021] In any of the above-described examples, the weight ratio of the nucleic acid and calcium hydroxyapatite contained in the composition of the present invention is 1:1 to 1:30, but is not limited thereto. In any of the above-described examples, the nucleic acid contained in the composition of the present invention may be contained at a content of 1 wt% to 5 wt% based on the total weight of the composition.

[0022] Specifically, the nucleic acid contained in the composition of the present invention is about 1 wt% to 5 wt%, about 1 wt% to 4 wt%, about 1 wt% to 3 wt%, about 1 wt% to 2 wt%, about 2 wt% to 5 wt%, about 2 wt% to 4 wt%, about 2 wt% to 3 wt%, about 3 wt% to 4 wt%, about 3 wt% to 5 wt%, or about 4 wt% to 5 wt% based on the total weight of the composition, but is not limited thereto.

[0023] In any of the above-described examples, the calcium hydroxyapatite contained in the composition of the present invention may be contained at a content of 5 wt% to 30 wt% based on the total weight of the composition.

[0024] Specifically, the calcium hydroxyapatite contained in the composition of the present invention is approximately 5-30% by weight, approximately 5-25% by weight, approximately 5-20% by weight, approximately 5-15% by weight, approximately 5-10% by weight, approximately 10-30% by weight, approximately 10-25% by weight, approximately 10-20% by weight, or approximately 10-15% by weight, based on the total weight of the composition, but is not limited to these amounts.

[0025] In any of the above-described examples, the weight ratio of nucleic acid to calcium hydroxyapatite contained in the composition of the present invention may be 1:1 to 1:30.

[0026] Specifically, the weight ratio of nucleic acids to calcium hydroxyapatite contained in the composition of the present invention is approximately 1:30, 2:30, 3:50, 5:30, 7:30, 10:30, 15:30, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:20, 1:25, or 1:30, but is not limited to these.

[0027] The term "about" as used above includes not only the exact number that follows the term "about," but also a range that is close to or near that number. Considering the context in which the number is used, it is possible to determine whether it is close to or near the specific number mentioned. For example, "about" indicates a range of -10% to +10% of a given number. Another example is that "about" indicates a range of -5% to +5% of a given number. Yet another example is a range that includes, but is not limited to, ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc.

[0028] The filler composition according to the present invention may not contain hyaluronic acid or carboxymethylcellulose.

[0029] Hyaluronic acid fillers are currently the most widely used fillers because they are composed of polysaccharides similar to the components of the human body, resulting in fewer side effects such as skin hypersensitivity reactions, easy surgical and removal procedures, excellent viscoelasticity, and the ability to maintain skin volume and elasticity by retaining skin moisture. Recently, there has been much research into inducing cross-linking of hyaluronic acid to increase particle size and molecular weight, thereby extending the duration of the effect. However, the duration of the effect is relatively short, ranging from 6 to 12 months, which necessitates the inconvenience of repeated surgeries at short intervals.

[0030] Fillers made from synthetic polymers are broken down in the human body over a very long period of time, and are therefore valued and used as semi-permanent fillers compared to hyaluronic acid fillers, which are absorbent fillers. Synthetic polymer fillers are microparticle-shaped fillers and must be administered suspended in a gel carrier such as carboxymethylcellulose (CMC). Depending on the average particle size and size distribution of the microparticles, occlusion often occurs when injecting them into the skin, requiring the use of a thicker needle. In addition, it takes 6 to 8 weeks for the skin to recover after injection, which is a disadvantage as it results in lower patient satisfaction compared to hyaluronic acid fillers, which provide immediate effects after surgery.

[0031] The filler composition according to the present invention has excellent biocompatibility and may be used for skin tissue repair.

[0032] Specifically, the filler composition of the present invention is a composition containing nucleic acids, which are biocompatible substances, namely polynucleotides (PN) or polydeoxyribonucleotides (PDRN). Since nucleic acids are known to have a base sequence ratio similar to that of the human body, unlike conventional carboxymethylcellulose, which causes inflammation when administered subcutaneously to animals and causes fibrosarcoma at the injection site with repeated injections, and thus has low biocompatibility, nucleic acids have high biocompatibility with the human body. Furthermore, by promoting the production of the extracellular matrix (ECM), which is a component between cells, and playing a role in activating the skin's healing ability in the human body, it has the function of restoring the regenerative ability that has deteriorated due to aging of the skin itself, thereby improving the functionality of the skin itself. In addition, because nucleic acids have long chain lengths and large molecular weights and act as physical supports, they have the advantage of maintaining good dispersion of calcium hydroxyapatite particles.

[0033] The filler composition according to the present invention may also complement the volume loss of the calcium hydroxyapatite filler.

[0034] Furthermore, the filler composition according to the present invention may be for dermal injection, which is injected to improve wrinkles and shape the skin.

[0035] Specifically, it is used to improve, but is not limited to, skin wrinkles, glabellar lines, nasolabial folds, chin folds, marionette lines, jawlines, buccal commissures, perioral wrinkles, crow's feet, skin depressions, scars, temples, subdermal support of eyebrows, cheekbones and cheek fat pads, tear troughs, nose, lips, cheeks, chin, around the mouth, infraorbital region, and facial asymmetry.

[0036] The filler composition according to the present invention may further contain at least one compound selected from the group consisting of anesthetics, polyols, vitamins, amino acids, metals, antioxidants, and mineral salts. Specifically, the polyol is glycerin, but is not limited thereto. The polyol is included in the filler composition in an amount of about 0.01 to 30% by weight, about 0.5 to 25% by weight, about 1 to 20% by weight, about 1 to 30% by weight, about 5 to 25% by weight, about 10 to 20% by weight, or about 15% by weight, based on the total weight of the composition, but is not limited thereto.

[0037] The filler composition according to the present invention may contain a buffer, such as a phosphate buffer, to adjust the pH. Since the filler composition according to the present invention is intended to be inserted into the human body, the pH is generally in the range of 6.5 to 7.5, specifically in the range of pH 6.8 to 7.4, but is not limited thereto. Furthermore, the osmolality is about 200 mOsmol / l to about 400 mOsmol / l, specifically about 280 mOsmol / l to about 330 mOsmol / l, but is not limited thereto.

[0038] The filler composition according to the present invention provides a filler composition exhibiting an appropriate range of elastic modulus or viscosity modulus that can be injected into the target tissue.

[0039] In this invention, "elasticity" refers to the properties of an object as a solid when force is applied, that is, the property of changing shape when force is applied but returning to its original shape when the force is removed. The elasticity of a filler refers to the degree to which it maintains the desired shape under skin tissue, and the higher the elasticity, the longer the initial shape can be maintained. Such elasticity is expressed by the storage modulus (G'), and its unit is Pascal (Pa). In this invention, "viscosity" refers to the properties of a liquid, that is, the appearance of a flow with viscosity, which is resistance to flow. Such viscosity is expressed by the loss modulus (G'': viscous modulus), and its unit is Pascal (Pa).

[0040] The higher the elasticity of the filler composition, the more it becomes particle-like and feels slightly harder, which is good for giving a sense of volume and has the advantage of maintaining its shape after the procedure. Also, the higher the elasticity, the longer the filler lasts, but if the elasticity is too high, the surface may not be uniform, and depending on the skill level of the surgeon, layers may form in the skin.

[0041] If the filler composition has high viscosity, it becomes sol-like, soft, and squishy, ​​and blends naturally into the skin, but the relatively firm feeling is reduced, so there are limits to how much volume it can provide.

[0042] Therefore, it is important to inject the target filler into the target tissue while adjusting its elasticity and viscosity.

[0043] In one embodiment, the filler composition of the present invention exhibits an elastic modulus of about 20 Pa to about 45,000 Pa. For example, the composition exhibits an elastic modulus of about 20 Pa to about 45,000 Pa, about 30 Pa to about 30,000 Pa, about 40 Pa to about 20,000 Pa, about 50 Pa to about 10,000 Pa, about 100 Pa to about 8,000 Pa, about 500 Pa to about 7,000 Pa, about 1,000 Pa to about 6,000 Pa, about 1,500 Pa to about 5,000 Pa, and about 2,000 Pa to about 4,000 Pa. In other embodiments, the filler composition of the present invention is used in pressures of approximately 20 Pa or more, approximately 25 Pa or more, approximately 50 Pa or more, approximately 75 Pa or more, approximately 100 Pa or more, approximately 125 Pa or more, approximately 150 Pa or more, approximately 175 Pa or more, approximately 200 Pa or more, approximately 300 Pa or more, approximately 400 Pa or more, approximately 500 Pa or more, approximately 600 Pa or more, approximately 700 Pa or more, approximately 800 Pa or more, approximately 900 Pa or more, approximately 1,000 Pa or more, approximately 1,200 Pa or more, approximately 1,400 Pa or more, approximately 1,600 Pa or more, approximately 1,800 Pa or more, approximately 2,000 Pa or more, approximately 2, It exhibits an elastic modulus of 500 Pa or more, approximately 3,000 Pa or more, approximately 3,500 Pa or more, approximately 4,000 Pa or more, approximately 4,500 Pa or more, approximately 5,000 Pa or more, approximately 5,500 Pa or more, approximately 6,000 Pa or more, approximately 6,500 Pa or more, approximately 7,000 Pa or more, approximately 7,500 Pa or more, approximately 8,000 Pa or more, approximately 8,500 Pa or more, approximately 9,000 Pa or more, approximately 9,500 Pa or more, approximately 10,000 Pa or more, approximately 20,000 Pa or more, approximately 30,000 Pa or more, approximately 40,000 Pa or more, or approximately 45,000 Pa or more.

[0044] Generally used filler compositions have an elastic modulus in the range of approximately 20 Pa to 45,000 Pa. Within this elastic range, a uniform injectable composition is obtained, so nucleic acids and / or calcium hydroxyapatite remain uniform in the tissue within the skin after administration, and there are no differences in effect depending on the site. Furthermore, when injected into the body, the tissue repair effect appears uniformly.

[0045] If the elastic range of the filler composition is approximately 20 Pa or less, it cannot maintain its injected shape for a long time, resulting in a short shape retention period. If the elastic range is very high, such as approximately 45,000 Pa or more, it becomes difficult to push the syringe when injecting it into the skin.

[0046] In one embodiment, the filler composition of the present invention exhibits a viscosity coefficient of about 1 Pa to about 8,000 Pa. For example, the composition exhibits viscosity coefficients of about 3 Pa to about 7,500 Pa, about 5 Pa to about 7,000 Pa, about 10 Pa to about 6,500 Pa, about 20 Pa to about 6,000 Pa, about 30 Pa to about 5,000 Pa, about 40 Pa to about 3,000 Pa, about 50 Pa to about 2,000 Pa, and about 60 Pa to about 1,000 Pa. In other embodiments, the filler compositions of the present invention exhibit viscosity coefficients of approximately 1 Pa or more, approximately 2 Pa or more, approximately 3 Pa or more, approximately 5 Pa or more, approximately 10 Pa or more, approximately 20 Pa or more, approximately 30 Pa or more, approximately 40 Pa or more, approximately 50 Pa or more, approximately 100 Pa or more, approximately 200 Pa or more, approximately 300 Pa or more, approximately 500 Pa or more, approximately 700 Pa or more, approximately 900 Pa or more, approximately 1,000 Pa or more, approximately 1,500 Pa or more, approximately 2,000 Pa or more, approximately 2,500 Pa or more, approximately 3,000 Pa or more, approximately 3,500 Pa or more, approximately 4,000 Pa or more, approximately 4,500 Pa or more, approximately 5,000 Pa or more, approximately 5,500 Pa or more, approximately 6,000 Pa or more, approximately 6,500 Pa or more, approximately 7,000 Pa or more, approximately 7,500 Pa or more, or approximately 8,000 Pa or more.

[0047] Generally used filler compositions have a viscosity coefficient in the range of approximately 1 Pa to approximately 8000 Pa. Within this viscosity range, the composition becomes soft, pliable, and suitable as a Sol, allowing it to naturally blend into the tissues within the skin after administration.

[0048] If the viscosity range of the filler composition is approximately 1 Pa or less, it will blend with the skin, but its ability to reshape itself will be reduced, so it will not be able to fulfill its role as a filler to add volume. If the viscosity range is very high, such as approximately 8,000 Pa or more, it will be difficult to push the syringe when injecting it into the skin, so excessive force will be applied, making it difficult to inject an accurate amount, fine adjustment of the injection volume will be difficult, and side effects may occur.

[0049] The filler composition of the present invention provides an injectable composition. The term "injectable" means having the properties required for administering the composition to the dermal region of an individual using an injection device having a fine needle. The term "fine needle" means, for example, a needle of 27 gauge or less, and the fine needle can be appropriately selected by those skilled in the art depending on the purpose by adjusting the size of the nucleic acid or calcium hydroxyapatite particles.

[0050] The filler composition of the present invention may be injected using a fine needle. For example, it may be injected using a needle of about 27 gauge, about 30 gauge, or about 32 gauge. Specifically, it may be injected using a needle of 22 gauge or less, 27 gauge or less, 30 gauge or less, or 32 gauge or less. Other examples include, but are not limited to, needles of about 22 gauge to about 35 gauge, 22 gauge to about 34 gauge, 22 gauge to about 33 gauge, 22 gauge to about 32 gauge, about 22 gauge to about 27 gauge, or about 27 gauge to about 32 gauge.

[0051] The filler composition of the present invention may be injected at a speed of 12 mm / min and with an extrusion force of about 60 N, about 55 N, about 50 N, about 45 N, about 40 N, about 35 N, about 30 N, about 25 N, about 20 N, or about 15 N. For example, it may be injected with a 27 gauge needle with an extrusion force of about 60 N or less, about 55 N or less, about 50 N or less, about 45 N or less, about 40 N or less, about 35 N or less, about 30 N or less, about 25 N or less, about 20 N or less, about 15 N or less, about 10 N or less, or about 5 N or less. Furthermore, injection may be performed with a 30-gauge needle at a pressing force of approximately 60N or less, approximately 55N or less, approximately 50N or less, approximately 45N or less, approximately 40N or less, approximately 35N or less, approximately 30N or less, approximately 25N or less, or approximately 20N or less. Injection may also be performed with a 32-gauge needle at a pressing force of approximately 60N or less, approximately 55N or less, approximately 50N or less, approximately 45N or less, approximately 40N or less, approximately 35N or less, approximately 30N or less, approximately 25N or less, approximately 20N or less, approximately 15N or less, approximately 10N or less, or approximately 5N or less, but is not limited to these.

[0052] As described above, the filler compositions of the present invention have a wide range of viscoelasticity and injection pressure. Therefore, considering the phase characteristics, an appropriate filler composition can be selected and used according to the application, the injection site of the target tissue, and personal preference. [Examples]

[0053] The present invention will be described in more detail below with reference to examples and experimental cases. However, these examples and experimental cases are merely illustrative of the present invention, and the present invention is not limited to these examples and experimental cases. (Example 1)

[0054] Preparation of composition Compositions with the compositions shown in Table 1 were prepared using PN (Pharmaresearch, Korea, molecular weight 50-1,500 kDa) and CaHAp (nanografi, Germany). Specifically, predetermined amounts of PBS (using 1M Potassium Phosphate Buffer as the stock solution), glycerin, DW, and PN were weighed and mixed, and then dissolved. A predetermined amount of CaHAp was weighed and added to the dissolved solution, and then mixed. The mixture was then packed into glass syringes. The mixture was sterilized at 121°C for 20 minutes.

[0055] [Table 1] (Example 2)

[0056] Property evaluation of each CaHAp+PN content The properties of the compositions listed in Table 1 were evaluated at various CaHAp+PN content levels.

[0057] Specifically, the properties of the mixed solution placed in a glass vial were observed.

[0058] As a result, as shown in Figure 1, when the PN content was 0.5 wt%, phase separation occurred and mixing was impossible. Similarly, when the CaHAp content was 50 wt%, mixing was also impossible. Furthermore, when the PN content was 7 wt% and the CaHAp content was 5-30 wt%, mixing was possible, but the gel collapsed and did not exhibit appropriate properties.

[0059] These results confirmed that, in terms of properties, the material exhibits appropriate characteristics when the PN content is 1% to 5% by weight and the CaHAp content is 5% to 45% by weight. The absence of phase separation means that the particle dispersion of calcium hydroxyapatite is well maintained and that it is acting as a physical support. (Example 3)

[0060] Injection pressure evaluation for each CaHAp+PN content. Using the compositions in Table 1, we attempted to evaluate the injection pressure at each CaHAp+PN content.

[0061] Specifically, the injection pressure was measured under the conditions and method shown in Table 2.

[0062] [Table 2]

[0063] As a result, as shown in Figure 2, measurement was not possible in the areas indicated by the diagonal lines, but in the samples where measurement was possible, it was confirmed that the range was from a minimum of 0.76 N to a maximum of 65.1 N.

[0064] Specifically, when the PN content was 0.5% by weight, for CaHAp content between 5% and 10% by weight, insignificantly low values ​​indicating inappropriate ease of injection were measured. When the PN content was 0.5% by weight, for CaHAp content between 30% and 50% by weight, it was confirmed that the liquid and solid phases separated due to phase separation depending on the PN content, and in some ranges, the injection pressure was too high to measure. Furthermore, when the CaHAp content was between 45% and 50% by weight, it was confirmed that mixing was impossible in some compositions, and the injection pressure could not be measured. However, when the PN content was between 0.5% and 7% by weight, for CaHAp content between 5% and 50% by weight, it was confirmed that various ranges of injection pressures, from approximately 1N to approximately 65N, were observed, depending on the PN content, allowing the composition to be used as a filler. (Example 4)

[0065] Evaluation of physicochemical properties (viscosity and elasticity) at each CaHAp+PN content level. Using the compositions shown in Table 1, we attempted to evaluate the physicochemical properties (viscosity and elasticity) at various CaHAp+PN content levels.

[0066] Specifically, viscosity and elasticity were measured under the conditions and methods shown in Table 3.

[0067] [Table 3]

[0068] As a result, as shown in Figure 3, when the PN content was 0.5 wt% and the CaHAp content was 5 wt% to 10 wt%, insignificantly low values ​​were measured that did not form a gel, and when the CaHAp content was 45 wt% to 50 wt%, measurement was impossible.

[0069] However, when the PN content was between 1% and 5% by weight, measurement was possible when the CaHAp content was between 5% and 30% by weight, and when the PN content was 7% by weight, measurement was possible up to a CaHAp content of 5% by weight.

[0070] Specifically, when the PN content was 0.5% by weight and the CaHAp content was 5% to 10% by weight, it was confirmed that the viscosity and elastic values ​​were approximately 0.5 Pa or less. However, when the PN content was 0.5% by weight and the CaHAp content was 45% to 50% by weight, it was impossible to measure both the viscosity and elastic values.

[0071] When the PN content was 1% to 5% by weight, and when the CaHAp content was 5% to 30% by weight, the measured elastic modulus was approximately 25 Pa to 43,000 Pa, and the measured viscosity coefficient was approximately 1.5 Pa to 8,000 Pa, as shown in Figure 3.

[0072] This suggests that the filler compositions of the present invention possess various physical properties and injection pressures, and that by adjusting the injection pressure and viscoelasticity according to the elasticity and repair support site in the target tissue, the filler can be injected according to the purpose. (Example 5)

[0073] Comparative evaluation of the physicochemical properties (viscosity and elasticity) of CaHAp+PN. We attempted to compare and evaluate the physicochemical properties of CaHAp+PN compositions with compositions containing other components. As a comparative composition, we selected a CaHAp+CMC composition containing CMC (Carboxymethyl cellulose) as a component instead of PN in the filler composition of the present invention.

[0074] Compositions with the compositions shown in Table 4 were prepared as the CaHAp+PN composition and CaHAp+CMC composition. CaHAp was contained in 10% by weight in both compositions, and PN and CMC were contained in 3% by weight in each composition. Specifically, predetermined amounts of PBS, Glycerin, DW, CMC, and PN were weighed and mixed, and then dissolved. Next, predetermined amounts of CaHAp were weighed and added to each solution, and then mixed. The mixed solutions were filled into glass syringes. The mixed solutions were sterilized at 121°C for 20 minutes.

[0075] [Table 4]

[0076] The viscosity and elasticity of each of the aforementioned compositions were measured under the conditions and methods shown in Table 3.

[0077] As a result, as shown in Figure 4, it was confirmed that the CaHAp+PN composition had higher measured elasticity and viscosity than the CaHAp+CMC composition. Therefore, the experiment confirmed that using PN compared to CMC improves the physical properties when preparing a mixture with CaHAp. This means that, based on the improved physical properties described above, the range of physical properties that can be adjusted according to the target tissue is broadened in the CaHAp+PN composition, and the range in which it can be used for tissue repair is significantly expanded. (Example 6)

[0078] Biocompatibility evaluation of CaHAp+PN To evaluate the biocompatibility of the CaHAp+PN composition, a WST assay was performed on the composition. Furthermore, to compare the biocompatibility of this composition, the same experiment was conducted on the CaHAp+CMC composition, and the measured values ​​were compared.

[0079] The CaHAp+PN composition, CaHAp+CMC composition, and control group composition were prepared using the compositions shown in Table 4 and placed in 50 mL conical tubes. They were then mixed using a voltexer and subsequently heated in a water bath at approximately 68°C. The mixtures were mixed using a voltexer at 30-minute intervals and then sterilized in an autoclave at 121°C for 20 minutes. The properties of the mixtures were then observed, followed by mixing using syringes connected to the voltexer, and the resulting mixture was filled into 1 mL glass syringes.

[0080] [Table 5]

[0081] For the WST assay, a human dermal fibroblast (HDF) cell line was used. The media was removed from the stabilized cells (HDF), washed with DPBS (Dulbecco's Phosphate Buffered Saline), and then the DPBS was removed. The cells were then treated with 2 mL of trypsin-EDTA solution per well and cultured at 37°C for 3 minutes before being harvested. The media was mixed into a conical tube to neutralize the trypsin, and the tube was centrifuged at 1000 rpm for 5 minutes. The supernatant of the solution was removed, the media was added to the cell pellet settled at the bottom, and then mixed uniformly by pipetting. The cell count was then measured using a hemocytometer, and the cells were then transferred to a 96-well plate, with 1 × 10⁶ cells per well. 5The solution was added to each well to reach a cell / well ratio, and incubated at 37°C for 24 hours. In the control group, 100L of the sample was used to treat each well, and incubated at 37°C for 24 hours. As described above, the wells incubated for 24 hours were removed, washed with DPBS, and 10L of WST agent (Ez-Cytox) was added to each well in 100L of culture medium. The solution was then incubated at 37°C for 1 hour to allow the reaction to occur. After that, the absorbance was measured at 450nm using a spectrophotometer. Before measuring the absorbance, the wells were gently shaken for about 1 minute.

[0082] As a result, as shown in Figure 5, the measured values ​​for the CaHAp+PN composition were confirmed to be 1.5 times higher than those for the CaHAp+CMC composition. Therefore, the experiment confirmed that using PN compared to CMC significantly improves biocompatibility when preparing a mixture with CaHAp.

[0083] From the above description, those skilled in the art in the field to which the present invention pertains will understand that the present invention can be implemented in other specific forms without altering its technical idea or essential features. It should be understood that the above embodiments are merely illustrative and not limiting. The present invention should be interpreted as encompassing all modified or altered forms derived from the meaning and scope of the claims and their equivalent concepts, rather than the specification.

Claims

1. A filler composition comprising nucleic acids and calcium hydroxyapatite (CaHAp), The nucleic acid content of the composition is 1% to 5% by weight relative to the total composition, and the calcium hydroxyapatite content is 5% to 30% by weight relative to the total composition. Filler composition.

2. The nucleic acid is a polynucleotide, a polydeoxyribonucleotide, or a mixture thereof. The filler composition according to claim 1.

3. The nucleic acid has a molecular weight of 50 kDa to 10,000 kDa. The filler composition according to claim 1.

4. The nucleic acids mentioned above were isolated from the testes or semen of fish. The filler composition according to claim 1.

5. The aforementioned fish is characterized by belonging to the salmon family. The filler composition according to claim 4.

6. The nucleic acid and the calcium hydroxyapatite are characterized in having a weight ratio of 1:1 to 1:

30. The filler composition according to claim 1.

7. The composition is characterized by not containing hyaluronic acid or carboxymethylcellulose. The filler composition according to claim 1.

8. The filler composition is characterized by its excellent biocompatibility and its use in skin tissue repair. The filler composition according to claim 1.

9. The composition is characterized by compensating for the volume loss of calcium hydroxyapatite filler. The filler composition according to claim 1.

10. The aforementioned composition is for skin injection, to be injected for the purpose of improving wrinkles and shaping the skin. The filler composition according to claim 1.

11. The composition is used to improve skin wrinkles, glabellar lines, nasolabial folds, chin folds, marionette lines, jawlines, buccal commissures, perioral wrinkles, crow's feet, skin depressions, scars, temples, subcutaneous support of the eyebrows, cheekbones and cheek fat pads, tear troughs, nose, lips, cheeks, chin, around the mouth, infraorbital region, and facial asymmetry. A filler composition according to any one of claims 1 to 10.

12. The composition further comprises at least one compound selected from the group consisting of anesthetics, polyols, vitamins, amino acids, metals, antioxidants, and mineral salts. The filler composition according to claim 1.

13. The polyol is glycerin. The filler composition according to claim 12.

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