Ligament regeneration injection compositions for repositioning sutures and uses thereof

Abstract

The present invention relates to a sutured ligament regeneration injection composition for reduction and use thereof. The injection composition of the present invention is prepared in an optimal mixing ratio that maximizes the interaction and effect of each component, thereby more effectively promoting the regeneration and recovery of ligaments, particularly showing excellent effects on the recovery of elbow ligament and wrist ligament and shoulder ligament injuries such as rotator cuff. In addition, the injection composition of the present invention can be applied to a non-invasive treatment method, and compared to the conventional surgical treatment method, the recovery period is shorter, and the burden on the patient can be reduced. Finally, each component of the composition of the present invention has a biocompatible property, so that it can be safely applied to the body, and thus has less side effects and minimizes the risk of infection and complications compared to the conventional surgical method.

Description

Technical Field

[0001] This invention relates to a ligament regeneration injection composition for reduction and suturing and its uses. The injection composition of this invention is prepared by an optimal mixing ratio that maximizes the interaction and effect of the components, and can be used to regenerate ligaments in cases of various ligament ruptures, such as elbow ligaments, wrist ligaments, and rotator cuff (shoulder ligament). Background Technology

[0002] Reduced sutures refer to a method of suturing the rotator cuff using anchors, a procedure previously only possible through arthroscopic surgery. On the other hand, ligaments are vital structures connecting bones throughout the body, maintaining joint stability and supporting range of motion. However, ligaments are frequently damaged or ruptured due to trauma, overuse, and aging. Shoulder ligament tears, elbow ligament tears, and wrist ligament tears (such as rotator cuff injuries) are common not only among athletes but also among the general population. This can lead to pain and limited function, negatively impacting quality of life.

[0003] Currently, ligament regeneration methods include physical therapy, drug injections, and surgery, but each has its limitations. Physical therapy has a long treatment period, drug therapy has limited effectiveness, and surgical methods require a postoperative recovery period and carry risks of complications such as infection. Therefore, new treatment methods are needed for more effective and safer ligament regeneration.

[0004] Recent biotherapies, particularly stem cell therapy and platelet-rich plasma (PRP), have shown positive effects on ligament regeneration; however, these therapies remain expensive and require complex formulations. Furthermore, the instability or difficulty in maintaining effective concentrations of the bioactive components limits their clinical use.

[0005] This invention aims to overcome these existing limitations and to prepare injectable compositions for ligament regeneration through an optimal mixing ratio that maximizes the interaction and effect of each component. The injectable compositions of this invention provide a promoting effect on regeneration in cases of ligament ruptures such as those in the elbow and wrist, as well as shoulder ligament ruptures such as the rotator cuff, and can suggest a safe and effective treatment regimen. Therefore, this invention will provide an innovative solution for ligament regeneration, which will contribute to improved overall quality of life by relieving pain and promoting functional recovery in patients.

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Korean Patent No. 10-2019-0065079 Summary of the Invention

[0009] The problem that the invention aims to solve

[0010] The purpose of this invention is to provide an injectable composition for ligament regeneration and for repositioning sutures.

[0011] Another object of the present invention is to provide a method for preparing an injectable composition for ligament regeneration for repositioning sutures.

[0012] Methods for solving problems

[0013] To achieve the above objectives, the present invention provides a pharmaceutical composition for ligament regeneration, comprising hydrolyzed collagen, hyaluronic acid, fibroblast growth factor (FGF), polydeoxyribonucleotide (PDRN), aloe extract, vitamin C, methanesulfonylmethane (MSM), epidermal growth factor (EGF), glucosamine, silymarin, and physiological saline (N / S).

[0014] In one embodiment of the present invention, the "composition" is a mixture of 20-30g hydrolyzed collagen, 2-3g hyaluronic acid, 0.01-0.1g FGF (fibroblast growth factor), 2-3g PDRN (polydeoxyribonucleotide), 5-15g aloe vera extract, 0.5-1.5g vitamin C, 5-15g MSM (methylsulfonylmethane), 0.01-0.1g EGF (epidermal growth factor), 1-10g glucosamine, and 0.5-1.5g silymarin. The preferred ingredients in 400–600 cc of physiological saline (N / S) are 25 g hydrolyzed collagen, 2.5 g hyaluronic acid, 0.05 g FGF (fibroblast growth factor), 2.5 g PDRN (polydeoxyribonucleotide), 10 g aloe extract, 1 g vitamin C, 10 g MSM (methylsulfonylmethane), and 0.05 g EGF (epidermal growth factor). It can be prepared by mixing 5 g glucosamine and 1 g silymarin with 500 cc of physiological saline (N / S), but is not limited to these ingredients.

[0015] In one embodiment of the present invention, the "composition" may be used to increase the collagen density of ligament tissue, increase the thickness and strength of ligaments, but is not limited to that.

[0016] In one embodiment of the present invention, the "composition" may have a ligament regeneration effect on injuries to the shoulder ligament, elbow ligament or wrist ligament, but is not limited thereto.

[0017] Invention Effects

[0018] This invention relates to a ligament regeneration injection composition for reduction and suturing, and its uses. The injection composition of this invention is prepared with an optimal mixing ratio that maximizes the interaction and effect of each component, thereby more effectively promoting ligament regeneration and recovery, particularly showing excellent results in the recovery of elbow and wrist ligaments, as well as shoulder ligament injuries such as the rotator cuff. Furthermore, the injection composition of this invention can be used in non-invasive treatment methods, resulting in a shorter recovery period compared to traditional surgical methods, reducing the burden on patients. Finally, each component of the composition of this invention has biofriendly properties, making it safe for application to the body, thus resulting in fewer side effects and minimizing the risk of infection and complications compared to traditional surgical methods. Attached Figure Description

[0019] Figure 1 This is a schematic diagram illustrating a surgical scenario of repositioning and suturing in one embodiment of the present invention.

[0020] Figure 2 These are photographs of the ligaments before and after a reduction and suturing procedure using the injectable composition of the present invention in one embodiment of the invention. Detailed Implementation

[0021] However, the following embodiments are given as examples of the present invention. If it is determined that a detailed description of a known technique or composition may unnecessarily obscure the essential points of the invention, the invention may be omitted, and the invention is not limited thereto. The invention can be modified and applied in various variations and applications within the uniform category described in the patent claims below and as interpreted thereby.

[0022] Furthermore, the terminology used in this specification is for the proper description of preferred embodiments of the invention, and these terms may vary depending on the intent of the user, the author, or practice in the art to which this invention pertains. Therefore, the definitions of these terms should be based on the entirety of the specification. Throughout the specification, when a section “includes” a component, it means that other components may be included, rather than excluded, unless specifically stated otherwise.

[0023] In this specification, unless otherwise stated, the "%" used to express the concentration of a particular substance is (w / w)% for solid / solid, (w / v)% for solid / liquid, and (v / v)% for liquid / liquid.

[0024] In terms of application, the present invention provides an injectable pharmaceutical composition for ligament regeneration.

[0025] The pharmaceutical compositions of the present invention may include additional adjuvants in addition to the active ingredient. Adjuvants may be used without limitation, provided they are known in the art, but their function may be enhanced, for example, by including more complete or incomplete Freud supplements.

[0026] The pharmaceutical compositions according to the present invention can be prepared in the form of active ingredients incorporated into a pharmacologically permissible carrier. Here, pharmacologically permissible carriers include carriers, excipients, and diluents commonly used in the pharmaceutical industry. Pharmacologically permissible carriers usable in the pharmaceutical compositions of the present invention include, but are not limited to, lactose, glucose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil.

[0027] Each pharmaceutical composition of the present invention can be used as a sterile injection solution by formulation in accordance with conventional methods.

[0028] In the preparation process, diluents or excipients can be used, such as fillers, fillers, binders, humectants, disintegrants, and surfactants. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Parenteral formulations, such as injectable compositions, can include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents, suspensions, vegetable oils (such as propylene glycol), polyethylene glycol, vegetable oils (such as olive oil), and injectable esters (such as ethyl oleate) can be used. Suppositories can include witepsol, tween61, cacaoji, lauringe, and glycerogelatin. Furthermore, pharmaceutical compositions according to the invention can be administered to an individual via injection into the fat layer of the skin.

[0029] The dosage of the pharmaceutical composition according to the invention is selected taking into account the individual's age, weight, sex, and physical condition. Clearly, the concentration of the active ingredient contained in the pharmaceutical composition can be selected in various ways depending on the target population, and injectable formulations may use fatty acids, such as oleic acid.

[0030] In terms of application, the ligament regeneration injection pharmaceutical composition of the present invention for repositioning sutures can be prepared by mixing hydrolyzed collagen, hyaluronic acid, fibroblast growth factor (FGF), polydeoxyribonucleotide (PDRN), aloe extract, vitamin C, methanesulfonylmethane (MSM), epidermal growth factor (EGF), glucosamine, and silymarin in physiological saline (N / S).

[0031] The roles of the above-mentioned components in the drug composition of the ligament regeneration injection for repositioning and suturing are as follows.

[0032] The composition “HydrolyzedCollagen” used in this invention is a major structural component of ligaments and may play a role in promoting tissue repair and ligament regeneration.

[0033] The composition "hyaluronic acid" used in this invention plays a role in maintaining tissue moisture and relieving inflammation, and can promote tissue regeneration.

[0034] The composition “FGF (fibroblast growth factor)” used in this invention can stimulate the proliferation of fibroblasts to promote the rapid regeneration of ligaments and tissues.

[0035] The composition “PDRN (polydeoxyribonucleic acid)” used in this invention can promote cell regeneration, relieve inflammation and help repair damaged tissue.

[0036] The composition "aloe vera extract" used in this invention has sedative and anti-inflammatory effects, which help with tissue regeneration and inflammation relief.

[0037] The composition “Vitamin C” used in this invention promotes collagen synthesis and can exert antioxidant activity during tissue repair.

[0038] The composition “MSM (methylsulfonylmethane)” used in this invention has excellent anti-inflammatory properties and can help repair ligaments and joint tissues.

[0039] The composition “EGF (epidermal growth factor)” used in this invention can accelerate tissue recovery by promoting cell growth and regeneration.

[0040] The composition "glucosamine" used in this invention helps repair cartilage and ligaments and can increase the flexibility and strength of tissues.

[0041] The composition “silymarin” used in this invention can prevent damage to ligament tissue and promote regeneration through its antioxidant and anti-inflammatory effects.

[0042] However, the embodiments and experimental examples are presented as examples of the present invention. If it is determined that a detailed description of known techniques or compositions may unnecessarily obscure the essential points of the present invention, a detailed description may be omitted, and the present invention is not limited thereto.

[0043] <Production Example> Preparation of Injectable Compositions

[0044] To prepare the ligament regeneration injection composition for repositioning sutures used in this invention, the composition is as follows.

[0045] The composition used in the above mixture is hydrolyzed collagen, hyaluronic acid, FGF (fibroblast growth factor), PDRN (polydeoxyribonucleic acid), aloe vera extract, vitamin C, MSM (methylsulfonylmethane), EGF (epidermal growth factor), glucosamine, and silymarin.

[0046] To determine the optimal mixing ratio for ligament regeneration using the above configuration, its construction is shown in Table 1 below.

[0047] Table 1

[0048]

[0049]

[0050] Example 1: Ligament regeneration effect test

[0051] 1-1. Collagen density measurement, biomechanical assessment, and inflammatory response assessment

[0052] To test the ligament regeneration effect of the injectable composition of the present invention manufactured in the manufacturing example, a 4-week-old male C57BL / 6 mouse was purchased and placed in two groups, one for each experimental group. Ligament regeneration was performed by inducing injury to the Fuji knee ligament region in the mice. For ligament injury, mice were anesthetized and a minor injury was caused to the ligament through a micro-incision, and the ligament regeneration effect was evaluated based on this injury. Subsequently, 0.2 cc was subcutaneously injected into the injured area of ​​each experimental group, for a total of 3 doses, administered every 2 weeks. Collagen density measurement, biomechanical evaluation, and inflammatory response evaluation were performed as evaluation indicators.

[0053] Collagen density measurement is an indicator of regeneration by evaluating the collagen density of ligament tissue. Tissue samples are collected at 2, 4, and 6 weeks after each injection. The degree of collagen formation is evaluated by immunohistochemical staining, and the collagen density of the tissue samples is quantified by electron microscopy.

[0054] Biomechanical evaluation measures the tensile strength of ligament tissue to assess the degree of ligament regeneration. The strength of the ligament (maximum load and tensile force) is measured by mechanical tensile test, and the regenerated ligament tissue is evaluated by mechanical test at 2, 4 and 6 weeks after each injection.

[0055] Inflammatory response assessment evaluates the inflammatory response in tissues to assess the side effects of the injection, and measures the expression of inflammatory cytokines in tissue samples by ELISA.

[0056] The results for each evaluation indicator are shown in Table 2 below.

[0057] Table 2

[0058]

[0059]

[0060] Example 1 showed that the injected composition achieved a collagen formation rate of up to 95%, confirming excellent tissue regeneration. In terms of tensile strength, it reached 90%, demonstrating the fastest and most robust recovery of damaged ligament tissue with virtually no inflammatory response and very low expression of inflammatory cytokines at the injection site. Therefore, in terms of clinical findings on regenerative effects, its regenerative effect was the most outstanding in the experimental group, recording the best results across all indicators.

[0061] The injectable composition of Example 2 showed slightly lower results than that of Example 1 in terms of collagen density (92%), and excellent ligament regeneration in terms of tensile strength (87%), with very rapid tissue recovery after injection. Inflammatory response and low inflammatory response were observed. Therefore, in terms of clinical outcomes regarding regenerative effects, it showed a slightly lower regenerative effect than Example 1.

[0062] The collagen density of the injectable composition in Example 3 was 75%, which is slightly low for collagen formation. The tensile strength was 70%, indicating a moderate regenerative effect, with tissue recovery being slower than in the other experimental groups. The inflammatory response was confirmed to be moderate; therefore, the clinical outcome of the regenerative effect showed a moderate regenerative effect and relatively low results.

[0063] The injectable composition of Example 4 showed good results with 85% collagen density and relatively high regenerative effect at 82% tensile strength. Inflammatory response was low, but mild side effects were observed at some injection sites. Therefore, the overall clinical findings on the regenerative effect indicate a good regenerative effect.

[0064] Example 5 showed a lower collagen density of 82% in the injected composition, but still exhibited good regenerative effects. Tensile strength was 80%, indicating good ligament regeneration. Inflammatory response was low, but some tissue aggregation occurred after injection. Therefore, the clinical findings on the regenerative effect showed good regenerative efficacy, but the results were moderate between experimental groups.

[0065] Comparative analysis of the injectable composition in Example 1 showed lower collagen formation (70% collagen density) compared to another experimental group. Tensile strength was 65%, insufficient to achieve a regenerative effect, and tissue recovery was slow. Inflammatory response was high, with significantly increased expression of inflammatory cytokines compared to other experimental groups. Therefore, clinical findings on regenerative effects are insufficient.

[0066] Comparative analysis of the injection composition in Example 2 showed the lowest collagen density, at 60%. Regarding tensile strength, the ligament regeneration effect was very low, at 58%, with slow recovery after injection. The inflammatory response was very high, with significant inflammation occurring immediately after injection. Therefore, clinical findings on the regenerative effect indicate an overall insufficient regenerative capacity.

[0067] In other words, it was confirmed that the injectable composition of Example 1 showed the best ligament regeneration effect in all evaluation metrics.

[0068] 1-2. Measurement of ligament thickness, ligament strength, and cell proliferation rate

[0069] After the 1-1 evaluation, mice in each experimental group were euthanized, and ligament tissue from the injured area was removed. The removed ligament tissue was immediately fixed with formalin (10% neutral buffer solution). For tissue slide production, the fixed tissue was encased in paraffin and prepared into slides. The slides were stained with H&E staining (hematoxylin and eosin staining) and then observed under a microscope. For thickness measurement, tissue sections were observed under a microscope (400x magnification), and the thickness (μm) of the ligament tissue was measured using an electronic micrometer. Five points were randomly selected from each sample to obtain the average value.

[0070] In addition, the physical strength of the extracted ligaments was tested using a universal testing machine (UTM). After fixing both ends of the ligament tissue, the force until fracture was measured by applying gradual tension. The tensile speed was set to 5 mm / min as the measurement condition, and each sample was measured three times to obtain the average value. The ligament strength result is expressed as the maximum tensile strength (N) at fracture, and the average value of each group is recorded.

[0071] Finally, ligament tissue was removed from the injection site, cell proliferation rate was measured, cells were isolated, and cell proliferation rate was evaluated by MTT assay. The extracted ligament tissue was treated with collagenase to isolate cells and then cultured. Cells were cultured in 96-well plates at 5 x 10³ cells / well, and then MTT reagent was added to measure cell viability. After three replicates for each sample, absorbance was measured at 570 nm. Based on the measured absorbance, cell proliferation rate (%) was calculated, and the regeneration effect was assessed.

[0072] Table 3 below shows the results.

[0073] Table 3

[0074]

[0075] Table 3 shows the ligament thickness (μm), with Example 1 showing the thickest ligament at 340 ± 10 μm. This represents the optimal result for ligament regeneration. Examples 2 through 5 showed relatively similar values ​​in the range of 315–330 μm, and were lower than those in Example 1. Comparative Examples 1 and 2 showed relatively thinner ligament thicknesses, at 280 ± 10 μm and 290 ± 11 μm, respectively. This indicates a relatively low ligament regeneration effect. In terms of ligament strength (N), Example 1 showed the highest ligament strength at 9.8 ± 0.2 N. This means that ligament repair and regeneration were very strong. Examples 2 through 5 showed relatively lower ligament strengths in the range of 9.4–9.7 N, lower than Example 1, and 8.0 ± 0.2 N and 8.5 ± 0.3 N, respectively, compared to Example 1. This, along with thickness, indicates a reduced regenerative effect in the comparative groups. In terms of cell proliferation rate (%), Example 1 showed the highest cell proliferation rate at 85 ± 4%. This indicates that cells actively proliferate to achieve ligament regeneration. Example 3 showed a cell proliferation rate of 80±6%, Example 4 showed a cell proliferation rate of 83±3%, Example 5 showed a cell proliferation rate of 84±4%, and Example 6 showed a cell proliferation rate of 82±5%, which was lower than that of Example 1, but the overall results were good. Compared with Example 1 and Example 2, which showed a cell proliferation rate of 70±7% and 75±6% respectively, the cell proliferation rate was very low. This indicates that the ligament regeneration effect is weakened.

[0076] In other words, Example 1 showed the best results in terms of ligament thickness, ligament strength, and cell proliferation rate. This indicates that the components of Example 1 are the most effective in ligament regeneration. On the other hand, Examples 2 to 5 showed results lower than Example 1, indicating that there are differences in effectiveness depending on the ratio of each component. Therefore, since the injectable composition of Example 1 showed the best results in all evaluation items, the injectable composition of the present invention is confirmed to be the most effective combination for ligament regeneration.

[0077] As can be seen above, specific embodiments of the present invention have been described in detail. However, those who understand the spirit of the invention can easily propose other reverse inventions or other embodiments falling within the scope of the present invention by adding, modifying, or deleting other components within the same scope of the concept. Therefore, the above embodiments should be understood as exemplary and not as limiting in all respects. The scope of the invention is indicated by the scope of the specific claims described below, rather than by the detailed description above. The meaning and scope of the patent claims, as well as any modifications or variations derived from equivalent concepts, should be interpreted as being included within the scope of the invention.

Claims

1. A ligament regeneration pharmaceutical composition for use in repositioning sutures, wherein, It includes hydrolyzed collagen, hyaluronic acid, fibroblast growth factor, polydeoxyribonucleotides, aloe vera extract, vitamin C, methylsulfonylmethane, epidermal growth factor, glucosamine, silymarin, and physiological saline.

2. The ligament regeneration pharmaceutical composition for repositioning sutures according to claim 1, wherein, The ingredients consist of 20-30g hydrolyzed collagen, 2-3g hyaluronic acid, 0.01-0.1g fibroblast growth factor, 2-3g polydeoxyribonucleotide, 5-15g aloe vera extract, 0.5-1.5g vitamin C, 5-15g methanesulfonylmethane, 0.01-0.1g epidermal growth factor, 1-10g glucosamine, 0.5-1.5g silymarin, and 400-600cc physiological saline.

3. The ligament regeneration pharmaceutical composition for repositioning sutures according to claim 1, wherein, Mix 25g hydrolyzed collagen, 2.5g hyaluronic acid, 0.05g fibroblast growth factor, 2.5g polydeoxyribonucleotide, 10g aloe vera extract, 1g vitamin C, 10g methanesulfonylmethane, 0.05g epidermal growth factor, 5g glucosamine, and 1g silymarin in 500cc of physiological saline.

4. The ligament regeneration pharmaceutical composition for repositioning sutures according to any one of claims 1 to 3, wherein, It increases the collagen density of ligament tissue, thereby increasing the thickness and strength of the ligaments.

5. The ligament regeneration pharmaceutical composition for repositioning sutures according to any one of claims 1 to 3, wherein, It has a ligament regeneration effect on ligament injuries of the shoulder ligament, elbow ligament, or wrist ligament.

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