Traditional Chinese medicine composition for treating orthopedic soft tissue injury and preparation method thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a traditional Chinese medicine composition for treating orthopedic soft tissue injury and a preparation method thereof. The composition comprises the following raw materials in parts by weight: 4-6 parts of centella asiatica extract, 4-6 parts of salvia miltiorrhiza extract, 4-6 parts of pseudo-ginseng extract, 2-3.5 parts of eucommia ulmoides extract, 2-4 parts of liquorice extract, 4-7 parts of frankincense resin extract, 4-6.5 parts of turmeric extract and 3-4.5 parts of ligusticum wallichii extract. According to the invention, breakthrough is realized through prescription-drug delivery system-process triple synergy, and in prescription, the eight traditional Chinese medicine extracts are in multi-target synergy, and the whole process of anti-inflammation, analgesia and repair promotion is covered. On the delivery system, the hydrophilic component is included by cyclodextrin to be solubilized and stable, and the lipophilic component is efficiently transdermally loaded in the temperature-sensitive gel in the form of nanoemulsion to form a long-acting drug delivery bin on the skin. According to the process, microscopic uniformity and macroscopic stability of the system are ensured through precise low-temperature swelling, split-phase treatment and vacuum integration.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceutical technology, specifically relating to a traditional Chinese medicine composition for treating orthopedic soft tissue injuries and its preparation method. Background Technology

[0002] Orthopedic soft tissue injuries refer to injuries to various tissues below the skin and outside the bones in the human musculoskeletal system, including muscles, tendons, ligaments, synovium, and fascia, caused by acute external force or chronic strain. Common manifestations include contusions, strains, and sprains. The core pathological process of these diseases usually begins with the destruction of the microscopic or macroscopic structure of local tissues, subsequently triggering a series of complex physiological and biochemical reactions. Its characteristics can be summarized as four main symptoms: inflammation, pain, swelling, and functional impairment. Ideally, natural recovery is a body-led, orderly biological process encompassing the acute inflammatory phase, tissue repair phase, and functional remodeling phase, with the ultimate goal of healing damaged structures and restoring normal function. To intervene in and optimize this recovery process, topical medications have become one of the preferred treatment options due to their localized effects and ease of use. Currently, mainstream topical medications can be broadly divided into two categories. One category consists of Western medicine preparations, represented by nonsteroidal anti-inflammatory drugs (NSAIDs). These can effectively relieve pain and inflammation by inhibiting key targets such as cyclooxygenase. However, their mechanism of action is relatively simple, mainly focusing on symptom management, with limited effects on promoting tissue repair and improving microcirculation. Furthermore, long-term or large-area use carries the potential risk of causing skin allergies, dryness, or even systemic adverse reactions, failing to meet the need for comprehensive treatment of injuries. Another major category comprises traditional Chinese medicine topical preparations with a long history, such as traditional plasters, tinctures, and ointments. Traditional Chinese medicine theory emphasizes treating such injuries through a multi-pathway synergy of promoting blood circulation, removing blood stasis, reducing swelling and relieving pain, and relaxing muscles and tendons, theoretically possessing an overall advantage of multi-target regulation. However, most existing traditional Chinese medicine topical dosage forms face significant physical-pharmacological bottlenecks in practical applications. On the one hand, traditional dosage forms generally have low transdermal penetration efficiency for drug components. Many large molecules or lipophilic active ingredients have difficulty effectively penetrating the skin's stratum corneum barrier, making it difficult for the drug to reach the affected area directly. On the other hand, these preparations generally suffer from short drug retention time at the affected area, easily being lost due to limb movement or clothing friction, failing to maintain a sustained and effective local drug concentration, thus affecting the stability and sustainability of the therapeutic effect. In addition, traditional formulation processes lack sufficient control over the solubility, stability, and release behavior of complex traditional Chinese medicine components, which also restricts the full realization of their therapeutic effects. Summary of the Invention

[0003] In view of the shortcomings of the prior art, the purpose of this invention is to provide a traditional Chinese medicine composition for treating orthopedic soft tissue injuries and its preparation method.

[0004] The technical effects described in this invention are achieved through the following technical solution: a traditional Chinese medicine composition for treating orthopedic soft tissue injuries, comprising the following raw materials in parts by weight: 4-6 parts Centella asiatica extract, 4-6 parts Salvia miltiorrhiza extract, 4-6 parts Panax notoginseng extract, 2-3.5 parts Eucommia ulmoides extract, 2-4 parts Glycyrrhiza uralensis extract, 4-7 parts Boswellia carterii resin extract, 4-6.5 parts Curcuma longa extract, and 3-4.5 parts Ligusticum chuanxiong extract; Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 4.5-6 parts Centella asiatica extract, 4.5-6 parts Salvia miltiorrhiza extract, 2.5-3.5 parts Panax notoginseng extract, 2.5-3.5 parts Eucommia ulmoides extract, 3-4 parts Glycyrrhiza uralensis extract, 5-7 parts Boswellia carterii resin extract, 5-6.5 parts Curcuma longa extract, and 3.5-4.5 parts Ligusticum chuanxiong extract; Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 5 parts Centella asiatica extract, 5 parts Salvia miltiorrhiza extract, 3 parts Panax notoginseng extract, 3 parts Eucommia ulmoides extract, 3.5 parts Glycyrrhiza uralensis extract, 6 parts Boswellia carterii resin extract, 6 parts Curcuma longa extract and 4 parts Ligusticum chuanxiong extract; Preferably, in any of the above-described traditional Chinese medicine compositions for treating orthopedic soft tissue injuries, the composition further includes the following raw materials in parts by weight: 180-220 parts poloxamer 407, 20-30 parts poloxamer 188, 1-2 parts high molecular weight hyaluronic acid, 0.3-0.8 parts low molecular weight hyaluronic acid, 18-26 parts hydroxypropyl-β-cyclodextrin, 0.1-1 parts penetration enhancer, 40-60 parts oil phase, 6-12 parts polyoxyethylene hydrogenated castor oil, 35-50 parts Tween-80, 15-20 parts propylene glycol, 3-4 parts phenoxyethanol, 10-15 parts ethanol, and 5-12 parts lecithin; Preferably, the low molecular weight hyaluronic acid has a molecular weight of 30–100 kDa; Preferably, the molecular weight of the high molecular weight hyaluronic acid is 800-1200 kDa; Preferably, the penetration enhancer is any one of oleic acid, menthol, and borneol; more preferably, it is oleic acid. Preferably, the oil phase is either a medium-chain triglyceride or isopropyl palmitate; Preferably, another aspect of the present invention provides a method for preparing a traditional Chinese medicine composition for treating orthopedic soft tissue injuries, specifically comprising the following steps: S1: Slowly add poloxamer 407 and poloxamer 188 to deionized water at 4°C, stir to dissolve evenly, and let stand at 4°C for 8-12 hours to obtain phase A; S2: Slowly add high molecular weight hyaluronic acid and low molecular weight hyaluronic acid to deionized water at 4℃, stir at 200-300 rpm for 30-60 minutes to hydrate evenly, then add part of propylene glycol, stir to dissolve evenly, and obtain phase C; S3: Add hydroxypropyl-β-cyclodextrin to deionized water, stir at 400-600 rpm for 10-15 min, then slowly add Centella asiatica extract, Salvia miltiorrhiza extract, Panax notoginseng extract, Eucommia ulmoides extract and Glycyrrhiza uralensis extract in sequence, stirring for 10 min after each addition of each extract; then filter through 0.45 μm to obtain phase B; S4: Under 60℃ water bath conditions, the oil phase and lecithin were stirred and mixed at 500 rpm for 10-15 min. Then, the penetration enhancer was added and stirred until evenly dissolved. The frankincense resin extract, turmeric extract and chuanxiong extract were dissolved in sequence, and stirred for 5-10 min after each addition. Then, Tween-80, polyoxyethylene hydrogenated castor oil and ethanol were added, and the mixture was stirred at 60℃ for 10 min. Then, the rotation speed was increased to 8000-12000 rpm for shearing treatment for 3-5 min, followed by ultrasonic treatment to obtain phase D. S5: Dissolve phenoxyethanol in an equal weight of propylene glycol to obtain phase E; S6: Place phase A from step S1 in a container and stir continuously at 300 rpm. Then, slowly add phase C from step S2 over 10–15 min, maintaining a material temperature of 5–10°C. Add phase B from step S3 over 20–30 min, maintaining a material temperature of 5–10°C. Add phase D from step S4 in a thin stream over 5–10 min, increasing the rotation speed to 500 rpm and maintaining a material temperature of 5–10°C. Continue stirring for 20 min after adding the phase D. Add phase E from step S5, stir for 10 min, add 10 mM PBS buffer to adjust the pH to 6.4–6.6, and degas under vacuum to obtain the traditional Chinese medicine composition. Preferably, in step S4, the ultrasonic treatment parameters are: 200-400W, 5s on / 5s off, cumulative 3-5min, with the temperature controlled by an ice bath below 25℃ throughout the process; Preferably, in step S6, the vacuum degassing parameters are: -0.08 MPa, time 5 to 10 min, and the process is repeated twice.

[0005] The beneficial effects of this invention are as follows: Compared with existing technologies, the beneficial effects of this invention are reflected in the synergistic mechanism at three levels: prescription, drug delivery system, and process sequence. Specifically, at the prescription level, the full-spectrum extracts of Centella asiatica, Salvia miltiorrhiza, Panax notoginseng, Eucommia ulmoides, Glycyrrhiza uralensis, Boswellia carterii, Curcuma longa, and Ligusticum chuanxiong complement each other in inflammation regulation, anti-oxidation, microcirculation improvement, and tissue remodeling: polyphenols and flavonoids inhibit inflammatory pathways such as NF-κB, COX-2, and MAPK and scavenge reactive oxygen species; triterpenes and steroidal saponins reduce inflammatory mediators and exudation; lactones and aromatic components improve hemorheology and endothelial nitric oxide signaling; and triterpenoid glycosides and lignan-like components regulate fibroblast activity, MMP / TIMP, and TGF-β / Smad axes to optimize collagen deposition and inhibit adhesion. At the same time, licorice-derived components improve skin tolerance and buffer potential irritation, enabling the three effect chains of analgesia, anti-inflammation, and repair promotion to advance continuously in the same local space and time. At the drug delivery system level, the hydrophilic extract enhances solubility and chemical stability through cyclodextrin inclusion and reduces the risk of irritation associated with highly polar cosolvents. The lipophilic extract exists in the form of water-in-oil nanoemulsions, improving the partition coefficient of the stratum corneum and its translayer diffusion ability. Surfactants and phospholipids work together to stabilize the oil-water interface, reducing particle aggregation and uneven migration. The two drug repositories operate in parallel within the same system, allowing the intradermal concentration-time curves of components with different physicochemical properties to be complementary, thereby simultaneously covering the onset and maintenance phases. The thermosensitive gel matrix gels in situ under skin temperature conditions, forming a drug delivery chamber with local retention and slow diffusion. This reduces sagging and accidental migration and provides a continuous aqueous network for nanoemulsion particles and inclusion complexes, achieving a synergistic improvement in steady-state flux and intra-tissue retention through a diffusion-penetration-release coupling mechanism. Two molecular weight hyaluronic acid serves as both film-forming lubricant and deep moisturizer, reducing local friction and shear, improving the microenvironment of tendon sheaths and synovium, and facilitating the controlled diffusion of components at the epidermal-dermal interface. At the process level, the cold-swelling thermosensitive matrix prevents premature gelation; encapsulating the hydrophilic extract before batching with the matrix reduces free precipitation and local concentration gradients; the lipophilic extract undergoes high-shear and ultrasonic treatment to form a narrow-distribution nanoemulsion before incorporation, reducing phase separation caused by interfacial recombination; low-temperature batch integration and vacuum degassing ensure particle size distribution and microscopic uniformity; and a low-ionic-strength buffer system maintains the compatibility pH and reduces the interference of electrolytes on rheology and particle size. The combination of these sequences and conditions ensures the stability of the formulation-carrier-structure at the microscopic level. Furthermore, this formulation is a topical thermosensitive gel, which can gel in situ at skin temperature to form a local drug reservoir. The embedded O / W nanoemulsion within the system achieves stable delivery of the lipophilic components. In summary, the pharmacological complementarity of the eight full-spectrum extracts and the synergistic effect of the biphasic drug loading-thermosensitive reservoir-rheological optimization system design achieve simultaneous anti-inflammatory, analgesic, anti-swelling, and functional repair mechanisms. The final product demonstrates significant advantages in inflammation suppression, pain relief, edema reduction, and orderly collagen remodeling, while also exhibiting superior safety and skin compatibility. Attached Figure Description

[0006] Figure 1 This is a graph showing the biocompatibility test results of the traditional Chinese medicine compositions of Examples 1-3 and Comparative Examples 1-4 of the present invention; Figure 2 This is a graph showing the in vitro release hydrophilicity index test results of the traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 of the present invention; Figure 3 This is a graph showing the in vitro release lipophilic index test results of the traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 of the present invention; Figure 4 This is a graph showing the viscosity change results of the stability test of the traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 of the present invention; Figure 5 This is a graph showing the pH change results of the stability test of the traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 of the present invention; Figure 6 The graph shows the anti-inflammatory test results of the traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 of this invention. Detailed Implementation

[0007] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Unless otherwise specified, the raw materials involved in the present invention are all purchased through conventional commercial channels. Experimental methods without specific conditions are conventional methods and conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.

[0008] Example 1: A traditional Chinese medicine composition for treating orthopedic soft tissue injuries, comprising the following raw materials in parts by weight: 5 parts Centella asiatica extract, 5 parts Salvia miltiorrhiza extract, 3 parts Panax notoginseng extract, 3 parts Eucommia ulmoides extract, 3.5 parts Glycyrrhiza uralensis extract, 6 parts Boswellia carterii resin extract, 6 parts Curcuma longa extract and 4 parts Ligusticum chuanxiong extract. The traditional Chinese medicine composition for treating orthopedic soft tissue injuries further comprises the following raw materials in parts by weight: 200 parts poloxamer 407, 25 parts poloxamer 188, 1.5 parts high molecular weight hyaluronic acid, 0.6 parts low molecular weight hyaluronic acid, 24 parts hydroxypropyl-β-cyclodextrin, 0.5 parts penetration enhancer, 50 parts oil phase, 10 parts polyoxyethylene hydrogenated castor oil, 40 parts Tween-80, 18 parts propylene glycol, 3.5 parts phenoxyethanol, 12 parts ethanol, and 8 parts lecithin; The preparation of the traditional Chinese medicine composition for treating orthopedic soft tissue injuries specifically includes the following steps: S1: Slowly add poloxamer 407 and poloxamer 188 to deionized water at 4°C, stir to dissolve evenly, and let stand at 4°C for 10 hours to obtain phase A; S2: Slowly add high molecular weight hyaluronic acid and low molecular weight hyaluronic acid to deionized water at 4°C, stir at 250 rpm for 50 min to hydrate evenly, then add part of propylene glycol, stir to dissolve evenly, and obtain phase C; S3: Add hydroxypropyl-β-cyclodextrin to deionized water, stir at 500 rpm for 12 min, then slowly add Centella asiatica extract, Salvia miltiorrhiza extract, Panax notoginseng extract, Eucommia ulmoides extract and Glycyrrhiza uralensis extract in sequence, stirring for 10 min after each addition; then filter at 0.45 μm to obtain phase B; S4: Under 60℃ water bath conditions, medium-chain triglycerides and lecithin were mixed at 500 rpm for 12 min, then oleic acid was added and stirred until dissolved. After that, frankincense resin extract, turmeric extract and chuanxiong extract were dissolved in sequence, stirring for 8 min after each addition of each extract. Then Tween-80, polyoxyethylene hydrogenated castor oil and ethanol were added, and stirring was continued at 60℃ for 10 min. Then the rotation speed was increased to 10000 rpm for shearing treatment for 4 min, followed by ultrasonic treatment at 300W, 5s on / 5s off, for a total of 4 min. The temperature was controlled below 25℃ in an ice bath throughout the process to obtain phase D. S5: Dissolve phenoxyethanol in an equal weight of propylene glycol to obtain phase E; S6: Place phase A from step S1 in a container and stir continuously at 300 rpm. Then, slowly add phase C from step S2 over 12 min, maintaining the material temperature at 6°C. Add phase B from step S3 over 25 min, maintaining the material temperature at 6°C. Add phase D from step S4 in a thin stream over 8 min, increasing the speed to 500 rpm and maintaining the material temperature at 6°C. Continue stirring for 20 min after adding the phase D. Add phase E from step S5, stir for 10 min, add 10 mM PBS buffer to adjust the pH to 6.5, degas under vacuum at -0.08 MPa for 8 min, and repeat the process twice to obtain the traditional Chinese medicine composition.

[0009] Example 2: A traditional Chinese medicine composition for treating orthopedic soft tissue injuries, comprising the following raw materials in parts by weight: 6 parts Centella asiatica extract, 6 parts Salvia miltiorrhiza extract, 6 parts Panax notoginseng extract, 3.5 parts Eucommia ulmoides extract, 4 parts Glycyrrhiza uralensis extract, 4 parts Boswellia carterii resin extract, 6.5 parts Curcuma longa extract and 3 parts Ligusticum chuanxiong extract. The traditional Chinese medicine composition for treating orthopedic soft tissue injuries further comprises the following raw materials in parts by weight: 220 parts poloxamer 407, 20 parts poloxamer 188, 2 parts high molecular weight hyaluronic acid, 0.8 parts low molecular weight hyaluronic acid, 26 parts hydroxypropyl-β-cyclodextrin, 0.1 parts penetration enhancer, 40 parts oil phase, 6 parts polyoxyethylene hydrogenated castor oil, 35 parts Tween-80, 15 parts propylene glycol, 3 parts phenoxyethanol, 10 parts ethanol, and 12 parts lecithin; The preparation of the traditional Chinese medicine composition for treating orthopedic soft tissue injuries specifically includes the following steps: S1: Slowly add poloxamer 407 and poloxamer 188 to deionized water at 4°C, stir to dissolve evenly, and let stand at 4°C for 12 hours to obtain phase A; S2: Slowly add high molecular weight hyaluronic acid and low molecular weight hyaluronic acid to deionized water at 4°C, stir at 300 rpm for 60 min to hydrate evenly, then add some propylene glycol, stir to dissolve evenly, and obtain phase C; S3: Add hydroxypropyl-β-cyclodextrin to deionized water, stir at 600 rpm for 15 min, then slowly add Centella asiatica extract, Salvia miltiorrhiza extract, Panax notoginseng extract, Eucommia ulmoides extract and Glycyrrhiza uralensis extract in sequence, stirring for 10 min after each addition; then filter at 0.45 μm to obtain phase B; S4: Under 60℃ water bath conditions, medium-chain triglycerides and lecithin were mixed at 500 rpm for 15 min, then oleic acid was added and stirred until dissolved. Then, frankincense resin extract, turmeric extract and chuanxiong extract were dissolved in sequence, stirring for 10 min after each addition. Tween-80, polyoxyethylene hydrogenated castor oil and ethanol were then added, and stirring was continued at 60℃ for 10 min. Then, the rotation speed was increased to 12000 rpm for shearing treatment for 5 min, followed by ultrasonic treatment at 400W, 5s on / 5s off, for a total of 5 min. The temperature was controlled below 25℃ in an ice bath throughout the process to obtain phase D. S5: Dissolve phenoxyethanol in an equal weight of propylene glycol to obtain phase E; S6: Place phase A from step S1 in a container and stir continuously at 300 rpm. Then, slowly add phase C from step S2 over 15 min, maintaining a material temperature of 5°C. Add phase B from step S3 over 30 min, maintaining a material temperature of 5°C. Add phase D from step S4 in a thin stream over 10 min, increasing the speed to 500 rpm and maintaining a material temperature of 5°C. Continue stirring for 20 min after adding the phase D. Add phase E from step S5, stir for 10 min, add 10 mM PBS buffer to adjust the pH to 6.4, degas under vacuum at -0.08 MPa for 10 min, and repeat the process twice to obtain the traditional Chinese medicine composition.

[0010] Example 3: A traditional Chinese medicine composition for treating orthopedic soft tissue injuries, comprising the following raw materials in parts by weight: 4 parts Centella asiatica extract, 4 parts Salvia miltiorrhiza extract, 4 parts Panax notoginseng extract, 2 parts Eucommia ulmoides extract, 2 parts Glycyrrhiza uralensis extract, 7 parts Boswellia carterii resin extract, 4 parts Curcuma longa extract and 4.5 parts Ligusticum chuanxiong extract; The traditional Chinese medicine composition for treating orthopedic soft tissue injuries further comprises the following raw materials in parts by weight: 180 parts poloxamer 407, 30 parts poloxamer 188, 1 part high molecular weight hyaluronic acid, 0.3 parts low molecular weight hyaluronic acid, 18 parts hydroxypropyl-β-cyclodextrin, 1 part penetration enhancer, 60 parts oil phase, 12 parts polyoxyethylene hydrogenated castor oil, 50 parts Tween-80, 20 parts propylene glycol, 4 parts phenoxyethanol, 15 parts ethanol, and 12 parts lecithin; The preparation of the traditional Chinese medicine composition for treating orthopedic soft tissue injuries specifically includes the following steps: S1: Slowly add poloxamer 407 and poloxamer 188 to deionized water at 4°C, stir to dissolve evenly, and let stand at 4°C for 8 hours to obtain phase A; S2: Slowly add high molecular weight hyaluronic acid and low molecular weight hyaluronic acid to deionized water at 4°C, stir at 200 rpm for 30 min to hydrate evenly, then add some propylene glycol, stir to dissolve evenly, and obtain phase C; S3: Add hydroxypropyl-β-cyclodextrin to deionized water, stir at 400 rpm for 10 min, then slowly add Centella asiatica extract, Salvia miltiorrhiza extract, Panax notoginseng extract, Eucommia ulmoides extract and Glycyrrhiza uralensis extract in sequence, stirring for 10 min after each addition; then filter at 0.45 μm to obtain phase B; S4: Isopropyl palmitate and lecithin were mixed at 500 rpm for 10 min in a 60℃ water bath. The temperature was then lowered to 30℃, borneol was added, and the mixture was stirred until dissolved. Boswellia resin extract, turmeric extract, and Ligusticum chuanxiong extract were dissolved sequentially, with each extract added and stirred for 5 min. Tween-80, polyoxyethylene hydrogenated castor oil, and ethanol were then added, and the mixture was stirred at 60℃ for another 10 min. The mixture was then sheared at 8000 rpm for 3 min, followed by ultrasonic treatment at 200W for 5 s on and 5 s off, for a total of 3 min. The temperature was controlled below 25℃ throughout the process in an ice bath to obtain phase D. S5: Dissolve phenoxyethanol in an equal weight of propylene glycol to obtain phase E; S6: Place phase A from step S1 in a container and stir continuously at 300 rpm. Then, slowly add phase C from step S2 over 10 min, maintaining the material temperature at 10°C. Add phase B from step S3 over 20 min, maintaining the material temperature at 10°C. Add phase D from step S4 dropwise over 5 min in a thin stream, increasing the rotation speed to 500 rpm and maintaining the material temperature at 10°C. Continue stirring for 20 min after adding the phase D. Add phase E from step S5, stir for 10 min, add 10 mM PBS buffer to adjust the pH to 6.6, degas under vacuum at -0.08 MPa for 5 min, and repeat the process twice to obtain the traditional Chinese medicine composition.

[0011] Comparative Example 1: The operating parameters of Comparative Example 1 are basically the same as those of Example 1. The main difference is that the cyclodextrin inclusion step of the hydrophilic extract is omitted in Comparative Example 1. That is, in step S3, hydroxypropyl-β-cyclodextrin is not added, and the extracts of Centella asiatica, Salvia miltiorrhiza, Panax notoginseng, Eucommia ulmoides, and Glycyrrhiza uralensis are directly dispersed in water and then incorporated into the matrix. The remaining process parameters remain unchanged.

[0012] Comparative Example 2: The operating parameters of Comparative Example 2 are basically the same as those of Example 1. The main difference is that the construction step of the lipophilic extract nanoemulsion is omitted in Comparative Example 2. That is, high shear and ultrasonic emulsification are not performed in step S4. Instead, the frankincense resin extract, turmeric extract and chuanxiong extract are directly dissolved in the oil phase and then incorporated into the system in step S6. The remaining process parameters remain unchanged.

[0013] Comparative Example 3: The operation parameters of Comparative Example 3 are basically the same as those of Example 1. The main difference is that Comparative Example 3 does not use low-temperature cold swelling and low-temperature batching in step S1 and batching-related step S6, but performs swelling and phase integration at room temperature. The remaining process parameters remain unchanged.

[0014] Comparative Example 4: The operating parameters of Comparative Example 4 are basically the same as those of Example 1. The main difference is that the Ligusticum chuanxiong extract in the eight-ingredient composition is removed in Comparative Example 4, while the original ratio of the other seven ingredients is maintained and the total amount is made up with deionized water. The other process parameters remain unchanged.

[0015] Performance testing: Biocompatibility testing: The traditional Chinese medicine compositions from Examples 1-3 and Comparative Examples 1-4 were selected. Cell compatibility was evaluated on human dermal fibroblasts (HDF) using a culture medium extraction method. Each sample was added to DMEM medium containing 10% FBS at 0.2 g / mL (sample mass / culture medium volume) under aseptic conditions. After gentle shaking and extraction at 37°C for 24 h, the supernatant was collected by centrifugation and sterilized through a 0.22 μm filter to obtain the 100% extract. Simultaneously, a blank matrix extract (consistent with the formulation of the examples but without the traditional Chinese medicine extract) was prepared to differentiate the influence of the carrier. HDF cells were seeded into 96-well plates and cultured to 70-80% confluence before discarding the substrate. 100 μL of each group's extract was added to each well, and the plates were incubated at 37°C and 5% CO2 for 24 h and 48 h, respectively. The negative control was fresh culture medium, and the positive control was 10% dimethyl sulfoxide medium. After incubation, CCK-8 assays were performed according to the reagent instructions to determine OD. 450 The survival rate (%) was calculated as follows: (OD sample - OD positive control) / (OD negative control - OD positive control) × 100%. The results are as follows: Figure 1 As shown.

[0016] based on Figure 1The results showed that the traditional Chinese medicine composition prepared in this invention maintained a high cell viability at both time points, demonstrating excellent biocompatibility. In Comparative Example 1, after removing cyclodextrin, the hydrophilic extract existed in a free or micro-aggregated form, making it more prone to forming local concentration peaks and osmotic pressure shifts in the early stages of extraction. Simultaneously, due to the lack of inclusion shielding, some polyphenols / saponins interacted non-specifically with the carrier matrix, resulting in an increase in transient load on the cell membrane surface and a decrease in metabolic activity, manifesting as a lower cell viability than in Example 1. In Comparative Example 2, no nanoemulsion was constructed; the lipophilic extract was dissolved only in the oil phase and directly incorporated into the system. The oil droplet size distribution and interfacial stability were insufficient. During early extraction, the partition coefficient of the lipophilic component in the culture medium increased, and the local microenvironment surface tension changed more significantly, resulting in more primitive hydrophobic micro-clusters encountered by the cells. Although the overall stimulation was not strong, the biocompatibility was lower than in Example 1. In Comparative Example 3, after eliminating low-temperature swelling and low-temperature batching, the probability of premature local gelation and reorganization of the matrix during batch integration increased. The spatial distribution of hydrophilic inclusion complexes and lipophilic oil droplets in the gel network became uneven, accompanied by particle size drift and a small amount of microbubbles / microphase separation. During extraction, uneven release of soluble components and surfactants and slight pH / osmotic pressure fluctuations were more likely to occur, resulting in poorer stability of the interfacial environment of cells and a relatively consistent decline in survival rate. In Comparative Example 4, after removing Ligusticum chuanxiong, the volatile and aromatic lipophilic components in the system decreased, and the potential transient olfactory / irritant sources were reduced. Under the premise of maintaining the established carrier and process, the interfacial properties and osmotic pressure of the extraction phase remained basically unchanged. Therefore, the overall compatibility was close to that of Example 1, which is consistent with our previous understanding that the synergistic effect of the eight ingredients mainly serves the closed-loop effect rather than determining compatibility.

[0017] In vitro release test: The traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 were selected. The drug release capacity was evaluated using a Franz diffusion cell combined with an inert synthetic membrane. Cellulose acetate (pore size 0.45 μm) was equilibrated in the recipient solution (10 mM salt-free PBS, pH 6.5, containing 0.5% Tween-80 and 2% propylene glycol to maintain the leak conditions) for 60 min and then clamped. The recipient chamber was kept at a constant temperature of 32°C and magnetically stirred. The donor chamber was uniformly coated with a limited dose of sample (5 mg / cm³). 2 Receptor fluid samples were collected at 0.5, 1, 2, 4, 6, 8, and 12 hours, and replenished with an equal volume of fresh receptor fluid. A negative control was a matrix blank (containing no herbal extracts). Samples were filtered through a 0.22 μm membrane and then simultaneously quantified using validated HPLC / UPLC to determine representative hydrophilic (tanshinone B, derived from tanshinone extract) and lipophilic (AKBA, derived from frankincense resin extract) components. The cumulative release per unit area, Q, was calculated. (t) (ug / cm 2 The result is as follows Figure 2 and Figure 3 As shown.

[0018] based on Figure 2 and Figure 3 Analysis of the results showed that the two representative curves of the traditional Chinese medicine composition prepared in the embodiments of the present invention (hydrophilic end of salvianolic acid B and lipophilic end of AKBA) exhibited the characteristics of sufficient release drive in the initial stage, controlled diffusion in the subsequent stage, and overall smoothness. In Comparative Example 1, the removal of cyclodextrin inclusion resulted in a decrease in the apparent saturated solubility and poorer free-state dispersibility of the hydrophilic end, weakening the effective concentration gradient at the membrane surface, leading to a significant decrease in the early release constant and cumulative release. The lipophilic end was independent of cyclodextrin, therefore its impact on the AKBA curve was limited, mainly manifested as an overall morphology similar to the examples but with slight fluctuations. In Comparative Example 2, after the nanoemulsion construction was removed, the lipophilic end transformed into a state of coarse oil droplets / dissolved in bulk oil, significantly reducing the oil droplet-water phase interface area and increasing the interfacial tension. Transmembrane partitioning and acceptor phase leakage conditions were both suppressed, resulting in a limited AKBA release rate and cumulative amount throughout the process. The hydrophilic end still had cyclodextrin inclusion and aqueous diffusion channels, and the curve of salvianolic acid B only decreased slightly. In Comparative Example 3, after abandoning low-temperature swelling and low-temperature batch integration, localized premature gelation and reorganization occurred during the batch integration process. The spatial distribution of inclusion complexes and oil droplets in the gel network was uneven, accompanied by an increased risk of particle size drift and microphase separation. The effective diffusion area and pathway connectivity of the membrane surface were weakened, resulting in a systematic decrease in both the early release and steady-state phases at the hydrophilic and lipophilic ends. The curves became more blunt, the hysteresis time was prolonged, and the intra-batch dispersion increased. In Comparative Example 4, while maintaining the cyclodextrin inclusion, nanoemulsion structure, and low-temperature sequence, only the Ligusticum chuanxiong extract was removed. The hydrophilic and lipophilic channels, interfacial structure, and gel diffusion network of the system remained unchanged. Therefore, the release curves of salvianolic acid B and AKBA were closer to those of the examples. The slight differences at the lipophilic end may mainly come from subtle changes in the overall oil phase and volatile component composition of the formulation, without affecting the transmembrane distribution mechanism dominated by the nanoemulsion.

[0019] Stability test: The traditional Chinese medicine compositions of Example 1 and Comparative Examples 1-4 were selected, sealed under nitrogen, and placed in an environment of 40°C and 75%RH for 6 months. Samples were taken at months 1, 2, 3, and 6 to test their viscosity and pH changes. The results are as follows: Figure 4 and Figure 5 As shown.

[0020] based on Figure 4 and Figure 5Analysis of the results showed that the viscosity and pH curves of the traditional Chinese medicine compositions prepared in the embodiments of the present invention exhibited a smooth and stable characteristic with minimal fluctuations, demonstrating excellent overall stability. In Comparative Example 1, the lack of inclusion complexation in the decyclodextrin resulted in the hydrophilic extract existing in a free or micro-aggregated form, which was more prone to self-aggregation / precipitation over time and to hydrogen bonding and hydrophobic association with the poloxamer / hyaluronic acid network (PF / HA network), disrupting local network uniformity and leading to a greater decrease in viscosity. Simultaneously, the solubility and chemical stability of polyphenols and saponins decreased, making them more susceptible to oxidation or hydrolysis, producing weakly acidic byproducts. This, combined with the osmotic pressure micro-drift during the extraction period, resulted in a more significant pH decline. In Comparative Example 2, after removing the nanoemulsion, the lipophilic component was only soluble in the bulk oil, existing as coarse oil droplets. The oil-water interface area decreased and the interfacial tension increased. Over time, droplet formation and slow phase separation occurred, promoting the redistribution of surfactants and the oil phase in the gel network, disrupting local microscopic homogeneity, resulting in a moderate decrease in viscosity. Due to the protection of cyclodextrin at the hydrophilic end, the accumulation of acidic byproducts was limited, and the pH change was smaller and closer to that of the Example. In Comparative Example 3, after removing the low-temperature sequence, the room-temperature swelling and the easily triggered local premature gelation of the system were inconsistent with the shear history. The spatial distribution of hydrophilic inclusion complexes and lipophilic oil droplets in the network showed initial non-uniformity. Under accelerated conditions, this initial defect was amplified, manifested as more significant oil droplet reforming, microphase separation, and bubble residue. The network connectivity deteriorated, and the viscosity decreased continuously and significantly over time. At the same time, the non-uniform network increased the contact probability between free components and the interface, slightly promoting oxidation / hydrolysis, resulting in a significant change in pH compared to the Example. Comparative Example 4, without Ligusticum chuanxiong, maintained the same carrier and process, only reducing some volatile / aromatic lipophilic components. This had a limited impact on the core stabilization mechanisms of the system network, cyclodextrin inclusion, and nanoemulsion structure. The time trajectories of viscosity and pH were similar to those of the examples.

[0021] In vitro anti-inflammatory test: Examples 1, Comparative Examples 1-4, and matrix blank (consistent with the prescription but without Chinese herbal extracts) were selected as test samples. Under aseptic conditions, they were added to DMEM medium containing 10% FBS at a ratio of 0.2 g / mL (sample mass / culture medium volume), and gently shaken at 37°C for 24 h. The supernatant was collected by centrifugation and sterilized through a 0.22 μm filter. The obtained supernatant was 100% extract. Fresh culture medium (negative control) and 1 μM dexamethasone solution (positive control) were prepared simultaneously. RAW264.7 cells were seeded in 96-well plates (1×10⁻⁶ cells / wells). 5Incubate at 37°C and 5% CO2 overnight (100 μL / well) until stable adhesion, then discard the substrate. Add 100 μL of 100% extract of each group to each well for pre-incubation for 2 h, then add LPS to a final concentration of 100 ng / mL and continue incubation for 24 h. Negative control wells do not contain LPS, only fresh culture medium. Inflammation control is (LPS + matrix blank extract). Positive control is (LPS + dexamethasone). After incubation, collect the supernatant and perform TNF-α ELISA according to the kit instructions. TNF-α inhibition rate (%) = [mean TNF-α inflammation control - mean TNF-α treatment group] / mean TNF-α inflammation control × 100%. Results are shown below. Figure 6 As shown.

[0022] based on Figure 6 Results analysis showed that Example 1 of the present invention exhibited a high anti-inflammatory inhibition rate, which was significantly better than the control. Although the effect was significantly weaker than the positive control, the inhibition intensity of Example 1 was within a reasonable range that was meaningful but lower than that of glucocorticoids. Dexamethasone directly intervenes in the expression of a broad spectrum of inflammatory genes through receptor-mediated transcriptional inhibition, and the inhibition amplitude is usually higher in acute in vitro models. In contrast, the present invention relies on the joint regulation and exposure optimization of nodes such as NF-κB / COX-2 by multiple components. Its advantages lie in multi-channel delivery and mild compatibility. The anti-inflammatory effect can be synergistic with pathways such as microcirculation improvement and tissue repair, and is not characterized by a single strong immunosuppression. In Comparative Example 1, the removal of cyclodextrin reduced the apparent solubility and diffusion driving force of hydrophilic polyphenols, making them more prone to non-specific interactions with the matrix or proteins, resulting in insufficient effective concentrations near the action sites and a significant weakening of TNF-α inhibition compared to Example 1. In Comparative Example 2, no nanoemulsion was constructed, and the lipophilic activity existed only in the form of coarse oil droplets or bulk oil, significantly reducing the interfacial area and transmembrane partition coefficient, resulting in insufficient extracellular effective exposure and further reducing TNF-α inhibition. In Comparative Example 3, abandoning the low-temperature sequence amplified the initial heterogeneity of the gel network and the drift of oil droplet size, leading to a discrete spatiotemporal distribution of effective components, unstable signal pathway regulation, and a systematic deterioration of the overall anti-inflammatory effect. In Comparative Example 4, with the carrier and process kept consistent, the removal of Ligusticum striatum retained both hydrophilic and lipophilic delivery channels, but lost the synergistic effect of some aromatic components on NF-κB-related nodes, thus showing a moderate decrease in inhibitory ability compared to Example 1.

[0023] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A traditional Chinese medicine composition for treating orthopedic soft tissue injury, characterized in that, The composition comprises the following raw materials by weight parts: 4-6 parts of Centella asiatica extract, 4-6 parts of Salvia miltiorrhiza extract, 4-6 parts of Panax notoginseng extract, 2-3.5 parts of Eucommia ulmoides extract, 2-4 parts of Glycyrrhiza extract, 4-7 parts of Boswellia resin extract, 4-6.5 parts of Curcuma longa extract, and 3-4.5 parts of Ligusticum chuanxiong extract.

2. The traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 1, characterized in that, The composition of the traditional Chinese medicine composition comprises the following raw materials by weight parts: 5 parts of Centella asiatica extract, 5 parts of Salvia miltiorrhiza extract, 3 parts of Panax notoginseng extract, 3 parts of Eucommia ulmoides extract, 3.5 parts of Glycyrrhiza extract, 6 parts of Boswellia resin extract, 6 parts of Curcuma longa extract, and 4 parts of Ligusticum chuanxiong extract.

3. The traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 2, characterized in that, The traditional Chinese medicine composition for treating orthopedic soft tissue injury further comprises the following raw materials by weight parts: 180-220 parts of poloxamer 407, 20-30 parts of poloxamer 188, 1-2 parts of high molecular hyaluronic acid, 0.3-0.8 parts of low molecular hyaluronic acid, 18-26 parts of hydroxypropyl-β-cyclodextrin, 0.1-1 parts of penetration enhancer, 40-60 parts of oil phase, 6-12 parts of polyoxyethylene hydrogenated castor oil, 35-50 parts of Tween-80, 15-20 parts of propylene glycol, 3-4 parts of phenoxy ethanol, 10-15 parts of ethanol, and 5-12 parts of lecithin.

4. The traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 3, characterized in that, The low molecular weight hyaluronic acid has a molecular weight of 30-100 kDa; and the high molecular weight hyaluronic acid has a molecular weight of 800-1200 kDa.

5. The traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 4, characterized in that, The penetration enhancer is any one of oleic acid, menthol, and borneol.

6. The traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 5, characterized in that, The oil phase is any one of medium-chain triglyceride and isopropyl palmitate.

7. A method for preparing the traditional Chinese medicine composition for treating orthopedic soft tissue injury according to any one of claims 1-6, characterized in that, Specifically comprising the following steps: S1: slowly add poloxamer 407 and poloxamer 188 into deionized water, stir to dissolve uniformly, stand, and obtain phase A; S2: slowly add high molecular weight hyaluronic acid and low molecular weight hyaluronic acid into deionized water, stir to hydrate uniformly, then add part of propylene glycol, stir to dissolve uniformly, and obtain phase C; S3: add hydroxypropyl-β-cyclodextrin into deionized water, stir, then slowly add Centella asiatica extract, Salvia miltiorrhiza extract, Panax notoginseng extract, Eucommia ulmoides extract, and Glycyrrhiza extract in sequence, each time after stirring after adding one kind of extract, then add the subsequent extract; filter, and obtain phase B; S4: under the condition of heating water bath, mix the oil phase and lecithin by stirring, then add the penetration enhancer, stir to dissolve uniformly, then dissolve Boswellia resin extract, Curcuma longa extract, and Ligusticum chuanxiong extract in sequence, each time after stirring after adding one kind of extract; then add Tween-80, polyoxyethylene hydrogenated castor oil, and ethanol, continue to stir, then improve the rotation speed for shearing treatment, and ultrasonic treatment, and obtain phase D; S5: dissolve phenoxy ethanol in equal weight parts of propylene glycol, and obtain phase E; S6: place phase A of step S1 in a container, continuously stir, then slowly add phase C of step S2, maintain low material temperature; add phase B of step S3, maintain low material temperature; add phase D of step S4 in a thin stream mode, improve the rotation speed, maintain low material temperature, and continue to stir after being incorporated; add phase E of step S5, stir, add PBS buffer solution for pH adjustment, and vacuum degassing, and obtain the traditional Chinese medicine composition.

8. A method for preparing the traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 7, characterized in that, In step S4, the ultrasonic treatment parameters are: 200-400 W, 5 s on / 5 s off, cumulative 3-5 min, and the whole process is controlled at a temperature lower than 25℃ by ice bath.

9. A method for preparing the traditional Chinese medicine composition for treating orthopedic soft tissue injury according to claim 8, characterized in that, In step S6, the vacuum defoaming parameters are: -0.08 MPa, time 5-10 min, and repeated treatment 2 times.

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