Composition for improving bone and joint health and application thereof

Abstract

The present invention discloses a composition for improving bone and joint health and its application, comprising glucosamine and turmeric powder; in terms of mass ratio, the mass ratio of the turmeric powder to glucosamine is 1:(0.2-5). The present invention proposes a composition for regulating inflammatory factors and improving subchondral bone damage, with glucosamine and turmeric powder as main functional ingredients, wherein the mass ratio of glucosamine to turmeric powder is 3:1, and the obtained composition is verified by cell tests and animal tests to have the efficacy of regulating the level of inflammatory factors and improving subchondral bone damage.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional compositions, and in particular relates to a composition for improving bone and joint health and application thereof. Background Art

[0002] Osteoarthritis (OA), the most common form of arthritis, is a synovial joint disease characterized by cartilage degradation and bony overgrowth manifested as osteophytes and subchondral thickening. OA is also associated with varying degrees of synovitis and damage to other joint structures, including the ligaments and menisci and subchondral bone of the knee.

[0003] Turmeric is a rhizome plant of the ginger family. It has become the first choice of alternative medicine due to its anti-inflammatory, antioxidant and digestive functions. Its main ingredient, curcumin, is also a natural reactive oxygen scavenger. It has been shown to have an effective therapeutic effect on pain caused by arthritis and OA. Its main mechanism may be related to protecting IL-1b-induced apoptotic chondrocytes, improving early degenerative changes in articular cartilage, and inhibiting the production of cytoplasmic phospholipase A2 (cPLA2), cyclooxygenase 2 (COX-2), 5-lipoxygenase (5-LOX), etc.

[0004] Glucosamine is a precursor for the synthesis of glycosylated proteins and lipids. This natural compound is one of the most abundant monosaccharides in the human body and has an oral bioavailability of 20%. In vitro studies have shown that the addition of glucosamine to chondrocyte cultures can increase the synthesis of aggregated proteins and reduce the inflammatory process by reducing the catabolic activity of proteases; the positive effects of glucosamine may be due to its anti-inflammatory activity, stimulation of proteoglycan synthesis, and inhibition of proteolytic enzyme synthesis. It is also believed that glucosamine promotes the production of synovial hyaluronic acid and inhibits cartilage activity by degrading liposome enzymes.

[0005] Subchondral bone usually refers to the epiphyseal bone area located just below the calcified cartilage. From an anatomical perspective, the subchondral bone structure can be further divided into subchondral bone plates and subchondral bone trabeculae. Subchondral bone sclerosis and progressive cartilage degeneration are one of the hallmarks of osteoarthritis. Studies have shown that different microstructural changes in subchondral bone occur at different stages of osteoarthritis. In the early stages of osteoarthritis, bone remodeling and enhanced subchondral bone absorption are considered to be the determining factors for its progression. In the late stages of osteoarthritis, subchondral bone shows reduced bone absorption and bone turnover, while bone formation is relatively increased. The method of targeting subchondral bone for treatment has shown good efficacy in animal experiments.

[0006] Currently, the existing glucosamine and turmeric compositions have not found the regulatory effects of glucosamine or turmeric alone / in combination on subchondral bone damage. Summary of the invention

[0007] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0008] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0009] Therefore, the object of the present invention is to overcome the deficiencies in the prior art and provide a composition for regulating inflammatory factors and improving subchondral bone damage.

[0010] In order to solve the above technical problems, the present invention provides the following technical solution: a composition for regulating inflammatory factors and improving subchondral bone damage, comprising glucosamine and turmeric powder.

[0011] As a preferred embodiment of the composition of the present invention, the mass ratio of the turmeric powder to the glucosamine is 1:(0.2-5) by mass ratio.

[0012] As a preferred embodiment of the composition of the present invention, the mass ratio of the turmeric powder to the glucosamine is 1:(0.3-3).

[0013] As a preferred embodiment of the composition of the present invention, the curcumin content in the turmeric powder is ≥1wt%.

[0014] The composition of the present invention can improve bone and joint health by regulating inflammation and improving subchondral bone damage.

[0015] As a preferred embodiment of the composition of the present invention, the inflammatory factors include one or more of NO, MMP13, TNF-α and IL-6.

[0016] Another object of the present invention is to overcome the deficiencies in the prior art and provide a composition for use in preparing foods and medicines that have the efficacy of regulating inflammatory factors and improving subchondral bone damage.

[0017] As a preferred embodiment of the application of the present invention, the inflammatory factors include one or more of NO, MMP13, TNF-α and IL-6.

[0018] Another object of the present invention is to overcome the deficiencies in the prior art and provide a food comprising a composition for regulating inflammatory factors and improving subchondral bone damage and a food science-acceptable auxiliary material.

[0019] As a preferred embodiment of the food of the present invention, the dosage form of the food includes tablets, capsules, drinks, soft candies or powders.

[0020] Beneficial effects of the present invention:

[0021] (1) The present invention provides a composition for regulating inflammatory factors and improving subchondral bone damage, with glucosamine and turmeric powder as main effective ingredients, wherein the mass ratio of glucosamine to turmeric powder is 3:1. The prepared composition is verified by cell tests and animal tests to have the efficacy of regulating the level of inflammatory factors and improving subchondral bone damage.

[0022] (2) The composition of the present invention can be used as a main ingredient in foods that relieve joint discomfort by regulating the level of inflammatory factors and improving subchondral bone damage. It can be made into capsules, compressed candies, etc., and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:

[0024] Figure 1 1 is a flow chart of cell experiments in an embodiment of the present invention.

[0025] Figure 2 The figure is a graph showing the effect of different ratios of glucosamine and turmeric on LPS-induced cell inflammation in the embodiment of the present invention, wherein: b vs model group, p<0.05; c vs glucosamine group, p<0.05; d vs turmeric group, p < 0.05; e vs 1:1 group, p<0.05; f vs1:3 groups, p<0.05; g vs3:1 group, p<0.05.

[0026] Figure 3 Graph showing the improving effects of curcumin and glucosamine on subchondral bone plate damage in osteoarthritis rats in an example of the present invention.

[0027] Figure 4 Graph showing the protective effects of curcumin and glucosamine on subchondral trabecular bone in rats with osteoarthritis in an embodiment of the present invention. DETAILED DESCRIPTION

[0028] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0029] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0030] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0031] The raw materials in the embodiments of the present invention are all common commercially available products.

[0032] Example 1

[0033] This embodiment provides a method for preparing a composition for regulating inflammatory factors and improving subchondral bone damage, the main steps of which are:

[0034] (1) Prepare materials: accurately weigh 37.2 g of glucosamine hydrochloride (containing 30 g of glucosamine) and 10 g of turmeric powder (curcumin content greater than or equal to 1%);

[0035] (2) Mixing materials: 24.8 g of glucosamine hydrochloride (containing 20 g of glucosamine) and 10 g of turmeric powder are uniformly mixed and ground to prepare a composition.

[0036] Example 2

[0037] This embodiment provides a method for preparing a composition for regulating inflammatory factors and improving subchondral bone damage, the main steps of which are:

[0038] (1) Prepare materials: accurately weigh 24.8 g of glucosamine hydrochloride (containing 20 g of glucosamine) and 20 g of turmeric powder (wherein the curcumin content is greater than or equal to 1%);

[0039] (2) Mixing materials: 24.8 g of glucosamine hydrochloride (containing 20 g of glucosamine) and 20 g of turmeric powder are uniformly mixed and ground to prepare a composition.

[0040] Example 3

[0041] This embodiment provides a method for preparing a composition for regulating inflammatory factors and improving subchondral bone damage, the main steps of which are:

[0042] (1) Prepare materials: accurately weigh 12.4 g of glucosamine hydrochloride (containing 10 g of glucosamine) and 30 g of turmeric powder (wherein the curcumin content is greater than or equal to 1%);

[0043] (2) Mixing materials: 12.4 g of glucosamine hydrochloride (containing 10 g of glucosamine) and 30 g of turmeric powder are uniformly mixed and ground to prepare a composition.

[0044] Example 4

[0045] Cell assay protocol:

[0046] 1. Experimental purpose: To verify the efficacy of single glucosamine, turmeric powder and mixed samples in different proportions on LPS-induced SW1353 chondrosarcoma cell inflammation model.

[0047] 2. Experimental method: LPS-induced osteoarthritis model: the control group was added with ordinary culture medium, the model group was added with culture medium containing 5 μg / L lipopolysaccharide (LPS), and the experimental group was added with LPS + samples of different proportions grouped according to 3.1, and cultured for 24 hours.

[0048] 3. Experimental Materials and Instruments

[0049] 3.1 Experimental groups: blank group, model group, glucosamine group, turmeric powder group, glucosamine: turmeric powder 1:1 group, glucosamine: turmeric powder 1:3 group, glucosamine: turmeric powder 3:1 group;

[0050] 3.2 Experimental concentration:

[0051] Glucosamine: 500ug / ml;

[0052] Turmeric powder 500ug / ml;

[0053] Glucosamine: turmeric powder 250ug / ml: 250ug / ml;

[0054] Glucosamine: turmeric powder 125ug / ml: 375ug / ml;

[0055] Glucosamine: turmeric powder 375ug / ml: 125ug / ml.

[0056] 3.3 Experimental instruments: cell culture incubator, microplate reader, constant temperature drying oven, etc.

[0057] The experimental flow chart is shown in Figure 1 ;

[0058] The levels of inflammatory factors and MMP-13 in the cell supernatant were detected by ELISA kits. The results are shown in Figure 2It was found that compared with the normal group / control group, the model group (LPS-induced group) showed a trend of significant increase in MMP-13 and inflammatory factors, indicating that the model was successfully constructed; compared with the model group, the experimental group was able to significantly reduce the levels of inflammatory factors and MMP-13, and the effect was optimal when the ratio of glucosamine: turmeric = 3:1.

[0059] Example 5

[0060] Animal Experiment Protocol:

[0061] 1. Animals: Lewis rats, male, 9-10 weeks old, weighing about 200 grams. Ten rats per group, a total of 50 rats.

[0062] 2. Drug preparation: All drugs are dissolved or suspended in sterile water.

[0063] Group 1: control group (Sham), intragastric administration of sterile water;

[0064] Group 2: MMT modeling group (MMT), intragastric administration of sterile water;

[0065] Group 3: low-dose turmeric powder group (CL), turmeric powder was dissolved in sterile water, and the oral dose was 115 mg / kg / d;

[0066] Group 4: Glucosamine hydrochloride group (G), glucosamine hydrochloride (glucosamine content 80.62%) was dissolved in sterile water to prepare 186 mg / kg / d (glucosamine 150 mg / kg / d);

[0067] Group 5: turmeric powder + glucosamine hydrochloride group (CLG), turmeric powder 50 mg / kg / d + glucosamine hydrochloride 186 mg / kg / d (glucosamine 150 mg / kg) [glucosamine: turmeric powder = 3:1].

[0068] 4. Main steps of the experiment:

[0069] 4.1. Establishment of MMT osteoarthritis model

[0070] The animal was placed in a supine position, and the skin was longitudinally incised at the distal end of the patella and the proximal end of the tibial plateau on the medial side of the right knee. The muscle fascia was bluntly separated with ophthalmic scissors to expose the medial collateral ligament. After cutting the junction between the medial collateral ligament and the medial meniscus, the calf was moved to free the medial end of the medial meniscus. The free end was grasped with forceps, the meniscus was removed, and the muscle layer and fascia were sutured with absorbable surgical sutures, and the skin was sutured with non-absorbable sutures.

[0071] The animals in the sham group (the sham group is a sham operation group without meniscectomy) only need to follow this procedure to expose the medial collateral ligament.

[0072] 4.2 All groups of animals were given the drug by gavage on the first day after surgery, once a day, for 8 consecutive weeks.

[0073] 4.3 Detection:

[0074] Tissue sampling: Animals were euthanized after 8 weeks of treatment. The right knee joints of rats were fixed in 4% (w / v) paraformaldehyde buffer (n=10) for 3 days and then transferred to 70% alcohol. Tissue specimens were used for micro-computed tomography (Micro-CT) and histological analysis.

[0075] The right knee joints of each group were scanned by Micro CT with a resolution of 9 μm.

[0076] The subchondral bone plate and subchondral trabeculae within 2.0 mm from the sagittal highest point of the medial tibial plateau were selected as the research area.

[0077] Each subchondral bone plate and subchondral bone trabecula are reconstructed in three dimensions by post-processing multiple two-dimensional scanned images through a computer in combination with image preprocessing, algorithm model analysis and other steps to generate a continuous three-dimensional shape.

[0078] The analysis related indicators are shown in Table 1.

[0079] Table 1

[0080]

[0081]

[0082] 5. Test results:

[0083] 5.1 Curcumin and glucosamine reduce subchondral bone plate damage in OA rats

[0084] Results Figure 3 , where A is a schematic diagram for selecting the calculation / evaluation area, and the subchondral bone plate within 2.0 mm from the highest point of the sagittal plane of the medial tibial platform is selected as the region of interest; Figure B is a case image of micro CT (one image is selected from each group for illustration), and the scale line is 2 mm; Figure C is a bar graph of different indicators calculated. The analysis of the subchondral bone plate showed that the tissue volume (TV) of the MMT group (1.337±0.057) was significantly increased by 14.3% (p<0.05) compared with the sham operation group (1.230±0.045), while the tissue volume of the CLG group (1.011±0.020) was significantly reduced by 28.1% (p<0.0001) compared with the sham operation group;

[0085] Bone mineral content (BMC) was significantly decreased by 25.8% in the CLG group (0.678±0.015) compared with the MMT group (0.914±0.030) (p<0.0001).

[0086] The thickness of the subchondral bone plate in the CLG group (0.192±0.003) was 17.3% lower than that in the MMT group (0.232±0.008) (p<0.0001). The total amount of pore space (Po.V(tot)) of the subchondral bone plate in the MMT group (0.133±0.007) increased by 127.9% compared with the sham group (0.058±0.009) (p<0.0001), while that in the CLG group (0.103±0.006) decreased by 22.9% compared with the MMT group (p<0.05).

[0087] 5.2 Curcumin and glucosamine protect subchondral trabecular bone in OA rats

[0088] Results Figure 4 , where A is a schematic diagram of the selected calculation / evaluation area, and the subchondral trabeculae within 2.0 mm from the highest point of the sagittal plane of the medial tibial platform are selected as the region of interest, B is a case image of micro CT (one image is selected from each group for illustration), and the scale line is 2 mm; C is a bar graph of different indicators calculated. Analysis of subchondral trabeculae showed that the combined administration of curcumin and glucosamine had a protective effect on subchondral trabeculae in OA rats.

[0089] Subchondral trabecular bone analysis showed that the bone volume (BV) of the MMT group (2.439±0.059) was significantly reduced by 15.4% (p<0.001) compared with the sham group (2.884±0.037), while the bone volume of the C group (2.721±0.124), the G group (3.179±0.084), and the CLG group (3.136±0.074) was significantly increased by 11.6% (p<0.05), 30.4% (p<0.0001), and 28.6% (p<0.0001) respectively compared with the MMT group. The bone mineral content (BMC) in the MMT group (1.595±0.038) was significantly lower than that in the sham operation group (1.816±0.028) by 12.2% (p<0.05), while the bone mineral content in the C group (1.802±0.081), G group (2.071±0.057), and CLG group (2.068±0.058) was significantly higher than that in the sham operation group (1.595±0.038) by 13.0% (p<0.05), 29.9% (p<0.0001), and 29.7% (p<0.0001), respectively. Compared with the sham group (1.899±0.080), the anisotropy (DA) of the MMT group (1.292±0.058) was significantly reduced by 31.9% (p<0.005), while that of the CLG group (2.058±0.218) was increased by 59.3% (p=0.0003) compared with the MMT group.

[0090] The preparation method of the health food tablets of the present invention comprises the following steps: preparing glucosamine, turmeric powder, microcrystalline cellulose, sodium carboxymethyl starch, sodium cross-linked carboxymethyl cellulose and magnesium stearate; weighing glucosamine hydrochloride and turmeric powder after passing through a 60-mesh sieve for standby use; adding all the raw materials together into a blender for mixing, and blending by stirring; transferring the prepared granules into a dryer for drying; setting a tablet press, and compressing the tablets at a suitable speed to control the tablet weight; placing the compressed plain tablets in a coating pot, turning on a spray gun, and uniformly coating the plain tablets with a coating film to successfully prepare the tablets.

[0091] The preparation of the health food capsule of the present invention comprises the following steps: preparing glucosamine, turmeric powder, microcrystalline cellulose, magnesium stearate and silicon dioxide; weighing glucosamine hydrochloride and turmeric powder after passing through an 80-mesh sieve for standby use; adding all the raw materials together into a blender for mixing, stirring and granulating; transferring the prepared granules into a dryer for drying; filling the total mixed powder into capsules, and strictly controlling the product within an average capsule weight range during the filling process; polishing the filled capsules to make the capsule surfaces smooth and clean, and the capsules are successfully prepared.

[0092] The present invention proposes a composition for regulating inflammatory factors and improving subchondral bone damage, wherein glucosamine and turmeric powder are used as main effective ingredients, wherein the mass ratio of glucosamine to turmeric powder is 3:1. The prepared composition is verified by cell tests and animal tests to have the efficacy of regulating the level of inflammatory factors and improving subchondral bone damage. The composition of the present invention can be used as a main ingredient in foods that relieve joint discomfort by regulating the level of inflammatory factors and improving subchondral bone damage, and can be made into capsules, compressed candies, etc., and has broad application prospects.

[0093] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the present invention.

Claims

1. A composition for improving bone and joint health, characterized in that: Including glucosamine and turmeric powder.

2. The composition according to claim 1, characterized in that: In terms of mass ratio, the mass ratio of the turmeric powder to the glucosamine is 1:(0.2-5).

3. The composition according to claim 1 or 2, characterized in that: The mass ratio of the turmeric powder to glucosamine is 1:(0.3-3).

4. The composition according to claim 3, characterized in that: The content of curcumin in the turmeric powder is ≥1wt%.

5. A composition as claimed in any one of claims 1, 2 or 4, characterized in that: The composition regulates inflammatory factors and / or improves subchondral bone damage.

6. The composition according to claim 5, characterized in that: The inflammatory factors include one or more of NO, MMP13, TNF-α and IL-6.

7. Use of the composition according to any one of claims 1 to 6 in the preparation of foods, health foods and medicines having the effects of regulating inflammatory factors and improving subchondral bone damage.

8. The use according to claim 7, characterized in that: The inflammatory factors include one or more of NO, MMP13, TNF-α and IL-6.

9. A food, characterized in that: The food comprises the composition according to any one of claims 1 to 6 and a food-acceptable auxiliary material.

10. The food according to claim 9, characterized in that: The dosage form of the food includes tablets, capsules, drinks, soft candies or powders.

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