Composition for skeletal joint maintenance and preparation method and application thereof

Abstract

The invention relates to the technical field of functional food, and particularly discloses a composition for bone joint maintenance and a preparation method and application thereof. The composition for bone joint maintenance is prepared from 30 to 60 parts of a composite hydrogen-rich material and 30 to 60 parts of non-denatured type II collagen, the composite hydrogen-rich material comprises the following raw materials: silicon dioxide, calcium carbonate, magnesium oxide and hydrogen in a mass ratio of (60-80): (5-10): (1-10): (5-10). The composition provided by the invention improves blood circulation, accelerates absorption of necrotic tissues, eliminates edema, rapidly diminishes inflammation and relieves bone discomfort through the synergistic effect of all the components, and has good prevention and improvement effects on various skeletal system diseases. Functional food for promoting the health of the skeletal system is prepared from the composition, the application range is wide, good economic and social benefits are achieved, and the application prospect is wide.

Description

Technical Field

[0001] The present invention relates to the technical field of functional foods, and in particular to a composition for bone and joint care, and a preparation method and application thereof. Background Art

[0002] Among the many systems of the human body, the skeletal system plays a vital role. It is not only the basis for us to stand, walk and move, but also a key structure to protect internal organs, maintain body shape and participate in important physiological processes. With the development of modern society and changes in population structure, bone and joint health issues have gradually become the focus of global attention. Especially in the context of an aging society, the incidence of bone and joint diseases such as osteoarthritis and osteoporosis has increased year by year, seriously affecting the quality of life of middle-aged and elderly people. In addition, with the increasing awareness of sports and fitness among young people, joint problems caused by sports injuries have become increasingly prominent. According to data released by the World Health Organization, the number of people suffering from osteoarthritis worldwide has exceeded 500 million, and the incidence of arthritis is also becoming younger and younger. For example, gouty arthritis used to occur at an age of over 40 years old, but in recent years, due to changes in dietary structure, the age of onset has become younger and younger, and clinically, even teenage boys have developed gouty arthritis.

[0003] Currently, bone and joint care products on the market are mainly based on traditional ingredients such as glucosamine, chondroitin sulfate, calcium and vitamin D, and are mostly in the form of tablets, capsules and oral solutions. These products can relieve joint discomfort to a certain extent, but they have problems such as single ingredients, limited effects, slow onset and long treatment courses. Therefore, it is necessary to develop a new bone care composition to promote bone health and effectively prevent or treat diseases related to the skeletal system. Summary of the invention

[0004] In view of the problems of existing products for bone and joint maintenance, such as single ingredients, limited effects, slow onset of effect and long treatment course, the present invention provides a composition for bone and joint maintenance, a preparation method and application thereof.

[0005] In order to solve the above technical problems, the technical solution provided by the present invention is:

[0006] A composition for bone and joint care, comprising the following raw material components in parts by weight: 30 to 60 parts of a composite hydrogen-rich material and 30 to 60 parts of non-denatured type II collagen;

[0007] The raw materials of the composite hydrogen-rich material include silicon dioxide, calcium carbonate, magnesium oxide and hydrogen in a mass ratio of (60-80):(5-10):(1-10):(5-10).

[0008] Compared with the prior art, the composition for bone joint care provided by the present invention adds a composite hydrogen-rich material as an active component, and the composite hydrogen-rich material can improve the bioelectric level around the bone joint, provide an environment conducive to the growth and differentiation of bone cells, and promote the regeneration of bone tissue; at the same time, the composite hydrogen-rich material can promote angiogenesis, provide necessary nutrition and oxygen for new bone tissue, and accelerate the repair process, and the specific composite hydrogen-rich material of the present invention can inhibit inflammatory reactions, which has a positive effect on reducing the inflammatory process after bone injury and promoting bone healing; non-denatured type II collagen can relieve joint pain and inflammation, and can activate chondrocytes, promote the synthesis and repair of cartilage matrix, improve the structure and toughness of articular cartilage, and enhance joint mobility and flexibility; at the same time, the composite hydrogen-rich material can improve the repair efficiency of non-denatured type II collagen for cartilage tissue, effectively alleviate the discomfort symptoms of the skeletal system, and effectively inhibit the formation and development of osteoarthritis. The composition of the present invention can be used to prepare functional foods that promote the health of the skeletal system, with a wide range of applications, good economic and social benefits, and broad application prospects.

[0009] Furthermore, the composition for bone and joint care also includes: 18 to 36 parts of spiral flower, 6 to 12 parts of galangal powder and 18 to 36 parts of chrysanthemum notoginseng.

[0010] The spiral flower has the effects of lowering qi and resolving phlegm, lowering adverse qi and stopping vomiting, eliminating lumps and dispersing knots, clearing away heat and detoxifying, and relaxing muscles and activating blood circulation. The spiral flower is rich in nutrients, including flavonoids, chlorogenic acid and caffeic acid, which have antioxidant, anti-inflammatory and antibacterial effects, helping to reduce the inflammatory response caused by bone damage or disease, and creating a more favorable environment for the repair process. At the same time, the spiral flower also helps to improve blood circulation, thereby promoting bone repair and regeneration.

[0011] Galangal has the effects of warming the stomach and dispelling cold, helping digestion and sobering up. Galangal contains active ingredients such as volatile oils and galangin, which have analgesic effects.

[0012] Panax notoginseng is a medicinal and edible herb that has the effects of promoting bone development, anti-virus, anti-inflammatory and analgesic. At the same time, Panax notoginseng contains a large amount of natural organic calcium, which is easy for the human body to absorb and utilize, which helps promote bone development and prevent osteoporosis and bone and joint diseases.

[0013] The combined use of spiral flower, galangal and chrysanthemum notoginseng with composite hydrogen-rich materials and non-denatured type II collagen can effectively inhibit inflammatory response and promote the regeneration of cartilage tissue, thereby achieving the purpose of relieving pain, removing blood stasis and quickly repairing articular cartilage.

[0014] Furthermore, the composition for bone and joint care also includes: 12 to 24 parts of turmeric powder, 10 to 20 parts of acerola powder and 5 to 10 parts of seaweed calcium powder.

[0015] Curcumin is a natural polyphenol compound extracted from turmeric. It has powerful antioxidant properties, can neutralize free radicals, reduce oxidative stress, protect bone cells from damage, and has a positive effect on promoting bone health and repair. At the same time, curcumin can also regulate the immune system, which is beneficial to maintaining the immune balance during bone repair. It also has a good anti-inflammatory effect and helps relieve pain caused by bone problems. In addition, curcumin also helps to improve blood circulation, promote the proliferation and differentiation of bone cells, and increase bone density, thereby contributing to the repair and regeneration of bone tissue.

[0016] Acerola cherry powder can promote the synthesis of collagen, which is the main component of bones and cartilage, and helps with bone repair and regeneration. Acerola cherry powder can effectively prevent bone loss. The high levels of nutrients it contains (such as potassium, vitamin C, trace elements and fiber) can provide comprehensive nutritional support for bone repair.

[0017] Seaweed calcium powder can easily dissolve and ionize quickly in the human stomach and intestines, thereby increasing the body's absorption rate of calcium. At the same time, the various minerals in seaweed calcium powder can help increase bone density and prevent osteoporosis.

[0018] Galangal can improve the rapid blood stasis-removing effect of turmeric, and spiral flower can promote the analgesic effect of chrysanthemum and notoginseng. Turmeric, galangal, spiral flower and chrysanthemum and notoginseng synergize to effectively improve the anti-inflammatory and analgesic effects of the combination.

[0019] Furthermore, the mass ratio of the composite hydrogen-rich material to the non-denatured type II collagen is 1:1.

[0020] Furthermore, the mass ratio of the spiral flower with visible veins to the composite hydrogen-rich material and the total amount of non-denatured type II collagen is 10:3.

[0021] Furthermore, the mass ratio of the galangal powder to the total amount of the composite hydrogen-rich material and the non-denatured type II collagen is 10:1.

[0022] Furthermore, the mass ratio of the procumbent chrysanthemum notoginseng to the composite hydrogen-rich material and the total amount of non-denatured type II collagen is 10:3.

[0023] Furthermore, the mass ratio of the turmeric powder to the total amount of the composite hydrogen-rich material and the non-denatured type II collagen is 10:2.

[0024] Furthermore, the content of turmeric extract in the turmeric powder is not less than 98%.

[0025] Furthermore, the VC content in the acerola cherry powder is not less than 25%.

[0026] Furthermore, the composite hydrogen-rich material also includes potassium citrate.

[0027] Furthermore, the mass ratio of potassium citrate to silicon dioxide is (5-10):(60-80).

[0028] Furthermore, the preparation method of the composite hydrogen-rich material comprises the following steps:

[0029] S1, calcining the silicon dioxide at 700° C. to 800° C. for 1 h to 3 h under an inert atmosphere to obtain primary silicon dioxide;

[0030] S2, calcining the calcium carbonate at 800° C. to 900° C. for 1 h to 3 h under an inert atmosphere to obtain primary calcium carbonate;

[0031] S3, calcining the magnesium oxide at 500° C. to 600° C. for 1 h to 3 h under an inert atmosphere to obtain primary magnesium oxide;

[0032] S4, mixing the primary silicon dioxide, primary calcium carbonate and primary magnesium oxide uniformly, raising the temperature to 1300min-1400min, calcining for 30min-90min in a hydrogen atmosphere, cooling, crushing and mixing uniformly to obtain the composite hydrogen-rich material.

[0033] Furthermore, the preparation method of the composite hydrogen-rich material further comprises adding the potassium citrate to the composite hydrogen-rich material, mixing evenly, and crushing to 300-700 meshes to obtain the composite hydrogen-rich material.

[0034] Exemplarily, the particle size of the silicon dioxide is 100 μm to 300 μm, the median particle size of the calcium carbonate powder is 1 μm to 100 μm, and the median particle size of the magnesium oxide powder is 1 μm to 100 μm.

[0035] The present invention also provides a method for preparing a composition for bone and joint care, comprising the following steps:

[0036] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0037] The preparation method of the composition for bone and joint maintenance provided by the present invention has simple process, strong operability and is suitable for large-scale production and application.

[0038] The present invention also provides the use of the above composition for bone and joint care in the preparation of bone and joint care products.

[0039] The present invention also provides a functional food for bone and joint care, comprising any one of the above-mentioned compositions for bone and joint care.

[0040] The functional food for bone joint care provided by the present invention introduces a composite hydrogen-rich material as an active component, and the composite hydrogen-rich material can improve the bioelectric level around the bone joint, thereby improving the adsorption capacity of bone tissue to positively charged molecules such as calcium, magnesium, and phosphorus, and promoting the proliferation and differentiation of cells in the bone tissue; at the same time, a variety of active components such as non-denatured type II collagen, spiral flower of visible veins, galangal, and flat chrysanthemum Panax notoginseng are introduced to provide comprehensive nutrition for bone tissue metabolism and regeneration, ensure adequate nutrition of bone joints, and improve blood circulation through the synergistic effect of various components, accelerate the absorption of necrotic tissue, eliminate edema, quickly reduce inflammation and relieve pain, relieve bone discomfort, and have good prevention and improvement effects for a variety of skeletal system diseases. The composition of the present invention is prepared into a functional food that promotes the health of the skeletal system, which has a wide range of applications, good economic and social benefits, and broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 It is a comparison chart of chondrocyte data of blank control group, model control group, positive control group and glucosamine chondroitin group in Experimental Example 1 of the present invention; among which, *P<0.05; **P<0.01; ***P<0.001;

[0042] Figure 2 This is a comparison chart of the chondrocyte data of the blank control group, model control group, positive control group and sample group of comparative example 2 in Experimental Example 1 of the present invention; where *P<0.05; **P<0.01; ***P<0.001;

[0043] Figure 3 This is a comparison chart of the chondrocyte data of the blank control group, model control group, positive control group and sample group of Example 1 in Experimental Example 1 of the present invention; where *P<0.05; **P<0.01; ***P<0.001;

[0044] Figure 4 This is the chondrocyte phenotype diagram of the blank control group in Experimental Example 1;

[0045] Figure 5 This is the chondrocyte phenotype diagram of the model control group in Experimental Example 1;

[0046] Figure 6 This is the chondrocyte phenotype diagram of the positive control group in Experimental Example 1;

[0047] Figure 7 This is the chondrocyte phenotype diagram of the glucosamine chondroitin group in Experimental Example 1;

[0048] Figure 8This is the chondrocyte phenotype diagram of the sample group of comparative example 2 in experimental example 1;

[0049] Fig. 9 This is the chondrocyte phenotype diagram of the sample group of Example 1 in Experimental Example 1; DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0051] The composite hydrogen-rich material used in the following examples is made from the following raw materials in parts by weight:

[0052] 60 parts of silicon dioxide (particle size of 100-300 μm), 10 parts of calcium carbonate (median particle size of 1-50 μm), 1 part of magnesium oxide (median particle size of 1-50 μm) and 10 parts of hydrogen;

[0053] The preparation method thereof comprises at least the following steps:

[0054] Calcine silicon dioxide at 800°C in an inert atmosphere for 1 hour to obtain primary silicon dioxide;

[0055] Calcium carbonate is calcined at 900°C in an inert atmosphere for 3 hours to obtain primary calcium carbonate;

[0056] Calcine magnesium oxide at 500°C in an inert atmosphere for 1 hour to obtain primary magnesium oxide;

[0057] The primary silicon dioxide, primary calcium carbonate and primary magnesium oxide are mixed evenly, heated to 1400°C at a heating rate of 10°C / min, calcined for 90 minutes in a hydrogen atmosphere, cooled to 30°C, crushed to 300 mesh and mixed evenly to obtain the product.

[0058] In the following embodiments, composite hydrogen-rich materials prepared by using other raw material ratios defined in the present invention can also be used. As long as they are within the scope defined in the present invention, comparable technical effects can be achieved.

[0059] Non-denatured type II collagen was purchased from Shandong Baolijia Biotechnology Co., Ltd.; spiral flower, galangal powder, chrysanthemum notoginseng, turmeric powder, and acerola powder were from Xi'an Dihong Biotechnology Co., Ltd.; seaweed calcium powder was from Shanghai Jiechun Biotechnology Co., Ltd.

[0060] Example 1

[0061] This embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 45 parts of composite hydrogen-rich material and 45 parts of non-denatured type II collagen.

[0062] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0063] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0064] The composition for bone and joint care prepared in this example was added into water at a concentration of 1 g / 50 mL. The hydrogen concentration in the water was detected by an instrument and found to be 10 ppm, with a negative potential of -500 mV.

[0065] Example 2

[0066] This embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 60 parts of composite hydrogen-rich material and 30 parts of non-denatured type II collagen.

[0067] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0068] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0069] Example 3

[0070] This embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 30 parts of composite hydrogen-rich material and 60 parts of non-denatured type II collagen.

[0071] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0072] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0073] Example 4

[0074] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 30 parts of composite hydrogen-rich material, 30 parts of non-denatured type II collagen, 18 parts of spiral flower, 6 parts of galangal powder and 18 parts of chrysanthemum notoginseng.

[0075] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0076] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0077] Example 5

[0078] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 60 parts of composite hydrogen-rich material, 30 parts of non-denatured type II collagen, 22 parts of spiral flower, 12 parts of galangal powder and 36 parts of chrysanthemum notoginseng.

[0079] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0080] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0081] Example 6

[0082] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 30 parts of composite hydrogen-rich material, 60 parts of non-denatured type II collagen, 36 parts of spiral flower, 10 parts of galangal powder and 27 parts of chrysanthemum notoginseng.

[0083] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0084] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0085] Example 7

[0086] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 30 parts of composite hydrogen-rich material, 30 parts of non-denatured type II collagen, 12 parts of spiral flower, 6 parts of galangal powder and 18 parts of chrysanthemum notoginseng, 12 parts of turmeric powder, 15 parts of acerola powder and 8 parts of seaweed calcium powder.

[0087] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0088] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0089] Example 8

[0090] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 60 parts of composite hydrogen-rich material, 30 parts of non-denatured type II collagen, 18 parts of spiral flower, 12 parts of galangal powder and 36 parts of chrysanthemum notoginseng, 18 parts of turmeric powder, 20 parts of acerola powder and 10 parts of seaweed calcium powder.

[0091] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0092] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0093] Example 9

[0094] The present embodiment provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 30 parts of composite hydrogen-rich material, 60 parts of non-denatured type II collagen, 24 parts of spiral flower, 10 parts of galangal powder and 24 parts of chrysanthemum notoginseng, 24 parts of turmeric powder, 10 parts of acerola powder and 5 parts of seaweed calcium powder.

[0095] The preparation method of the above-mentioned composition for bone and joint care comprises the following steps:

[0096] The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

[0097] Comparative Example 1

[0098] This comparative example provides a composition for bone and joint care, comprising a composite hydrogen-rich material and non-denatured type II collagen in different mass ratios, which are 5:1, 4:1, 3:1, 1:3, 1:4, and 1:5, respectively; the total mass fraction of the composite hydrogen-rich material and non-denatured type II collagen is 90 parts.

[0099] Compositions for bone and joint care were prepared in exactly the same manner as in Example 1, which will not be described in detail here.

[0100] Comparative Example 2

[0101] This comparative example provides a composition for bone and joint care, comprising the following raw material components in parts by weight: 45 parts of a composite material and 45 parts of non-denatured type II collagen.

[0102] The composite material comprises the following components in parts by mass: 60 parts of silicon dioxide (particle size is 100-300 μm), 10 parts of calcium carbonate (median particle size is 1-50 μm), and 1 part of magnesium oxide (median particle size is 1-50 μm).

[0103] The preparation method of the composite material is as follows: weighing each component according to the designed ratio, mixing them evenly, and obtaining the composite material.

[0104] Test Example 1

[0105] 1. Obtaining embryos:

[0106] One hour after feeding in the afternoon of the day before the experiment, healthy adult wild-type zebrafish were selected, half male and half female, and placed in the breeding tank, one pair in each tank, and separated magnetically by a partition. At 9:00 am the next day, the partition was removed, and embryos were collected after free mating for 1 hour. Normally developed fertilized eggs were selected and placed in a culture dish, and E3 culture medium was added. No more than 100 embryos per dish were placed in a biochemical incubator at 28.5℃±0.5℃ for incubation.

[0107] 2. Preliminary experiment:

[0108] 2.1 Preparation of test solution:

[0109] Prepare a 10 mg / mL test substance stock solution with purified water, and dilute it with e3 culture medium in equal proportions to a gradient concentration solution (increase in steps of 100 μg / mL, with the highest concentration not exceeding 1000 μg / mL) for later use.

[0110] 2.2 Test substance treatment

[0111] Randomly select wild-type zebrafish larvae with normal development at 3 dpf (3 days after fertilization) in a 6-well plate, with 30 in each well and 1 replicate in each group (to ensure the accuracy and reliability of the data). Without harming the zebrafish, remove the culture medium in the 6-well plate, then add the test solution of different concentrations prepared above to 3 mL, add e3 culture medium to the blank control group, cover the culture plate, and place it in a biochemical incubator at 28.5℃±0.5℃ in the dark for 72 hours. Observe with a stereo microscope and count the deaths of zebrafish. The concentration group at which no death occurs in zebrafish at 7 dpf is the NOEC concentration.

[0112] The test results showed that there was no significant difference between the sample with a concentration of 500 μg / mL and the blank control group. Therefore, 500 μg / mL is the maximum safe concentration.

[0113] 2.3 Experimental Grouping

[0114] Model control group: prednisone was added to the e3 culture medium to obtain a 75 μg / mL prednisone solution, and the zebrafish larvae were placed in the prednisone solution at 28.5°C±0.5°C in a biochemical incubator for 72 h to establish a zebrafish cartilage injury model.

[0115] Positive control group: prednisone and etidronate disodium were added to the e3 culture medium to obtain a positive control solution of 75 μg / mL prednisone and 15 μg / mL etidronate disodium. The zebrafish larvae were placed in the above positive control solution in a biochemical incubator at 28.5°C ± 0.5°C for 72 hours.

[0116] Sample test group: Prednisone and the test substance were added to the e3 culture medium to obtain a sample solution of 75 μg / mL prednisone and 500 μg / mL test substance. The zebra fry were placed in the above sample solution in a biochemical incubator at 28.5°C±0.5°C for 72 hours. The test substances were selected as glucosamine chondroitin, the composition prepared in Comparative Example 2, and the composition prepared in Example 1.

[0117] Blank control group: The zebrafish larvae were placed in E3 culture medium in a biochemical incubator at 28.5℃±0.5℃ for 72h.

[0118] According to the above test requirements, normally developed 3 dpf zebrafish larvae were selected, washed with E3 culture medium and randomly distributed into 6-well plates, with 30 fish in each well and 1 well in each group. The E3 culture medium in the 6-well plates was removed without harming the larvae, and the solutions required by different groups were added, and the plates were incubated in a biochemical incubator at 28.5℃±0.5℃ for 72 hours.

[0119] 2.4 Judgment Basis

[0120] On the premise of meeting the validity, the fluorescence intensity of the model control group is significantly reduced compared with the blank control group (P < 0.05), which proves that the modeling is successful. The fluorescence intensity of the sample test group is significantly enhanced compared with the model control group (P < 0.05), indicating that the sample has the effect of cartilage repair. The fluorescence intensity is photographed using a fluorescence microscope to obtain fluorescence intensity photos.

[0121] 2.5 Data processing

[0122] SPSS26 software was used for analysis, GraphPad Prism8 was used for drawing, and data statistics were expressed as mean ± standard deviation. SPSS software was used to test the normal distribution of the experimental results. If the data conformed to the normal distribution, the T parameter test or analysis of variance (ANOVA) was used for significance. If it did not conform to the normal distribution, a non-parametric test was used (the result P value < 0.05 indicated a statistically significant difference).

[0123] 2.6 Test Results

[0124] The results of the glucosamine chondroitin sample test group are shown in Table 1 and Figure 1 shown.

[0125] Table 1 Glucosamine Chondroitin Sample Test Group Results

[0126]

[0127] The results of the test group of the composition samples prepared in Comparative Example 2 are shown in Tables 2 and Figure 2 shown.

[0128] Table 2 Comparative Example 2 Sample Test Group Results

[0129]

[0130] The results of the test group of the composition samples prepared in Example 1 are shown in Tables 3 and Figure 3 shown.

[0131] Table 3 Example 1 Sample Test Group Results

[0132]

[0133]

[0134] 3. Chondrocyte phenotype map

[0135] Fluorescence microscopy was used to irradiate and sample the zebrafish of the above different treatment groups to obtain the whole body bone tissue cell level. The results are as follows Figures 4 to 9 shown.

[0136] As can be seen from the figure, the proportion of bone tissue cells in the sample treatment group of Example 1 was significantly increased compared with other groups, proving that the composition prepared in the example of the present invention can promote the proliferation and differentiation of chondrocytes.

[0137] Test Example 2

[0138] According to the method of Experimental Example 1, the cartilage repair efficacy test was performed using the composition prepared by using different ratios of the composite hydrogen-rich material and the non-denatured type II collagen in Comparative Example 1. After the incubation, the fluorescence intensity was obtained by taking pictures using a fluorescence microscope. The results are shown in Table 4.

[0139] Table 4 Fluorescence intensity of compositions with different ratios

[0140] Test Group Fluorescence intensity 5:1 38.21±2.17 4:1 39.21±3.27 4:1 40.11±2.36 2:1 (Example 2) 42.15±4.27 1:1 (Example 1) 48.00±10.71 1:2 (Example 3) 41.00±3.27 1:3 40.31±2.16 1:4 39.06±2.36 1:5 38.11±3.36

[0141] The results show that the cartilage tissue repair effects of the compositions prepared in Examples 1 to 3 are better than those of the compositions prepared in other proportions.

[0142] Test Example 3

[0143] This comparative example provides a composition for bone and joint care, including a composite hydrogen-rich material and non-denatured type II collagen (abbreviated as HB) in a mass ratio of 1:1, which are respectively combined with spiral flower of venous spiral, galangal, chrysanthemum notoginseng and turmeric powder in different mass ratios to test the analgesic effects of different compositions.

[0144] Among them, the mass ratios of HB and different other components are 10:1, 10:2, 10:3, 10:4, 10:5, 10:6, 10:7, 10:8, 10:9, 10:10, 9:10, 8:10, 7:10, 6:10, 5:10, 4:10, 3:10, 2:10, and 1:10, respectively.

[0145] The composition for bone and joint care was prepared in exactly the same manner as in Example 1, which will not be described again here.

[0146] Similarly, the composition for bone and joint care (abbreviated as HBB) prepared in the test comparison example 2 according to the above ratio was combined with spiral flower of visible vein, galangal, chrysanthemum notoginseng and turmeric powder in different mass ratios to test the analgesic effects of different compositions.

[0147] At 24 hpf, the zebrafish's nerves are not fully developed and it is unable to perform autonomous twitching movements. It will only twitch when it encounters external substances. Therefore, the number of twitches within a certain period of time can evaluate the intensity of external pain, and the analgesic effect of the sample can be judged by whether the number of twitches is reduced.

[0148] 3.1 Experimental Grouping:

[0149] Model control group: Pentylenetetrazol was dissolved in e3 culture medium to obtain a 50 μg / mL pentylenetetrazol solution. The zebrafish fry were placed in the pentylenetetrazol solution at 28.5°C±0.5°C in a biochemical incubator for 72 hours to establish a zebrafish convulsion model.

[0150] Sample test group: Add pentylenetetrazol and the test substance to the e3 culture medium to obtain a sample solution of 50 μg / mL pentylenetetrazol and 500 μg / mL test substance. Place zebra fry in the above sample solution at 28.5℃±0.5℃ biochemical incubator for 72 hours.

[0151] Blank control group: The zebrafish larvae were placed in E3 culture medium in a biochemical incubator at 28.5℃±0.5℃ for 72h.

[0152] According to the above test requirements, normally developed 24 hpf zebrafish larvae were selected, washed with E3 culture medium and randomly distributed into 6-well plates, with 30 fish in each well and 1 well in each group. The E3 culture medium in the 6-well plates was removed without harming the larvae, and the solutions required by different groups were added, and the plates were incubated in a biochemical incubator at 28.5°C±0.5°C for 72 hours.

[0153] 3.2 Data processing

[0154] SPSS26 software was used for analysis, GraphPad Prism8 was used for drawing, and data statistics were expressed as mean ± standard deviation. SPSS software was used to test the normal distribution of the experimental results. If the data conformed to the normal distribution, the T parameter test or analysis of variance (ANOVA) was used for significance. If it did not conform to the normal distribution, a non-parametric test was used (the result P value <0.05 indicated a statistically significant difference).

[0155] 3.3 Judgment basis

[0156] Compared with the blank control group, the number of involuntary twitches of zebrafish in the model test group was significantly higher than that in the blank control group (P<0.05), indicating that the modeling was successful.

[0157] Compared with the model control group, the number of involuntary twitches of zebrafish in the sample test group was significantly lower than that in the model control group (P<0.05), indicating that the sample has analgesic effect.

[0158] 3.4 Test Results

[0159] Table 5 HB+ different proportions of turmeric powder

[0160]

[0161]

[0162] Note: The above significant differences are compared with the model control group, P < 0.05.

[0163] Table 6 HBB + different proportions of turmeric powder

[0164]

[0165]

[0166] Note: The above significant differences are compared with the model control group, P < 0.05.

[0167] Table 7 HB+ different proportions of visible vein spiral flowers

[0168]

[0169]

[0170] Note: The above significant differences are compared with the model control group, P < 0.05.

[0171] Table 8 HBB+ different proportions of visible vein spiral flowers

[0172]

[0173]

[0174] Note: The above significant differences are compared with the model control group, P < 0.05.

[0175] Table 9 HB+ different proportions of galangal

[0176] Group Number of draws P-value Blank control group 3.43±0.90 Model control group 9.00±0.76 10:1 7.13±0.22 P<0.05 10:2 7.23±0.35 P<0.05 10:3 7.35±0.22 P<0.05 10:4 7.46±0.23 P<0.05 10:5 7.65±0.38 P<0.05 10:6 7.85±0.63 P>0.05 10:7 7.92±0.31 P>0.05 10:8 7.99±0.18 P>0.05 10:9 8.11±0.25 P>0.05 10:10 8.13±0.56 P>0.05 9:10 8.24±0.32 P>0.05 8:10 8.39±0.26 P>0.05 7:10 8.14±0.37 P>0.05 6:10 8.04±0.39 P>0.05 5:10 8.34±0.37 P>0.05 4:10 8.55±0.65 P>0.05 3:10 8.85±0.39 P>0.05 2:10 8.91±0.22 P>0.05 1:10 8.95±0.37 P>0.05

[0177] Note: The above significant differences are compared with the model control group, P < 0.05.

[0178] Table 10 HBB + different proportions of galangal

[0179]

[0180]

[0181] Note: The above significant differences are compared with the model control group, P < 0.05.

[0182] Table 11HB+ flat chrysanthemum Panax notoginseng

[0183]

[0184]

[0185] Note: The above significant differences are compared with the model control group, P < 0.05.

[0186] Table 12 HBB + flat chrysanthemum Panax notoginseng

[0187]

[0188]

[0189] Application Example 1

[0190] The composition prepared in Example 7 was tested for its pain relief and cartilage tissue repair effects according to the experimental procedures of Test Example 1 and Test Example 3, respectively. The results are shown in Table 13.

[0191] Table 13 Pain relief

[0192]

[0193] Table 14 Cartilage tissue repair

[0194]

[0195] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition for bone and joint care, It is characterized in that The raw material components include the following parts by weight: 30 to 60 parts of composite hydrogen-rich material and 30 to 60 parts of non-denatured type II collagen; The raw materials of the composite hydrogen-rich material include silicon dioxide, calcium carbonate, magnesium oxide and hydrogen in a mass ratio of (60-80):(5-10):(1-10):(5-10).

2. The composition for bone and joint care according to claim 1, It is characterized in that The composition for bone and joint care also includes: 18 to 36 parts of spiral flower, 6 to 12 parts of galangal powder and 18 to 36 parts of chrysanthemum notoginseng.

3. The composition for bone and joint care according to claim 2, It is characterized in that The composition for bone and joint care also includes: 12 to 24 parts of turmeric powder, 10 to 20 parts of acerola powder and 5 to 10 parts of seaweed calcium powder.

4. The composition for bone and joint care according to claim 3, It is characterized in that The mass ratio of the composite hydrogen-rich material to the non-denatured type II collagen is 1:1; and / or The mass ratio of the total amount of the helical flower of the veins, the composite hydrogen-rich material and the non-denatured type II collagen is 10:3; and / or The mass ratio of the galangal powder to the total amount of the composite hydrogen-rich material and the non-denatured type II collagen is 10:1; and / or The mass ratio of the flat prostrate chrysanthemum notoginseng to the composite hydrogen-rich material and the total amount of non-denatured type II collagen is 10:3; and / or The mass ratio of the turmeric powder to the composite hydrogen-rich material and the total amount of the non-denatured type II collagen is 10:

2.

5. The composition for bone and joint care according to claim 4, It is characterized in that The mass ratio of the potassium citrate to silicon dioxide is (5-10):(60-80).

6. The composition for bone and joint care according to claim 1, It is characterized in that The content of turmeric extract in the turmeric powder is not less than 98%; and / or The VC content in the acerola cherry powder is not less than 25%.

7. The composition for bone and joint care according to claim 1, It is characterized in that The preparation method of the composite hydrogen-rich material comprises the following steps: S1, calcining the silicon dioxide at 700° C. to 800° C. for 1 h to 3 h under an inert atmosphere to obtain primary silicon dioxide; S2, calcining the calcium carbonate at 800° C. to 900° C. for 1 h to 3 h under an inert atmosphere to obtain primary calcium carbonate; S3, calcining the magnesium oxide at 500° C. to 600° C. for 1 h to 3 h under an inert atmosphere to obtain primary magnesium oxide; S4, mixing the primary silicon dioxide, primary calcium carbonate and primary magnesium oxide uniformly, raising the temperature to 1300min-1400min, calcining for 30min-90min in a hydrogen atmosphere, cooling, crushing and mixing uniformly to obtain the composite hydrogen-rich material.

8. A method for preparing the composition for bone and joint care according to any one of claims 1 to 7, It is characterized in that The following steps are involved: The components are weighed according to the designed ratio, and the weighed components are dried and then mixed evenly to obtain a composition for bone and joint care.

9. Use of the composition for bone and joint care according to any one of claims 1 to 7 in the preparation of bone and joint care products.

10. A functional food for bone and joint care, It is characterized in that A composition for bone and joint care comprising the composition described in any one of claims 1 to 7.

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