Hemostatic composition and application thereof in oral care product

Abstract

The invention discloses a hemostatic composition which comprises fibronectin, a rhizoma bletillae extract and a radix notoginseng extract. The fibronectin, the rhizoma bletillae extract and the pseudo-ginseng extract are combined for use and have a synergistic effect, the rhizoma bletillae extract and the pseudo-ginseng extract promote fibrinogen to become insoluble fibrin by activating platelets and activating thrombin, and the fibronectin is effectively cross-linked with fibrin at the'inner core 'of the hemostatic plaque, so that platelet aggregation is promoted, and the hemostatic plaque has the function of promoting hemostasis. Therefore, the effect of quickly stopping bleeding is achieved. The hemostatic composition disclosed by the invention can be applied to oral care products, has remarkable gingival protecting and hemostatic effects, can effectively improve the problems of gingival bleeding and oral inflammation, and achieves the effects of easing pain and diminishing swelling.

Description

Technical Field

[0001] The invention relates to the technical field of oral care, and in particular to a hemostatic composition and application thereof in oral care products. Background Art

[0002] Bleeding gums are one of the common symptoms of oral diseases. They refer to a small amount of bleeding gums that occurs spontaneously or due to minor irritation. In mild cases, there will be blood in the saliva only when sucking, brushing teeth, or chewing hard food. In severe cases, the gums will bleed more or even spontaneously when they are slightly stimulated. Generally speaking, gingivitis or periodontitis is the most common cause of bleeding gums. When the gums are acutely or chronically inflamed, the gums will become congested and edematous, and the capillaries in the gums will dilate and proliferate. At this time, when brushing teeth, touching or chewing food, the capillaries are prone to rupture and bleeding gums will occur. According to the data from the "Fourth National Oral Health Epidemiology" survey report, the detection rate of gum bleeding is: 87.4% in the 35-44 age group, with an average of 13.77 teeth with bleeding gums per person; 88.45% in the 55-64 age group, with an average of 13.87 teeth with bleeding gums per person; 72.6% in the 65-74 age group, with an average of 11.25 teeth with bleeding gums per person. Gum bleeding is often accompanied by bad breath, which will affect work and social activities to a certain extent, and may even bring mental burden to patients. Studies have shown that if gum bleeding is not treated in time and bleeding frequently, it may be prone to many complications, such as chronic anemia. Therefore, sufficient attention should be paid to the problem of gum bleeding.

[0003] At present, the most important measure for gum bleeding is prevention. Developing good oral hygiene habits can effectively reduce the risk of gum bleeding. Using oral care products that protect gums and stop bleeding (such as toothpaste) is an important auxiliary means in daily oral care.

[0004] In recent years, the toothpaste industry has developed towards specialization, providing professional solutions for different dental problems, reflecting the continued upgrading of consumers' attention to oral health. At present, there are many gum-protecting and hemostatic toothpaste products on the market, but their gum-protecting and hemostatic effects vary. Some of them have drug resistance problems and are not suitable for long-term use. There are not many options for consumers, and gum protection technology is difficult to upgrade. Therefore, it is urgent to develop an oral care product that can effectively improve gum bleeding and oral inflammation problems to meet consumers' daily oral care needs. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a hemostatic composition, which has the effect of rapid hemostasis, can be applied to oral care products, can effectively improve gum bleeding and oral inflammation problems, and achieve the effect of analgesia and swelling reduction.

[0006] The present invention is achieved through the following technical solutions:

[0007] A hemostatic composition comprises fibronectin, a Bletilla striata extract and a Panax notoginseng extract.

[0008] The fibronectin described in the present invention, namely fibronectin (Fn), is a high molecular weight glycoprotein in the extracellular matrix.

[0009] The present invention uses fibronectin in combination with Bletilla striata extract and Panax notoginseng extract, and the three have a synergistic effect. The Bletilla striata extract and Panax notoginseng extract promote the conversion of fibrinogen into insoluble fibrin by activating platelets and activating thrombin. The fibronectin effectively cross-links with the fibrin in the "inner core" of the hemostatic plaque, promotes platelet aggregation, and thus achieves the effect of rapid hemostasis.

[0010] Preferably, the weight ratio of fibronectin, Bletilla striata extract and Panax notoginseng extract is 1:(1-5000):(1-1000), more preferably 1:(2-400):(1-120), and even more preferably 1:(4-200):(1-40). When the ratio of fibronectin or Panax notoginseng extract is too high, while the fibronectin in the "core" of the thrombus is covalently linked to fibrin, the non-fibrin on the periphery of the thrombus will be linked to the fibronectin, inhibiting the further growth of the thrombus, preventing vascular occlusion, and maintaining the blood supply to downstream tissues and organs, which in turn affects the hemostatic effect.

[0011] Preferably, the weight content of Bletilla striata polysaccharide in the Bletilla striata extract is 1.94-2.2%, and the weight average molecular weight of the Bletilla striata polysaccharide is 1000DA-30KDA.

[0012] The Bletilla striata extract of the present invention can be obtained by commercial purchase or homemade. As a preferred embodiment, the present invention provides a preparation method of the Bletilla striata extract, comprising the following steps: soaking the Bletilla striata tuber raw material in 45-55°C water for 4-5h, the solid-liquid ratio is 1: (6-40), after crushing, ultrasonic extraction is performed at a temperature of 55-65°C for 50-80min, wherein the ultrasonic power is 120-450w, and then water and ethanol are added (preferably, based on the mass of the Bletilla striata tuber raw material as 1 part by mass, 6-40 parts by mass of ethanol are added, and the ratio of ethanol to water is 1: 1), reflux extraction at 55-65°C for 120-150min, filtering, collecting the filtrate, and adding 80-90°C hot water to the filter residue for extraction again, the liquid-to-solid ratio is 1: (45-50), ultrasonic extraction for 50-70min, filtering, collecting the filtrate, combining the filtrate, concentrating the obtained filtrate at a temperature of 85-95°C, filtering and collecting the filtrate for high-pressure sterilization, and obtaining the Bletilla striata extract. The filtration may be performed using a 1 μm filter membrane; after the filtrate is sterilized, a preservative and water may be added to prepare a Bletilla striata extract. Preferably, the Bletilla striata tuber raw material is a dried Bletilla striata tuber. More preferably, the Bletilla striata tuber raw material is free of wormholes, mildew, and uniform in size.

[0013] As a preferred embodiment, the drying method of Bletilla striata tubers includes the following steps: after digging up the Bletilla striata tubers, the fibrous roots are removed, washed, boiled or steamed in boiling water until there is no white core, then dried in the sun until half dry, the outer skin is removed, and further dried in the sun to obtain a dry Bletilla striata tuber raw material.

[0014] Preferably, the weight content of total notoginseng saponins in the notoginseng extract of the present invention is not less than 75%.

[0015] The Panax notoginseng extract of the present invention can be obtained by commercial purchase or homemade. As a preferred embodiment, the present invention provides a preparation method of the Panax notoginseng extract, comprising the following steps: after the Panax notoginseng raw material is crushed, 70%-75% ethanol is added, the solid-liquid ratio is 1: (12-15), reflux extraction is performed at a temperature of 60-70°C for 40-60 minutes, concentrated by freeze drying to obtain a concentrated extract, dissolved in water, filtered, 80% ethanol is added to the filtrate, and then concentrated by adsorption resin to obtain a second concentrated extract, then water and 90-95% ethanol are added to dissolve, then activated carbon is added for decolorization, and then a filtrate is obtained by filtering with a titanium rod, refined and concentrated to obtain an extract, the extract is vacuum dried, crushed into fine powder, sieved, and the Panax notoginseng extract is obtained.

[0016] The adsorption resin is preferably D101 macroporous adsorption resin, which is washed with water until the effluent is colorless, then eluted with 50% ethanol, and the elution endpoint is checked by TLC to obtain a second concentrated extract.

[0017] Preferably, the raw material of Panax notoginseng is dried rhizomes of Panax notoginseng, and more preferably, the raw material of Panax notoginseng rhizomes is free of wormholes, mildew and uniform in size.

[0018] As a preferred embodiment, the drying method of Panax notoginseng rhizomes comprises the following steps:

[0019] Take fresh Panax notoginseng roots, wash them, remove the rhizomes and fibrous roots, keep the main root, then cut the main root into two halves, and then freeze-dry (-60-50°C, -0.32-0.36MPa) for 1-4 days until the moisture mass fraction is less than 10%.

[0020] The present invention also provides application of the hemostatic composition in oral care products.

[0021] Preferably, the mass content of the hemostatic composition in the oral care product is 0.6%.

[0022] Preferably, the oral care product comprises toothpaste, mouthwash or mouth spray.

[0023] The present invention has the following beneficial effects:

[0024] The hemostatic composition of the present invention uses fibronectin in combination with Bletilla striata extract and Panax notoginseng extract, and the three have a synergistic effect and have a rapid hemostatic effect.

[0025] The hemostatic composition of the present invention can be applied to oral care products, has significant gum protection and hemostasis effects, can effectively improve gum bleeding and oral inflammation problems, achieve the effects of analgesia and swelling reduction, and meet people's daily oral care needs. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The fibronectin content in the mice of the radiotherapy-induced injury group and the normal-induced injury group;

[0027] Figure 2 The skin healing speed of the radiotherapy-induced injury group and the normal-induced injury group;

[0028] Figure 3 The skin healing speed of the wounds applied with the hemostatic composition and the control gel respectively; DETAILED DESCRIPTION

[0029] In order to elaborate on the technical content, purpose and effect of the present invention, the technical solution of the present invention is clearly and completely described in combination with the implementation mode and the accompanying drawings below, but the described embodiments are only part of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] The raw materials used in the embodiments of the present invention are described as follows, but are not limited to the following raw materials:

[0031] Fibronectin: Fibronectin from Qidian Biotechnology, 500 ppm fibronectin;

[0032] Bletilla striata extract: contains about 2% Bletilla striata polysaccharide (Bletilla striata gum), with a molecular weight of 1000DA-30KDA;

[0033] The method is as follows: after the tubers of Bletilla striata are dug, the fibrous roots are removed, washed, steamed in boiling water until there is no white core, then sun-dried until half-dried, the outer skin is removed, and then sun-dried to obtain a dried tuber material of Bletilla striata. Then, qualified tuber materials of Bletilla striata (no worm eyes, no mildew, and uniform size) are soaked in 50°C water for 4 hours, with a material-liquid ratio of 1:40, and after crushing, ultrasonic extraction is performed at a temperature of 60°C for 80 minutes, wherein the ultrasonic power is 450w, and then appropriate amounts of water and ethanol are added, reflux extraction is performed at 60°C for 120 minutes, filtered, the filtrate is collected, and the filter residue is added with 85°C hot water for extraction again, with a liquid-to-material ratio of 1:45, ultrasonic extraction is performed for 50 minutes, filtered, the filtrate is combined, the obtained filtrate is concentrated at a temperature of 85°C, filtered with a 1μm filter membrane, and sterilized by high pressure, and the filtrate is collected, and a preservative and water are added to obtain a Bletilla striata extract.

[0034] Panax notoginseng extract: the total saponin content of Panax notoginseng is more than 75%;

[0035] It is prepared by the following method: high-quality Panax notoginseng raw materials (consistent size, no worm eyes, no mildew) are screened from the dried roots and rhizomes of Panax notoginseng, crushed, added with 70% ethanol, the solid-liquid ratio is 1:12, reflux extraction is carried out at a temperature of 60°C for 50 minutes, concentrated by freeze-drying to obtain a concentrated extract, dissolved in water, filtered, 80% ethanol is added to the filtrate, D101 macroporous adsorption resin is used, washed with water until the effluent is colorless, eluted with 50% ethanol, and the elution end point is checked by TLC to obtain a second concentrated extract, then water and 95% ethanol are added to dissolve, and then activated carbon is added for decolorization, and then a filtrate is obtained by filtering with a titanium rod, which is refined and concentrated to obtain an extract, the extract is vacuum dried, crushed into fine powder, sieved, and a Panax notoginseng extract is obtained.

[0036] 1. Evaluation of hemostatic effect

[0037] The bioactivity assay of thrombin time was adopted. The specific steps are as follows: 0.1 mL of the pig plasma to be tested (pig plasma is obtained by aseptic blood collection, EDTA anticoagulant is added, and it can be stored at -18°C for one year, Guangzhou Hongquan Biotechnology Co., Ltd.) was taken in a 2 ml centrifuge tube, and then 0.1 ml of the test agent (deionized water was used to dilute the test sample) was added, and 0.1 mL of normal saline was added to the blank control, mixed, incubated in a 37°C water bath for 3 min, 0.1 mL of pig thrombin was added, and mixed by vortex oscillation for 1 s, and the stopwatch was immediately started until the color of the sample liquid turned white and turbid and the liquid surface of the centrifuge tube did not flow when it was inverted. Record the coagulation time, which is the TT value (s) (TT value refers to the time for blood to coagulate after the standardized thrombin is added to the plasma). Each sample was repeated 10 times and the average value was taken. Statistical analysis, the experimental data are expressed as mean ± standard deviation (±S), and the experimental results are shown in Table 1 below.

[0038] Table 1 Content of each component (wt%) of the hemostatic composition to be tested in Examples 1-5 and its hemostatic effect data

[0039] Example 1 Example 2 Example 3 Example 4 Example 5 Fibronectin 0.2000 0.1000 0.0100 0.0025 0.0001 Bletilla Extract 0.2000 0.4000 0.4900 0.4975 0.4999 Panax Notoginseng Extract 0.2000 0.1000 0.1000 0.1000 0.1000 TT value / s 10.5±0.96 6.8±1.03 8.0±0.94 7.4±0.70 9.0±0.94

[0040] Table 2 Comparative Examples 1-8 Hemostatic Composition Test Agents Component Content (wt%) and Hemostatic Effect Data

[0041]

[0042]

[0043] Table 2

[0044] Comparative Example 6 Comparative Example 7 Comparative Example 8 Fibronectin / 0.6000 / Bletilla Extract 0.6000 / / Panax Notoginseng Extract / / 0.6000 TT value / s 11.20±0.85 15.10±0.99 15.90±0.57

[0045] It can be seen from the above test results that the hemostatic composition of the present invention, by combining fibronectin with Bletilla striata extract and Panax notoginseng extract, has a synergistic effect and has the effect of rapid hemostasis.

[0046] In Comparative Example 1-2, the proportion of fibronectin or Panax notoginseng extract is too high, and the hemostatic effect is poor.

[0047] It can be seen from Comparative Examples 3-8 that the composition without fibronectin, Bletilla striata extract or Panax notoginseng extract has poor hemostatic effect, which shows that the present scheme can produce obvious synergistic effect in hemostatic effect by combining the three substances.

[0048] 2. Mouse Wound Healing Model Experiment

[0049] Sixteen healthy and normal BALb / c mice aged 4 to 6 weeks were divided into 4 groups. Group 1 was the radiotherapy-injured group without treatment, Group 2 was the normal injury control group, Group 3 was the radiotherapy-injured control group, and Group 4 was the radiotherapy-injured group treated with the hemostatic composition. The other three groups except Group 2 were irradiated with 20Gy radiation on the left ventral side and rested for four weeks. The other three groups except Group 2 were subjected to 8mm puncture biopsy injury at the irradiated site. Group 2 was subjected to normal 8mm puncture biopsy injury and stent implantation. Then Group 1 and Group 2 were not treated, Group 3 was treated with the control gel, and Group 4 was treated with the hemostatic composition of Example 1. The wound area was measured regularly at the wound until the wound closed. See the test results. Figure 1 , Figure 2 and Figure 3 .

[0050] from Figure 1 and Figure 2 It can be seen that the skin wounds of mice receiving radiotherapy recovered more slowly than those of the normal group, so radiotherapy was selected as the method of injuring mice. Figure 3 It can be seen that the hemostatic composition of the present invention can accelerate the healing speed of radiotherapy skin wounds.

[0051] 3. Oral Care Product Application Test

[0052] Preparation of toothpaste: According to the proportions in Table 3,

[0053] S1: dissolving the hemostatic composition, dipotassium glycyrrhizinate, sodium saccharin, sodium fluoride, sodium benzoate, PEG400, and sorbitol in water to form a solution, and then adding titanium dioxide to disperse evenly;

[0054] S2: Mix hydrated silica, thickener and K12 evenly, add the solution in S1, stir evenly in a vacuum to form a paste;

[0055] S3: dissolving the cooling agent with essence, and adding it and sodium lauroyl sarcosinate into the paste of S2, stirring evenly under vacuum, to prepare a gum protecting and hemostatic toothpaste containing the hemostatic composition.

[0056] Table 3 Component formula of gum protection and hemostatic toothpaste (wt%)

[0057]

[0058] Toothpaste product hemostasis test verification

[0059] The toothpastes prepared in Application Examples 1-3 and Comparative Example 1 were taken as experimental samples.

[0060] New Zealand rabbits that have passed quarantine were selected, weighing 2.0kg to 2.5kg, female, and divided into 4 groups, with 10 rabbits in each group. 30mg / kg of 3% sodium pentobarbital was slowly injected through the ear vein. After the rabbits were anesthetized successfully, the abdomen was opened layer by layer to expose the liver, and a 1cm*1cm*3cm wound was clamped at the lower edge of the left lobe of the liver in the same part of each animal. After the wound is filled with blood, 1.0g of the test sample was immediately applied to the entire wound, covered with gauze, and a 50g weight was used to apply pressure on the gauze to observe the hemostatic effect. The bleeding of the wound was observed every 30s, and the filter paper strip was gently dipped and absorbed until the blood stopped oozing out, that is, there was no blood on the filter paper strip, and the required time was recorded as the bleeding time. If the bleeding has not stopped for more than 12min, press to stop the bleeding, and the bleeding time is counted as 12min. The animals were euthanized after the experiment. For statistical analysis, the experimental data are expressed as mean ± standard deviation The experimental results are shown in Table 4 below.

[0061] Table 4 Hemostatic effect data of application examples 1-3 and application comparative example 1

[0062] Hemostasis time (min) Application Example 1 6.25±0.42 Application Example 2 5.90±0.40 Application Example 3 6.17±0.30 Application Comparative Example 1 7.54±0.24

[0063] Verification of anti-inflammatory effects of toothpaste products

[0064] The toothpastes prepared in Application Examples 1-3 and Application Comparative Example 1 were used to conduct an experiment on the effect of p-xylene on the ear swelling of mice to study their anti-inflammatory effects. The deionized water and dexamethasone sodium phosphate injection group were used as the control group.

[0065] Take SPF KM mice that have passed the quarantine, 4 groups, 10 mice in each group, SPF KM mice, 18-22g, male. After the mice were quarantined and grouped, the left ear was not treated as the background value, and 30μL of xylene solution was applied to the inside and outside of the right ear. After 30min, except for the model control group, 0.1g of sample was applied to both sides of the right ear of the mice in the test sample group, and 50μL of dexamethasone sodium phosphate injection was applied to the dexamethasone sodium phosphate injection group. The injection was applied for the second time 30min later. After 1h of sample application, the mouse eyeball was removed to collect blood, and the mouse was killed by cervical dislocation, the sample was wiped clean, the ears were cut off, and the ear pieces with a diameter of 8mm (the same part of the left and right ears) were weighed and recorded. The weight difference between the left and right ears of the same animal is the swelling degree.

[0066] Swelling degree = weight of right ear piece - weight of left ear piece

[0067] Swelling inhibition rate = (average swelling degree of model control group - average swelling degree of treatment group) / average swelling degree of model control group * 100%)

[0068] The test results are shown in Table 5 below.

[0069] Table 5 Anti-inflammatory effect data of application examples 1-3 and application comparative example 1

[0070] Group Ear swelling (mg) Ear swelling inhibition rate (%) Model control group 21.3±3.7 - Application Example 1 9.8±2.5 53.99 Application Example 2 7.2±3.3 66.20 Application Example 3 12.1±5.0 43.19 Application Comparative Example 1 20.7±2.9 2.82 Dexamethasone sodium phosphate injection group 15.4±4.3 27.70

[0071] It can be seen from the above results that the hemostatic composition of the present invention can be applied to oral care products, has significant gum protection and hemostasis effects, can effectively improve gum bleeding and oral inflammation problems, and achieve the effect of analgesia and swelling reduction.

[0072] The above descriptions are merely embodiments of the present invention and are not intended to limit the patent scope of the present invention. Any equivalent transformations made using the contents of the present invention's specification and drawings, or directly or indirectly applied in related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A hemostatic composition, characterized in that: Including fibronectin, Bletilla striata extract and Panax notoginseng extract.

2. The hemostatic composition according to claim 1, characterized in that The weight ratio of fibronectin, Bletilla striata extract and Panax notoginseng extract is 1:(1-5000):(1-1000).

3. The hemostatic composition according to claim 1, characterized in that The weight ratio of fibronectin, Bletilla striata extract and Panax notoginseng extract is 1:(2-400):(1-120).

4. The hemostatic composition according to claim 1, characterized in that The weight ratio of fibronectin, Bletilla striata extract and Panax notoginseng extract is 1:(4-200):(1-40).

5. The hemostatic composition according to claim 1, characterized in that The weight content of the bletilla striata polysaccharide in the bletilla striata extract is 1.94-2.2%, and the weight average molecular weight of the bletilla striata polysaccharide is 1000DA-30KDA; the weight content of the notoginseng total saponin in the notoginseng extract is not less than 75%.

6. The hemostatic composition according to claim 1, characterized in that The preparation method of the bletilla striata extract comprises the following steps: soaking the bletilla striata tuber raw material in 45-55°C water for 4-5h, with a solid-liquid ratio of 1:(6-40), crushing, performing ultrasonic extraction at a temperature of 55-65°C for 50-80min, wherein the ultrasonic power is 120-450w, then adding water and ethanol, reflux extraction at 55-65°C for 120-150min, filtering, collecting the filtrate, and adding 80-90°C hot water to the filter residue for extraction again, with a liquid-to-solid ratio of 1:(45-50), ultrasonic extraction for 50-70min, filtering, collecting the filtrate, combining the filtrate, concentrating the obtained filtrate at a temperature of 85-95°C, filtering and collecting the filtrate for high-pressure sterilization, and obtaining the bletilla striata extract.

7. The hemostatic composition according to claim 1, characterized in that The preparation method of the Panax notoginseng extract comprises the following steps: crushing the Panax notoginseng raw material, adding 70%-75% ethanol, the solid-liquid ratio is 1:(12-15), reflux extraction at a temperature of 60-70°C for 40-60min, concentrating by freeze drying to obtain a concentrated extract, adding water to dissolve, filtering, adding 80% ethanol to the filtrate, and then concentrating by adsorption resin to obtain a second concentrated extract, then adding water and 90-95% ethanol to dissolve, then adding activated carbon to decolorize, and then filtering with a titanium rod to obtain a filtrate, refining and concentrating to obtain an extract, vacuum drying the extract, crushing it into fine powder, sieving, and obtaining the Panax notoginseng extract.

8. Use of the hemostatic composition according to any one of claims 1 to 7 in oral care products.

9. Use of the hemostatic composition according to claim 8 in oral care products, characterized in that: The mass content of the hemostatic composition in the oral care product is 0.6%-2.2%.

10. Use of the hemostatic composition according to claim 8 in an oral care product, wherein the oral care product comprises toothpaste, mouthwash or mouth spray.

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