CN115518005B

CN115518005B - Oral care gel and preparation method thereof

Info

Publication number: CN115518005B
Application number: CN202211304330.0A
Authority: CN
Inventors: 田亚光; 杨静文; 贾平一; 张赞; 蔡红宣
Original assignee: Hainan General Hospital
Current assignee: Hainan General Hospital
Priority date: 2022-10-24
Filing date: 2022-10-24
Publication date: 2023-06-20
Anticipated expiration: 2042-10-24
Also published as: CN115518005A

Abstract

The application relates to the technical field of oral care, and particularly discloses an oral care gel and a preparation method thereof. An oral care gel comprising the following raw materials in weight percent: 2-5% of morinda officinalis polysaccharide extract, 0.5-1% of gel and 1-3% of ornidazole; the preparation method comprises the following steps: mixing radix Morindae officinalis polysaccharide extract, gel, ornidazole, and water at a certain proportion, and packaging to obtain oral care gel. The oral care gel can be used for treating and preventing periodontitis, has a good anti-inflammatory effect, can reduce alveolar bone absorption, and can promote alveolar bone growth.

Description

Oral care gel and preparation method thereof

Technical Field

The present application relates to the technical field of oral care, and more particularly, to an oral care gel and a method of preparing the same.

Background

The oral cavity cleaning is irregular, dental plaque accumulated at the gingival and dental junction and harmful substances in the dental plaque act on the gingiva for a long time to cause inflammation, if the inflammation cannot be treated in time, the inflammation can be diffused from the gingiva to the deep layer to periodontal ligament, alveolar bone and cementum to develop into periodontitis, the periodontal ligament is destroyed due to the expansion of the inflammation, the alveolar bone is gradually absorbed, the gingiva is separated from the tooth root, so that a gingival sulcus is deepened to form a periodontal pocket, periodontal tissues are destroyed, and particularly when the absorption of the alveolar bone is aggravated, the phenomena of insufficient tooth supporting strength, loosening and displacement of teeth occur. Thus, oral care gels are particularly important.

At present, the oral care gel on the market mostly has the antibacterial and anti-inflammatory effects by adding antibiotics such as nitrozoles, imidazole derivatives, tetracyclines and the like, but the oral care gel on the market has no effects of reducing alveolar bone absorption and promoting alveolar bone regeneration, and even though periodontitis bacteria are killed, the injury of the alveolar bone cannot be repaired.

Disclosure of Invention

In order to reduce alveolar bone resorption and promote alveolar bone regeneration, the application provides an oral care gel and a preparation method thereof.

In a first aspect, the present application provides an oral care gel, employing the following technical scheme:

an oral care gel comprising the following raw materials in weight percent: 2-5% of morinda officinalis polysaccharide extract, 0.5-1% of gel and 1-3% of ornidazole.

By adopting the technical scheme, the morinda officinalis is prepared from the dried root of morinda officinalis of the madder family, has sweet, pungent and slightly warm nature, enters liver and kidney meridians, is one of four south-oriented medicines in China, and has the effects of resisting inflammation, easing pain, resisting tumor, resisting depression, enhancing immunity, resisting osteoporosis and the like.

The polysaccharide is one of important effective components of the morinda officinalis, and the morinda officinalis polysaccharide can exert an immunostimulation effect by inducing the expression of NO and inflammatory factors and down-regulating immunosuppressive cytokines in vitro; immune stimulation is induced in vivo by modulating the number of spleen immune cells, particularly increasing the number of Interferon-gamma (IFN-gamma) positive natural killer cells (Natural killer cells, NK), activating innate and adaptive immune responses. Especially in case of insufficient immune response, morinda citrifolia can exert a beneficial immunomodulating effect. However, under the condition of pathogenic microorganism infection, morinda officinalis inhibits the secretion of inflammatory factors such as TNF-alpha, IL-1 beta and the like by THP-1 macrophages induced by LPS, reduces inflammatory infiltration of an infection part, inhibits immune response through cytokines mediated by helper T cells Th1, th17, th22 and the like, restores immune balance and reduces tissue damage caused by excessive immune response. The morinda officinalis polysaccharide extract is added into the oral care gel, so that the oral care gel has the effects of preventing and treating chronic inflammation of the oral cavity and autoimmune alveolar bone absorption caused by dental plaque, has low-cost and easily-obtained raw materials, has no toxic or side effect, and is an ideal oral care gel of pure traditional Chinese medicine preparation.

Preferably, the weight percentage of the morinda citrifolia polysaccharide extract is 3-4%.

By adopting the technical scheme, the proportion of the morinda officinalis polysaccharide extract in the oral care gel is further optimized, and the performance of the oral care gel is optimized.

Preferably, the preparation method of the morinda officinalis polysaccharide extract comprises the following steps:

1) Pretreatment of

Reflux-extracting radix Morindae officinalis with ethanol for 2-3 times, filtering, and drying the residue;

2) Leaching with hot water

Reflux-extracting the residue obtained in step 1) with 80-90deg.C water for 2-3 times, filtering, centrifuging, and collecting supernatant to obtain radix Morindae officinalis polysaccharide crude extract;

3) Alcohol precipitation

Concentrating the polysaccharide crude extract obtained in the step 2) under reduced pressure to obtain a viscous extract, adding ethanol to make the ethanol content of the solution be 60-70%, mixing uniformly, precipitating with ethanol at 4-5deg.C for 12-14h, centrifuging, collecting precipitate, and freeze drying to obtain Morinda officinalis polysaccharide.

Through adopting above-mentioned technical scheme, carry out the preliminary treatment with alcohol extraction to the morinda officinalis, then carry out the leaching with hot water again, carry out alcohol precipitation again at last, alcohol extraction, hot water leaching, alcohol precipitation mutually support, improve the extraction rate of morinda officinalis polysaccharide.

Preferably, the ratio of the filter residue to the water in the step 2) is: 1g of filter residue corresponds to 10-15ml of water.

By adopting the technical scheme, the extraction rate of the morinda officinalis polysaccharide can be influenced by the feed liquid ratio of filter residues to water, the extraction rate of the morinda officinalis polysaccharide extract reaches the maximum level in the defined feed liquid ratio, and the extraction rate can be reduced due to the fact that the feed liquid ratio is too high or too low.

Preferably, the gel is sodium alginate hydrogel, and the preparation method comprises the following steps:

adding 10-15 parts by weight of sodium alginate into a sodium chloride solution, uniformly mixing, then treating for 30-40min at 45-50 ℃, and then standing for 10-12h to obtain a mixed solution; adding acrylamide into the mixed solution, uniformly mixing, adding 0.1-0.2 part by weight of ammonium persulfate, 0.003-0.005 part by weight of tetramethyl ethylenediamine and 0.09-0.15 part by weight of N, N-methylene bisacrylamide, and stirring at 0-3 ℃ for 15min; centrifuging and drying to obtain the sodium alginate hydrogel.

By adopting the technical scheme, the prepared sodium alginate gel has a porous structure and different gap sizes, and allows nutrient substances to diffuse and grow in cells so as to promote angiogenesis and osteogenesis in periodontal tissue regeneration, and good gaps can also accommodate drug particles to be inlaid therein to maintain drug release. In addition, the prepared sodium alginate gel can counteract the inhibition of ornidazole on cell growth, promote cell proliferation and promote alveolar bone regeneration.

Preferably, the preparation method of the sodium alginate hydrogel comprises the following steps:

adding 10-15 parts by weight of sodium alginate into a sodium chloride solution, uniformly mixing, then treating for 30-40min at 45-50 ℃, and then standing for 10-12h to obtain a mixed solution; adding acrylamide into the mixed solution, uniformly mixing, adding 0.1-0.2 part by weight of ammonium persulfate, 0.003-0.005 part by weight of tetramethyl ethylenediamine and 0.09-0.15 part by weight of N, N-methylene bisacrylamide, and stirring at 0-3 ℃ for 15min; then adding 0.1-0.2 weight parts of calcium peroxide particles, uniformly mixing, centrifuging and drying to obtain the sodium alginate hydrogel.

In the oral cavity, the vascular network is the only oxygen source of cells in the tissue, and the distance of oxygen diffusion in the tissue is short, when the oral gel is used for an oral injury affected part, the oxygen diffusion can be prevented, so that partial areas in the oral cavity are seriously anoxic, and the anoxic can cause cell death and tissue necrosis, thereby influencing the healing or regeneration of bone defects. By adopting the technical scheme, the calcium peroxide particles are added into the oral gel, and the calcium peroxide is one of the common solid oxygen release materials, and compared with hydrogen peroxide, the calcium peroxide has slow and durable oxygen release speed, so that the accumulation of local active oxygen is not easy to cause to destroy redox balance, the effective concentration of oxygen is maintained, the survival rate of stem cells is improved, the apoptosis induced by hypoxia is reduced, and the regeneration of alveolar bone is promoted.

Preferably, when the calcium peroxide particles are added, 0.15 to 0.3 parts by weight of ascorbic acid is added along with the calcium peroxide particles.

The calcium peroxide reacts with water to generate hydrogen peroxide and calcium hydroxide, wherein the hydrogen peroxide and the calcium hydroxide are strong oxidants, and the strong oxidants are continuously decomposed into oxygen and water; the latter is a strong base. Therefore, if the two are too much, the redox balance and acid-base balance of surrounding cells are destroyed, resulting in cell death and affecting bone regeneration. By adopting the technical scheme, the ascorbic acid is added so as to overcome the defect that excessive active oxygen and alkaline substances are generated in the process that the calcium peroxide releases oxygen when meeting water. Ensuring the normal growth of cells and promoting the regeneration of alveolar bone.

Preferably, the particle size of the calcium peroxide particles is 10-20nm.

The calcium peroxide particle size defined by the technical scheme ensures that the calcium peroxide particles are not easy to coagulate and the reaction speed is high; if the calcium peroxide particles are too large, the calcium peroxide particles are easy to coagulate into blocks when meeting water, and the reaction speed is too slow; if the calcium peroxide particles are too large, agglomeration is easy.

Preferably, it further comprises 0.4-1% fibronectin.

By adopting the technical scheme, the macromolecular extracellular membrane protein with fibronectin on the surfaces of various animal cells is the main non-collagenous glycoprotein in extracellular matrix and basement membrane, is evolutionarily conserved glycoprotein found in all tissues of human body, can serve as a three-dimensional bracket to guide the assembly of other extracellular matrix components, regulates cell behaviors through an integrin binding domain and a growth factor binding domain, promotes downstream reaction, promotes cell recruitment, proliferation and differentiation, and the fibronectin in oral gel can promote proliferation and regeneration of alveolar bone cells, thereby promoting regeneration of the alveolar bone.

In a second aspect, the present application provides a method for preparing an oral care gel, which adopts the following technical scheme:

a method of preparing an oral care gel comprising the steps of:

mixing radix Morindae officinalis polysaccharide extract, gel, ornidazole, and water at a certain proportion, and packaging to obtain oral care gel.

By adopting the technical scheme, the preparation method is simple and easy to operate, has no special requirement on production equipment, and is suitable for industrial production.

In summary, the present application has the following beneficial effects:

1. because the morinda polysaccharide extract is added into the oral gel, the inhibition rate of IL-6 inflammatory factor can reach 88.26-95.72%, and in the rat experiment, the new bone area can reach 0.378-0.407mm ² The alveolar ridge absorptivity can reach 13.83-20.87%, has good anti-inflammatory effect, reduces alveolar bone absorption, and promotes alveolar bone growth.

2. In the present application, calcium peroxide and ascorbic acid are preferably added to the gel, so that the promotion effect of the oral gel on alveolar bone regeneration is further improved.

Detailed Description

The present application is described in further detail below with reference to examples.

Preparation examples of starting materials and intermediates

Raw materials

The raw materials of the embodiment of the application can be obtained by the market:

the ethanol is 95% ethanol.

Preparation example

Preparation example I-1

A Morinda officinalis polysaccharide extract is prepared by the following steps:

1) Pretreatment of

Cleaning radix Morindae officinalis, oven drying, crushing, sieving with 40 mesh sieve, weighing 100g radix Morindae officinalis dry powder, reflux-extracting with 10 times volume of ethanol for 2 times, cooling, filtering, and drying the residue;

2) Leaching with hot water

Reflux-extracting the filter residue obtained in the step 1) with water at 90 ℃ for 2 times, wherein the ratio of the filter residue to the water is as follows: 1g of filter residue corresponds to 10ml of water, the extraction time is 2 hours, after the extraction is finished, the extraction is carried out, the filtration and the centrifugation are carried out, and the supernatant is taken to obtain the morinda officinalis polysaccharide crude extract;

3) Alcohol precipitation

Concentrating the polysaccharide crude extract obtained in the step 2) under reduced pressure to obtain a viscous extract, adding ethanol to ensure that the ethanol content of the solution is 65%, uniformly mixing, precipitating with ethanol at 4 ℃ for 14h, centrifuging, collecting precipitate, and freeze-drying to obtain the morinda officinalis polysaccharide.

PREPARATION EXAMPLE I-2

Unlike in preparation example 1, the number of times of hot water leaching in step 2) in preparation example 2 was 3.

Preparation example I-3

Unlike in preparation example 1, the ratio of the filter residue to water in step 2) in preparation example 3 is: 1g of filter residue corresponds to 15ml of water.

PREPARATION EXAMPLE I-4

Unlike in preparation example 1, the ratio of the filter residue to water in step 2) in preparation example 4 is: 1g of filter residue corresponds to 8ml of water.

PREPARATION EXAMPLE I-5

Unlike in preparation example 1, the ratio of the filter residue to water in step 2) in preparation example 5 is: 1g of filter residue corresponds to 20ml of water.

PREPARATION EXAMPLE I-6

1) Pretreatment of

Cleaning radix Morindae officinalis, oven drying, crushing, sieving with 40 mesh sieve, weighing 100g radix Morindae officinalis dry powder, reflux-extracting with 10 times volume of water for 2 times, cooling, filtering, and drying the residue;

2) Leaching with hot water

3) Alcohol precipitation

Preparation example II-1-II-3

A preparation method of the sodium alginate hydrogel comprises the following steps:

according to the raw material proportion in Table 1, sodium chloride is dissolved in water to prepare sodium chloride solution, sodium alginate is added into the sodium chloride solution for 3 times, stirring is continued for 4 hours at 25 ℃, stirring and mixing are carried out for 35 minutes at 45 ℃, and standing is carried out for 10 hours at 25 ℃ to obtain mixed solution; adding acrylamide into the mixed solution, uniformly mixing, adding ammonium persulfate, tetramethyl ethylenediamine and N, N-methylene bisacrylamide, and stirring at 2 ℃ for 15min; centrifuging and drying to obtain the sodium alginate hydrogel.

TABLE 1 preparation II-1-preparation II-3 raw materials proportioning table (g)

PREPARATION EXAMPLE II-4-II-6

Unlike preparation II-2, preparation II-4-II-6 also contains 0.1g, 0.2g and 0.3g of calcium peroxide particles, the particle size of which is 10-20nm, and the preparation method is as follows:

dissolving sodium chloride in water, preparing sodium chloride solution, adding sodium alginate into the sodium chloride solution for 3 times, continuously stirring for 4 hours at 25 ℃, stirring and mixing for 35 minutes at 45 ℃, and standing for 10 hours at 25 ℃ to obtain mixed solution; adding acrylamide into the mixed solution, uniformly mixing, adding ammonium persulfate, tetramethyl ethylenediamine and N, N-methylene bisacrylamide, and stirring at 2 ℃ for 15min; then adding calcium peroxide particles, uniformly mixing, and centrifuging and drying to obtain the sodium alginate hydrogel.

PREPARATION EXAMPLE II-7-II-9

In contrast to example II-5, examples II-7-II-9 also contained 0.15g, 0.30g, 0.45g of ascorbic acid, respectively, and anti-cyclohaemagglutinin was added together with the calcium peroxide particles.

Preparation example II-10

Unlike preparation II-8, the calcium peroxide particles in preparation II-10 had a particle diameter of 5 to 10nm.

Examples

Examples 1 to 5

An oral care gel, which is prepared by the following steps:

according to the raw material proportion table in Table 2, uniformly mixing the morinda officinalis polysaccharide extract, gel, ornidazole and water according to the proportion, and packaging to obtain oral care gel;

wherein the Morinda citrifolia polysaccharide extract is from preparation I-1 and the gel is from preparation II.

Table 2 examples 1-5 raw materials proportioning table (g)

Examples 8 to 12

Unlike example 7, the Morinda citrifolia polysaccharide extracts of examples 8-12 were derived from preparations I-2-I-6, respectively.

Examples 13 to 21

Unlike example 9, the Morinda citrifolia polysaccharide extracts of examples 13-21 were derived from preparations II-2-II-10, respectively.

Comparative example

Comparative example 1

Unlike example 1, comparative example 1 does not contain morinda citrifolia polysaccharide extract.

Comparative example 2

Unlike example 1, the weight percentage of morinda citrifolia polysaccharide extract in comparative example 2 was 10%.

Performance test

Detection method/test method

The following performance tests were performed on the oral care gels prepared in examples 1-21 and comparative examples 1-2, with the test results shown in Table 3: detecting the inhibition rate of IL-6 inflammatory factors by adopting a cell experiment of fibroblasts;

the method comprises the steps of taking rats as experimental objects, scanning and analyzing alveolar bones of each group of rats by adopting Micro-CT, performing Image processing and analysis by using Java (Image J-1.51r; NIH, USA) software, and measuring the new bone area and the alveolar ridge absorption rate, wherein the alveolar ridge absorption rate is calculated by the sum of the absorption rates of labial and lingual alveolar ridges.

TABLE 3 Performance test results

As can be seen from the combination of examples 1-21 and comparative examples 1-2 and the combination of table 3, the inhibition ratio of inflammatory factors in examples 1-21 is higher than that in comparative examples 1-2, which indicates that the oral gel of the present application has better effect in preventing and treating infectious oral inflammation; the alveolar ridge absorption rate in examples 1-21 was lower than that in comparative examples 1-2, indicating that the oral gel of the present application can reduce the absorption of alveolar bone by labial tongue; the area of new bone in examples 1-21 was higher than that of comparative examples 1-2, indicating that the oral gel of the present application can promote regeneration of alveolar bone.

As can be seen from the combination of example 1 and comparative examples 1 to 2 and the combination of table 3, the anti-inflammatory effect, the alveolar bone absorption reducing effect and the alveolar bone regeneration promoting effect of the oral gel of example 1 are superior to those of comparative examples 1 to 2, which are probably that the morinda polysaccharide extract has preventive and therapeutic effects on chronic inflammation of the oral cavity and autoimmune alveolar bone absorption caused by dental plaque through immunomodulation in the oral cavity, and the addition amount of the morinda polysaccharide extract is within the limit of the present application, and the effect is better.

It can be seen from the combination of examples 3 and examples 6 to 7 and the combination of Table 3 that the anti-inflammatory effect, the effect of reducing alveolar bone resorption, and the effect of promoting alveolar bone regeneration of the oral gel of examples 6 to 7 are superior to those of example 3, which is probably that the addition of fibronectin can enhance the regeneration of alveolar bone.

In combination with examples 3 and examples 8 to 12, and with Table 3, it can be seen that the oral gel of example 9 is superior in terms of inflammation inhibition and alveolar bone regeneration, which suggests that the Morinda citrifolia polysaccharide extract has a greater effect on inflammation inhibition and alveolar bone regeneration, whereas the process of extracting Morinda citrifolia polysaccharide extract of preparation I-3 is superior.

In combination with examples 13-17, and in combination with Table 3, it can be seen that the oral gels of examples 15-17 perform better in alveolar bone regeneration than examples 13-14, probably because calcium peroxide maintains the concentration of available oxygen in the oral cavity, improves stem cell survival, reduces hypoxia-induced apoptosis, and promotes alveolar bone regeneration.

In combination with examples 16-21 and with Table 3, it can be seen that the oral gel of examples 17-21 exhibited better alveolar bone regeneration than example 16, probably because the addition of ascorbic acid, which can overcome the deficiency of calcium peroxide addition, ensured normal growth of cells, and promoted alveolar bone regeneration.

The present embodiment is merely illustrative of the present application and is not intended to be limiting, and those skilled in the art, after having read the present specification, may make modifications to the present embodiment without creative contribution as required, but is protected by patent laws within the scope of the claims of the present application.

Claims

1. An oral care gel, characterized in that the gel comprises the following raw materials in percentage by weight: 2-5% of morinda officinalis polysaccharide extract, 0.5-1% of gel and 1-3% of ornidazole;

the gel is sodium alginate hydrogel, and the preparation method thereof comprises the following steps: adding 10-15 parts by weight of sodium alginate into a sodium chloride solution, uniformly mixing, then treating for 30-40min at 45-50 ℃, and then standing for 10-12h to obtain a mixed solution; adding 160-200 parts by weight of acrylamide into the mixed solution, uniformly mixing, adding 0.1-0.2 part by weight of ammonium persulfate, 0.003-0.005 part by weight of tetramethyl ethylenediamine and 0.09-0.15 part by weight of N, N-methylene bisacrylamide, and stirring at 0-3 ℃ for 15min; centrifuging and drying to obtain the sodium alginate hydrogel.

2. The oral care gel of claim 1, wherein: the weight percentage of the morinda officinalis polysaccharide extract is 3-4%.

3. The oral care gel of claim 1, wherein: the preparation method of the morinda officinalis polysaccharide extract comprises the following steps:

1) Pretreatment of

2) Leaching with hot water

3) Alcohol precipitation

4. An oral care gel according to claim 3, wherein: the ratio of the filter residue to the water in the step 2) is as follows: 1g of filter residue corresponds to 10-15ml of water.

5. The oral care gel of claim 1, wherein: the preparation method of the sodium alginate hydrogel comprises the following steps: adding 10-15 parts by weight of sodium alginate into a sodium chloride solution, uniformly mixing, then treating for 30-40min at 45-50 ℃, and then standing for 10-12h to obtain a mixed solution; adding 160-200 parts by weight of acrylamide into the mixed solution, uniformly mixing, adding 0.1-0.2 part by weight of ammonium persulfate, 0.003-0.005 part by weight of tetramethyl ethylenediamine and 0.09-0.15 part by weight of N, N-methylene bisacrylamide, and stirring at 0-3 ℃ for 15min; then adding 0.1-0.2 weight parts of calcium peroxide particles, uniformly mixing, centrifuging and drying to obtain the sodium alginate hydrogel.

6. The oral care gel of claim 5, wherein: when the calcium peroxide particles are added, 0.15 to 0.3 parts by weight of ascorbic acid is added along with the calcium peroxide particles.

7. The oral care gel of claim 5, wherein: the particle size of the calcium peroxide particles is 10-20nm.

8. The oral care gel of claim 1, wherein: it also comprises 0.4-1% fibronectin.

9. A method of preparing an oral care gel according to any one of claims 1 to 8, comprising the steps of: