CN109394701B - Biomedical material for treating gastric ulcer and preparation method thereof - Google Patents
Biomedical material for treating gastric ulcer and preparation method thereof Download PDFInfo
- Publication number
- CN109394701B CN109394701B CN201811513493.3A CN201811513493A CN109394701B CN 109394701 B CN109394701 B CN 109394701B CN 201811513493 A CN201811513493 A CN 201811513493A CN 109394701 B CN109394701 B CN 109394701B
- Authority
- CN
- China
- Prior art keywords
- gastric ulcer
- pollen
- biomedical material
- wall
- growth factor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1664—Compounds of unknown constitution, e.g. material from plants or animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/74—Synthetic polymeric materials
- A61K31/785—Polymers containing nitrogen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
- A61K33/10—Carbonates; Bicarbonates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/0065—Forms with gastric retention, e.g. floating on gastric juice, adhering to gastric mucosa, expanding to prevent passage through the pylorus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/167—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/167—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface
- A61K9/1676—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction with an outer layer or coating comprising drug; with chemically bound drugs or non-active substances on their surface having a drug-free core with discrete complete coating layer containing drug
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
Abstract
The invention relates to a biomedical material for treating gastric ulcer and a preparation method thereof, and relates to the field of medicines. The preparation method is obtained by loading calcium carbonate on the outer surface of the pollen wall to form a first sample and then coating polydopamine on the surface of the first sample, and the preparation method is simple and reliable to operate. The biomedical material for treating gastric ulcer provided by the invention can play the effects of resisting oxidation, resisting inflammation, neutralizing gastric acid and inhibiting ulcer enlargement at the wound forming place of gastric ulcer.
Description
Technical Field
The invention relates to the field of medicines, and in particular relates to a biomedical material for treating gastric ulcer and a preparation method thereof.
Background
Gastric ulcer is a common digestive tract disease, and mainly refers to tissue damage exceeding a muscularis mucosae layer caused by the self-digestion of gastric mucosa by gastric digestive fluid. The formation of gastric ulcers is related to the stress state of the body, the stimulation of physical and chemical factors, and the infection of certain pathogenic bacteria.
The existing gastric ulcer treatment method is mainly a chemical drug therapy and mainly has the problems that the chemical drug has large side effect on the stomach, has adverse reaction and long treatment period; the traditional Chinese medicine composition has more treatment methods for gastric ulcer, but the traditional Chinese medicine preparation has more varieties, great difference of treatment effect and unclear treatment mechanism.
Biomedical materials have been rapidly developed in recent years, and have been widely used in biological bodies for diagnosis, treatment, repair or replacement of damaged tissues and organs. However, biomedical materials applied to gastric ulcer are rare, and the invention is particularly proposed in view of the fact.
Disclosure of Invention
In view of the above problems, embodiments of the present invention provide a biomedical material for treating gastric ulcer, which can adhere to a mucous membrane and a ulcer surface, and can exert antioxidant, anti-inflammatory, gastric acid neutralizing, and ulcer enlargement inhibiting effects at a site where a wound of gastric ulcer is formed.
The embodiment of the invention provides a preparation method of a biomedical material for treating gastric ulcer, which is simple and reliable to operate, and the biomedical material for treating gastric ulcer, which can play the effects of resisting oxidation, resisting inflammation, neutralizing gastric acid and inhibiting ulcer enlargement at a wound forming place of the gastric ulcer, is obtained.
The technical problem to be solved by the invention is realized by adopting the following technical scheme.
The embodiment of the invention provides a preparation method of a biomedical material for treating gastric ulcer, which comprises the following steps:
calcium carbonate was loaded on the outer surface of the pollen wall to form a first sample.
And mixing the first sample with a weak alkaline buffer solution containing dopamine, and coating polydopamine on the surface of the first sample by utilizing self-polymerization of the dopamine.
The pollen wall is a natural pollen outer wall with a cavity structure, and is provided with a conveying channel communicated with the cavity.
The embodiment of the invention provides a biomedical material for treating gastric ulcer, which is prepared by the preparation method of the biomedical material for treating gastric ulcer.
The embodiment of the invention provides a biomedical material for treating gastric ulcer, which comprises a first sample and polydopamine coated on the surface of the first sample, wherein the first sample comprises a pollen wall with a cavity structure and calcium carbonate loaded on at least the outer surface of the pollen wall, the pollen wall is a natural pollen outer wall with a cavity structure, and the pollen wall is provided with a conveying channel communicated with the cavity.
The biomedical material for treating gastric ulcer and the preparation method thereof have the beneficial effects that:
the natural pollen outer wall with the adhesive capacity is used as a carrier, a layer of calcium carbonate is attached to the surface of the carrier to achieve the effects of neutralizing gastric acid and relieving gastric ulcer, and a layer of polydopamine is attached to the surface of the calcium carbonate and has the effects of resisting inflammation, resisting oxidation and adhering to mucosa, so that the effects of relieving and treating gastric ulcer are achieved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is an SEM image of an outer wall of sunflower spore powder provided in example 1 of the present invention;
FIG. 2 is an SEM image of a first sample provided in example 2 of the present invention;
FIG. 3 is an SEM photograph of biomedical materials for treating gastric ulcer provided in example 2 of the invention;
FIG. 4 shows the gavage of pDA @ CaCO after gastric ulcer caused by ethanol in mice provided in test example 2 of the present invention3Pictures of stomach HE staining after @ SEC 4 h;
FIG. 5 is a graph of HE in the stomach 14 days after administration of various drugs to the ethanol-induced gastric ulcer mice provided in test example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The biomedical material for treating gastric ulcer and the preparation method thereof according to the embodiment of the present invention will be described in detail below.
The invention provides a preparation method of a biomedical material for treating gastric ulcer, which comprises the following preparation steps:
s1, loading calcium carbonate on the outer surface of a pollen wall to form a first sample.
The pollen wall is a natural pollen outer wall with a cavity structure, the main component of the pollen outer wall is sporopollen, and the pollen outer wall has strong corrosion resistance and acid and alkali resistance, and is referred to as the pollen wall hereinafter. The pollen wall has a plurality of transport channels communicating with the cavity. The outer surface of the pollen outer wall is provided with bulges, such as semicircular, thorn-shaped or irregular bulges, and the like, namely the outer surface is a rough surface, so that calcium carbonate and polydopamine can be conveniently loaded at the later stage, and meanwhile, after gastric ulcer occurs, the surface structure of the gastric mucosa is damaged, a large number of inflammatory cells are present at the position where the gastric ulcer is formed, and the structure of the pollen outer wall can be adhered to the gastric mucosa, so that the targeted drug loading is achieved, and further targeted therapy is performed.
Wherein, in order to make the effect of treating gastric ulcer better, the stimulation to gastric ulcer or mucosa is less, and no burden is caused, optionally, the particle size of the pollen wall is 4-150 μm, for example, the particle size of the pollen wall is 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 70 μm, 90 μm, 100 μm, 120 μm or 140 μm, preferably 10-120 μm, which is convenient for operating and preparing the biomedical material for treating gastric ulcer.
In embodiments provided by the present invention, optionally, the pollen is selected from at least one of sunflower pollen, rape pollen, pollen pini and schisandra pollen, for example, the pollen is sunflower pollen, schisandra pollen or a mixture of rape pollen and pollen pini, etc. The above materials are readily available.
It should be noted that the pollen wall provided by the present invention can be purchased directly from the market, or can be prepared by itself, and optionally, in a preferred embodiment of this embodiment, the preparation method of the pollen wall includes: reacting the degreased pollen with acid or alkali at 50-80 deg.C and 300-900rpm for 5-20h, and washing with water, acetone, hydrochloric acid and ethanol for at least 1 time. Wherein, under the conditions of the temperature and the stirring speed, the inner wall of the pollen and the components such as protein can be sufficiently and effectively removed as long as the components are pectocellulose through acid or alkali reaction, and the substances are effectively removed through respectively cleaning by water, acetone, hydrochloric acid and ethanol, so as to obtain the pollen wall. In the present invention, the at least one time is, for example, one time, two times, three times, four times, five times, or the like.
Optionally, in order to reduce as much as possible the impurities contained in the obtained pollen wall, the sequence of washing is: sequentially cleaning with water, acetone, hydrochloric acid, water, acetone and ethanol at 40-60 deg.C for at least one time; that is, hot water, hot acetone, hot hydrochloric acid, hot water, hot acetone and hot ethanol are adopted to clean at least once in sequence. And after cleaning, drying to obtain a dry pollen wall for standby or direct use.
The defatted pollen may be purchased from the market directly or may be prepared by itself. The self-preparation adopts a mode of extracting by using ether, supercritical extraction and the like, for example, a mode of extracting by using ether, and specifically comprises the following steps: grinding dried pollen, adding diethyl ether as a solvent, and stirring at 600rpm at room temperature overnight, wherein the adding amount ratio of the pollen to the diethyl ether is 10-30 g: 170 ml, removing the ether by suction filtration after reaction, washing with the ether for a plurality of times, and drying at 50-70 ℃.
Because the pollen wall is acid-resistant and alkali-resistant, in a preferred embodiment of the invention, before the step of loading calcium carbonate on the outer surface of the pollen wall, the method further comprises the step of loading the effective components in the cavity through the delivery channel, so that the effective components are effectively prevented from being degraded in the transportation process, and the calcium carbonate can be released from the delivery channel after being decomposed in the stomach, i.e. the method has a slow release effect, so that the effective components are ensured to act on a target position for a long time, and the treatment effect is improved. It should be noted that the premise that the effective component is contained in the cavity through the conveying passage is that the particle size of the effective component needs to be smaller than the maximum conveying area of the conveying passage, so that the effective component can conveniently pass through the conveying passage and enter the cavity.
Wherein the method of encapsulating the active ingredient in the cavity via the passage comprises vacuum, high pressure, and turbine, and in a preferred embodiment of the invention, the method of encapsulating the active ingredient in the cavity via the delivery passage comprises:
mixing pollen wall and effective component solution uniformly, standing at 2-8 deg.C under vacuum condition of 10-20pa for 0.5-5 hr; the effective components can be effectively encapsulated in the cavity.
Wherein, optionally, the active ingredient is selected from at least one of a growth factor, a protein drug for treating gastric ulcer and a compound drug for treating gastric ulcer, wherein, it is to be noted that, an active ingredient which is not acid-resistant may also be within the selection range of the present invention.
The growth factor may be, for example, at least one of human basic fibroblast growth factor (bFGF), Keratinocyte Growth Factor (KGF), Epidermal Growth Factor (EGF), Nerve Growth Factor (NGF), dermal growth factor (AFGF), human fibroblast growth factor-21 (FGF-21), and human fibroblast growth factor-9 (FGF-9), and the protein for treating gastric ulcer and the compound for treating gastric ulcer are not specifically limited herein and may be purchased directly from the market.
In a preferred embodiment of the invention, the active ingredient is a growth factor, preferably a basic fibroblast growth factor; the basic fibroblast growth factor has the effects of effectively promoting healing and tissue repair of gastric mucosa injury and the like, promoting tissue regeneration and growth and development of nervous tissues and promoting treatment of gastric ulcer.
In order to further increase the loading of subsequent calcium carbonate and the like and enhance the purpose of relieving gastric ulcer, in a preferred embodiment of the invention, before the active ingredients are loaded in the cavity, pollen walls are dispersed in water and freeze-dried.
In a preferred embodiment of the invention, optionally, the loading capacity of the effective components is 1% -50% of the mass of the pollen wall; for example, the amount of the active ingredient loaded is 1%, 5%, 10%, 20%, 30%, 40%, or 50% of the pollen wall mass.
After gastric ulcer is formed, the structure of gastric mucosa is damaged, ulcer points appear, the gastric ulcer degree is aggravated due to the fact that the gastric acidity is about 1.2 at pH, therefore, calcium carbonate is loaded on the outer surface of the pollen wall, and the calcium carbonate is decomposed into Ca under the acidic condition2+And CO2The gastric ulcer medicament can relieve the damage of gastric acid to ulcer, and simultaneously calcium carbonate can neutralize gastric acid, so that the stomach can achieve a neutral environment, and the effective components can be protected to reach the gastric ulcer part, thereby achieving the treatment effect.
The step of loading calcium carbonate on the outer surface of the pollen wall can be that the pollen wall is placed in a soluble calcium salt solution and then added with a carbonate solution for reaction, or the pollen wall is placed in a carbonate solution and then added with a soluble calcium salt solution for reaction, in a preferred embodiment of the invention, the step of loading calcium carbonate on the outer surface of the pollen wall comprises the following steps: placing the pollen wall in a soluble calcium salt solution, and adding a carbonate solution for reaction; under the condition, Ca can be made2+Attached to pollen walls by adsorption of pollen wallsAnd the calcium carbonate is generated by the reaction on the outer surface of the pollen wall, and the obtained calcium carbonate is uniformly distributed on the outer surface of the pollen wall, so that the subsequent targeted therapy effect is good.
In a preferred embodiment of the present invention, the conditions for adding the carbonate solution include: adding carbonate solution under the conditions of 2-8 ℃, stirring speed of 200-500rpm and flow rate of 1-9 mL/h; under the condition, the obtained calcium carbonate can be generated in a particle form and attached to the surface of the pollen wall, and meanwhile, the calcium carbonate can be generated on the outer surface of the pollen wall more uniformly, so that the subsequent targeted therapy effect is good.
In a preferred embodiment of the present invention, the carbonate is at least one of sodium carbonate, ammonium bicarbonate and potassium carbonate; in a preferred embodiment of the invention, the soluble calcium salt is selected from at least one of calcium chloride and calcium nitrate.
In a preferred embodiment of the present invention, the loading amount of calcium carbonate in the first sample is 5% -20% of the mass of the first sample, such as 5%, 7%, 10%, 12%, 15%, 18% or 20%.
S2, mixing the first sample with a weak alkaline buffer solution containing dopamine, and wrapping the surface of the first sample with polydopamine by utilizing self-polymerization of the dopamine.
Dopamine is a compound with good biocompatibility and strong adhesion capacity, has the effects of oxidation resistance and inflammation resistance, can promote the repair of gastric ulcer, and can achieve double adhesion effects due to the adhesion performance of dopamine and the structural characteristics of pollen walls. The generation of polydopamine not only has the beneficial effect of dopamine, but also effectively protects the premature release of calcium carbonate and active ingredients in the process of conveying the biomedical material for treating gastric ulcer, and the effect of targeted quality cannot be achieved.
In a preferred embodiment of the present invention, the reaction conditions for self-polymerization of dopamine include:
and the poly-dopamine is reacted for 1 to 5 hours in a dark place at the temperature of between 1 and 8 ℃ and the speed of 100 and 500rpm, and under the conditions, the poly-dopamine has good reaction effect and complete package, thereby effectively preventing the premature release of calcium carbonate and active ingredients.
In a preferred embodiment of the invention, the weakly alkaline buffer is a Tris buffer with a pH value of 8-9, and the dopamine is added into the Tris buffer at a concentration of 1-5 mg/ml.
The invention also provides a biomedical material for treating gastric ulcer, which can be prepared by the preparation method of the biomedical material for treating gastric ulcer, and can also be prepared by other methods, but whatever method is adopted, the finally obtained biomedical material for treating gastric ulcer structurally comprises a first sample and polydopamine coated on the surface of the first sample, the first sample comprises a natural pollen wall with a cavity structure and calcium carbonate loaded on at least the outer surface of the pollen wall, the pollen wall is a natural pollen outer wall with a cavity structure, the pollen wall is provided with a conveying channel communicated with the cavity, and the number of the conveying channels is one or at least two, which is not limited herein.
In the invention, the natural pollen extract with the adhesive capacity is used as a medicine or protein carrier, a layer of calcium carbonate is attached to the surface of the carrier to neutralize gastric acid and relieve gastric ulcer, and a layer of polydopamine is attached to the surface of the calcium carbonate and has the functions of anti-inflammation, antioxidation and adhesion to mucosa, so that various effects of treating gastric ulcer are achieved.
In a preferred embodiment of the present invention, the pharmaceutical composition further comprises an active ingredient entrapped in the cavity, the active ingredient being at least one selected from the group consisting of a growth factor, a protein drug for treating gastric ulcer, and a compound drug for treating gastric ulcer.
That is, aiming at the phenomenon that protein drugs or acid intolerant drugs are easy to inactivate in the gastric acid environment, the protein drugs or the acid intolerant drugs and other drugs for treating gastric ulcer are matched with pollen walls, calcium carbonate and polyacrylic acid, so that the effect of treating gastric ulcer in various modes is further achieved, and the biomedical materials for treating gastric ulcer need to be stored at low temperature of 2-7 ℃ subsequently.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
Extraction of sunflower spore powder outer wall (SEC)
1.1 extraction of defatted sunflower pollen
Taking 20g of dried sunflower pollen, adding 200ml of diethyl ether into a round-bottom flask, stirring at 600rpm at room temperature overnight, removing the diethyl ether by suction filtration, washing with diethyl ether for three times, drying at 60 ℃, and storing at room temperature.
1.2 extraction of sunflower spore powder outer wall (SEC)
Taking 10g of degreased sunflower pollen, grinding, adding 150mL of 85% phosphoric acid, reacting for 10h at 70 ℃ and 600rpm, washing with 200mL of hot water for 3 times, washing with 100mL of hot acetone for two times, washing with 100mL of 2M hot HCl for 2 times, washing with 200mL of hot water for 3 times, washing with 100mL of hot acetone for two times, washing with 100mL of hot ethanol for two times, obtaining yellow solid particles, drying at 60 ℃, obtaining brown SEC, wherein an electron microscope scanning image of the SEC is shown in figure 1.
As can be seen from figure 1, the SEC surface is covered with the thorn-shaped structure, and in nature, the thorn-shaped structure has strong adhesive capacity and can be adhered to the legs or the body surface of the insect to achieve the aim of pollination. After gastric ulcer occurs, the surface structure of gastric mucosa is damaged, a large number of inflammatory cells appear at the position where the gastric ulcer is formed, and the SEC structure can be adhered to the gastric mucosa to achieve the effect of targeted drug loading.
Example 2
A preparation method of a biomedical material for treating gastric ulcer comprises the following steps:
1. preparation of the first sample
2mg of SEC prepared in example 1 were taken and concentrated in 0.2mL of 0.1M CaCl2Dispersing uniformly, then injecting 0.2mL Na with the concentration of 0.05M by using a syringe pump2CO3Reacting for 1min under the reaction condition of 150rpm, 4 ℃ and the injection speed of 0.5mL/min, centrifuging to remove supernatant at 12000rpm for 4min after the reaction is finished, then washing for 1 time by using 0.2mL deionized water, collecting solid, and freeze-drying to obtain a light brown first sample.
Fig. 2 is a scanning electron microscope image of the first sample, and it can be observed that a layer of calcium carbonate is adhered to the SEC surface, and the calcium carbonate can achieve the effects of neutralizing gastric acid, relieving gastric ulcer, maintaining bFGF activity, and achieving the effect of treating protein drugs.
2. Preparation of biomedical material for treating gastric ulcer
Dissolving 4mg of dopamine in 2mL of tris-buffer solution with the pH value of 8.4, adding 2mg of the first sample, uniformly dispersing, and reacting for 3 hours in a dark place at the temperature of 4 ℃ and the stirring speed of 100 rmp. After the reaction is finished, centrifuging for 4min at 1200rmp, removing supernatant, washing for 2 times by using 0.2mL deionized water, and freeze-drying to obtain the grey-black biomedical material for treating gastric ulcer.
Fig. 3 is a scanning electron microscope image of the biomedical material for treating gastric ulcer, which shows that polydopamine is adhered to the surface of the biomedical material for treating gastric ulcer, the pollen thorn-shaped structure is shortened, and pDA on the surface of the biomedical material for treating gastric ulcer has the effects of oxidation resistance, inflammation resistance and adhesion pair, can promote the adhesion of the material to the gastric surface, and achieves the effects of targeted drug delivery and gastric ulcer repair promotion.
Example 3
A preparation method of a biomedical material for treating gastric ulcer comprises the following steps:
1. preparation of SEC carrying bFGF in the Cavity
5mg of SEC prepared in example 1 were dispersed in 2ml of water and lyophilized after freezing at-20 ℃. And then adding 0.1mL of bFGF solution with the concentration of 5mg/mL into the obtained SEC, performing ultrasonic treatment to uniformly mix the solution, performing vacuum treatment at 4 ℃ for 2h in a vacuum drying oven with 10-20Pa to enable the bFGF to enter an SEC cavity structure through a conveying channel, centrifuging at 12000rpm for 4min after the vacuum treatment to remove a supernatant, washing 1 time with 0.1mL of deionized water, collecting precipitates, and freeze-drying to obtain the bFGF-loaded SEC in the cavity.
2. Preparation of the first sample
Take 2mg of SEC with bFGF in the cavity and put it in 0.2mL of 0.1M CaCl2Dispersing uniformly, then injecting 0.2mL Na with the concentration of 0.05M by using a syringe pump2CO3Reacting for 1min under the reaction condition of 150rpm, 4 ℃ and the injection speed of 0.2mL/min, centrifuging to remove supernatant after the reaction is finished at 12000rmp for 4min, then washing for 1 time by using 0.2mL deionized water, collecting light brown solid, and freeze-drying to obtain a first sample.
3. Preparation of biomedical material for treating gastric ulcer
Dissolving 4mg of dopamine in 2mL of tris-buffer solution with the pH value of 8.4, adding 2mg of the first sample, uniformly dispersing, and reacting for 3 hours in a dark place at the temperature of 4 ℃ and the stirring speed of 100 rmp. After the reaction is finished, centrifuging for 4min at 1200rmp, removing supernatant, washing for 2 times by using 0.2mL deionized water, and freeze-drying to obtain the grey-black biomedical material for treating gastric ulcer.
Example 4
A preparation method of a biomedical material for treating gastric ulcer comprises the following steps:
1. preparation of SEC carrying bFGF in the Cavity
4mg of SEC from example 1 were dispersed homogeneously in water by adding 2ml of water, frozen at-20 ℃ and freeze-dried. And then adding 0.12mL of bFGF solution with the concentration of 5.5mg/mL into the obtained SEC, performing ultrasonic treatment to uniformly mix the solution, performing vacuum treatment in a 10Pa vacuum drying oven at 4 ℃ for 2.5h to ensure that the bFGF enters an SEC cavity structure through a conveying channel, centrifuging at 12000rpm for 3.5min after the vacuum treatment to remove a supernatant, washing with 0.1mL of deionized water for 1 time, collecting precipitates, and freeze-drying to obtain the bFGF-coated SEC in the cavity.
2. Preparation of the first sample
Take 2mg of SEC with bFGF in the cavity and put it in 0.2mL of 0.1M CaCl2Dispersing uniformly, then injecting 0.2mL Na with the concentration of 0.05M by using a syringe pump2CO3Reacting for 2min under the reaction condition of 160rpm, 5 ℃ and the injection speed of 0.7mL/min, centrifuging for 4min at 12000rmp after the reaction is finished, removing supernatant, washing for 1 time by using 0.2mL deionized water, collecting solid, and freeze-drying to obtain a first sample.
3. Preparation of biomedical material for treating gastric ulcer
Dissolving 3.8mg of dopamine in 2mL of tris-buffer solution with the pH value of 8.4, adding 2.1mg of the first sample, uniformly dispersing, and reacting for 3 hours in a dark place at the temperature of 4.5 ℃ and the stirring speed of 100 rmp. After the reaction is finished, centrifuging for 4min at 1200rmp, removing supernatant, washing for 2 times by using 0.2mL deionized water, and freeze-drying to obtain the product.
Example 5
The difference between the embodiment and the embodiment 3 is only in the step 1, 5mg of the SEC prepared in the embodiment 1 is directly added into 0.1mL of bFGF solution with the concentration of 5mg/mL, the solution is mixed evenly by ultrasonic, the bFGF enters a SEC cavity structure in a vacuum drying oven with 10-20Pa at 4 ℃ under vacuum for 2h, after the vacuum is finished, 12000rpm is carried out, supernatant is centrifuged for 4min, 0.1mL of deionized water is used for washing for 1 time, and the precipitate is collected and then lyophilized to obtain brown SEC with the bFGF encapsulated in the cavity.
Example 6
The difference between this example and example 1 is only that pollen adopted is pollen pini.
Test example 1
The preparation method of the biomedical material for treating gastric ulcer prepared in example 2 is used for obtaining the SEC prepared in example 1 and the first sample (CaCO used in the invention) prepared in example 2 respectively through EDS (EDS) spectrogram3@ SEC) and the biomedical material for the treatment of gastric ulcers prepared in example 2 (in the present invention pDA @ CaCO)3@ SEC) and the results are shown in table 1.
TABLE 1 elemental contents
| Element content | C | O | Ca |
| SEC | 38.9 | 61.1 | - |
| CaCO3@SEC | 27.9 | 64.93 | 7.66 |
| pDA@CaCO3@SEC | 33.26 | 60.74 | 6.01 |
From Table 1, it was found that the Ca content of the first sample was reduced after the surface of the first sample was attached with pDA, and it was confirmed that pDA @ CaCO3@ SEC construction was successful.
Meanwhile, the results of the tests of examples 3 to 5, respectively, show that the calcium content in the first sample is the highest, and the content of Ca element is reduced after pDA is attached, which indicates that the structures of the biomedical materials for treating gastric ulcer are constructed successfully. Meanwhile, through comparison analysis of EDS (electron Desorption spectroscopy) energy spectrum charts, the content of the calcium element loaded in example 3 is higher than that of the calcium element loaded in example 5, which indicates that the freeze-drying step is beneficial to the loading of calcium carbonate.
Test example 2
1. Intragastric administration of pDA @ CaCO after gastric ulcer of mice caused by ethanol3Test of @ SEC
Starving 5 baby/c mice 20-30g for about 4 months for 24h under the condition of sufficient water source, and then performing intragastric administration of 0.2ml of 75% ethanol to form an acute gastric ulcer model. 2h mice were gavaged with 10mg/ml pDA @ CaCO prepared in example 23@ 0.2ml SEC, taking out the stomach after 4h, taking out the stomach after the mouse is killed by removing the neck, fixing the stomach tissue by 4% paraformaldehyde, gradually dehydrating by alcohol, embedding paraffin, and slicing the tissue to the thickness of about 5 μm. The tissue sections were stained with HE staining kit and the staining results are shown in fig. 4.
The results are shown in FIG. 4. Wherein FIGS. 4A and 4B are pDA @ CaCO administered after gastric ulcer caused by ethanol in mice3After @ SECHE staining pictures of different parts can be seen from the arrow positions in fig. 4A and 4B, the HE staining can obviously show that biomedical material particles for treating gastric ulcer are deposited in the gastric ulcer area and are adhered to the gastric ulcer area, and the material is proved to have good adhesion effect and can be enriched in the wound area, so that a better drug delivery effect is achieved.
2. Therapeutic effects of different drugs on gastric ulcer mice
30 babl/c mice 20-30g for about 4 months are randomly divided into 5 groups, and starved for 24 hours under the condition of sufficient water source. Randomly selecting 4 groups of hungry mice, and forming a gastric ulcer model by intragastric administration of 0.2ml of 75% ethanol; one group was given 0.2ml of physiological saline as a normal control group. The normal control group mice were gavaged 2h later with 0.2ml of physiological saline. After 2 hours in 4 groups of gastric ulcer model mice, one group was administered with 0.2ml of physiological saline by gavage, one group was administered with 0.4mg/ml of bFGF by gavage, and one group was administered with 10mg/ml of pDA @ CaCO prepared in example 2 by gavage3@ SEC 0.2ml, one group was given 10mg/ml pDA @ CaCO by gavage3@SEC@bFGF(pDA@CaCO3@ SEC @ bFGF biomedical Material for the treatment of gastric ulcers, prepared in example 3, per mg pDA @ CaCO3@ SEC @ bFGF contains about 40ug of bFGF) 0.2 ml. The stomach is taken after the mice are removed from the neck and die for 14 days, the stomach tissue is fixed by 4 percent paraformaldehyde, and is dehydrated step by alcohol, embedded by paraffin, sliced by tissue, and the thickness is about 5 mu m. The tissue sections were stained with HE staining kit and the staining results are shown in fig. 5.
FIG. 5 is a graph of HE in the stomach 14 days after administration of various drugs to ethanol-induced gastric ulcer mice. In FIG. 5, (a) is an HE image of a normal mouse stomach; (b) stomach HE images after administration of physiological saline to gastric ulcer mice; (c) gastric HE images after bFGF administration to gastric ulcer mice; (d) administration of pDA @ CaCO to gastric ulcer mice3@ SEC post gastric HE image; (e) administration of pDA @ CaCO to gastric ulcer mice3The HE image of gastric tissue after @ SEC @ bFGF.
As is apparent from FIG. 5, the stomach was still in the reparative phase after the administration of physiological saline and bFGF to the gastric ulcer mice, with large cavities (arrows) between cells of the gastric tissue, while the gastric ulcer mice were administered pDA @ CaCO3After @ SEC, there were few cavities between the stomach tissues and the cells were well-aligned, and the gastric ulcer mice were given pDA @ CaCO3After @ SEC @ bFGF, gastric tissue is well repaired, almost no hollow holes are formed in the tissue, the tissue cell rows are compact, and the curative effect is good.
The pDA @ CaCO provided in example 4 and example 5 was used3@ SEC @ bFGF instead of pDA @ CaCO as provided in example 33@ SEC @ bFGF, tested as described above, gave almost identical experimental results: the gastric tissue is well repaired, and almost no cavity is formed in the tissue and the tissue cell rows are compact. Through the tests, the biomedical material for treating the gastric ulcer provided by the embodiment of the invention is expected to have the effects of relieving and treating the gastric ulcer for a human body.
In summary, the biomedical material for treating gastric ulcer provided by the embodiment of the invention takes the natural pollen wall with adhesive capacity as a carrier, a layer of calcium carbonate is attached to the surface of the carrier to achieve the effects of neutralizing gastric acid and relieving gastric ulcer, and then a layer of polydopamine is attached to the surface of the calcium carbonate, and the polydopamine has the effects of anti-inflammation, antioxidation and mucosa adhesion, so that the effects of relieving and treating gastric ulcer are achieved. The preparation method of the biomedical material for treating the gastric ulcer is simple and reliable, and the biomedical material for treating the gastric ulcer, which can play the effects of resisting oxidation, resisting inflammation, neutralizing gastric acid and inhibiting ulcer enlargement at the wound forming place of the gastric ulcer, is obtained.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (19)
1. A preparation method of a biomedical material for treating gastric ulcer is characterized by comprising the following steps:
loading calcium carbonate on the outer surface of the pollen wall to form a first sample;
mixing the first sample with a weak alkaline buffer solution containing dopamine, and coating the surface of the first sample with polydopamine by utilizing the self-polymerization of the dopamine;
the pollen wall is a natural pollen outer wall with a cavity structure, and is provided with a conveying channel communicated with the cavity;
before the step of loading the calcium carbonate on the outer surface of the pollen wall, the preparation method further comprises loading an active ingredient selected from at least one of a growth factor, a protein drug for treating gastric ulcer and a compound drug for treating gastric ulcer into the cavity through the delivery passage;
the growth factor is at least one selected from human basic fibroblast growth factor, keratinocyte growth factor, epidermal growth factor and nerve growth factor.
2. The method for preparing a biomedical material for treating gastric ulcer according to claim 1, wherein the active ingredient is basic fibroblast growth factor.
3. The method for preparing a biomedical material for treating gastric ulcer according to claim 1, wherein the method for encapsulating the effective ingredient in the cavity through the delivery passage comprises:
mixing the pollen wall and the solution of the effective components uniformly, and standing at 2-8 deg.C under vacuum condition of 10-20Pa for 0.5-5 h.
4. The method for preparing the biomedical material for treating the gastric ulcer as claimed in claim 1, wherein the loading amount of the effective components is 1% -50% of the mass of the pollen wall.
5. The method of claim 1, wherein the step of loading the effective component into the cavity further comprises dispersing the pollen wall in water and lyophilizing.
6. The method for preparing a biomedical material for treating gastric ulcer according to claim 1, wherein the step of loading the calcium carbonate on the outer surface of the pollen wall comprises:
and (3) placing the pollen wall in a soluble calcium salt solution, and adding a carbonate solution for reaction.
7. The method for preparing a biomedical material for the treatment of gastric ulcer according to claim 6, characterized in that the conditions of adding said carbonate solution comprise: the carbonate solution was added at 2-8 deg.C, a stirring rate of 200 and 500rpm, and a flow rate of 1-9 mL/h.
8. The method for preparing biomedical material for treating gastric ulcer according to claim 6, characterized in that said carbonate is at least one of sodium carbonate, ammonium bicarbonate and potassium carbonate.
9. The method for preparing a biomedical material for the treatment of gastric ulcer according to claim 6, characterized in that said soluble calcium salt is selected from at least one of calcium chloride and calcium nitrate.
10. The method for preparing biomedical materials for the treatment of gastric ulcer according to claim 1, wherein the loading amount of calcium carbonate in the first sample is 5% -10% of the mass of the first sample.
11. The method for preparing biomedical material for treating gastric ulcer according to claim 1, wherein the reaction conditions of self-polymerization of dopamine comprise:
the reaction is carried out for 1 to 5 hours at the temperature of 1 to 8 ℃ and under the condition of 100 and 500rpm protected from light.
12. The method for preparing biomedical material for treating gastric ulcer according to claim 1, wherein said weakly alkaline buffer is Tris buffer with pH value of 8-9, and the concentration of dopamine added in said Tris buffer is 1-5 mg/ml.
13. The method for preparing a biomedical material for the treatment of gastric ulcer according to any one of claims 1 to 12, characterized in that said pollen wall is prepared by a method comprising:
the method comprises the steps of reacting the degreased pollen for 5-20 hours by acid or alkali at the temperature of 50-80 ℃ and under the condition of 300-900rpm, and then respectively cleaning the pollen for at least 1 time by water, acetone, hydrochloric acid and ethanol.
14. The process for the preparation of biomedical material for the treatment of gastric ulcers according to claim 13, characterized in that said washing is carried out in the sequence: sequentially cleaning the raw materials by water, acetone, hydrochloric acid, water, acetone and ethanol at 40-60 ℃.
15. The method for preparing biomedical material for the treatment of gastric ulcer according to claim 13, wherein said pollen wall has a particle size of 4-150 μm.
16. The method for preparing biomedical material for the treatment of gastric ulcer according to claim 13, wherein said pollen wall has a particle size of 10-120 μm.
17. The method for preparing biomedical material for treating gastric ulcer as claimed in claim 13, wherein said pollen is at least one selected from sunflower pollen, rape pollen, pine pollen and schisandra pollen.
18. A biomedical material for treating gastric ulcer, which is prepared by the preparation method of the biomedical material for treating gastric ulcer according to any one of claims 1 to 17.
19. A biomedical material for treating gastric ulcer is characterized by comprising an effective component, a first sample and polydopamine, wherein the polydopamine is coated on the surface of the first sample, the first sample comprises a natural pollen wall with a cavity structure and calcium carbonate loaded at least on the outer surface of the pollen wall, the pollen wall is a natural pollen outer wall with a cavity structure, the pollen wall is provided with a conveying channel communicated with the cavity, the effective component is loaded in the cavity, and the effective component is selected from at least one of a growth factor, a protein drug for treating gastric ulcer and a compound drug for treating gastric ulcer;
wherein the growth factor is at least one selected from human basic fibroblast growth factor, keratinocyte growth factor, epidermal growth factor and nerve growth factor.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811353121 | 2018-11-14 | ||
| CN2018113531219 | 2018-11-14 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109394701A CN109394701A (en) | 2019-03-01 |
| CN109394701B true CN109394701B (en) | 2021-04-06 |
Family
ID=65458578
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811513493.3A Active CN109394701B (en) | 2018-11-14 | 2018-12-11 | Biomedical material for treating gastric ulcer and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109394701B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110104610B (en) * | 2019-04-10 | 2022-10-18 | 康元大工生物技术(大连)有限公司 | Pollen structure particle and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966441A (en) * | 2010-09-09 | 2011-02-09 | 天津大学 | Polydopamine microcapsules and preparation method thereof |
| RU2011106811A (en) * | 2011-02-24 | 2012-08-27 | Общество с ограниченной ответственностью "ДИНАМИК ДЕВЕЛОПМЕНТ ЛАБОРАТОРИЗ" (RU) | LIQUID COMPOSITION INCREASING PERFORMANCE AND POSSESSING ADAPTOGENIC, WELL-HEALING, ANTI-INFLAMMATORY ACTION, AND METHOD FOR PRODUCING IT |
| US20140105961A1 (en) * | 2012-10-15 | 2014-04-17 | Texas Tech University System | Immunomodulation using spores and pollen grains |
| CN107668033A (en) * | 2017-09-12 | 2018-02-09 | 国家纳米科学中心 | A kind of farm chemical carrier based on calcium carbonate and preparation method thereof |
| CN108434119A (en) * | 2018-03-07 | 2018-08-24 | 华中农业大学 | A kind of albumen takes orally the preparation method of microcapsule formulation |
| CN108836950A (en) * | 2018-06-12 | 2018-11-20 | 华中农业大学 | A kind of albumen sporopollenin takes orally the preparation method of microcapsules |
-
2018
- 2018-12-11 CN CN201811513493.3A patent/CN109394701B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966441A (en) * | 2010-09-09 | 2011-02-09 | 天津大学 | Polydopamine microcapsules and preparation method thereof |
| RU2011106811A (en) * | 2011-02-24 | 2012-08-27 | Общество с ограниченной ответственностью "ДИНАМИК ДЕВЕЛОПМЕНТ ЛАБОРАТОРИЗ" (RU) | LIQUID COMPOSITION INCREASING PERFORMANCE AND POSSESSING ADAPTOGENIC, WELL-HEALING, ANTI-INFLAMMATORY ACTION, AND METHOD FOR PRODUCING IT |
| US20140105961A1 (en) * | 2012-10-15 | 2014-04-17 | Texas Tech University System | Immunomodulation using spores and pollen grains |
| CN107668033A (en) * | 2017-09-12 | 2018-02-09 | 国家纳米科学中心 | A kind of farm chemical carrier based on calcium carbonate and preparation method thereof |
| CN108434119A (en) * | 2018-03-07 | 2018-08-24 | 华中农业大学 | A kind of albumen takes orally the preparation method of microcapsule formulation |
| CN108836950A (en) * | 2018-06-12 | 2018-11-20 | 华中农业大学 | A kind of albumen sporopollenin takes orally the preparation method of microcapsules |
Non-Patent Citations (3)
| Title |
|---|
| Extraction of sporopollenin exine capsules from sunflower pollen grains;Raghavendra C. Mundargi et al;《RSC ADVANCES》;20161231;第6卷(第20期);第16533-16539页 * |
| Interactions of mussel-inspired polymeric nanoparticles with gastricmucin: Implications for gastro-retentive drug delivery;Suhair Sunoqrot et al;《Colloids and Surfaces B: Biointerfaces》;20170503;第156卷;第1-8页 * |
| 抗消化性溃疡药物作用的新靶点-生长因子;柳丽等;《药学进展》;20040430;第28卷(第4期);第159-162页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109394701A (en) | 2019-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dhamecha et al. | Applications of alginate microspheres in therapeutics delivery and cell culture: Past, present and future | |
| Chi et al. | Chinese herb microneedle patch for wound healing | |
| US20190192654A1 (en) | Microparticulated Vaccines for the Oral or Nasal Vaccination and Boostering of Animals Including Fish | |
| TWI445555B (en) | Dressing comprising active components of centella asiatica and use of the same | |
| CN108837184A (en) | A kind of composite membrane and preparation method thereof for Guided Bone Regeneration | |
| KR20100015294A (en) | Solid film, rapidly dissolvable in liquids | |
| CN109394701B (en) | Biomedical material for treating gastric ulcer and preparation method thereof | |
| CN1197552C (en) | Preparations for the application of anti-inflammatory agents | |
| CN107427561A (en) | Skin wound therapeutic combination | |
| CN101229162A (en) | Freeze-dried powder injection containing dextrogyrate rabeprazole and salts thereof, preparing technology thereof | |
| CN101744815B (en) | Composite medicament of omeprazole sodium | |
| CN100388921C (en) | Medicine for promoting healing of wound | |
| CN109432219A (en) | A kind of thiosugar aluminium porcelain enamelling | |
| Liu et al. | Poly (lactic-co-glycolic acid)-Chitosan–gelatin composite nanomaterials for the treatment of diabetic foot ulcer wound infection | |
| CN101947220A (en) | Combined medicament of omeprazole sodium | |
| JP5716214B2 (en) | Bread seed-based mixture | |
| CN114732782A (en) | Needle tip liquid, microneedle patch, and preparation method and application thereof | |
| CN109568645A (en) | A kind of composite growth factor promotees to repair gel and the preparation method and application thereof | |
| CN108096199B (en) | Omeprazole sodium for injection and preparation method thereof | |
| CN113967192A (en) | Pharmaceutical composition for accelerating wound healing, preparation method and application thereof | |
| CN102670528A (en) | Pantoprazole sodium lyophilized powder composition for injection and preparation method thereof | |
| CN108721688A (en) | A kind of aerogel dressing preparation method containing honeysuckle, wolfberry fruit extract | |
| CN1257705A (en) | Flowable drug precursor product for compressing tablet, pill and coated tablet, and preparing method thereof | |
| CN101491518B (en) | Combination for treating tumor | |
| CN104434992A (en) | Biological adhesive vaginal tablet of periplaneta americana extract and preparation method of biological adhesive vaginal tablet |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |