CN110917391A - Polypeptide modified sodium alginate/PVA hydrogel dressing and preparation method thereof - Google Patents

Abstract

The invention relates to a polypeptide modified sodium alginate/PVA hydrogel dressing and a preparation method thereof, wherein the dressing comprises the following components in parts by weight: 1-3 parts of sodium alginate derivative, 6-9 parts of polyvinyl alcohol, 5-10 parts of glycerol, 0.1-5 parts of calcium chloride and 73-88 parts of purified water. The polypeptide modified sodium alginate/PVA double-network interpenetrating hydrogel dressing suitable for chronic wound care is prepared by blending the polypeptide modified sodium alginate and a PVA solution and combining a repeated freeze-thaw method and an ion crosslinking method, and has the functions of bioadhesive hemostasis, broad-spectrum antibiosis, wound exudate absorption, moisture retention and wound healing promotion.

Description

Polypeptide modified sodium alginate/PVA hydrogel dressing and preparation method thereof

Technical Field

The invention relates to the technical field of medical dressing preparation, in particular to a polypeptide modified sodium alginate/PVA hydrogel dressing and a preparation method thereof.

Background

With the continuous rise of the living standard of human beings and the development of aging population, the prevalence rate of diabetes at home and abroad is increased year by year, the diabetic foot is a common complication of a diabetic patient, a wound is difficult to heal and easy to infect, amputation or death can be caused seriously, great inconvenience and pain are brought to the life of the patient, and the development of the hydrogel wound dressing suitable for chronic wound care is an effective way for relieving the pain of the patient.

The hydrogel is a polymer which has a three-dimensional network structure and is insoluble and capable of swelling in water and retaining a large amount of water, and is formed by mutual crosslinking through covalent bonds, hydrogen bonds or van der waals forces. The hydrogel contains a large amount of water, is similar to human tissues, has excellent biocompatibility, and is widely applied to the field of medical dressings. The polyvinyl alcohol is safe and nontoxic, and in the process of forming gel, the hydrogen bond of hydroxyl groups among polyvinyl alcohol molecular chains can form a winding structure, so that the gel has excellent mechanical properties. The polyvinyl alcohol and the alginate are combined to prepare the hydrogel, so that the hydrogel has the effects of moisturizing and promoting wound healing. The natural polypeptide and the tissue have ionic electrostatic interaction and good tissue adhesion, and the biological adhesion of the gel can be enhanced and the broad-spectrum bacteriostasis effect can be achieved by introducing the natural polypeptide and the tissue into a gel system in a physical blending or grafting manner.

Disclosure of Invention

The invention aims to provide a polypeptide modified sodium alginate/PVA hydrogel dressing suitable for chronic wound care, which is applied to chronic wound care, more effectively prevents wound infection and promotes wound healing.

In order to solve the technical problems, the invention provides the following technical scheme: the paint comprises the following components in parts by weight: 1-3 parts of sodium alginate derivative, 6-9 parts of polyvinyl alcohol, 5-10 parts of glycerol, 0.1-5 parts of calcium chloride and 73-88 parts of purified water.

The preparation method of the polypeptide modified sodium alginate/PVA hydrogel dressing comprises the following steps:

s1: dispersing a certain amount of sodium alginate polypeptide derivative in glycerol, adding deionized water, and stirring for dissolving uniformly.

S2, preparing a polyvinyl alcohol aqueous solution, dissolving in a water bath at 85 ℃, mixing with S1, pouring into a mold, and repeatedly freezing and thawing for three times.

S3: and (4) soaking the gel obtained in the step (S2) in a calcium chloride solution, and then washing, drying, cutting, packaging and sterilizing to obtain the double-network interpenetrating sodium alginate polypeptide derivative/PVA hydrogel dressing.

The preparation method of the sodium alginate polypeptide derivative comprises the following steps:

s1: sodium alginate was dissolved in 2- (N-morpholino) ethanesulfonic acid (MES) buffer solution, and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide were added thereto and stirred at room temperature for 18 hours.

And S2, adding epsilon-polylysine, continuing stirring for 2 hours, dialyzing for 3 days, and freeze-drying to obtain the sodium alginate polypeptide derivative.

The invention has the beneficial effects that:

(1) the polypeptide modified sodium alginate/PVA hydrogel dressing dual-network interpenetrating gel dressing prepared by the method has stable performance and can be stored at normal temperature.

(2) The polypeptide modified sodium alginate/PVA hydrogel dressing has the advantages that the polypeptide is introduced through chemical grafting, the biological adhesiveness of the gel is enhanced, the bacterial growth is inhibited, the wound is adhered, and the hemostasis is accelerated.

(3) The alginate used as the raw material of the polypeptide modified sodium alginate/PVA hydrogel dressing has good biocompatibility, has the effect of promoting wound healing and has good biocompatibility.

(4) The polypeptide modified sodium alginate/PVA double-network interpenetrating hydrogel dressing suitable for chronic wound care is prepared by blending the polypeptide modified sodium alginate and a PVA solution and combining a repeated freeze-thaw method and an ion crosslinking method, and has the functions of bioadhesive hemostasis, broad-spectrum antibiosis, wound exudate absorption, moisture retention and wound healing promotion.

Drawings

FIG. 1 is a comparison graph of bacteriostatic test results.

Detailed Description

The first embodiment is as follows:

a preparation method of a polypeptide modified sodium alginate/PVA hydrogel dressing comprises the following steps:

s1: weighing 2.0g of sodium alginate polypeptide derivative, dispersing in 5mL of glycerol, adding 30mL of deionized water, and stirring uniformly to prepare a sodium alginate polypeptide derivative solution with a certain concentration.

S2, 8.0g of polyvinyl alcohol is weighed, 70mL of distilled water is added, and the mixture is dissolved uniformly in a water bath at 85 ℃. At room temperature, the sodium alginate polypeptide derivative solution and the polyvinyl alcohol solution are mixed evenly and poured into a mould. The mixture was then frozen in a refrigerator for 6 hours and thawed at room temperature for 2 hours, repeated three times.

S3: and (3) soaking the gel obtained in the step (S2) in a calcium chloride solution, and then washing, drying, cutting, packaging and completely killing to obtain the double-network interpenetrating sodium alginate polypeptide derivative/PVA hydrogel dressing.

Example two:

a preparation method of a polypeptide modified sodium alginate/PVA hydrogel dressing comprises the following steps:

s1: 3.0g of sodium alginate polypeptide derivative is weighed and dispersed in 5mL of glycerol, 30mL of deionized water is added and stirred uniformly, and sodium alginate polypeptide derivative solution with certain concentration is prepared.

S2, weighing 7.0g of polyvinyl alcohol, adding 70mL of distilled water, and dissolving uniformly in a water bath at 85 ℃. At room temperature, the sodium alginate polypeptide derivative solution and the polyvinyl alcohol solution are mixed evenly, poured into a mold, then placed into a refrigerator for freezing for 8 hours, unfrozen at room temperature for 3 hours, and repeated for three times.

S3: and (3) soaking the gel obtained in the step (S2) in a calcium chloride solution, and then washing, drying, cutting, packaging and completely killing to obtain the double-network interpenetrating sodium alginate polypeptide derivative/PVA hydrogel dressing.

Example three:

the preparation method of the sodium alginate polypeptide derivative comprises the following steps:

s1: preparing 100mL of 2- (N-morpholino) ethanesulfonic acid (MES) buffer solution with the concentration of 0.25mol/L, dropwise adding sodium chloride solution to adjust the pH value to 6.5, adding 1.0g of sodium alginate, and stirring until the sodium alginate is fully dissolved.

S2 adding 0.70g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and 0.26g of N-hydroxysuccinimide, stirring at normal temperature for 18 hours, adding epsilon-polylysine, continuing stirring for 2 hours, dialyzing with purified water for three days, and freeze-drying to obtain the sodium alginate polypeptide derivative.

Example four:

prepared by the invention

Sodium alginate derivatives and

the hydrogel dressings were separately subjected to in vitro cytotoxicity tests.

Reference is made to GB/T16886.5-2017 medical device biology evaluation part five: in vitro cytotoxicity assay methods.

Cell line: mouse fibroblast L929

Negative control group: 2% fetal bovine serum culture medium

Positive control group: 10% DMSO solution

Test groups: adding 2 percent fetal calf serum culture medium leaching liquor according to the proportion of 0.1g/ml of the sample mass, and shaking at the constant temperature of 37 ℃ for 24 +/-1 h.

Digesting cultured cells with vigorous growth for 48-72 h to obtain the cell with density of 1.5 × 10⁴Inoculating the cell suspension of each cell/ml into a 96-well plate, adding 100 mu L of the cell suspension into each well, and placing the cell suspension in 5% CO₂The culture was carried out in an incubator at 37 ℃ for 24 hours.

Discarding the original culture solution, adding 200 μ L negative control solution, positive control solution, and sample solution, placing 5 holes in each group, and placing in 5% CO₂The culture was carried out in an incubator at 37 ℃ for 48 hours. Adding 15 mu LMTT solution and 5% CO into each hole₂Culturing for 4h at 37 ℃ in an incubator, discarding the culture solution, adding 150 mu LDMSO respectively, shaking for 10min, detecting the absorbance at 490nm on an enzyme-labeling instrument, calculating the increment rate, and grading the in-vitro cytotoxicity reaction according to the table 1. The test results are shown in table 2.

TABLE 1 in vitro cytotoxicity response ranking

Toxicity rating (grade)	0	1	2	3	4
						Relative proliferation Rate (%)	≥100	80-99	50-79	30-49	0-29

TABLE 2 in vitro cytotoxicity test results

Sample (I)	2D
			Level 1
	Level 1
		Positive control	4 stage

As can be seen from Table 3, will

Sodium alginate derivatives and

the leaching liquor of the hydrogel dressing is used for culturing cells for 2 days respectively, and the cytotoxicity reaction grading is 1 grade, which shows that the sodium alginate derivative and the hydrogel dressing both have good biocompatibility.

Example five:

the polypeptide modified sodium alginate/PVA hydrogel dressing prepared by the invention is subjected to a bacteriostasis test.

The quantitative bacteriostasis test is carried out according to the carrier soaking method specified in WS/T650-2019.

Washing 24h fresh slant culture of test bacterium Escherichia coli with PBS, and diluting to 5.0 × 10⁶CFU/mL～5.0×10⁷CFU/mL bacterial suspension is ready for use. 10 mul of the bacterial suspension was applied by drop-dyeing to sterilized plain cloth pieces (10 mm. times.10 mm) and dried at 36 ℃. + -. 1 ℃ for use.

Weighing a sample according to the amount of 5 g/piece, placing the sample in a sterile plate, placing the sterile plate in a water bath at 20 +/-1 ℃ for 5min, taking a bacteria-staining cloth piece by using a sterile forceps, completely immersing the cloth piece in the sample, immediately timing, respectively taking out the bacteria-staining cloth piece, placing the bacteria-staining cloth piece into 5.0 mL PBS test tubes after 6 hours of interaction between a bacteria-staining carrier and the sample, uniformly mixing, oscillating, washing test bacteria, respectively absorbing 1.0 mL sample liquid, determining the number of viable bacteria according to a viable bacteria culture counting method, and inoculating 2 plates in each tube of sample liquid. The same batch of PBS was taken as negative control. All test samples and control samples were incubated at 36 ℃. + -. 1 ℃ for 48h to observe the results. The experiment was repeated 3 times, and the inhibition rate was calculated, and the experimental results are shown in fig. 1.

As can be seen from FIG. 1, compared with the blank group, the polypeptide-modified sodium alginate/PVA hydrogel dressing prepared by the invention has antibacterial activity, and the average antibacterial rate is 79.9%.

While specific embodiments of the present invention have been described above, the present invention is not limited to the above embodiments, and various modifications can be made within the scope of the technical gist of the present invention in the technical field of the present invention, as long as the general knowledge is grasped.

Claims (5)

1. The polypeptide modified sodium alginate/PVA hydrogel dressing is characterized by comprising the following components in parts by weight: 1-3 parts of sodium alginate derivative, 6-9 parts of polyvinyl alcohol, 5-10 parts of glycerol, 0.1-5 parts of calcium chloride and 73-88 parts of purified water.

2. The polypeptide-modified sodium alginate/PVA hydrogel dressing as claimed in claim 1, wherein the polypeptide-modified sodium alginate/PVA hydrogel dressing is prepared by mixing a sodium alginate polypeptide derivative with polyvinyl alcohol, and combining a repeated freeze-thaw method and an ion crosslinking method.

3. The polypeptide-modified sodium alginate/PVA hydrogel dressing as claimed in claim 2, wherein the sodium alginate polypeptide derivative is obtained by grafting sodium alginate with epsilon-polylysine.

4. The polypeptide-modified sodium alginate/PVA hydrogel dressing as claimed in claim 3, wherein the preparation method of the sodium alginate polypeptide derivative comprises the following steps:

s1: dissolving sodium alginate in 2- (N-morpholino) ethanesulfonic acid buffer solution, adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, and stirring at normal temperature for 18 hours;

and S2, adding epsilon-polylysine, continuing stirring for 2 hours, dialyzing for 3 days, and freeze-drying to obtain the sodium alginate polypeptide derivative.

5. The preparation method of the polypeptide-modified sodium alginate/PVA hydrogel dressing as claimed in any one of claims 1 to 4, characterized by comprising the following steps:

s1: dispersing a certain amount of sodium alginate polypeptide derivative in glycerol, adding deionized water, and stirring for dissolving uniformly;

s2, preparing a polyvinyl alcohol aqueous solution, dissolving in a water bath at 85 ℃, blending with S1, pouring into a mold, and repeatedly freezing and thawing for three times to obtain gel;

s3: and (4) soaking the gel obtained in the step (S2) in a calcium chloride solution, and then washing, drying, cutting, packaging and sterilizing to obtain the double-network interpenetrating sodium alginate polypeptide derivative/PVA hydrogel dressing.

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