CN117860958B

CN117860958B - Octopus-based ink-jet type temperature-sensitive antibacterial dressing, and preparation method and application thereof

Info

Publication number: CN117860958B
Application number: CN202410039388.XA
Authority: CN
Inventors: 朱舟; 乔铭薪; 万乾炳; 黄炜; 陈森林
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2024-01-10
Filing date: 2024-01-10
Publication date: 2024-09-03
Anticipated expiration: 2044-01-10
Also published as: CN117860958A

Abstract

The invention discloses an octopus-based ink-jet type temperature-sensitive antibacterial dressing, and a preparation method and application thereof, wherein the dressing comprises a shell and an antibacterial component filled in an inner cavity of the shell; the outer shell is of a box-shaped structure, the surface of the outer shell is provided with a plurality of nozzles, and the antibacterial components in the outer shell are released to the affected part through the nozzles. The dressing of the invention not only solves the problem that the wound dressing at the sport position is easy to fall off and deform, so that the wound dressing can be individually deformed according to the sport and keep the integrity of the material, but also realizes the intelligent controlled release of antibacterial components with inflammatory response through the temperature sensitivity of gelatin so as to promote wound healing, thereby providing a possible new scheme for solving the clinical problems. Meanwhile, the preparation method disclosed by the invention is simple and convenient to operate, low in cost and high in practicability, so that the dressing obtained by the preparation method has a good application prospect in wound healing of a sport part.

Description

Octopus-based ink-jet type temperature-sensitive antibacterial dressing, and preparation method and application thereof

Technical Field

The invention relates to the technical field of biomedical materials, in particular to an octopus-based ink-jet type temperature-sensitive antibacterial dressing, and a preparation method and application thereof.

Background

Skin is considered to be the most important barrier of the human body against invasion by physical, mechanical, chemical and pathogenic microorganisms, and when the integrity of the skin is compromised, bleeding, infection, pain, dysfunction, and even life threatening may result. Wound healing is a dynamic continuous process that includes hemostasis, inflammatory response, cell proliferation, angiogenesis and tissue regeneration, with corresponding changes in the wound microenvironment at different stages. The healing time of the wound is generally different in 3-14 days, the healing time of different parts is different, and the healing time of the wound of the jaw face is generally faster, and the wound of the part can be better recovered in 3-5 days because the blood supply of the jaw face is rich; the wound on the abdomen needs 5-7 days to heal, the wound on the chest and back needs 7-10 days, the wound on the limbs needs about 10-12 days, and the wound on the exercise parts such as joints is relatively longer in about 14 days. Wound healing is a natural and stable process under normal conditions, but when the organism immunity is low, the wound is exposed seriously, or the wounded suffers from systemic diseases such as diabetes mellitus, the invasion of pathogenic microorganisms easily occurs to cause wound infection, and the wound healing is hindered. To avoid this list of complications, we should perform bandaging interventions immediately after injury for vulnerable wounds.

In recent years, researches on wound healing dressings have been endless, such as wound dressings for diabetics, wound dressings with wet adhesion performance and the like, however, researches on wound healing dressings for maxillofacial and other sports parts have been relatively few, and although the wound blood supply of maxillofacial is short in healing time, maxillofacial wound healing is often affected by movements such as maxillofacial expression, chewing and the like, and in addition, wound healing at sports wounds such as joints at other parts of the body is greatly affected by movements. The wound at the exercise position is not only healed for a long time, but also the dressing is difficult to be completely attached to the wound due to exercise, so that the probability of wound infection is greatly increased. Therefore, the wound healing of the sports parts such as the maxillofacial part still has great challenges, so that a novel bionic sports wound dressing needs to be built, has enough ductility and toughness, and can realize complete fit and continuous antibacterial effect on the sports part wound, thereby promoting the healing of the sports part wound.

In view of this, the present invention has been made.

Disclosure of Invention

The invention aims to provide an octopus-based ink-jet type temperature-sensitive antibacterial dressing, a preparation method and application thereof.

The invention is realized in the following way:

in a first aspect, the invention provides an octopus-based ink-jet type temperature-sensitive antibacterial dressing, which comprises a shell and an inner antibacterial component filled in the inner cavity of the shell; the outer shell is of a box-shaped structure, the surface of the outer shell is provided with a plurality of nozzles, and the antibacterial components in the outer shell are released to the affected part through the nozzles.

In a second aspect, the invention also provides a preparation method of the octopus-based inkjet-type temperature-sensitive antibacterial dressing, which comprises the following steps: and printing the poloxamer 407 hydrogel solution out of a groove structure of the shell through a 3D printing technology, extruding mixed suspension of gelatin/zinc oxide composite hydrogel into the groove structure, and sealing the groove structure by using the poloxamer 407 hydrogel solution to form a box-shaped structure, wherein the sealed surface contains a plurality of nozzle structures.

In a third aspect, the invention also provides application of the octopus-based ink-jet type temperature-sensitive antibacterial dressing in preparation of medicines for promoting wound healing at a sport site.

The invention has the following beneficial effects:

The invention designs a novel composite dressing through a bionic structure of octopus ink-jet, the outer shell of the dressing is of a box-shaped structure, a plurality of nozzles are arranged on the surface of the box-shaped structure, and antibacterial components in the dressing are released through the nozzles. The dressing not only solves the problem that the wound dressing at the exercise is easy to fall off and deform, so that the wound dressing can be subjected to personalized deformation according to the exercise while keeping the integrity of the material, but also realizes the intelligent controlled release of antibacterial components with inflammatory response through the temperature sensitivity of gelatin so as to promote wound healing, and provides a possible new scheme for solving the clinical problem. Meanwhile, the preparation method disclosed by the invention is simple and convenient to operate, low in cost and high in practicability, so that the dressing obtained by the preparation method has a good application prospect in wound healing of a sport part.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Fig. 1 is a structural view of an octopus-based ink-jet type temperature-sensitive antibacterial dressing according to the present invention, wherein the upper view is an overall structure, and the lower view is a side view;

FIG. 2 is a graph showing the contact angle measurements of the outer ink sac and inner ink structure of the dressing of example 1;

FIG. 3 is a scanning electron microscope block diagram of the dressing of example 1;

FIG. 4 shows the comparison of cell proliferation activities of different experimental groups in experimental example 2;

FIG. 5 shows the results of cell adhesion experiments for different experimental groups in Experimental example 2;

FIG. 6 shows the results of bacterial plate coating for different experimental groups in Experimental example 3;

FIG. 7 is a scanning electron microscope image of E.coli and Staphylococcus aureus of different experimental groups in experimental example 3;

FIG. 8 is a transmission electron micrograph of E.coli and Staphylococcus aureus of different experimental groups in Experimental example 3.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

The inventor designs a temperature sensitive wound dressing which releases antibacterial components along with movement according to the principle of ink jet of an octopus ink sac, and the structure of the dressing is as follows:

the outer shell is in a box-shaped structure, the upper surface is provided with a plurality of nozzles; the antibacterial component of the inner cavity is in a gel state at normal temperature, and is filled in the inner cavity of the box-shaped structure. Wherein, the components of the shell comprise poloxamer 407 hydrogel, and the antibacterial components of the inner cavity comprise gelatin/zinc oxide composite hydrogel. The specific structure of the dressing is shown in fig. 1.

The dressing bionic octopus ink bag ink jet is designed with the structure, so that the dressing can not only meet the close fit of wound dressing at a sport position and not fall off along with continuous movement, but also continuously release ZnO along with movement to prevent wound infection, thereby promoting wound healing.

Specifically, poloxamer 407 (PO) corresponds to the ink sac structure of the octopus, gelatin/ZnO (GZ) mimics the ink structure in the octopus ink sac, and releases the relevant antibacterial factors along with the expansion and contraction of the outer ink sac structure during exercise.

In order to obtain better performance of the dressing, the inventor of the invention also optimizes the concentration of poloxamer 407, the concentration of gelatin and the concentration ratio of gelatin to zinc oxide so as to obtain better mechanical strength and toughness.

In the invention, the mass ratio of poloxamer 407 to water is 1:3-20, the mass ratio of gelatin to water is 1:5-20, and the mass ratio of gelatin to ZnO is 400-1000:1.

The poloxamer 407 is selected as the raw material of the shell, because the poloxamer 407 is a polyethylene glycol-polypropylene glycol-polyethylene glycol triblock copolymer, the invention has good thermal gelation property and excellent biocompatibility; the zinc oxide is selected as the antibacterial substance in the inner cavity because the ZnO nanomaterial can release Zn ²⁺, while Zn ²⁺ can react with-SH groups on proteins to destroy enzymes of an electron transfer system on a bacterial cell membrane. Meanwhile, znO is easier to generate active oxygen than other oxidized nano materials, so that ZnO has better antibacterial effects on staphylococcus aureus, escherichia coli and the like; gelatin is selected as a carrier because gelatin is a macromolecular hydrophilic collagen hydrolysate, is heated and dissolved into colloid, becomes gel-like when cooled to below 35-40 ℃, can become liquid when a wound is in inflammation heating condition, quickly releases ZnO, reduces skin surface temperature when inflammation does not occur or the inflammation subsides, deteriorates gelatin fluidity, and releases ZnO relatively slowly. From the results, poloxamer 407/gelatin/ZnO (PO/GZ) wound dressing realizes inflammatory response controlled release and slow release ZnO according to the temperature-sensitive change of gelatin.

The preparation method of the dressing with the structure comprises the following steps:

S1, adding poloxamer 407 powder into water, and uniformly stirring at the temperature of 2-4 ℃ to prepare poloxamer 407 hydrogel solution. Wherein the mass ratio of poloxamer 407 to water is 1:3-20.

S2, adding the gelatin powder into water, and uniformly stirring at 50-60 ℃ to prepare semitransparent uniform gelatin hydrogel solution. Wherein the mass ratio of the gelatin to the water is 1:5-20.

S3, adding ZnO powder into the gelatin hydrogel solution, and uniformly stirring at 50-60 ℃ to prepare suspension. Wherein the mass ratio of gelatin to ZnO is 400-1000:1.

S4, printing the poloxamer 407 hydrogel solution out of a groove structure of the shell through a 3D printing technology, extruding mixed suspension of gelatin/zinc oxide composite hydrogel into the groove structure, and sealing the groove structure through the poloxamer 407 hydrogel solution to form a box-shaped structure, wherein the surface of the seal is provided with a plurality of nozzle structures.

S5, obtaining the antibacterial temperature-sensitive dressing-PO/GZ.

The features and capabilities of the present invention are described in further detail below in connection with the examples.

Example 1

The embodiment provides an octopus-based ink-jet type temperature-sensitive antibacterial dressing, and the structure of the dressing is shown in fig. 1: the shell is poloxamer 407 hydrogel, the inner cavity is gelatin/zinc oxide composite hydrogel, and the specific preparation method is as follows:

(1) Adding poloxamer 407 powder into pure water according to the mass ratio of 1:3, and uniformly stirring at 2-4 ℃ to completely dissolve the poloxamer 407 powder into the pure water to form poloxamer 407 hydrogel solution;

(2) Adding gelatin powder into pure water according to the mass ratio of 1:5, heating to 50-60 ℃, and uniformly stirring to form semitransparent uniform gelatin hydrogel solution;

(3) Then adding ZnO powder into gelatin hydrogel solution according to the mass ratio of gelatin to ZnO of 400:1, heating to 50-60 ℃, and stirring to form uniform suspension;

(4) And printing the poloxamer 407 hydrogel solution out of a groove structure of the shell through a 3D printing technology, extruding mixed suspension of gelatin/zinc oxide composite hydrogel into the groove structure, and sealing the groove structure by using the poloxamer 407 hydrogel solution to form a box-shaped structure, wherein the sealed surface contains a plurality of nozzle structures.

(5) Finally, the antibacterial temperature-sensitive dressing-PO/GZ is obtained.

Example 2

The embodiment provides an octopus-based ink-jet type temperature-sensitive antibacterial dressing, wherein the shell of the dressing is poloxamer 407 hydrogel, the inner cavity is gelatin/zinc oxide composite hydrogel, and the specific preparation method comprises the following steps:

(1) Adding poloxamer 407 powder into pure water according to the mass ratio of 1:10, and uniformly stirring at 2-4 ℃ to completely dissolve the poloxamer 407 powder into the pure water to form poloxamer 407 hydrogel solution;

(2) Adding gelatin powder into pure water according to the mass ratio of 1:10, heating to 50-60 ℃, and uniformly stirring to form semitransparent and uniform gelatin hydrogel solution;

(3) Then adding ZnO powder into gelatin hydrogel solution according to the mass ratio of gelatin to ZnO of 800:1, heating to 50-60 ℃, and stirring to form uniform suspension;

Example 3

(1) Adding poloxamer 407 powder into pure water according to the mass ratio of 1:20, and uniformly stirring at 2-4 ℃ to completely dissolve the poloxamer 407 powder into the pure water to form poloxamer 407 hydrogel solution;

(2) Adding gelatin powder into pure water according to the mass ratio of 1:20, heating to 50-60 ℃, and uniformly stirring to form semitransparent uniform gelatin hydrogel solution;

(3) Then adding ZnO powder into gelatin hydrogel solution according to the mass ratio of gelatin to ZnO of 1000:1, heating to 50-60 ℃, and stirring to form uniform suspension;

Comparative example 1

The difference from example 1 is that the dressing was a dressing structure without gelatin/zinc oxide inner shell, namely poloxamer 407 (PO) dressing, and the comparative test results of comparative example 1 and experimental example 1 are as follows.

Experimental example 1

The dressing obtained in example 1 was subjected to contact angle detection by the following method: the relatively flat surfaces of PO and PO/GA were used as test surfaces, and the contact angle was calculated by measuring the angle between the contact line of the droplet (water) on the PO and PO/GA surfaces and the horizontal line. The number of PO and PO/GA groups was 6 in parallel.

The detection results are shown in fig. 2: the outer PO contact angle range was 19.179 ° and the inner GA contact angle range was 9.087 °.

The detection results prove that the dressing provided by the invention has good hydrophilicity, can release gelatin/ZnO (GZ) along with movement, can absorb inflammatory exudation at a wound, and can promote wound healing.

Meanwhile, the dressing obtained in the embodiment is observed through a scanning electron microscope, and the result is shown in fig. 3: the figure shows that the antibacterial auxiliary material has a good porous loose structure, and is beneficial to release and absorption of liquid.

Experimental example 2

The experimental example is a biological safety evaluation on the dressing in the example, and the evaluation method is as follows: first, cell proliferation-toxicity assays were performed for 1,3,5 days using the CCK-8 kit. Cells were seeded into 96-well plates and after 24 hours of incubation, media containing different sets of material was added to the plates. After 1d, 3d, 5d incubation, a pre-prepared CCK-8 solution was added to each well, incubated at 37 ℃ and absorbance measurements were performed using a microplate reader at a wavelength of 450nm, taking care to keep out light throughout the procedure. Then, in this experimental example, scanning electron microscope observation of cell adhesion was performed to further evaluate the biocompatibility, cells were inoculated onto the surface of the material, incubated for 4 hours, 1 day, and short-term and long-term adhesion morphology of the cells was observed by SEM. Cell samples were collected at different time points, cells were fixed using glutaraldehyde, and samples were dehydrated using gradient ethanol, and finally dried in a vacuum desiccator for SEM observation.

The experimental results are shown in fig. 4-5, and compared with the control CCK8 detection results, poloxamer 407 (PO) and poloxamer 407/gelatin/ZnO (PO/GZ) groups have no significant toxicity, and cells show good proliferation activity with increasing days. 4 hours and 1 day of cell adhesion experiments show that all 3 groups of cells have good adhesion and stretching morphology, and further prove that PO/GZ has good biocompatibility.

Experimental example 3

The experimental example is to verify the antibacterial performance of the dressing in the embodiment, and the verification method is as follows: coli (e.coil) and staphylococcus aureus (s.aureus) were used as subjects to examine the antibacterial ability of this example.

Firstly, an agar plate coating method is adopted to detect the antibacterial capability, PO or PO/GZ is used for treating bacterial suspension for 24 hours, and after dilution is carried out for a certain multiple, a coater is used for coating the dilution on an LB agar plate. The culture was performed at 37℃for 24 hours, and then the image was collected. Morphological changes of bacteria were observed using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM), sample preparation of SEM: and adding PO or PO/GZ into the bacterial suspension, incubating for 12 hours at 37 ℃, centrifugally collecting bacteria, fixing a sample by glutaraldehyde solution, sequentially dehydrating by gradient ethanol, drying the sample, spraying gold, and finally recording images by using a scanning electron microscope. Sample preparation of TEM: PO or PO/GZ is added to the bacterial suspension. After incubation for 12h at 37 ℃, bacteria are collected by centrifugation, glutaraldehyde and paraformaldehyde solution are added for fixation, then supernatant is removed by centrifugation, bacterial precipitate is collected at the bottom of a tube, soybean size is washed by PBS, supernatant is removed by centrifugation, new fixative is slowly added along the tube wall, then the tube is placed in a refrigerator at 4 ℃ for overnight, and finally TEM observation is carried out.

As shown in FIG. 6, the bacterial plate coating experiments showed that the PO/GZ group had good antibacterial effect against E.coli (E.coil) and Staphylococcus aureus (S.aureus), whereas the PO group did not have significant antibacterial effect compared to the control group.

As shown in fig. 7 and 8, the results of Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) also confirmed the conclusion that the e.coil and s.aureus of the PO group did not have significant shape and size changes compared to the control group. In contrast, the PO/GZ groups E.coil and S.aureus show shrinkage, wrinkles and partial cracks. Therefore, the PO/GZ antibacterial auxiliary material can be proved to have good antibacterial performance by the figures 6-8.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An octopus-based ink-jet type temperature-sensitive antibacterial dressing is characterized by comprising a shell and an antibacterial component filled in the inner cavity of the shell; the shell is of a box-shaped structure, the surface of the shell is provided with a plurality of nozzles, and the internal antibacterial components are released to an affected part through the nozzles;

the components of the shell comprise poloxamer 407 hydrogel, and the components of the inner cavity comprise gelatin/zinc oxide composite hydrogel.

2. The dressing of claim 1, wherein the contact angle of the poloxamer 407 hydrogel is 10-20 ° and the contact angle of the gelatin/zinc oxide composite hydrogel is 6-15 °.

3. A method of preparing an octopus inkjet-based temperature-sensitive antibacterial dressing according to claim 1 or 2, comprising: and printing the poloxamer 407 hydrogel solution out of a groove structure of the shell through a 3D printing technology, extruding a mixed suspension of gelatin/zinc oxide composite hydrogel into the groove structure, and sealing the groove structure by using the poloxamer 407 hydrogel solution to form a box-shaped structure, wherein the sealed surface contains a plurality of nozzle structures.

4. The method of preparing the poloxamer 407 hydrogel solution according to claim 3, comprising: adding poloxamer 407 powder into water, and uniformly stirring to prepare poloxamer 407 hydrogel solution; the temperature of the stirring is 2-4 ℃.

5. The preparation method according to claim 4, wherein the mass ratio of poloxamer 407 to water is 1:3-20.

6. The method of preparing the mixed suspension according to claim 5, wherein the method of preparing the mixed suspension comprises: adding the gelatin solution into water, and stirring uniformly at high temperature to prepare gelatin hydrogel solution; adding zinc oxide powder into gelatin hydrogel solution, stirring at high temperature to obtain suspension; the stirring temperature at the high temperature is 50-60 ℃.

7. The method of claim 6, wherein the mass ratio of gelatin to water is 1:5-20.

8. The method according to claim 7, wherein the mass ratio of gelatin to zinc oxide is 400-1000:1.

9. Use of an octopus inkjet-based temperature-sensitive antibacterial dressing according to claim 1 or 2 for the preparation of a medicament for promoting wound healing at a sport site.