1. A probe for controlling release of hydrogen sulfide in response to wound exudate is characterized in that: is more than one of formula I or formula II;

in the formula I and the formula II, R is phenylene, thienylene, furylene, pyrrolylene or phosphole without two hydrogens; r₁、R₂Is independent hydrogen, C_1-6Alkyl radical, C_1-6Alkyloxy, C_1-6An alkylthio group.

2. The probe for controlled release of hydrogen sulfide in response to wound exudate according to claim 1, characterized in that:wherein the wavy line represents the attachment site; x is O, S, NH or PH.

3. The probe for controlled release of hydrogen sulfide in response to wound exudate as claimed in claim 2, wherein: x is S; the probe is more than one of a formula I-1 or a formula II-1;

4. the probe for controlled release of hydrogen sulfide in response to wound exudate as claimed in claim 3, wherein: r₁、R₂Is a separate hydrogen, and is,

at the moment, the probe is more than one of a formula I-2 or a formula II-2;

5. the method for preparing the probe for the response and controlled release of hydrogen sulfide from wound exudate according to any one of claims 1 to 4, which is characterized in that: the method comprises the following steps:

1) dissolving a precursor compound in an organic solvent to obtain a precursor compound solution; will contain S^2-Or HS^-Preparing the compound into a solution to obtain a sulfur-containing solution;

2) mixing the precursor compound solution with a sulfur-containing solution to obtain a probe;

the precursor compound has the structure of formula III:

r is phenylene, thienylene, furanylene, pyrrolylene, phospha-cyclopentadiene deprived of two hydrogens; r₁、R₂Is independent hydrogen, C_1-6Alkyl radical, C_1-6Alkyloxy, C_1-6An alkylthio group;

said containing S^2-Or HS^-The compound of (A) is a solvent-soluble S-containing compound^2-Or HS^-A salt;

the volume ratio of the organic solvent to the sulfur-containing solution is (60-98) to (40-2).

6. The method for preparing a probe for controlling release of hydrogen sulfide in response to wound exudate according to claim 5, wherein the probe comprises:wherein the wavy line represents the attachment site; x is O, S, NH, PH;

said containing S^2-Or HS^-The compound of (A) is a water-soluble S-containing compound^2-Or HS^-A salt; the sulfur-containing solution is to contain S^2-Or HS^-Dissolving the compound of (a) in a solvent to obtain a sulfur-containing solution;

the volume ratio of the organic solvent to the sulfur-containing solution is (90-98) to (10-2); the total volume of the organic solvent and the sulfur-containing solution is 100 parts;

the probe triggers the release of hydrogen sulfide by water or aqueous solutions.

7. The method for preparing a probe for controlling release of hydrogen sulfide in response to wound exudate according to claim 6, wherein the probe comprises:

r isX is S; at this time, the precursor compound has the structure of formula III-1:

said containing S^2-Or HS^-The compound of (A) is a water-soluble S-containing compound^2-Or HS^-A salt comprising: more than one of sodium sulfide, sodium hydrosulfide, potassium sulfide, potassium hydrosulfide and magnesium sulfide.

8. The method for preparing a probe for controlling release of hydrogen sulfide in response to wound exudate according to claim 5, wherein the probe comprises:

the precursor compound and the compound containing S^2-The mol ratio of the compound (1-2.5) to 1; the precursor compound and the compound containing HS^-The mol ratio of the compound (A) is (0.5-1.5) to 1; said containing S^2-Or HS^-The compound of (A) is a compound containing S^2-And/or containing HS^-A compound of (1);

the organic solvent is an organic solvent capable of dissolving the precursor compound; more specifically one or more of dimethyl sulfoxide, methanol, ethanol, acetonitrile and tetrahydrofuran.

9. The use of a probe for controlled release of hydrogen sulfide in response to wound exudate according to any one of claims 1 to 4, wherein: the probe for the wound exudate response controlled release hydrogen sulfide is used for preparing wound repair materials or preparations.

10. An ion-responsive material, characterized by: comprising a precursor compound having the structure of formula III:

r is phenylene, thienylene, furanylene, pyrrolylene, phospha-cyclopentadiene deprived of two hydrogens; r₁、R₂Is independent hydrogen, C_1-6Alkyl radical, C_1-6Alkyloxy, C_1-6An alkylthio group;

the ion is S^2-Or HS^-。

Background

The skin is in direct contact with the external environment, is the largest organ of the human body, is also the largest and most active immune organ, and is the first defense barrier of the human body. The skin can protect various internal tissues from pathogens, mechanical damage, and extreme temperatures. In addition, the skin plays a vital role in thermoregulation, sensory perception, vitamin D synthesis and protection from external damage (including uv light, trauma and microorganisms). Since the skin covers the outermost layers of the human body, the skin is very susceptible to various types of injuries, such as wounds, cuts, burns, chemical injuries, ulcers and cancers, which have a significant impact on both the patient and the health care economy. Skin wound repair is a very complex physiological process requiring the sequential, intricate and complex regulation of many different types of cells, chemokines, cytokines and growth factors. The healing of the wound surface comprises four successive stages, namely hemostasis, inflammation, proliferation and remodeling.

Hydrogen sulfide (H)₂S) is the third endogenous gas transmitter found following Nitric Oxide (NO) and carbon monoxide (CO). Hydrogen sulfide has a variety of biological functions in the cardiovascular, nervous, reproductive, and endocrine systems. In addition, the hydrogen sulfide can also promote the repair of skin wounds by inhibiting oxidative stress, relieving inflammation, activating vascular endothelial factors, promoting angiogenesis and other mechanisms. Therefore, the hydrogen sulfide donor is used as a prodrug for repairing skin wounds, can replace oral hydrogen sulfide medicines, and provides targeted local treatment for wound repair. However, the existing materials are difficult to realize real-time self-monitoring of hydrogen sulfide release, and toxic byproducts can be generated in the release process.

Wound repair based on hydrogen sulfide requires that the drug delivery system have certain characteristics, such as controlled release, appropriate physicochemical properties, good storage stability, real-time self-monitoring and advanced delivery systems. Therefore, in order to realize real-time self-monitoring of hydrogen sulfide release, it is necessary to develop a hydrogen sulfide donor probe that is easy to prepare and has good biocompatibility, and to construct a novel hydrogen sulfide release system.

Disclosure of Invention

In order to overcome the defects and shortcomings of the prior art, the invention aims to provide a probe for controlling the release of hydrogen sulfide in response to wound exudate and a preparation method thereof.

Another object of the present invention is to provide an application of the above probe. The probe is used for preparing wound repair materials or preparations, in particular to wound repair materials of exudate response controlled release hydrogen sulfide.

The purpose of the invention is realized by the following technical scheme:

a probe for wound exudate response controlled release hydrogen sulfide is more than one of formula I or formula II;

in the formula I and the formula II, R is phenylene, thiophenylene (a group losing two hydrogens on thiophene), furanylene, pyrrolylene or phosphole losing two hydrogens; r₁、R₂Is independent hydrogen, C_1-6Alkyl radical, C_1-6Alkyloxy, C_1-6An alkylthio group;

preferably, the first and second electrodes are formed of a metal,wherein the wavy line represents the attachment site; x is O, S, NH, PH, more preferably X is S.

The probe is preferably more than one of formula I-1 or formula II-1;

R₁、R₂preferably hydrogen alone.

At the moment, the probe is more than one of a formula I-2 or a formula II-2;

the preparation method of the probe for the wound exudate response controlled release hydrogen sulfide comprises the following steps:

1) dissolving a precursor compound in an organic solvent to obtain a precursor compound solution; will contain S^2-Or HS^-Preparing the compound into a solution to obtain a sulfur-containing solution;

2) mixing the precursor compound solution with a sulfur-containing solution to obtain a probe;

the precursor compound has the structure of formula III:

in the formula III, R₁，R₂As defined above for formula I, formula II; in particular, R is phenylene, thienylene (a group on a thiophene which loses two hydrogens), furanylene, pyrrolylene, phosphacyclopentadiene which loses two hydrogens; r₁、R₂Is independent hydrogen, C_1-6Alkyl radical, C_1-6Alkyloxy, C_1-6An alkylthio group;

preferably, the first and second electrodes are formed of a metal,wherein the wavy line represents the attachment site; x is O, S, NH, PH; more preferably R isX is S.

When X is S, the precursor compound has the structure of formula III-1:

further, R₁、R₂When hydrogen is independent, the precursor compound has the structure of formula III-2:

said containing S^2-Or HS^-The compound of (A) is a solvent-soluble S-containing compound^2-Or HS^-Salt, preferably soluble in water, containing S^2-Or HS^-Salts, such as: sodium sulfide, sodium hydrosulfide, potassium sulfide, potassium hydrosulfide, magnesium sulfide, etc.

The sulfur-containing solution is to contain S^2-Or HS^-Dissolving the compound of (a) in a solvent to obtain a sulfur-containing solution; the solvent is water.

The volume ratio of the organic solvent to the sulfur-containing solution is (60-98) to (40-2); preferably (90-98): (10-2). The total volume of the organic solvent and the sulfur-containing solution is 100 parts.

The precursor compound and the compound containing S^2-The mol ratio of the compound (1-2.5) to 1; the precursor compound and the compound containing HS^-The molar ratio of the compound (A) is (0.5-1.5) to 1. Said containing S^2-Or HS^-The compound of (A) is a compound containing S^2-And/or containing HS^-The compound of (1).

The organic solvent is an organic solvent capable of dissolving the precursor compound; preferably at least one of dimethyl sulfoxide, methanol, ethanol, acetonitrile and tetrahydrofuran.

The probe of the invention triggers the release of hydrogen sulfide by water or aqueous solutions; when the release of hydrogen sulfide is triggered, the volume ratio of water in the whole system is more than or equal to 70 percent, and preferably more than or equal to 90 percent.

The probe is used for preparing wound repair materials or preparations, in particular to wound repair materials of exudate response controlled release hydrogen sulfide. The probe of the invention is used as a drug for wound repair.

The precursor compound is used for preparing ion-responsive materials, and the ion is S^2-And/or HS^-。

Compared with the prior art, the invention has the following advantages and beneficial effects:

1. the probe can realize quick and efficient hydrogen sulfide release and real-time self-monitoring under water response (the probe realizes self-monitoring through the change of the color of the probe).

2. The probe of the invention can promote the repair of the wound surface and has good repair effect.

Drawings

In FIG. 1, (A) shows that precursor compound III-2 is added with Na of different concentrations in solution₂(iv) ultraviolet-visible absorption spectroscopy after S; (B) is accompanied by Na₂The S concentration is increased, and the line graph of the absorption relative value of III-2 at 355nm and 580nm is plotted; the inset shows that III-2 is added with 0.5 equivalent of Na₂Bright field photos before and after S;

FIG. 2 (A) shows the UV-visible absorption spectra of precursor compound III-2 after different concentrations of NaHS were added to the solution; (B) is a plot of the relative values of the absorbance at 355nm versus 580nm for III-2 as the NaHS concentration increases; the inset is a bright field photograph of III-2 before and after adding 1 equivalent of NaHS to the solution;

FIG. 3 (A) shows an ultraviolet-visible light absorption spectrum of a precursor compound III-2 and a probe I-2 or II-2 in DMSO or water; (B) is a bright field photograph of III-2(1, 4), I-2(2, 5) or II-2(3, 6) in DMSO or water;

FIG. 4 (A) is a standard curve of absorbance versus hydrogen sulfide concentration; (B) is the concentration of hydrogen sulfide released by probe I-2 or II-2 triggered by water;

FIG. 5 shows different concentrations of Na₂Cell viability of L929 cells 24 hours after S or precursor compound III-2 treatment;

FIG. 6 is a diagram showing the healing process of the wound surface in the blank group, the control group and the probe I-2 group (0.5mmol/L group and 1mmol/L group);

FIG. 7 is a bar graph of the rate of healing of wounds at various time points for the blank, control, probe I-2 groups (0.5mmol/L and 1mmol/L groups);

FIG. 8 is a representative H & E staining of the wound surface of the blank group, the control group, and the probe I-2 group (0.5mmol/L group and 1mmol/L group) at each time point, (B) is an enlarged view of the corresponding region in (A), and the yellow line is the boundary between the normal tissue and the wound surface.

Detailed Description

The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.

Preparation of precursor Compound III-2:

5- (4- (diphenylamine) phenyl) thiophene-2-carbaldehyde (354mg, 1.0mmol) and 2- (3-cyano-4, 5, 5-trimethylfuran-2 (5H) -methylene) malononitrile (301mg, 1.5mmol) were placed in an ethanol/tetrahydrofuran mixed solvent (volume ratio 6: 4), and the reaction was stirred under reflux under a nitrogen atmosphere for 5 hours. After the reaction is finished, purifying by a silica gel column chromatography method to obtain III-2 with the yield of 59 percent.¹H NMR(CDCl₃，400MHz)：δ7.81(d，J＝16.0Hz，1H)，8.20(dt，J₁＝8.8Hz，J₂＝4.4Hz，J₃＝2.4Hz，2H)，7.44(d，J＝4.4Hz，1H)，7.34(d，J＝1.6Hz，1H)，7.33-7.28(m，4H)，7.19-7.09(m，6H)，7.06(d，J＝8.8Hz，2H)，6.62(d，J＝15.6Hz，1H)，1.76(s，6H)；¹³C NMR(CDCl₃，101MHz)：δ175.65，173.06，153.94，149.56，146.67，139.61，137.89，137.47，129.57，127.19，125.48，124.26，124.06，121.89，112.08，112.00，111.30，110.87，104.97，97.05，96.71，77.33，77.22，77.01，76.69，29.70，26.54.ESI-HRMS：m/z[M+H⁺]Theory C₃₄H₂₅N₄OS⁺537.1749; actually, 537.1735.

III-2：

Example 1

Preparation of Probe I-2:

weighing 10mg of precursor compound III-2, dissolving with DMSO to prepare a solution of 10.5 mmol/L; weighing 5mg of Na₂S, dissolving with ultrapure water to prepare a solution of 100 mmol/L; 950. mu.L of 10.5mmol/L solution of III-2 was added with 50. mu.L of Na₂And mixing the solution of S in 100mmol/L to obtain the probe I-2.

The structure of I-2 is as follows:

example 2

Preparation of Probe II-2:

weighing 10mg of precursor compound III-2, dissolving with DMSO to prepare a solution of 10.5 mmol/L; weighing 5mg of NaHS, dissolving with ultrapure water and preparing into 200mmol/L solution; taking 950 mu L of 10.5mmol/L solution of III-2, adding 50 mu L of 200mmol/L solution of NaHS, and mixing uniformly to obtain the probe II-2.

The structure of II-2 is:

example 3

Precursor Compounds III-2 vs S^2-Response of (2):

adding Na with ultrapure water₂S is prepared into 1.0mmol/L mother liquor, and different volumes of the mother liquor are added into a DMSO/water mixed solution containing the precursor compound III-2. After the solutions were mixed well and incubated at room temperature for 0.5 hours, the uv-vis absorption spectra of the mixed solutions were tested on a uv spectrophotometer to investigate the precursor compound III-2 vs S^2-The response activity of (2). The final concentration of the precursor compound III-2 was 10.0. mu. mol/L, S^2-Gradually increased from 0.0. mu. mol/L to 5.0. mu. mol/L.

In FIG. 1, (A) shows that precursor compound III-2 is added with Na of different concentrations in solution₂(iv) ultraviolet-visible absorption spectroscopy after S; (B) is accompanied by Na₂The S concentration is increased, and the line graph of the absorption relative value of III-2 at 355nm and 580nm is plotted; the inset shows that III-2 is added with 0.5 equivalent of Na₂Bright field photographs before and after S.

As can be seen from FIG. 1, with S^2-The precursor compound III-2 gradually decreases in the characteristic absorption peak at about 580nm with increasing concentration, and simultaneously increases in the absorption value at about 355nm to form a new absorption peak, and the relative absorption values at 355nm and 580nm tend to increase. When 0.5 equivalent of S is added^2-Then, the absorption peak of precursor compound III-2 at around 580nm almost completely disappeared. Furthermore, with S^2-The color of the solution gradually becomes lighter from the original blue color and finally becomes colorlessThe change in color of the solution is also indicative of the precursor compounds III-2 and S^2-The addition process of (1). The results show that the precursor compound III-2 can react with S^2-And (4) high-efficiency addition.

Example 4

Precursor compound III-2 to HS^-Response of (2):

NaHS was dissolved in ultrapure water and prepared as a 1.0mmol/L mother liquor, and the mother liquor was added to a DMSO/water mixed solution containing precursor compound III-2 in various volumes. After the solutions were mixed well and incubated at room temperature for 0.5 hours, the uv-vis absorption spectra of the mixed solutions were tested on a uv spectrophotometer to study precursor compound III-2 vs HS^-The response activity of (2). The final concentration of precursor compound III-2 was 10.0. mu. mol/L, and the final concentration of NaHS was gradually increased from 0.0. mu. mol/L to 10.0. mu. mol/L.

FIG. 2 (A) shows the UV-visible absorption spectra of precursor compound III-2 after different concentrations of NaHS were added to the solution; (B) is a plot of the relative values of the absorbance at 355nm versus 580nm for III-2 as the NaHS concentration increases; the inset is a photograph of III-2 taken in the bright field before and after addition of 1 equivalent of NaHS to the solution.

As can be seen from FIG. 2, with HS^-The characteristic absorption peak of the precursor compound III-2 gradually decreased with increasing concentration, and a new absorption peak was gradually formed around 355 nm. When 1 equivalent of HS is added^-Then, the absorption peak of precursor compound III-2 at around 580nm almost completely disappeared. Furthermore, with HS^-The color of the solution gradually becomes lighter from the original blue color and finally becomes colorless, and the change of the solution color also indicates the precursor compounds III-2 and HS^-The addition process of (1). The results show that the precursor compound III-2 can react with HS^-And (4) high-efficiency addition.

Example 5

Water response H₂S release:

the probes I-2 or II-2 of examples 1 and 2 were taken, and 30. mu.L of the mother liquor diluted with DMSO to 1.0mmol/L (1mmol/L means that the concentration of each of the probes I-2 or II-2 was 1mmol/L) was added to 2970. mu.L of DMSO or ultrapure water, respectively, and mixed well, and after incubating at room temperature for 0.5 hour, the mixture was incubated with UVThe mixed solutions were tested by a spectrophotometer for uv-vis absorption spectra to study their dissociation and release of H₂Activity of S. The final concentrations of I-2 or II-2 were 10.0. mu. mol/L.

With reference to the above method, corresponding volumes of the products containing I-2 or II-2 of example 1 or 2 (probes I-2 or II-2 stored in the mixed solvent of example 1 or 2) were taken and added to ultrapure water so that their final concentrations were 25. mu. mol/L, and H was used₂S detection kit for detecting H released by S detection kit₂The efficiency of S.

FIG. 3 (A) shows an ultraviolet-visible light absorption spectrum of a precursor compound III-2 and a probe I-2 or II-2 in DMSO or water; (B) is a bright field photograph of III-2(1, 4), I-2(2, 5) or II-2(3, 6) in DMSO or water;

FIG. 4 (A) is a standard curve of absorbance versus hydrogen sulfide concentration; (B) is the concentration of hydrogen sulfide released by probe I-2 or II-2 triggered by water.

As can be seen from FIG. 3, addition of I-2 or II-2 to DMSO revealed that the absorption peak was still around 355nm, while almost no significant absorption peak appeared at around 580nm, corresponding to the characteristic absorption of III-2 itself, which is consistent with the results of example 3, indicating that I-2 or II-2 can be stably present in DMSO. Subsequently, after addition of I-2 or II-2 to water, it was found that the absorption peak around 355nm had almost completely disappeared, and accordingly a new absorption peak appeared at 580nm, and the absorption spectrum almost coincided with that of III-2 in water, indicating that water triggered dissociation of I-2 or II-2 into III-2 and restored the characteristic absorption peak of III-2. Further, it can be seen from FIG. 3 (B) that I-2 or II-2 was colorless in DMSO, and the color of the solution became blue after adding them to water. Thus, a change in solution color is also indicative of dissociation of I-2 or II-2.

As can be seen from FIG. 4, under the water trigger, 25. mu. mol/L of I-2 or II-2 released 16.4. mu. mol/L and 16.0. mu. mol/L of H, respectively₂S, indicating that the I-2 or II-2 as an intelligent probe can efficiently release the physiologically active molecule H through water response₂S。

Example 6

Na₂Cell proliferation assay for S and III-2:

fibroblast cells L929 were seeded into 96-well plates, 1 ten thousand cells were seeded per well, cultured in an incubator for 12 hours, and then Na-containing solutions having different concentrations were added to the respective wells₂S or III-2 medium 100 u L and continued to cultivate for 24 hours. After 24 hours, the medium was aspirated and washed with PBS to remove extracellular Na₂S or III-2, then 100. mu.L of CCK-8 working solution was added to each well and further incubated in an incubator for 2 hours. The absorbance at 450nm was then measured with a multifunctional microplate reader, and the cell viability was calculated from the ratio of the absorbances.

FIG. 5 shows different concentrations of Na₂Cell viability of L929 cells 24 hours after S or precursor compound III-2 treatment.

As can be seen from FIG. 5, Na₂S has proliferation promoting effect on fibroblast L929 in concentration range of 0-200 μmol/L, and is associated with Na₂The proliferation promoting effect tends to increase with an increase in the S concentration. III-2 has no obvious influence on the survival rate of L929 in the concentration range of 0-100 mu mol/L, which indicates that III-2 has good biocompatibility.

Example 7

Hydrogen sulfide donor I-2 rat wound repair promotion experiment:

the alginate dressing membrane was used to adsorb a corresponding volume of DMSO using a control of 0.5mmol/L and 1mmol/L hydrogen sulfide donor I-2 adsorbed under sterile conditions.

SD rats of 6 weeks old weighing 150-: blank group, wound surface is not treated; a control group, the wound surface is treated by a seaweed salt dressing film absorbing DMSO; treating the wound surface with 0.5mmol/L seaweed salt dressing film adsorbing 0.5mmol/L I-2; 1mmol/L group, the wound surface was treated with seaweed salt dressing film adsorbing 1mmol/L I-2. After wound surfaces with the diameter of 10mm are generated on the left side and the right side of the back of a rat, the materials are randomly combined and applied to the wound surfaces, and the wound surfaces are bound by the application. Wound healing was observed and recorded at 0, 3, 7 and 14 days after surgery, and wound area was calculated using Image J software. Rats were sacrificed by dislocation of cervical vertebrae 3 days, 7 days and 14 days after the operation, and the wound surface and the surrounding normal tissues were removed and fixed in 4% paraformaldehyde, followed by paraffin section and hematoxylin-eosin (H & E) staining to evaluate the effect of hydrogen sulfide donor I-2 on promoting rat wound repair.

FIG. 6 is a diagram showing the healing process of the wound surface in the blank group, the control group and the probe I-2 group (0.5mmol/L group and 1mmol/L group);

FIG. 7 is a bar graph of the rate of healing of wounds at various time points for the blank, control, probe I-2 groups (0.5mmol/L and 1mmol/L groups);

FIG. 8 is a representative H & E staining of the wound surface of the blank group, the control group, and the probe I-2 group (0.5mmol/L group and 1mmol/L group) at each time point, (B) is an enlarged view of the corresponding region in (A), and the yellow line is the boundary between the normal tissue and the wound surface.

As can be seen from FIG. 6, the 0.5mmol/L and 1mmol/L groups triggered H by the absorption of wound tissue exudate by the alginate dressing membrane on day 3 post-surgery₂And S is released and III-2 is generated, so that the seaweed salt dressing membrane is changed into purple black, the release of hydrogen sulfide is indicated, and self-monitoring is realized. 7 days after the operation, the wound surfaces of the 0.5mmol/L group and the 1mmol/L group are smaller than those of the blank group and the control group, and obvious difference is shown.

As can be seen from FIG. 7, the healing rates in the 0.5mmol/L group and the 1mmol/L group were 35.9%. + -. 1.5% and 34.9%. + -. 1.5%, respectively, 3 days after the operation, which were greater than those in the blank group and the control group. After 7 days, the healing rates of the 0.5mmol/L group and the 1mmol/L group reach 69.7% + -6.4% and 76.1% + -5.2%, which are greater than 59.5% + -2.0% and 59.1% + -4.4% of the blank group and the control group; wherein the healing rate of the 1mmol/L group is obviously greater than that of the blank group and the control group.

As can be seen from fig. 8, at day 3 post-surgery, the wounds of each group were covered with fibrin clots. Wherein, the inflammatory cells of the 0.5mmol/L group and the 1mmol/L group are relatively less, and a large amount of inflammatory cells exist at the wound surface of the blank group and the control group, which indicates that the wound surface is in serious inflammatory reaction. In addition, as shown by the red arrows in fig. 8(B), more new blood vessels were observed in both the 0.5mmol/L and 1mmol/L groups, and the wound was transporting a large amount of nutrients through the blood vessels to accelerate healing thereof; while little new blood vessels were observed in the blank group and the control group. This shows that in the early stage of wound repair, the intelligent probe I-2 can effectively inhibit inflammatory reaction and promote the generation of new vessels. On day 7 post-surgery, an ordered arrangement of a large number of spindle-shaped fibroblasts and a large number of extracellular matrix (ECM) was observed in the 0.5mmol/L and 1mmol/L groups, as well as vessels that germinated from the bottom of the wound and grew up as loops, and granulation tissue replaced the initial fibrin clot, indicating that the wound entered the proliferative phase. In addition, the number of blood vessels in the 1mmol/L group was reduced compared to the 0.5mmol/L group and there was a small amount of collagen deposition (as indicated by the green arrows in FIG. 8 (B)), indicating that the wound was progressing further from the proliferative phase into the remodeling phase. In contrast, the wounds of the blank and control groups still had inflammatory cells present, in which a small number of new blood vessels were formed, indicating that the wounds healed slowly. Fibroblast cells which are regularly arranged are observed in the control group, which shows that the strong hygroscopicity of the seaweed salt dressing membrane is beneficial to keeping the moist protective environment of the wound surface, and the healing of the wound surface is accelerated to a certain extent. On day 14 post-surgery, new epidermal tissue had formed in each group, with the epidermis being more intact in both the 0.5mmol/L and 1mmol/L groups. As shown by the blue arrows in FIG. 8(B), new glandular and follicular tissue had formed in the wound surface of 0.5mmol/L and 1mmol/L groups, and there was a large amount of collagen deposition, especially in the 1mmol/L group. The wound surface of the 0.5mmol/L group and the 1mmol/L group has almost no obvious difference from the surrounding normal tissues. Whereas only mature granulation tissue was observed in the blank group and the control group, no significant collagen deposition was observed, and the boundary between the wound surface and normal tissue in the two groups was clearly visible (as indicated by the yellow line in fig. 8 (a)).

The higher the content of the organic solvent is, the more stable the probe is; sodium sulfide can react with the precursor compound when the water content is below 40%, but the addition efficiency is higher when the water content is lower; the organic solvent content is more than 90% (the rest is water) and can be stored at low temperature. The probe of the present invention is stored and used at low temperature in the form of a mixed solution.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.