CN109045063B

CN109045063B - In-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism and preparation method and application thereof

Info

Publication number: CN109045063B
Application number: CN201811211077.8A
Authority: CN
Inventors: 薛伟明; 魏荣; 李可欣; 许宁侠; 温惠云; 黄赛朋; 杨华; 潘士印
Original assignee: Northwestern University
Current assignee: Shaanxi Liangshuangtong Medical Technology Development Co ltd
Priority date: 2018-10-17
Filing date: 2018-10-17
Publication date: 2021-01-26
Anticipated expiration: 2038-10-17
Also published as: CN109045063A

Abstract

The invention discloses an in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism and a preparation method and application thereof, belonging to the technical field of medical products. The composite hydrogel is liquid sol with fluidity at room temperature, and the sol is placed into the lacrimal passage of a human body through punctum injection. By utilizing the gel phase transition characteristic of the sol which has sensitive response to the body temperature, the in-situ gelation and embolism of the sol in the lacrimal passage are realized, a small amount of tears secreted by the affected eye are retained in the orbit, and the purpose of keeping the ocular surface moist for a long time is achieved. The body temperature responsiveness of the water-soluble chitosan and sodium glycerophosphate compound is utilized, and alginate and calcium chloride are added into the compound to adjust the mechanical property, the sol-gel phase transition time, the mechanical strength and the like of the compound, so that the compound is simple to operate, mild in reaction condition, low in cost, good in biocompatibility, small in wound, capable of filling lacrimal passages with any shape and size, and good in relieving effect on xerophthalmia patients.

Description

In-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medical products, and relates to an in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism and a preparation method and application thereof.

Background

Dry eye disease (dry eye disease) is an eye disease with symptoms such as decreased stability of tear film and abnormal quality and quantity of tear fluid due to various causes such as congenital lacrimal gland hypoplasia, trachoma and ocular corneal sclerosis. The essence of this disease is a reduction in the number and density of goblet cells in the tear film, causing changes in the quality or quantity of tear secretion. The clinical symptoms mainly comprise dry eyes, itching and the like, and the long-term accumulation of the symptoms can cause deep pathological changes of the keratoconjunctiva of a patient and cause vision loss in severe cases. Epidemiological investigation results show that the prevalence rate of the dry eye disease is related to regions, occupation, age and gender, for example, the prevalence rate of Asian race is high, and the prevalence rate of the dry eye disease is high when a video terminal is used as the occupation, such as a teacher, staff and a driver. It has been found that dry eye patients develop a vicious cycle, i.e., decreased lacrimal secretion → decreased ocular surface sensitivity → decreased response of lacrimal gland to ocular surface irritation → decreased lacrimal secretion. The clinical treatment method of dry eye disease includes artificial tear substitution, operation transplantation, lacrimal passage embolism implantation, etc. In the research of artificial tears, the rainbow of scenic spots and the like discover that carbomer gel has longer residence time on the ocular surface and can effectively relieve dry eye symptoms; matsuo et al found that trehalose ophthalmic drug can stabilize the phospholipid bilayer structure of keratoconjunctival epithelial cells, increase the density of conjunctival goblet cells, and enhance the ability of keratoconjunctival epithelial cells to withstand a dry environment. Nevertheless, the main problems of artificial tears in clinical applications are: the physiological amount of tears required for the ocular surface cannot be stably maintained, the antimicrobial agent and preservative contained in the preparation aggravate ocular surface damage, and frequent use can reduce the treatment compliance of patients, etc. In 1998 Murube et al first proposed the treatment of dry eye by transplantation of the labial minor salivary glands, and then Geer-ling et al also carried out similar studies. In follow-up visit to a plurality of patients with xerophthalmia treated by the labial gland transplantation, the surgically transplanted glands still survive, the lacrimal secretion of the patients is increased, and the disordered ocular surface structure is improved, but the success rate of the surgery is extremely low.

At present, the treatment scheme with the most obvious curative effect is lacrimal duct embolization, and after the lacrimal punctum is embolized by adopting a micro-size lacrimal duct embolus, natural tears can be increased to provide longer moistening for the ocular surface; at the same time, increased natural tearThe liquid can stimulate the secretion of tears and promote the survival of goblet cells, thereby increasing the stability of the tear film; the medicine loss can be reduced and the medicine taking times can be reduced by embolizing the lacrimal passage. When the lacrimation or other complications occur in the treatment of the lacrimal duct embolism, the embolus can be taken out at any time, so that the irreversibility of the traditional operation is avoided. The study of lacrimal duct plugs began in the early 20 th century, and Foulds in 1961 blocked the canaliculus with nondegradable endophytic plants, and in 1975, Freeman Plug, designed by Freeman, became the prototype for the preparation of future lacrimal duct plugs, and the current foreign brand of lacrimal duct plugs: SmartPlug^TM、Form Fit^TM、Snug Plugs^TM、Soft Plug^TM、Tears Naturale^TM

Extend^TM、

And the like. At present, lacrimal passage embolism products used for clinically treating xerophthalmia in China all depend on imports, are high in price, and greatly increase the economic burden of patients. In addition, the imported lacrimal passage suppository product designed based on European and American people cannot completely adapt to the eye anatomical characteristics of Asian people, and the problem of operation failure caused by displacement or falling-off of the embolus in clinical application often exists.

Therefore, the development of lacrimal passage suppository products with independent intellectual property rights in China, good lacrimal passage embolism effect and moderate cost has important significance and development value.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide an in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism, and a preparation method and application thereof, wherein the composite hydrogel has good fluidity at room temperature, is gelatinized by responding to the environmental temperature, has controllable shape and size, better biocompatibility and antibacterial activity and low toxic and side effects; the preparation method has the advantages of cheap and easily obtained raw materials, mild reaction conditions, simple and convenient operation and stable embolism performance; the composite hydrogel can be applied as a medicine for treating xerophthalmia.

In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:

the invention discloses an in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism, which comprises the following raw materials in percentage by mass:

1 to 15 percent of water-soluble chitosan, 4 to 10 percent of sodium glycerophosphate, 1 to 5 percent of alginate and 0.03 to 0.1 percent of calcium chloride.

Preferably, the molecular weight of the water-soluble chitosan is 100000-200000, and the substitution degree is 70% -95%.

Preferably, the water-soluble chitosan is carboxymethyl chitosan, chitosan quaternary ammonium salt or chitosan hydrochloride.

Preferably, the sodium glycerophosphate is beta-sodium glycerophosphate, alpha-sodium glycerophosphate or alpha beta-sodium glycerophosphate.

Preferably, the alginate is sodium alginate or calcium alginate.

Preferably, the calcium chloride is anhydrous calcium chloride or calcium chloride dihydrate.

The invention also discloses a preparation method of the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism, which comprises the following steps:

1) weighing the raw materials according to a metering ratio, and dissolving water-soluble chitosan in deionized water to prepare a water-soluble chitosan aqueous solution;

2) dissolving sodium glycerophosphate in deionized water to prepare a sodium glycerophosphate aqueous solution;

3) mixing the water-soluble chitosan aqueous solution prepared in the step 1) and the sodium glycerophosphate aqueous solution prepared in the step 2) according to the weight ratio of 1: (0.6-1.2) to obtain a mixed solution A, adding alginate into the mixed solution A, and stirring to dissolve the alginate sufficiently to obtain a mixed solution B;

4) dissolving calcium chloride in deionized water to prepare a calcium chloride aqueous solution, adding the prepared calcium chloride aqueous solution into the mixed solution B prepared in the step 3), and fully and uniformly stirring to obtain a mixed solution C;

5) and transferring the mixed solution C into a mold, and carrying out gelation treatment at 35-39 ℃ to prepare the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism.

Preferably, in the step 1), the water-soluble chitosan accounts for 4-15% of the water-soluble chitosan aqueous solution by mass percent; in the step 2), the sodium glycerophosphate accounts for 8-20% of the sodium glycerophosphate aqueous solution by mass; in the step 4), the calcium chloride accounts for 0.5-5% of the calcium chloride aqueous solution by mass percent.

The invention also discloses application of the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism in preparation of a medicine for treating xerophthalmia.

Preferably, the medicine is a medicine for relieving dry eye by blocking lacrimal canaliculus with the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel.

Preferably, the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism is a liquid sol with fluidity at room temperature, has gel phase transition characteristics with sensitive response to physiological temperature, and can generate gelation phase transition in lacrimal passage to form in-situ gelation embolism.

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

the invention discloses an in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism, which is a liquid sol with fluidity at room temperature, the composite hydrogel material utilizes the sensitive response characteristic of a water-soluble chitosan and sodium glycerophosphate compound at physiological temperature, calcium chloride and alginate are added into the system to change the sol-gel phase transformation rate, the mechanical strength and the like, the flowable sol is placed into the lacrimal passage through lacrimal punctum injection in vitro, the sol system is subjected to gelation transformation through the response to the temperature change, the method has the advantages of lower operation cost, smaller wound to a patient, capability of filling lacrimal passage cavities with any shape and size with the sol, better antibacterial performance and low toxic and side effects, and has good relieving and treating effects on the xerophthalmia patients.

The preparation method of the composite hydrogel disclosed by the invention is low in cost, simple to operate and mild in reaction conditions, and can fill lacrimal cavities with any shapes and sizes as required, so that the aim of relieving and treating the symptoms of xerophthalmia patients is fulfilled.

Furthermore, the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism is a liquid sol with fluidity at room temperature, has gel phase transition characteristics with sensitive response to physiological temperature, and can generate gelation phase transition in the lacrimal passage to form in-situ gelation embolism.

Drawings

FIG. 1 is a flow chart of an experiment for sample preparation according to the present invention;

FIG. 2 is a diagram of gelation mechanism of chitosan quaternary ammonium salt (HTCC) -GP;

FIG. 3 shows Na-Alg-Ca²⁺A gelation mechanism diagram of (1);

FIG. 4 shows HTCC-GP and Na-Alg-Ca²⁺A gelation mechanism diagram of (1);

FIG. 5 is a photograph showing the morphology of a polymer before gelation;

FIG. 6 is a photograph showing the morphology of the polymer after gelation;

FIG. 7 is a graph showing the effect of HTCC concentration on gelation time;

FIG. 8 is a microstructure (100X) view of a gel section;

FIG. 9 is a graph showing the results of inhibition of E.coli and S.aureus activity by the gel;

FIG. 10 is a graph showing the effect of hydrogel lacrimal plugs on the height of the lacrimal river under rabbit eyes;

FIG. 11 is a diagram of a right eye conjunctival blot cell examination of a rabbit without hydrogel embolization;

FIG. 12 is a photograph of a right eye conjunctival blot of rabbits implanted with hydrogel plugs.

Detailed Description

In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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.

It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

The invention is described in further detail below with reference to the accompanying drawings:

example 1

Referring to fig. 1, the experimental process of the present invention includes the following steps: accurately weighing 6 wt% of HTCC powder, dissolving the HTCC powder in deionized water to prepare HTCC) aqueous solution, preparing 14 wt% of beta-sodium Glycerophosphate (GP) aqueous solution in the same way, and mixing the HTCC powder and the deionized water in a volume ratio of 1: 0.7 of HTCC aqueous solution and GP aqueous solution are mixed and stirred uniformly, and then 2 percent Na-Alg powder is added into the HTCC-GP mixed aqueous solution and is fully stirred and dissolved. To the HTCC-GP-Na-Alg mixed aqueous solution of example 1 was added 3% CaCl₂Stirring the water solution uniformly, transferring into a mold, and placing the mold in a constant-temperature water bath kettle at 37 ℃ to ensure that the water solution is gelatinized and converted into hydrogel.

The mechanism in the gelation process is shown in fig. 2, fig. 3 and fig. 4, fig. 2 is a gelation mechanism diagram of HTCC-GP, and water molecules form a water film on HTCC long chains at room temperature through the combination of hydrogen bonds; the addition of GP molecules enables the system to have temperature sensitivity, phosphate groups with negative charges in the GP molecules have electrostatic interaction with quaternary amino groups with positive charges on HTCC molecules, and are combined with partial protonated amino groups on the HTCC molecules through electrostatic interaction, so that more GP molecules are attached to the HTCC long chain. In GP molecules, due to the induction effect of electrons, C atoms adjacent to phosphate groups carry positive charges, O atoms connected with C atoms carry negative charges, and H atoms connected with O atoms carry positive charges. The O atom with negative charge between GP molecules attached on the HTCC chain forms a more stable hydrogen bond with the H atom with positive charge of another molecule GP.

FIG. 3 shows Na-Alg-Ca²⁺Gelation mechanism diagram, 1 Ca²⁺Forms a complex with 2 guluronic acid (G unit) G segments in the Na-Alg molecular chain segment through 4 coordination bonds, namely, an egg lattice structure (figure 3). FIG. 4 shows HTCC-GP system with Na-Alg-Ca²⁺The system forms an interpenetrating network structure through random interpenetration.

Example 2

HTCC-GP-Na-Alg-Ca of example 1²⁺The mixed aqueous solution is transferred into a penicillin bottle, and the result is shown in fig. 5, the mixed solution is a mixed solution with fluidity at room temperature, and the in-situ injectable temperature-sensitive response chitosan quaternary ammonium salt composite hydrogel for lacrimal passage embolism, which loses fluidity, is obtained after gelation at 37 ℃ (fig. 6).

Example 3

Respectively dissolving 1 wt%, 3 wt%, 5 wt%, 7 wt% and 9 wt% of HTCC powder in deionized water to prepare an HTCC aqueous solution, and similarly preparing a 15% GP aqueous solution according to a volume ratio of 1: 0.8 preparing HTCC-GP mixed aqueous solution, transferring the mixed aqueous solution into a mould, gelating at 37 ℃ to obtain the in-situ injectable temperature-sensitive response chitosan quaternary ammonium salt hydrogel for lacrimal passage embolism, and observing the gelating time by the phenomenon that a test tube inversion method system in the embodiment 1 loses fluidity, wherein the specific implementation method comprises the following steps: placing a parallel penicillin bottle filled with the same formula in a constant temperature of 37 ℃, inclining the penicillin bottle at intervals of 10s by using a stopwatch, observing the flowing phenomenon of the system, and recording the time as the phase transformation time of the gel when the system loses fluidity or the liquid level does not change after inclining. The effect of the change in the HTCC concentration of the component on the gelation time was observed (fig. 7), and it is seen from fig. 7 that the gelation time decreased first and then increased with the increase in the HTCC concentration, indicating that the optimum value of the HTCC concentration was determined from the gelation time of the system, and further indicating that the system could achieve the purpose of adjusting the gelation rate by adjusting the proportional concentration of the component.

Example 4

The sample prepared in example 3 is freeze-dried, the section of the dried gel sample is taken, the microscopic morphology of the HTCC gel sample is observed by a scanning electron microscope after gold spraying (fig. 8 scanning electron microscope picture (× 100)), fig. 8 shows that the HTCC hydrogel has a three-dimensional reticular porous structure, and the structure enables movement of more small molecules to be unimpeded, so that normal physiological metabolism of a human body is not affected.

Example 5

HTCC-GP-Na-Alg-Ca prepared in example 3²⁺The mixed aqueous solution was poured into a mold having a side length of 1cm, and then placed at a constant temperature of 37 ℃ for a while, and the gel was taken out. The gel sample was compressed by a universal material tester at a compression rate of 70% of the original size to give a compressive modulus of elasticity of 53kPa, 51.9kPa, 52.4kPa, respectively, and the compressive modulus values can be used to represent the mechanical strength of the gel sample, thereby indicating that the HTCC hydrogel has sufficient mechanical strength to maintain its stable presence in the lacrimal canaliculus when the lacrimal canaliculus of a human body vibrates or contracts.

Example 6

Adding 1g of Yeast extract, 0.5g of Tpyptone and 1g of NaCl powder into 100mL of distilled water, stirring and dissolving to prepare a liquid culture medium, then placing the liquid culture medium into a vertical pressure steam sterilizer for sterilization at 121 ℃ for 30min, and then placing the liquid culture medium on a clean bench for standby.

Respectively inoculating Escherichia coli and Staphylococcus aureus to 2 sterilized liquid culture media with sterile inoculating needle, culturing at 37 deg.C for 24h in constant temperature oscillator, and placing in-4 deg.C refrigerator for use.

The hydrogel prepared in example 5 was cut into gel sheets having the same shape and size, weighed to make the weight of each sheet approximately the same, and the sample was sterilized by irradiation with an ultraviolet lamp for 30min (both sides were sterilized). Adding 20mL of sterilized culture solution and 0.1mL of bacterial solution into a 50mL sterile conical flask, then respectively adding gel slice samples and respectively marking, leaving a bottle without the samples and the bacterial solution as blank samples, then placing the bottle into a constant temperature oscillator at 37 ℃ for culturing for 24 hours, and respectively measuring the absorbance value of each sample on an ultraviolet spectrophotometer at 650 nm. The test results are shown in fig. 9, the HTCC hydrogel has inhibitory effect on the activity of both escherichia coli and staphylococcus aureus at different times, and the inhibitory effect of the HTCC hydrogel on bacteria tends to be stable with the time.

Example 7

Each of the aqueous raw material solutions prepared in example 3 was mixed by sterile filtration to obtain HTCC-GP-Na-Alg-Ca²⁺The aqueous solution was mixed and a certain amount of the mixed solution was sucked up with a syringe after sterilization.

Selecting 3 New Zealand white rabbits with no pathological changes or inflammations, such as cornea, conjunctiva and the like, adopting self contrast, and dropwise adding a fluorescein solution into a conjunctival sac to explore whether the lacrimal passage of the New Zealand white rabbits is smooth or not and remove the new Zealand white rabbits with the obstructed lacrimal passage. The HTCC mixed aqueous solution is placed into a right eye lacrimal passage through punctum injection, a left eye lacrimal passage is used as a contrast, the injected HTCC mixed aqueous solution fills the lacrimal passage firstly, and then gelation occurs to block the lacrimal passage in response to temperature change. After the rabbit eye is implanted for a period of time, the lower lacrimal river height of the rabbit eye is detected and analyzed through an anterior segment coherence tomography (AS-OCT), and the result is shown in fig. 10, it is seen from fig. 10 that the lower lacrimal river height of the rabbit eye is increased along with implantation of the hydrogel plug, and is higher than the lower lacrimal river height of a left eye which is compared with the lower lacrimal river height of a left eye which is self-controlled, and the lacrimal river height of an experimental group is 286 micrometers after 31 days and is still higher than the lacrimal river height before implantation, so that the accumulated amount of tears of the rabbit eye is increased after implantation of the HTCC hydrogel, and the effect of relieving and treating the xerophthalmia is achieved.

Example 8

Each of the aqueous raw material solutions prepared in example 3 was mixed by sterile filtration to obtain an HTCC-GP-Na-Alg-Ca2+ mixed aqueous solution, and a predetermined amount of the mixed solution was sucked up by a syringe after sterilization.

Selecting 3 New Zealand white rabbits with no pathological changes or inflammations, such as cornea, conjunctiva and the like, adopting self contrast, and dropwise adding a fluorescein solution into a conjunctival sac to explore whether the lacrimal passage of the New Zealand white rabbits is smooth or not and remove the new Zealand white rabbits with the obstructed lacrimal passage.

The HTCC mixed aqueous solution is placed into a right eye lacrimal passage through punctum injection, a left eye lacrimal passage is used as a contrast, the injected HTCC mixed aqueous solution fills the lacrimal passage firstly, and then gelation occurs to block the lacrimal passage in response to temperature change. Pathological changes in the mucus layer of the tear film of rabbit eyes were observed by conjunctival blot cytology (CIC) after a period of time. Firstly, opening the eyelids of rabbit eyes by using ***e, printing conjunctival epithelial cells by using cellulose acetate filter paper, and putting the conjunctival epithelial cells into 10% formalin; after PAS staining, dehydration and drying, the obtained product was observed under an optical microscope. The conjunctival blot cytogram (FIG. 11) of the right eye of the rabbit without hydrogel embolization was compared with the conjunctival blot cytogram (FIG. 12) of the right eye of the rabbit with hydrogel embolization, which showed more conjunctival goblet cells in the sampling region, a large number of neutrophils and eosinophils, a small number of inclusion bodies in the cytoplasm, and a small number of nuclei lines. Therefore, the hydrogel lacrimal passage suppository has a promoting effect on the proliferation of the conjunctiva goblet cells of the rabbit eyes and has no inflammatory reaction.

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims

1. The in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism is characterized by comprising the following raw materials in percentage by mass: 3-7% of chitosan quaternary ammonium salt, 4-10% of sodium glycerophosphate, 1-5% of alginate and 0.03-0.1% of calcium chloride; the molecular weight of the chitosan quaternary ammonium salt is 100000-200000, and the substitution degree is 70% -95%;

the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism has a three-dimensional reticular porous structure.

2. The in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism according to claim 1, wherein the sodium glycerophosphate is beta-sodium glycerophosphate, alpha-sodium glycerophosphate or alpha beta-sodium glycerophosphate.

3. The in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism according to claim 1, wherein the alginate is sodium alginate or calcium alginate.

4. The in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism according to claim 1, wherein the calcium chloride is anhydrous calcium chloride or calcium chloride dihydrate.

5. The preparation method of the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism according to any one of claims 1 to 4, which is characterized by comprising the following steps:

1) accurately weighing the raw materials according to a metering ratio, and dissolving chitosan quaternary ammonium salt in deionized water to prepare a chitosan quaternary ammonium salt solution;

3) mixing the chitosan quaternary ammonium salt solution prepared in the step 1) and the sodium glycerophosphate aqueous solution prepared in the step 2) according to the weight ratio of 1: (0.6-1.2) to obtain a mixed solution A, adding alginate into the mixed solution A, and stirring to dissolve the alginate sufficiently to obtain a mixed solution B;

5) and transferring the mixed solution C into a mold, and carrying out gelation treatment at 35-39 ℃ to prepare the in-situ injectable temperature-sensitive response hydroxypropyl chitosan composite hydrogel for lacrimal passage embolism.

6. The preparation method according to claim 5, wherein in the step 1), the mass percentage of the quaternary ammonium salt of chitosan in the quaternary ammonium salt solution of chitosan is 4% -15%; in the step 2), the mass percent of the sodium glycerophosphate in the sodium glycerophosphate aqueous solution is 8-20%; in the step 4), the calcium chloride accounts for 0.5-5% of the calcium chloride aqueous solution by mass.

7. The use of the in-situ injectable temperature-sensitive responsive water-soluble chitosan composite hydrogel for lacrimal passage embolism according to any one of claims 1 to 4 in the preparation of a medicament for treating dry eye.

8. The use of claim 7, wherein the medicament is a medicament for alleviating dry eye by blocking lacrimal canaliculus with the in situ injectable temperature-sensitive responsive water-soluble chitosan composite hydrogel;

the in-situ injectable temperature-sensitive response water-soluble chitosan composite hydrogel for lacrimal passage embolism is liquid sol at room temperature, has fluidity and gel phase change characteristic of sensitive response to body temperature, and can form in-situ gelation embolism in lacrimal passage.