CN114404676B - Self-healing hydrogel with heat insulation function and application thereof - Google Patents
Self-healing hydrogel with heat insulation function and application thereof Download PDFInfo
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Abstract
The invention relates to self-healing hydrogel with a heat insulation function and application thereof in preparation of a self-healing hydrogel isolating agent. The self-healing hydrogel is prepared from water-soluble chitosan, a cross-linking agent and a solution. The self-healing hydrogel release agent disclosed by the invention has excellent injectability, biocompatibility, biodegradability and conformability, can spontaneously heal into a whole after being extruded by an injection device, can conform to irregular tissue gaps, is not easy to disperse, has an excellent isolation protection effect in tumor thermal ablation, is simple to prepare and convenient to operate, and has huge application potential in the field of clinical thermal ablation.
Description
Technical Field
The invention belongs to the technical field of biological materials and hydrogel preparation, relates to self-healing hydrogel with a heat insulation function and application thereof, and particularly relates to self-healing hydrogel with a heat insulation function and application thereof in preparation of a tumor thermal ablation isolating agent.
Background
In recent years, ultrasound-guided tumor thermal ablation therapy is an important means for tumor therapy by virtue of the advantages of being minimally invasive, efficient and the like. The traditional separant is usually physiological saline, but when applied to tumor thermal ablation, the traditional separant has obvious disadvantages, including easy dispersion to surrounding tissue gaps, high heat transfer speed, undesirable heat insulation effect and the like. Therefore, a release agent having an excellent heat insulating effect has been a hot point of research.
Disclosure of Invention
The invention aims to provide a self-healing hydrogel with a heat insulation function, aiming at the defects of a physiological saline insulator in clinical tumor thermal ablation, wherein the self-healing hydrogel insulator is simple to prepare, has low solid content, and has good biocompatibility and biodegradability, injectability, self-healing property and conformability, in-situ retention property and heat insulation property.
The invention also aims to provide the application of the self-healing hydrogel with the heat insulation function in preparing the tumor thermal ablation isolating agent.
To this end, the invention provides, in a first aspect, a self-healing hydrogel having a thermal insulation function, which is obtained by reacting a water-soluble chitosan with a cross-linking agent in a solution.
According to the invention, the self-healing hydrogel has a heat insulation function and is injectable.
In some embodiments of the present invention, the water-soluble chitosan includes one or more of hydroxyethyl chitosan, carboxymethyl chitosan, carboxyethyl chitosan, hydroxypropyl chitosan; preferably, the chitosan has a deacetylation degree of 85.0 to 99.9% and a weight average molecular weight of 5000 to 100,000.
In other embodiments of the invention, the cross-linking agent is a PEG derivative with an aldehyde group at the end group, and has a weight average molecular weight of 1000 to 10,000.
In the invention, the solution comprises one or more of sterile water, physiological saline and phosphate buffer solution.
In a second aspect, the invention provides a method for preparing the self-healing hydrogel according to the first aspect, which comprises mixing a water-soluble chitosan solution and a cross-linking agent solution at room temperature to perform a chemical covalent cross-linking reaction to obtain the self-healing hydrogel; preferably, the volume ratio of the chitosan solution to the cross-linking agent solution is (1-3): 1.
In some embodiments of the invention, the concentration of the water-soluble chitosan solution is 10-30mg/mL.
In other embodiments of the present invention, the concentration of the crosslinking agent is 5-40mg/mL.
In still other embodiments of the present invention, the time for the crosslinking reaction is 10-20min.
In a third aspect, the present invention provides a thermal tumor ablation spacer formed from the self-healing hydrogel according to the first aspect of the present invention or the self-healing hydrogel prepared by the preparation method according to the second aspect of the present invention, wherein the thermal tumor ablation spacer has a thermal insulation function and is injectable, and is injected into a thermal tumor ablation site with a retention time of 5 to 7day; injecting the mixture into the tumor thermal ablation part at normal temperature, wherein the interface temperature between the tumor and the isolating agent in the operation is 37-45 ℃, and the interface temperature between the isolating agent and the isolated tissue in the operation is 36.5-40 ℃.
In a fourth aspect, the present invention provides a self-healing hydrogel according to the first aspect of the present invention or prepared by the preparation method according to the second aspect of the present invention, for use in preparing the thermal ablation barrier for tumor according to the third aspect of the present invention.
According to the invention, the application comprises the use of an injection device as a tumor thermal ablation spacer after extrusion of the self-healing hydrogel; preferably, the injection device comprises a syringe and an endoscope catheter.
In the invention, the tumor thermal ablation isolating agent can be spontaneously healed to synthesize complete hydrogel and is conformed to irregular tissue gaps for tumor thermal ablation isolation protection.
The invention has the beneficial effects that: the self-healing hydrogel separant is simple to prepare, has good biocompatibility, biodegradability and injectability, and can be extruded by an injector or an endoscope catheter to be spontaneously healed to synthesize complete hydrogel. The hydrogel release agent is endowed with excellent injectability and self-healing property by chemical covalent crosslinking, so that a larger shear gap is avoided, and the heat loss is effectively reduced, thereby having excellent heat insulation effect; the cross-linking agent has the characteristics of high latent heat, thermal stability and the like, and further enhances the heat insulation performance of the self-healing hydrogel release agent. In addition, the self-healing hydrogel release agent has conformability, can spontaneously change the shape to conform to irregular tissue gaps, is not easy to disperse, and has sufficient in-situ retention time; easy operation and convenient use.
Drawings
The invention is described in further detail below with reference to the attached drawing figures:
fig. 1 is an infrared heat transfer picture of the self-healing hydrogel release agent prepared by the invention and physiological saline.
FIG. 2 is a lower view of dark ultrasound with normal saline of the self-healing hydrogel release agent prepared by the present invention used for microwave ablation of thyroid nodules; wherein a and c are dark zone ultrasound views immediately upon injection; b and d are dark field ultrasound views under ablation.
Detailed Description
In order that the invention may be readily understood, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings. However, before the invention is described in detail, it is to be understood that this invention is not limited to particular embodiments described. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.
Unless otherwise defined, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.
As mentioned above, the physiological saline solution of the clinical existing tumor thermal ablation isolating agent has the defects of easy dispersion to the gaps of surrounding tissues, high heat transfer speed, undesirable heat insulation effect and the like. To solve these problems, the present inventors have conducted extensive studies on tumor thermal ablation isolation techniques.
The inventor researches and discovers that the hydrogel is a gel taking water as a dispersion medium, and a macromolecule with an hydrophilic/hydrophobic structure forms a 3D space network structure through physical crosslinking or chemical crosslinking, so that the hydrogel has good water absorbability, biocompatibility and biodegradability. The self-healing hydrogel has unique advantages compared with the traditional hydrogel as a novel injectable hydrogel, for example, the self-healing hydrogel can spontaneously heal to be in a complete shape after being broken by injection, is conformed to irregular tissue gaps, and can be applied to the fields of drug sustained release delivery and the like. However, none of the existing studies and techniques mention the application of self-healing hydrogels to thermal barriers.
The inventor further researches and discovers that the self-healing injectable hydrogel prepared by the reaction of the water-soluble chitosan and the cross-linking agent in the solution has the thermal conductivity close to that of water, but has the advantages of slow thermal diffusion, large thermal capacity, capability of absorbing a large amount of heat and excellent heat insulation function. Based on the above, the present inventors have studied and designed the use of such self-healing injectable hydrogel as a tumor thermal ablation insulating agent, thereby obtaining the present invention.
Therefore, the self-healing hydrogel having a heat insulating function according to the first aspect of the present invention is obtained by reacting a water-soluble chitosan with a crosslinking agent in a solution, and has a heat insulating function and injectability.
In the invention, the water-soluble chitosan comprises one or more of hydroxyethyl chitosan, carboxymethyl chitosan, carboxyethyl chitosan and hydroxypropyl chitosan; preferably, the chitosan has a deacetylation degree of 85.0 to 99.9% and a weight average molecular weight of 5000 to 100,000.
In the invention, the cross-linking agent is a PEG derivative with an aldehyde group at the end group, preferably DF-PEG 4000 (di-4-aldehyde polyethylene glycol benzoate) having a weight average molecular weight of 1000-10,000.
In the invention, the solution comprises one or more of sterile water, physiological saline and phosphate buffer solution, and is preferably physiological saline.
It should be understood by those skilled in the art that the above-mentioned "self-healing hydrogel having thermal insulation function, which is obtained by reacting water-soluble chitosan with a cross-linking agent in a solution" means that the water-soluble chitosan and the cross-linking agent are contacted in a solution (or under liquid phase conditions) to undergo a cross-linking reaction, and the two solutions can be directly added into the solution to undergo a reaction, or the above-mentioned solutions can be used as solvents to prepare a water-soluble chitosan solution and a cross-linking agent solution, which are then further mixed to undergo a cross-linking reaction.
In a second aspect of the present invention, there is provided a method for preparing the self-healing hydrogel according to the first aspect of the present invention, which comprises mixing a water-soluble chitosan solution with a cross-linking agent solution at room temperature to perform a chemical covalent cross-linking reaction to obtain the self-healing hydrogel.
In the invention, the water-soluble chitosan solution is formed by dissolving water-soluble chitosan in a solvent, and the concentration of the water-soluble chitosan solution is 10-30mg/mL, preferably 30mg/mL.
In the present invention, the aqueous crosslinking agent solution is formed by dissolving a crosslinking agent in a solvent, and the concentration of the crosslinking agent solution is 5 to 40mg/mL, preferably 5 to 10mg/mL, and more preferably 10mg/mL.
In some embodiments of the invention, the volume ratio of chitosan solution to the crosslinker solution is (1-3): 1, preferably 3:1.
In the invention, the solvent comprises one or more of water, normal saline and phosphate buffer solution, and the normal saline is preferred.
According to the invention, the time of the crosslinking reaction is 10-20min, preferably 15-20min, and more preferably 15min.
In some preferred embodiments of the present invention, the temperature of the crosslinking reaction is room temperature.
In a third aspect of the present invention, there is provided a thermal tumor ablation barrier agent formed from the self-healing hydrogel according to the first aspect of the present invention or the self-healing hydrogel prepared by the preparation method according to the second aspect of the present invention, the thermal tumor ablation barrier agent having a thermal insulation function and being injectable.
In some embodiments, the tumor thermal ablation spacer is injected into the tumor thermal ablation site, wherein the spacer has an area of 1-4cm 2 The horizontal and vertical spacing distance of the isolation region is 1-2cm, and the vertical spacing is 1-2cm; the retention time is 5-7day; injecting the mixture into the tumor thermal ablation part at normal temperature, wherein the interface temperature between the tumor and the isolating agent in the operation is 37-45 ℃, and the interface temperature between the isolating agent and the isolated tissue in the operation is 36.5-40 ℃.
In a fourth aspect, the present invention provides a self-healing hydrogel according to the first aspect of the present invention or prepared by the preparation method according to the second aspect of the present invention, for use in preparing a thermal ablation barrier for tumor according to the third aspect of the present invention. Therefore, the tumor thermal ablation insulating agent can also be called a self-healing hydrogel insulating agent with a heat insulation function or a tumor thermal ablation self-healing hydrogel insulating agent.
According to the present invention, the application includes extrusion of the self-healing hydrogel into a thermal ablation site using a syringe, endoscopic catheter, or the like, as a tumor thermal ablation spacer.
In the invention, the tumor thermal ablation isolating agent can be spontaneously healed to synthesize complete hydrogel and is conformed to irregular tissue gaps for tumor thermal ablation isolation protection.
Research results show that the self-healing hydrogel release agent has excellent injectability, biocompatibility, biodegradability and conformability, can spontaneously heal into a whole after being extruded by an injection device, can conform to irregular tissue gaps, is not easy to disperse, has an excellent isolation and protection effect in tumor thermal ablation, is simple to prepare and convenient to operate, and has huge application potential in the field of clinical thermal ablation.
III example
The present invention will be specifically described below with reference to specific examples. The experimental methods described below are, unless otherwise specified, all routine laboratory procedures. The experimental materials described below, unless otherwise specified, are commercially available.
Example 1:
preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting sterile water, taking 3 volumes of the water-soluble chitosan solution with the volume of 20mg/mL, adding 1 volume of the cross-linking agent solution with the volume of 10mg/mL, and mixing and reacting at room temperature for 20min to obtain the self-healing hydrogel isolating agent.
Example 2:
preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting sterile water, taking 3 volumes of the water-soluble chitosan solution with the volume of 20mg/mL, adding 1 volume of the cross-linking agent solution with the volume of 30mg/mL, and mixing and reacting at room temperature for 10min to obtain the self-healing hydrogel isolating agent.
Example 3:
preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting sterile water, taking 3 volumes of 30mg/mL water-soluble chitosan solution, adding 1 volume of 10mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 15min to obtain the self-healing hydrogel isolating agent.
Example 4:
preparing the water-soluble chitosan solution and the cross-linking agent solution by using physiological saline, taking 3 volumes of 20mg/mL water-soluble chitosan solution, adding 1 volume of 10mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 20min to obtain the self-healing hydrogel isolating agent.
Example 5:
preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting physiological saline, taking 3 volumes of 20mg/mL water-soluble chitosan solution, adding 1 volume of 30mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 10min to obtain the self-healing hydrogel isolating agent.
Example 6:
preparing the water-soluble chitosan solution and the cross-linking agent solution by using normal saline, taking 3 volumes of 30mg/mL water-soluble chitosan solution, adding 1 volume of 10mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 15min to obtain the self-healing hydrogel isolating agent.
Example 7:
and preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting a phosphate buffer solution, taking 3 volumes of the water-soluble chitosan solution with the volume of 20mg/mL, adding 1 volume of the cross-linking agent solution with the volume of 10mg/mL, and mixing and reacting at room temperature for 20min to obtain the self-healing hydrogel isolating agent.
Example 8:
and preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting a phosphate buffer solution, taking 3 volumes of 20mg/mL water-soluble chitosan solution, adding 1 volume of 30mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 10min to obtain the self-healing hydrogel isolating agent.
Example 9:
and preparing the water-soluble chitosan solution and the cross-linking agent solution by adopting a phosphate buffer solution, taking 3 volumes of 30mg/mL water-soluble chitosan solution, adding 1 volume of 10mg/mL cross-linking agent solution, and mixing and reacting at room temperature for 15min to obtain the self-healing hydrogel isolating agent.
Example 10:
comparative example: normal saline (conventional release agent).
Example 11
For the heat transfer experiment, the self-healing hydrogel release agent obtained in example 5 and the physiological saline solution of comparative example were placed in a cylindrical container, and a heating rod (100W, 15s) was inserted into the center of the container. The heat transfer process (heat transfer images of 1 st s, 7 th s, 15 th s) of the recording medium was photographed using a thermal infrared imager (FOTRIC 225 s), and the results are shown in fig. 1. As can be seen from fig. 1, the heat transfer area of the self-healing hydrogel release agent is significantly smaller than that of normal saline, indicating that the self-healing hydrogel release agent has excellent thermal insulation effect.
Example 12:
for the beagle thyroid nodule thermal ablation experiment at room temperature, the self-healing hydrogel release agent prepared in example 5 above and the physiological saline of the comparative example were used as release protectors for thyroid microwave ablation, and the lower view of the dark area ultrasound (color doppler ultrasound, SAMSUNG, korea) thereof is shown in fig. 2. As can be seen from fig. 2, a distinct isolation zone around the thyroid gland appeared after saline injection (fig. 2 a). At the time of ablation, saline had already diffused to the surrounding tissue (fig. 2 b). After injection of the self-healing hydrogel release agent, the formed isolation zone remained unchanged during ablation (fig. 2c and 2 d), indicating that the self-healing hydrogel release agent has excellent in-situ retentivity without repeated injection during ablation. Test results show that the retention time of the normal saline is 5min, and the retention time of the self-healing hydrogel isolating agent is 5-7day, which is obviously longer than that of the normal saline.
The area of the isolation region of the tumor thermal ablation part injected with the self-healing hydrogel is 1.29cm 2 The isolation region has a transverse spacing distance of 1.65cm, a longitudinal spacing distance of 1.16cm, and an intra-operative isolation region area of 1.24cm 2 The retention time is 5-7day, the gel release agent is injected into the tumor thermal ablation part at normal temperature, the interface temperature between the tumor and the gel release agent (the temperature of the gel release agent at the proximal end of the thyroid gland) is 41.8 ℃, and the gel release agent is separated from the tumorThe interfacial temperature between tissues (distal thyroid gel insulator temperature) was 37.4 ℃.
The area of the isolation region injected with physiological saline into the tumor thermal ablation part is 1.23cm 2 The isolation region has a transverse spacing distance of 1.79cm, a longitudinal spacing distance of 1.05cm, and an intra-operative isolation region area of 0.23cm 2 The retention time is 5min, the tissue is injected to a tumor thermal ablation operation part at normal temperature, the interface temperature between the tumor and the physiological saline isolating agent (the temperature of the physiological saline at the proximal end of the thyroid gland) is 50.5 ℃, and the interface temperature between the physiological saline isolating agent and the isolated tissue (the temperature of the physiological saline at the distal end of the thyroid gland) is 42.7 ℃.
Test results show that the self-healing hydrogel has excellent capabilities of isolating heat and protecting adjacent tissues.
Example 13:
for the endoscopic catheter injection experiment, the self-healing hydrogel release agent prepared in the above examples 1-9 can be extruded through the endoscopic catheter and can spontaneously heal to form complete hydrogel, and the heat insulation result of the tumor thermal ablation part injected through the endoscopic catheter is substantially the same as that obtained by using an injector.
The above examples show and describe the basic principles, preparation and use of the present invention. It will be understood by those skilled in the art that the present invention is not limited by the embodiments described above, which are intended to illustrate the principles of the invention and not to limit the scope of the invention in any way, and that various changes and modifications can be made without departing from the scope of the invention as defined by the appended claims.
Claims (7)
1. The application of self-healing hydrogel in preparing a tumor thermal ablation isolating agent;
the self-healing hydrogel is obtained by reacting water-soluble chitosan and a cross-linking agent in a solution, has a heat insulation function and is injectable;
the cross-linking agent is di-4-aldehyde polyethylene glycol benzoate, and the weight-average molecular weight of the cross-linking agent is 1000-10,000;
the tumor thermal ablation isolating agent has a heat insulation function and injectability, is injected to a tumor thermal ablation part, and has a retention time of 5-7day; injecting the mixture into the tumor thermal ablation part at normal temperature, wherein the interface temperature between the tumor and the isolating agent in the operation is 37-45 ℃, and the interface temperature between the isolating agent and the isolated tissue in the operation is 36.5-40 ℃.
2. The use of claim 1, wherein the water-soluble chitosan comprises one or more of hydroxyethyl chitosan, carboxymethyl chitosan, carboxyethyl chitosan, hydroxypropyl chitosan; the deacetylation degree of the chitosan is 85.0-99.9%, and the weight average molecular weight is 5000-100,000.
3. The use according to claim 1, wherein the solution comprises one or more of sterile water, physiological saline and phosphate buffered saline.
4. The use according to claim 1, wherein the self-healing hydrogel is prepared by mixing a water-soluble chitosan solution with a cross-linking agent solution at room temperature to perform a chemical covalent cross-linking reaction to prepare the self-healing hydrogel; the volume ratio of the chitosan solution to the cross-linking agent solution is (1-3): 1.
5. The use according to claim 4, wherein the concentration of the water-soluble chitosan solution is 10-30mg/mL; and/or, the concentration of the cross-linking agent is 5-40mg/mL; the time of the crosslinking reaction is 10-20min.
6. The use according to any one of claims 1-5, wherein the use comprises using an injection device to extrude the self-healing hydrogel for use as a tumor thermal ablation barrier; the injection device comprises an injector and an endoscope guide pipe.
7. The use of claim 6, wherein the tumor thermal ablation barrier is capable of self healing into a complete hydrogel and conforming to irregular tissue spaces for tumor thermal ablation barrier protection.
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