CN115607741B - Composite biological material for glaucoma surgery and application thereof - Google Patents
Composite biological material for glaucoma surgery and application thereof Download PDFInfo
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- CN115607741B CN115607741B CN202211100393.4A CN202211100393A CN115607741B CN 115607741 B CN115607741 B CN 115607741B CN 202211100393 A CN202211100393 A CN 202211100393A CN 115607741 B CN115607741 B CN 115607741B
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Classifications
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- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L2300/626—Liposomes, micelles, vesicles
Abstract
The invention relates to a composite biological material for glaucoma surgery and application thereof, and relates to the technical field of biomedical materials. A composite biological material for glaucoma surgery comprises a compact layer substrate and a loose layer; the porous layer comprises liposome, and is formed by mixing gel with high crosslinking degree and gel with low crosslinking degree, and is cast on a compact layer substrate, and the porous layer is dried to obtain the composite biological material, wherein the thickness ratio of the porous layer to the compact layer is 1:1-5:1. wherein, the compact layer plays a role in physical support and scar formation blocking; the loose layer is liposome containing antimetabolite, which can slowly release antimetabolite, and the loose layer absorbs water in the initial stage of operation to enlarge the filtering space, reduce the aggregation of postoperative inflammatory cells and blood coagulation factors, prevent the formation of scar at the operation site, gradually reduce the filtering space along with the degradation of gel with low crosslinking degree, and coordinate the functions of the two functions, so that the wound healing filtering channel is stably formed, and the operation failure rate is reduced.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a composite biological material for glaucoma surgery and application thereof.
Background
Glaucoma is a group of ocular diseases with characteristic optic nerve damage and visual field defects, and can also be defined as optic neuropathy with structural damage to the characteristic optic nerve and visual field defects. The normal intraocular pressure range of Chinese is 10-21 mmHg (mm Hg), most people can press optic nerves and blood vessels for supplying optic nerve nutrition when the intraocular pressure is increased beyond a threshold range, such as blood vessel automatic regulation function is reduced, when the intraocular pressure is increased beyond a blood vessel bearing range, the blood vessels cannot be regulated, and the normal metabolic requirement of the optic nerve cannot be met, so that the function of the optic nerve is reduced, the optic nerve is atrophic, the optic disc is in large and deep depression, and visual field visual glaucoma is typically changed. Pathological increases in ocular pressure are the first risk factor, and the intensity of ocular pressure and the resistance to ocular fundus optic nerve compression are closely related to the occurrence and progression of glaucomatous optic nerve atrophy and its characteristic visual field defects. Glaucoma is one of the most serious blinding eye diseases at present, and is irreversible.
At present, clinical treatment mainly depends on ocular pressure reduction, and conservative treatment such as the use of antihypertensive drugs and laser treatment have greatly advanced, but filterable surgery is still the first treatment method for certain glaucoma, and the purpose of the surgery is to manufacture a permanent fistula which does not heal, so that functional blebs can be formed, thereby reducing intraocular pressure and avoiding damage to optic nerves due to ocular hypertension. However, due to the inherent wound healing mechanism of the human body, more than 20% of patients often undergo surgery failure due to scarring of the filtration channels after filtration.
Scarring of the glaucoma surgical area is generally divided into 2 processes: (1) As the blood-ocular barrier is broken by surgery, plasma fibrin exudes, fibrin coagulates into blocks, deposits in the surgical field, blocks the filtration passages, and causes obstruction of aqueous humor filtration, leading to early postoperative failure. (2) The operation area has coagulum containing fibrin, fibronectin and blood platelet, which provides a support for the migration of inflammatory cells and fibroblasts, the fibroblasts proliferate and secrete collagen, the scar of the operation area heals, the function of the drainage aqueous humor of the filtration passage is reduced until the drainage aqueous humor is lost, and the later period of operation is failed.
For this reason, CN201510455055.6 discloses a composite biological matrix for glaucoma surgery and a method for preparing the same. The composite biological matrix is a double-layer composite structure formed by aging and freeze drying a compact layer substrate and a loose layer coated on the compact layer substrate, wherein the compact layer substrate is a cross-linked decellularized amniotic membrane or animal tissue membrane, the loose layer is a porous membrane prepared from a biological macromolecular material, and the loose layer takes the compact layer substrate as a support and can be regarded as a 'water storage sponge' of the composite biological matrix, and the porous membrane plays a role in dynamically regulating the outflow of aqueous humor mainly through an adsorption-extrusion function. However, the method cannot solve the problems that plasma fibrin exudes, fibrin coagulates into blocks, deposits in an operation area, blocks a filtration passage and causes obstruction of aqueous humor filtration.
Thus, there is an urgent need to provide a composite biomaterial for glaucoma surgery to prevent scarring and reduce the rate of surgical failure by forming a stable filtered channel.
Disclosure of Invention
In order to solve the technical problems, the invention provides a composite biological material for glaucoma surgery, which comprises a loose layer and a compact layer; the compact layer can play a role in physical support and scar formation blocking; the loose layer is liposome gel containing antimetabolite, which can slowly release antimetabolite to prevent scar formation. Meanwhile, the gel with low crosslinking degree and the gel with high crosslinking degree are compounded to form a loose layer, and due to the high water absorption of the gel with low crosslinking degree, the water absorption of the loose layer can increase the filtering space at the initial stage of operation, reduce the aggregation of inflammatory cells and coagulation factors after operation, prevent the formation of scar at the operation position, and gradually reduce the filtering space along with the degradation of the gel with low crosslinking degree; the two functions are coordinated, so that a filtering channel is formed stably, and the operation failure rate is reduced.
Meanwhile, the invention also provides a composite biological material for glaucoma surgery and application thereof in the postoperative treatment of glaucoma. The physical support and scar formation blocking effects of the compact layer are combined with the scar formation blocking effects of the gel with low crosslinking degree and the long-time antimetabolite release effects of the gel with high crosslinking degree, so that the medical treatment frequency and pain of patients can be reduced.
The invention provides a composite biological material for glaucoma surgery, which consists of a compact layer substrate and a loose layer; the porous layer comprises liposome, and is formed by mixing gel with high crosslinking degree and gel with low crosslinking degree, and is cast on a compact layer substrate, and the porous layer is dried to obtain the composite biological material, wherein the thickness ratio of the porous layer to the compact layer is 1:1-5:1.
preferably, the compact layer substrate is obtained by reducing and cleaning animal tissue membranes after being crosslinked for a plurality of times by different crosslinking agents.
Preferably, the gel with low crosslinking degree is obtained by crosslinking a mixed solution D of a biomacromolecule solution and a liposome containing an antimetabolite by a solution B of a crosslinking agent, and then washing, homogenizing and dialyzing;
the concentration of the biological macromolecule solution is 10-100mg/ml; in the mixed solution D, the volume-mass ratio of the biomacromolecule solution to the liposome containing the antimetabolite is 15:1-30:1;
the mass concentration of the cross-linking agent B solution is 0.5-2%, and the volume ratio of the cross-linking agent B solution to the mixed solution D is 1:5-1:15.
Preferably, the gel with high crosslinking degree is obtained by crosslinking a mixed solution D with a crosslinking agent C solution, and then washing, homogenizing and dialyzing the mixed solution D, wherein the mass concentration of the crosslinking agent C solution is 0.05-0.5%, and the volume ratio of the crosslinking agent C solution to the mixed solution D is 1:5-1:15.
Preferably, the loose layer is obtained by mixing gel with low crosslinking degree and gel with high crosslinking degree according to the mass ratio of 3:1-6:1.
Preferably, the animal tissue membrane is one of peritoneum, pericardium, small intestine submucosa and bladder matrix membrane of pig, cattle, horse and sheep sources.
Preferably, the multiple crosslinking is secondary crosslinking; the secondary crosslinking is as follows: placing the animal tissue membrane in a cross-linking agent A1 solution with the mass concentration of 0.5-5% for first cross-linking, cleaning after soaking treatment, and soaking in a cross-linking agent A2 solution with the mass concentration of 0.5-5% for second cross-linking; the cross-linking agent A1 is at least one selected from EDC, glutaraldehyde, epoxide and genipin; the cross-linking agent A2 is at least one selected from EDC, glutaraldehyde, epoxide and genipin.
Preferably, in the secondary crosslinking process, the mass ratio of the animal tissue membrane to the solution of the crosslinking agent A1 is 1:5-1:20, and the mass ratio of the animal tissue membrane to the solution of the crosslinking agent A2 is 1:5-1:20.
The invention also provides a preparation method of the composite biological material for glaucoma surgery, which comprises the following steps:
step one, preparing a compact layer substrate: immersing animal tissue membranes in a cross-linking agent A1 solution with the mass concentration of 0.5-5% for the first time, and cleaning after the immersing treatment; soaking in 0.5-5% cross-linking agent A2 solution for the second time, and soaking;
placing the animal tissue membrane subjected to secondary crosslinking into a buffer solution containing a reducing agent for three times of reduction, and then cleaning to obtain a compact layer substrate;
step two, preparing a loose layer: adding liposome containing antimetabolite with volume-mass ratio of 15:1-30:1 into 10-100mg/ml biomacromolecule solution to obtain mixed solution D; the biological macromolecules are one or more of recombinant collagen, hyaluronic acid and chitosan;
adding a cross-linking agent B solution with the mass concentration of 0.5-2% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree; the volume ratio of the cross-linking agent B solution to the mixed solution D is 1:5-1:15;
adding a cross-linking agent C solution with the mass concentration of 0.05-0.5% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree; the volume ratio of the solution of the cross-linking agent C to the mixed solution D is 1:5-1:15;
step three, preparation of a composite biological material: uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 3:1-6:1, casting on the compact layer substrate prepared in the step one, and drying to obtain the composite biological material.
The invention also provides application of the composite biological material for glaucoma surgery in the postoperative treatment of glaucoma.
The beneficial effects are that:
(1) In the composite biological material for glaucoma surgery, a compact layer is subjected to secondary crosslinking by adopting two crosslinking agents, wherein EDC crosslinking mainly connects carboxyl and amino on collagen of animal tissue membrane materials to form an amide bond, glutaraldehyde crosslinking mainly forms Schiff base by aldehyde groups and primary amino groups, epoxide crosslinking mainly reacts with hydroxyl groups and carboxyl groups to form ether bonds, and genipin crosslinking mainly forms heterocyclic amine by using olefinic carbon atoms and amino groups; through the two different crosslinking mechanisms, the animal tissue membrane material forms a more stable crosslinking structure, the degradation time of the animal tissue membrane in the body is prolonged, the degradation period is far longer than the period of post-operation fistula formation, and the scar formation of a post-operation filtration channel can be effectively inhibited.
(2) The low-crosslinking-degree gel and the high-crosslinking-degree gel are compounded to form a loose layer, the low-crosslinking-degree gel has high water absorbability, the water absorbability of the loose layer can increase the filtering space at the initial stage of operation, the aggregation of inflammatory cells and blood coagulation factors after operation is reduced, the formation of scars at the operation position is prevented, along with the degradation of the low-crosslinking-degree gel, the filtering space is gradually reduced, and the low-crosslinking-degree gel and the functions are coordinated, so that a wound healing filtering channel is stably formed, and the operation failure rate is reduced.
(3) The composite biological material for glaucoma surgery has two layers, wherein one layer is a compact layer and the other layer is a loose layer containing liposome; the loose layer is liposome gel containing antimetabolite, which can slowly release antimetabolite, wherein, the liposome combines gel matrixes with different crosslinking degrees to realize the double slow release function of antimetabolite, the gel degradation time with low crosslinking degree is short, the antimetabolite can be released in the tissue repair proliferation period, the scar formation is prevented, the gel degradation time with high crosslinking degree is long, the gel can still play a role after the tissue repair proliferation period is finished, the long-time release of antimetabolite is realized, and the scar formation of tissue is restrained in a long period. Avoiding frequent injection of antimetabolite after operation.
(4) The invention provides application of a composite biological material for glaucoma surgery in the postoperative treatment of glaucoma; the physical support and scar formation blocking effects of the compact layer are combined with the scar formation blocking effects of the gel with low crosslinking degree and the long-time antimetabolite release effects of the gel with high crosslinking degree, so that postoperative pain of glaucoma patients can be reduced, and meanwhile, the medical treatment frequency of the patients can be reduced through long-time antimetabolite release.
Drawings
In order to more clearly illustrate the technical solutions of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a cross-sectional scanning electron microscope of the product prepared in example 1;
FIG. 2 is a comparative graph showing the results of in vitro drug release tests of example 1 and comparative examples 2 to 3;
FIG. 3 is a comparative view of a 4w histological section taken after surgery of example 1 and comparative examples 2-3;
FIG. 4 is a 12w histological section of the product prepared in example 1 after surgery.
Detailed Description
In order to more fully understand the technical content of the present invention, the technical solution of the present invention will be further described and illustrated with reference to specific examples, but the embodiments of the present invention are not limited thereto.
In the invention, "%" refers to mass percent unless otherwise noted.
The composite biological material for glaucoma surgery consists of a compact layer and a loose layer, wherein the compact layer is an animal tissue membrane (pericardium, peritoneum, small intestine mucosa, conjunctiva, sclera and the like) and is crosslinked by a crosslinking agent, and the crosslinking agent is one or more of EDC, glutaraldehyde and epoxide. The porous layer comprises liposome, and is formed by mixing gel with high crosslinking degree and gel with low crosslinking degree, and is cast on a compact layer substrate, and is dried to obtain the composite biological material, wherein the thickness ratio of the porous layer to the compact layer is 1:1-5:1.
The loose layer is combined with an antimetabolite slow release carrier to realize long-term medicine slow release, reduce the discomfort of multi-frequency medicine injection and prevent scar formation of filtration blebs so as to achieve the aim of treating glaucoma.
The preparation method of the composite biological material for glaucoma surgery comprises the following steps:
step one, preparing a compact layer substrate: immersing animal tissue membrane in 0.5-5% cross-linking agent A1 solution for first cross-linking, immersing at 2-20deg.C for 4-12h, and cleaning; soaking in 0.5-5% cross-linking agent A2 solution for secondary cross-linking at 4-20deg.C for 4-24 hr, wherein the mass ratio of animal tissue membrane to cross-linking agent A1 solution is 1:5-1:20, and the mass ratio of animal tissue membrane to cross-linking agent A2 solution is 1:5-1:20; the cross-linking agent A1 is at least one of EDC, glutaraldehyde, epoxide and genipin, the cross-linking agent A2 is also at least one of EDC, glutaraldehyde, epoxide and genipin, and the cross-linking agent A1 and the cross-linking agent A2 are different;
placing the animal tissue membrane subjected to secondary crosslinking in a buffer solution containing a reducing agent for three times of reduction, and then cleaning (PBS solution, preferably pH 7 can be used in cleaning) to obtain a compact layer substrate; the reducing agent can be at least one of stannous chloride, oxalic acid, potassium borohydride, sodium borohydride and ethanol; the buffer solution may be selected from PBS solutions;
step two, preparing a loose layer: adding liposome containing antimetabolite with volume-mass ratio of 15:1-30:1 into 10-100mg/ml biomacromolecule solution to obtain mixed solution D; the biological macromolecules are one or more of recombinant collagen, hyaluronic acid and chitosan; wherein the antimetabolite is selected from mitomycin, 5-fluorouracil, colchicine, methotrexate, doxorubicin, daunorubicin, homoharringtonine, and paclitaxel;
adding a cross-linking agent B solution with the mass concentration of 0.5-2% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree; the volume ratio of the cross-linking agent B solution to the mixed solution D is 1:5-1:15; adding a cross-linking agent C aldehyde solution with the mass concentration of 0.05-0.5% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree; the volume ratio of the solution of the cross-linking agent C to the mixed solution D is 1:5-1:15; the crosslinking agent B and the crosslinking agent C are commonly used crosslinking agents, and can be specifically selected from EDC, glutaraldehyde, genipin, epoxide and the like; namely, the cross-linking agent B can be at least one selected from EDC, glutaraldehyde, genipin, epoxide and the like, and the cross-linking agent C can be at least one selected from EDC, glutaraldehyde, genipin, epoxide and the like;
it should be noted that the low and high judgment basis in the low and high crosslinking degree gels in the present invention is the crosslinking degree thereof, rather than the amount of the crosslinking agent added, and the amount of the crosslinking agent added is different depending on the kind of the crosslinking agent; for example, EDC crosslinking ability is relatively weak, and when added in a large amount, it may be a gel with a low degree of crosslinking; glutaraldehyde has relatively strong crosslinking capability, and can be added in relatively small amount to form gel with high crosslinking degree;
step three, preparation of a composite biological material: uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 3:1-6:1, casting on the compact layer substrate prepared in the step one, and drying to obtain the composite biological material.
Example 1
Step one, preparing a compact layer substrate: immersing the bovine pericardium subjected to cell and antigen removal in 3% EDC solution for primary crosslinking, immersing at 16 ℃ for 8 hours, and cleaning; soaking in 3% glutaraldehyde solution for secondary crosslinking, and soaking at 16deg.C for 16 hr, wherein the mass ratio of the bovine pericardium to EDC solution or glutaraldehyde solution is 1:10;
and (3) placing the bovine pericardium after secondary crosslinking in PBS (phosphate buffer solution) containing a reducing agent for three times of reduction, and then cleaning to obtain the compact layer substrate.
Step two, preparing a loose layer: adding mitomycin-containing liposome with the volume-mass ratio of 20:1 into 20mg/ml sodium hyaluronate solution to obtain a biological macromolecule-liposome mixed solution;
adding EDC solution with the concentration of 1.2% into the mixed solution, and performing crosslinking reaction by the volume ratio of the EDC solution to the mixed solution being 1:10 to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree;
adding glutaraldehyde solution with the concentration of 0.4% into the mixed solution, and performing a crosslinking reaction by using a volume ratio of the crosslinking agent to the mixed solution of 1:10 to form gel; reducing, cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree;
the loose layer comprises a low-crosslinking-degree gel and a high-crosslinking-degree gel;
step three, preparation of a composite biological material: and (3) uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 4:1, casting on the compact layer substrate prepared in the step (A), and drying to obtain the composite biological material.
FIG. 1 is a photograph of a cross-section of a product prepared in example 1, showing the structure of loose layers and dense layers.
Example 2
Step one, preparing a compact layer substrate: the pig peritoneum with cells and antigens removed is soaked in 1% genipin solution for the first time to crosslink, and is washed after soaking treatment for 12 hours at 4 ℃; soaking in 0.5% EDC solution for secondary crosslinking, wherein the mass ratio of animal tissue membrane to genipin solution or EDC solution is 1:18;
and placing the pig peritoneum subjected to secondary crosslinking into a PBS solution containing a reducing agent for three times for reduction, and then cleaning to obtain the compact layer substrate.
Step two, preparing a loose layer: adding a liposome containing 5-fluorouracil with the volume-mass ratio of 30:1 into 10mg/ml chitosan solution to obtain a biological macromolecule-liposome mixed solution;
adding an epoxide solution with the concentration of 0.8 percent into the mixed solution, and carrying out a crosslinking reaction by the epoxide solution and the mixed solution in a volume ratio of 1:15 to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree;
adding genipin solution with the concentration of 0.2% into the mixed solution, and performing a crosslinking reaction by the volume ratio of the crosslinking agent to the mixed solution of 1:15 to form gel; reducing, cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree;
the loose layer comprises a low-crosslinking-degree gel and a high-crosslinking-degree gel;
step three, preparation of a composite biological material: and (3) uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 3:1, casting on the compact layer substrate prepared in the step (A), and drying to obtain the composite biological material.
Example 3
Step one, preparing a compact layer substrate: immersing the cell-free antigen-removed submucosa of the small intestine of the pig in glutaraldehyde solution with the concentration of 4.5% for the first time, and cleaning after immersing at 20 ℃ for 6 hours; soaking in 4% epoxide solution for secondary crosslinking, wherein the mass ratio of the submucosa of the small intestine of the pig to glutaraldehyde solution or epoxide solution is 1:5;
and (3) placing the animal tissue membrane subjected to secondary crosslinking in PBS (phosphate buffer solution) containing a reducing agent for three times for reduction, and then cleaning to obtain the compact layer substrate.
Step two, preparing a loose layer: adding liposome containing antimetabolite with volume-mass ratio of 15:1 into 80mg/ml recombinant collagen solution to obtain biomacromolecule-liposome mixed solution; the biological macromolecules are one or more of recombinant collagen, hyaluronic acid and chitosan;
adding EDC solution with the concentration of 2% into the mixed solution, and carrying out crosslinking reaction by the volume ratio of the EDC solution to the mixed solution being 1:5 to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree;
adding genipin solution with the concentration of 0.5% into the mixed solution, and performing a crosslinking reaction by the volume ratio of the crosslinking agent to the mixed solution of 1:5 to form gel; reducing, cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree;
the loose layer comprises a low-crosslinking-degree gel and a high-crosslinking-degree gel;
step three, preparation of a composite biological material: and (3) uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 6:1, casting on the compact layer substrate prepared in the step (A), and drying to obtain the composite biological material.
Comparative example 1
EDC was used for both crosslinking of bovine pericardium and the rest of the procedure was the same as in example 1.
Comparative example 2
The low-crosslinking degree gel and the high-crosslinking degree gel are uniformly mixed according to the mass ratio of 1:1, and the rest steps are the same as in example 1.
Comparative example 3
The low-crosslinking degree gel and the high-crosslinking degree gel are uniformly mixed according to the mass ratio of 8:1, and the rest steps are the same as in example 1.
Comparative example 4
EDC solution with concentration of 0.1% was added to the mixed solution to prepare a gel with low crosslinking degree, and the rest was the same as in example 1.
Comparative example 5
EDC solution with concentration of 2.5% was added to the mixed solution to prepare a gel with low crosslinking degree, and the rest was the same as in example 1.
Comparative example 6
Glutaraldehyde solution with a concentration of 0.01% was added to the mixed solution to prepare a gel with a high degree of crosslinking, and the rest of the procedure was the same as in example 1.
Comparative example 7
Glutaraldehyde solution with a concentration of 1.0% was added to the mixed solution to prepare a gel with a high degree of crosslinking, and the rest of the procedure was the same as in example 1.
In vitro degradation tests were performed on bovine pericardial materials of example 1 and comparative example 1, and the results obtained are shown in table 1 below.
Table 1 in vitro degradation test of bovine pericardial material of example 1 and comparative example 1
| Group of | Degradation time h |
| Example 1 | 48.32±0.34 |
| Comparative example 1 | 27.99±0.40 |
Table 1 shows the results of in vitro degradation tests of bovine pericardium materials obtained in example 1 and comparative example 1 with 100U/ml type I collagenase, respectively: the in vitro enzymolysis time of the composite biological material of the example 1 after EDC and glutaraldehyde secondary crosslinking is far higher than that of the composite biological material of the comparative example 1 crosslinked by only using EDC, which shows that the product degradation time is greatly prolonged after two different crosslinking agents are secondarily crosslinked, and the continuous and long-acting physical support and scar formation blocking effects can be provided for the operation part.
The prepared examples 1 to 3 and comparative examples 1 to 7 were applied to the postoperative recovery stage after glaucoma surgery, and their effects on postoperative recovery were judged. Normal eye status without glaucoma served as a blank.
Table 2 below shows the post-operative 4W and 12W ocular pressure averages of application examples 1-3 and comparative examples 1-7 and the 4W and 12W ocular pressure averages of the blank.
TABLE 2 mean post-operative intraocular pressure
Table 2 shows results of tonometric measurements of 4w and 12w after the operation of the products of application examples 1 to 3 and comparative examples 1 to 7 and the blank. Wherein, fig. 4 shows that the product prepared in example 1 is obtained by histological sections of 12w materials after operation, the materials are complete and clear under a microscope, and meanwhile, no obvious inflammation and foreign body reaction are seen, which indicates that the product has good histocompatibility.
As shown in the results of table 2, the intraocular pressures of 4w and 12w are at normal level after the operation of the example, which is similar to the intraocular pressure of the blank control, and the application of the product of the example forms a stable filtering channel at the operation site, so that the normal physiological activities of eyes can be maintained; in comparative example 1, the post-operation 4w intraocular pressure is at a normal level, the post-operation 12w intraocular pressure is increased, and the in vitro degradation test of bovine pericardium material is combined, so that the compact layer of the product is degraded too quickly to provide continuous and long-acting physical support and scar formation blocking effects for an operation part.
In comparative examples 2-3, the intraocular pressure of 4w and 12w is higher after the operation in comparative example 2, the gel proportion with high crosslinking degree in the loose layer in the sample is too high, and the combination of the in vitro medicine release research result and the tissue inflammation condition of 4w after the operation shows that inflammatory cells can be aggregated in the tissue repair proliferation period, and meanwhile, under the condition that sufficient antimetabolite is not sustained, early scars can be formed at the operation part, so that the operation fails; comparative example 3 the post-operative 4w ocular pressure is at normal level, the post-operative 12w ocular pressure is increased, the low cross-linking degree gel proportion ratio in the loose layer of the sample is too high, which can lead to the too fast degradation of the material in vivo, and simultaneously, the anti-metabolic drug is released in large quantity in early stage in vivo, the problem of insufficient medium and long term dosage occurs, and simultaneously, the collapse of the filtration passage can be caused along with the degradation of the low cross-linking degree gel, which causes the operation failure.
In comparative examples 4-5, the intraocular pressure of 4w and 12w is higher after the operation of comparative example 4, the crosslinking degree of the gel with low crosslinking degree in the sample is too low, and the in vitro medicine release research result shows that the gel with low crosslinking degree is quickly degraded after being implanted into the body, so that the medicine is quickly released in early stage, the medicine release amount is reduced along with the completion of the degradation of the gel with low crosslinking degree, the medicine use amount is insufficient, and meanwhile, the space collapse of a filtration passage is caused along with the quick degradation of the gel with low crosslinking degree, and the recurrence of glaucoma is caused; comparative example 5 has high post-operation 4w and 12w intraocular pressure, and in the sample, the crosslinking degree of the gel with low crosslinking degree is too high, and the achieved treatment efficiency is similar to that of comparative example 2, namely, the release amount of the antimetabolite is insufficient in early stage after operation, and meanwhile, the water absorption of the loose layer is low, so that the formed filtration space is insufficient, and the operation fails.
In comparative examples 6 to 7, the post-operation 4w intraocular pressure of comparative example 6 was at a normal level, the post-operation 12w intraocular pressure was increased, the high crosslinking degree of the gel in this sample was too low, and the achieved therapeutic efficiency was similar to comparative example 3, i.e., problems of large release amount of the drug in the early stage after operation, insufficient medium-long term use amount, and collapse of the filtration passage were generated; comparative example 7 the post-operative 4w ocular pressure is at normal level, the post-operative 12w ocular pressure is increased, the gel crosslinking degree of high crosslinking degree is too high, so that the material degradation time is too long, the drug release amount is insufficient in the middle and long periods, and the operation fails.
The in vitro release test of the drugs was carried out on the prepared examples 1 to 3 and comparative examples 2 to 7, and graphs of the test results obtained are shown in FIG. 2. The sample of the example 1 can continuously and stably release medicines in the whole period, and the aggregation condition of inflammatory cells is observed by combining the intraocular pressure after operation, so that the use of the sample of the example can inhibit the formation of scar after operation.
In comparative example 2, the high-crosslinking gel proportion in the loose layer is higher, the drug release speed is slower, and the aggregation condition of inflammatory cells is observed in combination with 4w after operation, which indicates that the drug release cannot be matched with the tissue repair proliferation period, and meanwhile, the aggregation of inflammatory cells can cause scar formation, so that the operation fails; in comparative example 3, the low-crosslinking gel in the porous layer had a high proportion, and the gel degraded too rapidly, causing premature massive release of the drug, but insufficient support to inhibit long-term scarring, while the low-crosslinking gel degraded too rapidly, resulting in collapse of the filtration channels, and thus also causing recurrence of glaucoma.
In comparative example 4, the low-crosslinking degree gel in the loose layer is too low in crosslinking degree, rapid degradation of the low-crosslinking degree gel in the early stage after operation is accompanied by rapid release of the drug, and then the release amount is reduced, so that the dosage is insufficient, which indicates that the release of the drug cannot be matched with the tissue repair proliferation period; in comparative example 5, the gel crosslinking degree of the low crosslinking degree in the bulk layer was too high, and the effect was similar to that of comparative example 2.
In comparative example 6, the gel crosslinking degree of the high crosslinking degree in the bulk layer was too low, and the effect was similar to that of comparative example 3; in comparative example 7, the gel with high crosslinking degree in the loose layer has too high crosslinking degree, and early gel degradation and drug release after operation can meet the requirement of drugs, and the gel degradation is slower in medium and long term, and the drug release amount is insufficient, so that the failure of operation can be caused.
FIG. 3 shows histological sections of 4w material after surgery, wherein A is the sample of example 1, B is the sample of comparative example 2, and C is the sample of comparative example 3. In the example 1 and the comparative example 3, the postoperative loose layer has high water absorption, increases the filtering space, and further does not cause inflammatory cell aggregation; in comparative example 2, however, the porous layer had a low water absorption due to a high gel ratio, and the resulting filtration space was small, resulting in inflammatory cell aggregation.
The foregoing description is only illustrative of the present invention and is not intended to limit the scope of the invention, and all equivalent structures or equivalent processes or direct or indirect application in other related technical fields are included in the scope of the present invention.
Claims (7)
1. A composite biological material for glaucoma surgery is characterized by comprising a compact layer substrate and a loose layer; the porous layer comprises liposome, and is formed by mixing gel with high crosslinking degree and gel with low crosslinking degree, and is cast on a compact layer substrate, and the porous layer is dried to obtain the composite biological material, wherein the thickness ratio of the porous layer to the compact layer is 1:1-5:1, a step of;
the gel with low crosslinking degree is obtained by crosslinking a mixed solution D of a biological macromolecule solution and a liposome containing an antimetabolite by a solution B of a crosslinking agent, and then cleaning, homogenizing and dialyzing;
the concentration of the biological macromolecule solution is 10-100mg/ml; in the mixed solution D, the volume-mass ratio of the biomacromolecule solution to the liposome containing the antimetabolite is 15:1-30:1;
the mass concentration of the cross-linking agent B solution is 0.5-2%, and the volume ratio of the cross-linking agent B solution to the mixed solution D is 1:5-1:15;
the gel with high crosslinking degree is obtained by crosslinking a mixed solution D by a crosslinking agent C solution, and then washing, homogenizing and dialyzing the mixed solution D, wherein the mass concentration of the crosslinking agent C solution is 0.05-0.5%, and the volume ratio of the crosslinking agent C solution to the mixed solution D is 1:5-1:15;
the loose layer is obtained by mixing gel with low crosslinking degree and gel with high crosslinking degree according to the mass ratio of 3:1-6:1.
2. The composite biomaterial for glaucoma surgery according to claim 1, wherein the dense layer substrate is obtained by reducing and cleaning animal tissue membranes after multiple crosslinking with different crosslinking agents.
3. The composite biomaterial for glaucoma surgery according to claim 2, wherein the antimetabolite is one selected from mitomycin, 5-fluorouracil, colchicine, methotrexate, doxorubicin, daunorubicin, homoharringtonine, paclitaxel.
4. The composite biomaterial for glaucoma surgery according to claim 2, wherein the animal tissue membrane is one of peritoneum, pericardium, small intestine submucosa, bladder matrix membrane of porcine, bovine, equine, ovine origin.
5. The glaucoma surgical composite biomaterial of claim 2, wherein the multiple crosslinks are secondary crosslinks; the secondary crosslinking is as follows: placing the animal tissue membrane in a cross-linking agent A1 solution with the mass concentration of 0.5-5% for first cross-linking, cleaning after soaking treatment, and soaking in a cross-linking agent A2 solution with the mass concentration of 0.5-5% for second cross-linking; the cross-linking agent A1 is at least one selected from EDC, glutaraldehyde, epoxide and genipin; the cross-linking agent A2 is at least one selected from EDC, glutaraldehyde, epoxide and genipin.
6. The composite biomaterial for glaucoma surgery according to claim 5, wherein in the secondary crosslinking process, the mass ratio of the animal tissue membrane to the solution of the crosslinking agent A1 is 1:5-1:20, and the mass ratio of the animal tissue membrane to the solution of the crosslinking agent A2 is 1:5-1:20.
7. The method for preparing a composite biomaterial for glaucoma surgery as defined in claim 6, comprising the steps of:
step one, preparing a compact layer substrate: immersing animal tissue membranes in a cross-linking agent A1 solution with the mass concentration of 0.5-5% for the first time, and cleaning after the immersing treatment; soaking in 0.5-5% cross-linking agent A2 solution for the second time, and soaking;
placing the animal tissue membrane subjected to secondary crosslinking into a buffer solution containing a reducing agent for three times of reduction, and then cleaning to obtain a compact layer substrate;
step two, preparing a loose layer: adding liposome containing antimetabolite with volume-mass ratio of 15:1-30:1 into 10-100mg/ml biomacromolecule solution to obtain mixed solution D; the biological macromolecules are one or more of recombinant collagen, hyaluronic acid and chitosan;
adding a cross-linking agent B solution with the mass concentration of 0.5-2% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with low crosslinking degree; the volume ratio of the cross-linking agent B solution to the mixed solution D is 1:5-1:15;
adding a cross-linking agent C solution with the mass concentration of 0.05-0.5% into the mixed solution D for cross-linking reaction to form gel; then cleaning, homogenizing and dialyzing to obtain gel with high crosslinking degree; the volume ratio of the solution of the cross-linking agent C to the mixed solution D is 1:5-1:15;
step three, preparation of a composite biological material: uniformly mixing the gel with low crosslinking degree and the gel with high crosslinking degree according to the mass ratio of 3:1-6:1, casting on the compact layer substrate prepared in the step one, and drying to obtain the composite biological material.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104958791A (en) * | 2015-07-29 | 2015-10-07 | 陕西博与再生医学有限公司 | Composite biological matrix for glaucoma surgery and preparation method thereof |
| WO2019031776A1 (en) * | 2017-08-08 | 2019-02-14 | 오스템임플란트 주식회사 | Composite collagen membrane in which collagen layers having different biodegradation rates are layered, and production method therefor |
| CN112553785A (en) * | 2020-11-16 | 2021-03-26 | 山东奥精生物科技有限公司 | Double-layer guided tissue regeneration membrane and preparation method thereof |
Family Cites Families (2)
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| KR20180117211A (en) * | 2010-01-22 | 2018-10-26 | 알러간, 인코포레이티드 | Intracameral sustained release therapeutic agent implants |
| US20160038632A1 (en) * | 2014-08-07 | 2016-02-11 | Massachusetts Instutite Of Technology | Adaptive Drug Delivery from an Artificial Polymer Skin with Tunable Properties for Tissue Engineering |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104958791A (en) * | 2015-07-29 | 2015-10-07 | 陕西博与再生医学有限公司 | Composite biological matrix for glaucoma surgery and preparation method thereof |
| WO2019031776A1 (en) * | 2017-08-08 | 2019-02-14 | 오스템임플란트 주식회사 | Composite collagen membrane in which collagen layers having different biodegradation rates are layered, and production method therefor |
| CN112553785A (en) * | 2020-11-16 | 2021-03-26 | 山东奥精生物科技有限公司 | Double-layer guided tissue regeneration membrane and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| A tissue-engineered human trabecular meshwork hydrogel for advanced glaucoma disease modeling;Haiyan Li 等;Experimental Eye Research;第205卷;1-18 * |
| 治疗青光眼的新型给药系统研究进展;张惠 等;中国医药工业杂志;第51卷(第11期);1364-1370 * |
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