CN104130437A - Dural/spinal dural biological patch material, preparation method and application thereof - Google Patents
Dural/spinal dural biological patch material, preparation method and application thereof Download PDFInfo
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- CN104130437A CN104130437A CN201410373258.6A CN201410373258A CN104130437A CN 104130437 A CN104130437 A CN 104130437A CN 201410373258 A CN201410373258 A CN 201410373258A CN 104130437 A CN104130437 A CN 104130437A
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Abstract
The invention provides a dural/spinal dural biological patch material, a preparation method and application thereof. The obtained biological patch material is especially suitable for repair of dural/spinal dural defect, can maintain stable and non-degradable within 6 months-2 years after being implanted into the human body or animals, and can degrade gradually after more than 2 years. While ensuring certain mechanical properties and liquid seepage resistance, the biological patch material also has biocompatibility, has greatly reduced risks of postoperative complications and side effects, and meets the special requirements for dural/spinal dural defect repair materials.
Description
Technical field
The invention belongs to organizational engineering biomaterial for medical purpose technical field, be specifically related to a kind of hard brain/spinal meninges biology patching material and its preparation method and application.
Background technology
Endocranium and endorchis are respectively functional membrane tissues important between brain and skull, backbone and spinal cord, have the identical biological characteristics function of seeking peace.Endocranium attaches cranial cavity inner face, links and loosens with braincap, is combined closely with basis cranii; Endocranium stretches out at cranial nerve cranium place, divide a word with a hyphen at the end of a line and be combined with neural tunicle, foramen magnum periphery to downward be endorchis.Endocranium is consistent with spinal dural function and structure, and endocranium is for the protection of cerebral tissue, and endorchis is for the protection of myeloid tissue.
Wound, tumour, inflammation, neurosurgery or other brainpan spinal diseases etc. all can cause hard brain/spinal meninges damaged.The illnesss such as the damaged meeting of hard brain/spinal meninges causes that epilepsy, spinal fluid leak outside, intracranial infection, meninx bulging and disordered brain function.As in basis cranii surgical operation,, once there is damaged cerebrospinal rhinorrhea and the otorrhea of easily causing in basis cranii endocranium relative thin and being combined closely with basion.The tumour such as meningioma, chordoma of basis cranii usually corrodes and infiltrates to skull and endocranium around, and in the time that tumor resection is performed the operation, the tumour of excision also can be attached part meninx and skull conventionally, causes postoperative generation cerebrospinal leak and intracranial infection.
Damaged for treating hard brain/spinal meninges, in prior art normal adopt hard brain/spinal meninges equivalent material to implant damaged part to repair hard brain/spinal meninges damaged.Along with the high speed development of medical technology, all many-sides such as the dura defect that be more and more widely used in dural substitutes endocranium open decompression in the infiltration, surgical procedure of the operation of brain convex surface, wound coup injury, tumour and some geneogenous factors cause, as having 10%～15% to need hard brain/spinal meninges equivalent material to repair dura defect, also report that this ratio is up to 30% in brain surgery operation of opening cranium patient.Therefore,, in order to meet clinical needs, the suitable hard brain/spinal meninges repair materials that conducts a research is most important.
But often there is many deficiencies in existing hard brain/spinal meninges equivalent material, usually cause serious complication problem in the process using.The for example problem aspect degradation speed control after hard brain/spinal meninges equivalent material is implanting, if the excessive velocities of hard brain/spinal meninges equivalent material degraded, tear edge declines, cause anti-fluid seepage effect to decline, cause the illnesss such as hard brain/spinal meninges and surrounding tissue adhesion, infection, hemorrhage, cerebrospinal leak and epilepsy; If but hard brain/spinal meninges equivalent material do not degrade for a long time, cause again it not mate with tissue regeneration, biocompatibility is poor, easily causes what is more inflammatory reaction, connective tissue proliferation parcel, infection and the problem such as hemorrhage.Therefore, for hard brain/spinal meninges equivalent material, the control aspect the degradation speed in implant into body or animal body is very important.
But, in the technique with the hard brain/spinal meninges of natural membranes material produce equivalent material in the past, when mould material is after freeze-drying radiation, collagen protein in natural membranes material can significantly be destroyed, and mould material tear edge is declined, and in order better to slough cell and fat, the reagent of de-cell, antigen and the fat using is relatively large to the destruction of film, other mechanical properties of natural membranes material are significantly reduced, finally cause its degradation speed to be accelerated, anti-fluid seepage effect declines.In order to reduce the degradation speed of biological tissue's repair materials, in prior art, conventionally adopt linking agent to process mould material, improve its mechanical property, reduce its degradation speed, it can not degraded for a long time in human body or animal body.But for for repairing the equivalent material that hard brain/spinal meninges is damaged, because its special physiological location and function have determined in prior art for the treatment of the linking agent in the tissue renovation material at other positions or the damaged equivalent material of hard brain/spinal meninges and be not suitable for hard brain/spinal meninges patching material, the main problem existing comprises: after (1) linking agent is processed, repair materials is not degraded for a long time in human body or animal body, after causing it in implant into body or animal body, biocompatibility is poor, easily produces disadvantageous side effect; (2) stability of linking agent is low, most of linking agents all have than the group that is easier to reaction, this group causes linking agent unstable, and then cause repair materials in the short period of time, to be degraded in human body or animal body, cause the anti-fluid seepage effect of repair materials to decline, and then cause a series of post-operative complication, bring adverse influence; (3) linking agent toxicity is high, is difficult to control the finally residual quantity on product of linking agent, affects the healthy of implanted human or animal.Therefore, in prior art, need a kind of biology patching material that is more applicable to the requirement of hard brain/spinal meninges repairing degradation property of exploitation badly.
Summary of the invention
For this reason, technical problem to be solved by this invention is in prior art after hard brain/spinal meninges equivalent material implant into body that degradation speed is too fast or crosses to be all unfavorable for repairing slowly the problem that hard brain/spinal meninges is damaged, and then a kind of hard brain/spinal meninges biology patching material that is applicable to hard brain/spinal meninges defect repair degraded requirement is provided, and open its preparation method and application.
For solving the problems of the technologies described above, the invention provides a kind of preparation method of hard brain/spinal meninges biology patching material, comprise that getting animal membrane is organized as starting material, and be placed in turn the step that Racemic glycidol ethers epoxy crosslinking agent solution that surfactant soln, alkali lye, peroxide solutions, mass concentration are 1-5wt% and phosphate buffered saline buffer soak, and by after the described membrane tissue material lyophilize after above steps is soaked through irradiation sterilization, to obtain final product.
The preparation method of described hard brain/spinal meninges biology patching material, in the step of described Racemic glycidol ethers epoxy crosslinking agent solution soaking, described linking agent is propanetriol-diglycidyl-ether and/or diglycidyl ether of ethylene glycol.
The preparation method of described hard brain/spinal meninges biology patching material, in the step of described Racemic glycidol ethers epoxy crosslinking agent solution soaking, described soak time is 15h-10d.
Preferably, in the linking agent that described membrane tissue material is 2-4wt% in concentration, soak 20h.
Preferred, in the linking agent that described membrane tissue material is 2wt% in concentration, soak.
The preparation method of described hard brain/spinal meninges biology patching material, in described step of soaking containing surfactant soln, the mass concentration of described tensio-active agent is 0.1-5wt%, and controls described solution temperature and be 0-25 DEG C and soak.
The preparation method of described hard brain/spinal meninges biology patching material, in the step of described dipping by lye, the concentration of described alkali lye is 0.1-5mol/L, and controls described solution temperature and be 5-15 DEG C and soak.
Preferably, in the step of described dipping by lye, membrane tissue material is added in the sodium hydroxide solution that concentration is 1mol/L and soaks 2h, and to control solution temperature be 8 DEG C.
The preparation method of described hard brain/spinal meninges biology patching material, in the step that described peroxide solutions soaks, the mass concentration of described peroxide solutions is 1-5wt%.
Preferably, in the step of soaking at described peroxide solutions, it is immersion treatment 1-5h in 2wt% peroxide solutions that the membrane tissue material after immersion is added to concentration.
Preferably, described peroxide solutions is superoxol.
The preparation method of described hard brain/spinal meninges biology patching material, the step of described irradiation sterilization is electron beam irradiation sterilization, irradiation metering is 10-30KGy.
Preferably, the metering of the irradiation of described electron beam is 25KGy.
Preferably, in the step of soaking at described surfactant soln, described tensio-active agent is at least one in TritonX_100, Tween80 or Tween40.
Preferably, described animal membrane histogen material is 6 months following pigs of birth or the membrane tissue material of ox.
In the step that described phosphate buffered saline buffer soaks, the pH value of described phosphate buffered saline buffer is 5.0-8.0.
Preferably, in the step of soaking at described phosphate buffered saline buffer, the membrane tissue after described immersion adds pH to soak at least 5h in 6.8 phosphate buffered saline buffer.
The invention provides a kind of hard brain/spinal meninges biology patching material being prepared by above-mentioned method.
The present invention also provide a kind of described hard brain/spinal meninges biology patching material preparation prevent or reduce tissue adhesion's material, hard brain/spinal meninges damage of causing because of wound or tumour for material, treatment or the prevention of repairing impaired hard brain/spinal meninges tissue or damaged material, treatment or prevention be because of the application of the damaged cerebrospinal leak causing of brain/spinal meninges, infected material firmly.
The present invention also provides the purposes of a kind of Racemic glycidol ethers epoxy crosslinking agent for the preparation of hard brain/spinal meninges biology patching material.
Technical solutions according to the invention have following advantage compared to existing technology:
(1) hard brain/spinal meninges biology patching material preparation method of the present invention, utilize mass concentration for 1-5wt% Racemic glycidol ethers epoxy crosslinking agent immersion treatment membrane tissue material, the biology patching material obtaining is specially adapted to the reparation that hard brain/spinal meninges is damaged, after in implant into body or animal body, can keep in vivo stable in the year of June-2 does not degrade, when exceeding i.e. degraded after 2 years, guarantee also has biocompatibility when having certain mechanical property and anti-fluid seepage effect, the risk that produces post-operative complication and side effect reduces greatly, meet for the particular requirement for repairing the damaged material of hard brain/spinal meninges,
(2) hard brain/spinal meninges biology patching material preparation method of the present invention, be linking agent by selecting in propanetriol-diglycidyl-ether and/or diglycidyl ether of ethylene glycol, can not increase the cytotoxicity of this hard brain/spinal meninges biological sticking patch itself, reduce the risk that it brings side effect, state in the use the sticking patch of linking agent, though having slightly, cytotoxicity promotes, but its lifting amplitude is less, negligible, and its toxicity all, within the scope of 0 grade, can't produce adverse influence to implanted human body or animal;
(3) hard brain/spinal meninges biology patching material preparation method of the present invention, the biology patching material lipid content that is used in conjunction with acquisition by alkali lye and tensio-active agent obviously reduces, and has greatly improved security and the validity of biological sticking patch.
Brief description of the drawings
For content of the present invention is more likely to be clearly understood, below according to a particular embodiment of the invention and by reference to the accompanying drawings, the present invention is further detailed explanation, wherein
Fig. 1 is the stereoscan photograph of uncrosslinked mould material;
Fig. 2 is the stereoscan photograph of crosslinked mould material;
Fig. 3 is the control group photo of implanting uncrosslinked biological sticking patch;
Fig. 4 is the photo of implanting the test group of hard brain/spinal meninges biological sticking patch of preparing of embodiment 1;
The HE colored graph of biological sticking patch prepared by Fig. 5 embodiment of the present invention 1;
Fig. 6 is the control group that does not use hard brain/spinal meninges to repair;
Fig. 7 is the test group that uses hard brain/spinal meninges biological sticking patch prepared by embodiment 1 to repair.
Embodiment
Embodiment 1
The preparation method of the hard brain/spinal meninges biological sticking patch described in this enforcement comprises the steps:
(1) membrane tissue of raw 6 months following oxen of taking-up is starting material, cleans, and removes the part that is not suitable for processing, for subsequent use;
(2) getting membrane tissue after above-mentioned cleaning, to add mass concentration be to soak in the TritonX_100 solution of 0.1wt%, and concussion is spent the night, and to control solution temperature be 0 DEG C;
(3) getting membrane tissue after above-mentioned steps is soaked, to add concentration be to soak 2h in the sodium hydroxide solution of 5mol/L, and to control solution temperature be 15 DEG C;
(4) getting membrane tissue after above-mentioned steps is soaked, to add mass concentration be immersion treatment 1h in 1wt% superoxol;
(5) getting membrane tissue after above-mentioned steps is soaked, to add concentration be to soak 15h in 1wt% propanetriol-diglycidyl-ether;
(6) getting subsequently membrane tissue after described immersion adds pH to soak at least 5h in 5.0 phosphate buffered saline buffer;
(7) the membrane tissue lyophilize after above-mentioned steps is soaked, described lyophilize program is: product pre-freeze is first cooled to-60 DEG C and vacuumizes, heats up after following, enters sublimation stage; One time sublimation temperature rises to-25 DEG C below by-35 DEG C, and the time is no less than 6 hours; The temperature that once heats up rises to-15 DEG C by-25 DEG C, and the time is 1-3 hour; Secondary temperature elevation temperature rises to 25 DEG C by-15 DEG C, and the time is 1-3 hour.Electron beam radiation disinfection taking irradiation metering as 15KGy, to obtain final product subsequently.
Embodiment 2
The preparation method of the hard brain/spinal meninges biological sticking patch described in this enforcement comprises the steps:
(1) membrane tissue of raw 6 months following oxen of taking-up is starting material, cleans, for subsequent use;
(2) getting membrane tissue after above-mentioned cleaning, to add mass concentration be to soak in the Tween80 solution of 5wt%, and concussion is spent the night, and to control solution temperature be 25 DEG C;
(3) getting membrane tissue after above-mentioned steps is soaked, to add concentration be to soak 4h in the potassium hydroxide solution of 0.1mol/L, and to control solution temperature be 5 DEG C;
(4) getting membrane tissue after above-mentioned steps is soaked, to add mass concentration be immersion treatment 5h in 5wt% peroxide solutions;
(5) getting membrane tissue after above-mentioned steps is soaked, to add mass concentration be to soak 15h in 1wt% diglycidyl ether of ethylene glycol;
(6) getting subsequently membrane tissue after described immersion adds pH to soak at least 5h in 8.0 phosphate buffered saline buffer;
(7) the membrane tissue lyophilize after above-mentioned steps is soaked, described lyophilize program is: product pre-freeze is first cooled to-60 DEG C and vacuumizes, heats up after following, enters sublimation stage; One time sublimation temperature rises to-25 DEG C below by-35 DEG C, and the time is no less than 6 hours; The temperature that once heats up rises to-15 DEG C by-25 DEG C, and the time is at 1-3 hour; Secondary temperature elevation temperature rises to 25 DEG C by-15 DEG C, and the time is at 1-3 hour.Electron beam radiation disinfection taking irradiation metering as 25KGy, to obtain final product subsequently.
Embodiment 3
The preparation method of the hard brain/spinal meninges biological sticking patch described in this enforcement comprises the steps:
(1) membrane tissue of raw 6 months following oxen of taking-up is starting material, cleans, for subsequent use;
(2) getting membrane tissue after above-mentioned cleaning, to add mass concentration be to soak containing in TritonX_100 and Tween40 solution of 2wt%, and concussion is spent the night, and to control solution temperature be 8 DEG C;
(3) getting membrane tissue after above-mentioned steps is soaked, to add concentration be to soak 2h in the sodium hydroxide solution of 1mol/L, and to control solution temperature be 8 DEG C;
(4) getting membrane tissue after above-mentioned steps is soaked, to add mass concentration be immersion treatment 3h in 2wt% peroxide solutions;
(5) get above-mentioned steps soak after membrane tissue add mass concentration be 2wt% contain propanetriol-diglycidyl-ether and diglycidyl ether of ethylene glycol mixed cross-linker in soak 20h;
(6) getting subsequently membrane tissue after described immersion adds pH to soak at least 5h in 6.8 phosphate buffered saline buffer;
(7) the membrane tissue lyophilize after above-mentioned steps is soaked, described lyophilize program is: product pre-freeze is first cooled to-60 DEG C and vacuumizes, heats up after following, enters sublimation stage; One time sublimation temperature rises to-25 DEG C below by-35 DEG C, and the time is no less than 6 hours; The temperature that once heats up rises to-15 DEG C by-25 DEG C, and the time is at 1-3 hour; Secondary temperature elevation temperature rises to 25 DEG C by-15 DEG C, and the time is at 1-3 hour.Electron beam radiation disinfection taking irradiation metering as 30KGy, to obtain final product subsequently.
Embodiment 4
The preparation method of the hard brain/spinal meninges biological sticking patch described in this enforcement comprises the steps:
(1) membrane tissue of raw 6 months following oxen of taking-up is starting material, cleans, for subsequent use;
(2) getting membrane tissue after above-mentioned cleaning, to add mass concentration be to soak containing in TritonX_100 and Tween80 solution of 3wt%, and concussion is spent the night, and to control solution temperature be 0 DEG C;
(3) getting membrane tissue after above-mentioned steps is soaked, to add concentration be to soak 4h in the sodium hydroxide solution of 1mol/L, and to control solution temperature be 8 DEG C;
(4) getting membrane tissue after above-mentioned steps is soaked, to add mass concentration be immersion treatment 4h in 4wt% peroxide solutions;
(5) get above-mentioned steps soak after membrane tissue add mass concentration be 5wt% contain propanetriol-diglycidyl-ether and diglycidyl ether of ethylene glycol mixed cross-linker in soak 10d;
(6) getting subsequently membrane tissue after described immersion adds pH to soak at least 5h in 8.0 phosphate buffered saline buffer;
(7) the membrane tissue lyophilize after above-mentioned steps is soaked, described lyophilize program is: product pre-freeze is first cooled to-60 DEG C and vacuumizes, heats up after following, enters sublimation stage; One time sublimation temperature rises to-25 DEG C below by-35 DEG C, and the time is no less than 6 hours; The temperature that once heats up rises to-15 DEG C by-25 DEG C, and the time is at 1-3 hour; Secondary temperature elevation temperature rises to 25 DEG C by-15 DEG C, and the time is at 1-3 hour.Electron beam radiation disinfection taking irradiation metering as 10KGy, to obtain final product subsequently.
Reference examples 1
The present embodiment is identical with the preparation method of embodiment 1, and its difference is only not pass through the step of linking agent processing.
Reference examples 2
The present embodiment is identical with the preparation method of embodiment 2, and its difference is only not pass through the step of linking agent processing.
Reference examples 3
The present embodiment is identical with the preparation method of embodiment 1, and its difference is only the step of not soaking through described tensio-active agent.
Reference examples 4
The present embodiment is identical with the preparation method of embodiment 1, and its difference is only that linking agent used is glutaraldehyde cross-linking.
Effect example
Below example is investigated hard brain/spinal meninges biology patching material properties prepared by the following related embodiment of the present invention and reference examples by experiment, and then proves that hard brain/spinal meninges biological sticking patch prepared by the present invention has remarkable technique effect.
1, tear edge test
Test method: use (compression) trier that stretches, by the biology patching material of preparing according to embodiment 1-2, reference examples 1-2 method, sample is cut into the strip that specification is 1cm × 3cm, passing sample from 3-5mm place, minor face edge with 4-0 suture line, doubling suture line, apart from the about 5cm of perforation place place, suture line being tied a knot, prevent that suture line from coming off; The sample that aforesaid method is obtained, with purified water aquation 3-5 minute, one end of threading is not fixed on to the bottom of tension testing machine, threading one end is fixed on to the top of tension testing machine by hook, start to detect, until sample is torn, read the maximum value of tensile loading power, be the tear edge of this sample.The results are shown in following table 1 and Fig. 1-2.
Table 1 tear edge test result
| Test group | Tear edge (N) |
| Embodiment 1 | 13.25 |
| Embodiment 2 | 10.65 |
| Reference examples 1 | 8.56 |
| Reference examples 2 | 7.35 |
| Reference examples 4 | 10.01 |
Conclusion: upper table is three tests mean values of totally 30 flake products tear edges, embodiment 1,2 is no matter under 15KGy or the dosage of 25KGy, with respect to the situation of the reference examples 1,2 of cross-linking agent-free processing, tear edge all has obvious lifting, lifting ratio is respectively 54.79% and 44.90%, and with respect to adopting the reference examples 4 of other linking agent processing, with respect to the situation of the reference examples 1,2 of cross-linking agent-free processing, same tear edge all has obvious lifting, and lifting ratio is respectively 16.94% and 36.19%.Between the fiber forming due to the random braiding of collegen filament of membrane tissue material, there are a large amount of irregular gaps, porosity mainly depends on the structure of natural collagen(ous) fibre, linking agent can form new key and collegen filament be strained to the slight shrinkage that causes mould material between collagen, from Fig. 1-2, contrast known, before and after crosslinked, the collegen filament of biological sticking patch have obvious contraction, prove to use linking agent of the present invention to have cross-linking effect, and the collegen filament of film are had to certain tensioning function.
2, external degradation aspect
Test method: the biological sticking patch of preparing according to embodiment 1-2, reference examples 3-4 method is divided to the strip that is cut into (1-3) * (4-6) mm size, be positioned over respectively in test tube, add a certain amount of physiological saline, be placed in 37 DEG C after 1d, change one time solution, then place 3-4d, get solution and detect according to the detection method in medical device industry standard 16886, test-results sees the following form 2.
Table 2 degrading quality per-cent
| Test group | Sampling for the first time | Sampling for the second time |
| Embodiment 1 | 3.5％ | 1.4％ |
| Embodiment 2 | 6.2％ | 2.4％ |
| Reference examples 1 | 15.1％ | 7.3％ |
| Reference examples 2 | 21.8％ | 7.6％ |
| Reference examples 4 | 4.1％ | 1.8 |
Form shows to be respectively the degrading quality per-cent of 15KGy and 25KGy irradiation dose biological sticking patch above, by relatively finding out that, in the situation that having linking agent effect, the degradation speed of biological sticking patch obviously declines.
3, vivo degradation aspect
Test method: by the biological sticking patch of preparing according to embodiment 1, reference examples 1, reference examples 4 methods, the biological sticking patch original position of above-mentioned preparation is implanted in healthy rat body, described Implantation Test method is as follows: get 20 male SD rats, age in 3-4 week, body weight 180-220g, the biological sticking patch of subcutaneous implantation embodiment 1 after anesthesia, at the biological sticking patch of offside implantation reference examples 1 or reference examples 4, every material 1cm
2, choose and implant each 5 rats execution afterwards of 3 week, 12 week, 30 week, the fixing photograph of surrounding tissue paraformaldehyde of getting described implantation region.Test-results is in table 3 and Fig. 3-4, Fig. 3 is the mouse photo after 12 week of implantation of reference examples 1, Fig. 4 is the mouse photo after 12 week of implantation of embodiment 1, as can be seen from the figure, Fig. 3 is degradable, and Fig. 4 also has undegradable biological sticking patch to exist, and film form is more complete, film and the surrounding tissue situation that do not stick together, degradation time is obviously than uncrosslinked or select biological sticking patch prepared by other linking agents to extend.Test-results sees the following form 3.
Table 3 vivo degradation test-results
| Test group | 3 months | 12 months | 30 months |
| Embodiment 1 | Not degraded | Not degradable | Degradable |
| Reference examples 1 | Not degradable | Degradable | —— |
| Reference examples 4 | Not degraded | Not degradable | Not degradable |
4, cytotoxicity aspect
Test method: the biological sticking patch of preparing according to the method for embodiment 1, reference examples 1, reference examples 4 is carried out to lixiviate, prepare vat liquor; Prepare corresponding cell suspension; Discard former substratum, in 96 orifice plates, add vat liquor, negative control, positive control and the blank of proper concn, be placed in 37 DEG C of CO2gas incubator and cultivate respectively 24h, 48h.After 24h and 48h, observation of cell upgrowth situation, discards the vat liquor in 96 well culture plates, and adds the MTT solution 50 μ L of 1mg/ml.Add 96 orifice plates of MTT solution to be placed in 37 DEG C of CO2gas incubator 2 hours.After 2 hours, discard the MTT solution in 96 orifice plates, and add Virahol 100 μ L, concussion 96 orifice plate 5-10min, measure its absorbance at 492nm place.Cytotoxic detection method is the detection that the mtt assay in detecting with reference to cytotoxicity in 16886 industry standards carries out.The results are shown in following table 4,
Table 4 cell toxicity test result
| Test group | Observation period 24h | Observation period 48h |
| Embodiment 1 | 103.12 | 104.07 |
| Reference examples 1 | 100.31 | 101.25 |
| Reference examples 4 | 92.02 | 90.84 |
Upper table is three Cytotoxic mean values of test products, can be clearly seen that the product that embodiment 1 uses linking agent to prepare, cytotoxicity decreases, and all within the scope of 0 grade, down ratio than the cytotoxicity embodiment 1 of reference examples 1 is 2.80% and 2.79%, cytotoxicity changes little, illustrate and add after this linking agent, can not increase the cytotoxicity of product itself, but there is certain increase than the cytotoxicity of the cytotoxicity reference examples 4 of reference examples 1, and increase ratio is 9.01% and 11.46%, illustrate that in reference examples 4, linking agent used has certain cytotoxicity.
5, lipid content aspect
Test method: the biological sticking patch of preparing according to embodiment 1, reference examples 3 methods is tested, and determination of fat method is to formulate according to two of the Pharmacopoeias of the People's Republic of China " pancreatin fat detects ", and measurement result sees the following form 5.
Table 5 lipid content test-results
| ? | Reference examples 3 | Embodiment 1 | Lifting ratio |
| Lipid content | 0.27％ | 0.06％ | 77.77％ |
Upper table is the mean value of three test 0.1g biological sticking patch lipid contents, can be clearly seen that the embodiment of the present invention 1 is used in conjunction with the biological sticking patch of acquisition by alkali lye NaOH and surfactivity agent TrironX-100, and lipid content obviously reduces.
6, security and validity
6.1 security
Test method: biological sticking patch prepared by the embodiment of the present invention 1 carries out HE dyeing to be processed, described HE dyeing treatment step is as follows: by material with gradient alcohol descending enter water, successively clean with tap water and distilled water afterwards, after processing with 0.5% Hematorylin, clean with tap water, urge to clean after basket again with cleaning with tap water after 1% hydrochloride alcohol color separation, then with 1/400 ammoniacal liquor, rear with 80% alcohol color separation with 1% Yihong dyeing, gradient alcohol dehydration, obtains resin mounting after dimethylbenzene is transparent.The HE colored graph of biology patching material prepared by the embodiment of the present invention 1 is shown in Fig. 5, as we know from the figure, this biological sticking patch do not have cell and nucleus residual, prove that method of the present invention can effectively slough the foreign material such as the cell in mould material, safe.
6.2 validity
Test method: biological sticking patch prepared by the embodiment of the present invention 1 is implanted in healthy rabbit as test group, with do not use pachymeninx repair rabbit as a control group, described test method is as follows: after anesthesia rabbit, open rabbit skull bone, destroy endocranium, add biological sticking patch to substitute endocranium, control group is implantable bioartificial sticking patch not; Then skull bone is resetted, sew up a wound.Regularly check experimental result, test-results is shown in Fig. 6-7, Fig. 6 is the control group that does not use pachymeninx to repair, Fig. 7 is the test group photo that uses hard brain/spinal meninges biological sticking patch of preparing of the embodiment of the present invention 1, as can be seen from the figure the wound of two pictures is all fully recovered, but in Fig. 6 photo, scar is larger, and there is portion of tissue to occur downright bad phenomenon, and wound recovers good in Fig. 7 photo, and scar is less, wound is level and smooth, there is no tissue necrosis phenomenon, illustrates that repair tissue function is good and without immunological rejection.
Obviously, above-described embodiment is only for example is clearly described, and the not restriction to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here without also giving exhaustive to all embodiments.And the apparent variation of being extended out thus or variation are still among the protection domain in the invention.
Claims (10)
1. the preparation method of hard brain/spinal meninges biology patching material, it is characterized in that, comprise that getting animal membrane is organized as starting material, and be placed in turn the step that Racemic glycidol ethers epoxy crosslinking agent solution that surfactant soln, alkali lye, peroxide solutions, mass concentration are 1-5wt% and phosphate buffered saline buffer soak, and by after the described membrane tissue material lyophilize after above steps is soaked through irradiation sterilization, to obtain final product.
2. the preparation method of hard brain/spinal meninges biology patching material according to claim 1, it is characterized in that, in the step of described Racemic glycidol ethers epoxy crosslinking agent solution soaking, described linking agent is propanetriol-diglycidyl-ether and/or diglycidyl ether of ethylene glycol, and described soak time is 15h-10d.
3. the preparation method of hard brain/spinal meninges biology patching material according to claim 1 and 2, it is characterized in that, in the step that described surfactant soln soaks, the mass concentration of described tensio-active agent is 0.1-5wt%, and controls described solution temperature and be 0-25 DEG C and soak.
4. according to the preparation method of the arbitrary described hard brain/spinal meninges biology patching material of claim 1-3, it is characterized in that, in the step of described dipping by lye, the concentration of described alkali lye is 0.1-5mol/L, and controls described solution temperature and be 5-15 DEG C and soak.
5. according to the preparation method of the arbitrary described hard brain/spinal meninges biology patching material of claim 1-4, it is characterized in that, in the step that described peroxide solutions soaks, the mass concentration of described peroxide solutions is 1-5wt%.
6. according to the preparation method of the arbitrary described hard brain/spinal meninges biology patching material of claim 1-5, it is characterized in that, in the step that described phosphate buffered saline buffer soaks, the pH value of described phosphate buffered saline buffer is 5.0-8.0.
7. according to the preparation method of the arbitrary described hard brain/spinal meninges biology patching material of claim 1-6, it is characterized in that, the step of described irradiation sterilization is electron beam irradiation sterilization, and irradiation metering is 10-30KGy.
8. the hard brain/spinal meninges biology patching material preparing according to the arbitrary described method of claim 1-7.
Hard brain/spinal meninges biology patching material claimed in claim 8 for the preparation of prevent or reduce tissue adhesion's material, for repairing hard brain/spinal meninges damage that the material, treatment of impaired hard brain/spinal meninges tissue or prevention cause because of wound or tumour or damaged material, treatment or prevention because of the purposes of the damaged cerebrospinal leak causing of brain/spinal meninges, infected material firmly.
10. a Racemic glycidol ethers epoxy crosslinking agent is for the preparation of the purposes of hard brain/spinal meninges biology patching material.
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| CN201410373258.6A CN104130437B (en) | 2014-07-31 | 2014-07-31 | A kind of hard brain/spinal meninges biology patching material and its preparation method and application |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105597151A (en) * | 2016-03-09 | 2016-05-25 | 烟台正海生物科技股份有限公司 | Surgery patch and preparing method thereof |
| CN106139247A (en) * | 2015-04-17 | 2016-11-23 | 天津市赛宁生物工程技术有限公司 | A kind of preparation method of artificial hard brain (ridge) film sticking patch |
| CN107469145A (en) * | 2017-09-22 | 2017-12-15 | 北京华信佳音医疗科技发展有限责任公司 | A kind of preparation and its application of dura mater patching material |
| CN116531567A (en) * | 2023-05-09 | 2023-08-04 | 成都睿科美医疗科技有限公司 | Endometrial repair material |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4626286A (en) * | 1983-10-31 | 1986-12-02 | Schmid Laboratories, Inc. | Collagen gel and the process of making said gel |
| US5674298A (en) * | 1994-10-21 | 1997-10-07 | The Board Of Regents Of The University Of Michigan | Calcification-resistant bioprosthetic tissue and methods of making same |
| CN103272272B (en) * | 2013-05-22 | 2014-11-19 | 烟台正海生物技术有限公司 | Natural skin patch with holes and preparation method thereof |
| CN103239760B (en) * | 2013-05-22 | 2014-11-05 | 烟台正海生物技术有限公司 | Anti-adhesion surgical patch and preparation method thereof |
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| CN106139247A (en) * | 2015-04-17 | 2016-11-23 | 天津市赛宁生物工程技术有限公司 | A kind of preparation method of artificial hard brain (ridge) film sticking patch |
| CN106139247B (en) * | 2015-04-17 | 2019-11-12 | 天津市赛宁生物工程技术有限公司 | A kind of preparation method of artificial hard brain/spinal meninges sticking patch |
| CN105597151A (en) * | 2016-03-09 | 2016-05-25 | 烟台正海生物科技股份有限公司 | Surgery patch and preparing method thereof |
| CN107469145A (en) * | 2017-09-22 | 2017-12-15 | 北京华信佳音医疗科技发展有限责任公司 | A kind of preparation and its application of dura mater patching material |
| CN116531567A (en) * | 2023-05-09 | 2023-08-04 | 成都睿科美医疗科技有限公司 | Endometrial repair material |
| CN116531567B (en) * | 2023-05-09 | 2024-02-13 | 成都睿科美医疗科技有限公司 | Endometrial repair material |
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