CN114184441B - Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution - Google Patents
Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution Download PDFInfo
- Publication number
- CN114184441B CN114184441B CN202111218157.8A CN202111218157A CN114184441B CN 114184441 B CN114184441 B CN 114184441B CN 202111218157 A CN202111218157 A CN 202111218157A CN 114184441 B CN114184441 B CN 114184441B
- Authority
- CN
- China
- Prior art keywords
- leaching
- flexible
- pipe
- medical
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000002386 leaching Methods 0.000 title claims abstract description 190
- 241001465754 Metazoa Species 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000002474 experimental method Methods 0.000 title abstract description 28
- 238000011156 evaluation Methods 0.000 title abstract description 19
- 238000005070 sampling Methods 0.000 claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 238000000502 dialysis Methods 0.000 claims abstract description 21
- 238000004321 preservation Methods 0.000 claims abstract description 11
- 238000009413 insulation Methods 0.000 claims abstract description 7
- 230000009172 bursting Effects 0.000 claims abstract description 5
- 239000000463 material Substances 0.000 claims description 17
- 238000012360 testing method Methods 0.000 claims description 7
- -1 polytetrafluoroethylene Polymers 0.000 claims description 3
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 3
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 3
- 230000008569 process Effects 0.000 abstract description 7
- 230000006378 damage Effects 0.000 abstract description 5
- 239000000243 solution Substances 0.000 description 64
- 239000000047 product Substances 0.000 description 38
- 210000004027 cell Anatomy 0.000 description 19
- 230000008439 repair process Effects 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 102000008186 Collagen Human genes 0.000 description 6
- 108010035532 Collagen Proteins 0.000 description 6
- 230000009471 action Effects 0.000 description 6
- 210000000988 bone and bone Anatomy 0.000 description 6
- 229920001436 collagen Polymers 0.000 description 6
- 230000033228 biological regulation Effects 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000011010 flushing procedure Methods 0.000 description 5
- 238000007710 freezing Methods 0.000 description 5
- 230000008014 freezing Effects 0.000 description 5
- 210000005036 nerve Anatomy 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 206010070834 Sensitisation Diseases 0.000 description 4
- 238000012925 biological evaluation Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000003761 preservation solution Substances 0.000 description 4
- 230000008313 sensitization Effects 0.000 description 4
- 230000000638 stimulation Effects 0.000 description 4
- 230000002411 adverse Effects 0.000 description 3
- 210000003850 cellular structure Anatomy 0.000 description 3
- 210000004087 cornea Anatomy 0.000 description 3
- 230000006866 deterioration Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 210000000578 peripheral nerve Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000283690 Bos taurus Species 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 210000001951 dura mater Anatomy 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000012633 leachable Substances 0.000 description 2
- 238000006241 metabolic reaction Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 210000000214 mouth Anatomy 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000005653 Brownian motion process Effects 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 108010022355 Fibroins Proteins 0.000 description 1
- 206010019909 Hernia Diseases 0.000 description 1
- 229910000861 Mg alloy Inorganic materials 0.000 description 1
- 229920000954 Polyglycolide Polymers 0.000 description 1
- 229910001069 Ti alloy Inorganic materials 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 238000005537 brownian motion Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000515 collagen sponge Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 210000003516 pericardium Anatomy 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229940068918 polyethylene glycol 400 Drugs 0.000 description 1
- 239000004633 polyglycolic acid Substances 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000004876 tela submucosa Anatomy 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a preparation device and a preparation method of biological safety evaluation experiment leaching liquor of animal-derived medical equipment, wherein the device comprises a sampling tube, a flexible leaching tube, a medical dialysis pump and heat preservation equipment; the flexible leaching pipe is characterized in that the sampling pipe is cylindrical, end covers are arranged at two ends of the sampling pipe, openings are formed in the middle of the end covers, the end covers at least one end of the flexible leaching pipe are detachable, the two ends of the flexible leaching pipe are communicated with the sampling pipe through the openings in the end covers at the two ends of the sampling pipe to form a closed loop, the length of the flexible leaching pipe is matched with the amount of leaching liquid in the closed loop, so that the leaching liquid is filled in the closed loop, the sampling pipe, the flexible leaching pipe and the flexible leaching pipe are arranged in heat insulation equipment, part of the flexible leaching pipe stretches out and penetrates through the medical dialysis pump near the heat insulation equipment, and the fluctuation pressure provided by the medical dialysis pump is smaller than the bursting strength of a sample to be extracted. The leaching solution is prepared by the device, and medical instruments in the leaching solution are not easy to damage in the leaching process.
Description
Technical Field
The invention belongs to the field of biological safety evaluation of medical instruments, and particularly relates to a preparation device and a preparation method of biological safety evaluation experiment leaching liquor of animal-derived, in particular living cell-containing animal-derived medical instruments.
Background
The animal-derived medical apparatus mainly refers to the medical apparatus which is prepared from animal tissues by processing and preparing tissues for human body. Because the sources, structures, components and the like of the raw materials are close to human tissues, compared with medical devices prepared from other raw materials (natural polymers such as chitosan, silk fibroin and the like, organic matters such as titanium alloy, magnesium alloy metal, polylactic acid, polyglycolic acid and the like), the medical device has better tissue compatibility and biological safety. The innovation of the animal-derived medical instrument is high, and the animal-derived medical instrument is a novel medical instrument which is developed in recent years, and the raw materials, the manufacturing process, the sterilization modes and the like are various, wherein animal species used for taking raw materials comprise pigs, cows, sheep and the like, tissue organs used for taking materials comprise cornea, pericardium, peritoneum, dermal tissue, small intestine submucosa, tendon, nerve, bone tissue and the like, and the animal-derived medical instrument which has acquired the registration of the medical instrument now comprises artificial cornea, artificial dura mater, artificial skin, oral cavity or tissue repair membrane, collagen related products, artificial nerve repair materials, artificial bones and the like. Such devices are not only of different types, but also of very different morphologies.
Most of the film products belong to film sheets, and the difference between thickness and texture is large; collagen products are generally spongy and have a certain pore morphology; the appearance of the artificial nerve repair material is similar to that of a normal nerve; the artificial bone comprises bone powder, bone granule, and bone strip with certain shape.
In order to improve the performance and market competitiveness of some animal-derived medical devices, when new products are designed and developed or original products are updated, additional functions such as antibiosis and bacteriostasis, growth promotion, suture-free and the like are added through some special methods, and one method which is at present more sophisticated is to compound living cells to promote the functions of the products, improve the histocompatibility and biosafety of the products and promote the repair and regeneration of organism tissues.
There are two ways of evaluating the biosafety of medical instruments: one is to directly select the finished product of the medical apparatus or the final sample with the same treatment mode as the finished product for inspection; the other is to prepare a leaching solution from the above sample and then examine the biological safety of the leaching solution. Biological safety evaluation experimental sample leach preparation general reference GB/T16886.12-2017 medical device biology evaluation part 12: the sample preparation and reference materials define the needed proportion of the preparation of the product leaching liquid with different forms and thicknesses and four corresponding leaching conditions, and the method can be almost suitable for all traditional medical appliance products and most newly-emerging animal-derived medical appliances with higher innovation. The detailed leaching conditions are as follows:
1)、(37±1)℃(72±2)h；
2)、(50±2)℃(72±2)h；
3)、(70±2)℃(24±2)h；
2)、(121±2)℃(1±0.1)h；
the standard surface area and leach liquor volume ratios are as follows:
the leaching solution is generally classified into a polar leaching solution, a nonpolar leaching solution, and other leaching solutions. The polar leach liquor contains mainly: water, physiological saline and serum-free medium; the nonpolar leaching solution mainly comprises fresh refined vegetable oil which accords with the quality regulation of pharmacopoeia of various countries, such as cotton seed oil or sesame oil and the like; other leaching solutions mainly comprise ethanol/water, ethanol/physiological saline, polyethylene glycol 400 (diluted to physiological osmotic pressure), dimethyl sulfoxide, serum-containing culture medium and the like.
The sample leaching solution preparation method is generally as follows:
mixing the sample and the leaching solution according to the recommended ratio of the standard, placing the mixture into a glass bottle with a sealing cover, and placing the glass bottle into a constant-temperature shaking table to stir for a period of time, so that leachables such as a release agent, a plasticizer, other auxiliary agents, a coating and the like in the production process of the sample and degradation products of the sample and packaging materials and the like in the preservation process are fully dissolved into the extracting solution.
The preparation method of the sample leaching solution is expected to fully extract soluble substances in the sample without damaging the sample. The test requires that the detected sample is representative, the whole experimental result of the product can be truly reflected, and if the detected sample is metabolized or degraded or living cells fall off or die in the preparation process of the leaching solution, the finally obtained leaching solution cannot represent the actual condition of the detected sample, and the detection result is also greatly wrong.
The preparation method of the sample leaching solution has the following defects:
the liquid and air coexist in the leaching container, the sample can not be ensured to be always in the leaching liquid in the stirring process of the constant-temperature shaking table, and the combined action of the gas and the liquid can easily cause the destruction of animal tissues or the flushing of surface cells. During practice, tissue debris is often even found in suspension in the leach liquor, adversely affecting the test results.
In addition, although the above leaching conditions are applicable to most medical appliance products and have universality, with the continuous emergence of the current innovative medical appliances, for example, for collagen extraction products, the raw materials are living animals such as pigs, cattle and sheep, the normal body temperature is lower than 40 ℃, compared with the preparation conditions of the leaching solution, the leaching solution is obviously unsuitable for leaching products mainly containing collagen at the temperature of 50 ℃, 70 ℃ and 121 ℃, and the leaching conditions of 37 ℃ and 72 hours are in vitro, so that the products have deterioration reaction in the environment of the leaching solution. For example, for more specific living cell-containing animal-derived medical devices, the microenvironment of the device when it is implanted into a human body is 37 ℃, and if the leaching condition of 37 ℃ is selected, the living cell components are already subjected to metabolic reaction after 72 hours of leaching according to the above standard. The leaching solution prepared from the products subjected to metabolic reaction in the leaching solution preparation process cannot reflect the actual situation of the performance indexes of the products, and is highly likely to cause incorrect test results.
In summary, the conventional standard leaching devices, leaching conditions, and the like have poor applicability to animal-derived medical devices, particularly to animal-derived medical devices containing living cells, and the obtained biological safety evaluation results of the leaching solution cannot reflect the actual situation of the product.
Disclosure of Invention
The invention aims to provide a preparation device of biological safety evaluation experiment leaching liquor of animal-derived medical equipment, the leaching liquor is prepared by the device, medical equipment in the leaching liquor is not damaged easily in the leaching process, and the obtained biological safety evaluation result of the sample leaching liquor can better reflect the real situation of a product.
The first invention is realized by the following technical scheme: the preparation device of the experimental leaching solution for evaluating the biological safety of the animal-derived medical instrument comprises a sampling tube, a flexible leaching tube, a medical dialysis pump and heat preservation equipment;
the flexible leaching pipe is characterized in that the sampling pipe is cylindrical, end covers are arranged at two ends of the sampling pipe, openings are formed in the middle of the end covers, the end covers at least one end of the flexible leaching pipe are detachable, the two ends of the flexible leaching pipe are communicated with the sampling pipe through the openings in the end covers at the two ends of the flexible leaching pipe to form a closed loop, the length of the flexible leaching pipe is matched with the amount of leaching liquid in the closed loop, so that the leaching liquid is filled in the closed loop, the sampling pipe and the flexible leaching pipe are arranged in heat insulation equipment, part of the flexible leaching pipe extends out and penetrates through the medical dialysis pump near the heat insulation equipment, the fluctuation pressure provided by the medical dialysis pump is smaller than the bursting strength of a sample to be extracted, and the bursting strength refers to the technical requirements of products provided by a sample manufacturing enterprise.
The sample adding pipe is mainly used for placing and positioning a sample, the flexible leaching pipe is mainly used for storing leaching liquid and circularly flushing the sample in the sample adding pipe by the leaching liquid therein under the action of the medical dialysis pump, the medical dialysis pump is mainly used for providing fluctuating pressure in sample leaching equipment, and the heat preservation equipment is mainly used for keeping the temperature, in particular to providing a low-temperature environment of less than 37 ℃ such as 4-8 ℃. The length of the flexible leaching tube is intercepted according to the total amount of leaching liquid, so that the leaching liquid can be filled in the inner cavities of the sampling tube and the flexible leaching tube, the uniform and stable inner cavity pressure is ensured when the medical dialysis pump pressurizes the flexible leaching tube, and the unstable condition caused by the gas-liquid combined action in the sample leaching process is avoided.
When the device is used for preparing the sample leaching solution, air is not contained in a closed loop, the pressure is uniform, and the leachable substances and other components in the sample can be well extracted under the condition of smaller fluctuation pressure in a matched circulating flushing mode, so that the sample leaching solution is fully leached when the device is used for preparing the sample leaching solution, medical equipment in the sample leaching solution is not easy to damage, and the biological safety evaluation result of the obtained sample leaching solution can be better than the real condition of a reaction product. In addition, the lower temperature environment with the same shelf life preservation temperature condition of the sample can avoid the metabolism, degradation or deterioration of the sample.
The frequency range of fluctuation of the medical dialysis pump is recommended to be 60 times/min-120 times/min.
In order to avoid the influence of the material of the device on the leaching liquid, the sampling tube and the flexible leaching tube in the device are both made of polytetrafluoroethylene serving as inert materials.
The second object of the invention is to provide a method for preparing biological safety evaluation experiment leaching liquor of animal-derived medical equipment by using the device.
As a preferable scheme: leaching temperature: 4-8 ℃; frequency of fluctuation: 60 times/min-120 times/min; leaching time: 1-30d.
The leaching solution for leaching should be pre-cooled to a required temperature range when being added, so that potential changes of product performance caused by temperature changes are avoided, and the final sample leaching solution result is influenced.
Compared with the prior art, the invention has the following beneficial effects:
1) In the prior art, the dissolution and precipitation of soluble substances in a sample are generally promoted by shaking a container, gas and liquid coexist in the container, the pressure of liquid on the sample is large and uneven in the shaking process, and the sample is easy to damage;
2) The environmental temperature is set to be 4-8 ℃ and is consistent with the preservation temperature of the product specification, so that when the animal-derived medical instrument, particularly the leaching solution of the living cell-containing animal-derived medical instrument is prepared by the method, the conditions of metabolism, degradation, deterioration and the like of the product are avoided, and the adverse effect of improper conditions in the preparation process of the leaching solution on the evaluation of the product result is reduced;
3) The leaching conditions such as the leaching temperature, the leaching time, the fluctuation frequency and the like are verified by comparison tests, the leaching solution results are stable and reliable, and the method can be used as a standard method for preparing the leaching solution of animal-derived medical devices and living cell-containing animal-derived medical devices.
Drawings
FIG. 1 is a schematic structural view of a preferred embodiment of a device for preparing a biological safety evaluation test leaching solution for an animal-derived medical device according to the present invention;
FIG. 2 is a schematic illustration of the connection of the loading tube and flexible leaching tube of FIG. 1.
Detailed Description
The invention is further described in detail below in connection with specific embodiments and with the accompanying drawings.
The preparation device of the experimental leaching solution for evaluating the biological safety of the animal-derived medical instrument in the embodiment is shown in fig. 1, and comprises a sampling tube 1, a flexible leaching tube 2, a medical dialysis pump 3 and a low-temperature freezing cradle 4. As shown in fig. 2, the sampling tube 1 is cylindrical, two ends are provided with end caps 11, the middle of the end caps 11 is provided with openings 111, and two ends of the flexible leaching tube 2 are communicated with the sampling tube 1 through the openings 111 on the end caps 11 at two ends of the sampling tube (the flexible leaching tube 2 is specifically sleeved on a cylindrical open-pore tube on the end caps 11), so as to form a closed loop. As shown in fig. 1, the length of the flexible leaching tube 2 is adapted to the amount of leaching liquid in the closed circuit so that the closed circuit is filled with leaching liquid. The medical dialysis pump 3 is arranged near the low-temperature freezing cradle 4. After the sampling tube 1 is tightly connected with the flexible leaching tube 2, other parts except the part which needs to be connected with the medical dialysis pump 3 are immersed in a temperature control area of a low-temperature freezing cradle at the temperature of 4-8 ℃.
The sampling tube 1 mainly plays a role of placing and positioning a sample, the flexible leaching tube 2 mainly plays a role of storing leaching liquid, and the sample in the sampling tube 1 is circularly washed by the leaching liquid under the action of the medical dialysis pump 3, the medical dialysis pump 3 is mainly used for providing the fluctuation pressure in sample leaching equipment, the fluctuation pressure provided by the medical dialysis pump 3 is less than the bursting strength of the sample to be extracted, and the low-temperature freezing shaking table 4 is only used for providing a low-temperature environment and can be replaced by other equipment capable of providing the same temperature.
The components of the product obtained after the extraction of the collagen are mainly different types of collagen, and the product is usually stored in a refrigerating way at the temperature of 4-8 ℃. The preservation temperature of the animal-derived medical device containing the preservation solution is also generally between 4 ℃ and 8 ℃, the shelf life of the product is 1-3 years, and certain cell components of the product containing living cells cannot activate metabolism within the temperature range. Comprehensively considering, the experimental conditions of this embodiment select the leaching temperature range to be 4-8 ℃, that is, the working temperature of the low-temperature freezing cradle 4 is set to be 4-8 ℃.
The medical dialysis pump 3 is set as follows: the liquid fluctuation frequency in the leaching pipe is 60 times/min-120 times/min. In the experiment, the temperature of the leaching solution is reduced, the Brownian motion of water molecules is reduced, and meanwhile, the possibility of damaging a sample is high, so that the fluctuation frequency of the leaching solution is limited to 60 times/min-120 times/min, and the proper fluctuation frequency is selected according to the properties of the sample.
The whole leaching period is 1-30 days, and the special sample can properly prolong the leaching period to achieve the effect of full leaching. Specifically, the selection of the properties of the sample to be tested is performed.
In order to avoid the influence of the device material on the leaching solution, the sampling tube 1 and the flexible leaching tube 2 in the device are made of polytetrafluoroethylene serving as inert materials.
As shown in FIG. 1, in this embodiment, the nozzles at the two ends of the sample adding tube 1 are matched with the diameter of the leaching tube 2, and they can be firmly connected without leakage. The middle tube body of the sampling tube 1 is connected with end covers 11 on two sides through threads, a sample can be put in or taken out after the sampling tube is opened, and the sampling tube is pressure-resistant and has no leakage after being tightly covered.
The sample adding tube 1 is spirally opened, the sample is conveniently and rapidly put in and taken out, the sample adding tube 1 and the flexible leaching tube 2 are directly inserted and connected, the tightness is good, the risks of leakage and pollution are avoided, and the aseptic operation requirement of the medical instrument during biological safety evaluation leaching solution preparation is conveniently met.
Under the preservation condition of the medical equipment product containing living cells and animal sources, the leaching effect is promoted by the cyclic flushing action of the leaching solution, so that the product is leached more effectively under the gentle and uniform flushing action, and the leaching solution with smaller influence on the biological safety evaluation result is obtained. The device and the method can ensure that the products are not deteriorated, metabolized, degraded and the like during the preparation of the animal-derived medical instrument and the leaching solution of the living cell-containing animal-derived medical instrument, and can reduce the adverse effect of improper conditions in the preparation process of the leaching solution on the evaluation of the product results.
The biological safety evaluation experiment of the medical instrument is necessary data for registering and declaring most medical instruments, so the device and the method have great application value.
Examples of the preparation of biological safety evaluation test leaching solutions for animal-derived medical devices using the above apparatus are as follows:
example 1
The adding proportion of the leaching solution is in accordance with the regulation of the 12 th part of biological evaluation of GB/T16886.12-2017 medical instruments, animal-derived medical instruments such as hernia repair materials and the like have good toughness, the thickness is usually about 0.2mm, the thickness is smaller than 0.5mm, the film sample is dried and stored without preservation solution, and the film sample is prepared according to the thickness of 6cm 2 Leaching solution is added according to the ratio of/mL, and leaching temperature is as follows: the temperature is 4-8 ℃, the fluctuation frequency is 60-80 times/min, and the leaching time is 1-3 days.
Examples-comparative experiments
The comparison experiment alone increased the fluctuation frequency to 120 times/min or the leaching time to 7 days, and the increase of both the oscillation frequency and time did not change the results of the leaching solution biosafety experiment (sensitization, stimulation, etc.) under the above conditions.
A film sample of less than 0.5mm, 6cm, is described 2 The leaching solution is added in the ratio of/mL, the fluctuation frequency is 60-80 times/min, the leaching time is 1-3 days, and the purpose of full leaching can be achieved.
Example two
The adding proportion of the leaching solution is in accordance with the regulation of the 12 th part of GB/T16886.12-2017 medical instrument biological evaluation, the thickness of the diaphragm-shaped sample of the animal-derived medical instrument such as an oral cavity repairing film, an artificial dura mater and the like is more than 0.5mm, and the diaphragm-shaped sample is preserved under the condition of a preservation solutionThe material has soft texture, relatively strict leaching condition, and the leaching rate is 3cm 2 Leaching solution is added according to the ratio of/mL, and leaching temperature is as follows: the temperature is 4-8 ℃, the fluctuation frequency is 80-100 times/min, and the leaching time is 3-5 days.
Example two comparative experiments
The comparison experiment alone increased the fluctuation frequency to 120 times/min or the leaching time to 7 days, and the increase of both the oscillation frequency and time did not change the results of the leaching solution biosafety experiment (sensitization, stimulation, etc.) under the above conditions.
Illustrating that the thickness of the film sample is larger than 0.5mm, and the film sample is 3cm after preservation of preservation solution 2 The leaching solution is added in the ratio of/mL, the fluctuation frequency is 80-100 times/min, the leaching time is 3-5 days, and the purpose of full leaching can be achieved.
Example III
The adding proportion of the leaching solution is in accordance with the regulation of the 12 th part of biological evaluation of GB/T16886.12-2017 medical instruments, such as peripheral nerve repair materials and the like, the animal-derived medical instruments have solid forms with the diameters of usually 1-5mm, the appearance of which are close to that of normal nerves, and cylindrical samples with the thicknesses of more than 0.5mm, are stored under the condition of normal preservation solution, the texture of the materials is tough, the leaching condition is stricter, and the thickness of the materials is more than 3cm 2 Leaching solution is added according to the ratio of/mL, and leaching temperature is as follows: the temperature is 4-8 ℃, the fluctuation frequency is 100-120 times/min, and the leaching time is 7-15 days.
Example three comparative experiments
The comparison experiment alone increased the fluctuation frequency to 180 times/min (fluctuation frequency is frequently used for preparing the peripheral nerve repair material) or the leaching time to 30 days (fluctuation time is frequently used for preparing the peripheral nerve repair material), and the increase of the oscillation frequency and the time to the above conditions did not change the results of the leaching solution biosafety experiment (sensitization, stimulation and other experiments).
The thickness of the thick film slice sample is larger than 0.5mm, the leaching solution is added in the proportion of 3cm < 2 >/mL, the fluctuation frequency is 100-120 times/min, and the leaching time is 7-15, so that the purpose of full leaching can be achieved.
Example IV
The adding proportion of the leaching solution is in accordance with the regulation of the 12 th part of biological evaluation of GB/T16886.12-2017 medical equipment, the corresponding leaching solution proportion is selected by the animal-derived medical equipment such as living cell-containing human body repair materials and the like according to the morphological characteristics of the product, and the storage condition of the product is more strict due to the very strict requirement of living cell survival conditions, the shelf life is shorter, for example, the shelf life of the domestic first living cell-containing animal-derived artificial skin is only 30 days, and in order to avoid the influence of the product after living cell metabolism or death in the leaching process on the result of a leaching solution experiment, the leaching temperature is as follows: the leaching time is set to 3-7d at 4-8 ℃ and the fluctuation frequency is 60 times/min.
Example four comparative experiment one
Increasing the frequency of fluctuation to above 120 times/min in the comparative experiment obviously affects the adhesion of living cells, so that no recommendation is made, the frequency of fluctuation is singly increased to 80 times/min or the leaching time is increased to 30 days, and the results of the biological safety experiment (sensitization, stimulation and the like) of the leaching solution are not changed under the conditions.
The comparison shows that the leaching time of the animal-derived medical devices such as the living cell-containing human body repair materials is set to 3-7 days, the fluctuation frequency is 60 times/min, and the purpose of full leaching can be achieved.
In addition, compared with the experimental method in the standard, by taking artificial cornea and collagen sponge products as examples, the leaching solution has no turbidity, light transmittance change and the like after the preparation of the leaching solution is completed, and the method has less influence on the sample than the standard leaching method.
Example four comparative experiment two
Compared with the common leaching experimental method listed in the standard, the comparison experiment is mainly compared with the common leaching experimental method, and compared with the common experimental method, the microscopic detection shows that the number and the morphological morphology of the cells attached to the surface of the device are not obviously changed or abnormal after leaching by the method provided by the invention, which indicates that no obvious cell detachment or destruction phenomenon exists. The common method has obviously changed the product form, obviously reduced the number of living cells attached to the surface of the instrument, and the leaching solution can also find the fallen cell components during microscopic examination.
Claims (7)
1. The preparation device of the experimental leaching solution for evaluating the biological safety of the animal-derived medical instrument is characterized by comprising a sampling tube, a flexible leaching tube, a medical dialysis pump and heat preservation equipment;
the flexible leaching pipe is characterized in that the sampling pipe is cylindrical, end covers are arranged at two ends of the sampling pipe, openings are formed in the middle of the end covers, the end covers at least one end of the flexible leaching pipe are detachable, the two ends of the flexible leaching pipe are communicated with the sampling pipe through the openings in the end covers at the two ends of the sampling pipe to form a closed loop, the length of the flexible leaching pipe is matched with the amount of leaching liquid in the closed loop, so that the leaching liquid is filled in the closed loop, the sampling pipe, the flexible leaching pipe and the flexible leaching pipe are arranged in heat insulation equipment, part of the flexible leaching pipe stretches out and penetrates through the medical dialysis pump near the heat insulation equipment, and the fluctuation pressure provided by the medical dialysis pump is smaller than the bursting strength of a sample to be extracted.
2. The apparatus of claim 1, wherein the thermal insulation device is configured to provide a low temperature environment of 4-8 ℃.
3. The preparation device according to claim 1 or 2, characterized in that the frequency of fluctuation of the medical dialysis pump ranges from 60 times/min to 120 times/min.
4. A preparation device according to claim 3, wherein the sample addition tube and the flexible leaching tube are made of polytetrafluoroethylene, which is an inert material.
5. A method of preparing an animal derived medical device biosafety assessment test leach solution using the apparatus of any one of claims 1-4.
6. The method of claim 5, wherein the leaching temperature: 4-8 ℃; frequency of fluctuation: 60 times/min-120 times/min; leaching time: 1-30d.
7. The method according to claim 6, wherein the leaching solution for leaching is added pre-cooled to the desired temperature range, i.e. to 4-8 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111218157.8A CN114184441B (en) | 2021-10-19 | 2021-10-19 | Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111218157.8A CN114184441B (en) | 2021-10-19 | 2021-10-19 | Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114184441A CN114184441A (en) | 2022-03-15 |
| CN114184441B true CN114184441B (en) | 2024-04-05 |
Family
ID=80539553
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111218157.8A Active CN114184441B (en) | 2021-10-19 | 2021-10-19 | Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114184441B (en) |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5528923A (en) * | 1992-12-21 | 1996-06-25 | Ledez; Kenneth M. | Gas amount and solubility investigation apparatus |
| CN102156188A (en) * | 2011-02-10 | 2011-08-17 | 云南烟草科学研究院 | Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper |
| CN104237230A (en) * | 2014-10-14 | 2014-12-24 | 厦门奥亚仪器有限公司 | Quick melatonin detection method and operation utensil assembly thereof |
| CN204043972U (en) * | 2014-03-07 | 2014-12-24 | 中华人民共和国四川出入境检验检疫局 | A kind of to granulated samples carrying out the chemical detection liquid extraction tube of lixiviate |
| CN104931437A (en) * | 2015-05-28 | 2015-09-23 | 中国林业科学研究院热带林业实验中心 | Method for detecting contents of potassium, calcium and magnesium in plant |
| CN107367597A (en) * | 2017-08-15 | 2017-11-21 | 上海市环境科学研究院 | A kind of heavy-metal contaminated soil/waste stabilization product diafiltration leaching equipment |
| CN110170180A (en) * | 2019-04-18 | 2019-08-27 | 广东省测试分析研究所（中国广州分析测试中心） | A kind of extracting at constant temperature/extraction Suction filtration device of solid waste/heavy metal in soil Leaching |
| CN209741233U (en) * | 2018-12-29 | 2019-12-06 | 紫金矿业集团股份有限公司 | Expansion device for biological column immersion test |
| CN110987580A (en) * | 2019-12-24 | 2020-04-10 | 四川西晨智慧农业科技有限公司 | Device and method for extracting biological effective state of heavy metal in soil |
| CN211057199U (en) * | 2019-10-31 | 2020-07-21 | 万宝矿产有限公司 | Copper ore column leaching-extraction linkage test system |
| WO2020248859A1 (en) * | 2019-06-11 | 2020-12-17 | 深圳市药品检验研究院（深圳市医疗器械检测中心） | Method for limiting extraction of medical device |
| CN112500932A (en) * | 2020-12-14 | 2021-03-16 | 云南中烟新材料科技有限公司 | Device and method for improving extraction efficiency and quality of rhizome-type spice plants |
| CN113503906A (en) * | 2021-08-11 | 2021-10-15 | 清华大学 | In-vitro circulation experiment table and method for medical implant intervention body thrombosis |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7326564B2 (en) * | 2001-02-20 | 2008-02-05 | St. Jude Medical, Inc. | Flow system for medical device evaluation and production |
| US7476371B2 (en) * | 2002-02-14 | 2009-01-13 | Phibro-Tech, Inc. | Dissolution of copper metal in aqueous alkanolamine to form copper containing aqueous solution |
| BE1025528B1 (en) * | 2017-09-06 | 2019-04-05 | Universite De Liege Établissement Public De La Communauté Française De Belgique | NON-DESTRUCTIVE SAMPLE DEVICE FOR EXTRACTING MARKERS |
-
2021
- 2021-10-19 CN CN202111218157.8A patent/CN114184441B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5528923A (en) * | 1992-12-21 | 1996-06-25 | Ledez; Kenneth M. | Gas amount and solubility investigation apparatus |
| CN102156188A (en) * | 2011-02-10 | 2011-08-17 | 云南烟草科学研究院 | Neutral erythrocyte toxicity testing method for biological evaluation of cigarette tipping paper |
| CN204043972U (en) * | 2014-03-07 | 2014-12-24 | 中华人民共和国四川出入境检验检疫局 | A kind of to granulated samples carrying out the chemical detection liquid extraction tube of lixiviate |
| CN104237230A (en) * | 2014-10-14 | 2014-12-24 | 厦门奥亚仪器有限公司 | Quick melatonin detection method and operation utensil assembly thereof |
| CN104931437A (en) * | 2015-05-28 | 2015-09-23 | 中国林业科学研究院热带林业实验中心 | Method for detecting contents of potassium, calcium and magnesium in plant |
| CN107367597A (en) * | 2017-08-15 | 2017-11-21 | 上海市环境科学研究院 | A kind of heavy-metal contaminated soil/waste stabilization product diafiltration leaching equipment |
| CN209741233U (en) * | 2018-12-29 | 2019-12-06 | 紫金矿业集团股份有限公司 | Expansion device for biological column immersion test |
| CN110170180A (en) * | 2019-04-18 | 2019-08-27 | 广东省测试分析研究所（中国广州分析测试中心） | A kind of extracting at constant temperature/extraction Suction filtration device of solid waste/heavy metal in soil Leaching |
| WO2020248859A1 (en) * | 2019-06-11 | 2020-12-17 | 深圳市药品检验研究院（深圳市医疗器械检测中心） | Method for limiting extraction of medical device |
| CN211057199U (en) * | 2019-10-31 | 2020-07-21 | 万宝矿产有限公司 | Copper ore column leaching-extraction linkage test system |
| CN110987580A (en) * | 2019-12-24 | 2020-04-10 | 四川西晨智慧农业科技有限公司 | Device and method for extracting biological effective state of heavy metal in soil |
| CN112500932A (en) * | 2020-12-14 | 2021-03-16 | 云南中烟新材料科技有限公司 | Device and method for improving extraction efficiency and quality of rhizome-type spice plants |
| CN113503906A (en) * | 2021-08-11 | 2021-10-15 | 清华大学 | In-vitro circulation experiment table and method for medical implant intervention body thrombosis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114184441A (en) | 2022-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1307246B1 (en) | Biomaterial including animal corneal tissue | |
| Geerts et al. | Nondestructive methods for monitoring cell removal during rat liver decellularization | |
| CN111084904B (en) | Cell removing method of tissue engineering scaffold | |
| CN104886038A (en) | Method for producing animal specimen | |
| CN105802252B (en) | A kind of collagen-modified method and use modified collagen albumen made from the method | |
| CN114184441B (en) | Preparation device and method of animal-derived medical instrument biosafety evaluation experiment leaching solution | |
| Isik et al. | 3D Printing of Extracellular Matrix‐Based Multicomponent, All‐Natural, Highly Elastic, and Functional Materials toward Vascular Tissue Engineering | |
| CN105251061B (en) | A kind of biodegradable post-operation adhesion preventing composite membrane and preparation method thereof | |
| CN102895702A (en) | Composite manual bile duct and preparation method thereof | |
| CN111330076B (en) | Cell removing device of tissue engineering bracket | |
| US20230320709A1 (en) | Devices and methods for umbilical cord processing | |
| CN107427608A (en) | Utilize the artificial biological film and its manufacture method of silk matrix | |
| Ozturk et al. | Use of diced cartilage grafts wrapped with amniotic membrane in soft tissue augmentation: experimental study | |
| JP5763790B2 (en) | Biological transplant material derived from mammalian cartilage tissue | |
| Shirakigawa et al. | Decellularization of liver and organogenesis in rats | |
| EP3919607B1 (en) | Multi-compartment container for culturing and surface multiplication of human autologous fibroblasts on a membrane carrier | |
| CN205087344U (en) | Biological tissue freezes and deposits container | |
| WO2021006335A1 (en) | Connective-tissue body and method for producing same | |
| KR102171320B1 (en) | A method for decellularizing of a tracheal mucosa tissue | |
| RU2797632C1 (en) | Method of manufacturing a vascular graft from the vein of the human umbilical cord | |
| CN105963784B (en) | Method and reagent for recovering residual stress and residual strain of decellularized blood vessel | |
| CN106794282B (en) | Vascular patch and its manufacturing method based on silk cocoon | |
| KR102577231B1 (en) | Intestinal extracellular matrix-derived scaffold for culture and transplantation of intestinal organoid and preparing method thereof | |
| CN109370982A (en) | A kind of chick embryo extract and its preparation and application | |
| CN114557336B (en) | Tissue treatment fluid capable of improving primary separation quantity and activity of tissues as well as preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |