CN110692647A - Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering - Google Patents
Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering Download PDFInfo
- Publication number
- CN110692647A CN110692647A CN201910371621.3A CN201910371621A CN110692647A CN 110692647 A CN110692647 A CN 110692647A CN 201910371621 A CN201910371621 A CN 201910371621A CN 110692647 A CN110692647 A CN 110692647A
- Authority
- CN
- China
- Prior art keywords
- sterilization
- tissue engineering
- allogeneic
- lysozyme
- solution
- 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.)
- Pending
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
Abstract
The invention discloses a sterilization liquid and a sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering. The sterilizing liquid and the method can effectively carry out final sterilization on products under the condition of not damaging active cells, related factor proteins and matrix structures in the tissue engineering medical instruments. Finally, the tissue engineering medical instrument is matched with aseptic packaging to finally reach an aseptic state.
Description
Technical Field
The invention relates to the technical field of medical product sterilization, in particular to a sterilization solution and a sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering.
Background
The Non-destination liquid sterilization method is mainly applied to the terminal sterilization of part of allogeneic and xenogeneic tissue engineering medical instruments. As a disposable medical device, it is ultimately intended to be sterile for clinical use. At present, the methodFor such single-use sterile medical device products, there are generally two preparation methods. The first method comprises the following steps: terminal sterilization refers to sterilization of the product after preparation and then aseptic packaging, or sterilization after packaging (e.g., irradiation sterilization). And the second method comprises the following steps: sterile operation method: the method is characterized in that raw and auxiliary materials, station appliances, equipment and the like of the product are sterilized and then are operated in an aseptic environment until the product is packaged. Both methods finally achieve a sterilization assurance level of 10-6. In the clinic, almost all disposable sterile medical devices adopt the first mode. The second sterilization mode is relatively high in cost and difficult to control, and only a few products (such as culture media and the like) are adopted. The Non-destination liquid sterilization rule belongs to the first sterilization mode.
For allogeneic and xenogeneic tissues, it is a basic raw material for many tissue engineered medical devices. The material can carry various fungi and viruses, and the material mainly comprises various cells and matrixes, and is also a natural culture medium. If storage is problematic, a large amount of bacteria or viruses will grow on the material in a short period of time. The allogenic or xenogenic tissue usually used as raw material has large volume and high storage requirement, and can not be directly and effectively sterilized before use. Thus, the final aseptic operation cannot be achieved. If the first terminal sterilization mode (such as irradiation, ethylene oxide, dry heat, moist heat, ethanol, etc.) is used to sterilize the product, the structure of the tissue engineering product is seriously damaged, and more importantly, the microenvironment in which the cells grow is destroyed. After the tissue engineering product is implanted into a human body, cells of a receptor need to be attracted to grow in, and capillary crawling coverage provides nutrition for survival. If the sterilization mode of irradiation, ethylene oxide, dry heat, moist heat, ethanol and the like is adopted, the sterilization also destroys the activity of cells and the microenvironment on which the cells live at the same time: we have found that human and animal tissues contain a large number of factors or proteins that facilitate wound repair, growth, induction, infection resistance, etc. Such as: human cortical growth factor (HEGP), osteoinductive growth factor (BMP), intravascular cortical growth factor (VEGF), and the like. These substances can induce or guide cells in the affected area for repair treatment of the damaged portion. At the same time, a plurality of factors and proteins can control the effect after tissue repair, and finally, the state which is almost the same as that before damage can be achieved. This is extremely important for repair of many injuries. Such as: skin grafting after burn, plastic cosmetology, etc.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a sterilization solution and a sterilization method suitable for allogeneic and xenogeneic tissues and tissue engineering medical instruments, and the sterilization solution and the method can effectively carry out final sterilization on products under the condition of not damaging active cells, related factor proteins and matrix structures in the tissue engineering medical instruments. Finally, the tissue engineering medical instrument is matched with aseptic packaging to finally reach an aseptic state.
The invention aims to provide a sterilizing solution suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering, which comprises lysozyme and antibiotics.
The medical apparatus of the allogeneic and xenogeneic tissues and the tissue engineering is a functional product which has the functions of the biological structure, the intercellular substance and part of growth factors of biological tissue materials, does not retain the cell activity and does not obtain the cell functionality. Including allogeneic and xenogeneic tissue material.
The method of the present invention is applicable to most conventional tissue engineering medical devices with a wide range of applications, but in order to exert better sterilization advantages and for cost reasons, it is preferred that the allogeneic and xenogeneic tissue and tissue engineering medical devices:
(1) functional products which are soluble in water, lipids or/and alcohols are excluded; such as subcutaneous fat layer, vitreous humor:
(2) materials that do not contain moisture in the final product are excluded; such as: the chitin material is specifically as follows: hyaluronic acid freeze-dried powder and adsorption test paper;
(3) does not include some tissue engineering materials with strong adsorption; such as: biological sponge materials, such as PVA sponge and polyurethane sponge;
(4) does not comprise a matrix material needing the growth of the attached cells and a tissue engineering medical apparatus with drug loading implantation; such as: a cardiovascular stent;
(5) materials for cardiac pacemakers that do not include bone graft materials for functionality; such as: artificial hearts, and the like.
Further, the allogeneic and xenogeneic tissues and tissue engineering medical devices of the present invention may be selected from allogeneic or xenogeneic tissues other than those mentioned above, including but not limited to skin, bone, cartilage, tendon, nerve tissue, blood vessels, cardiac valves, pericardium, amnion, umbilical cord tissue, etc.
Furthermore, the lysozyme is selected from one or more of animal-derived lysozyme, plant-derived lysozyme and microorganism-derived lysozyme. Wherein the animal-derived lysozyme may include, but is not limited to, egg white lysozyme, human or mammalian lysozyme; the plant source lysozyme can include, but is not limited to, lysozyme separated from plants such as pawpaw, turnip, fig, radish and the like; microbially-derived lysozyme may include, but is not limited to, beta-N-acetylhexosaminidase, amidase, lactamase, endopeptidase, beta-glucanase, mannanase, chitinase, and the like.
The lysozyme in the invention is preferably microbial lysozyme, and the inventor finds that the microbial lysozyme is selected to be the one which is easier to permeate into the product when acting on the product and is also less prone to generate flocculating substances on the surface of a membrane body, and the preferable lysozyme comprises but is not limited to beta-N-acetylhexosaminidase, beta-lactamase, beta-glucanase and chitinase.
The total mass concentration of the lysozyme is preferably between 0.01 and 0.5 percent. The volume concentration of specific lysozyme can be further preferred according to the minimum action concentration of different kinds of lysozyme, in an embodiment, for the validity of guaranteeing lysozyme, can choose 2 ~ 3 times of this lysozyme minimum action concentration usually, if: the N-acetylmuramidase is selected in the sterilization process, and can play a corresponding role as long as the total concentration of the N-acetylmuramidase is not less than 0.05%, and the N-acetylmuramidase is selected in the invention, and the total mass concentration of the N-acetylmuramidase is not less than 0.1% in the whole sterilization process. The minimum concentration of each lysozyme to be used can be obtained according to known materials or the prior art.
Furthermore, the antibiotic is one or more of beta-lactam antibiotics, macrolide antibiotics, aminoglycoside antibiotics, tetracycline antibiotics, sulfonamide antibiotics or nitrofuran antibiotics.
The beta-lactam antibiotics comprise one or more of but not limited to penicillins, cephalosporins, colistins, bacitracin zinc, vancomycin, virginiamycin and the like.
The penicillins of the present invention include, but are not limited to, penicillin sodium, penicillin potassium, ampicillin sodium, amoxicillin, piperacillin, penicillin V potassium, and the like.
The cephalosporins include but are not limited to cephalexin, cefadroxil, cefazolin sodium, cephradine, cefoperazone sodium, ceftazidime, ceftriaxone sodium and the like.
The macrolide antibiotics comprise one or more of erythromycin, acetylspiramycin, erythromycin ethylsuccinate, roxithromycin, midecamycin, azithromycin, clarithromycin, kitasamycin and the like.
The aminoglycoside antibiotics of the invention include but are not limited to one or more of gentamicin, amikacin, streptomycin, netilmicin, amikacin, tobramycin, neomycin, spectinomycin, micronomicin and the like.
The tetracycline antibiotics of the present invention include, but are not limited to, tetracycline, oxytetracycline, doxycycline, minocycline, and the like.
The sulfonamide antibiotics comprise one or more of, but not limited to, sulfathiazole, sulfamethazine, sulfamethoxypyridazine and the like.
The nitrofuran antibiotics of the invention include but are not limited to furazolidone.
In a preferred embodiment of the present invention, the antibiotic is preferably selected from the group consisting of penicillins and cephalosporins, wherein the penicillins include, but are not limited to, penicillin, benzathine, penicillin G, penicillin V, procaine, methicillin, dicloxacillin, flucloxacillin, cloxacillin, amoxicillin, ampicillin, amoxicillin, mezlocillin, azlocillin, piperacillin, carbenicillin, ticarcillin, nafcillin, imipenem (tyloxaden), meropenem, panipenem, and the like; cephalosporins include, but are not limited to, cefcyanomethyl, cephaloxyaminobenzyl, cephalobenzam, ceplaicin, cephalonine, ceftazidime, cephalothin, cefapirin, ceftriazine, cefotaxime, cefazedone, cefazolin, cephradine, cefixadine, ceftezole, cefaclor, cefonicid, cefprozil, cefuroxime, ceftizoxime, cefamandole, ceforanide, cefotiam, carbacephem: chlorocarbon, cefbuperazone, cefmetazole, cefminox, cefotetan, cefoxitin, cefcapene, cefixime, cefdinir, cefditoren, cefetamet, cefixime, cefepime, cefodizime, cefoperazone, cefotaxime, cefpodoxime, cefteram, ceftibuten, ceftiofur, cefotiarin, ceftizoxime, ceftriaxone, ceftazidime, cefpiramide, cefsulodin, moxalactam, Cefclidine, cefepime, cefotaxime, cefozopran, cefpirome, cefquinome, flomox, cefoperazine, ceftolom, ceftarom, cefazolin, cefcaper, cefcanel, ceftarol, cefpirome, ceftriaxone, cefrvir, cefsulpirtine, cefepime, cef78, ceftizoxime, ceftiofur □, ceftiofur pivoxil, ceftiofur 89mi, ceftiofur 898, ceftiofur, ceft.
The total mass of the antibiotic of the invention is preferably 0.01-0.7%. The specific preferable concentration can be further preferable according to the highest blood concentration of different antibiotics acting on human body, and is usually 1-2 times of the highest blood concentration.
The pH value of the sterilization liquid is preferably 4.5-6.0. The pH of the sterilizing solution can be adjusted by one or more agents commonly used in the field, such as citric acid, dilute sulfuric acid, dilute hydrochloric acid, acetic acid, lactic acid and fruit acid.
The reagent used for preparing the sterilizing liquid is water for injection.
The other purpose of the invention is to provide a sterilization method suitable for allogeneic and xenogeneic tissues and tissue engineering medical instruments, wherein allogeneic and xenogeneic tissues and tissue engineering medical instruments are soaked in the sterilization solution, and alkaline solution is slowly added to change the pH value of the sterilization solution to 7.5-8.5.
According to the invention, the lysozyme and the antibiotic are mixed to prepare the sterilizing solution, and under the condition of ensuring the effective sterilization concentration of the two components, the pH value of the sterilizing solution is changed in a titration mode, so that the permeability of the surface of the material is changed, the mixed sterilizing solution can fully act on the material, and the final sterilization effect is finally achieved.
The alkaline solution used in the method of the invention is preferably one or more of sodium hydroxide, calcium hydroxide, disodium hydrogen phosphate and sodium bicarbonate.
The slow addition method of the present invention may be any method conventional in the art, and in one embodiment, an alkaline reagent is used for the titration.
The titration time should be referenced to the effective action time of lysozyme and antibiotics and should ultimately be determined by microbial challenge tests.
After titration, the pH value of the soaking solution should stay between 7.5 and 8.5, and preferably, the soaking solution is fully mixed during titration, so that the concentration change is uniform.
In the titration process of the sterilization method, whether the lysozyme and the antibiotics in the sterilization solution are within the effective concentration or not needs to be ensured, namely the percentage content of the sterilization method can adjust the concentration of the titration solution and the sterilization solution at any time.
Furthermore, the soaking temperature is 25-38 ℃.
In order to further improve the final sterilization effect, the optimized initial microbial limit of the medical instruments of the allogeneic tissues and the xenogeneic tissues and the tissue engineering before sterilization is controlled within 100 cfu/g.
Further, the present invention provides a more specific method comprising the steps of:
(1) flushing the surfaces of the allogeneic tissues, the xenogeneic tissues and the tissue engineering medical instruments by using a buffering agent with the pH value of 5.6-7.8;
(2) completely soaking the allogeneic and xenogeneic tissues and tissue engineering medical instruments in a sterilization solution after washing, keeping the temperature constant at 25-38 ℃, and adjusting the pH value of the sterilization solution to 4.5-6.0;
(3) and slowly adding an alkaline solution to change the pH value of the sterilization solution to 7.5-8.5.
The buffer agent for washing can be selected from one or more of glycine-hydrochloric acid, phthalic acid-hydrochloric acid, disodium hydrogen phosphate-citric acid, citric acid-sodium hydroxide, acetic acid-sodium acetate, disodium hydrogen phosphate-potassium hydrogen phosphate and potassium dihydrogen phosphate-sodium hydroxide, and the tissue material cannot be pressed strongly or folded repeatedly in the washing process, so that the biological structure in the material is prevented from being damaged.
After the sterilization, the tissue material is taken out, the material can be cleaned by an ultrasonic cleaning method, and the cleaning solvent can be one of water for injection, normal saline, glycerol, dimethyl sulfoxide and cottonseed oil. According to the actual situation, the cleaning can be repeated for two to three times. The temperature of the solution should be controlled during the cleaning process to avoid damaging tissue materials due to overhigh temperature. And after the cleaning is finished, carrying out aseptic packaging according to the characteristics of different tissue materials and required preservation methods.
The advantages of the invention over the prior art are:
(1) the sterilization solution provided by the invention is prepared by combining lysozyme and antibiotics, the cell walls of microorganisms are damaged by the lysozyme, the materials are easy to permeate into the materials by virtue of the permeability of the antibiotics and the pH value of alkaline solution being slowly adjusted, the inhibition and destruction effects on the microorganisms are obvious, and the biological structure of the materials is not easy to destroy.
(2) The sterilization mode of the invention can effectively kill eyesMost of the previously known microorganisms, while preserving to the greatest extent the factors that contribute to the growth of the tissue in the tissue material, can meet the requirement that the sterilization level of the final product is less than 10-6The requirements of (1).
(3) The sterilization scope of the invention is wider, comprising: bacteria, fungi, chlamydia, mycoplasma, parasites, viruses, and the like.
(4) Traditional terminal sterilization (e.g., irradiation, ethylene oxide, dry heat, moist heat, ethanol, etc.) can severely damage the biological structure of the material. The method creatively changes the traditional sterilization of allogeneic and xenogeneic tissues and tissue engineering medical instruments for the first time. The sterilization process can maintain the matrix structure of the allogeneic and xenogeneic tissues and tissue engineering medical equipment and various growth factors in the tissue matrix completely, and the matrix structure and the growth factors are stored in large quantity to induce the implantation regeneration of autologous cell tissues in the recipient of the allogeneic and xenogeneic tissues and tissue engineering medical equipment implant, so as to form a natural scaffold structure, so that the tissues such as recipient capillary vessels and the like can spread and grow, and the survival of the allogeneic and xenogeneic tissues and tissue engineering medical equipment implant in the recipient can be promoted. Meanwhile, the medical device products of allogeneic and xenogeneic tissues and tissue engineering processed by the method are also good stem cell carriers, and can effectively provide good environment for differentiation and growth of stem cells. Through a large number of experiments, the TEMP product treated by the method has the characteristics of rapid cell tissue growth, rapid healing, low calcification, low infection, low absorption, micro scar after healing and the like.
Drawings
FIG. 1 shows the adherent growth of a basal lamina on the sterilized dermal matrix of example 1;
fig. 2 shows the adhesion growth of a basal lamina to the sterilized dermal matrix of comparative example 1.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
The following examples are given to ensure that the sterilization process is carried out in an absolutely sterile environment. The equipment involved in the whole sterilization process comprises: a biological safety cabinet, a precise constant-temperature water area device, a titrator, an ultrasonic cleaner, packaging equipment and the like. The whole operation process is completed based on aseptic operation (see YY/T0567.1-2005, part 1 of aseptic processing of medical products: general requirements).
Determination of residual sterilizing liquid
Because various lysozyme and antibiotics are used in the sterilization of the tissue engineering material, if the lysozyme and the antibiotics are remained in the material, the use of the product can be influenced to a certain extent. And whether the material is sterilized completely cannot be judged. The materials can be tested according to the method in the guidelines of the efficacy test for bacteriostatic agents in pharmacopoeia. The result should be that both bacteria and fungi can grow normally during the cultivation. If slow or no growth occurs, the experiment should be repeated. If the results are the same, the residual of the sterilizing liquid can be judged to be beyond the acceptable level. Adjustment of the ultrasonic cleaning time, the number of times of cleaning, and replacement of the cleaning solvent should be considered at this time.
Verification of Sterilization Effect
The tissue material can be inspected according to the sterility inspection method in pharmacopoeia, and the killing effect of the method on bacteria and fungi can be detected. For the killing effect of the virus, four indicators can be selected, namely the existence of lipid envelope and DNA or RNA genetic group. An indicator virus should be selected for some tolerance. Such as: pseudorabies virus (PRV), Sindbis virus, Porcine Parvovirus (PPV), encephalomyocarditis virus (EMCV), and the like.
Whether the allogenic and xenogenic materials have influence on the growth factors or the induction factors of the allogenic and xenogenic materials after the sterilization method is used can be proved by performing an ectopic induction forming experiment on an animal model.
Example 1
EXAMPLE 1 Sterilization of allogeneic dermal matrix
Step 1: preparing 500ml of mixed sterilized solution in a sterile environment, wherein the concentration of N-acetylmuramic polysaccharide hydrolase is 0.3mg/ml, the concentration of penicillin is 0.25mg/ml, and adjusting the pH value of the sterilized solution to 5.0 by using dilute hydrochloric acid;
step 2: taking out the dermal matrix to be sterilized (the thickness of the material should be less than 0.5mm, and the total weight should be less than 25g), flushing the surface of the material with normal saline, and flatly paving the material in a sterilizing solution;
and step 3: the temperature of the sterilized solution was kept constant at 35 ℃ and the sterilized solution was titrated by a titrator using a sodium hydroxide solution having a concentration of 50 mg/ml. The dropping time should be ensured to mix the solution evenly. The titration time is 2.5h, and the pH value of the final sterilizing solution is 7.5;
and 4, step 4: and (3) taking out the dermal matrix, cleaning the dermal matrix by using sterile normal saline, soaking the dermal matrix by using glycerol after cleaning, sealing and storing the dermal matrix, and simultaneously taking part of samples to test according to sterility test in pharmacopoeia to meet the requirements of the pharmacopoeia.
Comparative example 1 irradiation sterilized allogeneic dermal matrix
And (3) irradiating and sterilizing the dermal matrix to be sterilized (the thickness of the material is less than 0.5mm, and the total weight is less than 25g) by using 20-25KGy, taking part of samples, and checking the samples according to sterility test in pharmacopeia to meet the requirements of the pharmacopeia.
And (3) effect comparison:
the dermal matrices prepared by the two methods (example 1 and comparative example 1) were compared using the human basal lamina load test:
step 1: the dermal matrixes prepared by the two methods (example 1 and comparative example 1) are respectively taken and washed or rehydrated by using sterile physiological saline for standby;
step 2: taking about 1-3 g of healthy human subcutaneous basal layer skin, crushing to small particles with the physical diameter of 0.1-2.0 mm, and dyeing by using a biological dyeing agent; the pulverized skin pieces are divided into two parts and evenly smeared on two dermal matrixes;
and step 3: culturing in DMEM medium at 37 deg.C for 72 hr, taking out, and eluting with sterile physiological saline. Observing the growth condition of the small skin pieces;
the observation method comprises the following steps: after the dyed crushed skin pieces are uniformly coated, observing and counting by using an objective lens multiplied by 4 times, counting the number of the attached skin pieces in a visual field, randomly counting 5 observation points, and averagely counting 76 observation points. After the culture and elution, the results of fig. 1 and 2 show that the basal lamina skin sheets attached well to the sterilized dermal matrix of example 1 (fig. 1), but attached poorly to the irradiated and sterilized dermal matrix of comparative example 1, with a clear difference from example 1 (fig. 2). According to counting tests, the dermal matrix prepared in example 1 is attached to 65 spots on average, namely, the fusion differentiation rate of dermal sheet particles loaded by the dermal matrix prepared in example 1 reaches 85.5%; the dermal matrix prepared in comparative example 1 was attached to 11 sites on average, i.e., the dermal matrix loaded with the dermal patch small particles prepared in comparative example 1 was used with a fusion differentiation rate of only 14.5%.
Example 2 allogeneic tendon
1. Preparing 500ml of mixed sterilization liquid in a sterile environment, wherein the concentration of beta-lactamase is 0.5mg/ml, the concentration of penicillin is 0.25mg/ml, and adjusting the value of the sterilization liquid to 5.0 by using dilute hydrochloric acid;
2. taking out the variant tendon to be sterilized (the width of the material should be less than 2cm, and the thickness should be less than 0.3cm), washing the surface of the material with normal saline, and flatly paving the material in a sterilizing solution;
3. the temperature of the sterilized solution was kept constant at 35 ℃ and the sterilized solution was titrated by a titrator using a sodium hydroxide solution having a concentration of 50 mg/ml. The dropping time should be ensured to mix the solution evenly. The titration time is 2.5h, and the pH value of the final sterilizing solution is 7.5;
4. the allogeneic tendon is taken out and washed by sterile normal saline, is soaked by glycerol and is sealed for storage, and meanwhile, part of samples are taken to be checked according to sterility examination in pharmacopoeia, so that the requirements of the pharmacopoeia are met, and the clinical implantation effect is better.
Claims (10)
1. The sterilizing liquid suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering is characterized by comprising lysozyme with the mass concentration of 0.01-0.5% and antibiotics with the mass concentration of 0.01-0.7%.
2. The sterilization solution according to claim 1, wherein the pH of the sterilization solution is 4.5 to 6.0.
3. The sterilized solution of claim 1, wherein the medical devices of allogeneic and xenogeneic tissue and tissue engineering are:
(1) functional products that are not liquid-dispersible and/or soluble in water, lipids or/and alcohols;
(2) materials that do not contain moisture in the final product are excluded;
(3) does not include some tissue engineering materials with strong adsorption;
(4) does not comprise a matrix material needing the growth of the attached cells and a tissue engineering medical apparatus with drug loading implantation;
(5) excluding materials for functional bone graft, materials for cardiac pacemakers.
4. The sterilization liquid according to claim 1, wherein the lysozyme is one or more selected from the three classes of animal-derived lysozyme, plant-derived lysozyme and microorganism-derived lysozyme; preferably one or more of the following: egg white lysozyme, human or mammal lysozyme, lysozyme separated from plants such as papaya, turnip, fig, radish and the like, beta-N-acetylhexosaminidase, amidase, lactamase, endopeptidase, beta-glucanase, mannanase and chitinase; more preferably, the compound is selected from one or more of the following: beta-N-acetylhexosaminidase, beta-lactamase, beta-glucanase and chitinase.
5. The sterilization solution according to claim 1, wherein the antibiotic is one or more selected from the group consisting of β -lactam antibiotics, macrolide antibiotics, aminoglycoside antibiotics, tetracycline antibiotics, sulfonamide antibiotics, and nitrofuran antibiotics.
6. A sterilization method suitable for allogeneic and xenogeneic tissues and tissue engineering medical instruments is characterized in that the allogeneic and xenogeneic tissues and the tissue engineering medical instruments are soaked in the sterilization solution of claim 1, and alkaline solution is slowly added to change the pH value of the sterilization solution to 7.5-8.5.
7. A sterilization process according to claim 6, wherein the temperature of the soaking is 25 to 38 ℃.
8. Sterilisation method according to claim 6, characterised in that it comprises the steps of:
(1) flushing the surfaces of the allogeneic tissues, the xenogeneic tissues and the tissue engineering medical instruments by using a buffering agent with the pH value of 5.6-7.8;
(2) completely soaking the allogeneic and xenogeneic tissues and tissue engineering medical instruments in the sterile liquid of claim 1 after washing, keeping the temperature constant at 25-38 ℃, and adjusting the pH value of the sterile liquid to 4.5-6.0;
(3) and slowly adding an alkaline solution to change the pH value of the sterilization solution to 7.5-8.5.
9. A sterilization method according to any one of claims 6 to 8, wherein the alkaline solution is one or more of sodium hydroxide, calcium hydroxide, disodium hydrogen phosphate and sodium bicarbonate.
10. The sterilization method according to claim 8, wherein the buffer is one or more selected from glycine-hydrochloric acid, phthalic acid-hydrochloric acid, disodium hydrogen phosphate-citric acid, citric acid-sodium hydroxide, acetic acid-sodium acetate, disodium hydrogen phosphate-potassium hydrogen phosphate, and potassium dihydrogen phosphate-sodium hydroxide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910371621.3A CN110692647A (en) | 2019-05-06 | 2019-05-06 | Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910371621.3A CN110692647A (en) | 2019-05-06 | 2019-05-06 | Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110692647A true CN110692647A (en) | 2020-01-17 |
Family
ID=69193043
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910371621.3A Pending CN110692647A (en) | 2019-05-06 | 2019-05-06 | Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110692647A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991001723A1 (en) * | 1989-08-09 | 1991-02-21 | Osteotech, Inc. | Process for disinfecting musculoskeletal tissue and tissues prepared thereby |
| CN1557485A (en) * | 2004-01-30 | 2004-12-29 | 大连帝恩生物工程有限公司 | Application of gene recombined human lysozyme in eliminating pathogenic microorganism infection |
| CN1785428A (en) * | 2004-12-08 | 2006-06-14 | 广州威尔曼新药开发中心有限公司 | New lysozyme compound injection preparation |
| CN101389358A (en) * | 2006-01-10 | 2009-03-18 | 爱罗索斯公司 | Apparatus and methods for treating allograft products |
| CN100482216C (en) * | 1999-12-16 | 2009-04-29 | 德马全兹股份有限公司 | Use of hydroxide-releasing agents as skin permeation enhancers |
| CN108925616A (en) * | 2018-08-03 | 2018-12-04 | 嘉应学院 | A kind of lysozyme composite preservative and the preparation method and application thereof |
-
2019
- 2019-05-06 CN CN201910371621.3A patent/CN110692647A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991001723A1 (en) * | 1989-08-09 | 1991-02-21 | Osteotech, Inc. | Process for disinfecting musculoskeletal tissue and tissues prepared thereby |
| CN100482216C (en) * | 1999-12-16 | 2009-04-29 | 德马全兹股份有限公司 | Use of hydroxide-releasing agents as skin permeation enhancers |
| CN1557485A (en) * | 2004-01-30 | 2004-12-29 | 大连帝恩生物工程有限公司 | Application of gene recombined human lysozyme in eliminating pathogenic microorganism infection |
| CN1785428A (en) * | 2004-12-08 | 2006-06-14 | 广州威尔曼新药开发中心有限公司 | New lysozyme compound injection preparation |
| CN101389358A (en) * | 2006-01-10 | 2009-03-18 | 爱罗索斯公司 | Apparatus and methods for treating allograft products |
| CN108925616A (en) * | 2018-08-03 | 2018-12-04 | 嘉应学院 | A kind of lysozyme composite preservative and the preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2019202651B2 (en) | Processed adipose tissue | |
| US20220160937A1 (en) | Rapid allograft treatment systems and methods | |
| US5139527A (en) | Biologic absorbable implant material for filling and closing soft tissue cavities and method of its preparation | |
| US5788941A (en) | Method of sterilization of bone tussue | |
| US8574825B2 (en) | Process for demineralization of bone matrix with preservation of natural growth factors | |
| JP6606429B2 (en) | Decellularized biomaterial from non-mammalian tissue | |
| US10758642B2 (en) | Sterilization process | |
| US20150010510A1 (en) | Bone-Derived Extra Cellular Matrix Gel | |
| EP3597200A1 (en) | Bioresorbable biological matrix for repairing bone tissue defects and method for the production thereof | |
| CA2263557C (en) | Implant consisting of a carrier material containing medical substances and method to produce such an implant | |
| Das et al. | Tissue-derived decellularized extracellular matrices toward cartilage repair and regeneration | |
| RU2526429C1 (en) | Method of manufacturing bone implants | |
| CA1333050C (en) | Biologic absorbable implant material for filling and closing soft-tissue cavities and method of its preparation | |
| CN104474573A (en) | Biological mask and preparation method thereof | |
| CN110692647A (en) | Sterilization liquid and sterilization method suitable for medical instruments of allogeneic and xenogeneic tissues and tissue engineering | |
| RU2691983C1 (en) | Method for purification, modification and sterilization of bone tissue and skin matrix derivatives using supercritical fluid | |
| US20180280575A1 (en) | Rapid acoustic tissue processing methods, systems, and devices | |
| CA2963554C (en) | Polymer solution for visco-supplementation | |
| EP3188596B1 (en) | Human dermis, preparation and use thereof | |
| CN109602932B (en) | Method for disinfecting decellularized protein scaffold | |
| RU2774579C1 (en) | Method for sterilizing acellular matrixes | |
| RU2712701C1 (en) | Implant for bone tissue replacement | |
| YUAN et al. | Vancomycin Loaded Alginate/Cockle Shell Powder Nanobiocomposite Bone Scaffold for Antibacterial and Drug Release Evaluation | |
| Mardana et al. | Chronic osteomyelitis treatment with PerOssal®: a literature review | |
| CA3217558A1 (en) | Sterilized multicomponent composition for removal of particles |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200117 |
|
| RJ01 | Rejection of invention patent application after publication |