CN115006596A - Process for treating biological tissue sheet and biological tissue sheet obtained by the treatment process - Google Patents
Process for treating biological tissue sheet and biological tissue sheet obtained by the treatment process Download PDFInfo
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
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Abstract
A treatment process of a biological tissue sheet and the biological tissue sheet obtained by the treatment process relate to the technical field of biological material treatment. The treatment process of the biological tissue sheet comprises the following steps: providing a biological tissue sheet soaked in a glutaraldehyde solution; the biological tissue sheet soaked in the glutaraldehyde solution is firstly fixed by glutaraldehyde crosslinking; and taking the biological tissue sheet out of the glutaraldehyde solution, then cleaning the biological tissue sheet by adopting a NaCl aqueous solution, and mixing and reacting the biological tissue sheet with a glycine solution after cleaning. The treatment process can lead the biological tissue sheet to have less residual glutaraldehyde and lower cytotoxicity.
Description
Technical Field
The application relates to the technical field of biological material treatment, in particular to a treatment process of a biological tissue sheet and the biological tissue sheet obtained by the treatment process.
Background
The animal-derived medical instrument refers to a medical instrument which is made of an inanimate animal-derived biological material as a main material, and researches show that the animal-derived medical instrument has better biocompatibility and bioactivity compared with the medical instrument made of a non-animal-derived material (such as metal, a high polymer material or an inorganic material).
The biological tissue sheet can be generally used for biological patches and heart valves, and the biological patches can be applied to neurosurgery, endocranial repair, pleural repair, skull repair, large-area burn, breast repair and the like. In order to improve the mechanical properties of the biological tissue sheet, the biological tissue sheet generally needs to be crosslinked by glutaraldehyde. However, glutaraldehyde has strong cytotoxicity, and since the biological tissue sheet generally needs to be implanted into a human body for use, the residual glutaraldehyde easily causes certain safety risks.
Disclosure of Invention
The application provides a biological tissue sheet treatment process and a biological tissue sheet obtained by the treatment process, wherein the treatment process can ensure that the biological tissue sheet has less residual glutaraldehyde and has lower cytotoxicity.
In a first aspect, the present application provides a process for treating a biological tissue sheet, comprising the steps of:
providing a biological tissue sheet soaked in a glutaraldehyde solution; the biological tissue sheet soaked in the glutaraldehyde solution is firstly fixed by glutaraldehyde crosslinking;
and taking the biological tissue sheet out of the glutaraldehyde solution, then cleaning the biological tissue sheet by adopting a NaCl aqueous solution, and mixing and reacting the biological tissue sheet with a glycine solution after cleaning.
In some possible embodiments, the step of cross-linking and fixing the biological tissue sheet with glutaraldehyde comprises:
reacting the biological tissue sheet with a glutaraldehyde solution with the mass concentration of 0.5-0.7% for a first preset time, and then reacting with a glutaraldehyde solution with the mass concentration of 0.2-0.4% for a second preset time, wherein the second preset time is longer than the first preset time; optionally, the first preset time is 40-50 hours, and the second preset time is 320-360 hours.
In some possible embodiments, the biological tissue sheet after the cross-linking treatment is soaked in a glutaraldehyde solution with the mass concentration of 0.5-0.7%.
In some possible embodiments, the mass concentration of the glycine solution is 0.05-5%, and the mixing reaction time of the biological tissue sheet and the glycine solution is more than or equal to 1 h; optionally, the mixing reaction time of the biological tissue sheet and the glycine solution is 1-6 h.
In some possible embodiments, the mass concentration of the glycine solution is 0.05-0.5%, and the mixing reaction time of the biological tissue sheet and the glycine solution is 3-6 h.
In some possible embodiments, the mass concentration of the NaCl aqueous solution is 6-12 g/L.
In some possible embodiments, the aqueous NaCl solution has a mass concentration of 9 g/L.
In some possible embodiments, the number of times the biological tissue sheet is washed with the aqueous NaCl solution is 1-8 times.
In some possible embodiments, the biological tissue sheet comprises at least one of pericardium, dermal matrix, mesentery, and aortic valve.
In a second aspect, the present application provides a biological tissue sheet obtained by the treatment process for a biological tissue sheet of the first aspect.
The application has at least the following beneficial effects:
according to the treatment process of the biological tissue sheet, the biological tissue sheet soaked in the glutaraldehyde solution is fixed by glutaraldehyde crosslinking, and the biological tissue sheet fixed by glutaraldehyde crosslinking has good mechanical properties. The biological tissue sheet after being cross-linked and fixed by the glutaraldehyde is soaked in a glutaraldehyde solution, so that the biological tissue sheet can play the roles of disinfection, sterilization and corrosion prevention, and is convenient for subsequent use. After the biological tissue sheet is taken out from the glutaraldehyde solution, the NaCl aqueous solution is firstly adopted for cleaning, so that most glutaraldehyde residual solution can be cleaned, and the NaCl aqueous solution remained in the biological tissue sheet is not easy to influence cells. After the biological tissue sheet is cleaned by NaCl aqueous solution, the biological tissue sheet is mixed with glycine solution for reaction, and the amino group of the glycine can react with the aldehyde group of the residual glutaraldehyde, so that the residual glutaraldehyde in the biological tissue sheet can be reduced, and the biological tissue sheet has lower cytotoxicity.
Drawings
It is to be understood that the following drawings illustrate only certain embodiments of this invention and are therefore not to be considered limiting of its scope, for those skilled in the art to which it pertains without inventive faculty, and that other related drawings may be derived therefrom.
FIG. 1 is a chromatogram corresponding to example 1 of the present application;
FIG. 2 is a chromatogram corresponding to example 2 of the present application;
FIG. 3 is a chromatogram corresponding to example 3 of the present application;
FIG. 4 is a chromatogram corresponding to example 4 of the present application;
FIG. 5 is a chromatogram corresponding to example 5 of the present application;
FIG. 6 is a chromatogram corresponding to example 6 of the present application;
FIG. 7 is a chromatogram corresponding to example 7 of the present application;
FIG. 8 is a chromatogram corresponding to example 8 of the present application;
FIG. 9 is a chromatogram corresponding to example 9 of the present application;
FIG. 10 is a chromatogram corresponding to example 10 of the present application;
FIG. 11 is a chromatogram corresponding to example 11 of the present application;
FIG. 12 is a chromatogram corresponding to example 12 of the present application;
FIG. 13 is a chromatogram corresponding to comparative example 1 of the present application.
Detailed Description
Embodiments of the present application will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present application and should not be construed as limiting the scope of the present application. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The following will specifically describe a process for treating a biological tissue sheet according to an embodiment of the present application and a biological tissue sheet obtained by the process:
in a first aspect, an embodiment of the present application provides a process for processing a biological tissue sheet, including the following steps:
(1) providing a biological tissue sheet soaked in a glutaraldehyde solution; the biological tissue sheet soaked in the glutaraldehyde solution is firstly fixed by glutaraldehyde crosslinking.
The biological tissue sheet soaked in the glutaraldehyde solution is fixed by glutaraldehyde crosslinking, and the biological tissue sheet fixed by glutaraldehyde crosslinking has better mechanical property. The biological tissue sheet after being cross-linked and fixed by the glutaraldehyde is soaked in a glutaraldehyde solution, so that the biological tissue sheet can play the roles of disinfection, sterilization and corrosion prevention, and is convenient for subsequent use. Wherein the biological tissue sheet comprises at least one of a pericardium, a dermal matrix, a mesentery, and an aortic valve.
Optionally, the biological tissue sheet is soaked in a glutaraldehyde solution with a mass concentration of 0.5-0.7%, for example, at any one of or a value between 0.5%, 0.55%, 0.6%, 0.625%, 0.65%, and 0.7%.
In some embodiments, the step of cross-linking and fixing the biological tissue sheet with glutaraldehyde comprises: reacting the biological tissue sheet with a glutaraldehyde solution with the mass concentration of 0.5-0.7% for a first preset time, and then reacting with a glutaraldehyde solution with the mass concentration of 0.2-0.4% for a second preset time, wherein the second preset time is longer than the first preset time.
For convenience of explanation, the first reaction is defined as the reaction of the biological tissue sheet with a glutaraldehyde solution having a mass concentration of 0.5 to 0.7%, and the second reaction is defined as the reaction of the biological tissue sheet with a glutaraldehyde solution having a mass concentration of 0.2 to 0.4%.
The biological tissue sheet is firstly reacted with glutaraldehyde solution with the mass concentration of 0.5-0.7% for a first preset time, so that the biological tissue sheet can reach a certain crosslinking degree. And then, the biological tissue sheet is reacted with a glutaraldehyde solution with the concentration of 0.2-0.4% for a second preset time, the concentration of glutaraldehyde during the second reaction is reduced, the reaction time is prolonged, and the biological tissue sheet and glutaraldehyde can be fully reacted to achieve a better and appropriate degree of crosslinking. If the concentration of glutaraldehyde in the second reaction is too low, the degree of crosslinking of the biological tissue sheet may be insufficient.
Illustratively, the mass concentration of glutaraldehyde of the first reaction is any one of, or a value between, 0.5%, 0.55%, 0.6%, 0.65%, and 0.7%.
Illustratively, the mass concentration of glutaraldehyde of the second reaction is any one of, or a value between, 0.2%, 0.25%, 0.3%, 0.35%, and 0.4%.
In addition, it should be noted that if the second predetermined time is too long, the production efficiency may be lowered by lengthening the reaction time since the biological tissue sheet has already reached a good degree of crosslinking. Optionally, the first preset time is 40-50 hours, and the second preset time is 320-360 hours.
For example, the first preset time is 40h, 42h, 44h, 46h, 48h or 50 h; the second preset time is 320h, 324h, 328h, 332h, 336h, 340h, 344h, 348h, 352h, 356h or 360 h.
(2) And taking the biological tissue sheet out of the glutaraldehyde solution, then cleaning the biological tissue sheet by adopting a NaCl aqueous solution, and mixing and reacting the biological tissue sheet with a glycine solution after cleaning.
Wherein, after the biological tissue sheet is crosslinked and fixed by glutaraldehyde, the biological tissue sheet generally can not directly enter the next procedure for treatment. Therefore, the biological tissue sheet which is fixed by glutaraldehyde crosslinking is firstly soaked in a glutaraldehyde solution, and when the next procedure is carried out, the biological tissue sheet is taken out of the glutaraldehyde solution, then is cleaned by a NaCl aqueous solution, and is mixed with a glycine solution for reaction after being cleaned, so that the residual amount of glutaraldehyde in the biological tissue sheet is less, and the biological tissue sheet has lower cytotoxicity.
After the biological tissue sheet is taken out from the glutaraldehyde solution, the NaCl aqueous solution is firstly adopted for cleaning, so that most glutaraldehyde residual solution can be cleaned, and the NaCl aqueous solution remained in the biological tissue sheet is not easy to influence cells. After the biological tissue sheet is cleaned by NaCl aqueous solution, the biological tissue sheet is mixed with glycine solution for reaction, and the amino group of the glycine can react with the aldehyde group of the residual glutaraldehyde, so that the residual glutaraldehyde in the biological tissue sheet is less, and the biological tissue sheet has lower cytotoxicity.
The intracellular fluid in the human body has a certain concentration, blood cells can be ruptured in pure water, and the blood cells can lose water in a high-concentration NaCl aqueous solution, so that the mass concentration of the NaCl aqueous solution is 6-12 g/L optionally, the NaCl aqueous solution with the concentration is close to the osmotic pressure of cells of the human body, and the NaCl aqueous solution in the concentration range is adopted to clean the biological tissue sheet, so that the cells are not easily affected by the residual NaCl aqueous solution on the biological tissue sheet implanted into the human body even if the NaCl aqueous solution is residual on the biological tissue sheet.
Illustratively, the aqueous NaCl solution has a mass concentration of 6g/L, 7g/L, 8g/L, 9g/L, 10g/L, 11g/L, or 12 g/L. In one embodiment, the aqueous NaCl solution has a mass concentration of 9 g/L. The NaCl aqueous solution with the mass concentration of 9g/L is the physiological saline, and the NaCl aqueous solution does not influence cells.
Optionally, the biological tissue sheet is washed with the NaCl aqueous solution 1-8 times. In the cleaning process, the residual glutaraldehyde on the biological tissue sheet can be better cleaned by more times of cleaning. For example, the number of times the biological tissue sheet is washed with the NaCl aqueous solution is 1, 2, 3, 4, 5, 6, 7 or 8 times.
Further, the inventors of the present application have found in their studies that the concentration of glycine solution and the length of time the biological tissue sheet is reacted with glycine solution affect the residual glutaraldehyde content and cytotoxicity of the final biological tissue sheet. In some possible embodiments, the glycine solution has a mass concentration of 0.05 to 5%, such as any one or a value between any two of 0.05, 0.1%, 0.15%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, and 5%.
In the research of the inventor, the inventor discovers that after the biological tissue sheet is reacted with a glutaraldehyde solution with the mass concentration of 0.5-0.7% for 40-50 hours, the biological tissue sheet is reacted with a glutaraldehyde solution with the mass concentration of 0.2-0.4% for 320-360 hours, then the biological tissue sheet is soaked in the glutaraldehyde solution with the mass concentration of 0.5-0.7%, and then the biological tissue sheet is washed by a NaCl aqueous solution, if the concentration of the glycine solution reacted with the biological tissue sheet is too small, the cytotoxicity of the biological tissue sheet is reduced to a small extent; if the concentration of glycine reacted with the biological tissue sheet is too high, the pH of the entire system may be changed, and although reduction of the cytotoxicity of the biological tissue sheet may also be achieved, the effect is not very good.
Furthermore, the time for mixing and reacting the biological tissue sheet and the glycine solution is 1-6 h, such as 1h, 2h, 3h, 4h, 5h or 6 h. In the research of the inventor, when the time for mixing and reacting the biological tissue sheet with the glycine solution is too long, for example, 24 hours, the cytotoxicity of the biological tissue sheet is not greatly reduced from 6 hours, and the too long time affects the production efficiency. Therefore, the time of the mixed reaction of the biological tissue sheet and the glycine is 1-6 h, the effect of better reducing the cytotoxicity of the biological tissue sheet can be achieved, and the influence on the production efficiency due to overlong time can be avoided.
In some embodiments, the mass concentration of the glycine solution is 0.05-0.5%, and the mixing reaction time of the biological tissue sheet and the glycine solution is 3-6 h. Under these conditions, the residual amount of glutaraldehyde is relatively small.
In a second aspect, the present application provides a biological tissue sheet obtained by the treatment process for a biological tissue sheet of the first aspect.
The biological tissue sheet obtained by the treatment process of the biological tissue sheet of the first aspect has not only better mechanical properties, but also lower cytotoxicity.
The processing of the biological tissue sheet of the present application and the biological tissue sheet obtained by the processing are further described in detail below with reference to examples.
Example 1
The embodiment provides a treatment process of a biological tissue sheet, which comprises the following steps:
providing a biological tissue sheet soaked in a glutaraldehyde solution with the mass concentration of 0.6%; the biological tissue sheet soaked in the glutaraldehyde solution is obtained by firstly reacting with 0.6% glutaraldehyde solution for 48 hours and then reacting with 0.25% glutaraldehyde for 336 hours.
And taking the biological tissue sheet soaked in the glutaraldehyde solution out of the glutaraldehyde solution, then washing the biological tissue sheet for 6 times by using a NaCl aqueous solution with the mass concentration of 9g/L, and mixing and reacting the biological tissue sheet with a glycine solution with the mass concentration of 0.1% for 1 hour after washing.
Example 2
Example 2 provides a process for treating a biological tissue sheet, which is different from example 1 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours respectively.
Example 3
Example 5 provides a process for treating a biological tissue sheet, which differs from example 1 only in that the concentration of glycine solution in example 5 is 0.15%.
Example 4
Example 4 provides a process for treating a biological tissue sheet, which is different from example 3 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours respectively.
Example 5
Example 5 provides a process for treating a biological tissue sheet, which differs from example 1 only in that the concentration of glycine solution in example 5 is 0.05%.
Example 6
Example 6 provides a process for treating a biological tissue sheet, which is different from example 5 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours respectively.
Example 7
Example 7 provides a process for treating a biological tissue sheet, which is different from example 1 only in that the concentration of glycine solution in example 7 is 0.5%.
Example 8
Example 8 provides a process for treating a biological tissue sheet, which is different from example 7 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours.
Example 9
Example 9 provides a process for treating a biological tissue sheet, which is different from example 1 only in that the concentration of the glycine solution in example 9 is 1%.
Example 10
Example 10 provides a process for treating a biological tissue sheet, which is different from example 9 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours.
Example 11
Example 11 provides a process for treating a biological tissue sheet, which is different from example 1 only in that the concentration of the glycine solution in example 11 is 5%.
Example 12
Example 12 provides a process for treating a biological tissue sheet, which is different from example 11 only in that the time for mixing and reacting the biological tissue sheet with the glycine solution is 6 hours.
Example 13
The embodiment provides a treatment process of a biological tissue sheet, which comprises the following steps:
providing a biological tissue sheet soaked in a glutaraldehyde solution with the mass concentration of 0.7%; the biological tissue sheet soaked in the glutaraldehyde solution is obtained by firstly reacting with 0.5% glutaraldehyde solution for 40h and then reacting with 0.2% glutaraldehyde for 360 h.
And taking the biological tissue sheet soaked in the glutaraldehyde solution out of the glutaraldehyde solution, then washing the biological tissue sheet for 3 times by using a NaCl aqueous solution with the mass concentration of 12g/L, and mixing and reacting the biological tissue sheet with a glycine solution with the mass concentration of 0.1% for 1 hour after washing.
Example 14
The embodiment provides a treatment process of a biological tissue sheet, which comprises the following steps:
providing a biological tissue sheet soaked in a glutaraldehyde solution with the mass concentration of 0.5%; the biological tissue sheet soaked in the glutaraldehyde solution is obtained by firstly reacting with 0.7% glutaraldehyde solution for 45 hours and then reacting with 0.4% glutaraldehyde for 320 hours.
And taking the biological tissue sheet soaked in the glutaraldehyde solution out of the glutaraldehyde solution, then washing the biological tissue sheet for 5 times by using a NaCl aqueous solution with the mass concentration of 10g/L, and mixing and reacting the biological tissue sheet with a glycine solution with the mass concentration of 0.1% for 1 hour after washing.
Comparative example 1
This comparative example uses the biological tissue sheet of example 1 immersed in a glutaraldehyde solution at a mass concentration of 0.6% as a control.
Test examples
(1) The biological tissue sheets obtained by the treatment process of the biological tissue sheets of examples 1 to 12 and the control biological tissue sheet were cut into 5mm × 25mm pieces with sterile ophthalmic scissors and washed with physiological saline 3 times for 1 minute each time in a super clean bench. After the completion of the washing, the sheet was cut into small pieces of biological tissue at a length of 6cm 2 MEM medium containing fetal bovine serum was added at a ratio of/ml, and the mixture was extracted by shaking at a constant temperature of 37. + -. 1 ℃ for 72. + -.2 hours at a shaking speed of 60 rpm. And after leaching, separating the sample from the liquid, cooling to room temperature, and taking the liquid as a leaching solution. In vitro cytotoxicity was also tested with reference to GB/T16886.5-2017 (section 5 of the evaluation of medical devices in biology: in vitro cytotoxicity test) leach liquor, and the cytotoxicity was tested by the CCK-8 method, the results of which are reported in Table 1. Wherein, the cell survival rate obtained by the test is more than or equal to 70 percent and qualified, and the cell survival rate obtained by the test is less than 70 percent and unqualified.
(2) The leaching liquor was prepared in the same manner as in test example (1), and the residual amount of glutaraldehyde was measured in the leaching liquor, wherein the residual amount of glutaraldehyde was measured by 3204 glutaraldehyde residue measurement in accordance with the general rule of "chinese pharmacopoeia" 2020 edition, and the test results are shown in table 1. The liquid chromatograms of examples 1 to 12 and comparative example 1 are shown in fig. 1 to 13, respectively. The difference between the peak discharge times in fig. 1 to 4 and 11 and the peak discharge times in fig. 5 to 10 and 12 and 13 is due to the difference between the column size and the packing size, and the column size used in fig. 1 to 4 and 11 is 4.6 μm (inner diameter) × 150mm (length), and the packing particle size is 4 μm; the column used in FIGS. 5 to 10, 12 and 13 had a size of 4.6. mu. m.times.250 mm, and the filler had a particle size of 5 μm.
TABLE 1 test results
From the results in table 1, it can be seen that the treatment process for the biological tissue sheet according to the embodiment of the present application can effectively reduce glutaraldehyde residue, and make the biological tissue sheet have low cytotoxicity, which meets the relevant requirements.
The foregoing is merely exemplary of the present application and is not intended to limit the present application, which may be modified or varied by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A process for treating a biological tissue sheet, comprising the steps of:
providing a biological tissue sheet soaked in a glutaraldehyde solution; the biological tissue sheet soaked in the glutaraldehyde solution is fixed by glutaraldehyde crosslinking;
and taking out the biological tissue sheet from the glutaraldehyde solution, then cleaning the biological tissue sheet by adopting a NaCl aqueous solution, and mixing and reacting the biological tissue sheet with a glycine solution after cleaning.
2. The process for treating a biological tissue sheet according to claim 1, wherein the step of cross-linking and fixing the biological tissue sheet with glutaraldehyde comprises:
reacting the biological tissue sheet with a glutaraldehyde solution with the mass concentration of 0.5-0.7% for a first preset time, and then reacting with a glutaraldehyde solution with the mass concentration of 0.2-0.4% for a second preset time, wherein the second preset time is longer than the first preset time; optionally, the first preset time is 40-50 hours, and the second preset time is 320-360 hours.
3. The process for treating a biological tissue sheet according to claim 1, wherein the biological tissue sheet is immersed in a glutaraldehyde solution having a mass concentration of 0.5 to 0.7%.
4. The treatment process of the biological tissue sheet according to any one of claims 1 to 3, wherein the mass concentration of the glycine solution is 0.05 to 5%, and the mixing reaction time of the biological tissue sheet and the glycine solution is not less than 1 h; optionally, the mixing reaction time of the biological tissue sheet and the glycine solution is 1-6 h.
5. The process for treating the biological tissue sheet according to claim 4, wherein the mass concentration of the glycine solution is 0.05-0.5%, and the mixing reaction time of the biological tissue sheet and the glycine solution is 3-6 h.
6. The process for treating a biological tissue sheet according to any one of claims 1 to 3, wherein the aqueous NaCl solution has a mass concentration of 6 to 12 g/L.
7. The process for treating a biological tissue sheet according to claim 6, wherein the aqueous NaCl solution has a mass concentration of 9 g/L.
8. The process for treating a biological tissue sheet according to any one of claims 1 to 3, wherein the biological tissue sheet is washed with the NaCl aqueous solution 1 to 8 times.
9. The process for treating a biological tissue sheet according to any one of claims 1 to 3, wherein the biological tissue sheet comprises at least one of pericardium, dermal matrix, mesentery, and aortic valve.
10. A biological tissue sheet obtained by the treatment process for a biological tissue sheet according to any one of claims 1 to 9.
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| CN117045865A (en) * | 2023-09-25 | 2023-11-14 | 重庆生物智能制造研究院 | Method for removing residual glutaraldehyde in glutaraldehyde-crosslinked biological material |
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