CN116983459A - Preparation method of low-immunogenicity absorbable suture based on tendon of castoreum - Google Patents
Preparation method of low-immunogenicity absorbable suture based on tendon of castoreum Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L17/00—Materials for surgical sutures or for ligaturing blood vessels ; Materials for prostheses or catheters
- A61L17/06—At least partially resorbable materials
- A61L17/08—At least partially resorbable materials of animal origin, e.g. catgut, collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L17/00—Materials for surgical sutures or for ligaturing blood vessels ; Materials for prostheses or catheters
- A61L17/14—Post-treatment to improve physical properties
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- Life Sciences & Earth Sciences (AREA)
- Surgery (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
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Abstract
The invention relates to the technical field of absorbable sutures, in particular to a preparation method of a low-immunogenicity absorbable suture based on a tendon of a beaver. In particular, the present invention relates to treating isolated beaver tendons with an acid enzyme method to selectively hydrolyze telopeptide of collagen, thereby reducing immunogenicity of suture based on the beaver tendons, and simultaneously treating the isolated beaver tendons with glutaraldehyde as a cross-linking agent at a specific concentration to increase strength of the tendons, thereby improving tensile strength of the suture. The invention realizes the low-immunogenicity absorbable suture line based on the beaver tendon with optimal comprehensive performance of immunogenicity and mechanical strength by determining the proper treatment time for removing partial telopeptide of the tendon material under the specific condition by using pepsin solution under the specific feed-liquid ratio and determining the proper concentration of glutaraldehyde as a cross-linking agent for enhancing the mechanical performance of the tendon material under the condition of removing partial telopeptide.
Description
Technical Field
The invention relates to the technical field of absorbable sutures, in particular to a preparation method of a low-immunogenicity absorbable suture based on a tendon of a beaver.
Background
Medical suture is a common linear material and is widely applied to various surgical operations for suturing wounds and connecting tissues. With the continuous progress of science and technology, the suture material is currently subjected to four generations of development processes; the first generation is silk thread, the second generation is catgut thread, the third generation is chemical synthesis absorbable suture (PGA, PGLA, PLA), and the fourth generation is collagen absorbable suture. The medical suture lines after the second generation are absorbable suture lines, wherein the catgut lines have different degrees of rejection phenomena on human bodies, are easy to cause postoperative rejection reaction of patients, have low tensile strength, and clinically show various postoperative adverse reactions. Chemical synthetic sutures have been widely used clinically because of their good degradability and absorbability, and have achieved good results. However, chemical residues with different degrees still exist after the chemical synthesis suture is implanted, subcutaneous induration of the organism is easy to cause, and the problems of subcutaneous itching and the like exist after the operation of part of patients.
Castors belong to mammals and have been reported to be 98% similar in gene to humans. It was observed under a microscope that tendons consisted of two parts, namely tendon cells and collagen fibers. The tail tendon of the beaver consists of 95-96% of collagen, belongs to type I collagen, is clearly researched and widely used for biomedical materials. Collagen is a family of biological ultra-large molecular proteins that are widely found in the extracellular matrix, with a typical triple helix structure. Different types of collagen have great complexity and diversity in terms of structure, non-helical domain, assembly, function, etc. The unique structure and chemical composition of collagen imparts a number of unique properties and functions. These properties and functions make it widely used in various fields such as biomedical materials, drug delivery vehicles, tissue engineering, cosmetics, foods, and the like. Under the conditions of physiological state such as pH value and temperature, the collagen is not easy to degrade by general proteolytic enzyme, but under the physiological state, collagenase can degrade collagen in extracellular matrix, so that three-strand helix of the collagen is split into two fragments; at 37 ℃, the two fragments are easily broken down into small peptides and amino acids by collagenases and other proteases. Thus, it is clear from the nature of collagen that tendons are absorbable materials that can be used as raw materials for surgical sutures. The special three-dimensional structure of the collagen makes the collagen of the adult animal extremely stable. Collagen is insoluble in water, dilute acid, dilute alkali and alcohol, which provides a very advantageous condition for tendon washing, degreasing and disinfection. The research on the property of the tendon has the encouraging development prospect of applying the tail tendon of the beaver to the preparation of surgical suture.
Chinese patent No. CN95103686.6 (publication No. CN1113159 a) discloses for the first time a purely natural absorbable collagen suture, which is prepared by killing animals, sterilizing to extract spinal striated muscle tendon, repeatedly washing the extracted tendon tissue with sterile water until it is silvery white; the cleaned tendon tissue is then classified into fibers of the desired thickness and length by manual or mechanical means, and is soaked in 75% ethanol for 24 hours to fix, dehydrate, defat and sterilize.
The Chinese patent No. CN201510393691.0 (publication No. CN 105148318A) discloses a preparation method of medical absorbable collagen suture of beaver, which comprises the following steps: 1) Stripping off tendons; 2) Rinsing tendons with purified water; 3) Soaking in ethanol, dehydrating, and fixing; 4) Treating with NaOH solution, and then cleaning with purified water; 5) The mixture was dehydrated and fixed by soaking in 75% ethanol, and the mixture was stored in 75% ethanol solution.
However, the sutures prepared by these methods still have a weak immunogenicity inherent to collagen, and the tensile strength of the suture is not high enough.
Disclosure of Invention
In order to solve the problem of immunogenicity of the suture based on the tendon of the beaver, the present invention treats the isolated tendon of the beaver with an acid enzyme method to selectively hydrolyze the telopeptide of collagen, which is known in the art as the greatest cause of the immune response by collagen, thereby reducing the immunogenicity of the suture based on the tendon of the beaver.
In order to solve the problem of tensile strength of the suture based on the tendon of the beaver, the invention treats the separated tendon of the beaver by using glutaraldehyde with specific concentration as a cross-linking agent so as to increase the strength of the tendon, thereby improving the tensile strength of the suture.
In particular, the invention provides a preparation method of a low-immunogenicity absorbable suture based on a tendon of a beaver, which comprises the following steps:
(1) Killing qualified beaver, peeling off tail skin, and extracting tendon;
(2) Repeatedly cleaning the extracted tendon with deionized water until the tendon is silvery white;
(3) Soaking the tendon material obtained in the step (2) in 75% ethanol to perform dehydration fixation;
(4) Treating the tendon material obtained in the step (3) with NaOH solution, then repeatedly washing with deionized water to remove NaOH residues,
(5) Soaking the tendon material obtained in the step (4) in pepsin solution to partially remove collagen terminal peptide, then repeatedly washing with deionized water to remove pepsin solution residue,
(6) Soaking the tendon material obtained in the step (5) in glutaraldehyde solution, then repeatedly cleaning with deionized water to remove glutaraldehyde residues,
(7) Soaking the tendon material obtained in the step (6) in 75% ethanol for a second dehydration fixation, and then transferring to a newly prepared 75% ethanol solution for preservation;
(8) And (3) carrying out wire arrangement, wire branching and sterilization on the tendon material to obtain the low-immunogenicity absorbable suture based on the beaver tendon.
Further, in step (3), the soaking is performed for 24 to 36 hours.
Further, in the step (4), the concentration of the NaOH solution is 1mol/L, and the treatment is performed for 20 to 60 minutes.
Further, in step (5), the partial removal of the collagen telopeptide comprises removing 41-55% of the telopeptide.
Further, in the step (5), the concentration of pepsin in the pepsin solution is 70-90IU/mL, the pH of the pepsin solution is 2-3, and the soaking is performed at 20-22 ℃ for 36-48 hours.
Further, in the step (5), the feed liquid ratio of the tendon material to the pepsin solution is 1:40 (w/v).
Further, in step (6), the glutaraldehyde concentration in the glutaraldehyde solution is 0.1 to 0.2wt%.
Further, in step (6), the soaking is performed for 0.5 to 1 hour.
Further, in step (7), the soaking is performed for 24 to 36 hours.
Further, in step (8), the sterilization is cobalt 60 irradiation sterilization.
The beneficial effects of the invention are that
The present invention treats isolated beaver tendon with an acid enzyme method to selectively hydrolyze telopeptide of collagen, which is known in the art as the most cause of collagen to induce immune reaction, thereby reducing immunogenicity of suture based on beaver tendon, and simultaneously treats isolated beaver tendon with glutaraldehyde as a crosslinking agent at a specific concentration to increase strength of tendon, thereby improving tensile strength of suture.
It should be noted, however, that these two approaches employed by the present invention are conflicting approaches in the knowledge of the prior art. Because, in order to reduce the immunogenicity of collagen, the art will generally remove the telopeptide of collagen, whereas in order to increase the mechanical strength of the collagen biomaterial, the telopeptide should be preserved, in order to preserve the cross-linking sites so that its mechanical strength can be optimized.
As demonstrated in the examples of the present invention, in order to reduce the immunogenicity of the suture, it is necessary to increase the removal rate of the terminal peptide of the suture, but this necessarily reduces the mechanical strength of the suture. Therefore, in order to improve the mechanical strength of the suture, the invention innovatively removes partial terminal peptide from the tendon material and treats the tendon material by using glutaraldehyde solution with specific concentration, thereby realizing lower immunogenicity of the suture and ensuring that the suture has enough mechanical strength
Therefore, the present inventors have found a method for producing a suture based on a tendon of beaver, which has the immunogenicity and mechanical strength with the best overall properties, by a compromise between the two, through a lot of experiments. Specifically, the invention determines the proper treatment time for partial end peptide removal of tendon materials under specific conditions by pepsin solution at a specific feed-to-liquid ratio, and determines the proper concentration of glutaraldehyde as a cross-linking agent for enhancing the mechanical properties of tendon materials under partial end peptide removal conditions.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Example 1
The embodiment provides a low-immunogenicity absorbable suture based on a tendon of a beaver, and the preparation method comprises the following steps:
(1) Qualified quarantine, bleeding 5 adult healthy beaver mice with weight of 4-4.5kg and 12 months old, killing, taking tail of the beaver mice, sterilizing with 75% ethanol solution, carefully cutting tail skin, peeling tail skin by rotary method, and extracting tendon tissue material in clean room; the method comprises the steps of carrying out a first treatment on the surface of the
(2) Repeatedly cleaning the extracted tendon with deionized water until the tendon is silvery white;
(3) Soaking the tendon material obtained in the step (2) in 75% ethanol for 30 hours to perform dehydration fixation;
(4) Treating the tendon material obtained in the step (3) with 1mol/LNaOH solution for 30min, then washing with 10 times of 25 ℃ deionized water relative to the tendon material for 4 times, each time for 20 min, removing NaOH residues,
(5) Soaking the tendon material obtained in the step (4) in a solution containing 70IU/mL pepsin (pH is adjusted to 2.0 with glacial acetic acid) at a feed liquid ratio of 1:40 (w/v) for 48 hours at 20 ℃ to partially remove collagen telopeptide, then repeatedly washing with deionized water to remove pepsin solution residues,
(6) Soaking the tendon material obtained in the step (5) in 0.1wt% glutaraldehyde solution for 30min, then repeatedly washing with deionized water to remove glutaraldehyde residues,
(7) Soaking the tendon material obtained in the step (6) in 75% ethanol for 30 hours to perform a second dehydration fixation, and then transferring to a newly prepared 75% ethanol solution for preservation;
(8) After the dehydrated tendon material is soaked in normal saline for 2 minutes, the tendon material is subjected to artificial thread trimming and naturally dried, and then the absorbable suture semi-finished product with low immunogenicity is obtained; dividing the dried low-immunogenicity absorbable suture semi-finished product into specifications according to the wire diameter and the wire length, wearing non-damaged stainless steel needles with different specifications, and packaging; sterilizing with cobalt 60 at 15kgy dose for 15 hr to obtain absorbable suture with low immunogenicity of tendon of beaver, and storing in ventilated and dry place.
Example 2
The embodiment provides a low-immunogenicity absorbable suture based on a tendon of a beaver, and the preparation method comprises the following steps:
(1) Qualified quarantine, bleeding 5 adult healthy beaver mice with weight of 4-4.5kg and 12 months old, killing, taking tail of the beaver mice, sterilizing with 75% ethanol solution, carefully cutting tail skin, peeling tail skin by rotary method, and extracting tendon tissue material in clean room; the method comprises the steps of carrying out a first treatment on the surface of the
(2) Repeatedly cleaning the extracted tendon with deionized water until the tendon is silvery white;
(3) Soaking the tendon material obtained in the step (2) in 75% ethanol for 30 hours to perform dehydration fixation;
(4) Treating the tendon material obtained in the step (3) with 1mol/LNaOH solution for 30min, then washing with 10 times of 25 ℃ deionized water relative to the tendon material for 4 times, each time for 20 min, removing NaOH residues,
(5) Soaking the tendon material obtained in the step (4) in a solution containing 80IU/mL pepsin (pH is adjusted to 2.5 with glacial acetic acid) at a feed liquid ratio of 1:40 (w/v) for 40h at 20 ℃ to partially remove collagen telopeptide, then repeatedly washing with deionized water to remove pepsin solution residues,
(6) Soaking the tendon material obtained in the step (5) in 0.15wt% glutaraldehyde solution for 30min, then repeatedly washing with deionized water to remove glutaraldehyde residues,
(7) Soaking the tendon material obtained in the step (6) in 75% ethanol for 30 hours to perform a second dehydration fixation, and then transferring to a newly prepared 75% ethanol solution for preservation;
(8) After the dehydrated tendon material is soaked in normal saline for 2 minutes, the tendon material is subjected to artificial thread trimming and naturally dried, and then the absorbable suture semi-finished product with low immunogenicity is obtained; dividing the dried low-immunogenicity absorbable suture semi-finished product into specifications according to the wire diameter and the wire length, wearing non-damaged stainless steel needles with different specifications, and packaging; sterilizing with cobalt 60 at 15kgy dose for 15 hr to obtain absorbable suture with low immunogenicity of tendon of beaver, and storing in ventilated and dry place.
Example 3
The embodiment provides a low-immunogenicity absorbable suture based on a tendon of a beaver, and the preparation method comprises the following steps:
(1) Qualified quarantine, bleeding 5 adult healthy beaver mice with weight of 4-4.5kg and 12 months old, killing, taking tail of the beaver mice, sterilizing with 75% ethanol solution, carefully cutting tail skin, peeling tail skin by rotary method, and extracting tendon tissue material in clean room; the method comprises the steps of carrying out a first treatment on the surface of the
(2) Repeatedly cleaning the extracted tendon with deionized water until the tendon is silvery white;
(3) Soaking the tendon material obtained in the step (2) in 75% ethanol for 30 hours to perform dehydration fixation;
(4) Treating the tendon material obtained in the step (3) with 1mol/LNaOH solution for 30min, then washing with 10 times of 25 ℃ deionized water relative to the tendon material for 4 times, each time for 20 min, removing NaOH residues,
(5) Soaking the tendon material obtained in the step (4) in a solution containing 90IU/mL pepsin (pH is adjusted to 3.0 with glacial acetic acid) at a feed liquid ratio of 1:40 (w/v) for 36h at 22 ℃ to partially remove collagen telopeptide, then repeatedly washing with deionized water to remove pepsin solution residues,
(6) Soaking the tendon material obtained in the step (5) in 0.2wt% glutaraldehyde solution for 30min, then repeatedly washing with deionized water to remove glutaraldehyde residues,
(7) Soaking the tendon material obtained in the step (6) in 75% ethanol for 30 hours to perform a second dehydration fixation, and then transferring to a newly prepared 75% ethanol solution for preservation;
(8) After the dehydrated tendon material is soaked in normal saline for 2 minutes, the tendon material is subjected to artificial thread trimming and naturally dried, and then the absorbable suture semi-finished product with low immunogenicity is obtained; dividing the dried low-immunogenicity absorbable suture semi-finished product into specifications according to the wire diameter and the wire length, wearing non-damaged stainless steel needles with different specifications, and packaging; sterilizing with cobalt 60 at 15kgy dose for 15 hr to obtain absorbable suture with low immunogenicity of tendon of beaver, and storing in ventilated and dry place.
Comparative example 1
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (5), tendon material was soaked in a solution containing 80IU/mL pepsin (pH adjusted to 2.5 with glacial acetic acid) at 20 ℃ for 0h.
Comparative example 2
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (5), tendon material was soaked in a solution containing 80IU/mL pepsin (pH adjusted to 2.5 with glacial acetic acid) at 20 ℃ for 12h.
Comparative example 3
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (5), tendon material was soaked in a solution containing 80IU/mL pepsin (pH adjusted to 2.5 with glacial acetic acid) at 20 ℃ for 24h.
Comparative example 4
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (5), tendon material was soaked in a solution containing 80IU/mL pepsin (pH adjusted to 2.5 with glacial acetic acid) at 20 ℃ for 60h.
Comparative example 5
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (5), tendon material was soaked in a solution containing 80IU/mL pepsin (pH adjusted to 2.5 with glacial acetic acid) at 20 ℃ for 72h.
Comparative example 6
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, which is prepared as described in example 1, except that in step (5), the tendon material obtained in step (4) is soaked in a solution containing 70IU/mL pepsin (pH adjusted to 2.0 with glacial acetic acid) at 20 ℃ for 48 hours at a feed ratio of 1:20.
Comparative example 7
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, which is prepared as described in example 1, except that in step (5), the tendon material obtained in step (4) is soaked in a solution containing 70IU/mL pepsin (pH adjusted to 2.0 with glacial acetic acid) at 20 ℃ for 48 hours at a feed ratio of 1:30.
Comparative example 8
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, which is prepared as described in example 1, except that in step (5), the tendon material obtained in step (4) is soaked in a solution containing 70IU/mL pepsin (pH adjusted to 2.0 with glacial acetic acid) at 20 ℃ for 48 hours at a feed ratio of 1:50.
Comparative example 9
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, which is prepared as described in example 1, except that in step (5), the tendon material obtained in step (4) is soaked in a solution containing 70IU/mL pepsin (pH adjusted to 2.0 with glacial acetic acid) at 20 ℃ for 48 hours at a feed ratio of 1:60.
Comparative example 10
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (6), tendon material was soaked in 0.15wt% glutaraldehyde solution for 0min.
Comparative example 11
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that in step (6), tendon material was soaked in 0.05wt% glutaraldehyde solution for 30min.
Comparative example 12
This comparative example provides a low immunogenicity absorbable suture based on a nutria tendon, prepared as described in example 2, except that tendon material was soaked in 0.3wt% glutaraldehyde solution for 30min in step (6).
Experimental example 1
The suture lines of examples 1 to 3 and comparative examples 1 to 9 were subjected to the terminal peptide removal rate measurement, and the results are shown in Table 1 below:
table 1: end peptide removal rate of suture.
From the results of table 1, it is understood that as the soaking time of the tendon material in the pepsin solution increases, the collagen end peptide removal rate of the tendon material is higher (examples 1 to 3 and comparative examples 1 to 5); and under the same soaking time (48 h), the removal rate of the terminal peptide showed a trend of increasing and then decreasing with increasing feed-to-liquid ratio (example 1 and comparative examples 6-9). The invention uses a feed-liquid ratio of 1:40 (w/v) for subsequent experiments, with the highest removal rate of terminal peptide when the feed-liquid ratio is 1:40 (w/v).
Experimental example 2
The sutures of examples 1 to 3 and comparative examples 1 to 5 were subjected to antigenicity measurement.
Suture physiological saline solution was intravenously injected into the ear vein of rabbit every 7 days, immunized with 1ml, 2ml, 3ml, 4ml, 5ml of the progressive addition method, the negative control group was collected blood before immunization, the material group was collected 7d after the last immunization, serum was separated and ELISA test was performed. The results are shown in Table 2 below.
Table 2: serum antibody titer of rabbits after intravenous immunization.
From the results of tables 1 and 2, it can be seen that as the soaking time of the tendon material in the pepsin solution increases, the collagen end peptide removal rate of the tendon material is higher, and correspondingly the immunogenicity of the suture line is lower, which is consistent with the previous study results.
Experimental example 3
Tensile strength measurements were performed on the sutures of examples 1-3, comparative examples 1-5 and 10-12.
Diameter: the prepared tendon suture was air-dried, and the diameter was measured under a microscope by a microscopic micrometer at any 3 points (upper, middle and lower), and the average value was obtained.
Tension: one end of the suture is fixed, a weight is added to the other end of the suture until the suture breaks, and the maximum pulling force of the suture is calculated.
Tensile strength: is tensile resistance per unit cross-sectional area. According to the formula: ts=f/S, where TS represents tensile strength, F represents maximum tension to which the suture is subjected at break, and S represents suture cross-sectional area.
Table 3: mechanical properties of the suture.
| Suture diameter (mm) | Pulling force (N) | Tensile Strength (N/mm) | |
| Example 1 | 0.33 | 18.35 | 203.9 |
| Example 2 | 0.32 | 17.67 | 220.9 |
| Example 3 | 0.33 | 17.35 | 192.8 |
| Comparative example 1 | 0.32 | 21.45 | 268.1 |
| Comparative example 2 | 0.33 | 21.23 | 235.9 |
| Comparative example 3 | 0.33 | 20.64 | 229.3 |
| Comparative example 4 | 0.35 | 16.68 | 166.8 |
| Comparative example 5 | 0.34 | 15.25 | 169.4 |
| Comparative example 10 | 0.33 | 10.18 | 113.1 |
| Comparative example 11 | 0.35 | 14.36 | 143.6 |
| Comparative example 12 | 0.34 | 16.73 | 185.9 |
From the results of Table 3, it can be seen that the tensile strength of the suture tended to decrease as the removal rate of the terminal peptide of the suture increased (examples 1-3 and comparative examples 1-5). Whereas the tensile strength of the suture line tended to rise and then fall as the glutaraldehyde solution concentration increased (examples 1-3 and comparative examples 10-12), the present invention determined that the glutaraldehyde solution concentration of 0.1-0.2wt% was the optimal treatment concentration. It is noted that such an optimal treatment concentration is an optimal treatment concentration at a terminal peptide removal rate of 41-55%. As the removal rate of telopeptides increases/decreases, the optimal treatment concentration of glutaraldehyde solution will decrease/increase accordingly.
From the results of tables 1 to 3, it is found that in order to reduce the immunogenicity of the suture, it is necessary to increase the removal rate of the terminal peptide of the suture, but this necessarily reduces the mechanical strength of the suture. In order to improve the mechanical strength of the suture, the invention innovatively removes partial terminal peptide from the tendon material, and treats the tendon material by using glutaraldehyde solution with specific concentration, thereby realizing lower immunogenicity of the suture and ensuring that the suture has enough mechanical strength. According to the results of tables 1 to 3, the present invention selected a terminal peptide removal rate of 41 to 55%, at which the immunogenicity of the suture was low, and the mechanical strength of the suture obtained after treatment with glutaraldehyde solution of a specific concentration was also high, thereby allowing the suture to have the optimal combination of immunogenicity and mechanical strength.
Experimental example 4
The sutures of examples 1-3 were subjected to a systemic acute toxicity test.
The suture homogenate (suture was cut, homogenized, partially filtered and dissolved in physiological saline at a mass concentration of about 3 mg/ml) and physiological saline were injected into the abdominal cavity of each mouse, 1ml of each mouse was injected, and the body weight of the mouse was weighed at 24, 48 and 72 hours after the injection, respectively, to observe the general state, toxicity and number of dead animals of the mouse. The results show that the homogenate injected with the suture and the normal saline do not have death and poisoning reaction, and the daily weight gain of the mice is not significantly different.
Experimental example 5
The suture of example 2 was subjected to an absorbency assay.
The healthy rabbits were fixed on an operating table, shaved, anesthetized at the abdomen with procaine, the skin, peritoneum, and muscular layer were cut layer by surgical operation, the stomach, and small intestine were taken out from the abdominal cavity, and incisions each having a length of about 3cm were made, and sutured with tendon sutures (material group) and catgut (control group), respectively. The skin incision suture is covered by gauze, fixed by adhesive tape, and fed in separate cages and closely focused on the health condition of each group of animals. At 14, 25, 35, 60, 90 and 120 days, white rabbits were slaughtered, and the suture implantation site was observed to see how the suture was degraded in tension and was not absorbed. The results are shown in the following table.
Table 4: absorption rate of the suture.
From the results of Table 4, it can be seen that the suture prepared according to the present invention has good absorbability even better than the catgut.
It should be noted that while the present invention has been described in the specification with reference to the preferred embodiments thereof, the present invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but are provided to provide a more thorough understanding of the present invention. The above-described features are further combined with each other to form various embodiments not listed above, and are considered to be the scope of the present invention described in the specification; further, modifications and variations of the present invention may be apparent to those skilled in the art in light of the foregoing teachings, and all such modifications and variations are intended to be included within the scope of this invention as defined in the appended claims.
Claims (10)
1. A method for preparing a low-immunogenicity absorbable suture based on a tendon of a beaver, comprising the steps of:
(1) Killing qualified beaver, peeling off tail skin, and extracting tendon;
(2) Repeatedly cleaning the extracted tendon with deionized water until the tendon is silvery white;
(3) Soaking the tendon material obtained in the step (2) in 75% ethanol to perform dehydration fixation;
(4) Treating the tendon material obtained in the step (3) with NaOH solution, then repeatedly washing with deionized water to remove NaOH residues,
(5) Soaking the tendon material obtained in the step (4) in pepsin solution to partially remove collagen terminal peptide, then repeatedly washing with deionized water to remove pepsin solution residue,
(6) Soaking the tendon material obtained in the step (5) in glutaraldehyde solution, then repeatedly cleaning with deionized water to remove glutaraldehyde residues,
(7) Soaking the tendon material obtained in the step (6) in 75% ethanol for a second dehydration fixation, and then transferring to a newly prepared 75% ethanol solution for preservation;
(8) And (3) carrying out wire arrangement, wire branching and sterilization on the tendon material to obtain the low-immunogenicity absorbable suture based on the beaver tendon.
2. The method of claim 1, wherein in step (3), the soaking is performed for 24-36 hours.
3. The method according to claim 1, wherein in step (4), the concentration of the NaOH solution is 1mol/L, and the treatment is performed for 20 to 60min.
4. The method of claim 1, wherein in step (5) the partial removal of collagen telopeptides comprises removal of 41-55% of the telopeptides.
5. The method according to claim 4, wherein in step (5), the pepsin concentration in the pepsin solution is 70-90IU/mL, the pepsin solution has a pH of 2-3, and the soaking is for 36-48 hours at 20-22 ℃.
6. The method according to claim 4, wherein in step (5), the ratio of tendon material to pepsin solution is 1:40 (w/v).
7. The method of claim 1, wherein in step (6), the glutaraldehyde concentration in the glutaraldehyde solution is 0.1-0.2wt%.
8. The method according to claim 1, wherein in step (6), the soaking is performed for 0.5 to 1 hour.
9. The method of claim 1, wherein in step (7), the soaking is performed for 24-36 hours.
10. The method of claim 1, wherein in step (8), the sterilization is cobalt 60 irradiation sterilization.
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