CN109009550A - The evaluation method of animal model constructing method, skeletonization drug or material - Google Patents
The evaluation method of animal model constructing method, skeletonization drug or material Download PDFInfo
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- CN109009550A CN109009550A CN201811016506.6A CN201811016506A CN109009550A CN 109009550 A CN109009550 A CN 109009550A CN 201811016506 A CN201811016506 A CN 201811016506A CN 109009550 A CN109009550 A CN 109009550A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61D1/00—Surgical instruments for veterinary use
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The evaluation method of a kind of animal model constructing method, skeletonization drug or material, belongs to biomedicine field.Animal model constructing method is damaged for analog representation mankind knee subchondral bone.Construction method include: by test animal anaesthetize sb. generally processing and fixation of lying on the back；To progress preserved skin processing at the side knee joint of the test animal；Longitudinal cut is carried out centered on knee joint, and is cut on the inside of kneecap with quadriceps muscle of thigh gap, turns up kneecap to manifest articular cavity and tibial plateau articular surface；Cylindrical cavity is longitudinally formed by the center of the tibial plateau articular surface using corneal trephine.Using the rabbit knee subchondral bone injured animal model can effectively to skeletonization drug, perhaps osteogenic materials be evaluated and then provide reliable basis for the screening and evaluation of skeletonization drug or osteogenic materials.
Description
Technical field
The present invention relates to biomedicine fields, in particular to a kind of animal model constructing method, skeletonization drug or material
The evaluation method of material.
Background technique
Knee osteoarthritis (Knee Osteoarthritis, KOA) animal model is varied, including animal origin and
Production method.The reproducibility of animal model is illustrated for OA (Osteoarthritis, osteoarthritis) disease mechanisms to pass
It is important, it is also more conducive to experimental result being applied to clinic as soon as possible.
In common OA model, in addition to animal selection difference (mouse, rat, guinea pig, rabbit, dog, goat, horse etc.) it
Outside, preparation method is also to have their own characteristics each.Currently, the preparation method of OA model mainly has:
1, it spontaneously forms.Conventional animal includes: muroid, dog, horse, such as: C57/BI6, Balb/c mouse, treadmill training
Rat, guinea pig etc..Evaluation: it is clinically relevant, but disease form and disease incidence are at a distance of larger.
2, chemical induction.Conventional animal includes: muroid, rabbit, dog, horse.Method: papain, clostridiopetidase A, serum, thin
Intracellular cytokine etc..Evaluation: being difficult to apply to the mankind, easily causes articular chondrocyte apoptosis and local inflammation reaction.
3, operation induction.Conventional animal includes: muroid, rabbit, sheep, dog, horse etc..Method: anterior cruciate ligament-transection, inside pair
Ligament cutting, medial meniscus is extractd and above method combined injuries.Disadvantage: progression of disease is slow, the cartilage damage of moderate
Wound, it is highly relevant with clinic, it is not accompanied by joint destabilization, wound modeling need to be carried out.
However it is existing be related to the animal model of knee subchondral bone damage and KOA, be merely illustrative relationship-between the two
Subchondral bone damage is in close relations with KOA's, can lead to the generation of KOA.- but all about subchondral bone damage at present
Animal model is not directed to treatment, let alone induces part regeneration.
Summary of the invention
For improve, even solve the problems, such as it is in the prior art at least one, the invention proposes a kind of buildings of animal model
The evaluation method of method, skeletonization drug or material.
The present invention is implemented as follows:
In a first aspect, the embodiment of the present invention provides a kind of construction method of animal model.The structure of the animal model
Construction method is damaged for analog representation mankind knee subchondral bone.
Construction method includes:
By test animal anaesthetize sb. generally processing and fixation of lying on the back；
To progress preserved skin processing at the side knee joint of the test animal；
Carry out longitudinal cut centered on knee joint, and cut on the inside of kneecap with quadriceps muscle of thigh gap, turn up kneecap with
Manifest articular cavity and tibial plateau articular surface；
Cylindrical cavity is longitudinally formed by the center of the tibial plateau articular surface using corneal trephine.
In other one or more examples, test animal includes new zealand rabbit.
In other one or more examples, the method for general anesthesia test animal includes:
Auricular vein injection is carried out to test animal using anesthetic.
In other one or more examples, anesthetic is 4% yellow Jackets, the injection of every kilogram of test animal
Amount is 1.0mL.
In other one or more examples, the length 4cm of longitudinal cut.
In other one or more examples, the diameter of cylindrical cavity is 4mm, depth 8mm.
In other one or more examples, circle is longitudinally formed by the center of tibial plateau articular surface using corneal trephine
The method of cylindrical bore includes:
Corneal trephine is hit exactly into longitudinal screw-in 2mm by tibial plateau articular surface, and by corneal trephine together with the cartilage in it
And/or/bone tissue takes out together；
It is taken out after being inwardly further deeply no more than 8mm to total depth using low speed bone drill connection diameter 4mm drill bit, then will
Corneal trephine, which screws in, takes out the bone bolt in it.
In second aspect, the embodiment of the invention provides the evaluation methods of a kind of skeletonization drug or material.
The evaluation method of skeletonization drug or material includes:
It constructs to obtain for the damage of analog representation mankind's knee subchondral bone according to the construction method of above-mentioned animal model
Knee subchondral bone injured animal model；
Skeletonization drug or material are applied to knee subchondral bone injured animal model；
Muscle reset is carried out, discontinuity skin suture closes wound.
In other one or more examples, the method for application skeletonization drug is method that is oral, applying osteogenic materials
For minimally invasive injection.
In other one or more examples, in postoperative 4,8 weeks row MicroCT knee joint scans and X-ray film images,
To observe the regeneration situation of subchondral bone.
The utility model has the advantages that
The animal model constructing method provided through the embodiment of the present invention can obtain can be with accurate simulation knee subchondral bone
The animal model of damage.It further loses, by applying skeletonization drug or material to the animal model, observes the bone of animal model
Physiological status, thus can reach effectively to skeletonization drug perhaps osteogenic materials evaluate so that for skeletonization drug or at
The screening and evaluation of bone material provide reliable basis.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the flow diagram of the method for the building rabbit knee subchondral bone injured animal model in the embodiment of the present invention 1.
Fig. 2 shows carried out using MicroCT to the knee subchondral bone after the application osteogenic materials in the embodiment of the present invention 2
Scan the Regeneration and Repair situation obtained.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
Every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
In the present invention, in the case where not contradiction or conflict, all embodiments, embodiment and feature of the invention
It can be combined with each other.In the present invention, conventional unit, component etc., both can be commercially available, can also be public according to the present invention
The content self-control opened.In the present invention, in order to protrude emphasis of the invention, to some conventional operations and unit, component
The omission of progress, or be only briefly described.
The existing animal model applied to knee osteoarthritis is usually present different disadvantages, and especially it is merely capable of obtaining
There are certain relationships between the damage of knee subchondral bone and the animal model of KOA.But it can not but be used to evaluate and be used for
Treat the skeletonization drug of subchondral bone damage or the effectiveness of material.
In view of this, inventors herein proposing a kind of new animal model, knee subchondral bone can not only be accurately reacted
Damage, and can also be used to evaluate skeletonization drug or material, so that the research and development and production for skeletonization drug provide
More accurate, true data.
In some examples of the invention, embodiment proposes a kind of building of (rabbit) knee subchondral bone injured animal model
Method.
In embodiment, which includes:
(1) to test with new zealand rabbit anaesthetize sb. generally processing and fixation of lying on the back.
In example of the invention, to test with new zealand rabbit anaesthetize sb. generally processing and fixation of lying on the back.Some optional
Specific embodiment in, can be anaesthetized sb. generally according to the anaesthetic that the weight of new zealand rabbit gives suitable dosage.That is, anesthesia institute
The drug and its dosage of use can be selected according to the type of test animal and weight.
In the present embodiment, test animal is new zealand rabbit, therefore, according to a particular embodiment of the invention, general anesthesia
Processing can specifically be carried out according to the following steps: carry out auricular vein injection 4% according to the metering of 1.0mL/kg to new zealand rabbit
Yellow Jackets.It is possible thereby to effectively anaesthetize sb. generally to new zealand rabbit, and maintain the effective predetermined time.Wherein,
4% yellow Jackets are provided and use as anesthetic.
It should be pointed out that test animal be other than it can be adopted to new zealand rabbit, it can also be according to the need of practice
Select other test animals.In addition, anesthetic can also be selected on demand according to different animals or special demand
It selects.Alternatively, yellow Jackets may alternatively be other existing or preparations in test with the improvement of anesthetic effectiveness.
(2) to progress preserved skin processing at the side knee joint of new zealand rabbit.
Preserved skin refers in the corresponding site shaving hair removal hair for needing to carry out surgical procedure and carries out the clean operation of body surface and prepare,
It is that the clean work of operative region is carried out in the preoperative to the patient for planning to implement surgical operation.Preserved skin processing can be not only to remove body
Hair can also include the cleaning of skin, sometimes preoperative also to do skin Iodophor scouring etc..
In some surgical procedures, preserved skin can also be the non-shaving preserved skin method for being different from above-mentioned shaving operation.Wherein
Shaving preserved skin method can not be divided into depilatory agent preserved skin method again, push away mao preserved skin method and 3 kinds of detergent cleaning method.
As a kind of alternative scheme, optionally, corresponding to above-mentioned test animal-new zealand rabbit-preserved skin
Method includes: hair cutting near double knees is clean, then tincture of iodine partly sterilised 2 times, then takes off iodine disinfection 2 with 75% medicinal alcohol
It is secondary, spread aseptic hole-towel.
(3) longitudinal cut is carried out centered on knee joint, and is cut on the inside of kneecap with quadriceps muscle of thigh gap, and turn up kneecap
To manifest articular cavity and tibial plateau articular surface.
In specific alternate embodiments, operation can be carried out in the following way:
Carry out longitudinal cut centered on knee joint, and cut on the inside of kneecap with quadriceps muscle of thigh gap, turn up kneecap with
Manifest articular cavity and tibial plateau articular surface.Specific example according to the present invention, the length of notch are 4cm.
Tibial plateau articular surface not only can be effectively exposed by above-mentioned surgical procedure, but also is convenient for subsequent operation system
Make defect.In addition, finding in inventor's research, notch is too small, then the visual field is too small, is not easy to operate, and tibial plateau joint
Face exposure is incomplete；Notch is excessive, and it is serious to will lead to wound, Massive Bleeding, influences experiment and carries out.
(4) longitudinal rounding cylindrical bore is hit exactly by tibial plateau articular surface using corneal trephine, to obtain rabbit knee cartilage
Sending down the fishbone injured animal model.
As a kind of concrete implementation scheme, optionally, the diameter of cylindrical cavity is 4mm, depth 8mm.Thus shape
At appropriately sized knee subchondral bone defect, and controlling depth is 8mm to prevent from penetrating epiphyseal line.
In addition, the bone defect of the size can more efficiently evaluate skeletonization drug or osteogenic materials.Invention
People's discovery, if defect is too large or too small, the otherness that will lead between various skeletonization drugs or osteogenic materials is unobvious,
So that evaluation result does not have reliable directiveness.
As a kind of alternative example, (in figure, the tactic letter of A to E is illustrated to rabbit knee as shown in Figure 1
The orderly process being operated), above-described embodiment building rabbit knee subchondral bone injured animal model method in, system
Making cylindrical cavity can specifically carry out according to the following steps:
Corneal trephine is hit exactly longitudinal screw-in 2mm by tibial plateau articular surface by first step, and by corneal trephine together with it
Interior cartilage/bone tissue takes out together；
Second step is bored connection diameter 4mm drill bit using low speed and is inwardly further deeply no more than 8mm and taken out, then by angle
Film trepan, which screws in, takes out the bone bolt in it, plays the role of blocking defect.
It is possible thereby to safely and effectively produce bone defect, and this method accuracy is high, repeatability is high, the bone produced
Flaw size is unified, and then when carrying out the experimental evaluation of osteogenic materials multiple groups, can effectively eliminate factor shadow between animal model
It rings, improves osteogenic materials and evaluate accuracy.
Based on above animal model constructing method, in embodiments of the present invention, it is also proposed that one kind is for evaluating skeletonization
The method of drug or osteogenic materials.
In embodiment, this method (method of evaluation skeletonization drug or osteogenic materials) includes:
It is constructed to obtain rabbit knee subchondral bone injured animal model according to the method for front；
Skeletonization drug or osteogenic materials are applied to rabbit knee subchondral bone injured animal model.
According to a particular embodiment of the invention, the application skeletonization drug or osteogenic materials can be mode below:
The method of administration of skeletonization drug is oral way, and the method for administration of osteogenic materials is minimally invasive injection.Specific reality according to the present invention
Apply example, above-described embodiment for evaluating skeletonization drug or the method for osteogenic materials further comprises: in postoperative 4,8 weeks rows
MicroCT knee joint scan and X-ray film image, to observe the regeneration situation of subchondral bone.
In practice, inventors have found that by using above-mentioned building rabbit knee subchondral bone injured animal model method institute
The animal model of acquisition effectively can carry out test evaluation to a variety of skeletonization drugs or osteogenic materials, thus to skeletonization drug
Perhaps the performance of osteogenic materials product is evaluated the research and development to skeletonization drug or osteogenic materials product and provides guidance.
Obtain animal model based on above-mentioned construction method, can be effectively realized according to evaluation method to skeletonization drug or
The performance of person's osteogenic materials product is evaluated, and then provides guidance to the research and development of skeletonization drug or osteogenic materials product.
Embodiment 1: the building of animal model
(1) animal
New zealand rabbit (GB 14922-1994)；Weight: 4.5 ± 0.2kg；Gender: male and female are unlimited.
(2) instrument and material
The mating instrument of bone surgery set, product, Direct Digital flat panel X-ray imaging system (DR), general surgical procedures device
Tool, narcotics, first-aid medicine, disinfecting drug, operating room consumptive material etc..
(3) method
1. preoperative taboo is raised: the preoperative taboo for 24 hours of animal, which is raised, can't help water.
2. preoperative blank blood
2ml venous blood is acquired in auricular vein before anesthesia, for measuring blood phase and serum biochemistry.
3. anesthesia
Animal weighing, first with 4% yellow Jackets (sigma, P3761-25g；CAS.Number:57-33-0) with
1.0ml/kg dosage auricular vein injecting anesthetic.
4. preserved skin
Dorsal position is fixed, and nearby hair cutting is clean for double knees, and tincture of iodine partly sterilised 2 times, 75% medicinal alcohol takes off iodine disinfection 2
It is secondary, spread sterile wound towel, exposure art portion.
5. operation
(1) centered on knee joint, longitudinally slit skin 4cm is longitudinally slit with quadriceps muscle of thigh gap on the inside of kneecap,
Turn up kneecap, appears articular cavity；
(2) appear tibial plateau articular surface, remove the synovial membrane in the partial occlusion visual field, 4mm corneal trephine is hit exactly in platform, is indulged
To being screwed into, deep about 2mm is limited, corneal trephine is removed together and (is not abandoned) together with the cartilage/bone tissue in it.
(3) low speed brill connection 4mm drill bit inwardly further gos deep into (the not super 8mm of depth), and homemade bolt is taken to carry out again
The expansion of partial cavity, it is ensured that width 4mm, depth are 8mm (never penetrating epiphyseal line).Defect will be put in by corneal trephine before
Mouthful, the bone bolt in it is back-outed, plays the role of blocking defect.So far, subchondral bone defect model is successfully established.
Embodiment 2: the evaluation method of osteogenic materials
(1) test osteogenic materials and control osteogenic materials are inserted in subchondral bone defect point；
(2) postoperative to put row MicroCT knee joint scan and X-ray film image according to set time, to observe under cartilage
The regeneration situation of bone.Result is observed as shown in Fig. 2, proximal tibia ROI region three-dimensional reconstruction, from horizontal plane, coronal-plane, sagittal plane
Position its region, and obtain proximal tibia microcoefficient statistics such as: bone volume/total volume, bone trabecula quantity, bone trabecula are thick
Degree and bone trabecula space etc., can evaluate the effect of skeletonization drug or osteogenic materials in more detail.Convenient for quantitative analysis.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be any
It can be combined in any suitable manner in a or multiple embodiment or examples.In addition, without conflicting with each other, the technology of this field
The feature of different embodiments or examples described in this specification and different embodiments or examples can be combined by personnel
And combination.
These are only the preferred embodiment of the present invention, is not intended to restrict the invention, for those skilled in the art
For member, the invention may be variously modified and varied.All within the spirits and principles of the present invention, it is made it is any modification,
Equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of construction method of animal model damages, which is characterized in that the structure for analog representation mankind knee subchondral bone
Construction method includes:
By test animal anaesthetize sb. generally processing and fixation of lying on the back；
To progress preserved skin processing at the side knee joint of the test animal；
Longitudinal cut is carried out centered on knee joint, and is cut on the inside of kneecap with quadriceps muscle of thigh gap, turns up kneecap to appear
Articular cavity and tibial plateau articular surface out；
Cylindrical cavity is longitudinally formed by the center of the tibial plateau articular surface using corneal trephine.
2. the construction method of animal model according to claim 1, which is characterized in that the test animal includes new west
Blue rabbit.
3. the construction method of animal model according to claim 1, which is characterized in that the side of general anesthesia test animal
Method includes:
Auricular vein injection is carried out to the test animal using anesthetic.
4. the construction method of animal model according to claim 3, which is characterized in that the anesthetic is 4% penta bars of ratios
Appropriate sodium, the injection volume of every kilogram of test animal are 1.0mL.
5. the construction method of animal model according to claim 1, which is characterized in that the length 4cm of the longitudinal cut.
6. the construction method of animal model according to claim 1 or 5, which is characterized in that the cylindrical cavity it is straight
Diameter is 4mm, depth 8mm.
7. the construction method of animal model according to claim 1, which is characterized in that described to utilize corneal trephine by described
The method that the center of tibial plateau articular surface is longitudinally formed cylindrical cavity includes:
The corneal trephine is hit exactly into longitudinal screw-in 2mm by the tibial plateau articular surface, and by the corneal trephine together with it
Interior cartilage and/or/bone tissue takes out together；
It is taken out after being inwardly further deeply no more than 8mm to total depth using low speed bone drill connection diameter 4mm drill bit, then will be described
Corneal trephine, which screws in, takes out the bone bolt in it.
8. the evaluation method of a kind of skeletonization drug or material, which is characterized in that the evaluation method includes:
The construction method of animal model according to any one of claims 1 to 7 constructs to obtain for the analog representation mankind
The knee subchondral bone injured animal model of knee subchondral bone damage；
The skeletonization drug or material are applied to the knee subchondral bone injured animal model；
Muscle reset is carried out, discontinuity skin suture closes wound.
9. the evaluation method of skeletonization drug according to claim 8 or material, which is characterized in that apply the skeletonization drug
Method be it is oral, apply osteogenic materials method be minimally invasive injection.
10. the evaluation method of skeletonization drug according to claim 8 or material, which is characterized in that in postoperative 4,8 weeks rows
MicroCT knee joint scan and X-ray film image, to observe the regeneration situation of subchondral bone.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110583571A (en) * | 2019-09-16 | 2019-12-20 | 中山大学孙逸仙纪念医院 | Construction method of tendinopathy animal model |
| CN111870382A (en) * | 2020-08-07 | 2020-11-03 | 山西因美科技有限公司 | Rabbit patella subluxation operation auxiliary device |
| CN111870381A (en) * | 2020-08-07 | 2020-11-03 | 华辰未来(北京)生物医学技术有限公司 | Device capable of accelerating establishment of rabbit patella subluxation malacia model |
-
2018
- 2018-08-31 CN CN201811016506.6A patent/CN109009550A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110583571A (en) * | 2019-09-16 | 2019-12-20 | 中山大学孙逸仙纪念医院 | Construction method of tendinopathy animal model |
| CN110583571B (en) * | 2019-09-16 | 2022-11-29 | 中山大学孙逸仙纪念医院 | Construction method of tendinopathy animal model |
| CN111870382A (en) * | 2020-08-07 | 2020-11-03 | 山西因美科技有限公司 | Rabbit patella subluxation operation auxiliary device |
| CN111870381A (en) * | 2020-08-07 | 2020-11-03 | 华辰未来(北京)生物医学技术有限公司 | Device capable of accelerating establishment of rabbit patella subluxation malacia model |
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