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CN110232732B - Method for making computer model of defect filling block of knee joint cartilage - Google Patents

Method for making computer model of defect filling block of knee joint cartilage Download PDF

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CN110232732B
CN110232732B CN201910378365.0A CN201910378365A CN110232732B CN 110232732 B CN110232732 B CN 110232732B CN 201910378365 A CN201910378365 A CN 201910378365A CN 110232732 B CN110232732 B CN 110232732B
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knee joint
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cartilage
computer model
cartilage defect
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CN110232732A (en
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王文
秦胜男
全大萍
刘颂
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Guangzhou Red Cross Hospital (jinan University Faculty Of Medical Science Affiliated Guangzhou Red Cross Hospital)
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/10—Processes of additive manufacturing
    • B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/30—Auxiliary operations or equipment
    • B29C64/386—Data acquisition or data processing for additive manufacturing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y10/00—Processes of additive manufacturing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y50/00—Data acquisition or data processing for additive manufacturing
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T17/00—Three dimensional [3D] modelling, e.g. data description of 3D objects
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T17/00—Three dimensional [3D] modelling, e.g. data description of 3D objects
    • G06T17/10—Constructive solid geometry [CSG] using solid primitives, e.g. cylinders, cubes
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2200/00—Indexing scheme for image data processing or generation, in general
    • G06T2200/08—Indexing scheme for image data processing or generation, in general involving all processing steps from image acquisition to 3D model generation
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2210/00—Indexing scheme for image generation or computer graphics
    • G06T2210/41—Medical

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  • General Physics & Mathematics (AREA)
  • Theoretical Computer Science (AREA)
  • Mechanical Engineering (AREA)
  • Optics & Photonics (AREA)
  • Prostheses (AREA)

Abstract

The invention relates to a method for manufacturing a computer model of a cartilage defect filling block of a knee joint, which comprises the following steps: taking a healthy rabbit knee joint, fixing, decalcifying, embedding in paraffin, and longitudinally and continuously slicing; taking a picture after each slice is subjected to safranin O staining, displaying the internal three-layer structure of the knee joint of the rabbit into different colors, and reconstructing each picture into a three-dimensional image by utilizing a computer program, wherein the three-dimensional image is mainly used for displaying the internal anatomical structure of the knee joint; taking a rabbit, performing articular cartilage defect modeling, fixing, and then scanning by micro-CT layer by layer to construct a three-dimensional image of the knee joint with the articular cartilage defect; and comparing the safranin o-stained three-dimensional image with the micro-CT three-dimensional image by using a 3D two-dimensional reconstruction program to obtain a computer model of the articular cartilage defect filling block, wherein the computer model comprises an external macroscopic structure (comprising size and shape) and an internal anatomical structure (comprising a transparent cartilage layer, a calcified cartilage layer, subchondral bone and a complex interface structure between the calcified cartilage layer and the upper layer and the lower layer) of the filling block.

Description

Method for making computer model of defect filling block of knee joint cartilage
Technical Field
The invention relates to a method for manufacturing a computer model of a knee joint cartilage defect filling block.
Background
3D printing is a technology based on a material layer-by-layer accumulation manufacturing principle as the centralized embodiment of a digital technology, is an effective means for realizing individuation and precision of various orthopedic operations, and is also a hot spot direction of the current orthopedic implant research. With the development of 3D printing technology, a high-precision surgical scheme and an implant can be customized for a patient, so that the surgery is faster and more accurate, and the success rate of complex surgery is improved.
3D printing technology has developed rapidly in recent years, particularly in orthopedics and stomatology, but the successful cases are concentrated in the printing of products with single tissue structures. Articular cartilage is complex in composition and structure compared to bone tissue alone. The joint surface is surrounded by hyaline cartilage, and is divided into hyaline cartilage layer, cartilage calcified layer and subchondral bone from outside to inside, and the layers are tightly connected with each other by specific compositions and structures and have different functions. Therefore, on the basis of the existing mature technology of 3D printing of a single orthopedic material, the problem of secondary reconstruction of a three-dimensional model of a defect part and the problem of printing fusion of various 3D printing materials need to be solved.
The 3D printing can only control the mm-cm structure, and the current technology can not control the final micron-nm structure of the printed material, namely the hole structure of the material. Different pore sizes and pore structures have been proved to induce different differentiation directions of mesenchymal stem cells and to be suitable for different cell growth. Typically a pore size of 200 μm is more suitable for the proliferation and growth of chondrocytes, whereas a pore size of 50 μm is more suitable for the proliferation and growth of osteoblasts.
Thermal Induced Phase Separation (TIPS) can be applied to many polymers that cannot be resolved by wet and dry Phase Separation methods due to poor solubility. In essence, the TIPS process utilizes a latent solvent that is a solvent at high temperatures and a non-solvent at low temperatures, and the incompatibility of losing solvent power is due to the loss of thermal energy (i.e., heat as the driving force for phase separation). Because the latent solvent is non-volatile, it needs to be extracted from the finished product with a liquid that is a solvent for the latent solvent and a non-solvent for the polymer to form the microporous structure. This method is the most commonly used method among all phase separation methods, and can be used for both polar and non-polar polymers; the method has less parameters needing to be controlled in the forming process, and the process is easy to realize stability and continuity; moreover, the microstructure formed in the TIPS process is various according to the existing material, so that the special requirements of the microporous material can be met more easily; in addition, the thermally induced phase separation method can generate the same microporous structure on the thick section, and the characteristic enables the TIPS method to have unique advantages in the aspect of preparing the large-volume tissue engineering scaffold material.
3d printing of the interface structure of the complex organization is also realized by utilizing the advantages of the 3d printing technology. The calcified cartilage layer of the articular cartilage structure is positioned between the transparent cartilage layer and the subchondral bone, is tightly connected with the transparent cartilage layer through a wavy tide line structure on the upper interface of the calcified cartilage layer, and is mutually anchored with the subchondral bone through a convex-concave uneven comb-tooth-shaped bonding line structure on the lower interface of the calcified cartilage layer; the special interface connection mode increases the connection area between the articular cartilage interfaces and simultaneously increases the connection strength. Reconstruction of this structure is almost impossible by conventional tissue engineering and current 3D printing studies are more focused on the apparent size of the stent, but less on printing complex internal structures.
Aiming at the problems, the invention combines the current research situation at home and abroad, takes the main components of the articular cartilage tissue as biological ink, and utilizes the 3D model reconstruction technology, the 3D printing technology and the bionics principle to construct a bracket which has the same component structure with the articular cartilage tissue and is integrated with host tissue, so as to obtain a replaceable material which has good repetition rate and can industrially repair articular cartilage defects.
Disclosure of Invention
The invention relates to a method for manufacturing a computer model of a cartilage defect filling block of a knee joint, which comprises the following steps:
taking a healthy rabbit knee joint, fixing, decalcifying, embedding in paraffin, and longitudinally and continuously slicing;
step two, carrying out different safranin O staining on the slices obtained in the step one, so that the transparent cartilage layer, the calcified cartilage layer and the subchondral bone layer of the knee joint of the rabbit show different colors;
thirdly, photographing the slices in the second step, and after photographing, performing three-dimensional synthesis on all the two-dimensional color photographs to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint;
taking a rabbit, carrying out articular cartilage defect modeling until the subchondral bone defect is reached, taking materials, fixing, and scanning layer by layer through micro-CT to construct a three-dimensional image of the knee joint with the articular cartilage defect, wherein the three-dimensional image is used for displaying the external macroscopic structure of the knee joint with the articular cartilage defect;
and step five, comparing the three-dimensional images in the step three and the step four by using a 3D secondary model reconstruction program to obtain a computer model of the knee joint cartilage defect filling block.
Preferably, in step one, 10% neutral formaldehyde is used for fixing for 12 hours, 10% formic acid is decalcified, and after decalcification is completed, conventional paraffin embedding is carried out.
Preferably, in step one, each slice has a thickness of 5 μm.
Preferably, in step two, the hyaline cartilage layer in each section is stained red, the calcified cartilage layer is stained light blue, and the subchondral bone layer is stained blue.
Preferably, in step three, firstly, each two-dimensional color photograph is digitally shaped in sequence to obtain a gray-scale photograph with a set resolution or size; storing the gray-scale picture into a number 1 data file; the conversion system calls the data file No. 1 and performs interpolation operation to complete displacement conversion, and after each displacement conversion is completed, newly obtained data are stored in the data file No. 2; and after the storage is finished for the specified displacement times, the conversion system performs three-dimensional conversion on the No. 2 data file to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint.
Preferably, in step five, the computer model of the cartilage defect patch of the knee joint comprises an external macroscopic structure and an internal anatomical structure, the internal anatomical structure comprising a hyaline cartilage layer, a calcified cartilage layer and subchondral bone, and an interface structure between different structural layers.
The innovation point of the patent lies in that the advantages of simultaneous printing of a plurality of nozzles by combining a computer three-dimensional model reconstruction technology and a 3D printing technology are combined, and the bracket which is completely consistent with an external macro structure and a three-dimensional internal anatomical structure (comprising a transparent cartilage layer, a calcified cartilage layer and subchondral bone and an interface structure between the calcified cartilage layer and an upper layer and a lower layer) of the articular cartilage tissue defect filling block is printed. Collagen is selected as main biological ink, different materials are added into different structural layers according to components, and component bionics is achieved. After 3D printing, the adhesion or the chelation among different structural layers is not needed to be carried out by a chemical or optical method like the traditional 3D printing.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention relates to a method for manufacturing a computer model of a knee joint cartilage defect filling block, which comprises the following steps:
taking a healthy rabbit knee joint, fixing, decalcifying, embedding in paraffin, and longitudinally and continuously slicing.
In the first step, 10% neutral formaldehyde is used for fixing for 12 hours, 10% formic acid is used for decalcification, after the decalcification is completed, redundant tissues are cut off after the decalcification, articular cartilage surfaces and partial subchondral bone are reserved, normal paraffin embedding is carried out, about 300 pieces of slices are longitudinally and continuously cut, and the thickness of each slice is 5 microns.
And step two, carrying out different safranin O staining on the slices obtained in the step one, so that the transparent cartilage layer, the calcified cartilage layer and the subchondral bone layer of the knee joint of the rabbit show different colors.
In step two, the hyaline cartilage layer in each slice is stained red, the calcified cartilage layer is stained light blue, and the subchondral bone layer is stained blue.
Thirdly, photographing the slices in the second step, and after photographing, performing three-dimensional synthesis on all the two-dimensional color photographs to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint;
in the third step, firstly, each two-dimensional color picture is digitally shaped in sequence to obtain a gray-scale picture with set resolution or size; storing the gray-scale picture into a No. 1 data file; the conversion system calls the data file No. 1 and performs interpolation operation to complete displacement conversion, and after each displacement conversion is completed, newly obtained data are stored in the data file No. 2; and when the specified displacement times are stored, the conversion system performs three-dimensional conversion on the No. 2 data file to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint.
The three-dimensional image obtained by the method has the condition that tissues are distributed in a staggered mode to a large extent, the characteristic that color distinguishing is obvious according to three tissues of articular cartilage is obtained, an independent model contour curve of the distribution of the tissues is obtained, the system judges the organization of a certain region according to the attribute of a pigment point, and for the accurate separation of adjacent positions of different tissues, a polygon approximation method and a B spline curve approximation method are used for achieving the purpose.
An approximation algorithm: the title objective function is constructed by taking the sum of the squares of the distances between a point C (UK) on the approximation curve C (u) with the parameter value UK (k =0,1, \8230;, m) and the corresponding data point Qk on the contour:
Figure BDA0002052473280000041
the approximation process is as follows: cubic B-spline curves (p = 3) are C2 continuous at the nodes, sufficient to satisfy the requirements for describing contour curves, and the program will approximate with cubic periodic B-spline curves to achieve the realization of independent contours for each tissue.
And fourthly, taking the rabbit, performing articular cartilage defect modeling until the subchondral bone defect exists, taking materials, fixing, and performing micro-CT (micro-computed tomography) layer-by-layer scanning to construct a three-dimensional image of the knee joint with the articular cartilage defect, wherein the three-dimensional image is used for displaying the external macroscopic structure of the knee joint with the articular cartilage defect.
And step five, comparing the three-dimensional images in the step three and the step four by using a 3D secondary model reconstruction program to obtain a computer model of the knee joint cartilage defect filling block.
In step five, the computer model of the knee joint cartilage defect filling block comprises an external macroscopic structure and an internal anatomical structure, wherein the internal anatomical structure comprises a transparent cartilage layer, a calcified cartilage layer, subchondral bone and an interface structure between different structural layers.
The preparation method of the bionic knee joint cartilage defect filling material specifically comprises the following steps:
step one, preparation of a type I collagen solution and a type II collagen solution.
Considering that type I collagen is mainly contained in tendon and type II collagen is mainly contained in cartilage, type I and type II collagen were prepared from pig tendon and hyaline cartilage, respectively.
The type i collagen preparation process generally involves: separating pig tendon, washing with distilled water, cutting into pieces, and weighing; washing with water: washing the cut tendon with distilled water for 2-3 times, soaking in distilled water, and standing in a refrigerator at 4 deg.C for 4 hr; acid dissolution: sucking dry tendon water with filter paper, dissolving in 100 times weight volume of 0.5M acetic acid, standing in 4 deg.C refrigerator for 48 hr, and shaking continuously; salting out: centrifuging the acid solution (10000 rpm × 30min × 4-8 deg.C), adding the supernatant into an equal volume of 10% sodium chloride solution, and salting out in a refrigerator at 4 deg.C overnight (discarding the rest precipitate); and (3) acid dissolution: centrifuging the salting-out solution (4000 rpm × 10Min × 4-8 deg.C), adding 6 times volume of 0.5M acetic acid into the precipitate, performing acid dissolution again, and dissolving in a refrigerator at 4 deg.C for 4 hr to obtain collagen extractive solution; and (3) dialysis: precipitating the collagen extractive solution at high speed and low temperature, collecting the upper layer collagen solution, dialyzing with dialysis bag until pH is 5.5, subpackaging, and storing at-30 deg.C.
The process for preparing type II collagen generally comprises: cutting hyaline cartilage, slicing, defatting, mashing, homogenizing, stirring with 10 times volume of 4M guanidine hydrochloride (pH 7.5) for 24 hr, centrifugal separation, washing the precipitate, enzymolyzing with pepsin under acidic condition for 24-28 hr, centrifuging, EDTA enzymolysis of the supernatant, salting out with NaCL, dissolving with acetic acid, neutralizing with NaOH, regulating the collected collagen solution to acidity, salting out, acid dissolving, and desalting via dialysis to obtain type II collagen solution.
And step two, concentration and determination of the type I collagen solution and the type II collagen solution.
The collagen type I solution and the collagen type II solution are concentrated by polyethylene glycol.
The method specifically comprises the following steps: the extracted collagen solution was filled into dialysis bags of about 12cm in length and about 5cm in diameter by 10ml each. Sealing, placing in a beaker, pouring polyethylene glycol (molecular weight 6000) powder to cover the whole dialysis bag, concentrating at 4 deg.C, sucking out part of water from the collagen solution due to water absorption of polyethylene glycol, collecting concentrated collagen solution after 1.5 hr, measuring protein concentration, adding weak acid to adjust pH, and concentrating with polyethylene glycol again until reaching desired concentration. Collagen solutions of 5mg/ml,10mg/ml and 15mg/ml were obtained after acid-dissolution-concentration several times.
Total protein content in the type I collagen solution and the type II collagen solution was measured by the bca method.
And step three, selecting I type collagen solution and II type collagen solution with different concentrations to prepare biological ink, and simulating main components of a transparent cartilage layer, a calcified cartilage layer and a subchondral bone layer of the knee joint to add different substances.
Because the knee joint is divided into three structures of hyaline cartilage, calcified cartilage layer and subchondral bone from the joint surface to the deep part, the boundary between the structures of each layer is clear.
Since hyaline cartilage is mainly composed of chondrocytes and extracellular matrices such as type II collagen and proteoglycan, type II collagen solution (concentration 5 mg/ml) was selected as bio-ink for hyaline cartilage.
The calcified cartilage layer mainly comprises a small amount of chondrocytes, collagen type II, hydroxyapatite and other extracellular matrixes, a collagen type II solution (with the concentration of 10 mg/ml) and hydroxyapatite are directly mixed to serve as biological ink, and the mass ratio of the collagen type II solution to the hydroxyapatite is 3.
The subchondral bone layer mainly comprises osteoblasts, collagen I, hydroxyapatite and other extracellular matrixes, the subchondral bone layer is prepared by directly mixing collagen I solution (with the concentration of 15 mg/ml) and hydroxyapatite to serve as bio-ink, and the mass ratio of the collagen I solution to the hydroxyapatite is 1.
Because the main components of the transparent cartilage layer, the calcified cartilage layer and the subchondral bone layer of the simulated knee joint adopt different biological inks, different structural layers in the controllable bracket have different components.
And fourthly, performing 3D printing according to a pre-constructed computer model of the knee joint cartilage defect filling block with an internal anatomical structure (a transparent cartilage layer, a calcified cartilage layer, a subchondral bone layer and an interface structure) and an external macroscopic structure, and spraying and printing biological ink to an ultralow-temperature copper plate layer by layer to obtain a frozen block with the appearance consistent with the appearance of the filling block computer model.
The multi-nozzle printing is adopted, the biological ink with different structural layers is added into the ink box, the precision of the needle head is 100 mu m, and different biological ink can be selected for different structural layers according to different nozzles.
During printing, biological ink is extruded on an ultralow temperature copper plate with the temperature of-40 ℃ according to the instruction of a computer, and the biological ink can be frozen immediately when the thickness of each layer of biological ink sprayed by a spray head is not more than 100 mu m because the temperature of the copper plate is extremely low.
If different biological ink exists in the same layer, the biological ink is ejected through different needles, the next layer is printed after the printing is finished, the interface structure between the mm-mum support structure and different structural layers is controlled through 3D printing until the printing is finished, and finally the freezing block consistent with the external macro structure of the filling block is printed.
Because the computer constructs the internal anatomy model in advance, three different structural layers can be printed; because different structural layers adopt different biological inks, the different structural layers of the printed filling-up material have three different components.
Step five, freeze-drying the printed frozen block, wherein different pore sizes can be obtained after freeze-drying due to the selection of different collagen concentrations, and the higher the collagen concentration is, the smaller the pore diameter formed after vacuum freeze-drying is, and vice versa; finally, the bionic filling material with different structural layers, different three hole sizes and different components for knee joint cartilage defect can be prepared for filling knee joint defect.
By means of thermally induced phase separation technology, collagen with different pore sizes in different concentrations is utilized to control the pore size structure of micron-nm.
And (3) placing the printed frozen block in a refrigerator at the temperature of-80 ℃ for continuously freezing for 24 hours, then placing the frozen block into a vacuum freeze dryer for freeze drying for 24 hours by utilizing a thermally induced phase separation principle, sublimating water in the printed frozen block, directly subliming the water into gas in the freeze-drying process, forming hole structures with different sizes and apertures due to different collagen concentrations and different water contents, and finally preparing the bionic knee joint cartilage defect filling material with different hole sizes.
The bionic prosthetic materials for knee joint cartilage defect have a hole structure with mutually communicated inner parts, the higher the collagen concentration is, the smaller the pore diameter of the inner part of the prosthetic materials after freeze-drying is, and the prosthetic materials have an external macroscopic knee joint cartilage defect block structure, an internal layered structure and an interface structure between different structural layers.
Because the knee joint comprises three different structures, each structure has different components, different hole sizes and an interface structure similar to that between normal structure layers, the differentiation of the mesenchymal stem cells of the host towards different directions is facilitated, and the migration and adhesion of the chondrocytes, fibrochondrocytes and osteoblasts of the host towards the scaffold material are facilitated.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims (6)

1. A method for making a computer model of a knee joint cartilage defect filling block is characterized by comprising the following steps:
taking a healthy rabbit knee joint, fixing, decalcifying, embedding in paraffin, and longitudinally and continuously slicing;
step two, carrying out different safranin O staining on the slices obtained in the step one, so that the transparent cartilage layer, the calcified cartilage layer and the subchondral bone layer of the knee joint of the rabbit show different colors;
thirdly, photographing the slices in the second step, and after photographing, performing three-dimensional synthesis on all the two-dimensional color photographs to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint;
fourthly, taking the rabbit, performing articular cartilage defect modeling until the cartilage bone defect exists, taking materials, fixing, and performing micro-CT (micro-computed tomography) layer-by-layer scanning to construct a three-dimensional image of the knee joint with the articular cartilage defect, wherein the three-dimensional image is used for displaying the external macroscopic structure of the knee joint with the articular cartilage defect;
and step five, comparing the three-dimensional images in the step three and the step four by using a 3D secondary model reconstruction program to obtain a computer model of the knee joint cartilage defect filling block.
2. The method for preparing a computer model of a cartilage defect patch for a knee joint according to claim 1, wherein in step one, 10% neutral formaldehyde is used for fixation for 12 hours, 10% formic acid is used for decalcification, and after complete decalcification, conventional paraffin embedding is carried out.
3. The method for producing a computer model of a cartilage defect patch for a knee joint according to claim 1, wherein in the step one, each slice has a thickness of 5 μm.
4. The method for producing the computer model of the knee joint cartilage defect patch according to claim 1, wherein in step two, the hyaline cartilage layer in each slice is dyed red, the calcified cartilage layer is dyed light blue, and the subchondral bone layer is dyed blue.
5. The method for making a computer model of a defect patch for cartilage in knee joint according to claim 1, wherein in step three, each two-dimensional color photograph is digitally reshaped in sequence to obtain a grayscale photograph with a set resolution or size; storing the gray-scale picture into a number 1 data file; the conversion system calls the data file No. 1 and carries out interpolation operation to complete displacement conversion, and after each displacement conversion is completed, newly obtained data are stored in the data file No. 2; and after the storage is finished for the specified displacement times, the conversion system performs three-dimensional conversion on the No. 2 data file to obtain a three-dimensional image for displaying the internal anatomical structure of the knee joint.
6. The method for producing the computer model of the knee joint cartilage defect patch according to claim 1, wherein in step five, the computer model of the knee joint cartilage defect patch contains an external macroscopic structure and an internal anatomical structure, and the internal anatomical structure includes a transparent cartilage layer, a calcified cartilage layer and subchondral bone, and an interface structure between different structural layers.
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