CN113797392A - Preparation method of biological 3D printing ink containing tissue protein compound - Google Patents
Preparation method of biological 3D printing ink containing tissue protein compound Download PDFInfo
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- CN113797392A CN113797392A CN202111007548.5A CN202111007548A CN113797392A CN 113797392 A CN113797392 A CN 113797392A CN 202111007548 A CN202111007548 A CN 202111007548A CN 113797392 A CN113797392 A CN 113797392A
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Abstract
The invention provides a preparation method of biological 3D printing ink containing tissue protein complex; the preparation method comprises the following steps: step S1 tissue preprocessing: taking tissue, removing non-target components, cleaning with PBS solution, cutting the tissue into small pieces, cracking with erythrocyte lysate, centrifuging, and taking the upper layer; step S2 extraction of tissue protein complex: grinding the pretreated tissue by using a grinder, taking grinding fluid, carrying out second centrifugation, taking a water solution layer, carrying out third centrifugation after the grinding fluid is layered, taking supernate, and filtering by using a 0.22 mu m filter to obtain a tissue protein compound; step S3 prepares a biological 3D printing ink containing a tissue protein complex: mixing biological ink and tissue protein compound uniformly to obtain the final product; has the advantages that: the biological 3D printing ink prepared by the invention has high biocompatibility and biological reproducibility, meets the requirements of physiological or pathological mechanism exploration and tissue engineering repair, and has good clinical application prospect.
Description
Technical Field
The invention relates to the field of medical model manufacturing and application, in particular to a preparation method of biological 3D printing ink containing tissue protein compounds.
Background
The 3D bio-printing technology is an emerging bio-fabrication technology that mimics the microenvironment of tissues or organs in vivo to the maximum extent by using cells and specific bio-inks, thereby fabricating 3D functional living tissues. Thus, 3D bioprinting technology can be used to fabricate in vitro tissue/disease models, drug development and screening applications, understand disease progression and progression, and fabricate tissues and organs for regenerative medicine, among others.
Bio-ink is a key part of 3D bio-printing technology, and in recent years, many natural or synthetic materials with good mechanical properties, biocompatibility and tissue regeneration have emerged, such as: gelatin, sodium alginate, methacrylyl gelatin, polylactic acid, chitosan, collagen, hyaluronic acid, etc., many researchers have better simulated in vivo microenvironments by loading various bioactive factors into these synthetic and natural biomaterials.
However, since the bioactive factors abundant in the body are various in types, distributed differently and have complex action mechanisms, the microenvironment created by simply adding one or more factors is far from the real in vivo environment, and the complexity of in vivo cells and their extracellular matrix cannot be reproduced.
Disclosure of Invention
In order to solve the above problems, the present invention provides a method for preparing a bio-3D printing ink containing a tissue protein complex; the preparation method comprises the following steps:
step S1 tissue preprocessing: taking tissues, removing non-target components, cleaning with PBS (phosphate buffer solution) 2-3 times of the volume of the tissues, shearing the tissues into small pieces, cracking with erythrocyte lysate, centrifuging for the first time, and taking the upper layer to obtain pretreated tissues;
step S2 extraction of tissue protein complex: grinding the pretreated tissue by using a grinder, taking grinding fluid, carrying out second centrifugation, taking a water solution layer, carrying out third centrifugation after the grinding fluid is layered, taking supernate, and filtering by using a 0.22 mu m filter to obtain a tissue protein compound;
step S3 prepares a biological 3D printing ink containing a tissue protein complex: uniformly mixing the biological ink and the tissue protein compound in a ratio of 4:1-9:1 to prepare the biological 3D printing ink containing the tissue protein compound.
Wherein the non-target components are hair, fascia, blood vessel or fat, etc., the lipid layer is liquid, the aqueous solution layer is liquid, and the solid sediment is solid.
Further, the donor of the tissue of step S1 includes one or more of pig, mouse, rat, rabbit, or human, and the type of the tissue of step S1 includes one or more of skin, fat, liver, kidney, heart, or cartilage.
Further, the small piece of step S1 is 0.1cm3-1cm3。
Wherein the volume of the small block is determined according to tissue type, and hard tissue such as cartilage is cut into 1-2mm3The particles of (1) are cut into 0.1-1cm3Just small pieces.
Further, step S2, after grinding the pre-treated tissue with the grinder, comprises the step of adding 1ml PBS solution per gram of tissue during the grinding process.
Further, the conditions of the first centrifugation in step S1 are: centrifuging at 4 deg.C for 5min at 2000 rpm; the conditions for the first centrifugation in step S2 are: centrifuging at 4 deg.C for 5-15 min at 4000-: centrifugation was carried out at 4 ℃ for 20-40 minutes at 12000-15000 rpm.
Further, the bio-ink of step S3 is prepared from one or more of gelatin, sodium alginate, methacryl gelatin, polylactic acid, chitosan, collagen, and hyaluronic acid.
Further, the biological ink comprises the following components in percentage by mass: 10% of gelatin and 1% of sodium alginate.
The invention also provides biological 3D printing ink containing the tissue protein compound, which is prepared by the preparation method.
The invention also provides a method for preparing 3D printing tissue by using the biological 3D printing ink containing the tissue protein compound, which comprises the following steps: and (3) filling biological 3D printing ink containing a tissue protein compound into an aseptic printing cylinder, condensing, installing on a printing arm of a biological three-dimensional printer, setting the temperature to 4 ℃, adjusting the temperature of a printing platform to 0 ℃, performing three-dimensional printing on a printing nozzle with the diameter of 420 mu m in a culture dish with the diameter of 60mm to obtain a printing product, and crosslinking the printing product to obtain the 3D printing tissue.
Further, the crosslinking includes chemical crosslinking and photocrosslinking, wherein the chemical crosslinking refers to crosslinking for 10min by using 25g/l calcium chloride solution, and the photocrosslinking refers to crosslinking for 1-5min by using ultraviolet light with the wavelength of 405nm or laser or visible light with the wavelength of 632 nm.
Has the advantages that: the invention combines different materials with the obtained specific tissue protein compound, can prepare biological 3D printing ink with different requirements, for example, combines the skin or fat source protein compound with related biological ink for meeting the treatment requirement of wound surface, also can combine the cartilage source protein compound with related biological ink for meeting the bone regeneration requirement, selects different tissues and different biological inks to be arranged and combined according to different requirements, has higher biocompatibility and biological regeneration, meets the requirements of physiological or pathological mechanism exploration and tissue engineering repair, and has better clinical application prospect.
Drawings
FIG. 1 is an external view of a bio-3D printing ink containing a tissue protein complex prepared in example 1 and a general bio-ink prepared in comparative example 1;
FIG. 2 is a graph showing the mechanical properties of a general bio-ink of comparative example 1 and a bio-3D printing ink containing a tissue protein complex of example 1;
FIG. 3 is an appearance observation view of the 3D printed structure of comparative example 1 and example 1;
FIG. 4(A) is a flowchart of a method for modeling a wound surface model;
FIG. 4(B) is a diagram of an animal experiment example of successful modeling of a wound surface model of a nude mouse;
FIG. 4(C) is a view showing the wound status of each group of nude mice observed on days 2 and 8;
FIG. 4(D) is a graph showing the results of HE staining analysis of the wound surface taken from each group on day 6;
FIG. 4(E) is a statistical chart of the healing rates of the wound surfaces of the respective groups.
Detailed Description
The present invention will be further illustrated with reference to the following examples; the following examples are illustrative, not limiting, and are not intended to limit the scope of the invention; the equipment used in the invention is the equipment commonly used in the field if no special provisions are made; the methods used in the present invention are those commonly used in the art, unless otherwise specified.
Example 1
The embodiment provides a preparation method of biological 3D printing ink containing tissue protein complex, which specifically comprises the following steps:
step S1 tissue preprocessing: collecting fresh human adipose tissue discarded by liposuction surgery, washing with PBS for several times until the washing liquid is transparent and colorless, removing fascia tissue by ophthalmic scissors, and cutting adipose tissue to about 0.1cm3Cutting 10ml of the above small block-shaped adipose tissue, adding 20ml of erythrocyte lysate, lysing for 15min on ice, and collecting the lysatePouring the pretreated tissue into a centrifuge tube, tightly covering, placing into a centrifuge, centrifuging at 2000rpm and 4 deg.C for 5min, and collecting upper layer adipose tissue to obtain pretreated tissue;
step S2 extraction of tissue protein complex: and (3) placing the pretreated tissue prepared in the step (S1) in a 20ml grinder for physical grinding until the fat is completely changed into a light yellow chyme liquid, pouring the grinding liquid into a centrifuge tube, tightly covering the centrifuge tube, placing the centrifuge tube into a centrifuge, centrifuging for 10min at 4500rpm and 4 ℃, until the grinding liquid is divided into three layers, taking the middle water solution layer, pouring the middle water solution layer into the centrifuge tube again, tightly covering the centrifuge tube, placing the centrifuge tube into the centrifuge tube, centrifuging for 30min at 15000rpm and 4 ℃, taking the supernatant, and filtering by a 0.22um filter to obtain the tissue protein compound.
Step S3 prepares a biological 3D printing ink containing a tissue protein complex: preheating sterile gelatin-sodium alginate biological ink at 37 ℃ and uniformly mixing with the tissue protein compound prepared in the step S2 under the sterile condition to prepare biological 3D printing ink containing the tissue protein compound;
the embodiment also provides a 3D printing tissue, which is prepared by the following method:
taking 5ml of biological 3D printing ink containing the tissue protein compound, filling the biological 3D printing ink into a sterile printing cylinder, condensing the printing ink, installing the printing ink on a printing arm of a biological three-dimensional printer at the temperature of 4 ℃, adjusting the temperature of a printing platform to 0 ℃, performing three-dimensional printing on the printing ink in a culture dish with the diameter of 60mm, and crosslinking the printing ink for 10min by using 6ml of calcium chloride solution with the concentration of 25g/l to obtain a 3D printing tissue;
the sterile gelatin-sodium alginate biological ink comprises the following components in percentage by mass: 10% of gelatin and 1% of sodium alginate.
Comparative example 1
The control example provides a preparation method of common biological ink, which specifically comprises the following steps:
taking 5ml of sterile gelatin-sodium alginate biological ink, filling the sterile gelatin-sodium alginate biological ink into a sterile printing cylinder, condensing the sterile gelatin-sodium alginate biological ink, installing the sterile gelatin-sodium alginate biological ink on a printing arm of a biological three-dimensional printer at the temperature of 4 ℃, adjusting the temperature of a printing platform to 0 ℃, performing three-dimensional printing in a culture dish with the diameter of 60mm, and crosslinking the sterile gelatin-sodium alginate biological ink for 10min by using 6ml of calcium chloride solution with the concentration of 25g/l to obtain a 3D printing tissue;
the sterile gelatin-sodium alginate biological ink comprises the following components in percentage by mass: 10% of gelatin and 1% of sodium alginate.
Test example 13D appearance observation of printed ink
The appearance of the bio-3D printing ink containing the tissue protein complex prepared in example 1 and the general bio-ink prepared in comparative example 1 were visually observed, and the test results are shown in fig. 1.
As is clear from FIG. 1, the bio-ink obtained in example 1 was slightly cloudy compared with that obtained in comparative example 1.
Test example 2 measurement of mechanical Properties
The bio-3D printing ink containing the histone complex prepared in example 1 and the common bio-ink prepared in comparative example 1 were taken, respectively, and the rheological test of the bio-ink was performed on a TADHR-3 rheometer (arags 2) equipped with 40mm parallel plates (gap width 1 mm), and the temperature was maintained at 10 ℃ by a plate-type sensor system and a dynamic frequency sweep (frequency range between 0.1 and 10 Hz) was performed at a strain of 5% during the entire measurement, in which the storage modulus (G') and the loss modulus (G ") were measured; each bio-ink was tested in triplicate and the results are shown in figure 2.
The storage modulus and the loss modulus of the common bio-ink of the comparative example 1 and the bio-3D printing ink containing the histone complex of the example 1 are shown in fig. 2, and the time points of the transition (i.e., solidification) from the liquid state to the solid state of the two bio-inks are 74.38 ± 0.58s and 76.59 ± 0.35s, respectively, calculated from the abscissa of the intersection of the storage modulus and the loss modulus at 10 ℃, and the solidification time of the 3D printing ink of the example 1 is slightly longer than that of the common bio-ink of the comparative example 1.
Test example 33D appearance Observation of printed texture
The 3D printed structures obtained in example and comparative example 1 were taken, and the appearance was visually observed, and the test results are shown in fig. 3.
As is clear from FIG. 3, the printability of example 1 was not significantly different from that of comparative example 1, and it was confirmed that the printability of the biological 3D printing ink containing the tissue protein complex of example 1 was good and the fidelity of the printed tissue was high.
Test example 4 wound treatment test
The test method is as follows:
the dorsal skin of nude mice was used as a full-thickness wound model, as shown in fig. 4 (a): covering sterile gauze on the wound surface after different treatment measures are taken, and fixing the wound surface by using an elastic bandage; opening the bandage every 2 days when changing the dressing, observing the wound surface and taking a picture, and fixing the wound surface by using the elastic bandage again after changing the sterile gauze; opening a bandage during tissue acquisition, cutting off a whole layer of skin tissue within 0.5cm from the edge of a wound surface by using an ophthalmic scissors, and soaking and fixing the whole layer of skin tissue in paraformaldehyde solution in time; before the experiment, a round skin full-thickness wound surface with the diameter of 1cm is cut on the back of each nude mouse by ophthalmology, an animal experiment example is shown in figure 4(B), model mice successfully molded are divided into a blank group, a tissue protein compound group, an experiment group and a control group, the tissue protein compound group, the experiment group and the control group are respectively applied with the tissue protein compound (100 mu g/ml solution) of example 1, the 3D printing tissue prepared in example 1 and the 3D printing tissue prepared in control example 1 on the back wound surface of the nude mouse, and the wound surface treatment effect of each group is observed; the blank group was not treated; the wound surface states of the groups of nude mice are observed on the 2 nd day and the 8 th day respectively, the test result is shown in figure 4(C), the wound surfaces of the groups are taken on the 6 th day respectively and subjected to HE staining analysis, the test result is shown in figure 4(D), data statistics is carried out on the wound surface healing rate of each time point (the 10 th day and the 14 th day), and the calculation method of the wound surface healing rate is as follows: taking a wound surface picture under the same visual field and magnification, measuring the area of the wound surface by using data processing software, wherein the ratio of the difference value of the area of the wound surface and the initial area of the wound surface at different time points to the initial area of the wound surface is the healing rate; the experimental results are shown in FIG. 4 (E).
The test results are as follows:
as can be seen from fig. 4(C), a large amount of yellowish exudate was visible on the wound surface of the blank group on day 2 to cover the wound surface; the wound surface of the tissue protein compound group is wet, and no obvious exudation is seen; the wound surfaces of the control group and the experimental group are degraded, the wound surfaces are clean and have no obvious exudation; on day 8, the blank group had large wound residue, dry and crusted wound, the tissue protein complex group had bright red wound surface but insufficient epithelization, the control group had small residual wound surface area, and the wound surface base was light pink; the area of the residual wound surface of the experimental group is obviously smaller than that of the first three groups, and the base of the wound surface is bright red.
As can be seen from fig. 4(D), the wound surface of the blank group was significantly contracted, scabs were present on the surface, vascularization and inflammatory cell infiltration were present to a certain extent under the scabs, the wound surface of the tissue-protein complex group was slightly contracted, many new blood vessels and inflammatory infiltration were visible in the wound base, the wound surface of the control group was not significantly contracted, but the wound base was not vascularized enough, the wound surface of the experimental group was not significantly contracted, and the degree of vascularization of the wound base was moderate.
As can be seen from fig. 4(E), the wound surface of the experimental group healed completely at day 10, which is significantly higher than that of the three groups (p < 0.05); on day 14, the wound surface of the blank group did not heal, and the other three groups healed completely; in conclusion, the healing speed and effect of the experimental group are obviously better than those of the control group and the tissue protein complex group.
Finally, it should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered by the claims of the present invention.
Claims (10)
1. A preparation method of biological 3D printing ink containing tissue protein complex is characterized by comprising the following steps:
step S1 tissue preprocessing: taking tissues, removing non-target components, cleaning with PBS (phosphate buffer solution) 2-3 times of the volume of the tissues, shearing the tissues into small pieces, cracking with erythrocyte lysate, centrifuging for the first time, and taking the upper layer to obtain pretreated tissues;
step S2 extraction of tissue protein complex: grinding the pretreated tissue by using a grinder, taking grinding fluid, carrying out second centrifugation, taking a water solution layer, carrying out third centrifugation after the grinding fluid is layered, taking supernate, and filtering by using a 0.22 mu m filter to obtain the tissue protein compound;
step S3 prepares a biological 3D printing ink containing a tissue protein complex: and uniformly mixing the biological ink and the tissue protein compound in a ratio of 4:1-9:1 to prepare the biological 3D printing ink containing the tissue protein compound.
2. The method of claim 1, wherein the donor of the tissue of step S1 includes one or more of pig, mouse, rat, rabbit, or human, and the type of the tissue of step S1 includes one or more of skin, fat, liver, kidney, heart, or cartilage.
3. The method of claim 1, wherein the small pieces of step S1 are 0.1cm3-1cm3。
4. The method of claim 1, wherein the step S2, after grinding the pre-treated tissue with the grinder, further comprises adding 1ml of PBS solution per gram of tissue during grinding.
5. The method of claim 1, wherein the conditions of the first centrifugation in step S1 are: centrifuging at 4 deg.C for 5min at 2000 rpm; the conditions of the first centrifugation in step S2 are: centrifuging at 4 ℃ for 5-15 minutes at 4000-: centrifugation was carried out at 4 ℃ for 20-40 minutes at 12000-15000 rpm.
6. The method of claim 1, wherein the bio-ink of step S3 is prepared from one or more of gelatin, sodium alginate, methacryl gelatin, polylactic acid, chitosan, collagen, and hyaluronic acid.
7. The preparation method of claim 6, wherein the bio-ink comprises the following components in percentage by mass: 10% of gelatin and 1% of sodium alginate.
8. A bio 3D printing ink containing a tissue protein complex, which is prepared by the preparation method of any one of claims 1 to 7.
9. A method for preparing 3D printed tissue from the biological 3D printing ink containing tissue protein complexes according to claim 8, wherein the method comprises: and (3) loading the biological 3D printing ink containing the tissue protein compound into a sterile printing cylinder, condensing, installing on a printing arm of a biological three-dimensional printer, setting the temperature to 4 ℃, adjusting the temperature of a printing platform to 0 ℃, printing the diameter of a nozzle to be 420 mu m, performing three-dimensional printing in a culture dish with the diameter of 60mm to obtain a printed product, and crosslinking the printed product to obtain the 3D printing tissue.
10. The method of claim 9, wherein the crosslinking comprises chemical crosslinking and photocrosslinking, the chemical crosslinking refers to crosslinking for 10min using 25g/l calcium chloride solution, and the photocrosslinking refers to crosslinking for 1-5min using ultraviolet light with a wavelength of 405nm or laser or visible light with a wavelength of 632 nm.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087146A (en) * | 2011-11-04 | 2013-05-08 | 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心 | Method for highly-efficiently extracting proteins of sheep skin tissues |
| US20180055643A1 (en) * | 2016-08-07 | 2018-03-01 | Nanochon, Llc | Three-dimensionally printed tissue engineering scaffolds for tissue regeneration |
| CN109957540A (en) * | 2017-12-14 | 2019-07-02 | 深圳先进技术研究院 | A kind of 3D printing adipose tissue |
| CN111166937A (en) * | 2020-01-22 | 2020-05-19 | 中国人民解放军总医院 | Acellular extracellular matrix, preparation method thereof and biological ink |
| CN111228574A (en) * | 2020-01-22 | 2020-06-05 | 中国人民解放军总医院 | Tissue engineering meniscus scaffold and preparation method thereof |
| CN111544657A (en) * | 2020-05-11 | 2020-08-18 | 北京大学第三医院（北京大学第三临床医学院） | Preparation method of cell 3D printing biological ink with good printability |
| CN111748526A (en) * | 2020-07-03 | 2020-10-09 | 中山大学 | Preparation method of extracellular matrix-imitated printable composite biological ink |
-
2021
- 2021-08-30 CN CN202111007548.5A patent/CN113797392A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087146A (en) * | 2011-11-04 | 2013-05-08 | 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心 | Method for highly-efficiently extracting proteins of sheep skin tissues |
| US20180055643A1 (en) * | 2016-08-07 | 2018-03-01 | Nanochon, Llc | Three-dimensionally printed tissue engineering scaffolds for tissue regeneration |
| CN109957540A (en) * | 2017-12-14 | 2019-07-02 | 深圳先进技术研究院 | A kind of 3D printing adipose tissue |
| CN111166937A (en) * | 2020-01-22 | 2020-05-19 | 中国人民解放军总医院 | Acellular extracellular matrix, preparation method thereof and biological ink |
| CN111228574A (en) * | 2020-01-22 | 2020-06-05 | 中国人民解放军总医院 | Tissue engineering meniscus scaffold and preparation method thereof |
| CN111544657A (en) * | 2020-05-11 | 2020-08-18 | 北京大学第三医院（北京大学第三临床医学院） | Preparation method of cell 3D printing biological ink with good printability |
| CN111748526A (en) * | 2020-07-03 | 2020-10-09 | 中山大学 | Preparation method of extracellular matrix-imitated printable composite biological ink |
Non-Patent Citations (1)
| Title |
|---|
| 付小兵 等: "《生物3D打印与再生医学》", 29 February 2020, 华中科技大学出版社 * |
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