CN112089890A - Acellular matrix hydrogel and preparation method and application thereof - Google Patents
Acellular matrix hydrogel and preparation method and application thereof Download PDFInfo
- Publication number
- CN112089890A CN112089890A CN202010889600.3A CN202010889600A CN112089890A CN 112089890 A CN112089890 A CN 112089890A CN 202010889600 A CN202010889600 A CN 202010889600A CN 112089890 A CN112089890 A CN 112089890A
- Authority
- CN
- China
- Prior art keywords
- transferring
- acellular
- stirring
- acellular matrix
- solution containing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
Abstract
The application discloses a preparation method of acellular matrix hydrogel, which comprises the following steps: obtaining a tissue mass; transferring the tissue block into a solution containing 0.02% of pancreatin and 0.05% of ethylenediamine tetraacetic acid, and stirring for 2-3 h; then transferring the tissue block into a solution containing 3% Triton X, and stirring for 2-3 h; then transferring the tissue block into a solution containing 4% sodium deoxycholate and stirring for 2-3 h; moving the tissue block into deionized water to be soaked for 12-18 h, and obtaining a decellularized scaffold; transferring the acellular scaffold into 75% alcohol, soaking for 30min, and transferring into a solution containing 0.1% peracetic acid and stirring for 2-3 h; freeze-drying and grinding the acellular scaffold into matrix powder, and then transferring the matrix powder into a solution containing pepsin hydrochloride, stirring and digesting for 2-3d to obtain a pre-gel; and diluting the pre-gel to a preset concentration, and standing to obtain the acellular matrix hydrogel. The application can efficiently extract the acellular matrix hydrogel from the umbilical cord tissue, and furthest reserve the integrity of the internal structure of the acellular matrix.
Description
Technical Field
The application relates to the field of biological materials, in particular to an acellular matrix hydrogel and a preparation method and application thereof.
Background
In vivo, cellular turnover and differentiation are coordinated with a constantly changing environment characterized by a spatio-temporal gradient of various factors through interactions with neighboring cells, which is difficult to achieve under standard two-dimensional culture conditions. The development of 3D culture systems that mimic the natural tissue microenvironment has become a research hotspot. Extracellular matrix (ECM) is a complex secreted extracellularly by cells of tissues and organs, and is distributed on the cell surface and in the intercellular spaces. The composition is complex, is a complex of structural protein and functional protein, and specifically comprises type I collagen, type III collagen, fibronectin, laminin, glycosaminoglycan (GAG), various cell growth factors, and the like. ECM is trypsinized to obtain ECM hydrogels, which is considered an ideal tool in all biomaterials in the field of tissue engineering.
Existing ECM hydrogels include: liver, small intestine submucosa, dermis and bladder matrix, but the healthy tissues from human ECM are very limited and relate to ethical problems, species difference exists in animal ECM sources, virus infection and immunogenicity among species are easily caused, and the artificially synthesized hydrogel has single component and poor biological tissue compatibility. The animal decellularized ECM hydrogel has the problems of ethics, limited sources, complex extraction process, low efficiency and the like due to species difference, and is difficult to be used clinically.
Disclosure of Invention
The application aims to overcome the defects of the prior art and provide the acellular matrix hydrogel with high extraction efficiency and good quality, and the preparation method and the application thereof.
In order to achieve the technical purpose, the technical scheme adopted by the application is as follows:
firstly, a preparation method of acellular matrix hydrogel comprises the following steps:
obtaining a tissue mass;
transferring the tissue block into a solution containing 0.02% of pancreatin and 0.05% of ethylenediamine tetraacetic acid, and stirring at the rotating speed of 350rmp for 2-3h at the temperature of 37 ℃;
then transferring the tissue block into a solution containing 3% Triton X, and stirring for 2-3h at the rotating speed of 250rmp at the temperature of 27 ℃;
then transferring the tissue block into a solution containing 4% sodium deoxycholate, and stirring at the rotating speed of 250rmp for 2-3h at the temperature of 27 ℃;
moving the tissue block into deionized water, and soaking for 12-18 h at 27 ℃ to obtain a decellularized scaffold;
transferring the decellularized scaffold into 75% alcohol, soaking for 30min, transferring into a solution containing 0.1% peroxyacetic acid, and stirring at the rotating speed of 150rmp for 2-3 h;
freeze-drying and grinding the acellular scaffold into matrix powder, and then transferring the matrix powder into a solution containing pepsin hydrochloride, stirring and digesting for 2-3d to obtain a pre-gel;
and (3) diluting the pre-gel to a preset concentration at 37 ℃, and standing to obtain the acellular matrix hydrogel.
Preferably, the tissue mass is derived from the umbilical cord.
Further preferably, the tissue blocks are cut into 3-4cm wide sections.
Further, before the tissue blocks and the acellular scaffolds are moved into different treatment solutions, the method further comprises the step of washing the tissue blocks and the acellular scaffolds with deionized water.
Specifically, the freeze-drying and grinding of the decellularized scaffold into a matrix powder comprises:
placing the decellularized scaffold in an environment at-80 ℃ for 3-4 h;
transferring to a freeze dryer for freeze drying treatment;
transferring the mixture into a grinder with precooling by liquid nitrogen for grinding.
Optionally, the grinding frequency of the grinder is 60Hz, and the grinding time is 120 s.
Preferably, the concentration of the pre-gel is 10 mg/ml. The pre-gel is neutralized with phosphate buffer and sodium hydroxide before being used to culture cells and prepare organoids.
Secondly, the acellular matrix hydrogel is prepared by adopting the preparation method of the acellular matrix hydrogel.
The application of the acellular matrix hydrogel in preparing a drug carrier and/or in preparing a stem cell transplantation carrier and/or in constructing an artificial organ is also disclosed.
Or the application of the preparation method of the acellular matrix hydrogel in preparing a drug carrier, and/or the application in preparing a stem cell transplantation carrier, and/or the application in constructing an artificial organ.
Compared with the prior art, the method has the following advantages:
(1) the umbilical cord is preferably used as a tissue material for preparing the acellular matrix hydrogel, the umbilical cord is used as a special attribute of human tissue medical waste, the source is sufficient, the species and ethical issues are not involved, if the acellular ECM hydrogel can be extracted from the umbilical cord, a new way for treating Mesenchymal Stem Cells (MSCs) is expected to be developed, the survival efficiency of the transplanted MSCs is obviously improved, the specificity is directed to damaged tissues, and the risk of gene mutation and tumorigenicity is reduced. Engineered organs can also be created with appropriate cell filling, showing great application prospects in the fields of tissue engineering and regenerative medicine.
(2) The application establishes a tetrad scheme of pancreatin, Triton X, sodium deoxycholate and peracetic acid, and the acellular ECM hydrogel is efficiently extracted from the umbilical cord tissue, so that the high-quality acellular scaffold can be obtained within 1-2d, the hydrogel can be obtained within 6-7d, and the integrity of the internal structure of the ECM is maintained to the maximum extent.
(3) The present application allows for clinical GMP-grade production, and chemicals and reagents used in each step of hydrogel production are commercially available in GMP-grade commercial products, facilitating industrial use.
(4) The method and the prepared acellular matrix hydrogel can be used for 3D culture systems of various stem cells and organoids.
Drawings
FIG. 1 is a physical representation of intermediate and final products obtained using the acellular matrix hydrogel production method of the present application.
FIG. 2 is a graph showing the identification of the staining of the acellular matrix hydrogel of the present application.
FIG. 3 is an identification chart of immunofluorescence staining of an acellular matrix hydrogel of the present application.
FIG. 4 is a graph of a protein mass spectrometry analysis of the acellular matrix hydrogel of the present application.
Fig. 5 is an example of the application of acellular matrix hydrogel combined with various cells to prepare a liver organoid for acute liver failure transplantation.
FIG. 6 is an example graph of a 3D culture of stem cells using the acellular matrix hydrogel of the present application.
Detailed Description
The present application is described in further detail below with reference to the attached drawings and the detailed description.
Example one
Preparation method of acellular matrix hydrogel
The umbilical cord is taken as a main treatment object in the embodiment of the application, and tissues from other sources are also applicable to the preparation method of the application. The preparation method of the acellular matrix hydrogel comprises the following steps:
(1) obtaining a tissue mass;
after obtaining fresh umbilical cord tissue, the umbilical cord tissue needs to be frozen at-20 ℃ for transportation to a laboratory, and particularly, liquid nitrogen can be used for quick freezing. In the next step, virus, bacteria and mycoplasma detection is carried out on the tissue blocks to ensure that the subsequently prepared acellular matrix hydrogel is free from microbial contamination. After confirming that the tissue mass was satisfactory, the surface blood was rinsed with deionized water and the arteries and veins of the umbilical cord were removed and ready for use.
(2) And (3) decellularizing the tissue block:
a) the cord was cut into 3-4cm pieces (see FIG. 1a) and stirred in deionized water 3 times at room temperature for 1h at 300 rmp.
b) The umbilical cord tissue is moved into a solution containing 0.02% of pancreatin and 0.05% of ethylenediaminetetraacetic acid (EDTA) and stirred for 2 hours at a stirring speed of 350rmp, and the temperature is kept at 37 ℃ all the time in the experiment.
c) The umbilical cord was washed with deionized water 3 times for 10min each time.
d) The umbilical cord is placed in a Triton X solution with 3% of content and stirred for 2 hours on a shaking table with the speed of 250rmp and the temperature of 27.0 ℃.
e) The umbilical cord was washed with deionized water 3 times for 10min each time.
f) Putting the umbilical cord into a sodium deoxycholate solution with the concentration of 4%, stirring the solution on a shaking table for 2.5 hours at the rotation speed of 250rmp and the temperature of 27.0 ℃.
g) The umbilical cord is washed by excessive deionized water for 15min and then soaked in deionized water for 12h at the temperature of 27.0 ℃.
As shown in FIG. 1b, an acellular matrix scaffold was obtained from umbilical cord tissue, which had a whitish, porous appearance.
(3) And (3) sterilizing the acellular matrix scaffold:
the subsequent operations are carried out in a clean bench.
h) The acellular matrix scaffold is soaked in 75% alcohol for 30 min.
i) The umbilical cord was washed with deionized water 3 times for 10min each time.
j) Transferring into a peroxyacetic acid solution containing 0.1% and stirring for 2-3h at the speed of 150 rpm.
k) Then transferring to 1 XPBS solution (phosphate buffer solution), stirring, soaking for 15min, replacing deionized water, and repeating for 3-5 times.
(4) Freezing, drying and grinding the acellular matrix scaffold:
a) placing the acellular matrix scaffold into a clean culture dish, paving, packaging in sterile bags, and freezing in a refrigerator at-80 deg.C for 3-4 h.
b) And after the cell-free matrix scaffold is completely frozen, putting the cell-free matrix scaffold into a freeze dryer for freeze drying. Taking out the freeze-dried acellular matrix scaffold, shearing, placing into a grinding steel tank after autoclaving, precooling for about 15min in a liquid nitrogen environment, and grinding in a grinder (60Hz and 120 s). The matrix powder was taken out of the milling steel pot (see fig. 1c and 1 d).
(5) Acellular matrix scaffold digestion:
digesting in 100ml hydrochloric acid pepsin solution for 2-3 days to obtain 10mg/ml pre-gel (see figure 1e), storing at 4 deg.C for aseptic storage, and diluting to preset concentration for subsequent use to obtain acellular matrix hydrogel.
Further, the pre-gel needs to be neutralized with 10 x PBS and NaOH before being diluted to prepare for cell culture or organoid preparation.
In conclusion, the preparation cycle from the decellularization step to the obtainment of the decellularized matrix hydrogel is not more than one week. From the reagent used, pancreatin, Triton X, sodium deoxycholate and peroxyacetic acid are taken as main components, and ribonuclease and deoxyribonuclease are not required to be used; the surfactants used are also milder. The mechanical agitation is mainly performed by a shaking table.
Example two
Standardized identification of acellular matrix hydrogels
The acellular matrix hydrogel can be clinically applied only by meeting specified physicochemical indexes. Specifically, the items and results of the standardized identification test of the acellular matrix hydrogel include:
A. gelling experiment: and (3) balancing the pre-gel at 37 ℃, standing at room temperature for 10min to obtain the hydrogel (shown in figure 1f), wherein the hydrogel is a white semitransparent three-dimensional solid gel and has extremely strong plasticity.
B. Yield: the total weight of the 3 batches of 10 umbilical cord acellular scaffolds after freeze drying is 16.6g, and 1.66g of freeze-dried powder can be obtained on average per umbilical cord after grinding, and 160ml of pre-gel can be prepared.
C. And (3) scanning electron microscopy: the sample is in a block shape, the superfine structure can be well preserved after being processed by observation under a scanning electron microscope, and the nanofiber collagen structure is intact even under high magnification (see figure 1g), so that the effectiveness of the scheme in extracting the umbilical cord acellular matrix hydrogel is proved, and the damage of the preparation method provided by the application to the acellular matrix scaffold is very small. The acellular matrix hydrogel is in a randomly oriented fiber network three-dimensional structure and mainly comprises radially arranged collagen fibers, a layered pore structure (pores in pores) of additional flexible structural proteins and fibronectin. Collagen fibers with different thicknesses are mutually interwoven to form a three-dimensional space, so that cavities with different apertures and a thick frame structure are formed, and a structural foundation is provided for cell adhesion.
And D, HE staining: hematoxylin-eosin staining (HE staining for short) is one of the staining methods commonly used in paraffin section technology. The hematoxylin staining solution is alkaline, and mainly makes the chromatin in the cell nucleus and the nucleic acid in the cytoplasm bluish; eosin is an acid dye that primarily reddens components in the cytoplasm and extracellular matrix. As shown in fig. 2 h-2 j, the acellular matrix hydrogel of the present application showed typical pink eosinophilic staining for collagen, whereas no typical basophilic staining for nuclear material was observed, indicating that the acellular matrix hydrogel of the present application was free of cells and nuclear remnants, and that organized collagen fibers, vascular pores, and also fibronectin fibers within the pore walls were visible.
Masson staining: masson staining, which refers to mixing two or three anionic dyes so that collagen fibers appear blue and muscle fibers appear red, is one of the staining methods for revealing fibers in tissues. Trichrome staining by Masson again confirmed the results as shown by HE staining, with uniform blue staining indicating collagen retention in the biological matrix as shown in fig. 2k to fig. 2 m.
F. And (3) immunofluorescence staining: immunofluorescent color was performed on the four major collagen components of the acellular matrix hydrogel and on the nuclei, which were stained with DAPI (4', 6-diamidino-2-phenylindole). As shown in fig. 3, the staining results showed staining of collagen I, IV, laminin and fibronectin, with little blue fluorescence of DAPI in the acellular matrix hydrogel after decellularization.
Determination of dsdna content; the DNA content of the natural umbilical cord sample is 5160.12 +/-284.27 ng/mg, and the DNA content of the decellularized sample is 19.42 +/-3.20 ng/mg, which shows that the nucleic acid substances in the decellularized matrix hydrogel are basically removed, and the acellular matrix hydrogel meets the requirements of experiments and clinics.
H. Protein mass spectrometry analysis: three biological replicates of the acellular matrix hydrogel of the present application were performed, for a total of 1320 proteins identified (see figure 4), and then 883 proteins were screened at a value of 2 or more according to the unique peptide assay, of which about 154 proteins were considered to be derived from ECM and 536 proteins were considered to be derived from exosomes, confirming that the acellular matrix hydrogel had a dual role as a support structure and storage of the absorbed soluble signal.
The results of the standardized identification and detection items show that the acellular matrix hydrogel obtained by the preparation method of the acellular matrix hydrogel completely retains the pore structure of the acellular scaffold, is thorough in acellular, has high quality, and is particularly suitable for preparing the acellular matrix hydrogel of the umbilical cord. The acellular matrix hydrogel of the umbilical cord is used as a safe material produced by GMP (good manufacturing practice) process, provides a safe and effective carrier for clinical in-vivo transplantation of MSCs (mesenchymal stem cells), and is expected to be used for tissue engineering and long-term culture of various stem cells and organoids (as shown in figure 5). Specifically, after the acellular matrix hydrogel is mixed with umbilical cord mesenchymal stem cells (UC-MSCs), embryonic stem cells (ES) or induced pluripotent stem cells (IPS), a cell sheet cultured in a serum-free culture medium is added for 1-3 days to obtain a 3D (three-dimensional) cell sheet (as shown in figure 6), and multiple cells are added for co-culture to obtain a corresponding organoid.
Further, the acellular matrix hydrogel may also be layered, sprayed, or otherwise applied with other scaffold materials as a drug carrier, or refilled with appropriate cells or applied as a bio-link to three-dimensional bioprinting to create an engineered organ. It is also envisioned that the carrier material can be used for various stem cell transplants.
In summary, the application utilizes the quadruple scheme of pancreatin, Triton X, sodium deoxycholate and peracetic acid to efficiently extract the acellular matrix hydrogel from the umbilical cord tissue, has short preparation time, furthest retains the integrity of the internal structure of the acellular matrix, and provides a foundation for the application of the acellular matrix hydrogel.
The above embodiments are only preferred embodiments of the present application, but not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present application should be construed as equivalents and are included in the scope of the present application.
Claims (11)
1. A preparation method of acellular matrix hydrogel is characterized by comprising the following steps:
obtaining a tissue mass;
transferring the tissue block into a solution containing 0.02% of pancreatin and 0.05% of ethylenediamine tetraacetic acid, and stirring at the rotating speed of 350rmp for 2-3h at the temperature of 37 ℃;
then transferring the tissue block into a solution containing 3% Triton X, and stirring for 2-3h at the rotating speed of 250rmp at the temperature of 27 ℃;
then transferring the tissue block into a solution containing 4% sodium deoxycholate, and stirring at the rotating speed of 250rmp for 2-3h at the temperature of 27 ℃;
moving the tissue block into deionized water, and soaking for 12-18 h at 27 ℃ to obtain a decellularized scaffold;
transferring the decellularized scaffold into 75% alcohol, soaking for 30min, transferring into a solution containing 0.1% peroxyacetic acid, and stirring at the rotating speed of 150rmp for 2-3 h;
freeze-drying and grinding the acellular scaffold into matrix powder, and then transferring the matrix powder into a solution containing pepsin hydrochloride, stirring and digesting for 2-3d to obtain a pre-gel;
and (3) diluting the pre-gel to a preset concentration at 37 ℃, and standing to obtain the acellular matrix hydrogel.
2. The method of claim 1, wherein the tissue mass is derived from umbilical cord.
3. The method of claim 1, wherein the tissue mass is cut into pieces 3-4cm wide.
4. The method of claim 1, further comprising the step of rinsing with deionized water prior to transferring the tissue mass and the decellularized scaffold into the different treatment solutions.
5. The method of claim 1, wherein lyophilizing and grinding the decellularized scaffold to a matrix powder comprises:
placing the decellularized scaffold in an environment at-80 ℃ for 3-4 h;
transferring to a freeze dryer for freeze drying treatment;
transferring the mixture into a grinder with precooling by liquid nitrogen for grinding.
6. The method according to claim 5, wherein the grinding frequency of the grinder is 60Hz and the grinding time is 120 s.
7. The method of claim 1, wherein the pre-gel is at a concentration of 10 mg/ml.
8. The method of claim 1, wherein said pre-gel is neutralized with phosphate buffer and sodium hydroxide prior to use in culturing cells and preparing organoids.
9. An acellular matrix hydrogel prepared by the method for preparing an acellular matrix hydrogel according to claims 1-8.
10. Use of the acellular matrix hydrogel of claim 9 in the preparation of a pharmaceutical carrier, and/or in the preparation of a stem cell transplant carrier, and/or in the construction of an artificial organ.
11. The use of the method of claim 1 to 8 for the preparation of a drug carrier, and/or for the preparation of a stem cell transplantation carrier, and/or for the construction of an artificial organ.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010889600.3A CN112089890A (en) | 2020-08-28 | 2020-08-28 | Acellular matrix hydrogel and preparation method and application thereof |
| PCT/CN2021/115121 WO2022042704A1 (en) | 2020-08-28 | 2021-08-27 | Decellularized matrix hydrogel, preparation method therefor, and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010889600.3A CN112089890A (en) | 2020-08-28 | 2020-08-28 | Acellular matrix hydrogel and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112089890A true CN112089890A (en) | 2020-12-18 |
Family
ID=73758214
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010889600.3A Pending CN112089890A (en) | 2020-08-28 | 2020-08-28 | Acellular matrix hydrogel and preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN112089890A (en) |
| WO (1) | WO2022042704A1 (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113082295A (en) * | 2021-04-02 | 2021-07-09 | 大连理工大学 | Derived scaffold based on skin-derived acellular matrix and construction method thereof |
| CN113881627A (en) * | 2021-10-21 | 2022-01-04 | 焕生汇生物基因技术(北京)有限公司 | Method for improving yield of extracellular exosomes |
| WO2022042704A1 (en) * | 2020-08-28 | 2022-03-03 | 广东乾晖生物科技有限公司 | Decellularized matrix hydrogel, preparation method therefor, and application thereof |
| CN114159470A (en) * | 2021-12-10 | 2022-03-11 | 陈刚 | Preparation method of Eze-ECM hydrogel |
| CN114848833A (en) * | 2022-05-13 | 2022-08-05 | 中山大学中山眼科中心 | Method for assisting cell delivery of torsos function by acellular matrix glue |
| CN114908034A (en) * | 2022-04-27 | 2022-08-16 | 创芯国际生物科技(广州)有限公司 | Acellular fluid, acellular matrix hydrogel and preparation method and application thereof |
| CN115245593A (en) * | 2021-12-09 | 2022-10-28 | 上海市第六人民医院 | Dermal acellular extracellular matrix hydrogel and preparation method and application thereof |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114984319B (en) * | 2022-05-16 | 2023-12-12 | 上海市同济医院 | Preparation method of double-network hydrogel composite bracket capable of being printed in 3D (three-dimensional) |
| CN114870091B (en) * | 2022-05-17 | 2023-07-21 | 哈尔滨医科大学 | Bio-friendly myocardial dressing and preparation method and application thereof |
| CN115006600A (en) * | 2022-07-04 | 2022-09-06 | 杭州倍荣生物科技有限公司 | Preparation method and application of temperature-sensitive cornea repair hydrogel with biological activity |
| CN115287181A (en) * | 2022-09-28 | 2022-11-04 | 北京大学第三医院（北京大学第三临床医学院） | Procedural tissue graft decellularization device and method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103656749A (en) * | 2013-12-10 | 2014-03-26 | 北华大学 | Composite degradable antibacterial artificial cerebral dura mater and preparation method thereof |
| US20180256642A1 (en) * | 2013-08-02 | 2018-09-13 | The Trustees Of Columbia University In The City Of New York | Biomaterials derived from tissue extracellular matrix |
| CN111265719A (en) * | 2014-03-21 | 2020-06-12 | 匹兹堡大学-联邦高等教育体系 | Method for preparing final sterilized hydrogel from extracellular matrix |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BRPI0822802A2 (en) * | 2008-10-10 | 2015-09-01 | Histocell Sl | Biomaterial in the form of injectable hydrogel, biomaterial derived from the human umbilical cord, process for obtaining biomaterial, process for obtaining biomaterial obtained and use of biomaterial |
| CN105169483B (en) * | 2015-10-20 | 2018-10-12 | 中山大学 | A kind of preparation method and its acellular matrix gel of acellular matrix gel |
| CN111450119A (en) * | 2019-01-21 | 2020-07-28 | 南开大学 | Perinatal tissue-derived extracellular matrix hydrogel preparation for promoting organ injury repair |
| CN110975010B (en) * | 2019-11-25 | 2022-02-15 | 银丰低温医学科技有限公司 | Placenta tissue matrix material and preparation method thereof |
| CN112089890A (en) * | 2020-08-28 | 2020-12-18 | 广东乾晖生物科技有限公司 | Acellular matrix hydrogel and preparation method and application thereof |
-
2020
- 2020-08-28 CN CN202010889600.3A patent/CN112089890A/en active Pending
-
2021
- 2021-08-27 WO PCT/CN2021/115121 patent/WO2022042704A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20180256642A1 (en) * | 2013-08-02 | 2018-09-13 | The Trustees Of Columbia University In The City Of New York | Biomaterials derived from tissue extracellular matrix |
| CN103656749A (en) * | 2013-12-10 | 2014-03-26 | 北华大学 | Composite degradable antibacterial artificial cerebral dura mater and preparation method thereof |
| CN111265719A (en) * | 2014-03-21 | 2020-06-12 | 匹兹堡大学-联邦高等教育体系 | Method for preparing final sterilized hydrogel from extracellular matrix |
Non-Patent Citations (2)
| Title |
|---|
| 曹玉伦等: ""肝脏细胞外基质水凝胶的制备及表征"", 《中国组织工程研究》 * |
| 杨涛等主编: "《基因诊断与细胞治疗》", 31 August 2018, 科学技术文献出版社 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022042704A1 (en) * | 2020-08-28 | 2022-03-03 | 广东乾晖生物科技有限公司 | Decellularized matrix hydrogel, preparation method therefor, and application thereof |
| CN113082295A (en) * | 2021-04-02 | 2021-07-09 | 大连理工大学 | Derived scaffold based on skin-derived acellular matrix and construction method thereof |
| CN113881627A (en) * | 2021-10-21 | 2022-01-04 | 焕生汇生物基因技术(北京)有限公司 | Method for improving yield of extracellular exosomes |
| CN115245593A (en) * | 2021-12-09 | 2022-10-28 | 上海市第六人民医院 | Dermal acellular extracellular matrix hydrogel and preparation method and application thereof |
| CN114159470A (en) * | 2021-12-10 | 2022-03-11 | 陈刚 | Preparation method of Eze-ECM hydrogel |
| CN114908034A (en) * | 2022-04-27 | 2022-08-16 | 创芯国际生物科技(广州)有限公司 | Acellular fluid, acellular matrix hydrogel and preparation method and application thereof |
| CN114848833A (en) * | 2022-05-13 | 2022-08-05 | 中山大学中山眼科中心 | Method for assisting cell delivery of torsos function by acellular matrix glue |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2022042704A1 (en) | 2022-03-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112089890A (en) | Acellular matrix hydrogel and preparation method and application thereof | |
| US20220211755A1 (en) | Biomaterials derived from tissue extracellular matrix | |
| CN110975010B (en) | Placenta tissue matrix material and preparation method thereof | |
| CN105188787A (en) | Method for decellularization of tissue grafts | |
| CN102933705A (en) | Decellularized adipose tissue | |
| KR20100041027A (en) | Method for preparing three-dimensional scaffold using animal tissue powder and three-dimensional scaffold manufactured by the same method | |
| US20220280694A1 (en) | Detergent-free decellularized extracellular matrix preparation methods and bioinks for 3d printing | |
| CN105288737A (en) | Tissue engineering cartilage composite scaffold and preparation method thereof | |
| US20040187877A1 (en) | Method for repair of liver tissue | |
| CN113846050A (en) | Preparation method of tissue organoid | |
| Sahranavard et al. | Three-dimensional bio-printing of decellularized extracellular matrix-based bio-inks for cartilage regeneration: a systematic review | |
| CN104383601A (en) | Skeletal muscle acellular matrix biological patch and preparation method thereof | |
| Song et al. | Development of novel apoptosis-assisted lung tissue decellularization methods | |
| CN102861358B (en) | A kind of with umbilical cord jelly of Wharton for material makes new bio cartilage frame | |
| CN108245708A (en) | A kind of induction regenerated bioactive bracket of tendon tissue and its preparation method and application | |
| Akbarzadeh et al. | Decellularised whole ovine testis as a potential bio-scaffold for tissue engineering | |
| JP2009538854A (en) | Isolated natural natural collagen | |
| Peirsman et al. | Vascularized adipose tissue engineering: moving towards soft tissue reconstruction | |
| Sesli et al. | Decellularization of rat adipose tissue, diaphragm, and heart: a comparison of two decellularization methods | |
| US20150344842A1 (en) | Method for production of decellularized biological material and the decellularized biological material prepared therefrom | |
| CN104771784B (en) | One kind tissue de-cell liquid | |
| CN114686421B (en) | Preparation method and application of lung tissue extracellular matrix-free microcarrier | |
| CN113278587B (en) | Three-dimensional engineered breast cancer lung metastasis model, construction method and application | |
| WO2015168254A1 (en) | Method of preparing artificial organs and related compositions | |
| CN115216441A (en) | Composite scaffold for three-dimensional culture of stem cells and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201218 |