WO2023027674A2 - Production of hydrogels and bioscaffolds originated from decellularized brain/spinal meninx for regenerative and reconstructive medical applications - Google Patents
Production of hydrogels and bioscaffolds originated from decellularized brain/spinal meninx for regenerative and reconstructive medical applications Download PDFInfo
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- WO2023027674A2 WO2023027674A2 PCT/TR2022/050899 TR2022050899W WO2023027674A2 WO 2023027674 A2 WO2023027674 A2 WO 2023027674A2 TR 2022050899 W TR2022050899 W TR 2022050899W WO 2023027674 A2 WO2023027674 A2 WO 2023027674A2
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- bioscaffolds
- hydrogels
- decellularized
- production
- growth factor
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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
- 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
- A61L27/14—Macromolecular materials
- A61L27/26—Mixtures of macromolecular compounds
-
- 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
- A61L27/3633—Extracellular matrix [ECM]
-
- 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
-
- 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
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Definitions
- the invention relates to the production of hydrogels and bioscaffolds that originate from decellularized brain/spinal meninx for regenerative and reconstructive medical applications that can be used in humans and animals and that are obtained from healthy human and/or animal tissue.
- Matrigel® A mouse tumor matrix (Engelbreth-Holm- Swarm (EHS) mouse sarcoma cells) extract called Matrigel® provides copious amounts of basal membrane protein sources such as laminin, collagen IV, heparan sulfate, etc. Matrigel® has a structure similar to the original matrix and gelates at room temperature. Embryonic tissue explants, stem cells, and various cell types differentiate when cultured in Matrigel®. Matrigel® is used in angiogenesis, cell proliferation, spheroid formation, organoid formation from a single cell, etc. in various in vitro studies.
- EHS Engelbreth-Holm- Swarm
- Matrigel® due to its murine origins and potential tumorigenicity, Matrigel® cannot be used in humans.
- its content is not well defined, the role of its components in biological activities is unknown, it is expensive, its content varies based on the production method, and its storage time is short.
- the object of the present invention is to provide the production of hydrogels and bioscaffolds that originate from decellularized brain/spinal meninx that can be used in humans and animals and that are obtained from healthy human and/or animal tissue.
- Another object of the present invention is to provide the production of hydrogels and bioscaffolds that can be easily obtained, that are inexpensive, and that have a longer storage time.
- Figure 1 The view of the production diagram of the hydrogels and bioscaffolds of the invention.
- the subject of the invention is the production of decellularized hydrogels and bioscaffolds for regenerative and reconstructive medical applications; and comprises the steps of basically accelerating tissue regeneration, differentiating the stem/progenitor cells, triggering angiogenesis and obtaining extracellular matrix (ECM) hydrogels and bioscaffolds by treating the brain or spinal meninx tissue which are the support material for stems/cells with decellularization technique.
- ECM extracellular matrix
- the subject of the invention is the production of decellularized hydrogels and bioscaffolds obtained from the brain or spinal meninx and may comprise one of the steps of;
- ECM hydrogels and bioscaffolds by themselves and/or with natural polymers such as vegetable mucilage, cellulose, alginate, chitosan, fibrin, elastin, collagen (of vertebrate and/or invertebrate origin), gelatin, decellularized matrices,
- ECM hydrogels and bioscaffolds by themselves and/or with synthetic polymers such as polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic-co-lactic acid (PLGA), poly-e-caprolactone (PCL), etc.
- synthetic polymers such as polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic-co-lactic acid (PLGA), poly-e-caprolactone (PCL), etc.
- ECM hydrogels and bioscaffolds with growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), Brain-derived neurotrophic factor (BDNF), transforming growth factor a (TGF-a), transforming growth factor P (TGF- P), and vascular endothelial growth factor (VEGF), etc.
- growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), Brain-derived neurotrophic factor (BDNF), transforming growth factor a (TGF-a), transforming growth factor P (TGF- P), and vascular endothelial growth factor (VEGF), etc.
- growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), Brain-derived neurotrophic factor (BDNF), transforming growth factor a (TGF
- the brain/spinal meninx as the subject of the invention is composed of three membranes (dura mater, arachnoid and pia mater) responsible for the protection of the brain and the spine.
- the Dura mater is the outermost membrane that surrounds the other meninx.
- the middle layer is the arachnoid and the innermost layer is the pia mater.
- the role of the meninx is to protect the central nervous system together with the cerebrospinal fluid. Apart from its protective function, the meninx regulates the pathological and physiological events of the central nervous system. Meninx contributes to neuro-homeostasis by secreting many trophic factors in adults.
- the micro-architecture of the meninx contains a laminin-rich extracellular matrix that secretes growth factors that have the potential to modulate stem cell homeostasis and cortical function.
- the extracellular matrix of the meninx contains large amounts of collagen types I, III, and IV, as well as non-collagenous proteins such as tenascin and fibronectin.
- the extracellular matrix of the meninx contains basic fibroblast growth factor, insulin-like growth factor-II, stromal cell-derived factor I, retinoic acid and heparan sulfate.
- the said material can be obtained from slaughterhouses.
- the meninx (1) is first separated from the brain or the spinal cord obtained from the slaughterhouse or cadaver mechanically (2). After that, unwanted tissues such as fat, etc. are removed from the meninx (3) and irrigated thoroughly (4) to dispose of blood, etc. residues.
- the meninx is divided into approximately 2 centimetres (cm) x 2 cm pieces.
- the decellularization can be performed chemically or with supercritical carbon dioxide (CO2) /ethanol. In the said technique, the decellularization treatment is performed by means of chemical methods.
- the meninx is first treated for 24 hours with hypotonic 10 millimolar (mM) Trizma hydrochloride and 5 mM EDTA solution (pH 8.0) (5).
- the meninx is irrigated with ultra pure water for 24 hours (4) after the hypotonic solution.
- the materials are treated in a magnetic agitator for 48 hours with hypotonic 50 mM Trizma hydrochloride, 5 mM EDTA solution (pH 8.0) comprising 1% Triton X-100 (v/v) detergent (6).
- the meninx is washed 24 hours with ultra pure water (4) in order to remove the detergent residue.
- the materials are incubated at 37.5 °C for 24 hours with Deoxyribonuclease (DNase) (200 micrograms/milliliter (pg/mL)) and Ribonuclease (RNase) (50 pg/mL) solutions prepared in 10 mM magnesium chloride (MgCl) and 50 mM Trizma (pH: 7.5) buffer (7).
- DNase Deoxyribonuclease
- RNase Ribonuclease
- the scaffolds are left for agitation to be washed for 24 hours with pure water (4).
- the decellularized meninx which is purified from chemical residues by irrigation (9) is placed in a tube containing 20 mL of ultra pure water for 5 g of wet tissue and homogenized for 10 minutes with a cool pack to prevent the temperature from rising too much (10).
- the homogenized solution is molded (11) and left overnight at -26°C to be readied for freeze- drying. Prepared materials are freeze-dried for 24 hours (12).
- the freeze-dried meninges are taken from the petri dish and ground into a powder with the help of a grinder (13).
- Centrifugation is performed at 14,000 rpm for 5 minutes to remove undigested particles from the pre-gel (the gel that has not been neutralized as a result of digestion) (15).
- the pre-gel is drawn carefully with the help of a micropipette and taken into a vial.
- the vial is left at +4°C until it is used (the pre-gel rests at +4°C for at least 1 day before it is neutralized).
- the hydrogel and bioscaffold of the invention are obtained from healthy tissue and are also treated with many chemicals and enzymes during decellularization. In the final step, it is treated with the enzyme pepsin to obtain a gel. All of these steps minimize the nuclear materials (DNA and RNA) at the origin of the material that causes tissue rejection and/or host response. Therefore, the hydrogel and bioscaffold of the invention can be used in humans. Its content was determined by proteomic analysis. Since Matrigel® is of tumorigenic origin, its content varies from production to production. Since the invention is obtained from healthy tissue, its content does not change based on the production. It is cheaper because it can be obtained easily. The analyses revealed that the storage time of the invention is much longer than that of Matrigel®.
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Abstract
The invention relates to the production of hydrogels and bioscaffolds that originate from decellularized brain/ spinal meninx for regenerative and reconstructive medical applications that can be used in humans and that are obtained from healthy tissue.
Description
PRODUCTION OF HYDROGELS AND BIOSCAFFOLDS ORIGINATED FROM DECELLULARIZED BRAIN/SPINAL MENINX FOR REGENERATIVE AND RECONSTRUCTIVE MEDICAL APPLICATIONS
Technical Field
The invention relates to the production of hydrogels and bioscaffolds that originate from decellularized brain/spinal meninx for regenerative and reconstructive medical applications that can be used in humans and animals and that are obtained from healthy human and/or animal tissue.
Prior Art
A mouse tumor matrix (Engelbreth-Holm- Swarm (EHS) mouse sarcoma cells) extract called Matrigel® provides copious amounts of basal membrane protein sources such as laminin, collagen IV, heparan sulfate, etc. Matrigel® has a structure similar to the original matrix and gelates at room temperature. Embryonic tissue explants, stem cells, and various cell types differentiate when cultured in Matrigel®. Matrigel® is used in angiogenesis, cell proliferation, spheroid formation, organoid formation from a single cell, etc. in various in vitro studies.
However, due to its murine origins and potential tumorigenicity, Matrigel® cannot be used in humans. In addition, its content is not well defined, the role of its components in biological activities is unknown, it is expensive, its content varies based on the production method, and its storage time is short.
Various improvements have been made in the art for the fabrication of hydrogels and bioscaffolds.
In the patent document numbered KR20170003916A in the state of the art, methods for preparing sterilized, gelated, solubilized extracellular matrix (ECM) compositions beneficial as cell growth matrices are mentioned.
In the patent document numbered JP2019513125A in the state of the art, a composition comprising nanovesicles derived from isolated extracellular matrix and a pharmaceutically acceptable carrier is described. Methods that generate ECM-derived nanovesicles are also described.
When the hydrogels and bioscaffolds present in the art were examined, a need for developing a hydrogel and bioscaffold obtained by using brain/spinal meninx emerged.
The object of the Invention
The object of the present invention is to provide the production of hydrogels and bioscaffolds that originate from decellularized brain/spinal meninx that can be used in humans and animals and that are obtained from healthy human and/or animal tissue.
Another object of the present invention is to provide the production of hydrogels and bioscaffolds that can be easily obtained, that are inexpensive, and that have a longer storage time.
Detailed Description of the Invention
The hydrogel and bioscaffold realized to achieve the purposes of the present invention are shown in the attached figure.
In this figure;
Figure 1: The view of the production diagram of the hydrogels and bioscaffolds of the invention.
The steps in the figure have been numbered individually, and the corresponding descriptions are given below.
1. Brain or spinal meninx
2. Mechanical separation
3. Removing unwanted tissue such as fat, etc. from the meninx
4. Irrigation
5. Treating with hypotonic Trizma hydrochloride and Ethylenediamine tetraacetic acid (EDTA) solution
6. Treating in the agitator
7. Incubating at 37.5° C for 24 hours with the solutions
8. Decontamination
9. Obtaining decellularized meninx purged from chemical residues by means of irrigation
10. Homogenization of the decellularized meninx
11. Moulding of the homogenized solution
12. Freeze drying
13. Pulverizing with a grinder
14. Leaving for digestion for 72 hours in the magnetic stirrer
15. Centrifuging
16. Neutralizing
17. Incubating
18. MeninGEL Hydrogel
19. MeninRIX bioscaffold
The subject of the invention is the production of decellularized hydrogels and bioscaffolds for regenerative and reconstructive medical applications; and comprises the steps of basically accelerating tissue regeneration, differentiating the stem/progenitor cells, triggering angiogenesis and obtaining extracellular matrix (ECM) hydrogels and bioscaffolds by treating the brain or spinal meninx tissue which are the support material for stems/cells with decellularization technique.
The subject of the invention is the production of decellularized hydrogels and bioscaffolds obtained from the brain or spinal meninx and may comprise one of the steps of;
Combining the obtained ECM hydrogels and bioscaffolds by themselves and/or with natural polymers such as vegetable mucilage, cellulose, alginate, chitosan, fibrin, elastin, collagen (of vertebrate and/or invertebrate origin), gelatin, decellularized matrices,
Combining the obtained ECM hydrogels and bioscaffolds by themselves and/or with
synthetic polymers such as polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic-co-lactic acid (PLGA), poly-e-caprolactone (PCL), etc.,
Combining the obtained ECM hydrogels and bioscaffolds with growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), Brain-derived neurotrophic factor (BDNF), transforming growth factor a (TGF-a), transforming growth factor P (TGF- P), and vascular endothelial growth factor (VEGF), etc.,
Combining the obtained ECM hydrogels and bioscaffolds with exosomes obtained from stem/cell and/or bodily fluids of human and/or animal origin,
Combining the obtained ECM hydrogels and bioscaffolds with cytokines, chemokines and/or hormones.
The brain/spinal meninx as the subject of the invention is composed of three membranes (dura mater, arachnoid and pia mater) responsible for the protection of the brain and the spine. The Dura mater is the outermost membrane that surrounds the other meninx. The middle layer is the arachnoid and the innermost layer is the pia mater. The role of the meninx is to protect the central nervous system together with the cerebrospinal fluid. Apart from its protective function, the meninx regulates the pathological and physiological events of the central nervous system. Meninx contributes to neuro-homeostasis by secreting many trophic factors in adults. If the micro-architecture of the meninx is examined, it is found that it contains a laminin-rich extracellular matrix that secretes growth factors that have the potential to modulate stem cell homeostasis and cortical function. Moreover, the extracellular matrix of the meninx contains large amounts of collagen types I, III, and IV, as well as non-collagenous proteins such as tenascin and fibronectin. In addition, the extracellular matrix of the meninx contains basic fibroblast growth factor, insulin-like growth factor-II, stromal cell-derived factor I, retinoic acid and heparan sulfate.
The said material can be obtained from slaughterhouses. The meninx (1) is first separated from the brain or the spinal cord obtained from the slaughterhouse or cadaver mechanically (2). After that, unwanted tissues such as fat, etc. are removed from the meninx (3) and irrigated thoroughly (4) to dispose of blood, etc. residues. To make the decellularization easier, the meninx is divided into approximately 2 centimetres (cm) x 2 cm pieces. The decellularization can be performed chemically or with supercritical carbon dioxide (CO2) /ethanol. In the said technique, the decellularization treatment is performed by means of
chemical methods. The meninx is first treated for 24 hours with hypotonic 10 millimolar (mM) Trizma hydrochloride and 5 mM EDTA solution (pH 8.0) (5). The meninx is irrigated with ultra pure water for 24 hours (4) after the hypotonic solution. Then, the materials are treated in a magnetic agitator for 48 hours with hypotonic 50 mM Trizma hydrochloride, 5 mM EDTA solution (pH 8.0) comprising 1% Triton X-100 (v/v) detergent (6). In the remainder of this procedure, the meninx is washed 24 hours with ultra pure water (4) in order to remove the detergent residue. Then, the materials are incubated at 37.5 °C for 24 hours with Deoxyribonuclease (DNase) (200 micrograms/milliliter (pg/mL)) and Ribonuclease (RNase) (50 pg/mL) solutions prepared in 10 mM magnesium chloride (MgCl) and 50 mM Trizma (pH: 7.5) buffer (7). The meninx is irrigated with ultra pure water several times at the end of this period (4). After the irrigation, decontamination is performed within a solution containing 4% ethanol (v/) and 0.1% peracidic acid (v/v) for 5 hours (8). Following decontamination, the scaffolds are left for agitation to be washed for 24 hours with pure water (4). The decellularized meninx which is purified from chemical residues by irrigation (9) is placed in a tube containing 20 mL of ultra pure water for 5 g of wet tissue and homogenized for 10 minutes with a cool pack to prevent the temperature from rising too much (10). The homogenized solution is molded (11) and left overnight at -26°C to be readied for freeze- drying. Prepared materials are freeze-dried for 24 hours (12). The freeze-dried meninges are taken from the petri dish and ground into a powder with the help of a grinder (13). Then, 20mM acetic acid, 1 mg/ml pepsin solution is prepared and mixed at a low speed on a magnetic stirrer. Finally, 10 mg/ml of decellularized meninx are added to the solution and allowed to digest for 72 hours in a magnetic stirrer (14).
Centrifugation is performed at 14,000 rpm for 5 minutes to remove undigested particles from the pre-gel (the gel that has not been neutralized as a result of digestion) (15). The pre-gel is drawn carefully with the help of a micropipette and taken into a vial. The vial is left at +4°C until it is used (the pre-gel rests at +4°C for at least 1 day before it is neutralized).
For neutralization, 10 pl of 7.5% sodium bicarbonate (NaHCO3) is added to each 100 pl of pre-gel and neutralized with a micropipette at +4°C (on a cool pack) (16).
Neutralized gels are incubated at 37°C for 1 hour (17) to obtain MeninGEL Hydrogel (18).
If the MeninGEL hydrogel is freeze-dried for one night (12), the MeninRIX bioscaffold is
obtained (19).
Said hydrogel is called MeninGEL and the bioscaffold is called MeninRIX bioscaffold. The hydrogel and bioscaffold of the invention are obtained from healthy tissue and are also treated with many chemicals and enzymes during decellularization. In the final step, it is treated with the enzyme pepsin to obtain a gel. All of these steps minimize the nuclear materials (DNA and RNA) at the origin of the material that causes tissue rejection and/or host response. Therefore, the hydrogel and bioscaffold of the invention can be used in humans. Its content was determined by proteomic analysis. Since Matrigel® is of tumorigenic origin, its content varies from production to production. Since the invention is obtained from healthy tissue, its content does not change based on the production. It is cheaper because it can be obtained easily. The analyses revealed that the storage time of the invention is much longer than that of Matrigel®.
Claims
1. The production of decellularized hydrogels and bioscaffolds for regenerative and reconstructive medical applications; comprising the step of accelerating tissue regeneration, differentiating the stem/progenitor cells, triggering angiogenesis and obtaining extracellular matrix (ECM) hydrogels and bioscaffolds by treating the brain or spinal meninx tissue which are the support material for stems/ cells obtained from humans and/or animals with decellularization technique.
2. The production of decellularized hydrogels and bioscaffolds according to Claim 1, characterized in that the obtained ECM hydrogels and bioscaffolds are combined by themselves and/or with natural polymers, synthetic polymers, growth factors, exosomes, cytokines, chemokines, and/or hormones.
3. The production of decellularized hydrogels and bioscaffolds according to Claim 2, characterized in that the natural polymers are vegetable mucilage, cellulose, alginate, chitosan, fibrin, elastin, collagen (of vertebrate and/or invertebrate origin), gelatin, and decellularized matrices.
4. The production of decellularized hydrogels and bioscaffolds according to Claim 2, characterized in that the synthetic polymers are polylactic acid (PLA), polyglycolic acid (PGA), polyglycolic-co-lactic acid (PLGA), and poly-e-caprolactone (PCL).
5. The production of decellularized hydrogels and bioscaffolds according to Claim 2, characterized in that the growth factors are epidermal growth factor (EGF), nerve growth factor (NGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), Brain-derived neurotrophic factor (BDNF), transforming growth factor a (TGF-a), transforming growth factor P (TGF-P), and vascular endothelial growth factor (VEGF).
6. The production of decellularized hydrogels and bioscaffolds according to Claim 2, characterized in that the exosomes are exosomes obtained from stem/cell and/or bodily fluids of human and/or animal origin.
7
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| Application Number | Priority Date | Filing Date | Title |
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| TR2021/013582 | 2021-08-27 | ||
| TR2021/013582A TR2021013582A2 (en) | 2021-08-27 | 2021-08-27 | PRODUCTION OF DECELLULARIZED BRAIN/SPINAL CORD MENINGE-DERIVED HYDROGELS AND BIOSCALES FOR REGENERATIVE AND RECONSTRUCTIVE MEDICINE APPLICATIONS |
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| WO2023027674A2 true WO2023027674A2 (en) | 2023-03-02 |
| WO2023027674A3 WO2023027674A3 (en) | 2023-05-04 |
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| TR (1) | TR2021013582A2 (en) |
| WO (1) | WO2023027674A2 (en) |
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| TR2021013582A2 (en) | 2021-09-21 |
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