CN113975468A - Periosteum tissue aging repair agent and preparation process thereof - Google Patents
Periosteum tissue aging repair agent and preparation process thereof Download PDFInfo
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61L27/38—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 containing added animal cells
- A61L27/3804—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 containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3821—Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells
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- 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
- A61L27/38—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 containing added animal cells
- A61L27/3804—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 containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
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- 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
- A61L27/38—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 containing added animal cells
- A61L27/3839—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 containing added animal cells characterised by the site of application in the body
- A61L27/3843—Connective tissue
- A61L27/3847—Bones
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- 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
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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Abstract
The invention discloses an aging repair agent for periosteum tissues and a preparation process thereof, and the key points of the technical scheme are as follows: the periosteum tissue aging repair agent comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 1-3 parts of sodium hyaluronate, 40-50 parts of physiological saline, 0.2-0.5 part of osteogenesis inducer, 2-4 parts of fibrin and 1-3 parts of amino acid; also comprises umbilical cord stem cells occupying (6-7) multiplied by 100/mL in the biological preparation. The periosteum tissue aging repair agent has simple components, except necessary lubricating components, such as; the sodium hyaluronate, the normal saline, the osteogenesis inducer, the fibrin and the amino acid which form the biological preparation also have umbilical cord stem cells which can divide and differentiate, and the repairing effect is good.
Description
Technical Field
The invention relates to the field of orthopedic repair, in particular to an aging repair agent for periosteal tissues and a preparation process thereof.
Background
Periosteum, which is a dense connective tissue membrane covering almost all bone surfaces except joints, is composed of an outer fibrous layer and an inner cambium layer, and plays an important role in bone development and fracture healing. The fibrous layer is rich in vascular network, nerves, fiber cells, collagen and elastic fibers, so that the skeleton has activity, the mechanical stability of the periosteum is provided, and the key point of osteogenesis is; the inner cambium stores a large number of different types of cells, which play an important role in bone growth and repair.
The prior Chinese patent with publication number CN107812234A discloses a periosteum material with a tissue oxygenation function and a preparation method and application thereof, and comprises preparation of PLGA-coated PFOB particles (PLGA-PFOB), preparation of electrostatic spinning solution and preparation of the periosteum material. All the adopted raw materials are approved medical materials, and the medical materials have specific functions and high safety through special process steps; the periosteum material prepared by the invention has an oxygenation function, can solve the problem that the artificial periosteum is lack of blood supply in the initial stage of transplantation relative to the autologous periosteum, provides necessary oxygen for cell proliferation and accelerates the speed of tissue healing; the periosteum prepared by the electrostatic spinning method has a proper pore structure, is beneficial to bone cell growth and bone tissue repair, and is simple in process and easy to transform.
The above-mentioned patent has some disadvantages, such as: the cost is expensive to obtain, and the preparation process is complex.
Disclosure of Invention
In view of the problems mentioned in the background art, it is an object of the present invention to provide an agent for repairing aging of a membranous tissue and a process for preparing the same, which can solve the problems mentioned in the background art.
The technical purpose of the invention is realized by the following technical scheme:
the aging repair agent for the periosteal tissue comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 1-3 parts of sodium hyaluronate, 40-50 parts of physiological saline, 0.2-0.5 part of osteogenesis inducer, 2-4 parts of fibrin and 1-3 parts of amino acid; also comprises umbilical cord stem cells occupying (6-7) multiplied by 100/mL in the biological preparation.
Preferably, the biological agent comprises the following components in percentage by mass: 2 parts of sodium hyaluronate, 45 parts of physiological saline, 0.4 part of osteogenesis inducer, 3 parts of fibrin and 2 parts of amino acid; also included are umbilical cord stem cells occupying 6.6 x 100/mL in the biological preparation.
Preferably, the umbilical cord stem cells are P2-P5 generation umbilical cord stem cells.
Preferably, the biological preparation also comprises osteoblasts occupying 1X 100/mL of the biological preparation.
The invention also provides a preparation process of the periosteum tissue aging repair agent, which is characterized by comprising the following steps: the method comprises the following steps:
s1, culturing umbilical cord stem cells: subculturing the umbilical cord stem cells to obtain P2-P5 generation umbilical cord stem cells, and collecting culture supernatant of the P2-P5 generation umbilical cord stem cells to obtain an umbilical cord stem cell conditioned medium;
s2, osteoblast culture: subculturing osteoblasts to obtain P2-P3 generation osteoblasts, and collecting culture supernatant of the P2-P3 generation osteoblasts to obtain an osteoblast conditioned medium;
s3, primary mixing: mixing sodium hyaluronate, normal saline and osteogenesis inducer at 1-5 ℃ to obtain a first mixed solution;
s4, secondary mixing: adding fibrin and amino acid into the primary mixed solution at 4-8 deg.C to obtain a second mixed solution;
s5, mixing for three times: separating the umbilical cord stem cells in the umbilical cord stem cell conditioned medium from the osteoblast cells in the osteoblast conditioned medium by using a cell separator, and adding the cells together with the suspension to the second mixture.
Preferably, when the S1 umbilical cord stem cells are cultured, the umbilical cord stem cells are subcultured to obtain P3 generation umbilical cord stem cells, and culture supernatant of the P3 generation umbilical cord stem cells is collected to obtain the umbilical cord stem cell conditioned medium.
Preferably, in the step of culturing the osteoblasts of S2, the osteoblasts are subcultured to obtain osteoblasts of P2 generation, and culture supernatant of the osteoblasts of P2 generation is collected to obtain an osteoblast conditioned medium.
Preferably, in the step of S3, sodium hyaluronate, physiological saline and osteogenesis inducer are mixed at 2 ℃ to obtain a first mixed solution.
In summary, the invention mainly has the following beneficial effects:
firstly, the ingredients of the periosteum tissue aging repair agent are simple, except that the agent contains necessary lubricating ingredients, such as; the sodium hyaluronate, the normal saline, the osteogenesis inducer, the fibrin and the amino acid which form the biological preparation also have umbilical cord stem cells which can divide and differentiate, and the repairing effect is good;
secondly, the preparation process of the periosteum tissue aging repair agent is simple, the production steps are few, the cost is low, and the applicability is strong.
Drawings
FIG. 1 is a block flow diagram of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The aging repair agent for the periosteal tissue comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 1 part of sodium hyaluronate, 40 parts of normal saline, 0.2 part of osteogenesis inducer, 2 parts of fibrin and 1 part of amino acid; also included are umbilical cord stem cells occupying 6 x 100/mL in the biological preparation.
Wherein, the umbilical cord stem cell is P2 generation umbilical cord stem cell.
Wherein, the preparation also comprises osteoblasts occupying 1 x 100/mL in the biological preparation.
Wherein, the ingredients of the periosteum tissue aging repair agent are simple, except that the agent contains necessary lubricating ingredients, such as; the sodium hyaluronate, the normal saline, the osteogenesis inducer, the fibrin and the amino acid which form the biological preparation also have umbilical cord stem cells which can divide and differentiate, and the repairing effect is good.
Example 2
The aging repair agent for the periosteal tissue comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 3 parts of sodium hyaluronate, 50 parts of normal saline, 0.5 part of osteogenesis inducer, 4 parts of fibrin and 3 parts of amino acid; also included are umbilical cord stem cells occupying 7 x 100/mL in the biological preparation.
Wherein, the umbilical cord stem cell is P5 generation umbilical cord stem cell.
Wherein, the preparation also comprises osteoblasts occupying 1 x 100/mL in the biological preparation.
Example 3
The aging repair agent for the periosteal tissue comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 2 parts of sodium hyaluronate, 45 parts of physiological saline, 0.4 part of osteogenesis inducer, 3 parts of fibrin and 2 parts of amino acid; also included are umbilical cord stem cells occupying 6.6 x 100/mL in the biological preparation.
Wherein, the umbilical cord stem cell is P4 generation umbilical cord stem cell.
Wherein, the preparation also comprises osteoblasts occupying 1 x 100/mL in the biological preparation.
Example 4
The aging repair agent for the periosteal tissue comprises a biological agent, wherein the biological agent comprises the following components in parts by mass: 1 part of sodium hyaluronate, 50 parts of normal saline, 0.2 part of osteogenesis inducer, 4 parts of fibrin and 1 part of amino acid; also included are umbilical cord stem cells occupying 6 x 100/mL in the biological preparation.
Wherein, the umbilical cord stem cell is P3 generation umbilical cord stem cell.
Wherein, the preparation also comprises osteoblasts occupying 1 x 100/mL in the biological preparation.
Example 5
Referring to fig. 1, this embodiment further provides a preparation process of the agent for repairing aging of periosteal tissue, including the following steps:
s1, culturing umbilical cord stem cells: subculturing the umbilical cord stem cells to obtain P2-P5 generation umbilical cord stem cells, and collecting culture supernatant of the P2-P5 generation umbilical cord stem cells to obtain an umbilical cord stem cell conditioned medium;
s2, osteoblast culture: subculturing osteoblasts to obtain P2-P3 generation osteoblasts, and collecting culture supernatant of the P2-P3 generation osteoblasts to obtain an osteoblast conditioned medium;
s3, primary mixing: mixing sodium hyaluronate, normal saline and osteogenesis inducer at 1-5 ℃ to obtain a first mixed solution;
s4, secondary mixing: adding fibrin and amino acid into the primary mixed solution at 4-8 deg.C to obtain a second mixed solution;
s5, mixing for three times: separating the umbilical cord stem cells in the umbilical cord stem cell conditioned medium from the osteoblast cells in the osteoblast conditioned medium by using a cell separator, and adding the cells together with the suspension to the second mixture.
When the S1 umbilical cord stem cells are cultured, subculturing the umbilical cord stem cells to obtain P3 generation umbilical cord stem cells, and collecting culture supernatant of the P3 generation umbilical cord stem cells to obtain an umbilical cord stem cell conditioned medium.
When the osteoblasts of S2 are cultured, subculturing the osteoblasts to obtain osteoblasts of P2 generation, and collecting culture supernatant of the osteoblasts of P2 generation to obtain an osteoblast conditioned medium.
Wherein, when the S3 is mixed for one time, the sodium hyaluronate, the physiological saline and the osteogenesis inducer are mixed at the temperature of 2 ℃ to obtain a first mixed solution.
Wherein, the ingredients of the periosteum tissue aging repair agent are simple, except that the agent contains necessary lubricating ingredients, such as; the sodium hyaluronate, the normal saline, the osteogenesis inducer, the fibrin and the amino acid which form the biological preparation also have umbilical cord stem cells which can divide and differentiate, and the repairing effect is good; the preparation process of the periosteum tissue aging repair agent is simple, the production steps are few, the cost is low, and the applicability is strong.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. An agent for repairing aging of a periosteal tissue, characterized in that: the biological agent comprises a biological agent, and the biological agent comprises the following components in parts by mass: 1-3 parts of sodium hyaluronate, 40-50 parts of physiological saline, 0.2-0.5 part of osteogenesis inducer, 2-4 parts of fibrin and 1-3 parts of amino acid; also comprises umbilical cord stem cells occupying (6-7) multiplied by 100/mL in the biological preparation.
2. The agent for repairing aged membranous tissue according to claim 1, wherein: the biological agent comprises a biological agent, and the biological agent comprises the following components in parts by mass: 2 parts of sodium hyaluronate, 45 parts of physiological saline, 0.4 part of osteogenesis inducer, 3 parts of fibrin and 2 parts of amino acid; also included are umbilical cord stem cells occupying 6.6 x 100/mL in the biological preparation.
3. The agent for repairing aged membranous tissue according to claim 1, wherein: the umbilical cord stem cells are P2-P5 generation umbilical cord stem cells.
4. The agent for repairing aged membranous tissue according to claim 1, wherein: also included are osteoblasts occupying 1X 100/mL of the biological agent.
5. A preparation process of the periosteum tissue aging repair agent is characterized in that: the method comprises the following steps:
s1, culturing umbilical cord stem cells: subculturing the umbilical cord stem cells to obtain P2-P5 generation umbilical cord stem cells, and collecting culture supernatant of the P2-P5 generation umbilical cord stem cells to obtain an umbilical cord stem cell conditioned medium;
s2, osteoblast culture: subculturing osteoblasts to obtain P2-P3 generation osteoblasts, and collecting culture supernatant of the P2-P3 generation osteoblasts to obtain an osteoblast conditioned medium;
s3, primary mixing: mixing sodium hyaluronate, normal saline and osteogenesis inducer at 1-5 ℃ to obtain a first mixed solution;
s4, secondary mixing: adding fibrin and amino acid into the primary mixed solution at 4-8 deg.C to obtain a second mixed solution;
s5, mixing for three times: separating the umbilical cord stem cells in the umbilical cord stem cell conditioned medium from the osteoblast cells in the osteoblast conditioned medium by using a cell separator, and adding the cells together with the suspension to the second mixture.
6. The process for preparing an agent for repairing aging of membranous tissue according to claim 1, wherein: when the S1 umbilical cord stem cells are cultured, subculturing the umbilical cord stem cells to obtain P3 generation umbilical cord stem cells, and collecting culture supernatant of the P3 generation umbilical cord stem cells to obtain an umbilical cord stem cell conditioned medium.
7. The process for preparing an agent for repairing aging of membranous tissue according to claim 1, wherein: and when the S2 osteoblast is cultured, subculturing the osteoblast to obtain a P2 generation osteoblast, and collecting a culture supernatant of the P2 generation osteoblast to obtain an osteoblast conditioned medium.
8. The process for preparing an agent for repairing aging of membranous tissue according to claim 1, wherein: and when the S3 is mixed for the first time, mixing the sodium hyaluronate, the physiological saline and the osteogenesis inducer at the temperature of 2 ℃ to obtain a first mixed solution.
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| CN101247814A (en) * | 2005-06-13 | 2008-08-20 | 世源世龙股份有限公司 | Osteoblast composition of semi-solidified mixed fibrin for bone fracture agglutination and its manufacturing method |
| US20090324669A1 (en) * | 2006-07-26 | 2009-12-31 | The Regents Of The University Of California | Osteogenic enhancer composition |
| CN103480040A (en) * | 2013-09-27 | 2014-01-01 | 中国人民解放军第三军医大学第一附属医院 | Bone matrix material containing various proteins secreted by umbilical cord mesenchymal stem cells and preparation method thereof |
| CN107648669A (en) * | 2017-10-24 | 2018-02-02 | 武汉大学 | The method for building study of vascularized tissue engineering bone film |
| CN110199987A (en) * | 2019-07-22 | 2019-09-06 | 深圳市未来细胞生命科技有限公司 | A kind of mesenchyme stem cell preserving fluid and its store method and application |
| CN112451482A (en) * | 2020-11-30 | 2021-03-09 | 深圳博雅感知药业有限公司 | Method of treating osteoarthritis |
| CN112546073A (en) * | 2020-12-21 | 2021-03-26 | 吉林省熙达生物科技有限公司 | Mesenchymal stem cell preparation for treating osteoarthritis and preparation method and application thereof |
| CN113018317A (en) * | 2021-02-03 | 2021-06-25 | 上海兰天生物医药科技有限公司 | Application of mesenchymal stem cells and sodium hyaluronate in treatment of arthritis |
-
2021
- 2021-11-11 CN CN202111335034.2A patent/CN113975468A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10236976A (en) * | 1997-02-26 | 1998-09-08 | Chemo Sero Therapeut Res Inst | Osteoplasty promoter |
| CN101247814A (en) * | 2005-06-13 | 2008-08-20 | 世源世龙股份有限公司 | Osteoblast composition of semi-solidified mixed fibrin for bone fracture agglutination and its manufacturing method |
| US20090324669A1 (en) * | 2006-07-26 | 2009-12-31 | The Regents Of The University Of California | Osteogenic enhancer composition |
| CN103480040A (en) * | 2013-09-27 | 2014-01-01 | 中国人民解放军第三军医大学第一附属医院 | Bone matrix material containing various proteins secreted by umbilical cord mesenchymal stem cells and preparation method thereof |
| CN107648669A (en) * | 2017-10-24 | 2018-02-02 | 武汉大学 | The method for building study of vascularized tissue engineering bone film |
| CN110199987A (en) * | 2019-07-22 | 2019-09-06 | 深圳市未来细胞生命科技有限公司 | A kind of mesenchyme stem cell preserving fluid and its store method and application |
| CN112451482A (en) * | 2020-11-30 | 2021-03-09 | 深圳博雅感知药业有限公司 | Method of treating osteoarthritis |
| CN112546073A (en) * | 2020-12-21 | 2021-03-26 | 吉林省熙达生物科技有限公司 | Mesenchymal stem cell preparation for treating osteoarthritis and preparation method and application thereof |
| CN113018317A (en) * | 2021-02-03 | 2021-06-25 | 上海兰天生物医药科技有限公司 | Application of mesenchymal stem cells and sodium hyaluronate in treatment of arthritis |
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Application publication date: 20220128 |