CN114191451A - Method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells - Google Patents
Method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells Download PDFInfo
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- CN114191451A CN114191451A CN202111545939.2A CN202111545939A CN114191451A CN 114191451 A CN114191451 A CN 114191451A CN 202111545939 A CN202111545939 A CN 202111545939A CN 114191451 A CN114191451 A CN 114191451A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
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- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
Abstract
The invention discloses a method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells, which comprises the following steps: preparing mesenchymal stem cells, preparing mesenchymal stem cell anti-inflammatory factors, combining the mesenchymal stem cell anti-inflammatory factors and the mesenchymal stem cells for local injection, carrying out venous return on the mesenchymal stem cells, and regularly following each index change of a user. The invention has the beneficial effects that: according to the method for delaying the chronic bone injury by adopting the umbilical cord mesenchymal stem cells, local injection is carried out by combining the mesenchymal stem cell anti-inflammatory factors and the mesenchymal stem cells, and the mesenchymal stem cells are returned to the vein, so that a good effect is obtained in the chronic bone injury, and the curative effect is remarkable.
Description
Technical Field
The invention relates to a method for delaying chronic bone injury, in particular to a method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells, and belongs to the technical field of medical treatment.
Background
Chronic bone injury is typically caused by long-term friction local to the joint, an inflammatory disease caused by inflammation, infection, degeneration, trauma, and other factors. Clinically manifested as redness, swelling, heat, pain, dysfunction and bone deformity. Serious people cause disability and affect the quality of life. The current treatment methods are: firstly, the patient needs to reduce the activity, avoids the stress of injured bone tissues, can select the medicine for promoting blood circulation and removing blood stasis for treatment, and adopts the traditional Chinese medicine massage, physical therapy, acupuncture and moxibustion, external medicine or matched anti-inflammatory and analgesic medicine for treatment. Conventional treatments do not control the continued occurrence of the injury.
The stem cell industry has become an emerging health medicine industry as part of regenerative medicine. Researchers find that mesenchymal stem cells are cells with self-renewal and multi-differentiation capabilities, and have easily obtained materials and wide sources. Can be differentiated into various tissues such as bone, cartilage, fat and the like, and has the function of immunoregulation. After the mesenchymal stem cells are broken, a large amount of anti-inflammatory factors can be generated. The application of mesenchymal stem cells in chronic injury of bones is a new treatment mode.
Disclosure of Invention
The invention aims to provide a method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells, so as to solve the problems in the background technology.
The invention realizes the aim through the following technical scheme, and a method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells comprises the following steps:
the method comprises the following steps: preparing mesenchymal stem cells and preparing mesenchymal stem cell anti-inflammatory factors;
step two: combining the mesenchymal stem cell anti-inflammatory factor with the mesenchymal stem cell for local injection;
performing intravenous infusion of the mesenchymal stem cells;
step three: and regularly visiting each index change of the user.
Preferably, in the step one, the mesenchymal stem cell is prepared by: and (3) taking the prepared P3 generation seed cells out of the liquid nitrogen, inoculating the seed cells into a serum-free culture medium for subculture, and carrying out culture in a culture repair box to mark P4 generation cells.
Preferably, the mesenchymal stem cell fusion degree is 90% or more, the mesenchymal stem cells are collected, and a part of the collected mesenchymal stem cells is frozen using 2 x 10 per one cell7Freezing and storing in a refrigerator at the temperature of-80 ℃; part of the mesenchymal stem cells were administered with saline at 2 x 10 per branch7The/ml was frozen in a liquid nitrogen tank and then waited for aseptic, viability, flow results.
Preferably, the anti-inflammatory factor of the mesenchymal stem cells is obtained by extracting the mesenchymal stem cells in a physical freezing and crushing mode, taking out the mesenchymal stem cells frozen by saline water, recovering the mesenchymal stem cells in a water bath kettle at 37 ℃, putting the mesenchymal stem cells into liquid nitrogen again for 10min after the mesenchymal stem cells are completely dissolved, and recovering the mesenchymal stem cells in the water bath kettle again after the mesenchymal stem cells are taken out; centrifuging the recovered cell suspension at 2000r for 10min to remove cell debris, and leaving the rest as mesenchymal stem cell inflammatory factors;
the preparation method of the mesenchymal stem cells comprises the following steps: and (3) taking the mesenchymal stem cells frozen in the frozen stock solution, completing recovery in a water bath kettle at 37 ℃ for 1min, wherein the rotating speed of the first washing is 1000r, the duration is 10min, the aperture of a filter screen is 70um, the filter screen filters out dead cells, and the rotating speed of the second washing is 1300r and lasts for 6 min.
Preferably, in step two, 2ml of the anti-inflammatory factor of the mesenchymal stem cells and 2 x 10 of the mesenchymal stem cells are added7As 1 part of preparation, the injection parts are selected according to the clinical condition of local injury, and the preparation is matched with the mesenchymal stem cells for venous return, and the preparation of the mesenchymal stem cells is as follows: mesenchymal stem cells 5 x 10795ml of physiological saline and 5ml of albumin are mixed, and the mixture is locally injected for 1 time and is matched with intravenous infusion for 1 time, 3 times are taken as a treatment course, and the interval of each time is 1 month.
Preferably, the site of local injection is kept clean to avoid infection, and a light diet should be taken within two weeks to avoid taking allergic food.
The invention has the beneficial effects that: the anti-inflammatory factor of the mesenchymal stem cells and the mesenchymal stem cells are combined for local injection and the mesenchymal stem cells are subjected to venous return, so that the anti-inflammatory agent has a good effect in chronic bone injury and an obvious curative effect.
Drawings
FIG. 1 is a schematic structural diagram of a preparation method 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.
Referring to fig. 1, a method for delaying chronic bone injury using umbilical cord mesenchymal stem cells, the first step is preparing mesenchymal stem cells and preparing anti-inflammatory factors of the mesenchymal stem cells.
And step two, (1) the anti-inflammatory factor of the mesenchymal stem cells and the mesenchymal stem cells are combined for local injection, and (2) the mesenchymal stem cells are returned back to the vein.
Third, precautions after local injection and intravenous return.
And fourthly, periodically follow-up the change of each index of the user.
The preparation process of the mesenchymal stem cells comprises the following steps: taking out the prepared P3 generation seed cells from liquid nitrogen, inoculating to serum-free culture medium for subculture, and culturing in a culture box to mark P4 generation cells. And (3) collecting the mesenchymal stem cells, wherein the fusion degree of the mesenchymal stem cells is more than 90%. Using freezing stock solution, 2 x 10 each for partial collected mesenchymal stem cells7Freezing and storing in a refrigerator at the temperature of-80 ℃; part of the mesenchymal stem cells were administered with saline at 2 x 10 per branch7The/ml is frozen in a liquid nitrogen tank. Wait for sterile, liveness, streaming results.
The anti-inflammatory factor of the mesenchymal stem cells is obtained by extracting the mesenchymal stem cells in a physical freezing and crushing mode, taking out the mesenchymal stem cells frozen in medium saline, recovering the mesenchymal stem cells in a water bath kettle at 37 ℃, putting the mesenchymal stem cells in liquid nitrogen again for 10min after the mesenchymal stem cells are completely dissolved, and recovering the mesenchymal stem cells in the water bath kettle again after the mesenchymal stem cells are taken out. Centrifuging the recovered cell suspension at 2000r for 10min to remove cell debris, and leaving the rest as the inflammatory factor of the mesenchymal stem cells.
The preparation method of the mesenchymal stem cells comprises the following steps: taking out the mesenchymal stem cells frozen in the frozen stock solution, recovering in a water bath kettle at 37 ℃ for 1min, washing for the first time at 1000r, 10min and 70um, filtering out dead cells by using a filter screen, and washing for the second time at 1300r and 6 min.
In the second step, (1) the anti-inflammatory factor of the mesenchymal stem cells and the mesenchymal stem cells are combined for local injection, and (2) the mesenchymal stem cells are returned to the vein.
Mixing 2ml of anti-inflammatory factor of mesenchymal stem cell and 2 x 10 of mesenchymal stem cell7As 1 part of the preparation, the number of parts to be injected is selected according to the clinical condition of the local lesion. Simultaneously, the preparation is matched with the mesenchymal stem cells for venous return, and the preparation of the mesenchymal stem cells is as follows: mesenchymal stem cells 5 x 10795ml of physiological saline and 5ml of albumin. The topical injection is administered for 1 time in combination with intravenous infusion for 1 time, and 3 times are a course of treatment, with 1 month interval.
The part of the local injection should be kept clean to avoid infection, a light diet should be paid attention to within two weeks to avoid eating allergic food, slight local red swelling and body temperature rise of the injection may occur after treatment, meanwhile, the phenomenon of sluggish or fatigue may also occur, smoking is reduced or stopped as much as possible, the spirit is not drunk at least within one month, the patient does not participate in strenuous exercise within 10 days, and the patient is not suitable for sauna, sweat steaming and the like within 7 days. Meanwhile, the method keeps good life style and good mood, enables the cells to play the best role and maintains for a longer time.
The whole using process is the result of performing knee joint cartilage defect treatment on the user for 6 to 24 months.
Clinical conditions:
1. selecting a crowd: the bone injury is judged clinically.
2. The using method comprises the following steps: (1) combining the anti-inflammatory factor of the mesenchymal stem cells with the mesenchymal stem cells for local injection, and (2) carrying out venous return on the mesenchymal stem cells.
3. The treatment results are as follows: the investigation is carried out according to the age and clinical condition of the selected population, and the follow-up time is 6-24 months.
4. Effective in cartilage regeneration, inflammation, redness and swelling, and pain disappearance or alleviation.
And (4) conclusion: according to the using condition results, the local injection is carried out by combining the mesenchymal stem cell anti-inflammatory factor and the mesenchymal stem cell, and the mesenchymal stem cell is subjected to venous return, so that the good effect is obtained in chronic bone injury, and the curative effect is obvious.
The specific implementation case is as follows: bone density was tested after 1 course of reinfusion of 39 persons using cord blood stem cells in combination with cord blood plasma, of which 18 persons all had mild symptoms of myocardial ischemia.
The table is a statistical table of the change in ventricular ejection fraction after 18 persons use:
as can be seen from the data in the table, there was a significant improvement in ventricular ejection fraction in all 18 mild patients. The symptoms of palpitation, chest distress and short breath are obviously improved.
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 (6)
1. A method for delaying chronic bone injury by adopting umbilical cord mesenchymal stem cells is characterized by comprising the following steps:
the method comprises the following steps: preparing mesenchymal stem cells and preparing mesenchymal stem cell anti-inflammatory factors;
step two: combining the mesenchymal stem cell anti-inflammatory factor with the mesenchymal stem cell for local injection;
performing intravenous infusion of the mesenchymal stem cells;
step three: and regularly visiting each index change of the user.
2. The method for delaying chronic bone injury by using umbilical cord mesenchymal stem cells according to claim 1, wherein the method comprises the following steps: in the first step, the mesenchymal stem cell is prepared by the following steps: and (3) taking the prepared P3 generation seed cells out of the liquid nitrogen, inoculating the seed cells into a serum-free culture medium for subculture, and carrying out culture in a culture repair box to mark P4 generation cells.
3. The method for delaying chronic bone injury by using umbilical cord mesenchymal stem cells according to claim 2, wherein the method comprises the following steps: collecting mesenchymal stem cells with a fusion degree of 90% or more, and freezing the collected mesenchymal stem cells to 2 × 10 cells per cell7Freezing and storing in a refrigerator at the temperature of-80 ℃; part of the mesenchymal stem cells were administered with saline at 2 x 10 per branch7The/ml was frozen in a liquid nitrogen tank and then waited for aseptic, viability, flow results.
4. The method for delaying chronic bone injury by using umbilical cord mesenchymal stem cells according to claim 3, wherein the method comprises the following steps:
the anti-inflammatory factor of the mesenchymal stem cells is obtained by extracting the mesenchymal stem cells in a physical freezing and crushing mode, taking out the mesenchymal stem cells frozen by saline water, recovering the mesenchymal stem cells in a water bath kettle at 37 ℃, putting the mesenchymal stem cells in liquid nitrogen again for 10min after the mesenchymal stem cells are completely dissolved, and recovering the mesenchymal stem cells in the water bath kettle again; centrifuging the recovered cell suspension at 2000r for 10min to remove cell debris, and leaving the rest as mesenchymal stem cell inflammatory factors;
the preparation method of the mesenchymal stem cells comprises the following steps: and (3) taking the mesenchymal stem cells frozen in the frozen stock solution, completing recovery in a water bath kettle at 37 ℃ for 1min, wherein the rotating speed of the first washing is 1000r, the duration is 10min, the aperture of a filter screen is 70um, the filter screen filters out dead cells, and the rotating speed of the second washing is 1300r and lasts for 6 min.
5. The method for delaying chronic bone injury by using umbilical cord mesenchymal stem cells according to claim 1, wherein the method comprises the following steps:
in the second step, 2ml of anti-inflammatory factor of mesenchymal stem cells and 2 x 10 mesenchymal stem cells are added7As 1 part of preparation, the injection parts are selected according to the clinical condition of local injury, and the preparation is matched with the mesenchymal stem cells for venous return, and the preparation of the mesenchymal stem cells is as follows: mesenchymal stem cells 5 x 10795ml of physiological saline and 5ml of albumin are mixed, and the mixture is locally injected for 1 time and is matched with intravenous infusion for 1 time, 3 times are taken as a treatment course, and the interval of each time is 1 month.
6. The method for delaying chronic bone injury by using umbilical cord mesenchymal stem cells according to claim 1, wherein the method comprises the following steps: the part for local injection is kept clean to avoid infection, and light diet is needed within two weeks to avoid taking allergic food.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108865986A (en) * | 2018-06-29 | 2018-11-23 | 马琳 | For repairing articular cartilage damage/defect mescenchymal stem cell preparation and its preparation method and application |
| CN109820869A (en) * | 2019-03-09 | 2019-05-31 | 和携科技(北京)有限公司 | A kind of preparation method of source of people umbilical cord mesenchymal stem cells injection that injecting treatment osteoarthritis for lacuna |
| CN110339212A (en) * | 2019-08-02 | 2019-10-18 | 陕西佰傲干细胞再生医学有限公司 | Mescenchymal stem cell preparation and its preparation method and application |
| CN112190596A (en) * | 2020-09-14 | 2021-01-08 | 陕西佰傲干细胞再生医学有限公司 | Mesenchymal stem cell preparation for treating arthritis and preparation method thereof |
| CN112294845A (en) * | 2020-09-21 | 2021-02-02 | 国大生命科学产业集团(深圳)有限公司 | Synovial mesenchymal stem cell combined PRP preparation for repairing articular cartilage damage and preparation method and application thereof |
-
2021
- 2021-12-16 CN CN202111545939.2A patent/CN114191451A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108865986A (en) * | 2018-06-29 | 2018-11-23 | 马琳 | For repairing articular cartilage damage/defect mescenchymal stem cell preparation and its preparation method and application |
| CN109820869A (en) * | 2019-03-09 | 2019-05-31 | 和携科技(北京)有限公司 | A kind of preparation method of source of people umbilical cord mesenchymal stem cells injection that injecting treatment osteoarthritis for lacuna |
| CN110339212A (en) * | 2019-08-02 | 2019-10-18 | 陕西佰傲干细胞再生医学有限公司 | Mescenchymal stem cell preparation and its preparation method and application |
| CN112190596A (en) * | 2020-09-14 | 2021-01-08 | 陕西佰傲干细胞再生医学有限公司 | Mesenchymal stem cell preparation for treating arthritis and preparation method thereof |
| CN112294845A (en) * | 2020-09-21 | 2021-02-02 | 国大生命科学产业集团(深圳)有限公司 | Synovial mesenchymal stem cell combined PRP preparation for repairing articular cartilage damage and preparation method and application thereof |
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