CN109550084A - A kind of guide tissue regeneration film, sticking patch and the preparation method of compound BMSCs - Google Patents
A kind of guide tissue regeneration film, sticking patch and the preparation method of compound BMSCs Download PDFInfo
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- CN109550084A CN109550084A CN201710886334.7A CN201710886334A CN109550084A CN 109550084 A CN109550084 A CN 109550084A CN 201710886334 A CN201710886334 A CN 201710886334A CN 109550084 A CN109550084 A CN 109550084A
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- 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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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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
- 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
- A61L27/3834—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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- 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
- A61L27/38—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
- A61L27/3882—Hollow organs, e.g. bladder, esophagus, urether, uterus
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- 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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/005—Ingredients of undetermined constitution or reaction products thereof
-
- 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
-
- 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
- A61L2430/22—Materials or treatment for tissue regeneration for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
Abstract
The invention belongs to gynemetrics's technical fields, more particularly to the guide tissue regeneration film of compound BMSCs a kind of, sticking patch and preparation method, the present invention is to prepare single cell suspension after digesting the BMSCs pancreatin in growth period, single cell suspension is taken to drop evenly on guide tissue regeneration film, it is put into incubator and cultivates up to compound mesenchymal stem cell guide tissue regeneration film, using BMSCs of the present invention as the active constituent for the treatment of endometrial impairment, it is convenient to obtain, secreting growth factor improves local microenvironment and immunological regulation, there is good biocompatibility, promote damage endometrium reparation, increase inner film thickness and vessel density.
Description
Technical field
The invention belongs to gynemetrics's technical fields, and in particular to a kind of guide tissue regeneration film of compound BMSCs, sticking patch and
Preparation method.
Background technique
Inner membrance functionality repairs obstacle, instead connective tissue proliferation reparation, uterus front and back when endometrium seriously damages
Fibrosis occurs for wall, cicatrization, is clinically mainly shown as Asherman's syndrom, amenorrhoea and uterine infertile etc., there is no at present
Imitate treatment method.
Before making the present invention, as follows for the treatment method of serious uterine inner film injury: clinically universal method is uterine cavity
Videoendoscopic surgery mechanicalness separation Asherman's syndrom, postoperative placement of intrauterine devices or adherence preventing material (mainly hyaluronic acid) etc., and
Promote inner membrance growth in postoperative estrogen of giving.Disadvantage: endometrium is seriously damaged, inner membrance scarring problem can not be solved, no
It is able to achieve inner membrance functionality reparation, and easily sends out adhesion again；In addition, employment fresh amnion covering wound after Asherman's syndrom separation
Disadvantage: face not can induce scar inner membrance own functionality reparation, and amnion there is also immune inflammation and induces the risks such as infection；
In addition, granulocyte colony stimulating factor is perfused in uterine cavity, it was reported that patient's inner film thickness that part inner membrance can be made thin increases, and lacks
Point: cannot retain granulocyte colony stimulating factor in part, and action time is short, and local efficacy is poor, is also unable to reach promotion scar
The effect of inner membrance growth；Mesenchymal stem cell (Bonemarrow-derived mesenchymal stem cells,
BMSCs) mesenchymal stem cell is uniformly injected in uterine cavity after the dilatation and curettage of the uterus, it is postoperative to give hormone replacement therapy,
It can increase patient uterine's inner film thickness, disadvantage: being difficult to affect the treatment in damage local positioning after BMSCs locally injecting.
To solve the above-mentioned problems, the present invention provides the guide tissue regeneration film preparation methods of compound BMSCs a kind of, benefit
Piece and preparation method thereof can promote endometrium smooth muscle reparation, increase inner film thickness and local vascular density.
Summary of the invention
It is an advantage of the invention to provide the guide tissue regeneration film preparation methods of compound BMSCs a kind of, will
Mesenchymal stem cell is compound with guide tissue regeneration film, for treating serious uterine inner film injury；
Yet another object of the invention is that providing the guide tissue regeneration film of compound BMSCs a kind of, uterus can be promoted
Intimal smooth muscle reparation increases inner film thickness and local vascular density；
It is still another object of the present invention to provide a kind of sticking patch of guide tissue regeneration film comprising compound BMSCs, energy
It is enough in treatment serious uterine inner film injury.
To solve the above-mentioned problems, the present invention provides a kind of preparation method of the guide tissue regeneration film of compound BMSCs,
The following steps are included:
The BMSCs pancreatin in growth period is digested, single cell suspension is prepared；
Single cell suspension is taken to drop evenly on guide tissue regeneration film, it is spare；
The guide tissue regeneration film that single cell suspension is added dropwise is put into incubator and is cultivated, the guide tissue regeneration of BMSCs is obtained
Film.
Preferably, the cell quantity of the single cell suspension is 2.5 × 106A/cm2。
Preferably, single cell suspension is taken to drop evenly on the rough surface of guide tissue regeneration film.
Preferably, the guide tissue regeneration film that single cell suspension is added dropwise is put into incubator and is cultivated, obtained compound
Specific step is as follows for the guide tissue regeneration film of BMSCs:
It is 37 DEG C that the guide tissue regeneration film that single cell suspension is added dropwise, which is put into temperature, and contains 5%CO2Incubator in, culture
1h obtains the guide tissue regeneration film of compound BMSCs.
Preferably, the guide tissue regeneration film of the compound BMSCs of preparation is for treating endometrial impairment.
A kind of guide tissue regeneration film of compound BMSCs, is prepared using above-mentioned preparation method, has irregular collagen
The rough surface surface of beam arrangement and cell covering layer, the cell covering layer are attached on rough surface surface, and between collagen beam
There is cell connection.
Preferably, for treating endometrial impairment.
Preferably, endometrial impairment refers to endometrium excessive damage, and inner membrance functionality is repaired obstacle, instead tied
Hyperblastosis reparation is formed, fibrosis, one of cicatrization occur for uterine wall.
A kind of sticking patch, the guide tissue regeneration film including compound BMSCs, the guide tissue regeneration film of the compound BMSCs
It is prepared using above-mentioned preparation method.
Preferably, the sticking patch is for treating endometrial impairment.
Beneficial effects of the present invention:
1, the guide tissue regeneration film preparation method of compound BMSCs provided by the invention, by mesenchymal stem cell with draw
It is compound to lead tissue regeneration membrane, for treating serious uterine inner film injury.
2, the guide tissue regeneration film of compound BMSCs provided by the invention, by mesenchymal stem cell and guidance group
It knits regeneration membrane to be combined with each other, guides tissue film to provide for mesenchymal stem cell and support site, filled between raising local bone marrow
Matter concentration of stem cells extends its action time, preferably treatment endometrial impairment.
3, the guide tissue regeneration film of compound BMSCs provided by the invention can promote endometrium smooth muscle reparation,
Increase inner film thickness and local vascular density.
4, the guide tissue regeneration film of compound BMSCs provided by the invention, the mesenchymal stem cell used can
It is adhered to guide tissue regeneration film surface, there is good biocompatibility between guide tissue regeneration film.
5, the sticking patch of the guide tissue regeneration film provided by the invention comprising compound BMSCs, is used to treat endometrium
Damage, good effect.
Detailed description of the invention
Fig. 1 is the appearance diagram of guide tissue regeneration film of the invention；
Fig. 2 is the scanning electron microscope (SEM) photograph of guide tissue regeneration film of the invention；
Fig. 3 is flow cytometry BMSCs surface marker measurement schematic diagram of the invention；
Fig. 4 is mesenchymal stem cell directional induction in vitro differentiated result schematic diagram of the invention；
Fig. 5 is the scanning electron microscope (SEM) photograph on the guide tissue regeneration film surface of compound BMSCs of the invention；
Fig. 6 is that the guide tissue regeneration film of compound BMSCs of the present invention is dyed for HE after rat uterus holostrome injury repair
Observation schematic；
Fig. 7 is the guide tissue regeneration film of compound BMSCs of the present invention for smooth muscle after rat uterus holostrome injury repair
Immunohistochemistry schematic diagram；
Fig. 8 is that the guide tissue regeneration film of compound BMSCs of the present invention is flat for α-SMA after rat uterus holostrome injury repair
The comparison datagram of sliding flesh content；
Fig. 9 is that the guide tissue regeneration film of compound BMSCs of the present invention is exempted from for blood vessel after rat uterus holostrome injury repair
Epidemic disease group schematic diagram；
The guide tissue regeneration film of Figure 10 compound BMSCs of the present invention is used for number of microvessels after rat uterus holostrome injury repair
Comparison datagram；
Figure 11 is the guide tissue regeneration film of compound BMSCs of the present invention for nucleus after rat uterus holostrome injury repair
It is proliferated antigen schematic diagram；
Figure 12 is that the guide tissue regeneration film of compound BMSCs of the present invention increases for cell after rat uterus holostrome injury repair
Grow several comparison datagrams；
Figure 13 is the guide tissue regeneration film of compound BMSCs of the present invention for gestation knot after rat uterus holostrome injury repair
Office's comparison schematic diagram.
Specific embodiment
Below with reference to specific embodiment and attached drawing, the present invention is described in further detail, to enable those skilled in the art
Member's refer to the instruction text can be implemented accordingly.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded one or more
A other combined presence or addition.
A kind of preparation method of the guide tissue regeneration film of compound BMSCs, comprising the following steps:
1) the BMSCs pancreatin in growth period being digested, prepares single cell suspension, the cell quantity of the single cell suspension is 2.5 ×
106A/cm2；
2) single cell suspension is taken to drop evenly on the rough surface of guide tissue regeneration film, it is spare；
3) guide tissue regeneration film that single cell suspension is added dropwise is put into temperature is 37 DEG C, and contains 5%CO2Incubator in, training
1h is supported, the guide tissue regeneration film of compound BMSCs is obtained.
The guide tissue regeneration film of the compound BMSCs of preparation is used to treat endometrial impairment, the guidance of compound BMSCs
Tissue regeneration membrane is used to prepare the composite patch for the treatment of endometrial impairment.
Endometrial impairment refers to endometrium excessive damage caused by a variety of causes, and inner membrance functionality repairs obstacle, generation
With connective tissue proliferation reparation, fibrosis, cicatrization occur for uterine wall.
The guide tissue regeneration film is provided by Shaanxi Boao Regeneration Medical Co., Ltd., guide tissue regeneration film it is main
Ingredient is collagen, and collagen is the main component of extracellular matrix, with from a wealth of sources, biocompatibility is good, it is degradable,
The advantages that preparation process is simple, guide tissue regeneration film (Fig. 1 and Fig. 2) is it has been confirmed that with the ability for promoting regeneration.
The mesenchymal stem cell is separated and is cultivated by inventor, is identified through flow cytometry cell surface antigen
(Fig. 3) and Differentiation Induction in vitro identify (Fig. 4): CD90, CD44, CD29 are positive, and CD45, CD11b are negative, can induce and are divided into rouge
Fat, bone.
A kind of guide tissue regeneration film of compound BMSCs, is prepared using above-mentioned preparation method, has irregular collagen
The rough surface surface of beam arrangement and cell covering layer, the cell covering layer are attached on rough surface surface, and between collagen beam
There is cell connection, see Fig. 5, is used to treat endometrial impairment.
A kind of sticking patch, the guide tissue regeneration film including compound BMSCs, the guide tissue regeneration film of the compound BMSCs
It is prepared using above-mentioned preparation method, the sticking patch is for treating endometrial impairment.
The concrete operations of BMSCs and guide tissue regeneration film compound criteria are as follows:
1, device therefor, material, reagent
Superclean bench, CO2Incubator, inverted microscope, centrifuge, scanning electron microscope, critical point drying instrument, Tissue Culture Flask, 6
Orifice plate, centrifuge tube, α-MEM culture solution, fetal calf serum, penicillin/streptomycin, pancreatin, glutaraldehyde, concentration gradient ethyl alcohol etc..
2, BMSCs cell activity is kept
(1) it when culture to attached cell 90% is merged, is passed on；
(2) old culture solution is absorbed, PBS is added and washes away residual culture solution, absorbs PBS；
(3) pancreatin 1ml is added, gently shakes culture bottle；
(4) it is observed under inverted microscope, discovery cytoplasm retraction, space between cells increase, and complete culture solution is added and terminates digestion；
(5) bottle wall cell is blown and beaten, cell suspension is formed it into；
(6) cell suspension is sucked into centrifuge tube, be centrifuged (room temperature 1000rpm, 5min), abandon supernatant；
(7) it is inoculated in culture bottle after adjusting cell concentration with complete culture solution, 37 DEG C, 5%CO2Incubator culture；
(8) liquid is changed every other day, continues to cultivate.
3, BMSCs and guide tissue regeneration film compound criteria
(1) guide tissue regeneration film is cut into suitably sized, smooth face-down, is placed in 6 orifice plates.
(2) well-grown BMSCs is taken, single cell suspension is prepared after pancreatin digestion, counts and adjust cell concentration.
(3) appropriate single cell suspension is taken to drop evenly on guide tissue regeneration film, cell quantity about 2.5 × 106/ cm2,
6 orifice plates are gently put into 37 DEG C, 5%CO2It is cultivated 1 hour in incubator.
4, guide tissue regeneration film provides for BMSCs and supports site
BMSCs is observed in guide tissue regeneration film surface morphology
BMSCs takes the guide tissue regeneration film for cultivating 72h as sample, with scanning with after guide tissue regeneration film compound criteria
Electronic Speculum observes cell in film surface form.
Sample is cut to area about 8 × 8mm thin slice, fixes 24 hours in 4 DEG C of 2.5% glutaraldehyde, rinses sample, gradient
Ethanol dehydration, dry in critical point drying instrument, with conducting resinl that sample dress is solid on sample stage, scanning electron microscopic observation (be shown in by result
Fig. 5).
As a result: under scanning electron microscope as it can be seen that guide tissue regeneration film rough surface surface is rendered as irregular collagen beam arrangement sample,
Surface is fused sheet of cell and covers, cell Proliferation, has cell connection between collagen beam.The result shows that BMSCs can be adhered to
In guide tissue regeneration film surface, there is good biocompatibility between BMSCs and guide tissue regeneration film.
The guide tissue regeneration film of compound BMSCs is used for serious uterine injured animal model repairing and treating:
1, BMSCs uses preceding identification
(1) appearance and microscopic observation: BMSCs and guide tissue regeneration film it is compound on the day of, to the supernatant in culture bottle into
Row color observation, under inverted microscope observe cellular morphology, cell density, whether there is or not bacterium or fungus growths.If any suspicious, palpus
Sterility test culture bacterium, mould are made in sampling.
(2) live cell fraction: the cell suspension of collection is surveyed sample on the day of BMSCs and guide tissue regeneration film are compound
Live cell fraction accounts for 95% or more.
(3) it surface marker: is measured in BMSCs and the compound first 3 days sampling row surface markers of guide tissue regeneration film: CD11b,
CD45 expression is negative；CD29, CD44, CD90 expression are positive.
2, the guide tissue regeneration film treatment rat uterus of compound BMSCs seriously damages
Choose rat uterus holostrome damage model be vulneratio uteri operate animal model --- excision be about 1.0 centimetres, it is wide by about 0.4
Centimetre holostrome uterine wall tissue.
Rat anesthesia removes abdomen center longitudinal incision and enters abdominal cavity.In uterus middle section mesometrium opposite side, excision is about
1.0cm, width about 0.4cm holostrome uterine wall tissue.Respectively by guide tissue regeneration film and the guide tissue regeneration for being compounded with BMSCs
Film repairs defect, closes abdominal incision.
Postoperative 30 days and draw materials respectively after 90 days row HE dyeing (Fig. 6) and immunohistochemical staining progress Histological assessment, art
Row pregnancy tests in 90 days carry out function assessment assessment afterwards.
As a result: postoperative 30 Tian Ji90Tian Histological assessment shows, compared with guide tissue regeneration film group, guide tissue regeneration
Film/BMSCs group endometrium and muscle layer are thicker, there is a large amount of blood vessels, body of gland and cell (see Fig. 7, Fig. 9 and Figure 11), data ratio
Fig. 8, Figure 10 and Figure 12 are seen compared with figure, it was demonstrated that guide tissue regeneration film/BMSCs is used for smooth muscle after rat uterus holostrome injury repair
Quantity, the quantity of the quantity of capilary and cell nuclear proliferating antigen be all larger than the data after guide tissue regeneration film treatment, table
Bright present invention treatment endometrial impairment has the effect of excellent.
Pregnancy tests are shown: guide tissue regeneration film group has no gravidism, and guide tissue regeneration film/BMSCs group has
Obvious gestation, and pregnancy rate obviously increases that (see Figure 13, black arrow represents embryo position, white compared with guide tissue regeneration film group
Arrow represents operative site).
As a result it prompts: endometrium and smooth after guide tissue regeneration film/BMSCs can promote rat uterus holostrome to damage
Flesh regeneration improves tissue vascularization and cellularised, promotion uterogestation functional rehabilitation.
In addition, the guide tissue regeneration film of compound BMSCs when treatment endometrial impairment, it can be with the shape of sticking patch
Formula treats endometrial impairment, determines the size of sticking patch according to the size of damage.
The present invention has found that BMSCs can be adhered to guide tissue regeneration film surface by vitro culture and zoopery, and
There is good biocompatibility with the collagen beam on guide tissue regeneration film, the compound BMSCs of guide tissue regeneration film can promote
It is repaired after into serious uterine inner film injury, increases inner film thickness, endo cell quantity and inner membrance local vascular density, it is non-toxic.
So the guide tissue regeneration film of compound BMSCs can be used for treating serious uterine inner film injury.
The present invention treats serious uterine inner film injury: active constituent of the BMSCs as treatment serious uterine inner film injury, tool
There have the advantages that obtain to be convenient, can supplement local stem cell population, and secretes growth factor improvement local microenvironment and be immunized
The effects of adjusting.Guide tissue regeneration film has good biocompatibility and safety.It can be provided for BMSCs supports position
Point improves part BMSCs concentration, extends BMSCs action time.The compound BMSCs of guide tissue regeneration film can promote intrauterine
Film smooth muscle reparation increases inner film thickness and local vascular density.
The present invention overcomes hysteroscope operation mechanicalnesses to separate Asherman's syndrom, postoperative placement of intrauterine devices or the material that prevents adhesion
Material, Yu Shuhou give estrogen and the brought endometrium of inner membrance growth are promoted seriously to damage, and can not solve inner membrance scarring problem, no
The defects of being able to achieve inner membrance functionality reparation, and easily sending out adhesion again.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With can be applied to various suitable the field of the invention entirely and be easily implemented for those skilled in the art
In addition modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is not limited to spies
Fixed details and legend shown and described herein.
Claims (10)
1. a kind of preparation method of the guide tissue regeneration film of compound BMSCs, which comprises the following steps:
The BMSCs pancreatin in growth period is digested, single cell suspension is prepared；
Single cell suspension is taken to drop evenly on guide tissue regeneration film, it is spare；
The guide tissue regeneration film that single cell suspension is added dropwise is put into incubator and is cultivated, the guide tissue regeneration of BMSCs is obtained
Film.
2. the preparation method of the guide tissue regeneration film of compound BMSCs as described in claim 1, which is characterized in that the list
The cell quantity of cell suspension is 2.5 × 106A/cm2。
3. the preparation method of the guide tissue regeneration film of compound BMSCs as described in claim 1, which is characterized in that take slender
Born of the same parents' suspension drops evenly on the rough surface of guide tissue regeneration film.
4. the preparation method of the guide tissue regeneration film of compound BMSCs as described in claim 1, which is characterized in that will be added dropwise
The guide tissue regeneration film of single cell suspension, which is put into incubator, to be cultivated, and the tool of the guide tissue regeneration film of compound BMSCs is obtained
Steps are as follows for body:
It is 37 DEG C that the guide tissue regeneration film that single cell suspension is added dropwise, which is put into temperature, and contains 5%CO2Incubator in, culture
1h obtains the guide tissue regeneration film of compound BMSCs.
5. the preparation method of the guide tissue regeneration film of the compound BMSCs as described in any one of claim 1-4, feature
It is, the guide tissue regeneration film of the compound BMSCs of preparation is for treating endometrial impairment.
6. a kind of guide tissue regeneration film of compound BMSCs, which is characterized in that using such as any one of claim 1 to 5 institute
The preparation method stated is prepared, and the guide tissue regeneration film of the compound BMSCs has the coarse of irregular collagen beam arrangement
Face surface and cell covering layer, the cell covering layer is attached on rough surface surface, and has cell connection between collagen beam.
7. the guide tissue regeneration film of compound BMSCs as claimed in claim 6, which is characterized in that the compound BMSCs's draws
Tissue regeneration membrane is led for treating endometrial impairment.
8. the guide tissue regeneration film of compound BMSCs as claimed in claim 7, which is characterized in that endometrial impairment refers to
Endometrium excessive damage, inner membrance functionality repair obstacle, instead connective tissue proliferation reparation, and fibrosis occurs for uterine wall,
One of cicatrization.
9. a kind of sticking patch, which is characterized in that the guide tissue regeneration film including compound BMSCs, the guidance group of the compound BMSCs
Regeneration membrane is knitted using the preparation method preparation as described in any one of claim 1 to 5.
10. sticking patch as claimed in claim 9, which is characterized in that the sticking patch is for treating endometrial impairment.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110898254A (en) * | 2019-12-17 | 2020-03-24 | 浙江大学 | Bioactive scaffold for repairing endometrium and improving fertility |
| CN113941032A (en) * | 2020-07-16 | 2022-01-18 | 北京中卫医正科技有限公司 | Mesenchymal stem cell composite patch and preparation method and application thereof |
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| CN110898254B (en) * | 2019-12-17 | 2021-11-30 | 浙江大学 | Bioactive scaffold for repairing endometrium and improving fertility |
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