CN107335095A - A kind of preparation method of medical multilayer placenta tissue implant - Google Patents
A kind of preparation method of medical multilayer placenta tissue implant Download PDFInfo
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- CN107335095A CN107335095A CN201710412764.5A CN201710412764A CN107335095A CN 107335095 A CN107335095 A CN 107335095A CN 201710412764 A CN201710412764 A CN 201710412764A CN 107335095 A CN107335095 A CN 107335095A
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- Prior art keywords
- tissue
- amnion
- layer
- face
- multilayer
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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
- A61L27/3683—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
- A61L27/3691—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 physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The invention discloses a kind of preparation method of medical multilayer placenta tissue implant, including obtain qualified placenta tissue material；Amnion and fine hair membrane tissue are isolated from placenta tissue material；Amnion and fine hair membrane tissue are handled, remove blood stains, cell and DNA, and the tissue to removing cell carries out disinfection；Lamination drying process；Cutting is completed.Medical multilayer placenta tissue implant produced by the present invention has more preferable adhesiving effect to deep wounds；Toughness is strong, is not easy to tear.
Description
Technical field
The present invention relates to medicine equipment dressing technical field, more particularly, to a kind of medical multilayer placenta tissue implant
Preparation method.
Background technology
Amnion is a kind of natural excellent biologic material, and normal amnion is thin, transparent, no blood vessel and nerve growth.It
It can clinically be used as basilar memebrane to promote epithelium to be formed, the differentiation of epithelial cell shifting property, enhancing substrate epithelial cell can be promoted to stick, prevent
Only Epithelial Cell Apoptosis, promote the Proliferation, Differentiation of epithelium；Inflammation and new vessels can be suppressed；Fibrosis can be suppressed, reach anti-scar
The effect of trace, while the myofibroblast differentiation induced by epithelial cell can also be suppressed, reduce cicatrization；It can be with
As the good support of cell migration growth, rebuild and repair for organizational project；Its no antigen, clinical practice will not arrange
Reprimand reaction, it is safe.To be preferably minimized the immunogenicity of amnion in the processing method of current domestic bioamnion, by each
Kind chemical reagent carries out de- cell processing to amnion, on the one hand causes chemical agent residue amount excessive and influences patient health；Separately
On the one hand the composition structure of amnion is also easily destroyed, causes toughness too poor and clinical demand can not be met.CN106540323A is ground
The bioamnion product of hair is largely improved in these two aspects, although but toughness strengthen, still it is aobvious it is thin, easily tear
Split.
The content of the invention
It is an object of the invention to provide a kind of preparation method of medical multilayer placenta tissue implant.
To achieve the above object, the present invention uses herein below：
A kind of preparation method of medical multilayer placenta tissue implant, comprises the following steps：
1) qualified placenta tissue material is obtained；
2) amnion and fine hair membrane tissue are isolated from placenta tissue material；
3) amnion and fine hair membrane tissue are handled, removes blood stains, cell and DNA, and the tissue to removing cell is carried out
Sterilization；
4) lamination drying process
In sequence then mould is put into baking by the amnion of sterilization and/or fine hair membrane tissue laminated multi-layer on mould
In case, 20-120 minutes are dried under 30-50 DEG C of temperature conditionss, obtain drying tissue；
The order of laminated multi-layer meets：
1. when for multilayer amnion when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion：Fibroblast cell layer is face-down, and basalis is face-up；
2. when for multilayer chorion when,
First layer chorion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：With first layer chorion；
Last layer of chorion：Fibroblast cell layer is face-down, and basalis is face-up；
3. when for multilayer amnion and chorial mixed structure when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：Fibroblast cell layer is face-down, and basalis is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion or chorion：Fibroblast cell layer is face-down, and basalis is face-up；
During such a mixed structure, must indicate dry tissue towards property；
5) corresponding size and dimension is cut into according to needs of medical treatment, completed.
Further, in step 2), the placenta tissue of acquisition is aseptically delivered into control process environment and soaked
Rinse is carried out in the sterile basin for filling 0.9-3.0wt% sodium-chloride water solutions (as rinse liquid), by amnion and chorion group
Knit and separated from other placenta tissues.
Further, in step 3), first, the sheep separated is cleaned repeatedly with 0.9-3.0wt% sodium-chloride water solutions
Film and fine hair membrane tissue are to remove blood stains；Then, the tissue for removing blood stains is immersed in 0.9-4.0wt% sodium-chloride water solutions
In, mixing 15-240 minutes are shaken at room temperature to remove cell；Afterwards, the tissue for removing cell is put into DNA solution,
Mixing 1-9 hours residual DNA content into organizing is shaken at room temperature to meet no more than 250 nanograms/every milligram of tissue (dry weight)；
Finally, the tissue for removing DNA is dipped into thimerosal the processing that carries out disinfection.The thimerosal can be ethanol, isopropanol, cross second
Acid etc..
Preferably, in step 4), in sequence by the amnion of sterilization and/or fine hair membrane tissue laminated multi-layer, first to multilayer
The upper and lower surface of membrane tissue applies 10-50psi pressure 5-30 minutes simultaneously, is placed on again on mould afterwards, processing is dried.Examine
Possible bonding effect is bad after considering lamination, now, extruding adhesion is carried out using pressure, by the fibroblast aspect of every tunic
Each tunic is consolidated as glue connection material and sticked together, processing is dried again afterwards.
Preferably, in step 4), first it is sprayed on film using glucose, sucrose solution, HESPAN, physiological saline or sterilized water
On, afterwards according still further to order laminated multi-layer on mould, processing is dried.Use glucose, sucrose solution, HESPAN, life
Reason salt solution or sterilized water are sprayed on the bonding effect that can strengthen on film between film.
The present invention has advantages below：
Medical multilayer placenta tissue implant produced by the present invention has more preferable adhesiving effect to deep wounds；Toughness
By force, it is not easy to tear.
Embodiment
In order to illustrate more clearly of the present invention, with reference to preferred embodiment, the present invention is described further.Ability
Field technique personnel should be appreciated that following specifically described content is illustrative and be not restrictive, and this should not be limited with this
The protection domain of invention.
Embodiment
A kind of preparation method of medical multilayer placenta tissue implant, comprises the following steps：
1st, placenta tissue material is obtained：Placenta tissue has drawn from i) full period, health, screens qualified caesarean delivery；Or
Ii) full period, health, conventional vaginal childbirth.The childbirth mother for contributing placenta tissue has to pass through medical treatment and Community health's history screening
And it is qualified, and the conventional infectious disease detection of the blood and tissue samples when giving a birth also has to comply with standard.The acquisition of tissue, bag
Dress and transport must all perform in strict accordance with written protocol, reach the farthest preservation original characteristic of placenta tissue.
2nd, amnion and fine hair membrane tissue are isolated from placenta tissue material：The placenta tissue of acquisition is aseptically sent
Moistened to control process environment and being immersed in the sterile basin for filling 0.9-3.0wt% sodium-chloride water solutions (as rinse liquid)
Wash, amnion and fine hair membrane tissue are separated from other placenta tissues.
3rd, amnion and fine hair membrane tissue are handled, removes blood stains, cell and DNA, and the tissue to removing cell is carried out
Sterilization：First, the amnion separated and fine hair membrane tissue are cleaned repeatedly with 0.9-3.0wt% sodium-chloride water solutions to remove blood
It is dirty；Then, the tissue for removing blood stains is immersed in 0.9-4.0wt% sodium-chloride water solutions, shakes mixing 15- at room temperature
240 minutes to remove cell；Afterwards, the tissue for removing cell is put into DNA solution (by HEPES solution, MgCl2Solution,
CaCl2Solution, purified water, DNA enzymatic mixing composition) in, mixing 1-9 hours residual DNA content into organizing is shaken at room temperature
Meet no more than 250 nanograms/every milligram of tissue (dry weight)；Finally, the tissue for removing DNA is immersed in 70% isopropanol and soaked
15-90 minutes carry out disinfection.
4th, lamination drying process：In sequence by the amnion of sterilization and/or fine hair membrane tissue laminated multi-layer on mould, so
Mould is put into mechanical convection baking oven (e.g., the UF PLUS-55 of MEMMERT companies production) afterwards, in 30-50 DEG C of temperature conditionss
Under processing is dried.In drying process, wind will directly blow to membrane tissue or be blown over indirectly from film surface.Dry completely general need
Want 20-120 minutes.During practical application, there are the framework according to product specification setting and " UP " product mark of applicant on mould
Know, multilayer film is put after mould and carries out cutting along gap between framework and cut that set product specification can be obtained.
The order of laminated multi-layer meets：
1. when for multilayer amnion when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion：Fibroblast cell layer is face-down, and basalis is face-up；
2. when for multilayer chorion when,
First layer chorion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：With first layer chorion；
Last layer of chorion：Fibroblast cell layer is face-down, and basalis is face-up；
3. when for multilayer amnion and chorial mixed structure when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：Fibroblast cell layer is face-down, and basalis is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion or chorion：Fibroblast cell layer is face-down, and basalis is face-up.
During such a mixed structure, palpus, which indicates, dries the (usable to design mark, example " UP " word as the aforementioned towards property of tissue
Sample mark dries tissue, be easy to perform the operation use when dividing tissue towards property).
Explanation：The substrate aspect of amnion is baby face, and fibroblast aspect is mother face；And chorial substrate aspect
It is mother face, and fibroblast aspect is baby face.
It is bad in view of possible bonding effect after lamination, in the step, in sequence by the amnion and/or chorion of sterilization
Laminated multi-layer is organized, simultaneously first can be applied 10-50psi pressure 5-30 minutes to the upper and lower surface of multilayer membrane tissue, put again afterwards
On mould, processing is dried.Now, carry out extruding adhesion using pressure, using the fibroblast aspect of every tunic as
Glue connection material, which consolidates each tunic, to stick together.Or first using glucose, sucrose solution, HESPAN, physiological saline or sterile
Water is sprayed on film, and afterwards according still further to order laminated multi-layer on mould, processing is dried.Using glucose, sucrose solution,
HESPAN, physiological saline or sterilized water are sprayed on the bonding effect that can strengthen on film between film.
5th, cut into corresponding size and dimension according to needs of medical treatment and packed.
Medical multilayer placenta tissue implant produced by the present invention has more active factorses, fine hair compared with individual layer bioamnion
Film contain abundant collagen component can faster wound healing, there is more preferable adhesiving effect to deep wounds；Toughness is strong, no
Easily tear, can preferably be fixed suture, facilitate clinical manipulation, solve existing individual layer bioamnion and hold in Clinical practice
Easily occur break, tear problem；Take similar de- cell handling process to obtain amnion simultaneously, ensure that multilayer embryonic tissue
The low immunogenicity of material and relatively complete constituent are implanted into, so as to have more preferable clinical efficacy.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not pair
The restriction of embodiments of the present invention, for those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, all embodiments can not be exhaustive here, it is every to belong to this hair
Row of the obvious changes or variations that bright technical scheme is extended out still in protection scope of the present invention.
Claims (5)
1. a kind of preparation method of medical multilayer placenta tissue implant, it is characterised in that comprise the following steps：
1) qualified placenta tissue material is obtained；
2) amnion and fine hair membrane tissue are isolated from placenta tissue material；
3) amnion and fine hair membrane tissue are handled, removes blood stains, cell and DNA, and the tissue to removing cell carries out disinfection；
4) lamination drying process
In sequence then mould is put into baking oven by the amnion of sterilization and/or fine hair membrane tissue laminated multi-layer on mould,
20-120 minutes are dried under 30-50 DEG C of temperature conditionss, obtain drying tissue；
The order of laminated multi-layer meets：
1. when for multilayer amnion when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion：Fibroblast cell layer is face-down, and basalis is face-up；
2. when for multilayer chorion when,
First layer chorion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：With first layer chorion；
Last layer of chorion：Fibroblast cell layer is face-down, and basalis is face-up；
3. when for multilayer amnion and chorial mixed structure when,
First layer amnion：Basalis is face-down, and fibroblast cell layer is face-up；
Intermediate layer chorion：Fibroblast cell layer is face-down, and basalis is face-up；
Intermediate layer amnion：With first layer amnion；
Last layer of amnion or chorion：Fibroblast cell layer is face-down, and basalis is face-up；
During such a mixed structure, must indicate dry tissue towards property；
5) corresponding size and dimension is cut into according to needs of medical treatment, completed.
A kind of 2. preparation method of medical multilayer placenta tissue implant according to claim 1, it is characterised in that step
2) in, the placenta tissue of acquisition is aseptically delivered to control process environment and is immersed in fill 0.9-3.0wt% chlorinations
Rinse is carried out in the sterile basin of sodium water solution, amnion and fine hair membrane tissue are separated from other placenta tissues.
A kind of 3. preparation method of medical multilayer placenta tissue implant according to claim 1, it is characterised in that step
3) in, first, the amnion separated and fine hair membrane tissue are cleaned repeatedly with 0.9-3.0wt% sodium-chloride water solutions to remove blood
It is dirty；Then, the tissue for removing blood stains is immersed in 0.9-4.0wt% sodium-chloride water solutions, shakes mixing 15- at room temperature
240 minutes to remove cell；Afterwards, the tissue for removing cell is put into DNA solution, it is small shakes mixing 1-9 at room temperature
Meet up to residual DNA content in tissue no more than 250 nanograms/every milligram of tissue；Finally, the tissue for removing DNA is dipped into and disappeared
Carry out disinfection processing in venom.
A kind of 4. preparation method of medical multilayer placenta tissue implant according to claim 1, it is characterised in that step
4) in, in sequence by the amnion of sterilization and/or fine hair membrane tissue laminated multi-layer, to the upper and lower surface of multilayer membrane tissue simultaneously first
Apply 10-50psi pressure 5-30 minutes, be placed on again on mould afterwards, processing is dried.
A kind of 5. preparation method of medical multilayer placenta tissue implant according to claim 1, it is characterised in that step
4) in, first it is sprayed on using glucose, sucrose solution, HESPAN, physiological saline or sterilized water on film, it is more according still further to order afterwards
Stacking is placed on mould, and processing is dried.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710412764.5A CN107335095A (en) | 2017-06-05 | 2017-06-05 | A kind of preparation method of medical multilayer placenta tissue implant |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710412764.5A CN107335095A (en) | 2017-06-05 | 2017-06-05 | A kind of preparation method of medical multilayer placenta tissue implant |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108514657A (en) * | 2018-04-04 | 2018-09-11 | 浙江大学 | The muscle for being sustained IGF-1 growth factors takes off the preparation method of cell material |
| CN111450320A (en) * | 2020-03-24 | 2020-07-28 | 江西省科星生物工程有限公司 | Biological tissue abdominal wall defect repairing material |
| CN112569407A (en) * | 2019-12-25 | 2021-03-30 | 北京光捷扬基健康科技有限公司 | Preparation method of placenta tissue engineering immunogen-removing skin scaffold |
| WO2024010858A1 (en) * | 2022-07-08 | 2024-01-11 | Axogen Corporation | Multi-layer amnion product, related devices, and related methods |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1757717A (en) * | 2005-07-12 | 2006-04-12 | 何伟 | Preparation method of dried active amnion |
| CN101366979A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Tissue patch and preparation method thereof |
| CN106540323A (en) * | 2016-10-27 | 2017-03-29 | 江西瑞诺健医学科技有限公司 | A kind of preparation method of bioamnion |
-
2017
- 2017-06-05 CN CN201710412764.5A patent/CN107335095A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1757717A (en) * | 2005-07-12 | 2006-04-12 | 何伟 | Preparation method of dried active amnion |
| CN101366979A (en) * | 2008-09-03 | 2009-02-18 | 陕西瑞盛生物科技有限公司 | Tissue patch and preparation method thereof |
| CN106540323A (en) * | 2016-10-27 | 2017-03-29 | 江西瑞诺健医学科技有限公司 | A kind of preparation method of bioamnion |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108514657A (en) * | 2018-04-04 | 2018-09-11 | 浙江大学 | The muscle for being sustained IGF-1 growth factors takes off the preparation method of cell material |
| CN112569407A (en) * | 2019-12-25 | 2021-03-30 | 北京光捷扬基健康科技有限公司 | Preparation method of placenta tissue engineering immunogen-removing skin scaffold |
| CN111450320A (en) * | 2020-03-24 | 2020-07-28 | 江西省科星生物工程有限公司 | Biological tissue abdominal wall defect repairing material |
| WO2024010858A1 (en) * | 2022-07-08 | 2024-01-11 | Axogen Corporation | Multi-layer amnion product, related devices, and related methods |
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