CN106264789A - Method is prepared in a kind of descemet's membrane corneal endothelial trans-plantation graft and vacuole separation thereof - Google Patents
Method is prepared in a kind of descemet's membrane corneal endothelial trans-plantation graft and vacuole separation thereof Download PDFInfo
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- CN106264789A CN106264789A CN201610885932.8A CN201610885932A CN106264789A CN 106264789 A CN106264789 A CN 106264789A CN 201610885932 A CN201610885932 A CN 201610885932A CN 106264789 A CN106264789 A CN 106264789A
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- descemet
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- vacuole
- graft
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/14—Eye parts, e.g. lenses, corneal implants; Implanting instruments specially adapted therefor; Artificial eyes
- A61F2/142—Cornea, e.g. artificial corneae, keratoprostheses or corneal implants for repair of defective corneal tissue
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2240/00—Manufacturing or designing of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2240/001—Designing or manufacturing processes
Abstract
The invention discloses a kind of descemet's membrane corneal endothelial trans-plantation graft and method is prepared in vacuole separation.The Grafted preparation method that the present invention provides, comprise the steps: with porcine cornea as raw material, by the way of injecting fluid, between hypothallus and descemet's membrane, form overall big vacuole, then cut separation, obtain the graft being made up of descemet's membrane and endothelial layer.The present invention is with Oculus sus domestica ball as raw material, it is prepared for the graft being made up of descemet's membrane and endothelial layer, the current human corneal endothelial donor present situation that there is a serious shortage in the supply can be effectively alleviated in expectation, the hope recovered lost eyesight is brought for a large amount of corneal blindness patients, there is great social value, and have the biggest industrialization, commercialization is worth, and will bring considerable economic worth after putting into clinical market.
Description
Technical field
The present invention relates to a kind of descemet's membrane corneal endothelial trans-plantation graft and method is prepared in vacuole separation, specifically relate to
And the descemet's membrane corneal endothelial trans-plantation graft prepared of a kind of porcine cornea and vacuole separation thereof prepare method.
Background technology
According to statistics, China there are about at present 700000 because of severe corneal disease the patient of blinding, corneal blindness has become as
Two treatability are blind.Corneal transplantation is the only resource that patient recovers lost eyesight.The patient of about 50% can be transplanted by corneal endothelium
Art (Endothelial Keratoplasty, EK) is recovered lost eyesight.As up-to-date corneal endothelial trans-plantation, descemet's membrane corneal endothelium
Transplantation (Descemet Membrane Endothelial Keratoplasty, DMEK) has what other EK many did not possessed
Advantage, thus potential applicability in clinical practice is wide.But, the present situation that people's corneal donor source is deficient seriously constrains DMEK development, because of
And find the bioactive materials that can substitute and become inevitable.
With tradition Penetrating Keratoplasty (Penetrating Keratoplasty, PKP) cornea holostrome transplanted to
The mode of donor is different, EK only by the endothelial layer replacement of donor's cornea to patient, this compensate for a great extent PKP some not
Evitable defect.EK mode ground constantly innovation so that it is potential applicability in clinical practice is the most wide, and up-to-date DMEK is especially with good
The recovery of postoperative visual quality, the lightest Postoperative inflammatory reaction, the relatively low unique advantage of immunologic rejection rate.The graft that DMEK requires
Only comprise descemet's membrane and corneal endothelial layer, there is the characteristic such as thickness thin (about 15 μm), fragile fragile, easy curling, thus by it
Completely separate difficulty from corneal stroma bigger.On the other hand, owing to being affected by traditional view, the making of laws and regulations
About, the public contribute the shortcoming etc. of publicity and education work, China's corneal donor source present situation that there is a serious shortage in the supply the most seriously constrains
The development of DMEK, owing to porcine cornea wide material sources, physics and optical property and endothelial cell morphology are similar to people's cornea to function,
Thus become the optimum selection of current alternative human corneal endothelial.
Cornea is divided into five layers, is followed successively by by after forward direction: epithelium layer (Epithelium), bowman's lamina
(BowmanMembrane), hypothallus (Stroma), descemet's membrane (Descemet Membrane, DM) and endothelial layer
(Endothelium).Epithelial cell thickness about 50 μm, accounts for the 10% of whole corneal thickness, is made up of 5-6 layer epithelial cell, angle
Film peripheral part epithelium layer thickens, and cell increases to 8-10 layer.Bowman's lamina is one layer of homogeneous clear film, acellular one-tenth
Point, Electron microscope showed is mainly made up of collagen fiber.Hypothallus is by about 200 layers of collagenous fiber bundle parallel with anterior corneal surface
Thin plate forms, and thickness about 500 μm accounts for the 90% of whole corneal thickness.Descemet's membrane is more tough and tensile transparent homogeneous membrane, is imbued with bullet
Property, strong to the resistance of chemical substance and pathological lesion.Endothelial layer is that one layer of hexagon pinacocyte is constituted, and has angle
Film-aqueous barrier function.
Summary of the invention
It is an object of the invention to provide a kind of descemet's membrane corneal endothelial trans-plantation graft and be prepared by vacuole separation
Method, is specifically related to descemet's membrane corneal endothelial trans-plantation graft prepared by a kind of porcine cornea and prepared by vacuole separation
Method.
The invention provides a kind of Grafted preparation method with porcine cornea as material, comprise the steps:
(1) take in vitro Oculus sus domestica ball, cut the outward flange cornea with sclerotic ring；
(2) take the cornea that step (1) obtains, remove trabecular reticulum, expose descemet's membrane not-go-end；
(3) extracting liquid with the syringe equipped with syringe needle, inject the cornea completing step (2), injection system is such as
Under:
1. needle point skin cell layer direction inserting needle in the diagonal at descemet's membrane not-go-end, needle point moves under water in hypothallus, (to the greatest extent
Amount) near descemet's membrane but descemet's membrane can not be destroyed；Fluid injection is started, when liquid enters after pinpoint inclined plane is completely into hypothallus
Accelerate to inject speed when entering to be formed between hypothallus and descemet's membrane vacuole until overall big vacuole is formed；If previous step
In, hypothallus occurs edema do not form vacuole, continues inserting needle forward and exceed area of edema, then will choose on needle point and fluid injection,
Accelerating to inject speed when liquid enters and forms vacuole between hypothallus and descemet's membrane until overall big vacuole is formed, inserting needle is deep
Degree is not to be exceeded the 1/3 of cornea transverse diameter；
If 2. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
Rapid inserting needle position the most 1. is located relatively at the point of other quadrants as inserting needle position, repeats step 1..
If 3. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
1. the rapid point of other quadrants that is 1. located relatively at step inserting needle position 2., as inserting needle position, repeats step.
If 4. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
1. the rapid point of other quadrants that is 1. 2. located relatively at step inserting needle position 3. with step, as inserting needle position, repeats step；
(4), after completing step (3), take the cornea forming overall big vacuole between hypothallus and descemet's membrane, liquid is taken out
Going out, cutting is peeled off, and obtains the graft being made up of descemet's membrane and endothelial layer.
In step (1), the sclera in the outward flange cornea with sclerotic ring is part sclera.
In step (1), outward flange with the cornea concretely outward flange of sclerotic ring with the angle of 1-2 millimeter sclerotic ring
Film.
In step (2), described " removal trabecular reticulum " can be to remove part or all of trabecular reticulum, if bullet after realizing exposing
Power layer not-go-end.It is attached with pigment on trabecular reticulum, while therefore removing trabecular reticulum, also been removed pigment.
In step (2), can first carry out Trypan Blue, then remove (concretely striking off) trabecular reticulum.
In step (2), described Trypan Blue, the whole cornea with 1-2 millimeter sclerotic ring can be carried out trypan blue
Dyeing, it is also possible to carry out Trypan Blue at the scleral sulcus of the cornea with 1-2 millimeter sclerotic ring.
After step (2) also comprising the steps: carry out Trypan Blue, before striking off trabecular reticulum, use PBS
Rinse cornea.Described PBS concretely PBS pH7.4basic (1 ×).
In step (3), described liquid is corneal storage medium, concretely Optisol GS.
In step (3), described syringe needle be angle of bend be the bending syringe needle of 90-120 °.
In step (3), in described syringe needle, pin hole is positioned on pinpoint inclined plane.
In step (3), the described syringe that concretely syringe needle and syringe are structure as a whole of the syringe equipped with syringe needle.
In step (3), the described syringe equipped with syringe needle concretely specification is the syringe of 1ml.
In step (3), the described syringe equipped with syringe needle concretely insulin syringe, such as Shu RuiTMDisposably make
Use aseptic insulin syringe.
In step (3), extract liquid with the syringe equipped with syringe needle and concretely extract with the syringe equipped with syringe needle
0.6ml-0.8ml liquid.
In step (4), the implementation that described cutting is peeled off is: being placed in by cornea on negative pressure cutting pillow, trepan cuts, stripping
From.
The present invention also protects a kind of Grafted preparation method with porcine cornea as material, comprises the steps:
(1) take in vitro Oculus sus domestica ball, cut the outward flange cornea with sclerotic ring；
(2) take the cornea that step (1) obtains, remove trabecular reticulum, expose descemet's membrane not-go-end；
(3), after completing step (2), separate descemet's membrane and the film of endothelial layer composition, obtain by descemet's membrane and interior
The pig DMEK graft of skin cell layer composition.
In step (1), the sclera in the outward flange cornea with sclerotic ring is part sclera.
In step (1), outward flange with the cornea concretely outward flange of sclerotic ring with the angle of 1-2 millimeter sclerotic ring
Film.
In step (2), described " removal trabecular reticulum " can be to remove part or all of trabecular reticulum, if bullet after realizing exposing
Power layer not-go-end.It is attached with pigment on trabecular reticulum, while therefore removing trabecular reticulum, also been removed pigment.
In step (2), can first carry out Trypan Blue, then remove (concretely striking off) trabecular reticulum.
The present invention also protects a kind of Grafted preparation method with porcine cornea as material, comprises the steps: with pig angle
Film is raw material, forms overall big vacuole, then cut separation by the way of injecting fluid between hypothallus and descemet's membrane,
Obtain the graft being made up of descemet's membrane and endothelial layer.
Described liquid is corneal storage medium, concretely Optisol GS.
The implementation that described cutting is peeled off is: being placed in by cornea on negative pressure cutting pillow, trepan cuts, and peels off.
In vitro Oculus sus domestica ball described in any of the above is the freshest the best.
The graft that method described in any of the above prepares falls within protection scope of the present invention.
The present invention also protects described graft preparing the application in the product of descemet's membrane corneal endothelial trans-plantation.
The present invention also protects a kind of product for descemet's membrane corneal endothelial trans-plantation, including described graft.
The present invention also protects described graft as the Grafted application of descemet's membrane corneal endothelial trans-plantation.
The present inventor is on the basis of lot of experiments is groped and verified, the organizational structure in conjunction with porcine cornea self is special
Point, it is provided that the graft preparation method of complete set, to for people's descemet's membrane corneal endothelial trans-plantation.
The graft using the present invention to provide carries out descemet's membrane corneal endothelial trans-plantation and has a following advantage: 1) pig angle
Heteroantigen and antigen presenting cell contained by film endothelial tissue to lack far beyond holostrome and flaggy, and this largely avoid
The super acute generation with delayed rejection；2) the porcine cornea endothelial tissue being completely in aqueous humor will be by anterior chamber one
The cytokine of row suppression immunological rejection generation and the protection of mechanism (anterior chamber's Immune Deviation state)；3) endothelium is planted sheet and is not required to
Suture is fixed, and this avoids because suture stimulates the possibility of generation new vessels induction rejection.
Pig is as the most common domestic animal, its wide material sources, cheap for clinical field.The present invention is with Oculus sus domestica
Ball is raw material, is prepared for the graft being made up of descemet's membrane and endothelial layer, and current people's cornea can be effectively alleviated in expectation
The endothelium donor present situation that there is a serious shortage in the supply, brings, for a large amount of corneal blindness patients, the hope recovered lost eyesight, and has great social value, and
Have the biggest industrialization, commercialization is worth, and will bring considerable economic worth after putting into clinical market.
Accompanying drawing explanation
Fig. 1 is the cornea after carrying out Trypan Blue at scleral sulcus.
Fig. 2, for striking off part trabecular reticulum, exposes the cornea of descemet's membrane not-go-end.
Fig. 3 is the photo of the insulin syringe that syringe needle bends to 90-120 ° of angle.
Fig. 4 be step 1. in, needle point inserting needle obliquely at Schwabe line, the photo that needle point moves under water in hypothallus.
Fig. 5 be step 1. in, hypothallus occurs edema do not form the photo of vacuole.
Fig. 6 be step 1. in, in hypothallus occur edema do not form vacuole, continue inserting needle forward and exceed area of edema, so
After will choose on needle point and fluid injection, still do not form the photo of vacuole.
Fig. 7 be step 2. in, liquid enters the photo forming vacuole between hypothallus and descemet's membrane.
Fig. 8 be step 2. in, accelerate to inject speed when liquid enters and forms vacuole between hypothallus and descemet's membrane straight
The photo formed to overall big vacuole.
Fig. 9 is the photo being placed in by cornea on negative pressure cutting pillow.
Figure 10 is the Grafted photo of pig DMEK.
Figure 11 is to examine under a microscope the photo of pig DMEK graft corneal endothelium situation after dyeing.
Detailed description of the invention
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiment
Method, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is certainly
Routine biochemistry reagent shop is commercially available.Quantitative test in following example, is respectively provided with three times and repeats experiment, and result is made even
Average.
Corneal storage medium (OptisolGS): bausch & lomb Rochester, NY；Article No. 50006-OPT.
Insulin syringe: BD Ultra-FineTMNeedle Insulin Syringe (Shu RuiTMSingle use without
Bacterium insulin syringe), 1ml, 29G × 1/2 (0.33mm × 12.7mm), syringe needle is structure as a whole with syringe, and pin hole is positioned at pin
On the pinpoint inclined plane of head.
PBS: PBS pH7.4basic (1 ×)；gibco by LifetechnologiesTM。
Overall big vacuole: i.e. cover the vacuole of the full transverse diameter of cornea.
Embodiment 1,
1, take in vitro Oculus sus domestica ball, cut the outward flange cornea with 1-2 millimeter sclerotic ring.
2, take the cornea that step 1 obtains, at scleral sulcus, carry out Trypan Blue (in practical operation, it is also possible to step 1
The whole cornea obtained carries out Trypan Blue), then rinse with PBS, then strike off trabecular reticulum with anodontia microforceps
(being attached with pigment on trabecular reticulum), exposes descemet's membrane not-go-end (Schwabe line).
Cornea after carrying out Trypan Blue at scleral sulcus is shown in Fig. 1.
Striking off trabecular reticulum, the cornea exposing descemet's membrane not-go-end is shown in Fig. 2.
3, take insulin syringe, extract 0.6ml corneal storage medium, then syringe needle is bent to 90-120 ° of angle.
Fig. 3 is shown in by the photo of the insulin syringe that syringe needle bends to 90-120 ° of angle.
4, after completing step 3, take described insulin syringe, the cornea completing step 2 is injected that (cornea is positioned over
In culture dish, endothelial layer is upwards), injection system is as follows:
1. at Schwabe line needle point inserting needle obliquely (needle point refers to needle point skin cell layer side in the diagonal obliquely
To), needle point moves under water in hypothallus, close proximity to descemet's membrane but descemet's membrane can not be destroyed；When pinpoint inclined plane completely into
Starting fluid injection after hypothallus, liquid, in local accumulation, is accelerated when liquid enters and forms vacuole between hypothallus and descemet's membrane
Inject speed until overall big vacuole is formed；If in previous step, hypothallus occurs edema do not form vacuole, continue forward
Inserting needle exceedes area of edema, then will choose on needle point and fluid injection, forms vacuole when liquid enters between hypothallus and descemet's membrane
Time accelerate to inject speed until overall big vacuole is formed, depth of needle is not to be exceeded the 1/3 of cornea transverse diameter.
If 2. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
Rapid inserting needle position the most 1. is located relatively at the point of other quadrants as inserting needle position, repeats step 1..
If 3. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
1. the rapid point of other quadrants that is 1. located relatively at step inserting needle position 2., as inserting needle position, repeats step.
If 4. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select and walk
1. the rapid point of other quadrants that is 1. 2. located relatively at step inserting needle position 3. with step, as inserting needle position, repeats step.
Step 1. in, needle point inserting needle obliquely at Schwabe line, Fig. 4 is shown in by the photo that needle point moves under water in hypothallus.
Step 1. in, hypothallus occurs edema do not form the photo of vacuole and sees Fig. 5.
Step 1. in, in hypothallus occur edema do not form vacuole, continue inserting needle forward and exceed area of edema, then by pin
Choosing on point and fluid injection, Fig. 6 is shown in by the photo still not forming vacuole.
Step 2. in, liquid enters and forms the photo of vacuole between hypothallus and descemet's membrane and see Fig. 7.
Step 2. in, accelerate to inject speed when liquid enters and forms vacuole between hypothallus and descemet's membrane until overall
Fig. 8 is shown in by the photo that big vacuole is formed.
5, after completing step 4, take the cornea forming overall big vacuole between hypothallus and descemet's membrane, will be formed overall big
The liquid of vacuole is extracted out, is subsequently placed on negative pressure cutting pillow, carries out trepan cutting, peel off with anodontia microforceps, obtain by rear bullet
Power layer and the pig DMEK graft of endothelial layer composition.
Fig. 9 is shown in by the photo being placed in by cornea on negative pressure cutting pillow.
Figure 10 is shown in by the Grafted photo of pig DMEK.
Carry out three times repeating test, repeat test carries out aforesaid operations, pig DMEK to 100 in vitro Oculus sus domestica balls every time
Grafted yield is 85%-90%.
Comparative example 1,
1, take in vitro Oculus sus domestica ball, cut the outward flange cornea with 1-2 millimeter sclerotic ring.
2, take insulin syringe, extract 0.6ml corneal storage medium, then syringe needle is bent to 90-120 ° of angle.
3, after completing step 2, taking described insulin syringe, the cornea obtaining step 1 injects that (cornea is positioned over
In culture dish, endothelial layer is upwards), injection system is as follows:
Needle point inserting needle obliquely (needle point refers to needle point skin cell layer direction in the diagonal obliquely) at Schwabe line,
Needle point moves under water in hypothallus, close proximity to descemet's membrane but descemet's membrane can not be destroyed；When pinpoint inclined plane is completely into substrate
Starting fluid injection after Ceng, liquid, in local accumulation, is accelerated to inject when liquid enters and forms vacuole between hypothallus and descemet's membrane
Speed is until overall big vacuole is formed.
4, after completing step 3, take the cornea forming overall big vacuole between hypothallus and descemet's membrane, will be formed overall big
The liquid of vacuole is extracted out, is subsequently placed on negative pressure cutting pillow, carries out trepan cutting, peel off with anodontia microforceps, obtain by rear bullet
Power layer and the pig DMEK graft of endothelial layer composition.
Carry out three times repeating test, repeat test carries out aforesaid operations, pig DMEK to 100 in vitro Oculus sus domestica balls every time
Grafted yield is 0%-5%.
Embodiment 2,
The pig DMEK graft of Example 1 preparation, detection cell density and live cell fraction.
Concrete grammar is as follows:
1, take pig DMEK graft, use 0.2% Trypan Blue liquid dyeing 2min.
2, after completing step 1, clean 2 times with PBS.
3, after completing step 2, with 1% alizarin red S dyeing liquor dyeing 3min.
4, after completing step 3, clean 2 times with PBS.
5, after completing step 4, examining under a microscope, counting cell density (takes 3 high power field countings, summation at random
Average) and live cell fraction.
Figure 11 is shown in by photo.Arrow mark dead cell.
The pig DMEK graft of Example 1 preparation, all reaches following standard: cell density > 2500/mm2, living cells ratio
Example > 90%.
The graft of Example 1 preparation, carries out descemet's membrane corneal endothelial trans-plantation to Rhesus Macacus, Continuous Observation half
Year, Rhesus Macacus animation is good, edema cornea recovers transparent, and DMEK graft position is normal, has no that obvious rejection is sent out
Raw.
Claims (10)
1. a Grafted preparation method, comprises the steps:
(1) take in vitro Oculus sus domestica ball, cut the outward flange cornea with sclerotic ring；
(2) take the cornea that step (1) obtains, remove trabecular reticulum, expose descemet's membrane not-go-end；
(3) extracting liquid with the syringe equipped with syringe needle, inject the cornea completing step (2), injection system is as follows:
1. needle point skin cell layer direction inserting needle in the diagonal at descemet's membrane not-go-end, needle point moves under water in hypothallus, near rear bullet
Power layer but descemet's membrane can not be destroyed；After pinpoint inclined plane is completely into hypothallus start fluid injection, when liquid enter hypothallus and
Accelerate to inject speed when forming vacuole between descemet's membrane until overall big vacuole is formed；If in previous step, in hypothallus
Occur edema not form vacuole, continue inserting needle forward and exceed area of edema, then will choose on needle point and fluid injection, when liquid enters base
Accelerating to inject speed when forming vacuole between matter layer and descemet's membrane until overall big vacuole is formed, depth of needle is not to be exceeded angle
The 1/3 of film transverse diameter；
If 2. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select with step 1.
Inserting needle position be located relatively at the point of other quadrants as inserting needle position, repeat step 1..
If 3. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select with step 1.
With the point that step inserting needle position 2. is located relatively at other quadrants as inserting needle position, repeat step 1..
If 4. step does not the most obtain being formed between hypothallus and descemet's membrane the cornea of overall big vacuole, select with step 1.
2. it is located relatively at the point of other quadrants with step inserting needle position 3. with step as inserting needle position, repeats step 1.；
(4), after completing step (3), take the cornea forming overall big vacuole between hypothallus and descemet's membrane, liquid is extracted out, cuts
Cut stripping, obtain the graft being made up of descemet's membrane and endothelial layer.
2. the method for claim 1, it is characterised in that: described liquid is corneal storage medium.
3. method as claimed in claim 1 or 2, it is characterised in that: described syringe needle be angle of bend be the needle bent of 90-120 °
Head.
4. the method as described in claim 1 or 2 or 3, it is characterised in that: the implementation that described cutting is peeled off is: by cornea
Being placed on negative pressure cutting pillow, trepan cuts, and peels off.
5. a Grafted preparation method, comprises the steps:
(1) take in vitro Oculus sus domestica ball, cut the outward flange cornea with sclerotic ring；
(2) take the cornea that step (1) obtains, remove trabecular reticulum, expose descemet's membrane not-go-end；
(3), after completing step (2), separate descemet's membrane and the film of endothelial layer composition, obtain by descemet's membrane and endothelium thin
The pig DMEK graft of born of the same parents' layer composition.
6. a Grafted preparation method, comprises the steps: with porcine cornea as raw material, at base by the way of injecting fluid
Form overall big vacuole between matter layer and descemet's membrane, then cut separation, obtain being made up of descemet's membrane and endothelial layer
Graft.
7. the graft that in claim 1 to 6, arbitrary described method prepares.
8. the application in preparation is used for the product of descemet's membrane corneal endothelial trans-plantation of the graft described in claim 7.
9. for a product for descemet's membrane corneal endothelial trans-plantation, including graft described in claim 7.
10. graft described in claim 7 is as the Grafted application of descemet's membrane corneal endothelial trans-plantation.
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Cited By (3)
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| CN106955372A (en) * | 2017-04-12 | 2017-07-18 | 山东省眼科研究所 | A kind of construction method of tissue engineering comea endothelium |
| RU2647828C1 (en) * | 2017-04-10 | 2018-03-19 | Федеральное государственное бюджетное научное учреждение "Научно-исследовательский институт глазных болезней" | Method of posterior layer-penetrating keratoplasty in treating bullous keratopathy |
| CN112603646A (en) * | 2020-12-21 | 2021-04-06 | 上海昶衡生物科技有限公司 | Suprachoroidal space syringe |
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| US20110166650A1 (en) * | 2008-09-04 | 2011-07-07 | Massimo Busin | New technique for preparing, storing and transplanting endothelial grafts |
| US20140012295A1 (en) * | 2012-07-06 | 2014-01-09 | Centre Hospitalier Universitaire De Rouen | Blade for preparing an endothelial graft, and associated preparation method |
| CN105636560A (en) * | 2013-10-28 | 2016-06-01 | 视乐有限公司 | Technique for laser-cutting an endothelial corneal graft |
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| RU2647828C1 (en) * | 2017-04-10 | 2018-03-19 | Федеральное государственное бюджетное научное учреждение "Научно-исследовательский институт глазных болезней" | Method of posterior layer-penetrating keratoplasty in treating bullous keratopathy |
| CN106955372A (en) * | 2017-04-12 | 2017-07-18 | 山东省眼科研究所 | A kind of construction method of tissue engineering comea endothelium |
| CN106955372B (en) * | 2017-04-12 | 2020-08-14 | 山东省眼科研究所 | Method for constructing tissue engineering corneal endothelium |
| CN112603646A (en) * | 2020-12-21 | 2021-04-06 | 上海昶衡生物科技有限公司 | Suprachoroidal space syringe |
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