CN107233144A - Go application of the cell corneal stroma lens in treatment ophthalmology disease - Google Patents
Go application of the cell corneal stroma lens in treatment ophthalmology disease Download PDFInfo
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- CN107233144A CN107233144A CN201710403039.1A CN201710403039A CN107233144A CN 107233144 A CN107233144 A CN 107233144A CN 201710403039 A CN201710403039 A CN 201710403039A CN 107233144 A CN107233144 A CN 107233144A
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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
- A61F2/16—Intraocular lenses
- A61F2/1602—Corrective lenses for use in addition to the natural lenses of the eyes or for pseudo-phakic eyes
-
- 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
- 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
- A61L27/3641—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 site of application in the body
-
- 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
- A61L27/3687—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 the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification 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
- 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
- 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
- A61F2240/002—Designing or making customized prostheses
-
- 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
- A61L2430/16—Materials or treatment for tissue regeneration for reconstruction of eye parts, e.g. intraocular lens, cornea
Abstract
The invention provides a kind of application for removing cell corneal stroma lens in treatment ophthalmology disease, it is related to the technical field of organizational engineering.Cell corneal stroma lens are removed according to made from the method for the present invention, the cell component in corneal stroma lens can not only be effectively removed, the immunogenicity of corneal stroma lens is reduced, the biomechanical strength of corneal stroma lens is improved；More can effectively keep the original form and transparency of corneal stroma lens, it is obtained go cell corneal stroma lens have that security is higher, transparency more preferably with performance it is more stable the characteristics of.Therefore, new treatment method can be provided for clinically hyperpresbyopia, presbyopia and anisometropic correction as permanent implanted lens, for treating ophthalmology disease.
Description
Technical field
The present invention relates to organizational engineering technical field, cell corneal stroma lens are removed in treatment eye more particularly, to one kind
Application in section's disease.
Background technology
Long sight is that parallel rays enters a kind of refractive status after retina after intraocular, when the refractive power of eyeball is not enough
Or just produce long sight during its axiallength deficiency.Long sight is used as one kind of ophthalmology disease, the strong influence life matter of people
Amount, especially with the increase at age, the occurrence probability of long sight can also be raised.
Long sight is clinical common ametropia illness in eye, and the correction of hyperpresbyopia is always the problem of refractive therapy.At present
The method of distance vision correction treatment mainly includes frame eyeglasses, contact lens or refractive surgery.
Presbyopia is a kind of physiological phenomenon, and it is ametropia not to be that pathological state is also not belonging to, and is that people are stepped into after person in middle and old age must
The visual problem so occurred.With advancing age, there is the patient of presbyopia and must increase convex lens could obtaining clearly near
Eyesight.
At present, it is made more than the contact lens of implanted of synthetic material, can be according to the refractive status of different patients
The lens of the various number of degrees and model size are produced, Postoperative visual acuity recovers fast, but its drawback is also obvious, such as：It is postoperative can
Astigmatism increase can occur, the vaporific muddiness of cornea is likely to occur in early days；And its applicable disease is limited, and there is the related wind of cornea flap
Danger, is likely to occur corneal injury, the situation that visual quality declines during being chronically implanted；In addition, its security is relatively low, belong to
Permanent foreign body, is likely to occur immunological rejection during being chronically implanted.Therefore, for hyperpresbyopia, presbyopia and merging
Anisometropic patient is, it is necessary to find that indication is wider, security is higher, predictability is stronger, stability more preferably lens material.
For the material needed for corneal stroma is transplanted, good optical characteristics is not required nothing more than, is also required and human eye
Tissue has good histocompatbility, the drawbacks of synthetic material has very big in biological tissue's compatibility, and utilizes animal
The cornea of source property, then the problem of can largely alleviating the histocompatbility difference of synthetic material.
But using animal derived cornea there is also some problems, for example：(1) although the tissue compatible of animal derived cornea
Property preferably, but if its own cell removes incomplete, then can have the risk for producing immunological rejection；(2) in addition, although
Some methods for removing keratocyte are had at present, but existing cornea goes cellular matrix preparation technology to be easily caused angle
Film water suction swelling, causes arrangement of collagen fibers or conformation in matrix to change, declines corneal stroma transparency, cause original
Beginning form, curvature etc. change；(3) moreover, traditional corneal lens is typically to be made with special keratome cutting, technique
Relative coarseness, it is not smooth enough to be likely to result in the face of cutting, or lens component is lost, and predictive and security is poor, and postoperative near
Phase complication, such as astigmatism incidence is higher.
As can be seen here, the treatment method of ophthalmology disease is enriched there is provided new treatment material, for clinical practice, with weight
The meaning wanted.
In view of this, it is special to propose the present invention.
The content of the invention
It is existing to make up it is an object of the invention to provide application of the cell corneal stroma lens in treatment ophthalmology disease is gone
There is the deficiency in technology, alleviate the problem for the treatment of of eye disorders method present in prior art is short of.
It is described to remove cell cornea base the invention provides application of the cell corneal stroma lens in treatment ophthalmology disease is gone
The preparation method of matter lens includes：Corneal stroma lens with cell are subjected to cell cracking and crosslinking Treatment successively, then
Obtain described removing cell corneal stroma lens by sterilizing.
Further, the ophthalmology disease is long sight, antimetropia or presbyopia.
Further, the method for the cell cracking includes：With 0.9% physiology containing 0.1%-3%TritonX-100
Salt solution or phosphate buffer immersion 12-48h, then rinse 48-96h with 0.9% physiological saline.
Further, the method for the crosslinking includes：With 0.9% physiological saline or phosphate buffer containing crosslinking agent
Middle immersion, then rinses 24-96h with 0.9% physiological saline or phosphate buffer solution, and the crosslinking agent is 1- (3- diformazans
Aminopropyl) -3- ethyl carbodiimides, N- hydroxy thiosuccinimides, Geniposide or glutaraldehyde.
Further, the mass ratio of the usage amount of the crosslinking agent and corneal stroma lens is 1:1-1:15.
Further, before cell cracking processing, in addition to disinfect, the method for the sterilization includes：With
Containing concentration be 0.01~0.1mg/ml penicillin and concentration is 0.05~0.5mg/ml streptomysins physiological saline or phosphate
Buffer solution soaks 1-5h, is then rinsed with 0.9% physiological saline or phosphate buffer.
Further, after the crosslinking Treatment, in addition to sterilization treatment, the method for the sterilizing includes：Penetrated with γ
Line is irradiated, and irradiation dose is 20~30kGy.
Further, the corneal stroma lens with cell are obtained by full femtosecond laser technology.
Further, the preparation method includes：
Step (a), sterilization：The corneal stroma that the exact thickness with cell is obtained by full femtosecond laser technology is saturating
Mirror, with being 0.01~0.1mg/ml penicillin containing concentration and physiological saline or phosphorus that concentration is 0.05~0.5mg/ml streptomysins
Phthalate buffer soaks 1-5h, is then rinsed with 0.9% physiological saline or phosphate buffer；
Step (b), cell cracking：It is slow with 0.9% physiological saline or phosphate containing 0.1%-3%TritonX-100
Fliud flushing soaks 12-48h, then rinses 48-96h with 0.9% physiological saline or phosphate buffer；
Step (c), crosslinking：Soaked with 0.9% physiological saline or phosphate buffer containing crosslinking agent, Ran Houyong
0.9% physiological saline or phosphate buffer solution rinsing 24-96h, the crosslinking agent is 1- (3- dimethylamino-propyls) -3- second
Base carbodiimide, or N- hydroxy thiosuccinimides；The usage amount of the crosslinking agent cracks processing with the process cell
The mass ratio of corneal stroma lens is 1:1-1:15；
Step (d), sterilizing：Irradiated with gamma-rays, irradiation dose is 20~30kGy.
Remove cell corneal stroma lens according to made from the method for the present invention, can not only effectively remove in matrix cell into
Divide the immunogenicity of reduction matrix, improve the mechanical strength of cornea；It more can effectively keep the original form of corneal stroma and transparent
Degree, it is obtained go cell corneal stroma lens have that security is higher, transparency more preferably with performance it is more stable the characteristics of.Cause
This, can be that clinically hyperpresbyopia and anisometropic correction are provided as permanent implanted lens, for treating ophthalmology disease
New treatment method.
Brief description of the drawings
, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical scheme of the prior art
The accompanying drawing used required in embodiment or description of the prior art is briefly described, it should be apparent that, in describing below
Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before creative work is not paid
Put, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 removes the HE coloration result figures of cell corneal stroma lens；
Fig. 2 is cell and the HE coloration result figures of corneal stroma lens after being crosslinked；
Fig. 3 is with cell corneal stroma lens and goes the light transmittance of cell corneal stroma lens to compare figure；
Fig. 4 is the cell toxicity test result figure of cell corneal stroma lens；
Fig. 5 A are the micrograph (low power) of corneal stroma lens；
Fig. 5 B are the micrograph (high power) of corneal stroma lens；
Fig. 6 is the operation schematic diagram that corneal stroma lens are implanted into NZw corneal stroma；
Fig. 7 A are the micrograph of NZw operation cornea；
Fig. 7 B are the micrograph of NZw Post operation cornea；
Fig. 8 is the corneal thickness variation diagram of NZw perioperatively.
Embodiment
Technical scheme is clearly and completely described below in conjunction with embodiment, it is clear that described reality
It is a part of embodiment of the invention to apply example, rather than whole embodiments.Based on the embodiment in the present invention, the common skill in this area
The every other embodiment that art personnel are obtained under the premise of creative work is not made, belongs to the model that the present invention is protected
Enclose.
The invention provides go cell corneal stroma lens treatment ophthalmology disease in application, wherein, remove cell cornea
The preparation method of matrix lens includes：The corneal stroma lens of exact thickness with cell are subjected to cell cracking and friendship successively
Connection processing, then obtains cell corneal stroma lens after sterilizing.
In a preferred embodiment, ophthalmology disease is long sight.
In a preferred embodiment, ophthalmology disease is antimetropia.
In a preferred embodiment, ophthalmology disease is long sight.
In the preparation method for removing cell corneal stroma lens：
Wherein, the 0.1%-3%TritonX-100 being related in the method for cell cracking, for example can be, but be not limited to
0.1%TritonX-100,0.3%TritonX-100,0.5%TritonX-100,0.8%TritonX-100,1%
TritonX-100,1.5%TritonX-100,2%TritonX-100,2.5%TritonX-100 or 3%TritonX-
100。
Wherein, the crosslinking agent being related in the method for crosslinking for example can be, but be not limited to 1- (3- dimethylamino-propyls) -3-
Ethyl carbodiimide, either N- hydroxy thiosuccinimides or Geniposide, or glutaraldehyde.
Wherein, the usage amount for the crosslinking agent being related in the method for crosslinking is 1 with the mass ratio of corneal stroma lens:1-1:
15, its mass ratio such as can be, but be not limited to 1:1、1:2、1:3、1:4、1:5、1:6、1:7、1:8、1:9、1:10、1:10、
1:11、1:12、1:13、1:14 or 1:15.
Wherein, the concentration being related in the method for sterilization is 0.01~0.1mg/ml penicillin and concentration is 0.05~0.5mg/
The physiological saline of ml streptomysins, wherein penicillin for example can be, but be not limited to 0.01mg/ml, 0.03mg/ml, 0.05mg/
ml、0.07mg/ml、0.1mg/ml；Its streptomycin for example can be, but be not limited to 0.05mg/ml, 0.06mg/ml,
0.07mg/ml、0.08mg/ml、0.09mg/ml、0.1mg/ml、0.2mg/ml、0.3mg/ml、0.4mg/ml、0.5mg/ml。
Wherein, the dose of radiation being related in the method for sterilizing be 20~30kGy, for example can be, but be not limited to 20kGy,
21kGy, 22kGy, 23kGy, 24kGy, 25kGy, 26kGy, 27kGy, 28kGy, 29kGy or 30kGy.
Wherein, the corneal stroma lens with cell come from people or other animals (other animals for example can be, but
It is not limited to pig, ox or sheep etc.), it is preferred that the corneal stroma lens with cell come from people.
Wherein, the full femtosecond laser technology being related to, refers to utilize the small incision corneal matrix lens removal surgery of full femtosecond laser
The technology that (Small Incision Lenticule Extraction, SMILE) is cut.SMILE is to apply ultrashort pulse
Laser completes the scanning of two subpulses in cornea, makes in corneal stroma and is drawn off after eyeglass, can reduce corneal nerve
Damage and the biological influence with mechanics.
In addition, the corneal stroma lens with cell being related in the present invention refer to it is undressed, directly from people or
The fresh corneal stroma lens taken out in other animal eyes.
In order to contribute to the clearer understanding present invention, now present disclosure is carried out by specific embodiment detailed
Introduction.Pointed out as being not known, the Examination on experimental operation being related in following examples is conventional molecular biology or medical science behaviour
Make method, reagent, the instrument being related to are conventional commercial reagent or instrument.
Embodiment 1 goes the preparation method of cell corneal stroma lens
Step (a), sterilization：The corneal stroma that the exact thickness with cell is obtained by full femtosecond laser technology is saturating
Mirror, with being 0.01mg/ml penicillin containing concentration and physiological saline that concentration is 0.1mg/ml streptomysins soaks 3h, Ran Houyong
0.9% physiological saline is rinsed；
Step (b), cell cracking：24h, Ran Houyong are soaked with 0.9% physiological saline containing 0.5%TritonX-100
0.9% physiological saline rinses 96h；
Step (c), crosslinking：Soaked with 0.9% physiological saline containing crosslinking agent, then use 0.9% physiological saline
24h is rinsed, crosslinking agent is 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides (EDC)；The usage amount of crosslinking agent with through meticulous
The mass ratio of the corneal stroma lens of cellular lysate processing is 1:5；
Step (d), sterilizing：Corneal lens after crosslinking is irradiated with gamma-rays, irradiation dose is 25kGy；
It should be noted that in the present embodiment 1, the corneal stroma lens with cell come from people.
In addition, the performance for removing cell corneal stroma lens that inventor prepares to the method according to embodiment 1 is carried out
Analysis, specific experiment is as follows.
HE Coloration experiments
Respectively cell cornea base is removed to the corneal stroma lens with cell, according to what the method for embodiment 1 was prepared
Matter lens carry out HE dyeing, as a result respectively as depicted in figs. 1 and 2, take off the HE colored graphs (Fig. 1) of the corneal stroma lens after cell
In it can be seen that there are some spaces substantially without cell residue, but between matrix fiber；And handed over according to the method for embodiment 1
Going in cell corneal stroma lens HE colored graphs (Fig. 2) of being obtained after connection is then substantially not visible space, illustrates that the present invention provides
Method can effectively remove the cell in corneal stroma lens, and keep corneal stroma lens morphology stable.
Light transmittance test experience
Using spectrophotometer visible light wave range (390nm-780nm) respectively to the corneal stroma lens with cell,
What the method according to embodiment 1 was prepared goes cell corneal stroma lens to carry out light transmittance analysis, as a result as shown in figure 3, pressing
The visible light transmittance rate of what the method according to embodiment 1 was prepared go cell corneal stroma lens substantially with fresh cornea matrix
(the corneal stroma lens i.e. with cell) unanimously, illustrating the light transmission of the diagonal membrane matrix lens of the technique influences smaller, energy
It is allowed to keep preferable light transmittance.
Wherein, in Fig. 3, it is cell corneal stroma lens to remove cell corneal stroma；Fresh cornea matrix is to carry
The corneal stroma lens of cell.
Cell toxicity test
With reference to standard GB/T/T_16886.5-2003, to going the corneal lens after cell to carry out Cytotoxic evaluation, knot
Fruit is as shown in Figure 4.As seen from Figure 4, with the cell normal growth for going cell corneal stroma lens to co-culture, illustrate that this goes carefully
Born of the same parents' corneal stroma lens have good biocompatibility.
Corneal surface shape is observed
Corneal lens carry out morphology observation under the microscope, as a result as shown in Figure 5 A and 5B.It can be seen by Fig. 5 A and 5B
Go out, obtained corneal stroma rims of the lens is cut using femtosecond laser technology and has no defect, form is complete.
Zoopery
With healthy new zealand white rabbit Study of Support, cell corneal stroma lens are removed by what the method according to embodiment 1 was obtained,
NZw corneal stroma interlayer pouch is implanted into, carrying out corneal surface shape function assessment with slit-lamp microscope is detected, process is shown in
Fig. 6, as a result as shown in Fig. 7 A, Fig. 7 B and Fig. 8.This goes cell corneal stroma lens to exist it can be seen from Fig. 7 A, Fig. 7 B and Fig. 8
Internal thickness change is held essentially constant, and can be kept original state, be met the requirement of its biological mechanical property, can be angle
The clinical practice of membrane matrix lens implantation correction long sight provides basic basis.
Embodiment 2 goes the preparation method of cell corneal stroma lens
Step (a), sterilization：The corneal stroma that the exact thickness with cell is obtained by full femtosecond laser technology is saturating
Mirror, with being 0.05mg/ml penicillin containing concentration and physiological saline that concentration is 0.2mg/ml streptomysins soaks 2h, Ran Houyong
0.9% physiological saline is rinsed；
Step (b), cell cracking：48h, Ran Houyong are soaked with 0.9% physiological saline containing 0.3%TritonX-100
0.9% physiological saline rinsing 96h；
Step (c), crosslinking：Soaked, then floated with 0.9% physiological saline with 0.9% physiological saline containing crosslinking agent
Wash 24h, crosslinking agent N- hydroxy thiosuccinimides (NHS)；The usage amount of crosslinking agent and the cornea that processing is cracked by cell
The mass ratio of matrix lens is 1:9；
Step (d), sterilizing：Corneal lens after crosslinking is irradiated with gamma-rays, irradiation dose is 25kGy；
It should be noted that in the present embodiment 2, the corneal stroma lens with cell come from people.
Embodiment 3 goes the preparation method of cell corneal stroma lens
Step (a), sterilization：It is 0.1mg/ml moulds with containing concentration by the fresh corneal stroma lens with cell
Element and concentration soak 1h for the physiological saline of 0.5mg/ml streptomysins, then rinsed with 0.9% physiological saline；
Step (b), cell cracking：36h, Ran Houyong are soaked with 0.9% physiological saline containing 0.4%TritonX-100
0.9% physiological saline rinses 72h；
Step (c), crosslinking：Soaked with 0.9% physiological saline containing crosslinking agent, then use 0.9% physiological saline
24h is rinsed, crosslinking agent is 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides；The usage amount of crosslinking agent by cell with splitting
The mass ratio for solving the corneal stroma lens of processing is 1:10；
Step (d), sterilizing：Corneal lens after crosslinking is irradiated with gamma-rays, irradiation dose is 25kGy；
It should be noted that in the present embodiment 3, the corneal stroma lens with cell come from people.
It is obtained to remove cell corneal stroma lens according to the preparation method of the present invention for removing cell corneal stroma lens
With advantages below：
First, cell cracking and crosslinking are carried out in the diagonal membrane matrix lens of physiological saline environment, it is to avoid corneal stroma lens
There is water suction/dewatering state, change conformation and the arrangement of collagenous fibres, corneal stroma lens is effectively kept original
Form, thickness and curvature.The cell in diagonal membrane matrix lens is cracked and is crosslinked in this state, can not only effectively be removed
Cell component in matrix is gone, the immunogenicity of matrix is reduced, the mechanical strength of cornea is improved, more effectively corneal stroma can be kept saturating
The original form and transparency of mirror, so as to obtain the corneal stroma lens that security is higher, transparency is more stable more preferably with performance.
Secondly, value-added three-dimensional rack can be grown as keratocyte after the implantation of corneal stroma lens, cornea is thin
Born of the same parents can migrate and grow in lens, finally be combined together with auto corneal, can be clinically as permanent implanted lens
Hyperpresbyopia and anisometropic correction provide new treatment method.It can be avoided using the lens as implanted contact lens
There is the permanent foreign matter situation of synthetic material class corneal lens, it is to avoid the appearance of the phenomenon such as immunological rejection.
In addition, full femtosecond laser technology can accurately be cut so that the preparation of corneal lens is more accurate, it can avoid passing
The corneal lens that system Cutting Process is caused is lost；And the application then make use of full femtosecond laser technology to be cut, preparation technology
It is more accurate, it is predictive stronger.
According to the method being related in the present invention, what is prepared removes cell corneal stroma lens, higher, saturating with security
Lightness more preferably with performance it is more stable the characteristics of.Therefore, it is possible to be applied to a variety of ophthalmology diseases for the treatment of, it is particularly applied to control
Treat long sight and presbyopia.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, it will be understood by those within the art that：Its according to
The technical scheme described in foregoing embodiments can so be modified, or which part or all technical characteristic are entered
Row equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technology
The scope of scheme.
Claims (9)
1. go application of the cell corneal stroma lens in treatment ophthalmology disease, it is characterised in that described to remove cell corneal stroma
The preparation method of lens includes：Corneal stroma lens with cell are subjected to cell cracking and crosslinking Treatment, Ran Houjing successively
Sterilizing is crossed to obtain described removing cell corneal stroma lens.
2. application according to claim 1, it is characterised in that the ophthalmology disease is long sight, antimetropia or presbyopia.
3. the application according to claim any one of 1-2, it is characterised in that the method for the cell cracking includes：With containing
Have 0.1%-3%TritonX-100 0.9% physiological saline or phosphate buffer immersion 12-48h, then with 0.9% life
Manage saline rinse 48-96h.
4. application according to claim 3, it is characterised in that the method for the crosslinking includes：With containing crosslinking agent
Soak, then rinsed with 0.9% physiological saline or phosphate buffer solution in 0.9% physiological saline or phosphate buffer
24-96h, the crosslinking agent is 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides, N- hydroxy thiosuccinimides, capital
Buddhist nun puts down or glutaraldehyde.
5. application according to claim 4, it is characterised in that the usage amount of the crosslinking agent and the matter of corneal stroma lens
Amount is than being 1:1-1:15.
6. application according to claim 5, it is characterised in that before cell cracking processing, in addition at sterilization
Reason, the method for the sterilization includes：With being 0.01~0.1mg/ml penicillin containing concentration and concentration is 0.05~0.5mg/ml
Physiological saline or phosphate buffer the immersion 1-5h of streptomysin, are then floated with 0.9% physiological saline or phosphate buffer
Wash.
7. application according to claim 6, it is characterised in that after the crosslinking Treatment, in addition to sterilization treatment, institute
Stating the method for sterilizing includes：Irradiated with gamma-rays, irradiation dose is 20~30kGy.
8. application according to claim 7, it is characterised in that the corneal stroma lens with cell are by flying entirely
Second laser technology is obtained.
9. application according to claim 8, it is characterised in that the preparation method includes：
Step (a), sterilization：The corneal stroma lens of the exact thickness with cell will be obtained by full femtosecond laser technology, used
Containing concentration be 0.01~0.1mg/ml penicillin and concentration is 0.05~0.5mg/ml streptomysins physiological saline or phosphate
Buffer solution soaks 1-5h, is then rinsed with 0.9% physiological saline or phosphate buffer；
Step (b), cell cracking：With 0.9% physiological saline or phosphate buffer containing 0.1%-3%TritonX-100
12-48h is soaked, then 48-96h is rinsed with 0.9% physiological saline or phosphate buffer；
Step (c), crosslinking：Soaked with 0.9% physiological saline or phosphate buffer containing crosslinking agent, then with 0.9%
Physiological saline or phosphate buffer solution rinsing 24-96h, the crosslinking agent is 1- (3- dimethylamino-propyls) -3- ethyl carbon
Diimine, or N- hydroxy thiosuccinimides；The usage amount of the crosslinking agent cracks the cornea of processing with the process cell
The mass ratio of matrix lens is 1:1-1:15；
Step (d), sterilizing：Irradiated with gamma-rays, irradiation dose is 20~30kGy.
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