CN106110390A - A kind of RAPA PLGA support - Google Patents
A kind of RAPA PLGA support Download PDFInfo
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- CN106110390A CN106110390A CN201610499133.7A CN201610499133A CN106110390A CN 106110390 A CN106110390 A CN 106110390A CN 201610499133 A CN201610499133 A CN 201610499133A CN 106110390 A CN106110390 A CN 106110390A
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Abstract
The present invention relates to drug world, particularly relate to a kind of RAPA PLGA support.Utilize solvent evaporated method preparation to load the PLGA microsphere of RAPA, be further able to build PARA PLGA sustained-release micro-spheres, and in Nerve Scaffold building process, microsphere added internal stent.RAPA can be discharged with localized sustained after damage implants in early days, i.e. alleviate the secondary injury that injured nerve immunological rejection causes and alleviate again the scar tissue obstruction for neuranagenesis, avoid the side effect of systemic administration simultaneously.Reach to alleviate injured nerve secondary injury, promote peripheroneural regeneration.
Description
Technical field
The present invention relates to drug world, particularly relate to a kind of RAPA-PLGA support.
Background technology
The reparation of peripheral nerve long segment defect and replacement therapy be clinical position it is frequently necessary to faced by thorny problem.
In recent years, the design of tissue engineering method and texture's through engineering approaches peripheral nerve support is used to make great progress.Grind
Study carefully display: high bionical peripheral nerve tissue's engineering rack can be that long segment defect nerve provides falsework really, and draws
Lead neuranagenesis aixs cylinder to creep until passing through the defect stage.
Presently, there are main issue is that under normal circumstances, and peripheral nervous is because blood--the existence of nerve barrier not with
Body fluid, contacting blood, be in Immune privilege state as central nervous system.And after peripheral nervous damages, ruptures,
The neural broken ends of fractured bone and injured neuron are exposed under fluid environment, and violent inflammatory reaction locally occurs, show as with leukocyte be
Moving into of main inflammatory cell.These inflammatory cells can easily pass through the tube wall of tissue engineering bracket loose structure hence into
In internal micro-tubes structure, neural axon is caused inflammatory damage and peroxide while cleaning non-viable non-apoptotic cell and myelin fragment
Damage, have impact on the regeneration of nerve.And the hyperplasia of connective tissue can also make scar tissue fast-growth after nerve injury,
Form traumatic neuroma, hinder the regeneration of nerve further.
Rapamycin is a kind of novel macrolide immunosuppressants, blocks letter by different cytokine receptor
Number conduction, blocks the immunocyte such as T lymphocyte and by the process of G1 phase to S phase thus plays immunosuppressive effect.
Summary of the invention
The purpose of invention: in order to provide a kind of effect more preferable RAPA-PLGA support, specific purposes are shown in and are embodied as part
Multiple substantial technological effects.
In order to reach as above purpose, the present invention adopts the following technical scheme that:
Scheme one:
A kind of spinal cord and peripheral nerve injury repair materials, it is characterised in that it is RAPA-PLGA sustained-release micro-spheres, and it is to delay
Release the peripheral nervous support of rapamycin.
Scheme two:
A kind of RAPA-PLGA sustained-release micro-spheres, it is characterised in that use following preparation method, utilizes solvent evaporated method preparation dress
Carry RAPA PLGA microsphere, 2g PLGA is dissolved in 20ml dichloromethane, in medical material than 1:10(w/w) ratio put into 200mg
RAPA, fully shakes mixing；Above-mentioned oil phase is dropped in the Polyvinylalcohol solution of 0.5%, at high pressure even breast machine
Under effect, 3000rpm homogenizes 5min and obtains O/W Emulsion, and under room temperature, 400rpm continuation stirring 4h flings to organic solvent, 3000rpm
Centrifugal 5min collects the microsphere obtained, and washes 3 times, and lyophilization i.e. obtains RAPA-PLGA sustained-release micro-spheres.
Scheme three:
A kind of RAPA-PLGA support, it is characterised in that use following preparation method, uses chitosan and NTx albumen
Electronic scale respectively weighs 70mg, 280mg, is put in respectively in 20ml beaker, respectively adds 3 steaming water 10ml, adds 4% glacial acetic acid (PH3.2) 20
Microlitre, sealing is placed into 4 DEG C of constant temperature refrigerators and dissolves 24 hours；By two kinds of liquid mixing, frozen water environment uses high-speed homogenization
Machine stirs 60 minutes under the conditions of 5000r/min, according to microsphere carrying drug ratio result, is added by 26 mg RAPA-PLGA microspheres mixed
Close in liquid so that in experimental group support, the final concentration of RAPA reaches 2mg；Rapidly mixed liquor is injected after stirring diameter 3mm a little,
In the silica gel tube of a length of 20mm；With galvanized wire press from both sides stopped pipe mouth two ends, use self-made miniature velometer vertically by molding with 2 × 10-
5m/s enters in liquid nitrogen and carries out gradient stranguria of cold type；After fully entering liquid nitrogen, stand 4 hours；Silica gel tube molding is taken out, puts into pre-
In cold aluminum dish；It is transferred in vacuum drier, carries out vacuum lyophilization, after 8 hours, obtain Nerve Scaffold structure；With 1%
Genipin liquid cross-links 24 hours in 37 DEG C of calorstats, it is thus achieved that the timbering material that toughness is good, again vacuum freeze-drying.
Scheme four:
A kind of RAPA-PLGA support, it is characterised in that use following preparation method,
WeighCollagen type 150 milligrams and chitosan 75 milligrams, be dissolved in 0.05 mol/L acetum；4 degrees Celsius of perseverances
In temperature environment, stir 30 minutes with 1000 revs/min, make collagen suspension；
2g PLGA is dissolved in 20ml dichloromethane, in medical material than 1:10(w/w) ratio put into 200mg RAPA, fully shake
Swing mixing；Above-mentioned oil phase is dropped in 0.5%Polyvinylalcohol solution, under high pressure even breast machine effect, 3000rpm
Homogenizing 5min and obtain O/W Emulsion, under room temperature, 400rpm continuation stirring 4h flings to organic solvent, and 3000rpm is centrifuged 5min and collects
The microsphere arrived, washes 3 times, and lyophilization obtains RAPA-PLGA sustained-release micro-spheres；
RAPA-PLGA sustained-release micro-spheres, collagen and chitosan suspension are injected in the silica gel tube of internal diameter 2-15 millimeter, seal two
End, by 2 × 105Meter per second speed direct-axis is to immersing in condensing agent；
According to application, suspension is cut into corresponding length with silica gel tube frost thing, and lyophilizing 48 hours in lyophilization machine, lyophilizing is joined
Number is-40 degrees Celsius, 100 millitorrs；
Taking out RAPA-PLGA sustained-release micro-spheres, collagen and the chitosan stent being dried, immersing concentration is that 5-15 grams per liter genipin is molten
After cross-linking 48 hours in liquid, lyophilizing 24 hours in lyophilization machine, lyophilizing parameter is-40 degrees Celsius, 100 millitorrs.
Scheme five:
A kind of RAPA-PLGA support, it is characterised in that it is characterized in that, the micro-pipe of internal stent has the knot of axial matrix arrangement
Structure feature, micro-pipe diameter is at 20 microns～300 microns, and the micro-pipe internal surface of support is distributed RAPA-PLGA sustained-release micro-spheres.
Scheme six
Genipin strengthens the purposes of RAPA-PLGA support timbering material mechanical strength as cross-linking agent.
Scheme seven:
Genipin medicinal usage in preparing RAPA-PLGA support.
Use this utility model of as above technical scheme, have the advantages that relative to prior art: by building
PARA-PLGA sustained-release micro-spheres, and in Nerve Scaffold building process, microsphere is added internal stent.Implant in early days in damage
After can discharge RAPA with localized sustained, secondary injury that not only light injured nerve immunological rejection had caused but also alleviate scar tissue pair
In the obstruction of neuranagenesis, avoid the side effect of systemic administration simultaneously, be to use Method of Tissue Engineering to promote peripheral nervous regeneration
New trial.
Original engineered nerve repair material can substitute the material of autologous free nerve grafting, it is to avoid makes for district
Become iatrogenic infringement, accelerate the speed of peripheral nervous regeneration axonal growth, shorten the time of CO2 laser weld.But when peripheral nervous is sent out
After raw damage, fracture, the neural broken ends of fractured bone and injured neuron are exposed under fluid environment, violent inflammatory reaction locally occurs, makes
Discharge RAPA by the peripheral nervous support energy localized sustained of slow release rapamycin, both alleviated injured nerve immunological rejection and caused
Secondary injury alleviate again the scar tissue obstruction for neuranagenesis, avoid the side effect of systemic administration simultaneously.
Figure of description:
Fig. 1 is the lab diagram of the release in vitro of RAPA-PLGA microsphere support；
Fig. 2 is that postoperative nerve recovers experimental result；
Fig. 3 A-3H is fluorogold retrograde tracing result；
Fig. 4 is statistics figure and the disparity map of Fig. 3；
Fig. 5 is regenerating nerve morphological outcomes；
Fig. 6 is the significant difference figure one of Fig. 5；
Fig. 7 is the significant difference figure two of Fig. 5；
Fig. 8 is gastrocnemius PAS coloration result figure.
Detailed description of the invention
Illustrating embodiments of the invention below, embodiment is not construed as limiting the invention:
The preparation of RAPA-PLGA sustained-release micro-spheres:
Solvent evaporated method preparation is utilized to load the PLGA microsphere of RAPA.2g PLGA is dissolved in 20ml dichloromethane, by medical material ratio
Ratio 1:10(w/w) puts into 200mg RAPA, fully shakes mixing.Above-mentioned oil phase is dropped to 0.5%PVA (polyvinyl alcohol,
Polyvinylalcohol), in solution, under high pressure even breast machine effect, 3000rpm homogenizes 5min and obtains O/W Emulsion, under room temperature
400rpm continues stirring 4h and flings to organic solvent, and 3000rpm is centrifuged 5min and collects the microsphere obtained, and washes 3 times, and lyophilization is i.e.
Obtain RAPA-PLGA sustained-release micro-spheres.
Prepared by RAPA-PLGA support:
Chitosan and NTx albumen electronic scale are respectively weighed 70mg, 280mg, is put in respectively in 20ml beaker, respectively adds 3 steamings
Water 10ml, adds 4% glacial acetic acid (PH3.2) 20 microlitre, and sealing is placed into 4 DEG C of constant temperature refrigerators and dissolves 24 hours.Two kinds of liquid are mixed
Close, use high-speed homogenization machine to stir under the conditions of 5000r/min in frozen water environment 60 minutes, according to microsphere carrying drug ratio result,
26 mg RAPA-PLGA microspheres are added in mixed liquor so that in experimental group support, the final concentration of RAPA reaches 2mg.Stir a little
Mixed liquor is injected diameter 3mm, in the silica gel tube of a length of 20mm rapidly after mixing.Press from both sides stopped pipe mouth two ends with galvanized wire, use self-control micro-
Type velometer vertically by molding with 2 × 10-5M/s enters in liquid nitrogen and carries out gradient stranguria of cold type.After fully entering liquid nitrogen, stand 4
Hour.Silica gel tube molding is taken out, puts in the aluminum dish of pre-cooling.It is transferred in vacuum drier, carries out vacuum lyophilization, 8
Nerve Scaffold structure is obtained after hour.Genipin liquid with 1% cross-links 24 hours in 37 DEG C of calorstats, it is thus achieved that toughness is good
Timbering material, again vacuum freeze-drying.Subsampling is observed, through scanning electron microscopic observation, well-formed person's subpackage warp after fixing, metal spraying
Cobalt-60 illumination-based disinfection is standby.
Beneficial effect:
Original engineered nerve repair material can substitute the material of autologous free nerve grafting, it is to avoid causes doctor to for district
Source property is damaged, and accelerates the speed of peripheral nervous regeneration axonal growth, shortens the time of CO2 laser weld.But when peripheral nervous damages
After wound, fracture, the neural broken ends of fractured bone and injured neuron are exposed under fluid environment, violent inflammatory reaction locally occurs, uses slow
Releasing the peripheral nervous support energy localized sustained release RAPA of rapamycin, both alleviated that injured nerve immunological rejection causes continues
Send out damage and alleviate again the scar tissue obstruction for neuranagenesis, avoid the side effect of systemic administration simultaneously.
According to technical solution of the present invention, as a example by NTx albumen, rapamycin and chitosan, its manufacture method by with
Lower step is carried out:
1. weighCollagen type 150 milligrams and chitosan 75 milligrams, be dissolved in 0.05 mol/L acetum.At 4 degrees Celsius
In isoperibol, stir 30 minutes with 1000 revs/min, make collagen suspension；
2. 2g PLGA is dissolved in 20ml dichloromethane, in medical material than 1:10(w/w) ratio put into 200mg RAPA, fully
Concussion mixing.Above-mentioned oil phase is dropped in 0.5%PVA (polyvinyl alcohol, Polyvinylalcohol) solution, at the even breast of high pressure
Under machine effect, 3000rpm homogenizes 5min and obtains O/W Emulsion, and under room temperature, 400rpm continuation stirring 4h flings to organic solvent,
3000rpm is centrifuged 5min and collects the microsphere obtained, and washes 3 times, and lyophilization obtains RAPA-PLGA sustained-release micro-spheres.
3. RAPA-PLGA sustained-release micro-spheres, collagen and chitosan suspension are injected in the silica gel tube of internal diameter 2-15 millimeter,
Sealing both ends, by 2 × 105Meter per second speed direct-axis is to immersing in condensing agent；
4. suspension is cut into corresponding length with silica gel tube frost thing according to application, and in lyophilization machine (-40 degrees Celsius, 100
Millitorr) lyophilizing 48 hours；
5. taking out RAPA-PLGA sustained-release micro-spheres, collagen and the chitosan stent being dried, immersing concentration is 5-15 grams per liter genipin
After solution cross-links 48 hours, (-40 degrees Celsius, 100 millitorrs) lyophilizing 24 hours in lyophilization machine.
Spinal cord, peripheral nerve repairing material prepared by the present invention have the following characteristics that
1. this spinal cord, the preparation of peripheral nerve repairing material mainly use RAPA-PLGA sustained-release micro-spheres, collagen and chitosan to be main
Wanting raw material, timbering material can be that neuranagenesis provides bionical temporary structure in vivo, and can natural degradation.Without immunologic rejection and body
Interior cumulative toxicity.
2. the micro-pipe of internal stent has the architectural feature of axial matrix arrangement, and micro-pipe controlled diameter system is at 20 microns～300
Micron, beneficially Regenerating Axons oriented growth, also make the seed cell of transplanting can axially arrange along micro-tube wall, migrate, thus
The nerve making damage of limits obtains effective reparative regeneration.The outer surface of material is compact texture, can effectively stop internal
Growing into of fibrous connective tissue.Certain profile can be kept in vivo for a long time, remain to after 4 months keep preferable space to tie
Structure, this material both can be implanted directly into Bridging nerve defect, it is possible to as the carrier of seed cell plantation.
3. the micro-pipe internal surface of support is uniformly distributed a large amount of RAPA-PLGA sustained-release micro-spheres.
4. use plant extract component genipin as cross-linking agent, timbering material mechanical strength, and acellular poison can be strengthened
Property.RAPA-PLGA sustained-release micro-spheres in timbering material can play at injury region and alleviate injured nerve immunological rejection and cause
Secondary injury, alleviate the scar tissue obstruction for neuranagenesis again, avoid the side effect of systemic administration simultaneously.
Generally speaking: this patent can build PARA-PLGA sustained-release micro-spheres, and by micro-in Nerve Scaffold building process
Ball adds internal stent.RAPA can be discharged with localized sustained after damage implants in early days, i.e. alleviate injured nerve immunity row
The secondary injury scolding reaction to cause alleviates again the scar tissue obstruction for neuranagenesis, avoids the secondary work of systemic administration simultaneously
With.Reach to alleviate injured nerve secondary injury, promote peripheroneural regeneration.
It will be noted from fig. 1 that RAPA-PLGA microsphere burst effect is obvious, 24h has about 40%RAPA to discharge, base on the 5th
This discharges completely.Test 2 groups owing to RAPA being added directly into Nerve Scaffold therefore phenomenon of burst release the most substantially, after 24h
The free drug of 34% i.e. being detected, increase over time, the release profiles of RAPA stent drug tended towards stability also after 3 days
Sustained release, when the 8th day, medicine disengages completely.And test 1 group and make due to " dual-sustained-release " effect of PLGA microsphere and support
RAPA phenomenon of burst release is substantially reduced, and only has the free drug release less than 10% after 24 hours, and release profiles is mild, until the 11st
Day medicine discharges the most completely, can provide lasting slow releasing function in local in early days in nerve injury.
Fig. 1: the first group of microsphere (RAPA-PLGA) at the 6th day complete release complete, second group (RAPA+stent)
Long drug release time reaches 10 days.But use PLGA to add the 3rd group of slow-release time of support dual-sustained-release effect up to 14 days.
Respectively organizing sciatic nerve in bridge joint immediate postoperative to damage completely, sciatic nerve index the most close-100, each group is without substantially
Difference (Fig2A).Postoperative first month (4 weeks), each group nerve all has and recovers in various degree, wherein nerve autograft group
Recovering very fast with testing 1 group of function index, remaining two groups are recovered relatively slow (Fig2B).Latter two moon of art, (8 weeks) respectively organized function
Index continues to raise, and tests 1 group and nerve autograft group recovery extent the most notable (Fig2C).3 months after operation
(12weeks), autologous nerve group and experiment 1 group are already close to recovering completely, without significant difference.Remaining two experimental group is relatively poor
(Fig 2D).
Fig. 2: immediate postoperative is respectively organized sciatic nerve function index and is all almost damaged completely, postoperative 4 weeks, 8 weeks and 12 weeks
Observe sciatic nerve function index, it can be observed that test one group and recover very fast with nerve autograft group, and nothing between two groups
Significant difference, and remaining two groups of functional rehabilitation is relatively slow.
Electro physiology result
3 months after operation, nerve autograft group on motor potential wave amplitude or nerve conduction velocity result with test compared with in the of 1 group
Relatively without significant difference.Testing 2 groups and the reduction in various degree of 3 groups of motor potential wave amplitude peak values of experiment, nerve conduction velocity is earlier above
Two groups are slowed down, and the conduction cycle is longer.
Experimental group and autotransplantation group all it is observed that by the positive anterior horn motor neurons of fluorogold labelling with
And Dorsal root ganglion sensory neuron (Fig. 3 A-3H).Parallel statistical analysis, result display autotransplantation group is counted in different aspects
Labeled neurons is most, test 1 group secondly, test 3 groups minimum, all have notable between each group.
Fig. 5: the fluorogold along neural axon retrograde labeled can distinguish labelling anterior horn motor neurons (A-D)
With the dorsal root ganglion of responsible sensory function (E-H). show from labeled quantity, the cell that nerve autograft group is labeled
At most (A & E), being followed by testing one group (B & F) and two groups (C & G) of experiment, counting minimum is three groups of (D & of experiment
H).Difference the most statistically significant (Fig. 4) between group and group.
Toluidine blue staining experiment display regenerating nerve aixs cylinder quantity differs, and counts and through statistics testing result
Display autologous nerve group Regenerating Axons at most (Fig 5A), tests 1 group next (Fig 5B), and remaining two groups less.Transmission electron microscope
Result display nerve autograft group myelin is the thickest, and neural Maturity is high (Fig 5E).Test 1 group next (Fig 5F), test 2
3 groups of myelin contour structures of group and experiment are complete, and ratio is relatively fine, and Maturity is relatively low.
Fig. 5: Toluidine blue staining and transmission electron microscope results show, autotransplantation group regenerated nervous fibers quantity is more
(A), Myelin thickness compared with normal nerve fiber is without significant difference (E).Test one group of regenerated nervous fibers quantity the most more (B),
Regenerating nerve myelin Maturity and nerve autograft group are close to (F).Test no matter two groups and three groups count (C at regenerated fiber
& D) or Maturity (G & H) the most earlier above two groups there were significant differences.Significant difference is truly had between statistical result display group
(I & J)。
Autotransplantation group rat art side gastrocnemius Fiber structure is complete (Fig8A), and without atrophy, transverse section presents red aobvious
Showing that the muscle glycogen being colored enriches, muscular movement function is normal.Test 1 group (Fig 8B) and 2 groups of (Fig 8C) myoarchitecture bases
This is complete, and transverse section redness is shallower, points out the degree difference recovered due to sciatic nerve to cause muscular movement to reduce, sugar in muscle
Former deposit is less.Testing 3 groups of serious disuse atrophies of rat gastrocnemius muscles, connective tissue increases, and muscle reduces.The flesh of Lycoperdon polymorphum Vitt simultaneously
(Fig8D) is seriously gone down in the prompting motor capacity of meat transverse section.
Fig. 8: autotransplantation group rat art side gastrocnemius Fiber structure is complete (A), and without atrophy, transverse section presents red aobvious
Showing that the muscle glycogen being colored enriches, muscular movement function is normal.Test 1 group (B) and 2 groups of (C) myoarchitectures are substantially complete, cross-section
Flushing color is shallower.(D) is seriously gone down in the muscle cross-sections prompting motor capacity testing 3 groups of colors.
The ultimate principle of the present invention, principal character and advantages of the present invention have more than been shown and described.The technology of this area
Personnel, it should be recognized that the present invention is not restricted to the described embodiments, simply illustrate this described in above-described embodiment and description
Bright principle, without departing from the spirit and scope of the present invention, the present invention also has various changes and modifications, these changes
Both fall within claimed scope with improving.
Claims (7)
1. a spinal cord and peripheral nerve injury repair materials, it is characterised in that it is RAPA-PLGA sustained-release micro-spheres, it is energy
The peripheral nervous support of slow release rapamycin.
2. a RAPA-PLGA sustained-release micro-spheres, it is characterised in that use following preparation method, utilizes solvent evaporated method to prepare
Load RAPA PLGA microsphere, 2g PLGA is dissolved in 20ml dichloromethane, in medical material than 1:10(w/w) ratio put into
200mg RAPA, fully shakes mixing；Above-mentioned oil phase is dropped in the Polyvinylalcohol solution of 0.5%, at high pressure
Under the machine effect of even breast, 3000rpm homogenizes 5min and obtains O/W Emulsion, and under room temperature, 400rpm continuation stirring 4h flings to organic solvent,
3000rpm is centrifuged 5min and collects the microsphere obtained, and washes 3 times, and lyophilization i.e. obtains RAPA-PLGA sustained-release micro-spheres.
3. a RAPA-PLGA support, it is characterised in that use following preparation method, by chitosan and NTx albumen
Respectively weigh 70mg, 280mg with electronic scale, be put in respectively in 20ml beaker, respectively add 3 steaming water 10ml, add 4% glacial acetic acid (PH3.2)
20 microlitres, sealing is placed into 4 DEG C of constant temperature refrigerators and dissolves 24 hours；By two kinds of liquid mixing, use high speed even in frozen water environment
Pulp grinder stirs 60 minutes under the conditions of 5000r/min, according to microsphere carrying drug ratio result, is added by 26 mg RAPA-PLGA microspheres
In mixed liquor so that in experimental group support, the final concentration of RAPA reaches 2mg；Rapidly mixed liquor is injected diameter after stirring a little
3mm, in the silica gel tube of a length of 20mm；Press from both sides stopped pipe mouth two ends with galvanized wire, use self-made miniature velometer vertically by molding with 2 ×
10-5m/s enters in liquid nitrogen and carries out gradient stranguria of cold type；After fully entering liquid nitrogen, stand 4 hours；Silica gel tube molding is taken out, puts
Enter in the aluminum dish of pre-cooling；It is transferred in vacuum drier, carries out vacuum lyophilization, after 8 hours, obtain Nerve Scaffold structure；With
The genipin liquid of 1% cross-links 24 hours in 37 DEG C of calorstats, it is thus achieved that the timbering material that toughness is good, again vacuum freeze-drying.
4. a RAPA-PLGA support, it is characterised in that use following preparation method,
WeighCollagen type 150 milligrams and chitosan 75 milligrams, be dissolved in 0.05 mol/L acetum；At 4 degrees Celsius of constant temperature
In environment, stir 30 minutes with 1000 revs/min, make collagen suspension；
2g PLGA is dissolved in 20ml dichloromethane, in medical material than 1:10(w/w) ratio put into 200mg RAPA, fully shake
Swing mixing；Above-mentioned oil phase is dropped in 0.5%PVA (polyvinyl alcohol, Polyvinylalcohol) solution, at high pressure even breast machine
Under effect, 3000rpm homogenizes 5min and obtains O/W Emulsion, and under room temperature, 400rpm continuation stirring 4h flings to organic solvent, 3000rpm
Centrifugal 5min collects the microsphere obtained, and washes 3 times, and lyophilization obtains RAPA-PLGA sustained-release micro-spheres；
RAPA-PLGA sustained-release micro-spheres, collagen and chitosan suspension are injected in the silica gel tube of internal diameter 2-15 millimeter, seal two
End, by 2 × 105The speed direct-axis of meter per second is to immersing in condensing agent；
According to application, suspension is cut into corresponding length with silica gel tube frost thing, and lyophilizing 48 hours in lyophilization machine, lyophilizing is joined
Number is-40 degrees Celsius, 100 millitorrs；
Taking out RAPA-PLGA sustained-release micro-spheres, collagen and the chitosan stent being dried, immersing concentration is that 5-15 grams per liter genipin is molten
After cross-linking 48 hours in liquid, lyophilizing 24 hours in lyophilization machine, lyophilizing parameter is-40 degrees Celsius, 100 millitorrs.
5. a RAPA-PLGA support, it is characterised in that it is characterized in that, the micro-pipe of internal stent has axial matrix arrangement
Architectural feature, micro-pipe diameter is at 20 microns～300 microns, and the micro-pipe internal surface of support is distributed RAPA-PLGA sustained-release micro-spheres.
6. genipin strengthens the purposes of RAPA-PLGA support timbering material mechanical strength as cross-linking agent.
7. genipin medicinal usage in preparing RAPA-PLGA support.
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| US11064913B2 (en) | 2013-10-25 | 2021-07-20 | Force Impact Technologies, Inc. | Impact sensing wearable device and method |
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| US11179104B2 (en) | 2018-12-20 | 2021-11-23 | Force Impact Technologies, Inc. | Method of manufacturing mouth guard having internal components for sensing impact forces |
| US11389113B2 (en) | 2018-12-20 | 2022-07-19 | Force Impact Technologies, Inc. | Mouth guard having user-notification feature of impact force |
| US11432767B2 (en) | 2018-12-20 | 2022-09-06 | Force Impact Technologies, Inc. | Mouth guard having low-profile printed circuit board for sensing and notification of impact forces |
| US11510618B2 (en) | 2018-12-20 | 2022-11-29 | Force Impact Technologies, Inc. | Method of manufacturing mouth guard having internal components for sensing impact forces |
| US11607171B2 (en) | 2018-12-20 | 2023-03-21 | Force Impact Technologies, Inc. | Mouth guard having low-profile printed circuit board for sensing and notification of impact forces |
| US11819341B2 (en) | 2018-12-20 | 2023-11-21 | Force Impact Technologies, Inc. | Mouth guard having low-profile printed circuit board for sensing and notification of impact forces |
| US11826169B2 (en) | 2018-12-20 | 2023-11-28 | Force Impact Technologies, Inc. | Mouth guard having low-profile printed circuit board for sensing and notification of impact forces |
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Application publication date: 20161116 |