CN108371727A - A kind of decellularized vascular matrix matrix and the application in penis thickening - Google Patents
A kind of decellularized vascular matrix matrix and the application in penis thickening Download PDFInfo
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- CN108371727A CN108371727A CN201810128697.9A CN201810128697A CN108371727A CN 108371727 A CN108371727 A CN 108371727A CN 201810128697 A CN201810128697 A CN 201810128697A CN 108371727 A CN108371727 A CN 108371727A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract
The present invention relates to a kind of decellularized vascular matrix matrix and the applications in penis thickening.In order to adapt to the operation of penis thickening, suitable built-in biological cover material is provided, the multi-steps cooperatings such as the present invention is ultrasonically treated by enzymatic treatment, surfactant, DNA degradation is handled, it is prepared for a kind of new decellularized vascular matrix host material, and it is made into built-in biological set, it is used for the treatment of premature ejaculation, small and short penis.Decellularized vascular matrix matrix of the present invention or built-in biological set do not cut off back of the body nerve for penis thickening or extension or back of the body nerve isolation modus operandi, so as to avoid the probability of psychogenic erectile dysfunction is caused to the worry of cut-out back of the body nerve due to patient.
Description
Technical field
The invention belongs to the tissue engineering technique field of bio-medical material, specifically a kind of decellularized vascular matrix matrix
And the application in penis thickening.
Background technology
There are many method of previous penis overstriking, but primarily directed to microcaulia patient, cloudy after the 1990s
Stem overstriking art is just used for the plastic aesthetic surgery of normal penile.Penis thickening is cloudy commonly used in congenital or idiopathic at present
Stem depauperation and be in microcaulia person, after wife gives birth to vaginal relaxation or to itself penis rugosity it is dissatisfied due to causing property
Quality of life is not high, is strongly required row Penis thickening to the person that improves life quality.
Injection liquid silica gel prevailing decades ago is used for overstriking penis, since liquid-state silicon gel easy tos produce rejection,
Formation tubercle, paramophia, the complication such as inguinal lymphadenopathy are disabled.80th century of the twenties mid-term, start to apply
Autologous fat injection overstriking penis, but due to the preparation of fat, the difference of injecting method and injection volume, effect is also different,
Therefore arguement is also bigger.Due to being likely to occur the absorption of fat, the formation of fatty tubercle, dissatisfied, the distal end fat of shape
Accumulation etc., it may be necessary to multiple injection, so some scholars do not approve of this method.The scholar having in recent years uses corium
The method of fat flap free grafting and full wafer dermis fat flap package, row Penis thickening, it is believed that this method advantage is
Without rejection, the destruction of adipocyte is few for autologous tissue, and corium can quickly survive, and appearance is naturally, no foreign body sensation, effect
Fruit is better than fat injection.The disadvantage is that having incisional scar for area, there is the possibility of calcification necrosis.Also scholar is using self big hidden quiet
Arteries and veins overstriking penis, advantage are no rejections, and constant anatomy is easy to operate, the disadvantage is that having incisional scar for area.Recently as
The application of synthetic material, biomaterial and tissue engineering technique, such as artificial blood vessel, expanded PTFE (PTFE), silicon
The materials such as rubber prosthese and hyaluronidase gel are used to Penis thickening.But these materials are in addition to silastic prosthesis, it is general
All over more expensive, some method operations are more complicated.Although these technologies occasionally have been reported that, specific effect not yet finds statistics
Learn number report.
De- cell built-in biological set, full name are decellularized vascular matrix matrix medical tissue built-in biological set, are derived from the mankind
The sheet of allosome or membranaceous tissue are to carry out biological and chemical processing to biological corium, removing completely is each by taking off cell technology
Cell component that kind can be identified by host, that immunological rejection can be caused, if the skin of transplanting is containing cell,
The immune response of endothelial cell may lead to vessel retraction, tissue ischemia and tissue degeneratiaon's necrosis after so transplanting, once by
Tissue, is become the matrix and holder of not cell, this material just will produce immunologic inertia, therefore not by de- cell processing
Immunological rejection can occur, while completely remaining extracellular matrix components and its three dimensions frame structure.
Based on disadvantages described above, a kind of decellularized vascular matrix matrix can be used in penis thickening of research or built-in biological set
With practical significance.
Invention content
In order to adapt to the operation of penis thickening, suitable built-in biological cover material is provided, the present invention lives by enzymatic treatment, surface
Property multi-steps cooperating, the invention such as agent supersound process, DNA degradation processing be prepared for a kind of new decellularized vascular matrix matrix
Material, and it is made into built-in biological set, the treatment for premature ejaculation, small and short penis etc..
A kind of covered the purpose of the present invention is to provide decellularized vascular matrix matrix and/or built-in biological preparation method and
Penis thickening operation method, with operation wound is small, operating time is short, safety is painless, bleeding is few, post-operative recovery is fast, anti-infective
The feature that ability is strong, compatibility is good.
To achieve the above object, the present invention provides the following technical solutions：
A kind of preparation method of decellularized vascular matrix matrix, includes the following steps：
Step 1 puts into allograft skin raw material in enzyme solutions, impregnates oscillation treatment, obtains semi-finished product A；The enzyme is phosphatide
Enzyme or the enzyme are phosphatidase and protease；
Semi-finished product A is taken out, is impregnated with physiological saline by step 2, and oscillation treatment obtains semi-finished product B；
Step 3, by semi-finished product B input surfactant solutions, ultrasonic immersion treatment obtains semi-finished product C；
Step 4 impregnates semi-finished product C with physiological saline, and oscillation treatment obtains semi-finished product D；
Step 5 fills semi-finished product D inputs in the container that DNA hydrolyzes enzyme solutions, and oscillation treatment obtains semi-finished product E；
Step 6 impregnates semi-finished product E with physiological saline, and oscillation treatment obtains semi-finished product F；
Semi-finished product F water for injection cleaning and dippings are obtained finished product G by step 7；As decellularized vascular matrix matrix；It can
With directly as penis thickening surgical material using or as preparing penis thickening surgical material
Usually, decellularized vascular matrix matrix made from the above method can be stored in physiological saline；To extend it
Storage life also may be sterilized processing (such as irradiation sterilization).
For ease of the storage and transport of above-mentioned decellularized vascular matrix matrix, extend its term of validity, further, above-mentioned side
Method is further comprising the steps of：
Step 8 takes out finished product G, packs, and sterilizing obtains finished product H；Or take out finished product G, it is placed in containing frozen-dried protective
It is freeze-dried in the solution of agent；Packaging, sterilizing, obtains finished product H.
Further, protease described in step 1 includes trypsase, bromelain, papain, dispase
It is one or more in enzyme (neutral proteinase, dispase) etc..
In some embodiments, the enzyme in abovementioned steps one is phosphatidase or phosphatidase and trypsase, bromelain
It is one or more in enzyme, papain, dispase enzymes.Further, the enzyme solutions pH value is 7.0-8.0.It is preferred that
Ground, phospholipase concentration 0.1g/L-0.4g/L and/or the protease (or in which trypsase, bromelain, pawpaw egg
White enzyme, dispase enzymes are any) a concentration of 0.1g/L-0.3g/L.Further, the phosphatidase is phospholipase A1, phosphorus
One or more, the preferred matter of phospholipase A1, phospholipase A2, phospholipase C, phospholipase D of lipase A2, phospholipase C, phospholipase D
Amount is than being 1:1:1:1.
In some embodiments, one oscillation treatment condition of abovementioned steps is oscillation 0.5-4 hours, and oscillation rotating speed is 10-
200rpm, temperature are 10-40 DEG C.
In some embodiments, the SDS (ten containing 0.1-0.3g/L in surfactant solution described in abovementioned steps three
Sodium dialkyl sulfate) or the surfactant solution in Triton X-100 (the polyethylene glycol octyls containing 0.1-0.3g/L
Phenyl ether).Further, ultrasonic soaking conditions are：40-80KHz, 100-400W, temperature is 1-20 DEG C, 3-8 minutes ultrasonic, leaching
Bubble 2-4 hours repeats 2-4 times.
In some embodiments, DNA described in abovementioned steps five hydrolyzes a concentration of 4-8g/L, pH 7.0- of enzyme solutions
8.0；Preferred concentration is 4-6g/L.
In some embodiments, oscillation treatment condition is oscillation 2-8 hours in abovementioned steps five, and oscillation rotating speed is 10-
200rpm, temperature are 10-40 DEG C.
In some embodiments, described Step 2: Step 4: step 6 oscillation treatment condition be oscillation 1-2 hours, shake
It is 80-150rpm to swing rotating speed, replaces physiological saline, continues oscillation treatment, repeats operation 4-6 times, and temperature is 1-5 DEG C.
In some embodiments, the step 7 water for injection cleaning and dipping condition is oscillation 2 hours, and oscillation rotating speed is
80-150rpm, temperature are 1-5 DEG C, replace water for injection, continue oscillation treatment, repeat operation 4-6 times.
In some embodiments, frozen-dried protective agent solution described in step 8 include phosphate buffer, hyaluronic acid and
Sugar, the wherein a concentration of 0.4-0.8mg/100mL of hyaluronic acid, sugared a concentration of 10-20mg/100mL, the sugar are seaweed
One or more of sugar, lactose, sucrose, glycerine, mannitol, sorbierite, mannose, glucose.The phosphate-buffered is molten
Liquid (PBS) can be prepared by this field conventional method, preferably 7.0 phosphate buffers of 10mmol/L pH.
The freeze-drying step is specially：It by finished product G in 5 DEG C of inlets, is kept for 1 hour, cool to -40 DEG C and keeps 1
Hour, it is warming up to -18 DEG C and is kept for 1 hour, be cooled to -35 DEG C again and kept for 1 hour, start vacuum pump, by drying box vacuum
Degree is extracted into 5-10Pa；Partition board is warming up to -30 DEG C with 2 hours, drying box vacuum degree is extracted into 1-5Pa, maintains 15h；It will with 1 hour
Partition board is warming up to 0 DEG C, and after maintaining 10h, measures pressure liter, until pressure, which rises, is less than 1pa；Partition board is warming up to 10 with 1 hour
DEG C, and 10h is maintained, pressure liter is measured, until pressure, which rises, is less than 1pa；Partition board is warming up to 25 DEG C with 0.5 hour, and maintains 8h,
Pressure liter is measured, until pressure, which rises, is less than 1pa；Preceding case vacuum degree should not be greater than 30Pa in entire drying process.
In some embodiments, abovementioned steps eight are packaged as finished product G being fitted into packaging bag, and sealing packaging finishes；
The sterilizing is that cobalt -60 irradiates, exposure dose 20-30kGy.
On the basis of common knowledge of the art, above-mentioned each optimum condition can be combined with each other each preferably to get the present invention
Example.
Allograft skin raw material sources of the present invention are in the skin of voluntary donor (such as non-living body).
The decellularized vascular matrix matrix prepared the invention also includes the above method (can also be referred to as penis thickening operation material
Material, penis built-in biological set).
The suture strength of decellularized vascular matrix matrix prepared by the present invention is 16N-18N.
The decellularized vascular matrix matrix (penis thickening surgical material, penis built-in biological set) needs advanced before use
The following operation of row：(rehydration) is completely soaked in 20 DEG C of -35 DEG C of physiological saline 15 minutes.
Can size appropriate be made in above-mentioned decellularized vascular matrix matrix according to actual needs.
The preservation condition of decellularized vascular matrix matrix of the present invention is at shady and cool drying, and relative humidity is less than or equal to 45%.
The invention also includes above-mentioned decellularized vascular matrix matrix to prepare life built in penis thickening surgical material or penis
Application in object set.
Another aspect of the present invention provides a kind of kit for the operation of penis thickening, which is characterized in that comprising above-mentioned
Decellularized vascular matrix matrix (penis thickening surgical material, penis built-in biological set).The kit can further include
Scalpel, operating scissors, haemostatic clamp, tweezers, sewing needle, 5-0 can absorb surgical thread, the ampoule bottle containing water for injection, syringe.
Another aspect of the present invention is provided is sleeved on preparation the moon using aforementioned decellularized vascular matrix matrix or aforementioned built-in biological
The purposes in material needed for the operation of stem thickening.
The needs that the present invention performs the operation according to penis thickening improve and optimize the preparation work of decellularized vascular matrix matrix
Skill.The product is the skin that human body is contributed to body, is handled by special biotechnology, host immune will can be caused to arrange in tissue
All cell clearances of reprimand reaction fall, while completely retaining extracellular matrix identical with original institutional framework；And according to need
It wants, by freeze-drying process, it is made to be more convenient for storing and transport.By constantly furtheing investigate discovery to the product, due to
While rejection ingredient is immunized in removal, former organized three-dimensional bracket structure is completely remained, as cytoskeleton,
It has the function of induced tissue generation, can be identified as autologous tissue by human tissue cell after being implanted into human body, has quickly new
Angiogenic and fibroblast grow into, and guiding cell reaches supplement, particularly quickly repairs along its collagen frame ordering growth
Purpose, it have good histocompatbility and mechanical property, can long-term existence, become a part for tissue, to
Complete the repair and reconstruction to tissue defect.
When this field carries out the operation of penis thickening using decellularized vascular matrix matrix, wrapped at away from coronary sulcus about 1cm
Skin annular incision cuts fascia superficialis, deep fascia to tunica albuginea surface successively, free up to radix penis part downwards along tunica albuginea surface, will
Prepuce of penis is press-offed.Decellularized vascular matrix matrix is prepared into built-in biological set suture on demand and is fixed on albuginea penis, far
It holds under coronary sulcus, both sides to corpora cavernosa penis and cavernous body of urethra inter-drain, layer-by-layer suture notch, penis appropriateness pressure dressing；
It needs to improve sewing density, it is therefore desirable to which decellularized vascular matrix matrix built-in biological set has higher suture strength.Meanwhile
Decellularized vascular matrix matrix or built-in biological set are planted to after human body, and the compatibility for having height with human normal tissue is needed,
This requires certain pliability.
Decellularized vascular matrix matrix of the present invention or built-in biological set disclosure satisfy that penis thickening operation demand.The present invention is logical
Cross enzymatic treatment, surfactant solution supersound process combines and carries out de- cell, can reach and take off cell effect well, and to true
The damage of scytoblastema matter is smaller, is conducive to improve tissue products mechanical performance, obtains preferable elasticity and toughness, improves tearing for tissue
It splits, suture strength, and contributes to DNA to hydrolyze and remove remaining DNA in dermal matrix in enzymatic treatment step, reduce cytotoxicity
And rejection.
To sum up, decellularized vascular matrix matrix of the invention or built-in biological set as penis thickening surgical material have with
Lower advantage：(1) for properties of product, decellularized vascular matrix matrix of the invention or built-in biological set are by process modification
Product, the mechanical performance of product is suitable for operation, suture strength 16N-18N.(2) for product effect, of the invention is de-
Whether there is or not quality after immunological rejection, rapid induction regeneration, implantation is soft for cellular allograft dermal matrix or built-in biological tackling
It is soft, without profile sense, internal compatibility is good, the holder stablized or masterplate effect.(3) decellularized vascular matrix matrix or built-in biological
Set does not cut off back of the body nerve for penis thickening or extension or back of the body nerve isolation modus operandi, so as to avoid since patient is to cut-out
It carries on the back the worry of nerve and leads to the probability of psychogenic erectile dysfunction.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..It is not specified in embodiment specific
Technology or condition person carry out according to technology or condition described in document in the art, or according to product description.It is used
Production firm person is not specified in reagent or instrument, is the conventional products that can be commercially available by regular distributor.
The preparation method of 1 decellularized vascular matrix matrix of embodiment
Allodermis Matrix raw material is put into a concentration of 0.3g/L phosphatidases and 0.1g/L trypsin solutions, enzyme solutions pH value
It is 7.0, temperature is 37 DEG C, and 100rpm vibrates 2 hours, replaces an enzyme solutions, this process of repetition is primary, obtains semi-finished product A；Phosphatide
Enzyme is phospholipase A1：Phospholipase A2：Phospholipase C：Phospholipase D is 1 according to mass ratio:1:1:1 mixture.
Semi-finished product A is taken out, puts into the container for filling normal saline solution and is impregnated with physiological saline, oscillation 3 times, every time
Oscillation 2 hours, oscillation rotating speed are 80rpm, and temperature is 2 DEG C, obtains semi-finished product B.
Semi-finished product B is put into containing in 0.2g/LTritonX-100 solution, 50KHz, 220W, ultrasound 5 minutes impregnate 3
Hour, it repeats this process 3 times, obtains semi-finished product C.
Semi-finished product C is taken out, puts into the container for filling normal saline solution and is impregnated with physiological saline, oscillation 3 times, every time
Oscillation 2 hours, temperature are 2 DEG C, and oscillation rotating speed is 80rpm, obtains semi-finished product D.
Semi-finished product D is put into and fills a concentration of 5g/L, the DNA that pH is 7.0 is hydrolyzed in enzyme solutions, and temperature is 37 DEG C, is shaken
Processing 4 hours is swung, oscillation rotating speed is 20rpm, obtains semi-finished product E.
Semi-finished product E inputs are filled in the container of normal saline solution and impregnated with physiological saline, vibrated 3 times, vibrate 2 every time
Hour, temperature is 2 DEG C, and oscillation rotating speed is 80rpm, obtains semi-finished product F.
Semi-finished product F inputs are filled in the container of water for injection and impregnated with physiological saline, vibrated 3 times, oscillation 2 is small every time
When, temperature is 2 DEG C, and oscillation rotating speed is 80rpm, obtains finished product G.
Finished product G is taken out, is fitted into packaging bag, is paved, is sealed, cobalt -60 is carried out and irradiates, exposure dose 20-30kGy,
It takes out, obtains finished product H eventually.Suture strength is 17.7N.
The preparation method of 2 decellularized vascular matrix matrix of embodiment
Allodermis Matrix raw material is put into a concentration of 0.2g/L phosphatidases and 0.2g/L trypsin solutions, enzyme solutions pH value
It is 7.0, temperature is 37 DEG C, and 100rpm vibrates 2 hours, replaces an enzyme solutions, this process of repetition is primary, obtains semi-finished product A；Phosphatide
Enzyme is phospholipase A1, phospholipase A2, phospholipase C and phospholipase D according to mass ratio 1:1:1:1 mixture.
Semi-finished product A is taken out, puts into the container for filling normal saline solution and is impregnated with physiological saline, oscillation 3 times, every time
Oscillation 2 hours, oscillation rotating speed are 80rpm, and temperature is 2 DEG C, obtains semi-finished product B.
Semi-finished product B is put into containing in 0.2g/LSDS solution, 10mMEDTA, 50KHz, 220W, ultrasound 7 minutes are impregnated
It 2 hours, repeats this process 3 times, obtains semi-finished product C.
Semi-finished product C is taken out, puts into the container for filling normal saline solution and is impregnated with physiological saline, oscillation 3 times, every time
Oscillation 1 hour, oscillation rotating speed are 100rpm, and temperature is 4 DEG C, obtains semi-finished product D.
Semi-finished product D is put into and fills a concentration of 7g/L, the DNA that pH is 7.0 is hydrolyzed in enzyme solutions, and temperature is 30 DEG C, is shaken
Processing 3 hours is swung, oscillation rotating speed is 30rpm, obtains semi-finished product E.
Semi-finished product E inputs are filled in the container of normal saline solution and impregnated with physiological saline, vibrated 3 times, vibrate 1 every time
Hour, oscillation rotating speed is 100rpm, and temperature is 4 DEG C, obtains semi-finished product F.
Semi-finished product F inputs are filled in the container of water for injection and impregnated with physiological saline, vibrated 3 times, oscillation 2 is small every time
When, temperature is 2 DEG C, and oscillation rotating speed is 80rpm, obtains finished product G.
Finished product G is taken out, is placed in frozen-dried protective agent solution；In 5 DEG C of inlets, is kept for 1 hour, cool to -40 DEG C and protect
It holds 1 hour, be warming up to -18 DEG C and kept for 1 hour, be cooled to -35 DEG C again and kept for 1 hour, start vacuum pump, drying box is true
Reciprocal of duty cycle is extracted into 5-10Pa；Partition board is warming up to -30 DEG C with 2 hours, drying box vacuum degree is extracted into 1-5Pa, maintains 15h；With 1 hour
Partition board is warming up to 0 DEG C, and after maintaining 10h, measures pressure liter, until pressure, which rises, is less than 1pa；Partition board is warming up to 1 hour
10 DEG C, and 10h is maintained, pressure liter is measured, until pressure, which rises, is less than 1pa；Partition board is warming up to 25 DEG C with 0.5 hour, and is maintained
8h measures pressure liter, until pressure, which rises, is less than 1pa；Preceding case vacuum degree should not be greater than 30Pa in entire drying process；Obtain semi-finished product
G1。
The frozen-dried protective agent solution is by the phosphate buffer of 10mmol/L pH 7.0, hyaluronic acid and trehalose group
At the wherein a concentration of 0.6mg/100mL of hyaluronic acid, a concentration of 10mg/100mL of trehalose.
Semi-finished product G1 is taken out, is fitted into packaging bag, is paved, is sealed, cobalt -60 is carried out and irradiates, exposure dose 20-
30kGy takes out, and obtains finished product H eventually.Suture strength is 16.8N.
1 decellularized vascular matrix matrix of experimental example (built-in biological set) is performed the operation for penis thickening
Operation method：Away from foreskin annular incision is made at coronary sulcus about 1cm, fascia superficialis, deep fascia to tunica albuginea table are cut successively
Face, it is free downwards until radix penis part, prepuce of penis is press-offed along tunica albuginea surface.Decellularized vascular matrix matrix is made on demand
It is standby to be fixed on albuginea penis at built-in biological set suture, distally under coronary sulcus, both sides to corpora cavernosa penis and the cavernous body of urethra
Inter-drain, layer-by-layer suture notch, penis appropriateness pressure dressing.
Post surgery treatment：
1. postoperative penis appropriateness pressure dressing 1-3 weeks.
2. Postoperative Foreskin edema situation notices that penis blood is transported.
3. inhibiting telotism, it can carry and a few days ago take orally chlorpromazine 12.5mg, 1 times/day, fenazil 25mg, 1 times/day.
4. taking out stitches within postoperative 8-10 days (absorbable thread can not also be torn open).
5. postoperative forbid sexual life 6 weeks.
Classical case 1
Xiong, male 38 years old, feel that penis is partially thin, carry out penis extending overstriking art：Away from penis head coronary sulcus about 1cm
Place, makees foreskin annular incision, cuts shallow, deep fascia successively to tunica albuginea superficial layer, dissociates downwards until radix penis part, by penis packet
Skin is press-offed.Albuginea penis is fixed in the built-in biological set suture that 2 decellularized vascular matrix matrix of embodiment is prepared on demand
Ditch is asked in layer surface, both sides to corpora cavernosa penis with the cavernous body of urethra.It is can absorb surgical thread layer-by-layer suture notch with 5-0, penis is suitable
Work as pressure dressing.
Postoperative penis Elastic bandage wrapping, daily dressing observation wound situation and skin blood fortune.
Preoperative nature week diameter 7.5cm, postoperative nature week diameter 12cm.
Postoperative wound healing in 12 days, sexual function measure normal, the postoperative tracking follow-up by being up to 46 months, telotism
Natural, strong, good appearance, penis is more apparent than operation consent thickening, and love life satisfaction is high, and male's Confidence improves, without it
Its complication.
Classical case 2
Luo, male 36 years old, feel small and short penis, carry out penis extending overstriking art：Away from penis head coronary sulcus about 1cm
Place, makees foreskin annular incision, cuts shallow, deep fascia successively to tunica albuginea superficial layer, dissociates downwards until radix penis part, by penis packet
Skin is press-offed.Albuginea penis is fixed in the built-in biological set suture that 1 decellularized vascular matrix matrix of embodiment is prepared on demand
Ditch is asked in layer surface, both sides to corpora cavernosa penis with the cavernous body of urethra.It is can absorb surgical thread layer-by-layer suture notch with 5-0, penis is suitable
Work as pressure dressing.
Postoperative penis Elastic bandage wrapping, daily dressing observation wound situation and skin blood fortune.
Preoperative nature week diameter 7.8cm, postoperative nature week diameter 12cm.
Postoperative wound healing in 14 days, sexual function measure normal.Postoperative 6-8 month follow-ups, sexual life time lengthening to 20-30
Minute, love life satisfaction is high, and sexual life harmony degree improves.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (13)
1. a kind of preparation method of decellularized vascular matrix matrix, which is characterized in that include the following steps：
Step 1 puts into allograft skin raw material in enzyme solutions, impregnates oscillation treatment, obtains semi-finished product A；The enzyme is phosphatidase, or
Enzyme described in person is phosphatidase and protease；
Semi-finished product A is taken out, is impregnated with physiological saline by step 2, and oscillation treatment obtains semi-finished product B；
Step 3, by semi-finished product B input surfactant solutions, ultrasonic immersion treatment obtains semi-finished product C；
Step 4 impregnates semi-finished product C with physiological saline, and oscillation treatment obtains semi-finished product D；
Step 5 fills semi-finished product D inputs in the container that DNA hydrolyzes enzyme solutions, and oscillation treatment obtains semi-finished product E；
Step 6 impregnates semi-finished product E with physiological saline, and oscillation treatment obtains semi-finished product F；
Semi-finished product F water for injection cleaning and dippings are obtained finished product G by step 7；Alternatively, further, it is further comprising the steps of：
Step 8 takes out finished product G, packs, and sterilizing obtains finished product H；Or take out finished product G, it is placed in containing freeze drying protectant
It is freeze-dried in solution；Packaging, sterilizing, obtains finished product H.
2. preparation method according to claim 1, which is characterized in that phosphatidase described in step 1 is phospholipase A1, phosphorus
Lipase A2, phospholipase C, phospholipase D it is one or more, it is highly preferred that the phosphatidase be phospholipase A1, phospholipase A2, phosphorus
Lipase C, phospholipase D are 1 according to mass ratio:1:1:1 mixture；And/or
The protease includes one or more in trypsase, bromelain, papain, dispase enzymes.
3. preparation method according to claim 1 or 2, which is characterized in that enzyme solutions pH value described in step 1 are 7.0-
8.0；And/or
Preferably, phospholipase concentration 0.1g/L-0.4g/L；And/or
Preferably, a concentration of 0.1g/L-0.3g/L of the protease.
4. according to claim 1-3 any one of them preparation methods, which is characterized in that step 1 oscillation treatment condition is oscillation
0.5-4 hours, oscillation rotating speed was 10-200rpm, and temperature is 10-40 DEG C.
5. according to claim 1-4 any one of them preparation methods, which is characterized in that surfactant solution described in step 3
In the Triton X-100 containing 0.1-0.3g/L in the SDS containing 0.1-0.3g/L or described surfactant solutions；
Preferably, ultrasonic soaking conditions are：40-80KHz, 100-400W, temperature is 1-20 DEG C, 3-8 minutes ultrasonic, impregnates 2-4
Hour, it repeats 2-4 times.
6. according to claim 1-5 any one of them preparation methods, which is characterized in that DNA hydrolases described in step 5 is molten
A concentration of 4-8g/L of liquid, pH 7.0-8.0；Preferred concentration is 4-6g/L；And/or
Oscillation treatment condition is oscillation 2-8 hours in the step 5, and oscillation rotating speed is 10-200rpm, and temperature is 10-40 DEG C.
7. according to claim 1-6 any one of them preparation methods, which is characterized in that described Step 2: Step 4: step
Six, oscillation treatment condition is oscillation 1-2 hours, and oscillation rotating speed is 80-150rpm, replaces physiological saline, continues oscillation treatment, weight
Multiple operation 4-6 times, temperature are 1-5 DEG C；And/or
The step 7 water for injection cleaning and dipping condition is oscillation 2 hours, and oscillation rotating speed is 80-150rpm, temperature 1-5
DEG C, water for injection is replaced, oscillation treatment is continued, repeats operation 4-6 times.
8. according to claim 1-7 any one of them preparation methods, which is characterized in that frozen-dried protective agent solution described in step 8
It is made of phosphate buffer, hyaluronic acid and sugar, wherein a concentration of 0.4-0.8mg/100mL of hyaluronic acid, sugared concentration
For 10-20mg/100mL, the sugar is in trehalose, lactose, sucrose, glycerine, mannitol, sorbierite, mannose, glucose
It is one or more of；Preferably, the phosphate buffering liquid concentration is 10mmol/L, pH 7.0.
9. according to claim 1-8 any one of them preparation methods, which is characterized in that described in step 8 freeze-drying include：
It by finished product G in 5 DEG C of inlets, is kept for 1 hour, cool to -40 DEG C and is kept for 1 hour, be warming up to -18 DEG C and kept for 1 hour, again
It is cooled to -35 DEG C to be kept for 1 hour, starts vacuum pump, drying box vacuum degree is extracted into 5-10Pa；Be warming up to partition board with 2 hours-
30 DEG C, drying box vacuum degree is extracted into 1-5Pa, maintains 15h；Partition board is warming up to 0 DEG C with 1 hour, and after maintaining 10h, measures pressure
Power liter, until pressure, which rises, is less than 1pa；Partition board is warming up to 10 DEG C with 1 hour, and maintains 10h, measures pressure liter, until pressure
It rises and is less than 1pa；Partition board is warming up to 25 DEG C with 0.5 hour, and maintains 8h, measures pressure liter, until pressure, which rises, is less than 1pa；It is whole
Preceding case vacuum degree should not be greater than 30Pa in a drying process.
10. decellularized vascular matrix matrix prepared by any one of claim 1-9 the methods；Preferably, the de- cell is different
The suture strength of body dermal matrix is 16N-18N.
11. a kind of decellularized vascular matrix matrix, which is characterized in that be that host can be caused to exempt from through completely removing by allograft skin raw material
All cells of epidemic disease rejection completely retain extracellular matrix identical with original institutional framework and are made；Preferably, institute
The suture strength for stating decellularized vascular matrix matrix is 16N-18N.
12. the decellularized vascular matrix matrix of claim 10 or 11 is being prepared built in penis thickening surgical material or penis
Application in biology set.
13. a kind of kit for the operation of penis thickening, which is characterized in that different containing the de- cell of claim 10 or 11
Body dermal matrix；Alternatively, the kit can also further include scalpel, operating scissors, haemostatic clamp, tweezers, sewing needle, 5-
One or more of 0 absorbable surgical thread, the ampoule bottle containing water for injection, syringe.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111760071A (en) * | 2020-07-01 | 2020-10-13 | 童艳梅 | Novel use of acellular dermal matrix |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104888274A (en) * | 2015-05-19 | 2015-09-09 | 暨南大学 | Acellular matrix with natural level of glycosaminoglycan and preparation and application thereof |
| CN105380685A (en) * | 2015-12-18 | 2016-03-09 | 长沙南仁医院有限公司 | Phallic allogeneic accellular patch thickening and lengthening surgery |
| CN106581770A (en) * | 2016-12-29 | 2017-04-26 | 北京桀亚莱福生物技术有限责任公司 | Dura graft, preparation method and applications in dural damage repair thereof |
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2018
- 2018-02-08 CN CN201810128697.9A patent/CN108371727B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104888274A (en) * | 2015-05-19 | 2015-09-09 | 暨南大学 | Acellular matrix with natural level of glycosaminoglycan and preparation and application thereof |
| CN105380685A (en) * | 2015-12-18 | 2016-03-09 | 长沙南仁医院有限公司 | Phallic allogeneic accellular patch thickening and lengthening surgery |
| CN106581770A (en) * | 2016-12-29 | 2017-04-26 | 北京桀亚莱福生物技术有限责任公司 | Dura graft, preparation method and applications in dural damage repair thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111760071A (en) * | 2020-07-01 | 2020-10-13 | 童艳梅 | Novel use of acellular dermal matrix |
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Denomination of invention: A decellularized allogeneic dermal matrix and its application in penile thickening Granted publication date: 20210226 Pledgee: Bank of Beijing Co.,Ltd. Hongxing Branch Pledgor: BEIJING JAYYALIFE BIOLOGICAL TECHNOLOGY Co.,Ltd. Registration number: Y2024980003489 |