CN1768856A - Method for preparing injection for removing wrinkles and scars - Google Patents
Method for preparing injection for removing wrinkles and scars Download PDFInfo
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- CN1768856A CN1768856A CNA2004100888029A CN200410088802A CN1768856A CN 1768856 A CN1768856 A CN 1768856A CN A2004100888029 A CNA2004100888029 A CN A2004100888029A CN 200410088802 A CN200410088802 A CN 200410088802A CN 1768856 A CN1768856 A CN 1768856A
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- injection
- wrinkle
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Abstract
The invention provides the process for preparing injection for scar-dispeling, which comprises, cutting skin tissues from patients, placing the skin tissures into incubator for expansion culture, charged the cultured desmocyte into standard injection by the ratio of 9.0-9.8 million/ml, thus obtaining cell suspension.
Description
Technical field
The present invention relates to tissue engineering technique, relate in particular to a kind of preparation method that is used for the injection of Wrinkle-and scar-removing.
Background technology
In the prior art, the injection of employed Wrinkle-and scar-removing is 1,000 ten thousand-4,000 ten thousand to add 5% Glucose Liquid and form by concentration.In the preparation method of this injection, the cultivation multiselect suspension method of primary cell, the shortcoming of this method is that success rate is not high.
Summary of the invention
The objective of the invention is provides a kind of injection at above-mentioned problems of the prior art, has good Wrinkle-and scar-removing effect through clinical trial.
According to above-mentioned purpose of the present invention, the preparation method that is used for the injection of Wrinkle-and scar-removing of the present invention comprises:
A. take off skin histology on one's body from the patient;
B. the skin histology that is taken off is put into incubator and carry out amplification cultivation;
C. the fibroblast of the amplification cultivation ratio in 9,000,000 every milliliter-9,800,000 is added in the standard liquid injection, make cell suspension.
Described steps A further comprises: the iodine piece disinfectant solution with 5% carries out local surfaces with 2% lignocaine then and anaesthetizes behind ear or the inboard sterilization of arm.Take off with the holostrome skin of cutisector with the 3-4 square millimeter, put into tissue and preserve liquid, wound site is sewed up, bandage covers.
Described step B further comprises:
B1. incubator digestion 10 minutes is put in the piece of tissue and the tissue fluid that obtain;
B2. remove epidermis and subepidermal fat;
B3. piece of tissue is cut into some, and with some block organizations of being cut into being mutually that six well culture plates are put at 0.5 centimetre interval;
B4. six well culture plates are carefully put into incubator, add 0.5 milliliter of culture fluid gently along the tank wall of incubator after 4 hours, add 1 milliliter of culture fluid after 24 days again;
B5. piece of tissue is gone down to posterity and cultivate 4-6 week.
Described step B5 further comprises: changed liquid once every 2-3 days, go down to posterity after 50%-70% compiles cell when cell reaches.
Preparation method of the present invention adopts tissue culture method, has obtained higher success rate.
Description of drawings
Fig. 1 shows the flow chart of preparation injection of the present invention.
The specific embodiment
Understand and realization the present invention for the ease of persons skilled in the art, describe the present invention below in detail.
One, the extraction of autogenous cell
Iodine piece disinfectant solution with 5% carries out local surfaces with 2% lignocaine then and anaesthetizes behind ear or the inboard sterilization of arm.Take off with the holostrome skin of cutisector with the 3-4 square millimeter, put into tissue and preserve in the liquid (pancreatin of DMEM+10% hyclone+0.25%, U.S. Hyclone company), wound site is sewed up, bandage covers.
Two, the preparation of injection
As shown in Figure 1, in step 1, incubator (SanyaCO2 incubator) digestion 10 minutes is put in the piece of tissue and the tissue fluid that obtain.In step 2, remove epidermis and subepidermal fat then.In step 3, piece of tissue is cut into some (every is about the 1-2 square millimeter), and with some block organizations of being cut into being mutually that six well culture plates are put at 0.5 centimetre interval.In step 4, six well culture plates are carefully put into incubator.In step 5, added 0.5 milliliter of culture fluid gently along the tank wall of incubator afterwards, and in step 6, added 1 milliliter of culture fluid after 24 days again in 4 hours.
Cultivate and to see having cell to extend out in 24 hours, had a large amount of cells to come out and begin to breed in 48 hours from marginal growth from the piece of tissue edge.Cultivate after 72 hours and in 37 ℃ of incubators, digested 10-15 minute, the routine cultivation of going down to posterity then with 0.25% pancreatin.
In step 7, go down to posterity to cultivate and changed liquid once every 2-3 days, when reaching, cell goes down to posterity after 50%-70% compiles cell.In step 8, the dermal fibroblast after In vitro culture 4-6 week is in the physiological water of 9,000,000 every milliliter-9,800,000 ratio adding 0.9%.Make cell suspension.
The mechanism of action of injection: when from the fibroblast re-injection of body in my wrinkle substrate skin corium, the fibroblast that survives can secrete a large amount of collagen protein, plays the effect of reduce wrinkle.
Three. product detects
1. virus detects: after around skin histology is cultivated, get the check that culture fluid that 1ml contains cell send Beijing infectious hospital to carry out HIV and hepatitis virus, determine that this is virus-free cell.
2. immunoreation test: around the 0.1ml cell suspending liquid is carried out skin test (with penicillin skin test method) to the operation receiveing person, to guarantee not having immunoreation.
3. finished product detection
1) outward appearance: this product is the pale suspension.
2) sterility test: undertaken by " Chinese biological goods rules " (version in 2000) general rule " biological product sterility test rules " A/B item, the result meets aseptic requirement.
The storage of product and transportation should be carried out in 2 ℃-10 ℃, shady and cool, dry, the environment that cleans, avoid sunlight direct projection, non-corrosiveness gas, no weight.
Clinical report
One, case is selected: the age is at the right side of fifty (comprising 50 years old), wrinkle be forehead, near the eyes, perioral aging lines, cicatrix is acne, wound cicatrix (magnitude range no requirement (NR)).
Two, therapeutic process
1. check before patient's art: blood, urine, just routine, electrocardiogram, hepatic and renal function, HIV, HbsAg.
With the injection site with 70% alcohol disinfecting, the lignocaine of local injection 1%, perhaps epidermis is with 2.5% lignocaine and the anesthesia of 2.5% Astral512 ointment.
3. the syringe that 1.5ml cell suspension is sucked 3ml is standby.
4. get No. 4.5 long syringe needles of 2.2cm, adopt multiple spot inclined-plane (the angle 20-45 of syringe needle and skin °) that cell suspension is injected into the upper strata and the middle level of skin corium, the 10-15 point is chosen in each injection site, injects cell suspension 0.05-0.1ml at every.Should it be turned white the tension of injection site epidermis during injection, should make the injection site leave the disperse space during injection.Wrinkle or cicatrix portion are all injected, injected the back and used ice bag external application 2 hours.
Injection site can have sensations such as mild swelling, numbness in the postoperative 24 hours, does not generally have other part and general reaction.
The course of treatment: general wrinkle injection 2 times, big wrinkle or cicatrix injection three times, per injection is 2-4 week at interval.
Four, clinical follow, nurse
1. after the operation, apply injection place 2 hours with ice bag.
2. observe the situation of part and whole body.
3. oral vitamin C every days two (each 200mg, every day is 400mg altogether), perhaps the part is put on the skin and is contained ascorbic facial cream, about 6 months.
4. postoperative was prevented tanning by the sun and the careful zest cosmetics of using in 3 days.
Five, efficacy analysis
Analysis of cases through 65 examples, wherein the case of a shot has 42 examples, double injection 15 examples are arranged, be 6 months the observing time between wherein injecting for the first time and for the second time, 58 examples of patient's approbation (satisfaction rate is 89.2%) as a result, its wrinkle, cicatrix obviously disappear, and the dermal matrix situation improves.
Wherein there are 5 routine therapists to observe so far 3 years, the complete obiteration of its wrinkle, cicatrix.Respond well.
Though described the present invention by embodiment, those of ordinary skills know, without departing from the spirit and substance in the present invention, just can make the present invention that many distortion and variation are arranged, and scope of the present invention is limited to the appended claims.
Claims (7)
1. a preparation method that is used for the injection of Wrinkle-and scar-removing is characterized in that, comprising:
A. take off skin histology on one's body from the patient;
B. the skin histology that is taken off is put into incubator and carry out amplification cultivation;
C. the fibroblast of the amplification cultivation ratio in 9,000,000 every milliliter-9,800,000 is added in the standard liquid injection, make cell suspension.
2. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1 is characterized in that, described steps A further comprises:
Iodine piece disinfectant solution with 5% carries out local surfaces with 2% lignocaine then and anaesthetizes behind ear or the inboard sterilization of arm.Take off with the holostrome skin of cutisector with the 3-4 square millimeter, put into tissue and preserve liquid, wound site is sewed up, bandage covers.
3. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1 is characterized in that, described step B further comprises:
B1. incubator digestion 10 minutes is put in the piece of tissue and the tissue fluid that obtain;
B2. remove epidermis and subepidermal fat;
B3. piece of tissue is cut into some, and with some block organizations of being cut into being mutually that six well culture plates are put at 0.5 centimetre interval;
B4. six well culture plates are carefully put into incubator, add 0.5 milliliter of culture fluid gently along the tank wall of incubator after 4 hours, add 1 milliliter of culture fluid after 24 days again;
B5. piece of tissue is gone down to posterity and cultivate 4-6 week.
4. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1 is characterized in that, described step B5 further comprises: changed liquid once every 2-3 days, go down to posterity after 50%-70% compiles cell when cell reaches.
5. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1 is characterized in that the size of plurality of small blocks is about the 1-2 square millimeter described in the described step B3.
6. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1, the described standard liquid injection among the described step C is the normal saline solution of concentration 0.9%.
7. the preparation method that is used for the injection of Wrinkle-and scar-removing according to claim 1, standard liquid injection described in the described step C is the glucose injection of concentration 5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2004100888029A CN1768856A (en) | 2004-11-04 | 2004-11-04 | Method for preparing injection for removing wrinkles and scars |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2004100888029A CN1768856A (en) | 2004-11-04 | 2004-11-04 | Method for preparing injection for removing wrinkles and scars |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1768856A true CN1768856A (en) | 2006-05-10 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2004100888029A Pending CN1768856A (en) | 2004-11-04 | 2004-11-04 | Method for preparing injection for removing wrinkles and scars |
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| Country | Link |
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| CN (1) | CN1768856A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109010247A (en) * | 2018-09-20 | 2018-12-18 | 浙江卫未生物医药科技有限公司 | A kind of application and preparation method of the excretion body freeze-dried powder in autologous fibroblasts source |
| CN113117148A (en) * | 2018-08-15 | 2021-07-16 | 白晋 | Botulinum-binding autologous collagen injection |
-
2004
- 2004-11-04 CN CNA2004100888029A patent/CN1768856A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113117148A (en) * | 2018-08-15 | 2021-07-16 | 白晋 | Botulinum-binding autologous collagen injection |
| CN109010247A (en) * | 2018-09-20 | 2018-12-18 | 浙江卫未生物医药科技有限公司 | A kind of application and preparation method of the excretion body freeze-dried powder in autologous fibroblasts source |
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