CN1206963C

CN1206963C - Injection for eliminating wrinkles and scar and its prepn

Info

Publication number: CN1206963C
Application number: CNB03155833XA
Authority: CN
Inventors: 韩斌
Original assignee: Beijing Yiling Bioengineering Co Ltd
Current assignee: Lv Weiguang
Priority date: 2003-08-25
Filing date: 2003-08-25
Publication date: 2005-06-22
Anticipated expiration: 2023-08-25
Also published as: CN1486702A

Abstract

The present invention relates to an injection for eliminating wrinkles and scars and a preparation method thereof, which belongs to the field of tissue engineering. The injection is composed of ten million to fourteen million corium mechanocytes and 2 to 4 ml of glucose injection with the concentration of 5%. The preparation method comprises the following steps: corium mechanocytes which come from patients are in vitro expanded and cultured, and made into suspension according to that 2 to 4 ml of glucose injection with the concentration of 5% is added to every ten million to fourteen million corium mechanocytes. The present invention has the advantages that used biological materials are taken from patients and has the advantages of no rejection reaction or side effect, conspicuous beauty treatment curative effect, natural expression after being used by people to be treated. The effect is lasting and can be maintained for several years.

Description

Injection of a kind of Wrinkle-and scar-removing and preparation method thereof

Technical field

The present invention relates to injection of a kind of Wrinkle-and scar-removing and preparation method thereof, belong to field of tissue engineering technology.

Background technology

People adopt the collagen protein of injection cattle to treat wrinkle and cicatrix before 20 years, but it anaphylaxis can occur, can be organized simultaneously faster to absorb the time that can only keep some months.In order to overcome this anaphylactoid shortcoming, some hospital adopts and becomes collagen protein to inject from the body leather, and the result is that preparation process needs a large amount of skin, and still can be organized very fast absorption after the injection.Many in recent years people adopt creotoxin to inject and treat wrinkle and cicatrix, and its result also can only keep the relatively shorter time, and the expression utmost point of treatment back therapist is unnatural.

Summary of the invention

In order to overcome the weak point of these products, the purpose of this invention is to provide a kind of efficient, permanent, injection of the Wrinkle-and scar-removing of back nature more.

The autologous skin that this injection has adopted the people is isolated dermal fibroblast through digestion, carries out amplification in vitro, is then injected into the wrinkle or the cicatrix position of therapist, and these cells produce collagen tissue in its injection site, smoothes away wrinkles and fills and leads up cicatrix.

Another object of the present invention provides the preparation method of this injection.

For achieving the above object, the present invention is by the following technical solutions:

A kind of injection of Wrinkle-and scar-removing is made up of the glucose injection of 1,000 ten thousand-4,000 ten thousand dermal fibroblasts and 2-4ml concentration 5%.

Described dermal fibroblast is to obtain through amplification in vitro, cultivation deriving from the dermal fibroblast of patient from body.

A kind of preparation method of injection of Wrinkle-and scar-removing, be to derive from the patient to cultivate at amplification in vitro from the dermal fibroblast of body, add the glucose injection of 2-4ml concentration 5% again according to per 1,000 ten thousand-4,000 ten thousand dermal fibroblasts, make suspension.

The present invention a kind ofly is expelled to the Wrinkle-and scar-removing product that wrinkle, cicatrix position are repaired with patient's autologous skin cell external once more through propagation, after cultivating, collagen protein makes rill and cicatrix is long puts down thereby this product produces at these positions, smoothing away wrinkles and cicatrix can improve simultaneously the environment of local skin again, playing the effect of young U.S. face.

Advantage of the present invention: the biomaterial that the present invention uses is taken from the patient from body, and no rejection has no side effect, therapist with after improve looks evident in efficacyly, look natural, and the effect effect is lasting, can keep the several years.

The invention will be further described for the following specific embodiment.

The specific embodiment

The preparation of the injection of embodiment 1, Wrinkle-and scar-removing

One. from the extraction of body healthy cell

With behind 70% the alcohol disinfecting ear or arm inboard, use then 10% lignocaine and 100,000/ epinephrine carry out local surfaces anesthesia.Take off with the epidermis and the dermal tissue of cutisector with 4 square millimeters, put into tissue and preserve in the liquid (DMEM and 10% hyclone, U.S. Hyclone company), wound site is made a pin sew up, adhesive bandage covers.

Two. the preparation of cell suspension

1. tissue digestion (Sanya CO2 incubator): the skin histology piece is positioned over to add 2ml concentration in the 35mm culture dish be pancreatin/EDTA liquid of 0.05%, 37 ℃, 10 minutes; The DMEM+10%FBS culture fluid stops.

2. cell separation: postdigestive piece of tissue is shredded and breaks up with suction pipe.

3. centrifugal: the cell harvesting after dispelling to centrifuge tube, Backman company temperature control centrifuge, 25 ℃, 1000 commentaries on classics/min, centrifugal 5 minutes, abandoning supernatant.

4. primitive cell culture: add the low serum true chrotoplast culture fluid (Sigma company, the U.S.) of 1ml and blow outstandingly in centrifuge tube, 1 * 35mm culture dish is gone in inoculation; Sanya CO ₂Incubator, 37 ℃, 5% CO ₂, cultivate.

5. passage cell is cultivated: changed liquid 1 time every 2-3 days, go down to posterity after 50-70% compiles cell when cell reaches.

6. preparation injection: the dermal fibroblast after subculture in vitro separately to the 4-6 week, adding 2-4ml concentration by per 1,000 ten thousand-4,000 ten thousand fibroblasts is 5% glucose injection, is mixed with suspension.

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 " (2000 editions) 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.

Embodiment 2, clinical report

One. case standard (case load 28 examples)

1. include standard in: the age (comprised 55 years old) below 55 years old; The wrinkle person be forehead, near the eyes, perioral aging lines; The cicatrix person is acne, wound cicatrix (magnitude range no requirement (NR)).

2. exclusion standard: the age is patient more than 55 years old; Autoimmune disease is arranged; The chronic skin patient is arranged; The infection patient is arranged.

Two. prepare injection: the method according to embodiment 1 is carried out.

Three. treatment

1. check before patient's art: hematuria is routine, electrocardiogram, hepatic and renal function, HIV, HbsAg just.

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. standby in the syringe with 1.5ml cell suspension suction 3ml.

4. get No. 4.5 long syringe needles of 2.2cm, adopt multiple spot inclined-plane (20 °-45 ° of the angles of syringe needle and skin) that cell suspension is injected into the upper strata and the middle level of skin corium, the 15-20 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 has 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. secondary oral vitamin C every day (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

Through to the analysis of cases of 28 examples, wherein the case of a shot has 20 examples, double injection 8 examples are arranged, 6 months observing times, the result has 24 examples (its satisfaction rate is 85.7%) for my approbation of patient, and its wrinkle, cicatrix obviously disappear, and the dermal matrix situation obviously improves.

The person observed 2 years so far wherein to have the individualized treatment, the complete obiteration of its wrinkle, cicatrix, skin smooth and high resilience.

Claims

1, a kind of wrinkle removal is removed the injection of scar, it is characterized in that: this injection is made up of the glucose injection of 1,000 ten thousand-4,000 ten thousand dermal fibroblasts and 2-4ml concentration 5%, dermal fibroblast wherein be origin come from the patient from the dermal fibroblast of body through amplification in vitro, cultivate and obtain.

2, the described wrinkle removal of a kind of claim 1 is removed the preparation method of the injection of scar, it is characterized in that: this method be with derive from the patient from the dermal fibroblast of body in amplification in vitro and cultivation, add the glucose injection of 2-4ml concentration 5% again according to per 1,000 ten thousand-4,000 ten thousand dermal fibroblasts, make suspension.