CN114344454A - Preparation method of autologous fat collagen injection - Google Patents
Preparation method of autologous fat collagen injection Download PDFInfo
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- CN114344454A CN114344454A CN202210071217.6A CN202210071217A CN114344454A CN 114344454 A CN114344454 A CN 114344454A CN 202210071217 A CN202210071217 A CN 202210071217A CN 114344454 A CN114344454 A CN 114344454A
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method of an autologous fat collagen injection, which comprises the following steps: s1, culturing the autologous fat at constant temperature; s2, cutting and scattering the autologous fat processed by the S1, and centrifuging to remove oil; s3, blowing and suspending the fibroblast of S2, and culturing at constant temperature to make the fibroblast secrete collagen; s4, placing the venous blood sample in a test tube containing anticoagulant, centrifuging the supernatant to obtain plasma rich in platelet and growth factor; s5, mixing the fibroblasts, the collagen and the plasma in the S4 uniformly, and obtaining the body fat collagen injection. According to the invention, 90% of useless oil drops in autologous fat are removed, and a centrifugal technology is combined to continuously remove components with poor support and low survivability, so that the collagen with good support and high survivability is finally obtained; mixing collagen with plasma rich in platelet and growth factor, the collagen can absorb nutrition in PRP and largely replicate and regenerate, so that the depressed part is filled, and the skin becomes full.
Description
Technical Field
The invention belongs to the field of medical cosmetology, and particularly relates to a preparation method of autologous fat collagen.
Background
The wrinkles refer to small fine lines and wrinkles formed by the skin affected by external environment, which form free radicals, the free radicals destroy collagen and active substances in normal cell membrane tissues and oxidize cells. The occurrence of wrinkles is attributed to 4 causes: natural aging, gravity effect, and ultraviolet irradiation in sunlight can cause skin photoaging and photodamage and excessive contraction of facial expression muscles. Among them, the wrinkles caused by the excessive contraction of facial expression muscles are called dynamic wrinkles and mainly appear as fishtail lines on the lateral orbit, forehead lines and interphalangeal lines, nasolabial sulcus lines and superior labial lines and platysma lines. There are many methods for removing wrinkles, and topical application of various wrinkle-removing creams, wrinkle-resisting creams, and sunscreen creams can only improve appearance, and it does not have great effect on wrinkle removal.
Collagen is the main component of extracellular matrix, accounts for about 85% of the solid content of collagen fiber, accounts for 25% -30% of the total amount of protein in the animal body, and type I collagen is mainly distributed in tissues such as skin, tendon and the like, is also the protein with the largest content of aquatic product processing waste (skin, bone and scale), accounts for about 80% -90% of the total content of collagen, and is most widely applied in medicine. The lower layer of human skin also has densely arranged collagen, the thickness and the components of the skin can change along with the increase of age, the skin of the old is easy to be injured and is not easy to heal, which is probably caused by the loss of fibroblasts of the upper layer of skin, if the growth of the cells can be stimulated, the elasticity of the skin can be recovered, and the formation of hair follicles can also be stimulated, so that scars can be reduced. Wrinkles are removed by means of autologous collagen injection, and anaphylactic reaction of medical collagen can be remarkably reduced.
However, the enriched type I collagen is not contained in autologous adipose tissue, and besides the difficulty of obtaining it and the special requirements of liposuction equipment, it is necessary for liposuction surgeons to master the technique for preparing autologous adipose collagen. In addition, simple collagen injection is easily degraded by the body, and cannot fundamentally solve the problem of wrinkles. Therefore, how to solve the problems that the acquisition difficulty of the type I collagen is high and the collagen is easily degraded by the organism has important significance for the beauty industry.
Disclosure of Invention
In view of the above, the present invention provides a method for preparing an autologous fat collagen injection.
In order to achieve the purpose, the invention adopts the following technical scheme:
the preparation method of the autologous fat collagen injection comprises the following steps:
s1, placing the autologous fat in EDTA solution containing trypsin, culturing at the constant temperature of 37 ℃ for 10-20min, and adding EDTA solution containing fetal calf serum into the culture solution to terminate the culture;
s2, cutting and scattering the autologous fat digested in the step S1, and centrifuging to remove 90% of oil drops to obtain fibroblasts;
s3, the fibroblasts obtained in S2 were suspended in a low serum cell culture solution and then inoculated into the low serum cell culture solution at 37 ℃ with 5% CO2Culturing at constant temperature to make fibroblasts secrete collagen;
s4, placing the venous blood sample into a test tube containing anticoagulant, centrifuging at 600rpm, sucking the supernatant and the liquid with the height of 3mm below the interface by using a suction tube, transferring the supernatant and the liquid into a new test tube, and centrifuging to obtain the plasma rich in platelets or growth factors in the middle layer of the test tube;
s5, after the collagen in S3 is cultured, taking out fibroblasts and collagen from the low serum cell culture solution to obtain fibroblasts and collagen; adding the plasma rich in platelets or growth factors obtained from S4 into collagen and fibroblasts, and uniformly mixing to obtain the body fat collagen injection.
Preferably, the mass fraction of trypsin in S1 is 0.05%; the mass fraction of FBS is 10%.
Preferably, the culture time of the fibroblast in S3 is 12-36 h.
Preferably, the centrifugation of the venous blood sample in S4 is for 10 min.
The method has the advantages that 90% of useless oil drops in the autologous fat are removed, and the centrifugal technology is combined to continuously remove components with poor support and low survivability in the autologous fat, so that the collagen with good support and high survivability is finally obtained; and mixing collagen with plasma (PRP) rich in platelet and growth factor, the collagen can absorb nutrition in PRP and largely copy and regenerate collagen, thereby filling the depressed part and making skin plump.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The preparation method of the autologous fat collagen injection comprises the following steps:
s1, adding 2mL of EDTA solution containing trypsin into autologous fat, culturing at the constant temperature of 37 ℃ for 10min, and then adding EDTA solution containing fetal calf serum into the culture solution to terminate the culture so as to digest the autologous fat;
wherein the mass fraction of the trypsin is 0.05%; the mass fraction of FBS (fetal bovine serum) is 10 percent;
s2, cutting the digested autologous fat in the step S1 into pieces, scattering the cut autologous fat by a suction pipe, centrifuging the scattered fat for 5min under the centrifugal conditions of 25 ℃ and 1000r/min, and discarding 90% of useless oil drops in 90 autologous fat to obtain fibroblasts;
s3, adding 1mL of low serum dermal cell culture solution (sienna corporation, usa) to the fibroblasts in step S2, and blowing the fibroblasts; then inoculating the blown and suspended fibroblasts into a low-serum dermatin cell culture solution at 37 ℃ and 5% CO2Culturing at constant temperature for 12-36h to make fibroblasts secrete collagen;
s4, placing a venous blood sample (10 mL) in a test tube containing anticoagulant, centrifuging at 600rpm for 10min, sucking the supernatant and liquid with the height of 3mm below an interface by using a suction tube, transferring the supernatant and the liquid to a new test tube (without anticoagulant), centrifuging at 600rpm for 10min, wherein the centrifuged plasma is three layers, and the middle layer is plasma rich in platelets and growth factors (the growth factors comprise PDGF, VEGF, IGF-1, EGF and TGF-beta), namely PRP;
s5, after the collagen in S3 is cultured, taking out fibroblasts and collagen from the low serum cell culture solution to obtain fibroblasts and collagen; adding the plasma rich in platelets or growth factors obtained from S4 into collagen and fibroblasts, and uniformly mixing to obtain the body fat collagen injection.
The invention removes 90% useless oil drops in the autologous fat during preparation, combines the centrifugal technology to continuously remove components with poor support and low survivability in the autologous fat, obtains 3% of fibroblasts with good support and high survivability in the autologous fat, and obtains collagen with good support through enlarged culture. The injection of the compound into aging-damaged subcutaneous cells can enable new muscle cells to rapidly replace aged muscle cells and enable the aged muscle cells to revive the vitality;
the invention adds PRP rich in juvenile stem cells, PLT (platelet) and growth factors (PDGF, VEGF, IGF-1, EGF and TGF-beta) into collagen, the collagen can absorb a large amount of nutrition in the PRP and copy and regenerate the collagen in a large amount, thereby filling the sunken part and ensuring that the skin becomes full.
According to the invention, collagen and PRP are mixed together, and the growth factor in PRP can enhance the phagocytosis and decomposition capacity of the cytochrome cells, provide energy power for the generation of transport catabolism, and repair normal skin tissues damaged by pigmentation, thereby achieving the effects of whitening, lightening spots and tendering skin.
Claims (4)
1. A preparation method of an autologous fat collagen injection is characterized by comprising the following steps:
s1, placing the autologous fat in EDTA solution containing trypsin, culturing at the constant temperature of 37 ℃ for 10-20min, and adding EDTA solution containing fetal calf serum into the culture solution to terminate the culture;
s2, cutting and scattering the autologous fat digested in the step S1, and centrifuging to remove 90% of oil drops to obtain fibroblasts;
s3, the fibroblasts obtained in S2 were suspended in a low serum cell culture solution and then inoculated into the low serum cell culture solution at 37 ℃ with 5% CO2Culturing at constant temperature to make fibroblasts secrete collagen;
s4, placing the venous blood sample into a test tube containing anticoagulant, centrifuging at 600rpm, sucking the supernatant and the liquid with the height of 3mm below the interface by using a suction tube, transferring the supernatant and the liquid into a new test tube, and centrifuging to obtain the plasma rich in platelets or growth factors in the middle layer of the test tube;
s5, after the collagen in S3 is cultured, taking out fibroblasts and collagen from the low serum cell culture solution to obtain fibroblasts and collagen; adding the plasma rich in platelets or growth factors obtained from S4 into collagen and fibroblasts, and uniformly mixing to obtain the body fat collagen injection.
2. The method for preparing an autologous fat collagen injection according to claim 1, wherein the method comprises the steps of: the mass fraction of trypsin used in the S1 is 0.05%; the mass fraction of FBS is 10%.
3. The method for preparing an autologous fat collagen injection according to claim 1, wherein the method comprises the steps of: the culture time of the fibroblast in the S3 is 12-36 h.
4. The method for preparing an autologous fat collagen injection according to claim 1, wherein the method comprises the steps of: centrifugation of the venous blood sample in S4 for 10 min.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210071217.6A CN114344454A (en) | 2022-01-21 | 2022-01-21 | Preparation method of autologous fat collagen injection |
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| CN202210071217.6A CN114344454A (en) | 2022-01-21 | 2022-01-21 | Preparation method of autologous fat collagen injection |
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| CN114344454A true CN114344454A (en) | 2022-04-15 |
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| CN202210071217.6A Pending CN114344454A (en) | 2022-01-21 | 2022-01-21 | Preparation method of autologous fat collagen injection |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116407559A (en) * | 2023-05-25 | 2023-07-11 | 云南康旭生物科技有限公司 | Preparation for treating premature ovarian failure and preparation method thereof |
| CN116492509A (en) * | 2023-06-13 | 2023-07-28 | 哈尔滨悦之美芳华医疗美容门诊有限公司 | Filler for medical shaping and preparation method thereof |
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