CN109106983B - Fat transplantation material and preparation method and application thereof - Google Patents
Fat transplantation material and preparation method and application thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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Abstract
The invention discloses a fat transplantation material and a preparation method and application thereof, wherein the preparation method comprises the following steps: (a) selecting fat particles, and centrifugally collecting middle-layer adipose tissues; (b) stirring the adipose tissues by using a stirring rod, collecting the fibrous tissues wound on the stirring rod, and repeating the stirring and collecting operations until no fibrous tissues are wound after the stirring rod is stirred; (c) mixing the collected fibrous tissues with physiological saline, stirring and collecting precipitates; (d) removing water from the collected precipitate, and crushing into paste to obtain fat transplantation material; the preparation method is simple to operate, fast and efficient, does not need to use an exogenous reagent, avoids cross infection and improves the yield of the fat transplantation material.
Description
Technical Field
The invention relates to the technical field of cosmetic filling materials, in particular to a fat transplantation material and a preparation method and application thereof.
Background
In recent years, with the increasing number of people seeking beauty, the operation of nose augmentation and chin augmentation is also increased year by year. The filling materials clinically applied to the nose augmentation chin operation are many, and in the artificial synthetic materials, solid materials comprise silicon rubber, expanded polytetrafluoroethylene and the like, and the materials are easy to generate complications such as rejection, postoperative infection and the like; the soft filler is collagen, hyaluronic acid and the like, and the materials are absorbable materials, so that repeated filling is needed, and the cost is increased. The autologous tissue comprises a dermis flap, autologous auricular cartilage, costal cartilage and the like, and the autologous tissue flap or cartilage transplantation brings secondary trauma and deformity of the donor area.
As a soft tissue filler, autologous fat particles are increasingly paid more attention by domestic and foreign plastic cosmetologists due to the advantages of abundant sources, easily available materials, simple operation, good filling appearance, no rejection reaction and the like. Because the effect is obvious, the injury is small, the cost is low, the beauty seeking person who fills the face with the granular fat increases year by year, but a certain absorption rate exists after the fat transplantation, so that part of beauty seeking persons need to fill for many times, particularly in the nasal back, the chin and other parts, the survival rate of the transplanted fat is low due to the factors of small local tissue amount, insufficient vascular network, high skin tension and the like, and the ideal effect is difficult to achieve.
At present, an adipose tissue stem cell gel is disclosed in the prior art, in the preparation process of the adipose tissue stem cell gel, fat particles need to be emulsified and then centrifuged, however, a great amount of adipose stem cells are damaged in the emulsifying process, so that the yield of the adipose tissue stem cell gel and the survival rate after transplantation are low; moreover, the adipose tissue stem cell gel contains a large amount of cell fragments which are easily absorbed by the gel, so that the adipose tissue stem cell gel has high absorption rate when being used for nose augmentation and chin augmentation; in addition, the oily cell contents released after the adipocytes are destroyed are not valuable and are discarded, resulting in waste of resources.
Disclosure of Invention
A first object of the present invention is to provide a method for preparing a fat transplantation material, which is simple in operation, rapid and efficient, does not require the use of an exogenous reagent, avoids cross-infection, and improves the yield of the fat transplantation material.
A second object of the present invention is to provide a fat graft material, which is obtained by the above method, has a high survival rate after transplantation and has a low absorption rate.
The third purpose of the invention is to provide an application of the fat transplantation material, in particular to an application of the fat transplantation material in the parts of nose augmentation, chin augmentation and the like, and to achieve higher survival rate.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
the first aspect of the present invention provides a method for preparing a fat transplantation material, comprising the steps of:
(a) selecting fat particles, and centrifugally collecting middle-layer adipose tissues;
(b) stirring the adipose tissues by using a stirring rod, collecting the fibrous tissues wound on the stirring rod, and repeating the stirring and collecting operations until no fibrous tissues are wound after the stirring rod is stirred;
(c) mixing the collected fibrous tissues with physiological saline, stirring and collecting precipitates;
(d) removing water from the collected precipitate, and crushing into paste to obtain the fat transplantation material.
Preferably, the preparation process of the fat transplantation material is carried out under aseptic conditions, and the adopted reagents and instruments are sterilized in advance.
The preparation method can fully separate fibrous tissues in the fat particles, avoid the damage of fat stem cells and improve the yield of the fat transplantation material; the preparation method does not need to use an exogenous reagent, avoids cross infection, and has the advantages of simple operation, high speed and high efficiency and easy production.
Further, the preparation method comprises the steps of placing the obtained fat transplantation material into two syringes connected by a nano fat converter, pushing and pulling for 2-3 times, removing oil, and shearing to obtain the purified fat transplantation material. The fat transplantation material is purified, so that the collagen content is further improved, and the plasticity and the supporting strength after the nose is humped and the chin is raised are improved.
The fat can be fully utilized in the invention, the obtained fat transplantation material is used for filling special parts such as the nasal back, the chin and the like, and the residual granular fat after extracting the lipocollagen can be used for filling other parts of the face or the chest and the like.
The origin of the fat particles in the present invention is not strictly limited, and preferably, the fat particles in the technical scheme of the present application are selected from abdominal fat; by selecting specific abdominal fat, the content of collagen fibers in the prepared fat transplantation material can be improved, the plasticity of the fat transplantation material is improved, and the fat transplantation material is not easy to deform.
Further, in the step (a), the centrifugal rotating speed is 900-; the centrifugation time is 2-4 min. The separation of fibrous tissues can be promoted by the centrifugal arrangement, however, the separation of the fibrous tissues is not easy to realize due to the low centrifugal rotating speed, the fibrous tissues are broken due to high centrifugal rotating speed, the subsequent extraction of the fibrous tissues is not facilitated, and the yield of the fat transplantation material is directly influenced; the invention can improve the sufficient separation of fibrous tissues by specifically limiting the centrifugal conditions, and is beneficial to improving the yield of the prepared fat transplantation material.
Further, in the step (b), the type of the stirring rod is not strictly limited, and for example, the stirring rod may be a conventional stirring rod in the art, and may also be a grease injection tube or a hemostatic forceps; preferably an 18G grease gun.
Further, in the step (c), the volume ratio of the physiological saline to the fat particles is 1 to (2-4). The fat in the fibrous tissue can be removed by the addition of the normal saline and the limitation of the addition amount, the residual fat is avoided, and the fat after transplantation is absorbed by the tissue of the body, so that the absorption rate of the fat transplantation material is higher.
Further, in the step (d), the step of removing water is to absorb water on the precipitate by using gauze; the crushing is carried out by shearing with scissors.
The present invention provides, in a second aspect, a fat graft material produced by the above production method. The fat transplantation material has high survival rate after transplantation and low absorption rate.
Further, the fat transplantation material comprises stromal vascular segment cells, adipose stem cells and collagen. Research shows that the survival rate of the fat transplantation material after transplantation can be better improved through the combined action of stromal vascular segment cells and fat stem cells; in addition, a large amount of collagen fibers contained in the fat transplantation material have high density, high viscosity, strong supporting force and strong shaping capacity, and are not easy to deform and swell.
Preferably, the stromal vascular fraction has a cell content of (6.2-9.5). times.106Per ml; the content of adipose-derived stem cells is (1.4-3.1) x 106Per ml; the collagen content is 74% -98%. The invention can better improve the survival rate of the transplanted fat transplantation material by limiting the content of each component in the fat transplantation material.
Preferably, the collagen is type i collagen.
In a third aspect, the present invention provides a use of a fat graft material for augmentation of nose, augmentation of chin, augmentation of nasolabial fold, augmentation of lacrimal fold, augmentation of depressed scar or facial rejuvenation.
Further, the fat transplantation material is applied to hump nose or chin. The fat transplantation material is applied to the nasal back or the chin and other parts with less tissue, less vascular network, high skin tension and the like, and achieves the aim that the transplanted fat transplantation material has higher survival rate.
Compared with the prior art, the invention has the beneficial effects that:
(1) the preparation method of the fat transplantation material is simple to operate, fast and efficient, does not need to use an exogenous reagent, avoids cross infection and improves the yield of the fat transplantation material.
(2) The fat transplantation material can better improve the survival rate of the transplanted fat transplantation material through the combined action of the stromal vascular segment cells and the adipose-derived stem cells with specific content.
(3) The fat transplantation material contains a large amount of collagen fibers, has high density and high viscosity, has strong supporting force, has strong shaping capacity, and is not easy to deform and overstaffed.
(4) The fat particles adopted by the invention are selected from the abdominal fat, so that the content of collagen fibers in the prepared fat transplantation material can be improved, the plasticity of the fat transplantation material is improved, and the fat transplantation material is not easy to deform.
(5) The fat transplantation material can be applied to the parts of hump nose, hump chin and the like, and achieves higher survival rate of the transplanted fat transplantation material.
(6) The preparation method does not damage fat particles, and after the fibrous tissues are extracted, the remaining fat particles can be used for filling the face or the chest, so that the resources are fully utilized.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by manufacturers, and are all conventional products available on the market.
Example 1
This embodiment is a method for preparing a fat transplantation material, including the steps of:
(a) selecting 100ml of abdominal fat particles, centrifuging for 2min at the rotating speed of 1000r/min, and then collecting the middle-layer adipose tissues to a sterile medicine placing bowl;
(b) stirring the adipose tissues by adopting an 18G grease injection pipe, collecting the fibrous tissues wound on the 18G grease injection pipe into an aseptic bent disc, and repeating the stirring and collecting operations until no fibrous tissues are wound after the 18G grease injection pipe is stirred;
(c) adding 50ml of sterile normal saline into a sterile bent disc, stirring, and collecting a precipitate;
(d) placing the collected precipitate on sterile gauze, removing water, placing in a sterile small cup, and shearing fibrous tissue into paste with scissors;
(e) placing the pasty precipitate into two 10ml syringes connected by nanometer fat converter, pushing and pulling for 2 times, placing the treated precipitate on gauze to remove oil, taking down fibrous tissue with forceps, and cutting to obtain fat transplantation material.
Weighing the fat transplantation material prepared by the method, and calculating the yield of the fat transplantation material to be 11.8%; and detecting the obtained fat transplantation material by defatting, collagenase digesting, centrifuging, cell counting and flow cytometry analysis, wherein the cell content of the stromal vascular fraction is 9.5 × 106Per ml; the content of adipose-derived stem cells is 2.1 × 106Per ml; the collagen content was 91%.
Example 2
This embodiment is a method for preparing a fat transplantation material, including the steps of:
(a) selecting 200ml of abdominal fat particles, centrifuging for 4min at the rotating speed of 1000g/min, and then collecting the middle-layer adipose tissues to a sterile medicine placing bowl;
(b) stirring adipose tissues by using a hemostatic forceps, collecting fibrous tissues wound on the hemostatic forceps into an aseptic bent disc, and repeating the stirring and collecting operations until no fibrous tissues are wound after the hemostatic forceps are stirred;
(c) adding 50ml of sterile normal saline into a sterile bent disc, stirring, and collecting a precipitate;
(d) placing the collected precipitate on sterile gauze, removing water, placing in a sterile small cup, and shearing fibrous tissue into paste with scissors;
(e) placing the pasty precipitate into two 10ml syringes connected by nanometer fat converter, pushing and pulling 3 times, placing the treated precipitate on gauze to remove oil, taking down fibrous tissue with forceps, and cutting to obtain fat transplantation material.
Weighing the fat transplantation material prepared by the method, and calculating the yield of the fat transplantation material to be 11.5%; and the fat transplantation material prepared above was examined by the examination method of example 1, in which the stromal vascular fraction cell content was 6.2X 106Per ml; the content of adipose-derived stem cells is 1.4 × 106Per ml; the collagen content was 92%.
Example 3
This embodiment is a method for preparing a fat transplantation material, including the steps of:
(a) selecting 150ml of abdominal fat particles, centrifuging for 3min at the rotating speed of 1000g/min, and then collecting the middle-layer adipose tissues to a sterile medicine placing bowl;
(b) stirring the adipose tissues by adopting an 18G grease injection pipe, collecting the fibrous tissues wound on the 18G grease injection pipe into an aseptic bent disc, and repeating the stirring and collecting operations until no fibrous tissues are wound after the 18G grease injection pipe is stirred;
(c) adding 50ml of sterile normal saline into a sterile bent disc, stirring, and collecting a precipitate;
(d) placing the collected precipitate on sterile gauze, removing water, placing in a sterile small cup, and shearing fibrous tissue into paste with scissors;
(e) placing the pasty precipitate into two 10ml syringes connected by nanometer fat converter, pushing and pulling for 2 times, placing the treated precipitate on gauze to remove oil, taking down fibrous tissue with forceps, and cutting to obtain fat transplantation material.
Weighing the fat transplantation material prepared by the method, and calculating the yield of the fat transplantation material to be 12.3%; and the fat transplantation material prepared above was examined by the examination method of example 1, in which the stromal vascular fraction cell content was 7.7X 106Per ml; the content of adipose-derived stem cells is 1.5 × 106Per ml; the collagen content was 95%.
Example 4
This example is a method for preparing a fat graft material, which is substantially the same as that of example 3 except that abdominal fat particles are replaced with leg fat particles in step (a).
Weighing the fat transplantation material prepared by the method, and calculating the yield of the fat transplantation material to be 9.4%; and the fat transplantation material prepared above was examined by the examination method of example 1, in which the stromal vascular fraction cell content was 6.4X 106Per ml; the content of adipose-derived stem cells is 3.1 × 106Per ml; the collagen content was 94%.
Comparative example 1
The present comparative example is an adipose-derived transplantation material containing highly concentrated adipose-derived stem cells and extracellular matrix, which was prepared according to the preparation method of example 1 of chinese patent application publication No. CN 106178134a, and adipose particles were selected from abdominal fat.
Weighing the fat transplantation material prepared by the method, and calculating the yield of the fat transplantation material to be 10.3%; and the fat transplantation material prepared above was examined by the examination method of example 1, in which the stromal vascular fraction cell content was 3.4X 106Per ml; the content of adipose-derived stem cells is 1.5 × 106Per ml; the collagen content was 51%.
Examples of the experiments
Experimental materials:
selecting 12 healthy nude mice with the body weight of 20-25g and the age of 5-7 weeks;
selecting abdominal and leg adipose tissues of the same patient, and preparing fat transplantation materials according to the preparation methods of the above examples 2, 3 and 4 and the comparative example 1;
the experimental method comprises the following steps:
the experiments were divided into 4 groups: group a is the fat graft material prepared in example 2; group B is the fat graft material prepared in example 3; group C is the fat graft material prepared in example 4; group D was the fat graft material prepared in comparative example 1.
Respectively injecting the 4 components into four subcutaneous receptor areas at the waist and the back of an experimental nude mouse according to a randomization principle, wherein the injection amount of each component is 300 mg; then, the experimental nude mice were bred under the same conditions for 3 months.
The transplanted fat transplantation material was taken out of the nude mice after 3 months of rearing, the wet weight was measured immediately by using an electronic balance, and the average survival rate of 4 groups of injection components was calculated according to the survival rate ═ average wet weight/injection amount × 100%, and the specific results are shown in table 1; then observing the volume size, texture, color and distribution condition of blood vessels of the fat transplantation material, wherein the observation condition is shown in table 1;
TABLE 1
As can be seen from Table 1:
the fat transplantation material prepared by the preparation method of the embodiment 2 has the highest survival rate, and experimental data show that the contents of stromal vascular segment cells, adipose-derived stem cells and collagen have a synergistic effect on the survival rate of the fat transplantation material, and the fat transplantation material prepared by the embodiment 2 can be determined to have the lowest absorption rate by observing the volume and is obviously superior to the fat transplantation material prepared by the comparative example 1.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (9)
1. A method for preparing a fat transplantation material is characterized by comprising the following steps:
(a) selecting fat particles, and centrifugally collecting middle-layer adipose tissues;
(b) stirring the adipose tissues by using a stirring rod, collecting the fibrous tissues wound on the stirring rod, and repeating the stirring and collecting operations until no fibrous tissues are wound after the stirring rod is stirred;
(c) mixing the collected fibrous tissues with physiological saline, stirring and collecting precipitates;
(d) removing water from the collected precipitate, and crushing into paste to obtain the fat transplantation material.
2. The preparation method according to claim 1, further comprising placing the obtained fat graft material in two syringes connected by a nano fat converter, pushing and pulling 2-3 times, removing oil, and cutting to obtain a purified fat graft material.
3. The method of claim 1, wherein in step (a), the fat particles are selected from the group consisting of abdominal fat; the centrifugal rotating speed is 900-; the centrifugation time is 2-4 min.
4. The method of claim 1, wherein the volume ratio of the physiological saline to the fat particles is 1: 2-4.
5. A fat graft material produced by the production method according to any one of claims 1 to 4.
6. The fat graft material according to claim 5, wherein the fat graft material comprises stromal vascular fraction cells, adipose stem cells and collagen.
7. The fat graft material of claim 6, wherein the stromal vascular fraction cells compriseThe amount is (6.2-9.5). times.106Per ml; the content of adipose-derived stem cells is (1.4-3.1) x 106Per ml; the collagen content is 74% -98%.
8. The fat graft material according to claim 6, wherein said collagen is type I collagen.
9. The fat graft material according to any one of claims 5 to 8, which is used for nose augmentation, chin augmentation, nasolabial fold augmentation, lacrimal fold augmentation, depressed scar augmentation, or facial rejuvenation.
Priority Applications (1)
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