WO2008002064A1 - Composition de charge de tissus mous pour injection et son procédé de préparation - Google Patents
Composition de charge de tissus mous pour injection et son procédé de préparation Download PDFInfo
- Publication number
- WO2008002064A1 WO2008002064A1 PCT/KR2007/003100 KR2007003100W WO2008002064A1 WO 2008002064 A1 WO2008002064 A1 WO 2008002064A1 KR 2007003100 W KR2007003100 W KR 2007003100W WO 2008002064 A1 WO2008002064 A1 WO 2008002064A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- dermal
- cells
- injection
- autologous
- filler composition
- Prior art date
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- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- SOROIESOUPGGFO-UHFFFAOYSA-N diazolidinylurea Chemical compound OCNC(=O)N(CO)C1N(CO)C(=O)N(CO)C1=O SOROIESOUPGGFO-UHFFFAOYSA-N 0.000 description 1
- 229960001083 diazolidinylurea Drugs 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- WSDISUOETYTPRL-UHFFFAOYSA-N dmdm hydantoin Chemical compound CC1(C)N(CO)C(=O)N(CO)C1=O WSDISUOETYTPRL-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000001097 facial muscle Anatomy 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 210000000630 fibrocyte Anatomy 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- ZUVCYFMOHFTGDM-UHFFFAOYSA-N hexadecyl dihydrogen phosphate Chemical compound CCCCCCCCCCCCCCCCOP(O)(O)=O ZUVCYFMOHFTGDM-UHFFFAOYSA-N 0.000 description 1
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 1
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 238000007443 liposuction Methods 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000005155 neural progenitor cell Anatomy 0.000 description 1
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 201000005354 penicillin allergy Diseases 0.000 description 1
- 229960005323 phenoxyethanol Drugs 0.000 description 1
- 239000004584 polyacrylic acid Substances 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
- FRHNXUKHAUWMOQ-UHFFFAOYSA-M sodium;16-methylheptadecanoate Chemical compound [Na+].CC(C)CCCCCCCCCCCCCCC([O-])=O FRHNXUKHAUWMOQ-UHFFFAOYSA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- LADGBHLMCUINGV-UHFFFAOYSA-N tricaprin Chemical compound CCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCC)COC(=O)CCCCCCCCC LADGBHLMCUINGV-UHFFFAOYSA-N 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 210000000707 wrist Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0656—Adult fibroblasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/70—Polysaccharides
Definitions
- bovine collagen The preparation and application of bovine collagen have been disclosed in U.S. Patent Nos. 3,949,073, 4,424,208 and 4,488,911.
- the bovine collagen came into the market under the trade name Zyderm I , II and HI, wherein the content of collagen is in the range from 35 mg/ml to 65 mg/ml.
- Zyderm since an antibody to the bovine collagen is generated in 90% of the patients being treated with Zyderm (and 1 to 3% thereof shows clear allergic reactions), Zyderm does not provide satisfactory results.
- injectable glutaraldehyde cross-linked collagen has been developed and sold under the trade name Zyplast. Said protein does not cause any allergic reactions, but its viscosity is too strong to obtain good therapeutic effects (U.S. Patent Nos. 4,582,640 and 4,642,117).
- U.S. Patent Nos. 4,969,912 and 5,332,802 disclose a method of using autologous injectable human collagen for preventing an immune reaction due to the use of bovine collagen, which is available under the trade name AUTOLOGEN.
- Fibrel is based on a porcine collagen. Three items are combined to actually make up Fibrel, i.e., porcine gelatin, aminocaproic acid and plasma from the subject being treated. Fibrel shows the desired effects in some patients. However, most of the patients being treated are subject to swelling at the injection site and it is impossible to keep up its therapeutic effects for a long time.
- Artecoll is a complex of artificially synthesized particles and bovine collagen, which has been safely used in Canada, Mexico and Europe for several years. Artecoll exhibits prolonged therapeutic effects by providing autologous collagen through the stimulation of regional dermis. However, it has also problems in that the subject being treated can feel the presence of the particles and the swelling may occur at the treatment site.
- the effective ingredient of the injection disclosed in Chinese Patent No. 03155833.X is obtained by culturing the autologous skin sample in a medium partly supplemented with animal-derived serum.
- a medium partly supplemented with animal-derived serum there are several drawbacks to this method.
- the content of collagen in said injection which is responsible for the immediate onset of therapeutic effects, is too small, the effects for removing wrinkles and scars are late in terms of exertion, thereby taking 4 to 6 months to obtain satisfactory therapeutic effects.
- a subject being treated may worry about the safety due to the use of animal- derived serum as a component of the culture medium.
- compositions disclosed in said patents essentially consist of autologous muscle cells, autologous UMC or autologous keratinocytes, which are entirely different from a soft tissue filler composition comprising dermal fibroblast stem cells, dermal fibroblast transit amplifying cells and dermal fibroblasts described below.
- a method of preparing a soft tissue filler composition for injection which comprises the steps of:
- the autologous dermal biopsy used in Step 1) is prepared by disinfecting a target site on the skin of the patient being treated with alcohol, anesthetizing topically, excising epidermis and dermis in a size of 1 to 30 mm 2 therefrom, and then storing the same in a tissue stock solution.
- the skin tissue useful for the in vitro serum-free cultivation and proliferation of autologous dermal cells in this step may include all the skin, epidermis and dermis obtained from the back of the ear, eyebrows, the lower part of the eyes and other regions.
- prepared autologous dermal tissue sample is subjected to tissue digestion and cell isolation procedures.
- the basal medium useful for the present invention may be prepared by using conventional medium constitutions well known to one of ordinary skill in the art, e.g., ⁇ Cell Experimental Protocols>(2001, Science Press, Chief editor : D.L. Speycutter, Translation: Huang Peitang).
- the dermal cells isolated from the autologous dermal tissue are inoculated into the serum-free culture medium and cultured in an incubator at 37 °C under 5% CO 2 for 6 to 10 weeks. At this time, it is preferable to replace the medium with a fresh one at 3 day intervals.
- the autologous dermis-derived cell culture solution prepared above is centrifuged at a temperature ranging from 4 to 32 ° C at a speed ranging from 800 to 1,200 rpm/min to remove a supernatant, thereby recovering only an autologous dermis-derived cell pellet containing proliferated dermal fibroblast stem cells, dermal fibroblast transit amplifying cells, dermal fibroblasts and collagen secreted therefrom.
- the cultured and proliferated dermal fibroblast stem cells, dermal fibroblast transit amplifying cells and dermal fibroblasts and collagen secreted therefrom according to the present invention may be confirmed as follows:
- Alexa 594 Anti-Rabbit IgG Antibody, Molecular probe
- Alexa 488 Anti-Mouse IgG Antibody, Molecular probe
- the washed slide glass was observed with a fluorescent microscope (Olympus
- the dermis-derived cells cultured according to the method of the present invention exhibit a different expression pattern of cell markers from the adipose tissue-derived cells and display nestin (a marker for neural progenitor cells), they show immunological characteristics corresponding to neural progenitor-derived stem cells and not mesenchymal-derived stem cells (Toma et al., Stem Cells 23: 727-737, 2005; Fernandes et al., Nature 6: 1082-1093, 2004; and Toma et al., Nature 3: 778- 784, 2001).
- DAPI 4,',6-Diamidin-2-phenylindole dihydrochloride, SIGMA
- PBS fetal bovine serum
- DAPI 4,6-Diamidin-2-phenylindole dihydrochloride
- the slide glass was then soaked in the DAPI solution to stain the cells and washed three times with DPBS for 10 minutes.
- the washed slide glass was observed with a fluorescent microscope (Olympus IX 71) and recorded with a digital camera system.
- nestin and vimentin were expressed in the dermis-derived cells of the present invention.
- the cells expressing both of them were observed.
- the expression of vimentin was detected, but nestin was not expressed.
- Alexa 594 Anti-Rabbit IgG Antibody, Molecular probe
- Alexa 488 Anti-Mouse IgG Antibody, Molecular probe
- the dermis-derived cell culture material obtained by culturing in a serum-free medium according to the present invention shows signals for fibronectin and FSP-I known as a fibroblast marker protein as well as for nestin known as a neural progenitor-derived stem cell marker protein, which are clearly different from mesenchymal-derived stem cells such as adipose tissue-derived cells.
- the dermis-derived cells were cultured in a serum-free culture medium for 2 weeks according to the same method as described in Example 5.
- 0.25% trypsin/EDTA solution was added to the culture solution to detach the cells from the culture plate, followed by RNA extraction by using a RNA extraction kit (Purelink Micro to-midi, Invitrogen).
- a reaction solution was prepared by mixing thus extracted RNA, dNTP and oligo-dT 20 mer primer and adjusting its final volume to 10 ⁇ Jt.
- the reaction solution was incubated at 65 ° C for 5 minutes, followed by keeping at 0 ° C for a minute.
- 10 ⁇ i of a cDNA synthetic mixture (1OxRT buffer, 25 mM MgCl 2 , 0.1 M DTT, RNase OUT, Superscript IH RT) was added thereto, the resulting solution was subjected to reverse transcription at 50 ° C for 50 minutes and 85 ° C for 20 minutes.
- To the reaction solution was added 1 ⁇ Jt of RNAse H and kept at 37 ° C for 20 minutes to thereby synthesize cDNA.
- SEQ ID NO: 7 of pax3-l and SEQ ID NO: 8 of ⁇ ax3-2 for Pax3 amplification SEQ ID NO: 9 of snail- 1 and SEQ ID NO: 10 of snail-2 for Snail amplification
- the soft tissue filler composition for injection of the present invention exerts prolonged therapeutic effects without causing any immune response and side-effect due to the use of autologous dermal cells. Further, since it completely rules out the risk of the use of animal-derived serum through in vitro serum-free cultivation and immediately shows therapeutic effects by producing a large quantity of collagen, the soft tissue filler composition for injection of the present invention can be effectively used for improving the skin tone and resilience as well as smoothing and removing wrinkles and scars.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Dermatology (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Rheumatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
La présente invention concerne un procédé de préparation d'une composition de charge de tissus mous pour injection, comprenant les étapes suivantes: 1) la digestion d'un tissu dermique autologue isolé de la peau du patient et la séparation en cellules isolées; 2) la culture et la prolifération des cellules dermiques isolées en culture in vitro en milieu exempt de sérum pour obtenir un matériau autologue de culture cellulaire dérivé du derme contenant des cellules souches de fibroblaste dermique, des cellules amplificatrices transitoires de fibroblaste dermique, et des fibroblastes et du collagène dermique; 3) la centrifugation du matériau autologue de culture cellulaire dérivée du derme pour séparer une pastille cellulaire autologue dérivée du derme; et 4) la suspension de la pastille cellulaire autologue dérivée du derme dans une injection de glucose ou une injection aléatoire pour obtenir une suspension pour injection. L'invention concerne également une composition de charge de tissus mous pour injection préparée par le procédé selon l'invention.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2006-0057697 | 2006-06-26 | ||
| KR1020060057697A KR20070122316A (ko) | 2006-06-26 | 2006-06-26 | 주사용 인체 연조직 충전제 및 이의 제조 방법 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2008002064A1 true WO2008002064A1 (fr) | 2008-01-03 |
Family
ID=38845779
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/KR2007/003100 WO2008002064A1 (fr) | 2006-06-26 | 2007-06-26 | Composition de charge de tissus mous pour injection et son procédé de préparation |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR20070122316A (fr) |
| RU (1) | RU2396084C1 (fr) |
| WO (1) | WO2008002064A1 (fr) |
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| WO2011140323A1 (fr) * | 2010-05-07 | 2011-11-10 | Fibrocell Science, Inc. | Formulations d'unités de dosage de fibroblastes dermiques autologues |
| WO2012051505A1 (fr) * | 2010-10-14 | 2012-04-19 | Fibrocell Science, Inc. | Traitement de cordes vocales par une formulation de fibroblastes dermiques autologues |
| US9987473B2 (en) | 2009-12-18 | 2018-06-05 | Srgi Holdings, Llc | Skin treatment device and methods |
| US10076354B2 (en) | 2010-12-17 | 2018-09-18 | Srgi Holdings, Llc | Pixel array medical devices and methods |
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| CN110269868A (zh) * | 2019-06-12 | 2019-09-24 | 江苏艾尔康生物医药科技有限公司 | 一种含有自体真皮成纤维细胞透明质酸凝胶剂的构建方法 |
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| CN113181219A (zh) * | 2021-03-04 | 2021-07-30 | 纳美细胞科技有限公司 | 用于除皱的细胞制剂及其制备方法 |
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| CN114344454A (zh) * | 2022-01-21 | 2022-04-15 | 郑州市金水区蕾娜斯医疗美容门诊部 | 自体脂肪胶原蛋白注射剂的制备方法 |
| US11490952B2 (en) | 2015-08-31 | 2022-11-08 | Srgi Holdings, Llc | Pixel array medical devices and methods |
| US11564706B2 (en) | 2019-10-28 | 2023-01-31 | Srgi Holdings, Llc | Pixel array medical systems, devices and methods |
| US11673968B2 (en) | 2014-06-26 | 2023-06-13 | Hoffmann-La Roche Inc. | Anti-BRDU antibodies and methods of use |
| CN116474172A (zh) * | 2023-04-17 | 2023-07-25 | 广州市麦施缔医疗科技有限公司 | 表皮干细胞与脂肪混合注射方法和隆胸及丰臀再生方法 |
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Cited By (62)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US9987473B2 (en) | 2009-12-18 | 2018-06-05 | Srgi Holdings, Llc | Skin treatment device and methods |
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| KR20070122316A (ko) | 2007-12-31 |
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