WO2009044408A1 - Membrane amniotique traitée et sa méthode de traitement - Google Patents

Membrane amniotique traitée et sa méthode de traitement Download PDF

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Publication number
WO2009044408A1
WO2009044408A1 PCT/IN2008/000405 IN2008000405W WO2009044408A1 WO 2009044408 A1 WO2009044408 A1 WO 2009044408A1 IN 2008000405 W IN2008000405 W IN 2008000405W WO 2009044408 A1 WO2009044408 A1 WO 2009044408A1
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Prior art keywords
process according
skin
tissue
membrane
amniotic membrane
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PCT/IN2008/000405
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English (en)
Inventor
Soma Guhathakurta
Mammen Cherian Kotturathu
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International Centre For Cardio Thoracic And Vascular Diseases (A Unit Of Frontier Lifeline Pvt. Ltd)
Department Of Biotechnology (A Department Of Science And Technology, Government Of India)
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Application filed by International Centre For Cardio Thoracic And Vascular Diseases (A Unit Of Frontier Lifeline Pvt. Ltd), Department Of Biotechnology (A Department Of Science And Technology, Government Of India) filed Critical International Centre For Cardio Thoracic And Vascular Diseases (A Unit Of Frontier Lifeline Pvt. Ltd)
Publication of WO2009044408A1 publication Critical patent/WO2009044408A1/fr

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Classifications

    • 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
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like

Definitions

  • amniotic membrane ie both amnion and chorion
  • a fresh fetal membrane ie both amnion and chorion
  • amniotic membrane as a patch graft in the treatment of acute ocular burns.
  • Biological coverages should have the following characteristic.
  • Biological dressings have the following disadvantages also - They exhibit an early graft rejection and a rapid, acute bacterial colonization of the injured area.
  • Xenograft bovine, ovine, canine, porcine; cheap and readily available; used fresh, frozen or lyophilized.
  • Collagen products gel, sponges or other different plaques.
  • Allograft and xenografts possess numerous positive characteristics, but their use is sometimes limited, especially where tissue banks have not been developed.
  • Amniotic membrane has been used with variable success as a material for burn injury coverage.
  • the amnion is a unique membrane, composed of a single layer of cells, which completely line the cavity in which the foetus grows and develops. It is formed from the same small group of cells, which are formed when the ovum fertilised by the sperm begins to divide. These cells have the potential to develop into all the cells of the human body and under the action of growth factors, and by a process which is as yet undefined, the cells will differentiate into either muscles, bones, heart, liver or whatever structure is required. We now have reason to believe that the cells which go to form the amnion retain this pluripotential characteristic of being able to differentiate, at least in part, into other tissues.
  • Amniotic membrane can be used for superficial bums, deep burns, after necrectomy, (excision of the necrotic tissue) on extensive granulating wound surfaces, on autografts, in donor regions, and after dermoabrasion.
  • Amniotic ⁇ membrane is readily available and does not present immunological problems. It does not cause allergic responses and reduces water loss. The risk of the transmission of some viral infections is there. Bacterial examinations performed with burn wounds covered with amniotic membrane showed low or no bacterial colonization of the burn surface. It is concluded that amniotic membrane should be more widely used in this particular aspect of burn treatment.
  • FMs are also known as amniotic membrane.2
  • Anatomically, FMs consist of two loosely connected layers, an inner one of amnion and an outer one of chorion.
  • the inner amnion layer is composed of cuboidal, flattened epithelial cells and mesenchymal connective tissue.
  • the outer chorion is composed of fairly thick transitional epithelium.
  • Amnion is thin and shiny in contrast to chorion, which is less homogeneous and dull.3 Foetal membrane can be used into to (amnion + chorion) or only as amnion (epithelium + base membrane).4 Amniotic membrane is also comparable with honey, for its good healing effect.
  • amniotic membrane Another unique feature of the amniotic membrane is the complete lack of expression of surface antigens responsible for mounting an immune reaction. Thus, the amnion does not induce an immune response when transplanted into a "foreign" site, a feature which is of major importance to the foetus. Some groups found immunogenicity may be in lower form in homologous amniotic membrane.
  • amniotic membrane exhibits histological structure similar to that of skin. -low much ever, be the advantages, simultaneously there exists disadvantage of the use of amniotic membrane is that there is some risk of viral infection transmission, e.g. hepatitis, syphilis and AIDS.
  • amniotic membrane There are two varieties of amniotic membrane are mainly used in the clinical procedures.
  • amnion alone epidermal + basic membrane
  • Amniotic membrane have been use by surgeons, during preparation of deep burns for necrectomy(excision of necrotic tissue), to keep necrotic areas dry or after necrectomies leaving raw areas of flesh - frequently in combination with other temporary biological skin coverages and/or autografts.
  • Human amniotic membrane is derived from the fetal membranes which consist of the inner amniotic membrane made of single layer of amnion cells fixed to collagen-rich mesenchyme 6 to 8 cells thick loosely attached to chorion. It is composed of three layers: a single epithelial layer, thick basement membrane, and avascular stroma. Human amniotic membrane has been shown to contain collagen types III and V. It also contains collagen types IV and VIl similar to corneal epithelial basement membrane as well as fibronectin and laminin. Additionally, it contains fibroblast and other growth factors.
  • amniotic membrane is believed to be nonimmunogenic. Antibodies or cell-mediated immune response to amniotic membrane have not been demonstrated by few groups, suggesting low antigenicity. Therefore, the use ofsystemic immunosuppressives in AMT is not required.
  • Foetal membranes have been put to a variety of surgical membranes for skin coverage in the past. Amnion has been used service early nineteenth century for treating burns and skin ulcers and reported relief of pain, increased rate of epithelialisation and no infection. There have been several use of amnion as a temporary dressing for ulcers, burns and other denuded areas. Stored and desiccated membranes have not been found to offer any significant improvements over other more conventional sterile dressings. But when applied fresh or following preservation at 4°centigrade in isotonic saline, the amnion appears to have advantages which have been reported to include reduction in bacterial counts, relief of pain and hastened healing. The amnion has been postulated as producing factors which promote granulation tissue formation, neovascularisation and re- epithelialisation although this is a controversial area.
  • amniotic membrane can be understood from the basic properties of human foetal membranes.
  • FMs There is an absence of immunological rejection and the healing effects of FMs may be due to: a. Antibacterial factors b. Biological factors c. The biomechanical characteristics of FMs
  • amniotic cells were known to be pluripotential in their natural state i.e. during formation of the foetus. It was also known that human allografts of amniotic cells generated no significant immunological problems though substantially it is substituted.
  • the mechanism responsible for the rapid healing observed is due to the inhibition of the proteinase activity, thus reducing the inflammatory responses by reducing the infiltration of polymorphonuclear leukocytes.
  • Human amniotic epithelial cells do not express on their surfaces HLA-A, B, C, and DR antigens, or beta 2- microglobulin, which could further contribute to the lower inflammatory responses and relatively delayed rejection of this type of biological dressing.
  • the amniotic membrane-treatment method means the patient will be similarly treated as in above described conventional method by cleansing of the bum wounds with diluted povidone iodine in normal saline and covered with a layer of the amniotic membrane with the amnion side down over the whole of the affected areas. The wounds were similarly inspected.
  • Human amniotic membrane is one of the most effective biological dressings that is used in burn treatment. Using human amniotic membrane incorporating silver nitrate gives a better therapeutic effect than plain amniotic membranes. Silver nitrate incorporated into the membranes increases their maneuverability, provides easier application to the burned area and creates a bactericidal effect, therefore redudnq the risk of contamination and infection. But re-epithelialization is hindered.
  • One of the main advantages of wound coverage with only amniotic membrane is that it does not appear to discourage re-epithelization, reduces fluid, protein, heat and energy loss, increases mobility and most important this may be the ideal wound cover next to the patient's own skin.
  • silver nitrate- incorporated amniotic membrane
  • amniotic membrane since it is readily available and freely obtainable, has low preparation and storage costs that make it an ideal dressing to use, especially in countries where economic factors prevent the purchase of other types of dressings.
  • the silver nitrate is found to be toxic in nature and thereby is not enhance the growth eventhough it may be useful when it is used as part of dressing only. It is not useful when amniotic membrane is used for implantation.
  • the main obstacle encountered in organ transplantation is the immune rejection of the transplant.
  • the rejection phenomenon originates from antigenic differences between the cells of the recipient and the transplant, as well as from the natural immune response of the organism towards "non-self antigens.
  • Attempts to prolong the survival of allografts and xenografts, both in experimental models and in medical practice, have been mainly aimed at the supression of the immune apparatus of the recipient. This has been achieved by means of cytotoxic drugs, antimetabolites, corticosteroids and antilymphocytic serum.
  • the generalized immunosuppression is accompanied by undesirable toxic effects, decreased resistance to infection and reduction in the level of the haemopoietic stem cells.
  • Another possibility is to attenuate, or to abolish completely, the antigenicity of the graft with preservation of its biological functions.
  • the advantage of this approach is that the immune capacity of the recipient is not affected. Investigations along this line, have been, for the most part, unsuccessful. Most studies were performed with animals and have been subjected to clinical evaluation. Using laboratory animals, the treatment of allografts or xenografts in vitro, with cortisone, thalidomide or urethane prolonged their retention time by a factor of about two. The amount of drug locally applied to the skin was smaller than the amount required to achieve a similar effect by injecting the drug systemically.
  • the donor skin has been treated in vitro with streptokinase/streptodornase, or with RNA and DNA preparations of the recipient. Allograft survival was not prolonged by exposure of donor skin to transplantation antigens of the recipient. Minimal immune reaction was observed towards grafts in which cellular viability was destroyed in vitro treatment with formalin or cyanide or by freeze-drying. The majority of the dead grafts were retained by the host for a limited period of time. Amniotic membrane has been used to replace damaged tissue or as a dressing on top of existing tissue. It is attempted to use Grafts made of amniotic membrane — one of the protective layers surrounding the fetus — can be used to stimulate the regrowth of damaged skin.
  • amniotic membrane transplantation AMT
  • ophthalmology in the treatment of conjunctival epithelial defects after symblepharon (scarring and adhesions between palpebral and bulbar conjunctiva).
  • symblepharon scarring and adhesions between palpebral and bulbar conjunctiva.
  • caustic burns of the conjunctiva with corneal involvement were also treated successfully using amniotic membrane.
  • amniotic membrane delivers its beneficial effects on the ocular surface is not fully understood or known.
  • Amniotic membrane modulates levels of cytokines and growth factors and has also been shown to have unique properties, including pain reducing, fibrosis suppressing, antibacterial, and wound protecting. It is unclear whether amniotic membrane promotes limbal stem cell proliferation.
  • Amniotic membrane transplant was also successful in promoting the growth of corneal tissue in a patient whose scarring had caused the eyelid to adhere to the eye.
  • the 'antiadhesive' compounds in amniotic membrane tissue appear to help prevent this type of scarring.
  • the invention in general relates to the use of amniotic membrane as dressing, as graft and for transplantation.
  • the Uniqueness of the invention lies in the decellularisation and cross linking of the membrane prior to the use, which makes the membrane adaptable for adhering to the wound area and thereafter enhancing the growth of the neighbouring tissue which further allows the foreign membrane placed on the wound area to merge with the neighbouring skin.
  • the method begins with harvesting the membrane from caesarean section and mother is screened for infectious diseases before going for surgery.
  • a membrane piece is sent for RT- PCR studies for retrovirus identification. It is collected in balance salt solution with antifungal medication to prevent fungal contamination.
  • tissue 50gms of tissue is treated with 100ml of the solution of 1 % Deoxy cholic acid for 20- 50 hours alongwith ribonuclei enzyme treatment.
  • Acellular amniotic membrane for clinical use in eyes as corneal replacement and limbal cell growth have been done with trypsinization, and being practised by different groups .
  • Our preparation is different and we have found out that it is helping auto- logous cell and tissue growth wherever applied.
  • Cell adhesion molecules like laminin cadherin are active by this processing. Application of this will transform a full thickness skin burn of large area into re replacement of patients own skin with any scarring . It will encourage the adjacent skin stem cell to migrate in it and form new skin, thereby avoiding skin grafting in future as well as deformity of scar formation.
  • Amniotic membrane has thus inherent properties that are helpful in wound healing. Among other benefits it is thought to inhibit blood vessei growth in adjacent tissues, decrease cell death, reduce inflammation, and reduce scar tissue, which is vital for use in skin transplantation.
  • amniotic membrane graft "dissolved” away within few months, leaving healed corneal tissue in its stead whereas in the invention, we are attempting to regenerate a damaged skin with an application of treated amniotic membrane in the process described in the invention.
  • the bacteria are trapped in the thin fibrin matrix linking the collagen fibres of the graft with the collagen of the wound )ed.
  • the fibrin matrix provides an ideal substratum for migration of phagocytes and ensures that all the bacteria are within reach of the phagocytes.
  • amniotic membrane transplantation can reduce inflammation, promote healing, and decrease irritation in surface problems.
  • amniotic membrane consists of a single layer of cuboidal epithelial cells, a thick basement membrane and an avascular stromal matrix, loosely attached to the chorion. Amniotic membrane so processed has been found to exhibit the following features:
  • cytokines such as IL-4, 6 and 10
  • EGF EGF, FGF, TGF, HGF, and 2-macrobulin
  • IL-4, 6 and 10 EGF, FGF, TGF, HGF, and 2-macrobulin
  • Amniotic membrane can be used in a number of indications, either as a 'substrate' to replace the damaged ocular tissue or as a 'patch' (biological dressing), or a combination of both. Even it can be used as a covering for the heart when pericardium is lost due to surgery. By this covering, the second surgery of the heart becomes easier because of its inflammatory function. It is also very much useful for healing non-healing access diabetic foot ulcer and gangrenous foot ulcer.
  • the human body has considerable capacity for regeneration.
  • the skin with high rates of cell turnover are regenerated continually through out life.
  • the present invention provides methods and compositions to actively control the whole process of skin regeneration.
  • cells the smallest unit of life, are stimulated, propagate, differentiate, integrate with each other to physiologically repair the damaged skin or to regenerate the skin destroyed in various courses, such as trauma and diseases. These nascent skin then conjoin together to form a fully functional skin.
  • the general guidance for this intervention revealed in the present invention is that 1) preserving the injured or damaged skin, the viable cells in the remaining tissues should be preserved to a maximum extent; 2) removing necrotic cells or skin as early as possible; 3) activating and propagating the regenerative cells in an environment mimicking the their own native physiological conditions; and 4) administering regulators for cell growth and differentiation to the regenerating organ to direct proper, physiological repair of tissues.
  • Neutrophiles then macrophages, migrate into the wound, characterizing acute inflammation. These inflammatory cells provide phagocytosis of bacteria and debridement of injured tissue. This is proceeded by chronic inflammation where lymphocytes and monocytes infiltrate the wound site. The latter become macrophages, which are considered the main coordinators of adult wound healing.
  • Neutrophils phagocytize contaminating bacteria and digest the fibrin matrix in preparation for new tissue. They also secrete vasodilatory mediators and cytokines •that activate fibroblasts and keratinocytes and attract macrophages to the injury site.
  • Macrophages phagocytize potential pathogens, debride the wound, and secrete cytokines and growth factors such as fibroblast growth factors (FGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF-a), interleukin-1 (IL-1) and interferon-gamma (IFN-. gamma.).
  • FGF fibroblast growth factors
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • TNF-a tumor necrosis factor
  • IL-1 interleukin-1
  • IFN-. gamma interferon-gamma
  • Amnion becomes incorporated into, and becomes part of, the host
  • reference is made to the use of human amnion although it is to be understood that the invention is not restricted to the use only of human amnion.
  • the human amnion is easily obtained in the fresh state from caesarean sections in the maternity wards, and can be kept in organ culture. We have further found that the amnion induces healing of burnt skin and in time is indistinguishable from the original skin. Accordingly, in one aspect the present invention provides the use of human or animal amniotic cells to regenerate or replace diseased or damaged skin.
  • the healing ECM also contains many glycoproteins, including fibronectin, and tenascin. Fibronectin promotes substrate adhesion, whereas tenascin facilitates substrate migration by antagonizing fibronectin.
  • fibroblasts and endothelial cells convert dissolved molecular oxygen to superoxide, which is important in resistance to wound infection as well as oxidative signaling in further stimulation of growth factor production.
  • epithelization occurs in order to re- epithelialize the wound edges.
  • an epidermal covering composed predominantly keratinocytes begins to migrate and undergo stratification and differentiation to reconstitute the barrier function of epidermis.
  • This process also promotes extracellular matrix (ECM) production, growth factor and cytokine expression and angiogenesis through the release of growth factors such as keratinocyte growth factor (KGF).
  • ECM extracellular matrix
  • KGF keratinocyte growth factor
  • Keratinocytes stimulate angiogenesis by releasing basic fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF). They also secrete TGF-.alpha.
  • FGF basic fibroblast growth factor
  • VEGF vascular endothelial growth factor
  • fibroblasts which acts as a chemoattractant and mitogen
  • PDGF PDGF which is involved in matrix production.
  • Further migration and proliferation of fibroblasts lead to the replacement of proteoglycan in the ECM with collagen deposition.
  • endothelial cell proliferation creates neovascularization, i.e., angiogenesis.
  • amniotic membrane processed by the present invention if placed on a skin wound, acts as a bioactive scaffold which attracts adjacent normal skin's stem cells from the basement membrane to migrate.

Abstract

L'invention porte sur une méthode de traitement d'une membrane amniotique et sur l'utilisation d'une membrane amniotique traitée, en tant que pansement, greffe ou transplant, et sur une méthode et des compositions contrôlant activement la totalité d'un processus de régénération de la peau. La singularité de l'invention réside dans la décéllularisation avant utilisation et la réticulation de la membrane, ce qui facilite son adhérence à la surface de la plaie, favorise la croissance des tissus voisins et permet de plus à une membrane étrangère placée sur la plaie de fusionner avec la peau du voisinage.
PCT/IN2008/000405 2007-10-03 2008-06-25 Membrane amniotique traitée et sa méthode de traitement WO2009044408A1 (fr)

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US20130209524A1 (en) * 2012-02-14 2013-08-15 AFcell Medical Method of using amnion allograft in heart transplant surgery
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CN103623455A (zh) * 2013-11-29 2014-03-12 东方康瑞(北京)科技发展有限责任公司 一种生物护创膜的制备方法
CN103861151A (zh) * 2014-03-27 2014-06-18 成都青山利康药业有限公司 一种脱细胞胎盘基质材料的制备方法
WO2014113733A1 (fr) 2013-01-18 2014-07-24 Mimedx Group, Inc. Procédés de traitement d'états cardiaques
US8840665B2 (en) 2010-06-11 2014-09-23 Liventa Bioscience, Inc. Method of tendon repair with amnion and chorion constructs
US8961617B2 (en) 2012-03-08 2015-02-24 Liventa Bioscience, Inc. Amnion and chorion constructs and uses thereof in abdominal surgery
US9433490B2 (en) 2010-12-22 2016-09-06 University Of Florida Research Foundation, Inc. Multilayered implant materials derived from amniotic membrane, methods of making the multilayered implant materials, and method of using multilayered implant materials
CN105963784A (zh) * 2016-05-05 2016-09-28 重庆大学 一种恢复脱细胞血管残余应力和残余应变的方法及其试剂
US9498327B1 (en) 2013-03-05 2016-11-22 Biodlogics Llc Repair of tympanic membrane using human birth tissue material
US9585983B1 (en) 2011-10-12 2017-03-07 BioDlogics, LLC Wound covering and method of preparation
US9770472B1 (en) 2013-03-08 2017-09-26 Brahm Holdings, Llc Organ jacket and methods of use
US9789138B1 (en) 2013-03-06 2017-10-17 BioDlogics, LLC Neural repair construct and method of use
US9795638B1 (en) 2013-03-16 2017-10-24 BioDlogics, LLC Cardiothoracic construct and methods of use
CN107296041A (zh) * 2017-07-02 2017-10-27 江西瑞济生物工程技术股份有限公司 一种新鲜羊膜保存液及新鲜羊膜保存方法与应用
US9855301B1 (en) 2013-03-13 2018-01-02 Biodlogics Llc Human birth tissue laminate and methods of use
US10265438B1 (en) 2014-11-03 2019-04-23 BioDlogics, LLC Methods and compositions for the repair and replacement of connective tissue
CN110404119A (zh) * 2019-08-29 2019-11-05 陈万娟 羊膜组织工程去免疫原皮肤支架的制备方法
US10765705B2 (en) 2014-11-24 2020-09-08 Prime Merger Sub, Llc Visco-supplement compositions, and methods of use thereof
US10905800B1 (en) 2013-01-29 2021-02-02 BioDlogics, LLC Ocular covering and method of use
US20210369784A1 (en) * 2010-02-18 2021-12-02 Osiris Therapeutics, Inc. Immunocompatible amniotic membrane products

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US20210369784A1 (en) * 2010-02-18 2021-12-02 Osiris Therapeutics, Inc. Immunocompatible amniotic membrane products
US8840665B2 (en) 2010-06-11 2014-09-23 Liventa Bioscience, Inc. Method of tendon repair with amnion and chorion constructs
US9433490B2 (en) 2010-12-22 2016-09-06 University Of Florida Research Foundation, Inc. Multilayered implant materials derived from amniotic membrane, methods of making the multilayered implant materials, and method of using multilayered implant materials
US20130144386A1 (en) * 2011-09-30 2013-06-06 NuTech Spine, Inc Osteoconductive Implants and Methods of Using Same
US9539104B2 (en) * 2011-09-30 2017-01-10 NuTech Spine, Inc. Osteoconductive implants and methods of using same
US9486316B2 (en) * 2011-09-30 2016-11-08 NuTech Spine, Inc. Osteoconductive implants and methods of using same
US20130138222A1 (en) * 2011-09-30 2013-05-30 NuTech Spine, Inc. Osteoconductive Implants and Methods of Using Same
US9585983B1 (en) 2011-10-12 2017-03-07 BioDlogics, LLC Wound covering and method of preparation
US20130209524A1 (en) * 2012-02-14 2013-08-15 AFcell Medical Method of using amnion allograft in heart transplant surgery
US8961617B2 (en) 2012-03-08 2015-02-24 Liventa Bioscience, Inc. Amnion and chorion constructs and uses thereof in abdominal surgery
US20150238540A1 (en) * 2013-01-18 2015-08-27 Thomas J. Koob Methods for treating cardiac conditions
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