US20100216206A1 - Method for Preparing an Acellular Organic Tissue for Revitalisation and Device for Implementing Said Method - Google Patents

Method for Preparing an Acellular Organic Tissue for Revitalisation and Device for Implementing Said Method Download PDF

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Publication number
US20100216206A1
US20100216206A1 US12/601,306 US60130608A US2010216206A1 US 20100216206 A1 US20100216206 A1 US 20100216206A1 US 60130608 A US60130608 A US 60130608A US 2010216206 A1 US2010216206 A1 US 2010216206A1
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Prior art keywords
tissue
needles
needle
holes
organic tissue
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US12/601,306
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English (en)
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Maurizio Marzaro
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TELEA BIOTECH Srl
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Maurizio Marzaro
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Publication of US20100216206A1 publication Critical patent/US20100216206A1/en
Assigned to TELEA BIOTECH S.R.L. reassignment TELEA BIOTECH S.R.L. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: MARZARO, MAURIZIO
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
    • 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
    • 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
    • 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
    • A61L27/38—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 containing added animal cells
    • 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
    • A61L27/56—Porous materials, e.g. foams or sponges
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin

Definitions

  • the invention relates to a method for preparing an acellular tissue for revitalisation and also concerns a device for implementing said method for preparing a tissue for revitalisation.
  • tissue engineering The creation of biological substitutes that are prepared in the laboratory and then transplanted into animal or human recipients is a medical procedure known by the name of “tissue engineering”.
  • tissues for grafting are prepared in the laboratory by implanting cells into a matrix consisting of an inorganic supporting medium generally called a “scaffold”.
  • the scaffold must naturally then undergo a process of degradation until it disappears completely and is replaced by the regenerated tissue, which is facilitated by the cells implanted in said scaffold.
  • Transplants can be obtained using this method with either artificial or natural scaffolds (i.e. from a “donor”) obtainable from humans or animals, such as the oesophageal wall.
  • a “donor” obtainable from humans or animals, such as the oesophageal wall.
  • the tissue To use a natural scaffold harvested from a donor for transplanting into another human being, the tissue must be treated first to eliminate all the cells existing between the fibres of the connective tissue, and then to reimplant human cells belonging to the intended recipient of the graft (the “host”) in order to avoid rejection phenomena.
  • a scaffold i.e. an acellular matrix, starting from tissues harvested from a donor
  • a fluid containing enzymatic substances capable of digesting and destroying the living cells contained within the tissue without damaging the tissue's connective fibres.
  • tissue, or scaffold After creating an acellular tissue matrix, ready to receive the cells obtained from the host, said tissue, or scaffold, is prepared in a so-called “Petri dish” (or similar container), which is a tray commonly used in biological laboratories, on the bottom of which the tissue to revitalise is rested.
  • the tissue is revitalised by implanting stem cells from the future recipient and nourishing them with a cell culture broth that feeds the cells, keeping them alive and enabling them to multiply and become disseminated.
  • the stem cells initially placed on the upper surface of the tissue move through the natural interstitia in the tissue of the scaffold—interstitia that were previously occupied by the donor's cells.
  • the living cells After a given period of time, under controlled temperatures and in the presence of the nutritional substances contained in the culture broth, the living cells reposition themselves in the interstitia of the tissue, which is then ready for transplantation into the host organ.
  • the cells generally used to revitalise the scaffold are stem cells, which subsequently become differentiated (or may have already done so) and acquire the specific function of the organ in which the revitalised tissue is grafted.
  • the success or failure of the transplantation of the tissue treated in this way depends on a capillary diffusion of the cells through the tissue matrix.
  • the object of the present invention is to develop a method for preparing acellular tissues that overcomes the above-mentioned drawbacks.
  • the object of the invention is to develop a method for preparing acellular organic tissue so that, when said tissue is revitalised with stem cells, it is easier for said cells to penetrate and colonise every possible space in the network of connective tissue fibres, so as to substantially recreate the same conditions of the tissue before it was devitalised.
  • Another object of the invention is to obtain a significant and important reduction in the treatment time needed to revitalise the acellular scaffold once the living cells have been added in order to prepare the tissue for transplantation.
  • the objects of the invention are achieved by a method for preparing an acellular organic tissue according to the contents of claim 1 .
  • this method consists in the creation of a plurality of holes in the surface of the tissue being prepared; these holes penetrate at least through a portion of the thickness, and preferably through the full thickness of the tissue concerned.
  • holes are obtained by means of a device containing needles with a suitable current passing therethrough and without inducing any alteration (tearing, necrosis, reduction or increase in thickness, changes in fluid content, or coagulation) in the connective tissue surrounding the hole being created.
  • the holes can be made through the thickness of the tissue being treated using various devices and methods, provided that the preparation of these holes does not cause any deterioration of the connective tissue surrounding the hole and of the scaffold in general.
  • the object of the invention and the best results in qualitative terms for the holes created in the tissue are achieved by applying a high-frequency voltage (generally 4 MHz) to the tip of each needle used to create each hole, so as to induce the passage of a weak electric current, but strong enough to break the bonds between the molecules in the connective tissue, thereby creating a hole, without inducing any breakage of the molecules.
  • a high-frequency voltage generally 4 MHz
  • needles of very narrow gauge e.g. in the order of 50 ⁇ M, but sufficient to facilitate the penetration of the cells inside said holes to revitalise the surrounding tissue.
  • creating numerous holes means preparing new routes for grafting cells into the deepest parts of the tissue, thus ensuring the complete revitalisation of the tissue concerned.
  • the thickness of the tissues that can be prepared for transplantation there is practically no limit to the thickness of the tissues that can be prepared for transplantation, since the holes can be made throughout the thickness of the tissue and over its entire surface, enabling its complete revitalisation because the living cells reimplanted in the acellular scaffold can penetrate throughout the tissue.
  • the invention also includes a device for implementing the method for preparing is the organic tissue according to the present invention, the main characteristics of which are expressed in claim 11 .
  • FIG. 1 shows a schematic cross-section of the device comprising a holder with an array of needles resting on the thickness of the tissue being prepared for revitalisation;
  • FIG. 2 shows one of the needles in the holder
  • FIG. 3 shows the layout of the array of needles
  • FIG. 4 shows a schematic view of the device for displacing the needle holder.
  • a previously-treated acellular organic tissue is deposited on the bottom of a Petri dish (or similar container) so that it lies spread out on a flat surface.
  • a plurality of needles such as the one indicated by the numeral 1 and shown in FIG. 2 , is arranged in an array, e.g. to form a square, indicated as a whole by the numeral 3 in FIG. 3 , so as to ensure an orderly arrangement of needles that are preferably separated by the same distance, i.e. they are equidistant from one another.
  • each needle 1 is connected electrically, e.g. by means of a metal conductor plate 2 attached to the holder 20 of said array of needles. Said plate 2 is connected to an electric wire 21 that in turn receives the output from a generator 4 .
  • Said generator 4 is a voltage generator, preferably generating 200-230 Volts, but at a wave frequency of 4 MHz, which is obtained by using electronic circuits, that are well known and consequently not described here for the sake of brevity.
  • the voltage sine wave available at the output 41 of the generator 4 is preferably a distorted sine wave and consequently with harmonics at least of the first, second and third order.
  • the power of the generator 4 is adjusted so that the current available at the tip is of each electrode 1 comes between 2 and 2.5 mA.
  • This energy is just enough to break the bonds between the molecules affected by the passage of the current, while in the surrounding area it causes no breakdown, tearing, necrosis, reduction or increase in thickness, change in fluid content fluid, coagulation or other tissue degeneration.
  • this opening created in the molecular bonds equates to the creation of a tiny hole that, in practical terms, is the same diameter as each needle 1 , i.e. around 50-55 ⁇ m.
  • needles of a different, larger or smaller gauge may be used, provided that the user bears in mind that the minimum gauge of the needle cannot be smaller than the diameter of the cells used for revitalisation.
  • the holder 20 of the array of needles 3 is then pushed in the direction in which the needles point and proceeds at a sufficiently slow pace such that, as the needle moves forward, the tip of the needle finds the hole already created by the flow of current and the consequent rupture of the molecular bonds.
  • the needles 1 penetrate preferably but not necessarily obliquely to the surface 51 of the scaffold 5 , in order to increase the length of the holes and consequently obtain the maximum channelling effect in the scaffold.
  • the perforation procedure must naturally be repeated all over the surface 51 of the scaffold in order to obtain a homogeneous distribution of the holes throughout the thickness and over the entire useful surface of the tissue for transplantation.
  • the invention includes a device for making the holes that is advantageously provided with means 30 for displacing the holder 20 along three Cartesian axes, i.e. along the vertical, or oblique axis Z, and along the Cartesian axes X and Y parallel to the plane of the surface 51 and shown schematically in FIG. 4 .
  • the holder 20 can be moved and the procedure can be repeated in an orderly manner so as to cover the entire surface 51 .
  • the procedure can be repeated automatically and sequentially, and with the utmost precision.
  • said acellular tissue can be placed in a Petri dish, or similar container, where the living cells can then be added, which are generally stem cells from the host individual intended to receive the graft. Suitably nourished with a culture broth, said stem cells can quickly and easily occupy all the holes made by the needles 1 , thereby ensuring a complete and effective revitalisation of the entire tissue for transplantation.
US12/601,306 2007-05-31 2008-04-16 Method for Preparing an Acellular Organic Tissue for Revitalisation and Device for Implementing Said Method Abandoned US20100216206A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
IT000159A ITVI20070159A1 (it) 2007-05-31 2007-05-31 Metodo di preparazione alla rivitalizzazione di un tessuto organico a-cellulare e dispositivo atto a realizzare tale metodo
ITVI2007A000159 2007-05-31
PCT/IB2008/000921 WO2008146106A2 (en) 2007-05-31 2008-04-16 Method and device for preparing an acellular organic tissue

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2008/000921 A-371-Of-International WO2008146106A2 (en) 2007-05-31 2008-04-16 Method and device for preparing an acellular organic tissue

Related Child Applications (1)

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US13/780,856 Division US10195307B2 (en) 2007-05-31 2013-02-28 Method for preparing an acellular organic tissue for revitalisation

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US20100216206A1 true US20100216206A1 (en) 2010-08-26

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US12/601,306 Abandoned US20100216206A1 (en) 2007-05-31 2008-04-16 Method for Preparing an Acellular Organic Tissue for Revitalisation and Device for Implementing Said Method
US13/780,856 Active US10195307B2 (en) 2007-05-31 2013-02-28 Method for preparing an acellular organic tissue for revitalisation

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US (2) US20100216206A1 (it)
EP (1) EP2164536B8 (it)
JP (1) JP5412688B2 (it)
KR (1) KR101555069B1 (it)
CN (1) CN101687064B (it)
AT (1) ATE552866T1 (it)
BR (1) BRPI0813396B8 (it)
CA (1) CA2688626C (it)
CU (1) CU23797A3 (it)
DK (1) DK2164536T3 (it)
ES (1) ES2385116T3 (it)
IN (1) IN266807B (it)
IT (1) ITVI20070159A1 (it)
PL (1) PL2164536T3 (it)
RU (1) RU2430745C1 (it)
WO (1) WO2008146106A2 (it)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USD856517S1 (en) 2016-06-03 2019-08-13 Musculoskeletal Transplant Foundation Asymmetric tissue graft
USD895812S1 (en) 2018-09-07 2020-09-08 Musculoskeletal Transplant Foundation Soft tissue repair graft
US10813743B2 (en) 2018-09-07 2020-10-27 Musculoskeletal Transplant Foundation Soft tissue repair grafts and processes for preparing and using same
US10945831B2 (en) 2016-06-03 2021-03-16 Musculoskeletal Transplant Foundation Asymmetric tissue graft

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT1398807B1 (it) * 2009-10-29 2013-03-18 Marzaro Metodo di preparazione di un tessuto organico a-cellulare di origine umana o animale per la rivitalizzazione
IT201900003299A1 (it) 2019-03-07 2020-09-07 Telea Biotech S R L Metodo di preparazione alla rivitalizzazione di un tessuto acellulare

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US1394171A (en) * 1921-05-02 1921-10-18 Mary E Hall Electric needle
US5112354A (en) * 1989-11-16 1992-05-12 Northwestern University Bone allograft material and method
US5658234A (en) * 1995-07-24 1997-08-19 J. D. Technologies, Inc. Method for treating tumors
US20010003155A1 (en) * 1997-01-22 2001-06-07 Rockley Paul W. Rapid pulse phaco power for burn free surgery
US6262532B1 (en) * 1998-03-31 2001-07-17 Samsung Display Devices Co., Ltd. Plasma display device with electrically floated auxiliary electrodes

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US6479064B1 (en) * 1999-12-29 2002-11-12 Children's Medical Center Corporation Culturing different cell populations on a decellularized natural biostructure for organ reconstruction
US6565532B1 (en) * 2000-07-12 2003-05-20 The Procter & Gamble Company Microneedle apparatus used for marking skin and for dispensing semi-permanent subcutaneous makeup
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Publication number Priority date Publication date Assignee Title
US1394171A (en) * 1921-05-02 1921-10-18 Mary E Hall Electric needle
US5112354A (en) * 1989-11-16 1992-05-12 Northwestern University Bone allograft material and method
US5658234A (en) * 1995-07-24 1997-08-19 J. D. Technologies, Inc. Method for treating tumors
US20010003155A1 (en) * 1997-01-22 2001-06-07 Rockley Paul W. Rapid pulse phaco power for burn free surgery
US6262532B1 (en) * 1998-03-31 2001-07-17 Samsung Display Devices Co., Ltd. Plasma display device with electrically floated auxiliary electrodes

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USD856517S1 (en) 2016-06-03 2019-08-13 Musculoskeletal Transplant Foundation Asymmetric tissue graft
US10945831B2 (en) 2016-06-03 2021-03-16 Musculoskeletal Transplant Foundation Asymmetric tissue graft
USD895812S1 (en) 2018-09-07 2020-09-08 Musculoskeletal Transplant Foundation Soft tissue repair graft
US10813743B2 (en) 2018-09-07 2020-10-27 Musculoskeletal Transplant Foundation Soft tissue repair grafts and processes for preparing and using same
US11642216B2 (en) 2018-09-07 2023-05-09 Musculoskeletal Transplant Foundation Soft tissue repair grafts and processes for preparing and using same

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Publication number Publication date
CU23797A3 (es) 2012-03-15
KR101555069B1 (ko) 2015-09-22
EP2164536B1 (en) 2012-04-11
CN101687064B (zh) 2014-06-04
ITVI20070159A1 (it) 2008-12-01
RU2430745C1 (ru) 2011-10-10
KR20100019524A (ko) 2010-02-18
ATE552866T1 (de) 2012-04-15
JP2010527724A (ja) 2010-08-19
WO2008146106A3 (en) 2009-05-07
IN266807B (it) 2015-06-03
BRPI0813396B1 (pt) 2018-07-03
WO2008146106A2 (en) 2008-12-04
DK2164536T3 (da) 2012-07-23
PL2164536T3 (pl) 2012-11-30
BRPI0813396A2 (pt) 2015-10-27
BRPI0813396B8 (pt) 2021-06-22
CA2688626A1 (en) 2008-12-04
CN101687064A (zh) 2010-03-31
EP2164536A2 (en) 2010-03-24
ES2385116T3 (es) 2012-07-18
JP5412688B2 (ja) 2014-02-12
RU2009149666A (ru) 2011-07-10
WO2008146106A9 (en) 2009-11-19
US10195307B2 (en) 2019-02-05
CA2688626C (en) 2013-01-22
EP2164536B8 (en) 2012-04-25
US20180339086A1 (en) 2018-11-29

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