WO2022079702A1 - Composition exempte de formol pour conserver et rendre viables des tissus - Google Patents

Composition exempte de formol pour conserver et rendre viables des tissus Download PDF

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Publication number
WO2022079702A1
WO2022079702A1 PCT/IB2021/060372 IB2021060372W WO2022079702A1 WO 2022079702 A1 WO2022079702 A1 WO 2022079702A1 IB 2021060372 W IB2021060372 W IB 2021060372W WO 2022079702 A1 WO2022079702 A1 WO 2022079702A1
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WO
WIPO (PCT)
Prior art keywords
agent
free composition
formalin
specimen
formaldehyde
Prior art date
Application number
PCT/IB2021/060372
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English (en)
Spanish (es)
Inventor
Roberto Javier RUEDA ESTEBAN
Original Assignee
Universidad De Los Andes
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidad De Los Andes filed Critical Universidad De Los Andes
Publication of WO2022079702A1 publication Critical patent/WO2022079702A1/fr
Priority to CONC2022/0007327A priority Critical patent/CO2022007327A2/es

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • 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
    • A61L9/00—Disinfection, sterilisation or deodorisation of air

Definitions

  • the composition of the present development belongs to the field of tissue preservation, particularly to compositions to preserve and make viable tissues in the disciplines of mortuary science, anatomy, surgery, pathology, among others.
  • the present development provides a formalin-free composition that improves the mechanical characteristics of tissues and reduces sensory discomfort for users, thus making viable specimens for surgical education and simulation.
  • compositions that have been developed as alternatives to formaldehyde for tissue preservation, we can mention patent US5679333, which discloses a formaldehyde-free composition for embalming bodies and histological preservation that contains an aqueous solution of ethanol, glyoxal (ethanedial) polyethylene glycol and dimethylsulfoxide (DMSO).
  • DMSO dimethylsulfoxide
  • the composition may include a humectant corresponding to 1,2-ethanediol, an antimicrobial corresponding to 1-hexadecylpyrimidinium chloride, and a chelating agent corresponding to ethylenediaminetetraacetic acid (EDTA).
  • EDTA ethylenediaminetetraacetic acid
  • patent US9393590 discloses a composition for tissue preservation that does not contain formaldehyde or glutaraldehyde, and that does not contain alcoholic solvents or compounds such as acetic acid that pose hazardous waste disposal problems and are potentially irritating to exposed people .
  • the preservative composition is aqueous and contains glyoxal (ethanedial) and dimethylsulfoxide (DMSO).
  • the composition may include a polymer corresponding to polyethylene glycol, a humectant corresponding to ethylene glycol, propylene glycol glycerol, or combinations thereof, an antimicrobial corresponding to 1-hexadecylpyrimidinium chloride, and a chelating agent corresponding to ethylenediaminetetraacetic acid (EDTA). ).
  • compositions and other similar ones reduce the concentration of formaldehyde or completely eliminate this component, they present technical challenges since they alter the characteristics of the fabric such as color, volume, flexibility and other aspects that are important for the adequate formation of the fibers. doctors and research in the disciplines of mortuary science, anatomy, surgery, pathology, among others. Additionally, some of the compositions offer short useful times of the preserved tissues, due for example to the formation of salt crystals in the specimens, or the use of the composition becomes limited due to the high costs of some or all of its components. .
  • compositions free of carcinogenic components such as formaldehyde
  • for the conservation and viability of fabrics that maintain the characteristics of the fabric such as color, volume and flexibility and that preserve the fabrics for long periods of time, facilitating medical training in anatomy and surgical simulation training.
  • the present development refers to a formalin-free composition to preserve and make viable tissues that comprises a moisturizing agent between 20 to 30% v/v; a preservative agent between 15 to 25% v/v; a sporicidal and fungicidal agent between 8 to 14% v/v; a bacteriostatic agent between 4 to 10% w/v; an insect repellent agent between 0.1 to 5% v/v in an aqueous medium.
  • the present development refers to a formalin-free composition to preserve and make viable tissues that comprises glycerin between 24 to 26% v/v; ethyl alcohol between 19 to 21% v/v; N-alkyldimethylbenzyl ammonium chloride between 9 to 1 l% v/v; carbamine between 3.5 to 5.5% w/v; citronella essential oil between 0.1 to 1.5% v/v and water.
  • the present development refers to a method for the conservation and viability of specimens that comprises putting the specimen in contact with the formalin-free composition that comprises a moisturizing agent between 20 to 30% v/v; a preservative agent between 15 to 25% v/v; a sponcidal and fungicidal agent between 8 to 14% v/v; a bacteriostatic agent between 4 to 10% w/v; an insect repellent agent between 0.1 to 5% v/v and an aqueous medium.
  • the formalin-free composition that comprises a moisturizing agent between 20 to 30% v/v; a preservative agent between 15 to 25% v/v; a sponcidal and fungicidal agent between 8 to 14% v/v; a bacteriostatic agent between 4 to 10% w/v; an insect repellent agent between 0.1 to 5% v/v and an aqueous medium.
  • the present development refers to a method for the preservation and viability of specimens that comprises immersing the specimen in the formalin-free composition for between 4 to 15 days.
  • the present development refers to a method for the conservation and viability of specimens that comprises injecting through any conduit of the specimen between 20 to 30mL of formalin-free composition for each IKg of weight of the specimen.
  • the present development refers to a method for the conservation and viability of specimens where the viability specimen is kept preserved by contacting the specimen with the formalin-free composition every 2 to 5 months for between 4 to 5 days. .
  • the present development refers to a process for obtaining a formaldehyde-free composition
  • a process for obtaining a formaldehyde-free composition comprising the steps: a) diluting a bacteriostatic agent in an aqueous medium until a homogeneous mixture is obtained; b) add to the homogeneous mixture a moisturizing agent, and a sporicidal and fungicidal agent; c) incorporating a preservative agent into the mixture; and d) add an insect repellent agent and stir until a homogeneous composition is obtained, where the steps are carried out at temperatures between 10°C to 30°C.
  • the present development refers to the use of a formalin-free composition for the conservation and viability of cadaveric specimens for education in human anatomy, veterinary or related areas, in medical training, in basic medical science anatomy with free specimens of carcinogenic agents such as formaldehyde and for the training of surgical skills through the medical-surgical simulation in areas of minimally invasive medical-surgical training.
  • FIG. 1 Flow diagram of the process steps to obtain the formalin-free composition.
  • FIG. Flow chart of the tissue viability method.
  • FIG. 3 Evaluation of the physical-mechanical characteristics of a fabric with the evaluated solutions.
  • A Specimen preserved in formalin-free composition
  • B Specimen previously fixed in a 4% formaldehyde solution and then preserved with the formalin-free composition
  • C Specimen fixed in a 4% formaldehyde solution.
  • FIG. 4 Evaluation of the unpleasant stimuli of users with the evaluated compositions.
  • A Specimen preserved in formalin-free composition
  • B Specimen previously fixed in a 4% formaldehyde solution and then preserved with the formalin-free composition
  • C Specimen fixed in a 4% formaldehyde solution.
  • the development involves a composition free of formalin or formaldehyde for the conservation and viability of fabrics that recovers mechanical and colorimetric characteristics of the fabric, reduces sensory discomfort for users when in contact with the fabrics, increases the useful life of the fabrics and prevents the use of carcinogenic reagents.
  • tissue preservation refers to the ability of the composition to preserve cadaveric tissues, avoiding colonization by pathogenic agents, preserving to a certain extent the mechanical characteristics of living tissue
  • tissue viability refers to to the ability to perform simulations for minimally invasive surgery procedures such as laparoscopy, thoracoscopy or arthroscopy.
  • the formalin-free composition comprises a moisturizing agent; a preservative agent; a sporicidal and fungicidal agent; a bacteriostatic agent and an insect repellent agent in an aqueous medium.
  • Moisturizing agent refers to compounds capable of attracting moisture to cells to complete the balance of cellular osmotic pressure and promote flexibility of the attached tissue.
  • the moisturizing agent favors the flexibility of the preserved tissues, reducing to a certain extent the rigidity generated in previously fixed specimens.
  • the moisturizing agent in the present development is selected from, but not limited to: glycerin, propylene glycol, and mixtures thereof. In one embodiment, the moisturizing agent is glycerin.
  • the moisturizing agent is found in the composition free of formaldehyde in a concentration between 20% and 30% v/v; between 20% to 22%v/v; between 22% to 24%v/v; between 24% to 26%v/v; between 26 to 28% v/v or between 28% to 30% v/v.
  • Preservative agent refers to compounds that increase stability and delay or prevent bacterial growth.
  • the preservative agent guarantees the stability of the preserved tissues, performs a bactericidal and preservative action, and has the ability to inhibit autolysis by paralyzing the enzymatic activity of the tissue and the bacterial action responsible for putrefaction.
  • the preserving agent in the present development is selected from, but not limited to, methanol, ethanol, isopropyl alcohol, and mixtures thereof.
  • the preservative agent is ethyl alcohol.
  • the preservative agent is found in the composition free of formaldehyde in a concentration between 15% and 25% v/v; between 15% to 19%v/v; between 19% to 21%v/v; between 21% to 23%v/v or between 23 to 25%v/v.
  • the sporicidal and fungicidal agent refers to compounds that simultaneously have activity to prevent the proliferation of bacteria and sporulated fungi and to prevent the growth of fungi or mold in tissues.
  • the sporicidal and fungicidal agent guarantees an environment free of molds and other fungal agents that can compromise or alter the stability of the fabrics.
  • the sporicidal and fungicidal agent in the present development is selected from, but not limited to, quaternary ammonium compounds (QACs) such as benzalkonium chloride, didecyldimethylammonium chloride, N-alkyldimethylbenzyl ammonium chloride, glutaraldehyde, and mixtures thereof.
  • QACs quaternary ammonium compounds
  • the sporicidal and fungicidal agent is added to the composition as an aqueous dilution between 30% to 50%, between 30% to 40% or between 40% to 50%.
  • the sporicidal and fungicidal agent is a 50% solution of N-alkyldimethylbenzyl ammonium chloride.
  • the sporicidal and fungicidal agent is found in the composition free of formaldehyde in a concentration between 8% and 14% v/v; between 8% to 9%v/v; between 9% to 1 l%v/v; between 11% to 12%v/v or between 12 to 14%v/v.
  • the bacteriostatic agent refers to compounds that, although they do not kill a bacterium, prevent their reproduction and population growth; bacteria age and die without leaving offspring.
  • Bactenostatic provides an aseptic environment ensuring tissue maintenance and a safe environment for the handler.
  • the bacteriostatic agent in the present development is selected from, but not limited to, urea, carbamine, carbonyldiamide, carbamic acid, aliphatic amine, sulfanilamide, and mixtures thereof.
  • the bacteriostatic agent is carbamine.
  • the bacteriostatic agent is found in the composition free of formaldehyde in a concentration between 3% and 10% w/v; between 3.5% to 5.5% w/v; between 5% to 8%p/v or between 8 to 10%p/v.
  • Insect repellent agent refers to substances or mixtures of substances of natural origin that prevent, eliminate, repel or mitigate insects and arthropods from climbing or climbing on any surface.
  • the insect repellent agent not only limits the possible invasion or infestation by insects of different types, associated with environments in which tissues are preserved, in some cases with a certain degree of decomposition, but also fulfills a virucidal function.
  • the insect repellent agent in the present development is an essential oil or mixture of essential oils that are selected from, but not limited to, citronella, eucalyptus, mint, lemon, basil, clove, thyme, grapefruit, oregano, lemongrass, geranium, lavender, cinnamon, bergamot, rosemary, cedar, lavender, pine and mixtures thereof.
  • the insect repellant agent is citronella essential oil.
  • the insect repellent agent is found in the formalin-free composition in a concentration between 0.1% and 5% v/v; between 0.1% to 1.5% v/v; between 1.5% to 3%v/v or between 3 to 5%v/v.
  • Another object of the development is to provide a process for obtaining the formalin-free composition described above.
  • the process to obtain the formalin-free composition comprises the steps of diluting a bacteriostatic agent in an aqueous medium until a homogeneous mixture is obtained; add to the homogeneous mixture a moisturizing agent, and a sporicidal and fungicidal agent; incorporate a preservative agent into the mixture; and finally add an insect repellent agent and stir until a formalin-free composition is obtained (FIG. 1).
  • the bactenostatic agent is a compound in solid state, which initially dissolves in the aqueous medium.
  • the initial addition of the bacteriostatic agent to the composition is carried out in the absence of other components to guarantee its complete dissolution in the composition, since if it is carried out in the presence of other components, its dissolution does not occur completely or may take longer.
  • the bacteriostatic agent, the moisturizing agent, and a sporicidal and fungicidal agent are added to the solution under the necessary stirring conditions to guarantee the complete homogenization of the components.
  • the preservative agent is added to the mixture, which produces an exothermic reaction that increases the temperature of the mixture and favors the dissolution of the previously incorporated components.
  • the repellent agent is added and it is stirred under the necessary conditions to guarantee the complete homogenization of the components to obtain the composition free of formaldehyde.
  • the process for obtaining the formalin-free composition is carried out at room temperature, which for the purposes of this development refers to temperatures between 10°C and 30°C.
  • a viability method refers to a method by which a previously fixed specimen can be recovered to the point of allowing simulation of minimally invasive surgical procedures in body cavities, joints, etc.
  • a specimen refers to to any tissue, organ or whole human body or any inert mammal, without difference in age at the time of death or mechanism thereof.
  • the specimen is previously fixed.
  • a fixed specimen refers to one that has been put in contact with fixing agents such as alcohols and/or formaldehyde or mixtures thereof.
  • the prior fixation of the specimen prevents the degradation of the tissues and allows its use in education for long periods of time at the expense of the loss of the mechanical properties of the living tissue and some associated risks, such as formaldehyde, as it is a known carcinogenic agent. .
  • the specimen is contacted by immersion in the formalin-free composition for 4 to 15 days, 4 to 8 days, or 8 to 15 days.
  • the specimen is contacted by injecting through any conduit of the specimen between 20 to 30mL of formalin-free composition for each IKg of specimen weight.
  • conduit of the specimen refers to veins, arteries, urinary tracts, respiratory and/or digestive tracts and any tubular component that can be channeled and injected.
  • the specimen is contacted with the formalin-free composition by dipping and injecting as described above.
  • the specimen is made viable for the first time with the method for making specimens previously described, this is repeated every 2 to 5 months for between 4 to 5 days, putting the specimen in contact with the formalin-free composition, to guarantee the conservation of the specimen. specimen indefinitely.
  • the formalin-free composition is contacted with the specimen, moistening an absorbent material with the composition, or applying the composition with an atomizer or spray.
  • the formalin-free composition to conserve and preserve tissues maintains the physical-mechanical characteristics of the tissue such as color, volume and rigidity, facilitating medical training in anatomy and surgical simulation training, since it provides an experience closer to reality. Additionally, the formalin-free composition reduces the sensitive discomfort of the users when in contact with the tissues, which is also an advantage for medical education.
  • the formalin-free composition increases the useful life of human and animal specimens used in anatomy and surgical simulation.
  • tissue preservation methods that preserve the specimen for around 6 months, with the use of the developed composition, the preservation exceeds 5 years, avoiding the need to use other alternatives such as the implementation of live animals for medical training or artificial surgical simulators with low anatomical similarity.
  • Example 1 Process to obtain a formalin-free composition
  • a previously fixed 78-kilogram human specimen was immersed in 400 liters of the formaldehyde-free composition Al described in Example 1 for 15 days. Subsequently, 2000 mL of the formalin-free composition was injected into the specimen using the right femoral artery, a branch of the right external iliac artery, as the access route.
  • the viability specimen with the formaldehyde-free composition Al has been kept preserved since the first viability by repeating the method every 3 months, contacting the specimen with the formaldehyde-free composition by means of immersion or spray mist, moistening the specimen.
  • the viability method was repeated for a period of 5 years and it is observed that the specimen maintains its characteristics and continues to be suitable for dissection in anatomy education and for medical training in surgical simulation, especially laparoscopy.
  • specimens A and B are characterized by an improvement in joint mobility with respect to specimen C, as well as a rehabilitation of the elasticity of the tissues (FIG. 3), which in the case of experimentation for surgical simulation in laparoscopy, it allows obtaining an adequate pneumoperitoneum at pressures between 5 and 7 mmHg

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Dentistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Environmental Sciences (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

La présente invention concerne une composition exempte de formol pour conserver et rendre viables des tissus. Ladite composition comprend entre 20 à 30% v/v d'un agent hydratant, entre 15 à 25% v/v d'un agent conservateur, entre 8 à 14% v/v d'un agent sporicide et fongicide, entre 4 à 10% p/v d'un agent bactériostatique, entre 0,1 à 5% v/v d'un agent répulsif et un milieux aqueux. La composition exempte de formol ne déforme pas les caractéristiques mécaniques du tissu, diminue les désagréments sensitifs des utilisateurs lors du contact avec les tissus, augmente la durée de vie des tissus et évite l'utilisation de réactifs cancérigènes. En outre, le développement fait référence à un processus visant à obtenir la composition exempte de formol et à un procédé de viabilisation de spécimens consistant à mettre en contact le spécimen avec la composition exempte de formol.
PCT/IB2021/060372 2020-11-09 2021-11-09 Composition exempte de formol pour conserver et rendre viables des tissus WO2022079702A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CONC2022/0007327A CO2022007327A2 (es) 2020-11-09 2022-05-27 Composición libre de formol para conservar y viabilizar tejidos

Applications Claiming Priority (2)

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US202063111392P 2020-11-09 2020-11-09
US63/111,392 2020-11-09

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WO2022079702A1 true WO2022079702A1 (fr) 2022-04-21

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WO (1) WO2022079702A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015138479A1 (fr) * 2014-03-10 2015-09-17 The Trustees Of Columbia University In The City Of New York Compositions antimicrobiennes pour végétaux

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015138479A1 (fr) * 2014-03-10 2015-09-17 The Trustees Of Columbia University In The City Of New York Compositions antimicrobiennes pour végétaux

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
HAMMER N ET AL.: "Ethanol-Glycerin Fixation with Thymol Conservation: A Potential Alternative to Formaldehyde and Phenol Embalming", ANAT SCI EDUC, vol. 5, 2012, pages 225 - 233, XP055934091, Retrieved from the Internet <URL:https://doi.org/10.1002/ase.1270> *
HAMMER N ET AL.: "Substitution of formaldehyde in gross anatomy is possible [ On line", J NATL CANCER INS T, vol. 103, 2011, pages 610 - 611, XP055934097, [retrieved on 20210114] *
RAHMAN MD ET AL.: "Alcoholic fixation over formalin fixation: A new, safer option for morphologic and molecular analysis of tissues", SAUDI JOURNAL OF BIOLOGICAL SCIENCES, vol. 29, no. 1, January 2022 (2022-01-01), pages 175 - 182, XP055934100, DOI: https://doi.org/10.1016/j.sjbs. 2021.08.07 5 *
TAMAYO-ARANGO L ET AL.: "Preservation of Animal Cadavers with Formaldehyde-Free Solution for Gross Anatomy", JOURNAL OF MORPHOLOGICAL SCIENCES, vol. 35, no. 02, 2018, pages 136 - 141, XP055934094, DOI: 10.1055/S-0038-1669434 *

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