EP2079787A1 - Réticulation de mousses de kératine s-sulfonée - Google Patents

Réticulation de mousses de kératine s-sulfonée

Info

Publication number
EP2079787A1
EP2079787A1 EP07817933A EP07817933A EP2079787A1 EP 2079787 A1 EP2079787 A1 EP 2079787A1 EP 07817933 A EP07817933 A EP 07817933A EP 07817933 A EP07817933 A EP 07817933A EP 2079787 A1 EP2079787 A1 EP 2079787A1
Authority
EP
European Patent Office
Prior art keywords
keratin
foam
cross
linked
foams
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP07817933A
Other languages
German (de)
English (en)
Inventor
Jens Hoeg Truelsen
Pia Nielsen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Coloplast AS
Original Assignee
Coloplast AS
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 Coloplast AS filed Critical Coloplast AS
Publication of EP2079787A1 publication Critical patent/EP2079787A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/36—After-treatment
    • 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
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
    • 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
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08H—DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS
    • C08H1/00—Macromolecular products derived from proteins
    • C08H1/06—Macromolecular products derived from proteins derived from horn, hoofs, hair, skin or leather
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2389/00—Characterised by the use of proteins; Derivatives thereof
    • C08J2389/04—Products derived from waste materials, e.g. horn, hoof or hair

Definitions

  • the present invention relates to a process for cross-linking of highly porous foams of S- sulfonated keratin (in particular scaffolds), and to highly porous, cross-linked keratin foams (e.g. scaffolds) as such.
  • Such scaffolds (foams) are particularly useful for wound care applications.
  • Foams in the form of scaffolds are guiding structures used for wound care applications resulting in development of new tissue. They are usually made up of biomaterials and are added to tissue to guide the organization, growth and differentiation of cells in the process of forming functional tissue.
  • scaffolds must meet some specific requirements. A high porosity and an adequate pore size are necessary to facilitate cell seeding and diffusion throughout the whole structure of both cells and nutrients. Biodegradability is essential since scaffolds need to be absorbed by the surrounding tissues without the necessity of a surgical removal. The rate at which degradation occurs has to coincide as much as possible with the rate of tissue formation : this means that while cells are fabricating their own natural matrix structure around themselves, the scaffold is able to provide structural integrity within the body and eventually it will break down leaving the neotissue, newly formed tissue which will take over the mechanical load. Injectability is also important for clinical uses.
  • scaffolds are those manufactured from natural or synthetic polymers with good biocompatibility and biodegradability. Such materials are, e.g., gelatin, fibrin, hyalouronic acid, collagen, chitin, chitosan, keratin, alginate, poly(L-lactic acid) (PLLA), poly(D/L-lactic acid) (PDLLA), and poly(lactic-co-glycolic acid) (PLGA).
  • PLLA poly(L-lactic acid)
  • PLLA poly(D/L-lactic acid)
  • PLGA poly(lactic-co-glycolic acid)
  • wound dressings e.g. scaffolds
  • wound dressings e.g. scaffolds
  • Soft, flexible, absorbent and coherent keratin foams are in general needed for ensuring a comfortable and proper treatment of difficult-to-heal wounds and as releasing media for active ingredients such as growth factors.
  • the present invention relates to a process for cross-linking of a highly porous foam of S-sulfonated keratin, cf. claim 1.
  • the invention further relates to a cross-linked keratin foam, cf. claim 7, various uses of such a cross-linked keratin foam, cf. claims 11 and 12, and to a method of supporting cell adhesion and/or the in-growth for regeneration of tissue utilizing said keratin foam, cf. claim 13.
  • foam is intended to encompass light-weight structures conventionally known as foams, sponges, scaffolds, or "porous structures”.
  • the term "scaffold” is intended to mean porous structures into which cells may be incorporated (in-growth).
  • Scaffolds serve at least one of the following purposes:
  • the present invention relates to a process for cross-linking of a highly porous foam of S-sulfonated keratin, said process comprising the steps of
  • a highly porous foam of S-sulfonated keratin is provided.
  • the highly porous foam is prepared by formation of a solution having a content of S-sulfonated keratin of at the most 30 img/mL, i.e. corresponding to a final foam having a density of at the most
  • the S-sulfonated keratin may be obtained via well-known procedures, e.g. as described in WO 03/011894 Al.
  • the solvent in which the S-sulfonated keratin is dissolved is typically an aqueous solvent, i.e. a solvent comprising at least 85% (w/w) of water.
  • the S-sulfonated keratin is soluble only as the salt, which can be prepared by the addition of a base to the S-sulfonated keratin.
  • the solvent is an aqueous solvent comprising at least 96% (w/w) of water and up to 4% (w/w) of organic constituents or salts.
  • the up to 4% (w/w) of non-aqueous constituent comprises at least one organic constituent selected from porosity improvers such as tert-butanol and softeners such as glycerol, low molecular weight polyethyleneglycol (i.e. liquid polyethyleneglycol), liquid Pluronic®, Tween® or other chemicals that may have softening properties.
  • the solvent consists of at least 97% of water in admixture with tert-butanol and glycerol.
  • the solvent is water comprising a strong base.
  • the base is typically present in an amount providing a pH of about 9 to 10. This may be obtained by adding 1 imL of a 1 M NaOH (or similar, e.g. 1 M KOH or ammonia) per gram of the S-sulfonated keratin.
  • the foam may be prepared as described in WO 03/018673 Al.
  • the concentration of the S-sulfonated keratin is typically at the most 30 img/mL, e.g. in the range of 5-30 img/mL, such as in the range of 10-25 img/mL.
  • the solution is subsequently cast onto a flat surface or in a mould so as to obtain a foam of the intended shape or a foam which can be later cut in suitable pieces.
  • the aqueous solution is subsequently frozen so as to obtain a frozen solution of S-sulfonated keratin; and the frozen solution is subsequently freeze-dried so as to obtain a highly porous foam S- sulfonated keratin.
  • the foam is a scaffold adapted for wound care applications.
  • step (a) the highly porous keratin foam is cross-linked by means of a reductant.
  • the treatment with a suitable reductant causes the sulfonate groups to be removed from the S-sulfonated cystine groups of the keratin whereby two cystine groups react and form a disulfide linkage.
  • Suitable reductants are those selected from ammonium thioglycolate, tributylphosphine, and triscarboxyethylphosphine hydrochloride. Two or more reductants may be used in combination, if desired.
  • step (b) An important feature of step (b) is that the reductant is dissolved in a solvent comprising at least 85% of a Ci- 6 -alcohol.
  • a solvent comprising at least 85% of a Ci- 6 -alcohol.
  • Ci- 6 -alcohols comprise methanol, ethanol, 1-propanol, 2-propanol (iso-propanol), 1-butanol, 2-butanol, 2-methyl-l-butanol (iso-butanol), 2-methyl-2-butanol (tert-butanol), 1-pentanol, 1-hexanol, etc.
  • the currently most preferred examples of Ci- 6 -alcohols are ethanol and 2- propanol, in particular ethanol.
  • the solvent comprises at least 90%, such as at least 95% of the Ci- 6 -alcohol.
  • the reductant is typically used in an amount of 0.1-40 mmol per gram of the S-sulfonated keratin. More particular, an amount corresponding to 0.5-20 mmol per gram, such as 0.8-9 mmol per gram, is used.
  • the reductant is allowed to interact with the foam of the S-sulfonated keratin for a period sufficient to obtain the desired degree of cross-linking.
  • the cross-linking is allowed to take place somewhere between 1 minute to 24 hours, e.g. from 5 minutes to 12 hours. It will be evident for the skilled person, that the period can be reduced if the cross-linking takes place at elevated temperature, e.g. 25-50 0 C, and that prolonged reaction is necessary if the reaction takes place at a low temperature, e.g. -10 to 10 0 C.
  • a specific, illustrative example of a reductant solution is 0.003-0.05 M ammonium thioglycolate in 92-99.8% ethanol.
  • step (b) the cross-linked keratin foam is separated from the reductant solution.
  • the reductant solution is typically removed by passive dripping off, by suction (moderate vacuum), or by compression of the foam.
  • the process comprises the further step (d) of washing the cross- linked keratin foam with a solution comprising at least 90% of a Ci- 6 -alcohol.
  • the solution comprises at least 95% of the Ci- 6 -alcohol.
  • Ci- 6 -alcohol is as described above under step (b), and also in step (d), the currently most preferred examples of Ci- 6 -alcohols are ethanol and 2-propanol, in particular ethanol.
  • the washing may be conducted by flushing, or by dipping, typically in 1 to 3 steps.
  • the void space of the foam may be unoccupied so as to allow cell adhesion and/or in-growth for regeneration of tissue.
  • the pores of the material are at least partly occupied by particles or a component from the extracellular matrix or the foam is coated with particles or a component from the extracellular matrix. Such particles and components may facilitate the cell adhesion and/or in-growth for regeneration of tissue.
  • components from the extracellular matrix are chondroitin sulfate, hyaluronan, hyaluronic acid, heparin sulfate, heparan sulfate, dermatan sulfate, growth factors, fibrin, fibronectin, elastin, collagen, gelatin, and aggrecan.
  • the process of the invention also encompasses a variant wherein particles of the extracellular matrix or components from the extracellular matrix are dispersed or dissolved in the solution of the S-sulfonated keratin used in step (a) before the aqueous solution (dispersion) is frozen and freeze-dried.
  • the process invention also encompasses the more specific variant wherein the components from the extracellular matrix are dissolved in a suitable solvent and then added to the solution of the S-sulfonated keratin. By mixing with the solvent of the S-sulfonated keratin, the components from the extracellular matrix will most likely precipitate so as to form a dispersion.
  • the process of the invention encompasses a variant wherein the cross-linked keratin foam obtained in step (c), in a subsequent step (e.g. as an alternative or supplement to step (d)), is immersed in a solution of glucosaminoglycan (e.g. hyaluronan) and subsequently freeze-dried again.
  • glucosaminoglycan e.g. hyaluronan
  • the foams (in particular scaffolds) prepared according to the process of the present invention are particularly useful for tissue regeneration. Besides the use of the foams (scaffolds) for tissue regeneration within wound care applications it may also be used within continence care and ostomy care.
  • the foams have a multitude of uses within the field of medicine, healthcare, surgery, dentistry, etc., in particular uses where a biodegradable polymer is required, e.g. for wound dressings, scaffolds for cell attachment and in-growth for regeneration of tissue, hernia-mesh, cartilage, ligaments, implants, etc.
  • the present invention also relates to a cross-linked keratin foam as defined herein for use in therapy, dentistry or surgery.
  • the invention also relates to the use of a cross-linked keratin foam as defined herein for the preparation of a scaffold for supporting cell adhesion or the in-growth for regeneration of tissue, and to the use of a cross-linked keratin foam as defined herein for the preparation of a scaffold adapted for wound care applications.
  • the invention further relates to a method of supporting cell adhesion and/or the in-growth for regeneration of tissue, the method comprising the step of contacting a cross-linked keratin foam as defined herein with said tissue, e.g. soft tissue such as skin, ligament, tendon, cartilage, and bone.
  • tissue e.g. soft tissue such as skin, ligament, tendon, cartilage, and bone.
  • S-sulfonated proteins may be used in a similar manner, i.e. by utilizing approximately the same means, to produce alternative, highly porous foams.
  • Keratin scaffolds were prepared form 2% (w/w), 1.5% (w/w), 1.4% (w/w) and 0.8% (w/w) S-sulfonated keratin solutions.
  • Table 1 shows the chemical compositions and the concentration of each component in the stock solutions prior to the lyophilisation.
  • a solution comprising 0.25 M ammonium thioglycolate and 0.1 M potassium phosphate buffer was prepared in a 1 L measuring flask. Keratin scaffolds prepared as described in Example 1 (one keratin scaffold from sample 1 and one keratin scaffold from sample 6 (see Table I)) were cross-linked using following procedure.
  • the keratin scaffold was immersed in 50 imL of the 0.25 M ammonium thioglycolate solution for 30 minutes.
  • the cross-linked scaffold was afterwards washed 3 times by immersing in 50 imL demineralised water for 10 min.
  • the cross-linked and washed keratin scaffolds was finally lyophilised.
  • the cross-linked keratin scaffolds obtained via this route were collapsed, had a significant reduced porosity and a highly increased stiffness making them less usable for would care applications.
  • the scaffolds prepared as described in Example 1 were cross- linked by immersion in 50 imL of the ammonium thioglycolate solution for 30 minutes (see Table 2).
  • the cross-linked scaffolds were afterwards washed 3 times with 96% ethanol by immersing in 96% ethanol for 10 minutes per wash. The ethanol is exchanged between each wash.
  • the cross-linked and washed keratin scaffolds were afterwards dried in a vacuum oven over night at room temperature.
  • the obtained cross-linked scaffolds were highly porous, flexible and water absorbent making them attractive for wound care application.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Transplantation (AREA)
  • Epidemiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Animal Behavior & Ethology (AREA)
  • Oral & Maxillofacial Surgery (AREA)
  • Dermatology (AREA)
  • Biochemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Materials For Medical Uses (AREA)
  • Manufacture Of Porous Articles, And Recovery And Treatment Of Waste Products (AREA)

Abstract

La présente invention concerne un procédé pour réticuler des mousses hautement poreuses de kératine S-sulfonée, et des mousses de kératine réticulées, hautement poreuses (par exemple des scaffolds) en tant que tels. De tels scaffolds (mousses) sont particulièrement utiles pour soigner les plaies. De telles mousses de kératine poreuses peuvent être obtenues par en remplaçant la solution aqueuse du réducteur comme milieu de réticulation par une solution alcoolique du même réducteur, et en remplaçant les procédures de lavage aqueux par une procédure similaire de lavage avec un alcool.
EP07817933A 2006-10-02 2007-10-02 Réticulation de mousses de kératine s-sulfonée Withdrawn EP2079787A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA200601269 2006-10-02
PCT/DK2007/050134 WO2008040357A1 (fr) 2006-10-02 2007-10-02 Réticulation de mousses de kératine s-sulfonée

Publications (1)

Publication Number Publication Date
EP2079787A1 true EP2079787A1 (fr) 2009-07-22

Family

ID=37946243

Family Applications (1)

Application Number Title Priority Date Filing Date
EP07817933A Withdrawn EP2079787A1 (fr) 2006-10-02 2007-10-02 Réticulation de mousses de kératine s-sulfonée

Country Status (4)

Country Link
EP (1) EP2079787A1 (fr)
AU (1) AU2007304585A1 (fr)
CA (1) CA2665253A1 (fr)
WO (1) WO2008040357A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2430037A4 (fr) 2009-05-13 2013-07-17 Keraplast Tech Ltd Matériaux biopolymères
WO2014112950A1 (fr) * 2013-01-18 2014-07-24 Nanyang Technological University Méthode de préparation d'un biomatériau à base de kératine et biomatériau à base de kératine formé correspondant

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6110487A (en) * 1997-11-26 2000-08-29 Keraplast Technologies Ltd. Method of making porous keratin scaffolds and products of same
WO2003018673A1 (fr) * 2001-08-31 2003-03-06 Keratec Limited Production de materiaux biopolymeres sous forme de films, de fibres, de mousses ou d'adhesifs, a partir de derives de keratine s-sulfonee
JP4551761B2 (ja) * 2002-06-10 2010-09-29 ケラテク リミテッド ケラチン由来の整形外科材料
WO2005058380A1 (fr) * 2003-12-19 2005-06-30 Keratec Limited Produits de soin pour les plaies contenant de la keratine

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2008040357A1 *

Also Published As

Publication number Publication date
AU2007304585A1 (en) 2008-04-10
WO2008040357A1 (fr) 2008-04-10
CA2665253A1 (fr) 2008-04-10

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