WO2002072113A1 - Wound healing using fibroblasts - Google Patents

Wound healing using fibroblasts Download PDF

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Publication number
WO2002072113A1
WO2002072113A1 PCT/GB2002/001061 GB0201061W WO02072113A1 WO 2002072113 A1 WO2002072113 A1 WO 2002072113A1 GB 0201061 W GB0201061 W GB 0201061W WO 02072113 A1 WO02072113 A1 WO 02072113A1
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WIPO (PCT)
Prior art keywords
composition according
pharmaceutical composition
fibroblasts
cells
skin wound
Prior art date
Application number
PCT/GB2002/001061
Other languages
French (fr)
Inventor
Michael David Leek
Paul David Kemp
Original Assignee
Intercytex Limited
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
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Publication of WO2002072113A1 publication Critical patent/WO2002072113A1/en

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    • 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
    • A61K35/14—Blood; Artificial blood
    • A61K35/16—Blood plasma; Blood serum

Definitions

  • the invention relates to scarring as a result of wound healing.
  • the invention relates to a composition, use of the composition and method of treatment of fibrosis and scarring during wound healing.
  • Fibrosis and the subsequent scarring as a result of wound healing is a serious clinical problem.
  • Tissue that has healed inappropriately can lead to disfigurement, and the accompanying psychological effect on the person.
  • scarring can lead to an impairment function of the tissues. This is particularly important for organ tissues or other non-dermal tissues whose specialist function can be disrupted or impaired by scarring. Scarring as a result of surgery for tumour excision, for example.
  • the initial phase concerns the formation of a blood clot with entrapped platelets.
  • the platelets release a mix of pro-inflammatory factors into the clot.
  • various factors released by the forming clot activate endothelial cells in the surrounding blood vessels, making them "sticky" to particular circulating white blood cells.
  • circulating blood cells become attached to the endothelium and then are attracted into the wound.
  • the first of these cells release cytotoxic substances to reduce infection.
  • the second type of cells (monocyte/macrophage) then engulf the neutrophils and any cellular or matrix debris.
  • fibroblasts enter the wound from the surrounding tissues and begin the rebuilding process.
  • blood vessels grow into the wound. On some occasions the growth can be too vigorous and result in the reddening of the wound.
  • connective tissue cells are suitable cells. These include cell types such as fibroblasts, myoblasts or smooth muscle cells.
  • composition comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix-forming material, and the composition is substantially free of other cell types, for use in the reduction of fibrosis and scar tissue during skin wound healing.
  • the fibroblasts comprise at least 90%, preferably at least 91, 92, 93, 94, 95, 96, 97, 98 or 99%, of the cells.
  • the composition may be completely free of cell types other than fibroblasts.
  • the composition at the time of incorporation of living cells may be free, or substantially free, of pre-formed matrix material.
  • Matrix-forming material exists in a pre-matrix constitution in the composition but has the ability to form a scaffold or matrix around the cells in the composition.
  • the matrix-forming material will solidify in use so that the composition forms a construct comprising a scaffold or matrix containing the appropriate cells.
  • the scaffold or matrix may be formed around the cells either in situ or prior to application of the composition to the wound.
  • the cells of the invention can be provided in a composition
  • a composition comprising a matrix-forming material that carries the cells to the wound site, and immobilises the cells at the site.
  • matrix-forming material can be selected from fibrinogen/thrombin mixture, for example.
  • the composition may be formulated with a suitable carrier to aid delivery to the wound site such as hyaluronic acid.
  • thrombin converts fibrinogen to fibrin which forms a matrix, immobilising the cells at the required site.
  • the composition can be in a suitable form for storage prior to use. Further, the fibrinogen may be converted prior to administration or after administration to the wound site.
  • the composition when the composition is required it can be prepared using commonly known, pharmaceutically acceptable methods.
  • fibrin may be used to form the matrix. This could be applied directly to the wound or in a suitable carrier which would deliver the composition to the wound site.
  • composition of cells and matrix-forming material can be prepared in a pharmaceutically acceptable carrier, diluent or excipient prior to administration.
  • composition can be formulated into a pharmaceutically acceptable form but then stored.
  • formulation can be made ready by the addition of a suitable diluent to form the required solution.
  • the fibroblasts may be mammalian, preferably human.
  • the invention provides that the cells could be allogeneic cells, i.e. the cells administered to a patient would be from a donor.
  • compositions comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix- forming material, and the composition is substantially free of other cell types, formulated with a pharmaceutically acceptable carrier, diluent or excipient.
  • the constituents of the pharmaceutical composition may include those as stated above for the composition.
  • the pharmaceutical composition as a formulation can be suitable for either topical delivery or parenteral delivery.
  • the formulation could be in the form of an ointment or paste for applying to the external surface of the wound.
  • parenteral delivery the formulation could be in the form suitable for injection at the wound site.
  • Such formulations would include solutions or suspensions, with or without a carrier such as a microsphere or microcapsule .
  • the pharmaceutical composition may be in the form of an ointment or paste for applying to the external surface of the wound. Also provided is the use of a composition or a pharmaceutical composition according to the invention in the reduction of fibrosis and scarring as a result of skin wound healing.
  • compositions or a pharmaceutical composition according to the invention in the manufacture of a medicament for the reduction of fibrosis and scarring during skin wound healing.
  • the invention encompasses a method of reducing fibrosis and scarring during skin wound healing comprising the administration to the patient of an effective amount of a composition or a pharmaceutical composition according to the invention.
  • composition or pharmaceutical composition of the invention may be used within a number of hours after a wound has formed, for example immediately after a wound has formed, although it may be preferable to administer it within a shorter time.
  • the composition or pharmaceutical composition may be used before the inflammatory phase of the skin wound. This has the effect of minimising fibrosis and scarring.
  • the composition or pharmaceutical composition may be administered 2-48 hours, preferably 2-36 hours, after wounding.
  • composition or pharmaceutical composition may be administered before inflammation of the wound site.
  • it may be necessary to administer it over a sustained period of time.
  • the method is applicable when the skin wound is an acute skin wound or a chronic skin wound.
  • the invention also provides for a method of decreasing the wound healing time comprising administering an effective amount of a composition or pharmaceutical composition according to the invention.
  • Porcine fibroblasts for the fibrin delivery vehicle were prepared as follows: a full thickness skin biopsy was taken and placed into a small volume of supplemented DMEM. Using fine forceps and scalpel the dermis was dissected away from fatty tissue and placed into a falcon tube containing lxPBS. The biopsy was washed by shaking vigorously - this step was repeated 3 times. Following the washes, the biopsy was finely chopped using a scalpel and the resulting tissue transferred into a falcon tube containing a 1 x Collagenase solution (lOOU/ml) . The tissue was continually agitated at 37°C for 90 minutes then allowed to settle.
  • the supernatant was removed, placed into a fresh falcon tube and centrifuged at lOOOrpm for 5 minutes.
  • the pellet was re-suspended in 2ml of supplemented DMEM.
  • a cell count was performed to assess number.
  • Fresh collagenase solution was added to the digesting tissue and the above procedure repeated. The procedure was continued until the fraction yield diminished to approximately 10% of the highest fraction yield (typically 3-5 times) . All fractions were combined and plated out at the desired density (10, 000 cells/cm 2 ) . The cells were expanded and maintained until the required number achieved. The dissociation and expansion was typically performed 2 weeks prior to application of the construct to the porcine wounds.
  • Porcine fibroblasts were incorporated into the construct as follows: Each construct was cast with 500,000 cells in a final volume of 500ml. The desired number of cells were resuspended in 450ml of a 7.5mg/ml fibrinogen (typically purified from cryoprecipitate by methods known to those skilled in the art) and supplemented DMEM .
  • fibrinogen could be purchased from Sigma.
  • Thrombin Sigma: 50ml of (lOOU/ml) ] was added to a 48 well plate, and shaken gently to ensure an even distribution of thrombin over the base of the plate. The fibrinogen/cell suspension mixture was added to the thrombin and clotting occurred within seconds.
  • 0.5ml of DMEM was added to the clot and allowed to mature for 24 hours.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Cell Biology (AREA)
  • Hematology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention relates to scarring as a result of wound healing. In particular, the invention relates to a composition and a pharmaceutical composition comprising cells capable of reducing an inflammation response caused by wounding and a matrix-forming material, use of these compositions for reducing fibrosis and scarring, and a method of treatment of fibrosis and scarring during wound healing.

Description

WOUND HEALING USING FIBROBLASTS
The invention relates to scarring as a result of wound healing. In particular, the invention relates to a composition, use of the composition and method of treatment of fibrosis and scarring during wound healing.
Wounds often heal with the production of a scar, which provides mechanical strength to a healed wound but can be unsightly. However, the exact mechanism of scar formation is not known. It has been proposed that it is likely to be a consequence of the body's need, in historical terms, to have a robust inflammatory response to any open wound in order to combat infection. The inflammatory response has the "side effect" of over stimulation of the synthesis of a fibre matrix (fibrosis) . This inappropriate laying down of such fibres results in the appearance of a scar.
Fibrosis and the subsequent scarring as a result of wound healing is a serious clinical problem. Tissue that has healed inappropriately can lead to disfigurement, and the accompanying psychological effect on the person. Further, scarring can lead to an impairment function of the tissues. This is particularly important for organ tissues or other non-dermal tissues whose specialist function can be disrupted or impaired by scarring. Scarring as a result of surgery for tumour excision, for example.
Following wounding, the normal cellular healing response is triggered. This process involves a series of processes.
The initial phase concerns the formation of a blood clot with entrapped platelets. The platelets, in turn, release a mix of pro-inflammatory factors into the clot. In the second phase, various factors released by the forming clot activate endothelial cells in the surrounding blood vessels, making them "sticky" to particular circulating white blood cells.
In the third phase, particular circulating blood cells become attached to the endothelium and then are attracted into the wound. The first of these cells (neutrophils) release cytotoxic substances to reduce infection. The second type of cells (monocyte/macrophage) then engulf the neutrophils and any cellular or matrix debris.
In a fourth phase, fibroblasts enter the wound from the surrounding tissues and begin the rebuilding process.
In a fifth phase, blood vessels grow into the wound. On some occasions the growth can be too vigorous and result in the reddening of the wound.
Although described here as five distinct phases, it will be appreciated that these phases are not always distinct, and that each phase may "blend" with the next.
It is, however, during the first and second phases when the triggers to inflammation and then the inflammatory factors themselves are released. It is during this time that it is thought that the triggers that cause fibrosis and scarring are released.
Currently, there is only one treatment approved and available to reduce scarring, that of silicone gel. However, the mode of action is unknown. Furthermore, the normal management of wounds is concerned with the initial closure of the wound (for example by sutures) , and the prevention of infection and dessication of the wound with a suitable dressing. Little is done to prevent scarring in most cases, although good standards of care can minimise fibrosis and scarring.
Surprisingly, it has now been found that the introduction of cells capable of reducing the inflammatory response caused by wounding reduces fibrosis and scarring. The cells are introduced at a wound site early in the healing cascade, and hence interfere with the subsequent phases of the cascade.
Further, it has been found that connective tissue cells are suitable cells. These include cell types such as fibroblasts, myoblasts or smooth muscle cells.
Thus according to the invention there is provided a composition comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix-forming material, and the composition is substantially free of other cell types, for use in the reduction of fibrosis and scar tissue during skin wound healing.
By "substantially free of other cell types", it is meant that the fibroblasts comprise at least 90%, preferably at least 91, 92, 93, 94, 95, 96, 97, 98 or 99%, of the cells. Alternatively, the composition may be completely free of cell types other than fibroblasts.
The composition at the time of incorporation of living cells may be free, or substantially free, of pre-formed matrix material. Matrix-forming material exists in a pre-matrix constitution in the composition but has the ability to form a scaffold or matrix around the cells in the composition. For example, if the composition is in the form of a gel, the matrix-forming material will solidify in use so that the composition forms a construct comprising a scaffold or matrix containing the appropriate cells. This differs from prior art where cells are incorporated into an exiting, preformed matrix or scaffold. In the present invention, the scaffold or matrix may be formed around the cells either in situ or prior to application of the composition to the wound.
Thus the cells of the invention can be provided in a composition comprising a matrix-forming material that carries the cells to the wound site, and immobilises the cells at the site. Such matrix-forming material can be selected from fibrinogen/thrombin mixture, for example. The composition may be formulated with a suitable carrier to aid delivery to the wound site such as hyaluronic acid.
If a fibrinogen/thrombin mixture is used, at the wound site, thrombin converts fibrinogen to fibrin which forms a matrix, immobilising the cells at the required site. This has the advantage that the composition can be in a suitable form for storage prior to use. Further, the fibrinogen may be converted prior to administration or after administration to the wound site. In addition, when the composition is required it can be prepared using commonly known, pharmaceutically acceptable methods.
Further, fibrin may be used to form the matrix. This could be applied directly to the wound or in a suitable carrier which would deliver the composition to the wound site.
The composition of cells and matrix-forming material can be prepared in a pharmaceutically acceptable carrier, diluent or excipient prior to administration.
Alternatively, the composition can be formulated into a pharmaceutically acceptable form but then stored. When required, the formulation can be made ready by the addition of a suitable diluent to form the required solution.
The fibroblasts may be mammalian, preferably human. The invention provides that the cells could be allogeneic cells, i.e. the cells administered to a patient would be from a donor.
Further provided according to the invention is a pharmaceutical composition comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix- forming material, and the composition is substantially free of other cell types, formulated with a pharmaceutically acceptable carrier, diluent or excipient.
The constituents of the pharmaceutical composition may include those as stated above for the composition.
The pharmaceutical composition as a formulation can be suitable for either topical delivery or parenteral delivery. For topical delivery, the formulation could be in the form of an ointment or paste for applying to the external surface of the wound. For parenteral delivery, the formulation could be in the form suitable for injection at the wound site. Such formulations would include solutions or suspensions, with or without a carrier such as a microsphere or microcapsule .
The pharmaceutical composition may be in the form of an ointment or paste for applying to the external surface of the wound. Also provided is the use of a composition or a pharmaceutical composition according to the invention in the reduction of fibrosis and scarring as a result of skin wound healing.
In another aspect of the invention, there is provided the use of a composition or a pharmaceutical composition according to the invention in the manufacture of a medicament for the reduction of fibrosis and scarring during skin wound healing.
The invention encompasses a method of reducing fibrosis and scarring during skin wound healing comprising the administration to the patient of an effective amount of a composition or a pharmaceutical composition according to the invention.
It is envisaged that the composition or pharmaceutical composition of the invention may be used within a number of hours after a wound has formed, for example immediately after a wound has formed, although it may be preferable to administer it within a shorter time. Thus the composition or pharmaceutical composition may be used before the inflammatory phase of the skin wound. This has the effect of minimising fibrosis and scarring. The composition or pharmaceutical composition may be administered 2-48 hours, preferably 2-36 hours, after wounding.
Preferably the composition or pharmaceutical composition may be administered before inflammation of the wound site. In addition, it may be necessary to administer it over a sustained period of time.
The method is applicable when the skin wound is an acute skin wound or a chronic skin wound. The invention also provides for a method of decreasing the wound healing time comprising administering an effective amount of a composition or pharmaceutical composition according to the invention.
Experimental example:
Porcine fibroblasts for the fibrin delivery vehicle (construct) were prepared as follows: a full thickness skin biopsy was taken and placed into a small volume of supplemented DMEM. Using fine forceps and scalpel the dermis was dissected away from fatty tissue and placed into a falcon tube containing lxPBS. The biopsy was washed by shaking vigorously - this step was repeated 3 times. Following the washes, the biopsy was finely chopped using a scalpel and the resulting tissue transferred into a falcon tube containing a 1 x Collagenase solution (lOOU/ml) . The tissue was continually agitated at 37°C for 90 minutes then allowed to settle. The supernatant was removed, placed into a fresh falcon tube and centrifuged at lOOOrpm for 5 minutes. The pellet was re-suspended in 2ml of supplemented DMEM. A cell count was performed to assess number. Fresh collagenase solution was added to the digesting tissue and the above procedure repeated. The procedure was continued until the fraction yield diminished to approximately 10% of the highest fraction yield (typically 3-5 times) . All fractions were combined and plated out at the desired density (10, 000 cells/cm2) . The cells were expanded and maintained until the required number achieved. The dissociation and expansion was typically performed 2 weeks prior to application of the construct to the porcine wounds.
Porcine fibroblasts were incorporated into the construct as follows: Each construct was cast with 500,000 cells in a final volume of 500ml. The desired number of cells were resuspended in 450ml of a 7.5mg/ml fibrinogen (typically purified from cryoprecipitate by methods known to those skilled in the art) and supplemented DMEM . In an alternative example, fibrinogen could be purchased from Sigma. Thrombin [Sigma: 50ml of (lOOU/ml) ] was added to a 48 well plate, and shaken gently to ensure an even distribution of thrombin over the base of the plate. The fibrinogen/cell suspension mixture was added to the thrombin and clotting occurred within seconds. 0.5ml of DMEM was added to the clot and allowed to mature for 24 hours.
Both excisional and incisional wounds (in Yorkshire pigs) made by standard methods to those skilled in the art were treated with the construct on Day 0 (day of wound) and day 4 (4 days post-wound) . The wounds containing the constructs were covered by an occlusive dressing. Treatments were made in the following groups:
day 0: FB on fibrin matrix + epidermal graft (excisional)
FB on fibrin matrix - epidermal graft (excisional) FB on fibrin matrix into an incisional wound
day 4: FB on fibrin delivery system + epidermal graft (excisional)
FB on fibrin delivery system - epidermal graft (excisional) FB on fibrin matrix into an incisional wound
At day 4, 7, 21 and 56 wounds are macroscopically evaluated and punch biopsies are taken for histological evaluation. At 21 and 56 days after surgery wound contraction was determined.

Claims

Claims
1. A composition comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix-forming material, and the composition is substantially free of other cell types, for use in the reduction of fibrosis and scar tissue during skin wound healing.
2. The composition according to claim 1, wherein the fibroblasts comprise at least 90% of the cells.
3. The composition according to either one of claim 1 or claim 2, wherein the composition is free of cell types other than fibroblasts.
4. The composition according to any one of the preceding claims, wherein the composition is free of pre-formed matrix material.
5. The composition according to any one of the preceding claims, wherein the matrix-forming material is fibrinogen/thrombin.
6. The composition according to any one of the preceding claims, wherein the fibroblasts are mammalian, preferably human .
7. The composition according to any one of the preceding claims, wherein the fibroblasts are allogeneic.
8. The composition according to any one of the preceding claims, for use before the inflammatory phase of the skin wound.
9. A pharmaceutical composition comprising cells capable of reducing an inflammatory response caused by skin wounding and a cell delivery vehicle capable of delivering and maintaining the cells within a skin wound, wherein the cells are fibroblasts, the cell delivery vehicle is a matrix- forming material, and the composition is substantially free of other cell types, formulated with a pharmaceutically acceptable carrier, diluent or excipient.
10. The pharmaceutical composition according to claim 9, wherein the fibroblasts comprise at least 90% of the cells.
11. The pharmaceutical composition according to either one of claim 9 or claim 10, wherein the pharmaceutical composition is free of cell types other than fibroblasts.
12. The pharmaceutical composition according to any one of claims 9-11, wherein the pharmaceutical composition is free of pre-formed matrix material.
13. The pharmaceutical composition according to any one of claims 9-12, wherein the matrix-forming material is fibrinogen/thrombin .
14. The pharmaceutical composition according to any one of claims 9-13, wherein the fibroblasts are mammalian, preferably human.
15. The pharmaceutical composition according to any one of claims 9-14, wherein the fibroblasts are allogeneic.
16. The pharmaceutical composition according to any one of claims 9-15, in the form of an ointment or paste for applying to the external surface of the wound.
17. The pharmaceutical composition according to any one of claims 9-15, in the form of solution or suspension, optionally with a carrier such as a microsphere or microcapsule, for injection at a wound site.
18. The use of a composition according to claims 1-8 or a pharmaceutical composition according to claims 9-17 in the reduction of fibrosis and scarring as a result of skin wound healing.
19. The use of a composition according to claims 1-8 or a pharmaceutical composition according to claims 9-17 in the manufacture of a medicament for the reduction of fibrosis and scarring during skin wound healing.
20. A method of reducing fibrosis and scarring during skin wound healing comprising the administration to the patient of an effective amount of a composition according to claims 1-8 or a pharmaceutical composition according to claims 9- 17.
21. The method according to claim 20, wherein the composition or pharmaceutical composition is administered immediately after wounding.
22. The method according to claim 21, wherein the composition or pharmaceutical composition is administered 2- 48 hours, preferably 2-36 hours, after wounding.
23. The method according to any one of claims 20-22, wherein the skin wound is an acute skin wound.
24. The method according to any one of claims 20-23, wherein the skin wound is a chronic skin wound.
PCT/GB2002/001061 2001-03-09 2002-03-08 Wound healing using fibroblasts WO2002072113A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB0105899.9A GB0105899D0 (en) 2001-03-09 2001-03-09 Fibrosis reduction treatment
GB0105899.9 2001-03-09

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004006942A1 (en) * 2002-07-12 2004-01-22 Boston Scientific Limited Method for sustaining direct cell delivery
WO2005079822A2 (en) * 2004-02-13 2005-09-01 Intercytex Limited Pharmaceutical composition comprising a fibrin matrix and dermal fibroblasts for the treatment of wounds
US8114670B2 (en) 2005-03-14 2012-02-14 Dfb Technology Holdings, Llc Skin equivalent culture
US9271923B2 (en) 2005-05-26 2016-03-01 Intercytex Limited Tissue repair using allogeneic dermal fibroblasts
US9844473B2 (en) 2002-10-28 2017-12-19 Smith & Nephew Plc Apparatus for aspirating, irrigating and cleansing wounds
US11298453B2 (en) 2003-10-28 2022-04-12 Smith & Nephew Plc Apparatus and method for wound cleansing with actives
US11617823B2 (en) 2004-04-27 2023-04-04 Smith & Nephew Plc Wound cleansing apparatus with stress

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DE4127570A1 (en) * 1991-08-21 1993-02-25 Battelle Institut E V Antimicrobial effects of human in-vitro keratinocyte cultures - useful in the treatment of second or third degree burns and of badly healing wounds
RU2023424C1 (en) * 1993-01-13 1994-11-30 Научно-производственный центр трансплантации и культивирования тканей Methof for curing wound
US5591444A (en) * 1995-07-28 1997-01-07 Isolagen Technologies, Inc. Use of autologous dermal fibroblasts for the repair of skin and soft tissue defects
WO1997025995A1 (en) * 1996-01-18 1997-07-24 Johnson & Johnson Consumer Products, Inc. Methods for regenerating scarless skin and compositions used therein
WO1997041208A1 (en) * 1996-04-26 1997-11-06 Case Western Reserve University Skin regeneration using mesenchymal stem cells

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DE4127570A1 (en) * 1991-08-21 1993-02-25 Battelle Institut E V Antimicrobial effects of human in-vitro keratinocyte cultures - useful in the treatment of second or third degree burns and of badly healing wounds
RU2023424C1 (en) * 1993-01-13 1994-11-30 Научно-производственный центр трансплантации и культивирования тканей Methof for curing wound
US5591444A (en) * 1995-07-28 1997-01-07 Isolagen Technologies, Inc. Use of autologous dermal fibroblasts for the repair of skin and soft tissue defects
WO1997025995A1 (en) * 1996-01-18 1997-07-24 Johnson & Johnson Consumer Products, Inc. Methods for regenerating scarless skin and compositions used therein
WO1997041208A1 (en) * 1996-04-26 1997-11-06 Case Western Reserve University Skin regeneration using mesenchymal stem cells

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004006942A1 (en) * 2002-07-12 2004-01-22 Boston Scientific Limited Method for sustaining direct cell delivery
US10842678B2 (en) 2002-10-28 2020-11-24 Smith & Nephew Plc Apparatus for aspirating, irrigating and cleansing wounds
US10278869B2 (en) 2002-10-28 2019-05-07 Smith & Nephew Plc Apparatus for aspirating, irrigating and cleansing wounds
US9844473B2 (en) 2002-10-28 2017-12-19 Smith & Nephew Plc Apparatus for aspirating, irrigating and cleansing wounds
US11298453B2 (en) 2003-10-28 2022-04-12 Smith & Nephew Plc Apparatus and method for wound cleansing with actives
US8765169B2 (en) 2004-02-13 2014-07-01 Smith & Nephew, Inc. Wound healing profile
JP2015134815A (en) * 2004-02-13 2015-07-27 スミス アンド ネフュー オルトペディクス アクチェンゲゼルシャフト Wound healing composition
AU2005215211B2 (en) * 2004-02-13 2011-11-03 Smith & Nephew Orthopaedics Ag Wound healing profile
WO2005079822A2 (en) * 2004-02-13 2005-09-01 Intercytex Limited Pharmaceutical composition comprising a fibrin matrix and dermal fibroblasts for the treatment of wounds
JP2012192206A (en) * 2004-02-13 2012-10-11 Dfb Technology Holdings Llc Wound healing composition
AU2005215213B2 (en) * 2004-02-13 2010-09-09 Smith & Nephew Orthopaedics Ag Pharmaceutical composition comprising a fibrin matrix and dermal fibroblasts for the treatment of wounds
JP2015063555A (en) * 2004-02-13 2015-04-09 スミス アンド ネフュー オルトペディクス アクチェンゲゼルシャフト Wound healing compositions
AU2005215213C1 (en) * 2004-02-13 2011-04-28 Smith & Nephew Orthopaedics Ag Pharmaceutical composition comprising a fibrin matrix and dermal fibroblasts for the treatment of wounds
US9248153B2 (en) 2004-02-13 2016-02-02 Smith & Nephew, Inc. Wound healing profile
WO2005079821A1 (en) 2004-02-13 2005-09-01 Intercytex Limited Wound healing profile
US9526746B2 (en) 2004-02-13 2016-12-27 Smith & Nephew, Inc. Wound healing composition
JP2007522195A (en) * 2004-02-13 2007-08-09 インターサイテックス リミティド Wound treatment composition
WO2005079822A3 (en) * 2004-02-13 2005-11-03 Intercytex Ltd Pharmaceutical composition comprising a fibrin matrix and dermal fibroblasts for the treatment of wounds
US11617823B2 (en) 2004-04-27 2023-04-04 Smith & Nephew Plc Wound cleansing apparatus with stress
US8114670B2 (en) 2005-03-14 2012-02-14 Dfb Technology Holdings, Llc Skin equivalent culture
US9271923B2 (en) 2005-05-26 2016-03-01 Intercytex Limited Tissue repair using allogeneic dermal fibroblasts

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