Composition comprising collagen and honey

Abstract

A composition comprising collagen and honey can be produced by addition of an aqueous solution of honey to an aqueous suspension of collagen. The composition is for use in medicine and particularly for use in the treatment of a wound. A method for treatment of a wound comprises administering the composition topically to the skin of a patient.

Claims

1. A free-standing, biodegradable honey and collagen film produced from a dried aqueous suspension, the aqueous suspension comprising 0.1% to 2.0% bovine collagen by weight and 0.05% to 5.0% honey of Unique Manuka Factor (UMF)+16 by weight; wherein the free-standing, biodegradable honey and collagen film comprises substantially bovine collagen, honey of UMF+16, and water suitable for the treatment of a wound.

2. The free-standing, biodegradable honey and collagen film of claim 1 wherein the wound is inflicted by a burn.

3. The free-standing, biodegradable honey and collagen film of claim 1, wherein the composition further comprises one or more additional medicaments for the treatment of a wound.

4. The free-standing, biodegradable honey and collagen film of claim 1, wherein the free-standing, biodegradable honey and collagen film: (a) has a thickness in the range of 5 μm to 2 μm; or (b) is transparent or translucent; or c) has a thickness in the range of 5 μm to 200 μm and is transparent or translucent.

5. An island dressing comprising the free-standing, biodegradable honey and collagen film of claim 1, wherein the composition is surrounded by an additional supporting thin film.

6. The island dressing of claim 5, wherein the additional supporting thin film: (a) is transparent or translucent; or (b) is a polyurethane (PU) film; or (c) is transparent PU film or translucent PU film.

7. The island dressing of claim 5, wherein the additional supporting thin film has adhesive applied to a body facing surface and provides peripheral adhesion to skin of a patient.

8. The island dressing of claim 7, Wherein the adhesive is acrylic based or silicone-based.

9. The island dressing of claim 7 wherein the adhesive is provided with at least one release liner which covers the adhesive on the body facing surface of the film until the film is ready tier use, and wherein the release liner optionally comprises a tab which does not adhere to the adhesive.

10. The free-standing, biodegradable honey and collagen film of claim 1, wherein the aqueous suspension comprises 0.5% to 1.0% bovine collagen by weight and 0,18% to 1.8% honey of UMF+16 by weight.

11. The free-standing, biodegradable honey and collagen film of claim 1, wherein the aqueous suspension comprises 0.5% to 1.0% bovine collagen by weight and 0.45% to 0.9% honey of UMF‥16 by weight.

12. The free-standing, biodegradable honey and collagen film of claim 1, consisting essentially of bovine collagen, honey of UMF+16, and water.

13. The free-standing, biodegradable honey and collagen film of claim 1, consisting essentially of bovine collagen, honey of UMF+16, water, and an additional medicament.

14. The free-standing, biodegradable honey and collagen film of claim 1, consisting of bovine collagen, honey of UMF+16, and water.

15. A method for production of a free-standing, biodegradable honey and collagen film, the method comprising (a) preparing. a bovine collagen suspension, (b) homogenizing the collagen suspension, (c) preparing a honey solution, wherein the honey is of Unique Manuka Factor (UMF)+16, and (d) adding the honey solution to the collagen suspension to provide a collagen and honey suspension, wherein the collagen and honey suspension comprises 0.1% to 2.0% bovine collagen by weight and 0.05% to 5.0% honey by weight; (e) homogenizing or blending the collagen and honey suspension, and (f) drying the collagen and honey suspension to provide a free-standing, biodegradable honey and collagen film comprising substantially bovine collagen, honey of UMF+16, and water suitable for the treatment of a wound.

16. The method of claim 15, further comprising physically cross-linking or chemically cross-linking the free-standing, biodegradable honey and collagen film.

17. The method of claim 15, further comprising administering the free-standing, biodegradable honey and collagen film topically to a wound on the skin of a patient thereby treating the wound.

18. The method of claim 17 wherein the free-standing, biodegradable honey and collagen film is applied to the wound for about 2 to about 7 days and then reapplied depending on the severity of the wound until the wound is healed.

19. The method of claim 15, wherein the collagen and honey suspension. comprises 0.5% to 1.0% bovine collagen by weight and 0.18% to 1.8% honey by weight.

20. The method of claim 15, wherein the collagen and honey suspension. comprises 0.5% to 1.0% bovine collagen by weight and 0.45% to 0.9% honey by weight.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The invention will now be described by way of example with reference to the accompanying drawings, in which:—

(2) FIG. 1 shows a schematic diagram describing stages of wound treatment;

(3) FIG. 2 shows a schematic diagram showing the stages in a method for production of a composition according to the invention; and

(4) FIG. 3 shows photographs of films having compositions according to the invention.

DETAILED DESCRIPTION OF THE INVENTION

(5) It will be appreciated that aspects, embodiments and preferred features of the invention have been described herein in a way that allows the specification to be written in a clear and concise way. However, it will be appreciated that the scope of the invention may include embodiments having combinations of all or some of the features described. Unless circumstances clearly dictate otherwise, aspects, embodiments and preferred features can be variously combined or separated in accordance with the invention. In a preferred embodiment, a composition in accordance with the invention comprises all aspects of the invention.

(6) The word “about” is taken to mean optionally plus or minus 20%, more preferably optionally plus or minus 10%, even more preferably optionally plus or minus 5%, even more preferably optionally plus or minus 2.5%, most preferably optionally plus or minus 1%.

(7) Within the context of this specification the word “comprises” is taken to mean “includes, among other things”. It is not intended to be construed as “consists of only”.

(8) Within the context of this specification, the word “substantially” means preferably at least 90%, more preferably 95%, even more preferably 98%, most preferably 99%.

(9) Within the context of this specification, “treatment” is taken to mean treatment of an existing condition or disease and/or prophylactic treatment in order to prevent incidence of a condition or disease. As such, the methods of the invention can be used for the treatment, prevention, inhibition of progression or delay in the onset of disease. “Treatment of a wound” is taken to mean reduction in the size or severity of the wound or the prevention or reduction of infection to the wound.

(10) The term “wound” is taken to mean an injury in which skin is damaged, torn, cut, or punctured, or where blunt force trauma causes a contusion. It includes a sharp injury which damages the dermis of the skin or a burn which damages the skin.

(11) Without being bound by theory, it is considered that collagen within a wound environment provides an alternative source of collagen (sacrificial substrate) that can be degraded by the high levels of MMPs (matrix metalloproteinases) allowing the endogenous collagen to continue normal wound healing. The collagen also finds roles in cellular signalling, differentiation, migration, angiogenesis and in the induction of growth factors to aid healing.

(12) In addition, it is considered that in embodiments comprising honey, the honey in the composition may act to reduce water content, creating a more hostile environment for potential bacterial growth. Furthermore, the presence of honey leads to a low pH and this may serve to inhibit microbial growth while increasing oxygen diffusion and decreasing protease activity.

(13) Matrix metalloproteinases (MMPs) play an integral part in the wound healing process. However, elevated levels of these MMPs break down healthy tissue and render wounds as non- or slow-healing. Furthermore, acidic pH has been shown to decrease protease activity and this may promoted by application of honey to induce lower wound pH levels during healing. This lower pH has also been shown to increase oxygen diffusion, another influential factor in wound healing, with this increase in oxygen availability also potentially promoting osmotic pressure induced by sugars in honey, which can act to reduce oedema.

(14) As shown in table 1, both collagen alone and honey alone have previously been shown to have benefit in wound healing. In this regard, table 1 shows a summary of material properties and modality of action for collagen, honey and a novel combination of collagen and honey.

(15) As shown in FIG. 1, the stages of wound treatment are as follows: 1—Dressing is placed on and around wound area 2—Collagen degrades and honey presents itself to the wound 3—Protease and collagenases break down collagen and further exposure of honey to the wound occurs 4—Anti-inflammatory and anti-bacterial activity of honey and biochemical action and cues of collagen facilitate wound healing 5—Regenerated epithelial layer formed with no toxic residues or synthetic particulates.

(16) The figure shows a schematic illustrating application of a dressing over a wound bed and gradual breakdown and release of the components into the wound over time.

(17) A: Sterile dressing is placed over the wound site forming a protective barrier. This may be covered and securely held in place by a secondary dressing or may form an island dressing.

(18) B: as the dressing comes into contact with the wound site, the dressing components, namely collagen and honey, over time begin to interact with the wound. The honey may serve to lower pH, increase oxygen diffusion and decrease protease activity. The honey may also serve to create an osmotic pressure which can promote anti-inflammatory action and moist wound healing. The collagen will serve as a sacrificial substrate while also providing cues including those necessary for cellular differentiation, migration and angiogenesis.

(19) C: over time the collagen begins to break down by the action of matrix metalloproteinases; this biodegradation makes the collagen particulates available in deeper wound site and neo-tissue forms to replace the collagen substrate. The honey continues also to serve as an antibacterial component throughout the healing process.

(20) Compositions according to the invention were prepared as follows:

(21) Materials and Methods

(22) Collagen Suspension Preparation

(23) A 0.5 wt % collagen suspension was prepared by hydrating 2.5 g of type I bovine Achilles tendon collagen. in 500 ml of 1 mM hydrochloric acid (HCl) for 24 hours and blending for 5 hours. Once hydrated, the collagen was transferred into a blender container, placed within an ice-filled Pyrex dish and blended initially using a double-blade blender for 2 hours, followed by blending with a homogeniser for a further 3 hours, to produce a white collagen suspension.

(24) Honey Solution Preparation

(25) Honey concentrations of 0.2, 0.1, 0.05, 0.02 and 0.01 g/ml in distilled water were prepared using honey of Unique Manuka Factor (UMF)+16. The highest honey content was 1.0 g/5 ml=0.2 g/ml (to achieve 25 ml, 5 g of honey was used). A syringe was used to transfer 5 g of honey into a 25 ml volumetric flask placed onto a weighing scale (Denver Instrument) and on filling of the volumetric flask with 25 ml of distilled water a magnetic stirrer was used to mix the solution for 1 hour. This yielded a 0.2 g/ml honey solution, which was used to make up solutions having the lower concentrations by sequential dilution.

(26) As shown in FIG. 2, the method steps for production of compositions according to the invention were as follows: (1) Lyophilised collagen may take the form of a powder, fibre, granules or other forms. The collagen may be type I, II, III, IV or other. It may be purchased pre-prepared or may be extracted from cartilage. The collagen may also be of a denatured form such as gelatin. (2) The collagen may be hydrated in hydrochloric acid, acetic acid or any other acidic solution at pH<7. The duration of hydration may vary according to quantity of collagen and contact area between the solute and solvent. This hydration may typically take between 2 hours and 24 hours. The hydrating collagen should be left refrigerated. (3) The hydrated collagen was homogenised as described above, using a blender and/or homogeniser. Blending time depended on collagen weight fraction and volumes. The blending solution was kept cool and this may be achieved using an ice-bath to surround it. (4) Honey which may take the form of a viscous or semi-viscous liquid or powder can be used. The honey may be sourced from the Leptospermum Scoparium bush, indigenous to New Zealand, which has shown to exhibit unique antibacterial and anti-inflammatory properties and to promote wound healing. The honey may also be from different sources and may exhibit varying degrees of bioactivity. (5) A honey solution was made up as described above using distilled water in order to aid accurate transfer of specific honey quantities. (6) The honey solution was homogenised using a blender and or homogeniser and a magnetic stirrer may also be used. Physical agitation, by shaking, may also be used. (7) The honey solution was added to the homogenised collagen suspension using a pipette. The honey solution was transferred drop-wise with continual homogenisation or it was added in bulk and then blended and/or homogenised. (8) The resulting collagen-honey suspension can be homogenised. (9) The resulting collagen-honey suspension was degassed in a vacuum oven, chamber or other facility to remove air bubbles. (10) The collagen and honey suspension was poured using a pipette into a mould, tray or onto paper. (11) The poured suspensions were left to degas (12) The suspensions were dried in an oven or chamber at low temperatures (about 20° C. to about 35° C.) or at higher temperatures (above about 35° C.). The suspension was also lyophilised or freeze-dried to form porous sponges or membranes. (13) The resulting composition comprising collagen and honey in the form of a film was provided (14) Physical or chemical cross-linking may be used to impart strength and/or other desirable properties to the film as required. The cross-linking method may include but is not limited to treatment with glutaraldehyde and/or other chemical cross-linking agents and optionally dehydrothermal treatment.

(27) Results

(28) Compositions according to the invention were prepared as described above. The collagen and honey weight percentages and ratios of six thin films, fabricated under the same conditions are shown in Table 2.

(29) Each film identified in Table 2 was fabricated using 20 ml of collagen and honey suspension and dried in a vacuum oven for four days under a temperature of 30° C. at ambient pressure.

(30) TABLE-US-00002 TABLE 2 Collagen:Honey Ratio Control 1:0 5:1 5:2 1:1 1:2 1:4 1 2 3 4 5 6 Collagen 0.45 0.45 0.45 0.45 0.45 0.45 wt. % (0.25 g) (0.25 g) (0.25 g) (0.25 g) (0.25 g) (0.25 g) (weight in grams/50 ml) Honey wt. % 0   0.09 0.18 0.45 0.9  1.8  (weight in (0.05 g)  (0.1 g) (0.25 g)  (0.5 g)  (1.0 g) grams/50 ml) Description A thin, highly A thin A translucent A translucent A A translucent transparent transparent film with yellow film with yellow translucent and adhesive and and tint and small tint and small film with film with good homogenous homogenous dispersed dispersed yellow tint surface clear film. No near clear agglomeration. agglomeration. and homogeneity. air bubbles. film with little Material exhibits adhesive Material feels surface adhesive behaviour. less brittle irregularity properties and Material is compared and a single more difficult to difficult to with sample 1 air bubble. remove from remove (control) mould. from mould.

(31) Films having various compositions identified in Table 2 were prepared in accordance with the invention and they are shown in FIG. 3. The films were compared and comparative test data shows that: The control scaffold appeared consistent with previously fabricated films under the same conditions The transparency of the composition decreased with increasing honey content There was increased adhesion of films comprising compositions according to the invention with a Teflon mould and associated difficulty in removal of the compositions from the mould with the addition of honey The films with honey were felt to be more robust than the control film

(32) The above described embodiments have been given by way of example only, and the skilled reader will naturally appreciate that many variations could be made thereto without departing from the scope of the claims.