NZ719455B2 - Wound treatment composition - Google Patents

Abstract

composition for use in wound treatment is provided comprising (a) sodium meta-silicate, (b) sodium carbonate, (c) sodium gluconate, (d) potassium aluminium sulfate, and optionally (e) sodium chloride, which composition is especially suitable as a rinsing or soaking agent for cleaning surfaces of a wound, opening, loosening and removing biofilms from wound and other body surfaces, dissolving incrustations or scabs from body surfaces and as a dissolving agent for dressings. The composition has a beneficial and synergistic effect on the traditional treatment of wounds with antibiotics or oxidizing agents in that it makes the latter treatment more effective. wound, opening, loosening and removing biofilms from wound and other body surfaces, dissolving incrustations or scabs from body surfaces and as a dissolving agent for dressings. The composition has a beneficial and synergistic effect on the traditional treatment of wounds with antibiotics or oxidizing agents in that it makes the latter treatment more effective.

Description

Wound treatment composition Field of the invention The present invention relates to a novel wound treatment composition which is ally suitable as a rinsing agent for cleaning surfaces of the body, ving incrustations or scabs from body surfaces and as a dissolving agent for dressings.

Background of the invention it is well known in the art of dermatology and wound treatment that, especially in the case of chronic wounds, healing of wounds is strongly delayed by coatings on the wound. These coatings usually consist of wound exudate residues, of shed or enlarged fibrin coatings, necrotic tissue and cell debris. They offer favourable growth conditions for germs which, when pathogenic, may result in infection of the wound.

The biofilm mode of growth of the infecting organisms is one of the obstacles to the healing of chronic wounds (Wolcott and Rhoads, 2008). By definition, biofilms are microbial populations that are attached to a surface, or to the surfaces of other organ- isms, and encase themselves in hydrated extracellular polymeric substance (EPS), which is also referred to as "slime". The chemical and physical ties of EPS will vary, but it is mainly composed of polysaccharides. EPS is also associated with other olecules such as ns, DNA, lipids, and even humic substances (Nielsen et al., 1996, Tsuneda et al., 2003).

Even in the absence of enic germs, it is extremely important for promoting and accelerating wound healing to remove coatings on the wound and thoroughly clean and disinfect the wound. Adhesions to the wound and resulting pain can be d by timely and thorough removal of wound coatings.

Various treatment agents are known which on the one hand clean a wound and remove wound coatings and on the other hand kill off germs, without acting 3O in an allergizing, sensitizing or tissue-damaging manner.

WO 03/004013 discloses a wound treatment agent that contains, in aqueous solution, polyhexamethylene biguanide ("PHMB") and at least one surfactant which is a glycine derivative, in particular a betaine, and/or a sulfosuccinate and/or an amide based on an unbranched fatty acid.

WO 94/27440 describes anti-infective PHMB agents with a mean molecular weight Mw of 2,900 to 15,000, in particular 3,200 to 5,000, which are used as an injury antiseptic and/or —treatment agent or as an crobial, antiviral and/or anti- parasitic agent, preferably by intravenous administration. Compared with previously known PHMB disinfectants, the PHMB disclosed here has a higher mean molecular weight Mw and exhibits an enhanced microbicidal action with lower toxicity. The antiinfective agent disclosed in WO 40 exhibits reduced tissue damage as compared to other anti-infective agents, but its cleaning effect is low. describes a wound dressing comprising a polymer ate and a composition comprising a) at least one antimicrobial active agent and b) an agent that reduces the cytotoxicity, comprising an oil-in-water on that onally has one or more alkanediols and/or one or more glyceryl ethers.

EP-A-1103264 describes a tissue cell -promoting solution comprising water-containing active oxygen as a prime ingredient which promotes the reconstruction of tissues, a process that corresponds to the last of the four main steps involved in wound healing biochemical processes: "blood vessel reaction", "blood vessel coagulation" , "inflammation" and "reconstruction of tissues", which would otherwise have to rely on the natural healing power of the biobody itself.

EP-A-0601891 describes an antifungal and skin healing promoting agent comprising a lorite, a sulfite, a nitrite, a chlorate, hydrogen peroxide, ozone water, a nitrate, and water. discloses compositions comprising (a) metasilicate, (b) carbonate, (c) gluconate, and (d) sulfate for the treatment of human skin to alleviate the symptoms of cosmetic or ologic skin conditions, including acne, a and wrinkling caused by photodamage or conditions d to aging, hormonal imbalances, hyper-pigmentation, melasma, sis, dandruff, or the like. The compositions are also useful for removing biofilms from contact lenses and for inhibiting the growth of microbials that are correlated to the formation of biofilm in the oral cavity. Embodiments of the compositions of the latter uses are, e.g., dentifrices and mouth washes. The potential use of such compositions in wound treatment on the human body is neither taught nor suggested.

Summary of the invention The technical problem underlying the invention is to provide a woundtreatment composition with a favorable cleaning effect which is suitable for cleaning , dissolving tations and removing dressings and similar applications, which in combination with a common medicinal treatment, such as antibiotics treatment, has a curative effect on wound-healing and which is substantially free of undesirable side effects.

This object is achieved by the ition of the invention.

The invention therefore relates to a ceutical composition for use as a wound-treatment agent which comprises (a) at least one silicate, (b) at least one carbonate, (c) at least one gluconate, and (d) at least one sulfate.

In a particular aspect, the present invention provides use of a pharmaceutical composition comprising: (a) from about 0.1% to about 2.5% of a metasilicate , (b) from about 0.1% to about 2.0% of a carbonate, (c) from about 0.1% to about 1.0% of a gluconate, and (d) from about 0.1% to about 1.0% of ium aluminium sulfate, in the manufacture of a medicament for treating a wound on the human body, wherein the medicament is to be applied to a wound, optionally in conjunction with a curative agent.

The compositions according to the ion may further comprise (e) at least one salt selected from alkali metal and/or alkaline earth metal sulfates, borates, bromides, citrates, acetates and lactates. The salt should not interfere with the ical activity of the composition. When other materials are present, the salt should not degrade those materials or interfere with their properties or biological activity. In other words, the salt should be inert with respect to the other components.

The compositions according to the invention show a remarkable and unexpected effect especially when used in aqueous solution as a rinsing agent for cleaning wounds. It has been surprisingly found that pretreatment of wounds with a composition according to the invention has a beneficial and istic effect on the traditional treatment of wounds with antibiotics or oxidizing agents in that it makes the latter ent more effective. The composition is g on the EPS matrix by opening and loosening the debris, so that antibiotics are able to better penetrate and get in a position to effectively combat the ion. Chronic biofilm infections, such as rditis and osteomelitis, often t indefinitely unless the infected material is removed. The invention however supports and improves the effect of antibiotics.

The compositions according to the invention are therefore especially suitable as g or g agents for cleaning surfaces of a wound, for opening, loosening and removing ms from wound and other body surfaces, for dissolving incrustations or scabs from wound and other body surfaces and as dissolving agents for dressings.

[FOLLOWED BY PAGE 3a] Brief description of the drawings Figure 1 is bar graph rating the effects of sterile and non-sterile Aqua Finesse treatment on different bacteria in biofilms of open wounds compared to controls; Figure 2 is an illustration of the Aqua Finesse that was applied on a S. aureus agar plate to determine its possible al effect; Figure 3 is bar graph illustrating the treatment with 0.5% and 2.0% of Aqua e followed by antibiotic treatment Figure 4 is a bar graph and an image showing the treatment of biofilm; and Figure 5 is an illustration showing the al Laser Scanning copy of biofilms in four ent conditions.

Detailed description of the preferred embodiments Preferably, said at least one silicate is an alkali metal silicate selected from the group consisting of sodium or ium metasilicate and sodium or potassium ilicate, and mixtures thereof, of which sodium metasilicate, in particular in the form of its pentahydrate is most preferred.

Preferably, said at least one carbonate is selected from the group consisting of sodium carbonate, sodium sesquicarbonate, sodium bicarbonate and mixtures thereof, of which sodium carbonate is most preferred.

Preferably, said at least one gluconate is selected from the group consisting of ammonium gluconate, lithium gluconate, sodium gluconate, sodium starch gluconate, potassium gluconate, ammonium acid gluconate, sodium acid gluconate, lithium acid gluconate, potassium acid ate, ammonium D-gluconate, lithium D-gluconate, sodium D-gluconate, potassium D-gluconate, gluconic acid, gluconic D-acid, gluconic L- acid, ammonium onate, lithium L-gluconate, sodium onate, potassium L- gluconate, calcium gluconate, calcium acid gluconate, calcium D-gluconate, calcium L- gluconate and mixtures thereof, of which sodium gluconate and potassium gluconate are most preferred.

Preferably, said at least one sulfate is selected from the group consisting of potassium ium e, sodium sulfate, potassium sulfate, lithium sulfate, [FOLLOWED BY PAGE 4] ammonium sulfate, magnesium sulfate, strontium sulfate, aluminium sulfate and mixtures thereof, of which potassium aluminium sulfate is most preferred.

The at least one salt defined in (e) may be a single salt material or a mixture of two or more salts alone. When the carrier matrix contains a mixture of salts, those salts are preferably present in equal amounts, e.g., a mixture of two salts in a 1:1 ratio.

Specific examples of suitable salts include, but are not limited to, sodium acetate, sodium onate, sodium borate, sodium bromide, sodium carbonate, sodium chloride, sodium e, sodium fluoride, sodium gluconate, sodium sulfate, m chloride, calcium lactate, calcium e, potassium sulfate, assium phosphate, potassium chloride, potassium bromide, potassium fluoride, magnesium chloride, magnesium e and lithium chloride. Preferred salts are the inorganic salts, such as the Group 1 or 2 metal sulfates and chlorides. A particularly preferred salt, because of their low cost, is sodium chloride. Sodium chloride maybe substantially pure or in the form of rock salt, sea salt, or dendrite salt. Of these, Reef Crystals synthetic sea salt (Aquarium Systems, Inc.) is most preferred.

Silicates in the meta-forms of pentahydrates (e.g. Na28i03.5H20) are known in the art of cleaning logy as good builders with high pH, buffering capacity and dirt removal. Meta-silicates are known to prevent Ca itation. Silicates are unique among the ions, the common ionic 3D raster of SiO4 is the tetrahedral.

Carbonates, exemplified by soda (N32003), are known as effective rs with a high pH and ive buffering ty. In very hard water precipitation may occur due to Ca and/or Mg carbonates. Soda performs well in dirt removal. Therefore, it is an active component in l of extracellular Polymeric Substances (xPS) and soft not fully closed scaling.

Gluconates, exemplified by sodium or potassium gluconate, are known sequestrants. ed with additives, such as sodium carbonates and/or meta— silicates, gluconates are active in scale removal from surfaces, including the skin.

Sulfates, exemplified by sodium sulfate, are known for their coagulation action.

In still a further aspect of the invention the compositions additionally comprise an emulsifying agent, a surfactant, a thickening agent, or a mixture thereof.

In a preferred embodiment of the invention the compositions further comprise a physiologically acceptable carrier, preferably ed from liposomes, solutions, in particular s solutions, preferably with demineralized water, creams, emollients, ointments, gels, solid formulations and liquid formulations, or diluent.

In a most preferred embodiment of the invention the compositions are preferably used in aqueous solution at a pH which is preferably in the range of about 7.5 WO 49364 to about 9.

As stated before, the compositions according to the invention are especially suitable as rinsing or soaking agents for cleaning es of a wound, for removing biofilms from wound and other body surfaces, for dissolving tations or scabs from wound and other body surfaces and as dissolving agents for dressings.

Without wanting to be bound to any scientific theory,'the inventor believes that the sing effects of the claimed compositions, in particular in cleaning surfaces of a wound and ng biofilms including glycocaiyx produced by some bacteria, epithelia and other ceils, may be explained as follows.

As biofilm formation starts from the first minute in aqueous environment, the first step is the formation of a conditioning film of organic molecules. After initial attachment, planktonic bacteria adhere irreversibly to the e and proliferate to transform into ial colonies. Subsequently bacteria generate a g of XPS, which is essential for the development of the architecture of any biofilm matrix; it provides a framework into which microbial cells are inserted. Confocal laser scanning microscopy indicates that micro-colonies within a biofilm are three-dimensional structures of mushroom-like bacterial growth with water channels running between them for a constant supply of nutrients. The most part of the biofilm, generally around 97%, ts of water. The microbial cells form only about 2-5 % of the m.

Briefly, according to the present invention, two compounds are selected for their builder activity, a meta-silicate and a ate. Both compounds creep behind the biofilm for loosening the xPS from the skin, and actually from the organic molecular layer on the skin. Together with two specific salts known for their coagulation and ed scale removing capabilities, a giuconate and a sulfate, the biofilm and existing scaling are removed with cleansing water.

Both the meta—silicate and carbonate are anionic in their action consisting of absorption to the XPS ng the g forces to their substrate. Bacteria in the xPS have positively charged cell walls as shown by Mera and Beveridge, J. Bacteriol fig 7:1936—1945 (1993). The original double layer of positive and negative les is loosened from the substrate by the meta-silicate and carbonate as they are negatively charged and creep under the xPS. Both meta-silicate and carbonate show to have synergistic action resulting in better cleaning as should be ed from the individual salts. This is expressed in the zeta potential which is still at the same level at some distance from the substrate surface compared to the situation with the xPS stuck directly in the wound. Zeta potential is defined as the electrical potential at the substrate surface with respect to the bulk liquid = water, i.e. the zeta is higher at the surface and becomes lower when the distance between substrate and biofilm or dirt increases. After addition of a composition ing to the invention on the wound, it will seek the lowest ial energetic level. This basic rule in chemistry and physics is reflected in the situation of loosened and coagulated/encapsulated biofilm ed that the dosed concentrations are at the correct levels. In the end, the negatively charged organic layer or substrate is replaced by a negatively charged layer of the anionic salts composition of the ion.

In a further aspect of the present invention the composition comprises, in weight percent: a) from about 0.1% to about 2.5%, preferably about 0.5% to about 2.0%, of a meta- silicate, preferably sodium meta~silicate; b) from about 0.1% to about 2.0%, preferably from about 0.5% to about 1.5%, of a carbonate, preferably sodium carbonate; c) from about 0.1% to about 1.0%, preferably from about 0.3% to about 0.7%, of a gluconate, preferably sodium gluconate; d) from about 0.1% to about 1.0%, preferably from about 0.3% to about 0.7% of a sulfate, preferably potassium ium sulfate; e) optionally, from about 0.1% to about 1.0%, preferably from about 0.3% to about 0.7% of a salt as defined herein, preferably sodium chloride, more preferably synthetic salt. ably, the total percentage of ingredients in the composition is < 5 wt.%.

A particularly preferred ition according to the invention comprises: - sodium metasilicate pentahydrate (PQ Silicas B.V., Netherlands), 1.83 wt.%; - sodium carbonate (Tata Chemicals Europe, UK), 1.43 wt.%; - sodium gluconate 99% unzlauer S.A., ), 0.5 wt.%; - potassium aluminium sulfate >99% (Kaliumaluin solid, Caidic Belgium N.V.), 0.5 wt.%; -synthetic seasalt, containing 58% NaCI (Reef Crystals synthetic t, Aquarium Systems, inc.), 0.5 wt.%; - in an aqueous solution, adding a suitable acid, for example citric acid or acetic acid or hydrochloric acid to adjust the pH to 7.5 to 9, in ular to about 8.

The concentration of the aqueous composition according to the invention use as a wound-treatment agent may vary between ranges which are known in the art or can be d from commercial wound-treatment agents, but is preferably in a dilution of about 1:200, Le. 50 ml of stock solution in 10 liters of water.

The composition for use as a wound—treatment agent can principally be used in any form of preparation known and suitable for wound treatment, which are ointments, tinctures, sprays, g solutions or a washing or shower gel for wound treatment, as a moisturizing gel or as a moist wound covering, as a dissolving gel for dissolving incrustations or scabs from body surfaces or wounds or for removing dressings and for changing moist dressings, and for ng burns and skin transplants.

Preferably, the compositions according to the invention are used as wound- treatment agent in the form of washing solutions, washing gels or gel-like wound coverings. For example, a wound-rinsing solution consists of 98 percent by weight of water, ably 99.5 percent by weight of water.

When used in the form of a wound gel, a preferred ition contains 1 to percent by weight of glycerine and 0.2 to 5 percent by weight of hydroxyethyl cellulose, especially 5 to 12 percent by weight of glycerine and 0.2 to 3 percent by weight of hydroxyethyl cellulose.

When the composition for use as a wound—treatment agent is used in the form of a wound gel, it contains the additives usually used in medical or cosmetic gels.

For the compositions in accordance with the ion for use as a wound-treatment agent, additives of 1 to 15 percent by weight of glycerine and 0.2 to 5 percent by weight of hydroxyethyl cellulose, especially 5 to 12 percent by weight of glycerine and 0.2 to 3 percent by weight of hydroxyethyl cellulose, have proven to be advantageous for the mixtures as described above. in all compositions in accordance with the present invention for use as wound-treatment agents, the additives and admixtures that are usually used for skin— cleaning, wound-cleaning and anti-infective agents can be present in on to the mentioned ents. Other preservatives, colorants, flavors and fragrances, binding agents, moisturizing or g agents, consistency regulators, soiubilizers or the like can be mentioned, for example which can be added in the usual quantities. it is preferable however to keep the number and concentration of the additives and admixtures as low as le or to avoid the same entirety. it is understood that substances which lead to a reduced effect of the components already present or have a disadvantageous effect on the tissue to be cleaned shall be avoided as far as possible.

Either cleaned water or another s solution as used otherwise in the field of medicine is suitable as a solvent in the compositions according to the invention for use as a treatment agent. Suitable solvents are for example a saline or Ringer's on. A saline solution of 0.4 to 1.2 percent of weight can be used for example. The saline or Ringer's on is used especially preferably in a physiological dilution. In the case of solutions, the invention comprises both the solutions y d and concentrates which need to be diluted priorto use.

The composition in accordance with the invention for use as a wound— treatment agent is especially suitable for treating chronic wounds, as occur for example in diabetics or bedridden patients. Diluted s solutions can be used for rinsing or bathing wounds or for soaking and humidifying wound coverings. Adhering wound dressings could be d easily from the wound by softening with the solution without damaging the wound tissue. When dressings are d it often occurs that lncrustations already formed are torn open again in an undesirable manner during the removal of the dressing, thus leading to delayed g of the wound. The composition in ance with the invention for use as a wound—treatment agent provides a remedy for this purpose too. When the composition in accordance with the invention for use as a wound-treatment agent is applied to the dressing, it penetrates and detaches the same from the incrustations on the wound, so that the dressing can be removed without damaging any incrustations already formed. At the same time, it is prevented that bacteria or the like are implanted in the wound.

The composition in accordance with the invention for use as a wound- treatment agent in gel form can be used as a moisturizing gel or moisture wound covering. Wound coatings can thus be detached, the wound can be cleaned. One application is the removal of scabs in patients“ noses who are artificially respirated. The gel is also especially suitable for treating burns and skin transplants in order to keep the wound humid and smooth and to prevent forming breeding ground for bacteria and other germs. Application can be made directly on the wound or on a wound dressing.

The gel layer ly prevents recontamination of disinfected wounds by germs introduced from the outside.

The main field of application of the composition according to the invention for use as a wound-treatment agent is, as mentioned before, the cleaning of wounds, wound conditioning and wound remediation. Main focus must be laid on the removal of wound coatings which delay the healing of wounds and form a breeding ground for new ment of pathogenic germs on non-infected wounds.

Traditionally focus on planktonic cells in wound g is outdated science.

The focus of the composition according to the ion is based on wound bioburden.

Naturally occurring bacteria attached to surfaces rarely behave like planktonic bacteria. Up to 80% of human ious diseases are biofilm related (NlH). The remaining 20% are microbial cells that reside within a microbial community d within the EPS matrix.

Antibiotics fail to eradicate bioflms due to poor penetration, metabolic inhibition and the immune response to antimicrobials.

The ition according to the invention provides for the first time a composition for use as a treatment agent which es a favorable cleaning effect as a pretreatment of wounds with the traditional treatment of wounds with antibiotics or oxidizing agents, eg. a peroxide ointment, in that it makes the latter treatment more effective, t leading to irritations or even destruction of the tissue.

WO 49364 Wounds are reliably cleaned, odor ion is suppressed, and wound healing is promoted.

The invention will now be explained in further detail with reference to the following examples.

Example 1 In this example it is trated that a preferred composition according to the invention, hereinafter named "Aqua Finesse" is effective in the ent of biofilms in vitro and in chronic wounds.

Aqua Finesse had the same composition as the preferred composition sed in the general description: - sodium metasilicate pentahydrate, 1.83 wt.%; - sodium carbonate, 1.43 wt.%; - sodium gluconate (99%), 0.5 wt.%; - potassium aluminium sulfate (>99%), 05 wt.%; - tic seasalt (containing 58% NaCl), 0.5 wt.%; in an aqueous solution, to which citric acid was added to adjust the pH to 8.0.

The following strains of bacteria were tested on treatment with "Aqua Finesse": - lococcus aureus, — Staphylococcus epidermidis - Enterococcus faecalis - Proteus vulgaris - Enterobacter cloacae These bacteria are all to be found in open wounds. Many infections are polymicrobial. The results showed that "Aqua Finesse", both as a sterile and non-sterilized product, performs efficiently on the biofilms. See Figure 1 Pure "Aqua Finesse" was applied on S. aureus agar plate to determine its possible biocidal effect. However, no inhibition zone was found. See Figure 2.

A slime-test was performed with the above five bacteria s and mixtures thereof, and with Pseudomonas aeruginosa at two different doses of "Aqua e" to assess if it dissolved the “slime” layer of the biofilm. All these tests proved that "Aqua Finesse" had effect in removing the slime layer and making biofilms susceptible to antibiotics.

An MTT assay was performed with the separate bacteria biofilms and a biofilm of the mixture exposed to 3 different doses of "Aqua e" for 5 and minutes. An M'l‘l' assay was also performed to assess the effectiveness of treatment with a combination of "Aqua e" and antibiotics. AftenNards a confocal laser scanner microscope was used. it was demonstrated that antibiotic treatment is more ive after pre-treatment with "AquaFinesse".

It was further sing to note that the test with S. aureus showed a biofilm left, but it became susceptible for antibiotics. This bacteria is normally ant against antibiotics. It was concluded that removing the outer layer of the biofilm may be sufficient for a successful treatment of the bacteria with antibiotics.

An MTT assay showed that the metabolic activity of the bacteria does not change when exposed to "Aqua Finesse" in different concentrations. Performing multiple tests showed that treatment with "Aqua Finesse" lly opens and loosens biofilm slime and resulted in fewer biofilm clumps.

MTT assay analysis showed prophylactic use of "Aqua Finesse" to have no noticeable effect. rmore treatment of biofilms with only "Aqua Finesse" or antibiotics showed no significant improvement, whereas treatment with "Aqua Finesse" followed by otic treatment showed significantly better results. See Figs. 3 and 4.

Confocal laser scanning microscopy showed that the biofilm ess of the mixture of S. aureus and Ps. Aeruginosa treated only with "Aqua Finesse" decreased the least as compared to the control group. The biofilm exposed to antibiotics and the ation of "Aqua Finesse" and antibiotics showed a reduction in thickness. The CLSM confirmed that the combination of "Aqua Finesse" and antibiotics was the most effective against the biofilms by showing more dead bacteria, although it did not tely remove the biofilm. Although some biofilm was still present it was not attached to the surface anymore.

The mixed biofilm of S. aureus and Pseudomonas was used for the Confocal laser scanning microscopy (CLSM). The biofilm was tested in 4 different conditions. The first well was filled with an untreated biofilm of the mixture as control. In the second well antibiotics were added to the biofilm. "Aqua Finesse" was added to the third well. The fourth well was treated with a combination of "Aqua Finesse" and the otics. Figure shows the CLSM of the biofilm in the different conditions. The green dots show the living cells and the red dots show the death cells. The e shows that "Aqua Finesse" in combination with the antibiotic kills many cells.

Example 2 A stock solution of a composition according to the present invention was made as follows. 80 liters of demineralized water aline) were loaded into a vessel, to which the following ingredients were added under uous stirring to prepare a 50% stock solution: - Artificial sea salt (reef Crystals) 200 g - Sodium gluconate 200 g - Potassium aluminium sulfate ("kalialuin) 200 g — Sodium carbonate 573 g — Sodium metasilicate 733 g This solution was made sterile by gamma irradiation and used to fill 2 litre PE bottles, which were then sealed and provided with a batch number.

Example 3 Cytotoxicity of the solution prepared in Example 2, hereinafter also named Solution A, is assessed by three independent methods in four different human fibroblast cell lines. Cell ity is ined using the Alamar Blue cell viability stain, the percentage of surviving cells after exposure is determined by cell count and the ability of cells to survive and recover after re is assessed using a clonal al assay. Cells were exposed to the test solution for 30 minutes and cytotoxicity was determined 24 hr and 48 hr after exposure. All assays are performed in three independent tests.

Exposure of human fibroblasts to the tested Solution A resulted in a sed cell survival in some of the xicity test, indicating cytotoxic properties of the on.

Cytotoxicity was observed in both primary and immortalized human fibroblast cell lines and was most pronounced in the clonal cell survival. Cytotoxicity was also observed for the alized MRC-5 lung lasts (Huschtscha and Holliday, 1983) and VH—25 primary skin fibroblasts (Abrahams et al., 1992) in the Alamar Blue viability stain for the MRC—S cells in the cell count assay.

It was concluded that Solution A can induce cytotoxicity in cultured human fibroblasts, but the cytotoxic potency strongly depends on the cell type.

Example 4 The efficacy of Composition A to ms was experimentally tested in a static system with different concentrations of the composition in sterile and non-sterile form (50%, 25%, 10%, 5%, 1%, 0.5% and 0.1% solutions in sterile MQ) The following biofilm-generating microorganism strains were used: S. aureus ATCC 6538 and MRSA 8-325, and S. epidermis 1457 and 0068.

It was found that both 8. mis strains formed more extensive biofilms than the S. aureus strains. However, for none of the concentrations of the tested compositions a significant reduction in the ion of the amount of biofilm was found, neither for treatment of 10 minutes, nor for the treatment of 30 minutes.

Of the four tested strains, S. aureus MRSA 8-325 formed the least extensive biofims, but also with this strain no icant reduction of the amount of biofiim was observed at any of the concentrations of the tested composition, neitherfor the ent of 10 minutes, nor for the treatment of 30 minutes. However, for S. aureus ATCC 6538 a clear and significant reduction was observed for the treatment of 10 minutes and 30 minutes at the 25%, 10%, 25%, 10% and 5% concentrations in a decreasing order with decreasing trations. No difference in efficiency was found between the sterile and non—sterile solutions. A longer treatment did not result in this case in an increase in the number of effective concentrations.

The dosage which showed clear effects on the specific m of a bacterial strain (S. aureus ATCC 6538) did not result in cytotoxic effects on the human fibroblast cell cultures.

Claims (20)

Claims

1. Use of a pharmaceutical composition comprising: (a) from about 0.1% to about 2.5% of a meta-silicate, 5 (b) from about 0.1% to about 2.0% of a carbonate, (c) from about 0.1% to about 1.0% of a gluconate, and (d) from about 0.1% to about 1.0% of potassium aluminium sulfate, in the manufacture of a medicament for treating a wound on the human body, wherein the medicament is to be applied to a wound, optionally in conjunction with a curative 10 agent.

2. The use according to claim 1, wherein the composition further comprises (e) at least one salt selected from alkali metal and/or alkaline earth metal sulfates, des, borates, bromides, citrates, acetates and lactates.

3. The use ing to claim 2, wherein said at least one salt is sodium chloride.

4. The use according to any one of claims 1-3, wherein the ition is in 20 the form of an aqueous solution having a pH of 7.5 - 9.

5. The use according to any one of claims 1-4, wherein said ilicate is an alkali metal silicate selected from the group consisting of sodium metasilicate or potassium metasilicate, and mixtures thereof.

6. The use according to any one of claims 1-5, n said at least one carbonate is selected from the group consisting of sodium carbonate, sodium sesquicarbonate, sodium bicarbonate, and es thereof. 30

7. The use ing to any one of claims 1-6, wherein said at least one gluconate is selected from the group consisting of ammonium gluconate, lithium gluconate, sodium gluconate, sodium starch gluconate, potassium ate, um acid gluconate, sodium acid gluconate, lithium acid gluconate, potassium acid gluconate, ammonium D-gluconate, lithium D-gluconate, sodium D-gluconate, 35 potassium D-gluconate, gluconic acid, gluconic , gluconic L-acid, ammonium L- gluconate, lithium L-gluconate, sodium L-gluconate, potassium L-gluconate, calcium gluconate, calcium acid gluconate, calcium D-gluconate, calcium L-gluconate, and mixtures thereof.

8. The use according to any one of claims 1-7, wherein said at least one sulphate is selected from the group ting of potassium aluminium sulfate, sodium 5 sulfate, potassium sulfate, lithium sulfate, ammonium sulfate, magnesium sulfate, strontium sulfate, aluminium sulfate, and mixtures thereof.

9. The use according to claim 1 or 2, wherein the composition comprises (in wt. percent) 10 a) from about 0.5% to about 2.0% of the meta-silicate; b) from about 0.5% to about 1.5% of the carbonate; c) from about 0.3% to about 0.7% of the gluconate; d) from about 0.3% to about 0.7% of the sulfate; e) optionally, from about 0.3% to about 0.7% of sodium chloride.

10. The use according to claim 9, wherein the meta-silicate is sodium metasilicate , the carbonate is sodium carbonate, the ate is sodium gluconate, and the sodium chloride is synthetic sea salt. 20

11. The use according to any one of claims 1-10, wherein the composition further ses an emulsifying agent, a surfactant, a thickening agent, or a mixture thereof.

12. The use according to any one of claims 1-11, wherein the composition 25 further comprises a logically acceptable carrier.

13. The use according to claim 12, wherein the physiologically acceptable carrier is ed from liposomes, aqueous solutions, , ents, ointments, gels, solid formulations and liquid formulations, or diluent.

14. The use according to any one of claims 1-12, n the composition is in the form of a liquid, cream, oil, gel, fluid cream, lotion, emulsion or micro-emulsion.

15. The use according to any one of claims 1-12, wherein the medicament is 35 in the form of a wound-rinsing solution, which consists of at least 98 percent by weight of water.

16. The use according to claim 15, wherein the water is ralized water.

17. The use according to any one of claims 1-12, wherein the medicament is in the form of a wound gel, which contains 1 to 15 percent by weight of glycerine and 5 0.2 to 5 percent by weight of hydroxyethyl cellulose.

18. The use according to claim 17, wherein the wound gel contains 5 to 12 percent by weight of glycerine and 0.2 to 3 t by weight of yethyl cellulose. 10

19. The use according to any one of claims 1-12, wherein the medicament is in the form of a rinsing solution or a washing or shower gel for wound treatment, or a moisturizing gel or a moist wound covering, or a dissolving gel for dissolving incrustations or scabs from body surfaces or wounds or for removing dressings or for changing moist dressings, or for treating burns and skin transplants.

20. The use according to any one of claims 1-19, wherein the curative agent ses at least one of an antimicrobial agent and an oxidizing agent. ..