Abstract
The present invention is a formulation of organic and non-organic components for rapid wound healing and control of infection. The components of the present invention i.e. the formulation are an antimicrobial agent, an antioxidant, a cell proliferation inducing factor and an organic component responsible for matrix formation. Adding these four components in pre-determined proportions, formulations can be made for gels, liquid solution and biodegradable scaffolds which are safe and gives a synergistic effect.
Claims
1. A novel formulation for rapid wound healing and control of infection comprising: Silver Nanoparticles as an antibacterial agent, an antioxidant agent and; Sphingosine-1-phosphate as an inducer of endothelial cell proliferation wherein, the formulation is locally acting topical formulation.
2. A formulation for rapid wound healing and control of infection as claimed in claim 1, wherein said antioxidant agent is selected from Gallic acid, group of Vitamins, Bioflavonoid, Carotenoids, Hydroxycinnamates, Other natural antioxidants like Theaflavin, theaflavin-3-gallate, allicin, piperine, curcumin, Physiological antioxidants, Fungal antioxidants and synthetic antioxidants such as Cinnamic acid derivatives.
3. A formulation of claim 1, wherein concentration of silver nanoparticles is in the range of 0.8 mg/g-1.5 mg/g of gel
4. A formulation of claim 1, wherein the concentration of Gallic acid is in the range of 1 mg/g-5 mg/g of gel
5. A formulation of claim 1, wherein the concentration of Sphingosine-1-phosphate is in the range of 10 μM-1 mM/g of gel
6. A novel formulation for rapid wound healing and control of infection as claimed in claim 1, wherein the size of Silver nanoparticles is in the range of 10 nm to 100 nm
7. A method preparing the formulation as claimed in claim 1, comprising the steps of: i. Adding 10 μM Spingosine-1-Phosphate and 1 mg Gallic Acid to 100 ml of Silver nanoparticles ii. Adding 0.5% of carbopol to the above solution iii. Continuously stirring the above solution on a magnetic stirrer and neutralizing the resultant solution with Triethanolamine
8. The formulation as claimed in claim 1, wherein the formulation can be in the form of a cream, gel, topical solution, patch, ointment, spray or lotion, patch or wipe.
9. Method of preparing the formulation as claimed in claim 8, is carried out at pH 7.0.
Description
BRIEF DESCRIPTION OF DRAWINGS
[0057] FIG. 1 is a graphical representation of antimicrobial activity of the novel formulation against the common infection causing bacteria
[0058] FIG. 2 is a representative of the toxicity of each component in the novel formulation on endothelial cells
[0059] FIG. 3 shows the fibroblast migration when the wound is administered with just lipid formulation, Sphingosine-1-phosphate (1 μM) and Sphingosine-1-phosphate (0.1 μM)
[0060] FIG. 4 shows the quantitative data of fibroblast migration against different concentrations of Sphingosine-1-phossphate and lipid formulation
[0061] FIG. 5 shows antioxidant assay of Gallic acid used in the formulation
[0062] FIG. 6 shows wound healing in natural conditions i.e. control
[0063] FIG. 7 shows wound healing after application of novel formulation
[0064] FIG. 8 shows wound closure percentage in vivo in excision wound model in rats
[0065] FIGS. 9 and 10 represents levels of Lipid peroxidation and Catalase in the wound area.
[0066] FIG. 11 is Photomicrographs of histology from wound site
DETAILED DESCRIPTION OF THE DRAWINGS
[0067] FIG. 1 further explains the antimicrobial activity of each of the components in the novel formulation along with novel formulation, Iodine Tincture and Ethanol against the common infection causing bacteria such as E.coli, E. cloacae, P. mirabilis, S. aureus, S. marcescens, P.aeruginosa, S.pneumoniae by using well plate method which shows larger zone of inhibition when treated with novel formulation as compared to single component such as to silver nano-particles, Gallic acid, sphingosine-1-phosphate and collagen hydrolysates
[0068] FIG. 2 further explains the toxicity of Silver nanoparticles on Human cell lines (Human umbilical cord endothelial cells) after adding individual components such as silver nanoparticles, Gallic acid and sphingosine-1-phoshpate and the novel formulation
[0069] FIG. 3 explains fibroblast migration in the wound healing process after regular intervals of 24 hours to 36 hours after the onset of wound. When the wound is treated with the novel formulation the gap fills fast i.e. the fibroblast migration happens fast
[0070] FIG. 4 explains the quantitative data represented in a graph showing the difference between the novel formulation and control was significant when S1P is added at 1 μM concentration which is responsible for fibroblast migration and also for proliferation
[0071] FIG. 5 further explains the release of Gallic acid as an indicator of stability of the gel over time. The release pattern of Gallic acid is not changing with time over 3 weeks. The Gallic acid release over the period of 300 min is increasing indicating a sustained release from the gel
[0072] FIG. 6 represents the control rat wound healing, excision wound was formed and representative images were taken on 0, 6, 12 and 18 days. Wound healing is observed, however on the 16.sup.th day a little scar observed.
[0073] FIG. 7 represents wound closure in the test rat
[0074] FIG. 8 Current formulation showed a significant increase in wound healing (**p<0.01) as compared to control. The wound healing results for standard gel Plermin used were comparable with the current formulation
[0075] FIG. 9 shows the levels of LPO in rats treated with current formulation was lesser than the rats treated with standard gel. This justifies that the current formulation is effective against the oxidative stress generated during the wound healing procedure.
[0076] FIG. 10 show that the levels of catalase were increased significantly for current formulation treated groups
[0077] FIG. 11 shows that the inflammatory cells neutrophils and macrophages were more in diseased control as compared with the standard control and current formulation. The presence of inflammatory cells confirms prolong inflammation in control which delays wound healing. The presence of these cells shows an early stage of wound healing in diseased control rats while in the rats treated with the current formulation, inflammation has reduced indicating the progress of wound healing. Granulation tissue, proliferation of fibroblast cells and Collagen deposition is more in current formulation than in control and standard gel used (Panel 1 horizontal). The Collagen deposition is more compact in the group of rats treated with current formulation. In the control group the deposition is in patches (Panel 2 horizontal). Further confirmation of collagen deposition is done by using Masson's trichrome stain which stains specifically the collagen. In this staining it is found that rats treated with the current formulation shows good deposition of Collagen as compared with Standard and diseased control (Panel 3 horizontal). This staining confirms the histology results. Immunohistochemistry of proliferation marker Ki67 is done to check the cell proliferation at the wound site. It is observed to be more in animals treated with the current formulation as compared to standard and Diseased control, highlighting the proliferating activity of the current formulation (Panel 4 horizontal).
Abbreviations Used:
[0078] S1P: Sphingosine-1-Phosphate [0079] LPO: Lipid Peroxidase [0080] ROS: Reactive oxygen species
Advantages:
[0081] The novel formulation helps in rapid wound healing even at lower dosage [0082] The novel formulation of present invention eliminates the repeated usage of antibiotics [0083] There is no need of skilled personnel for treatment. [0084] The novel formulation is cost effective and affordable. [0085] There is no adverse reaction of the current formulation when applied topically. [0086] The novel formulation helps the wound to heal holistically faster by addressing bacterial infection, ROS at wound site and angiogenesis
