Effects Of Using

Since many centuries, Ayurvedic medicines have been used by people due to its ability to enhance immunity and prevent diseases.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay  A number of plant, animal, and mineral products are mentioned in Ayurvedic classics for the treatment of different stages of the wound. At the moment, scientific research on medicinal plants is performed most intensely in research institutes, universities, and pharmaceutical laboratories as well as in the clinics of various developed countries. Methods: Major Ayurvedic classics were searched for such useful remedies for wound care which are effective, easily available, and easy to prepare and use them without any side effects. Results: A total of 71 such remedies were found in the Ayurvedic treaties and listed in the article. Conclusion: The article may help the practitioner serving in the remote areas where these plants available abundantly.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

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Abstract
The skin is among the largest and one of the most important organs in the human body. It represents the first line of defence of the body; provides protection from mechanical impacts of the environment, limits the influence of variations in the temperature, prevents entrance of chemicals and microorganisms and restricts radiation effect. Skin damage affects all skin functions; therefore, wounds can compromise patient’s well-being, self-image, working capacity and independence. Due to all mentioned, a good wound management is necessary not only for the individual but also for the community. Herbal medicines have been used to accelerate wound healing since ancient times. Recently, scientists have been able to employ scientific methods to prove efficacy of many of these herbs and to get a better understanding of mechanisms of their actions. The popularity of herbal medicines may be explained by the perception that herbs cause minimal adverse effects. Preparations from traditional medicinal plants in wound management involve disinfection, debridement and the provision of suitable environment for natural healing process. In this chapter, the field of wound healing is briefly introduced. Further, the crucial information regarding plants, which are effectively used as wound healing agents in traditional medicine are gathered.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Keywords
wound physiologywound healingherbsantimicrobial effectanti-inflammatory effectanalgesic effect
Chapter and author infoShow +
1. Introduction
The human body consists of several organs, of which the skin is the largest. The human skin plays an important role in the bodies defensive processes, since it represents the first line of defence [1]. Two other important roles of the skin that also contribute to the defensive mechanisms are regulation and sensation. All mentioned provide a crucial set of functions such as enable protection from mechanical impacts and pressure, restrict the influence of temperature changes, lower the potential impact of microorganisms, limit radiation effects and diminish the entrance of different chemicals. Other important skin functions include the regulation of body temperature (e.g. through sweat glands and hair), control over the peripheral circulation and fluid balance, and in the synthesis of vitamin D. Through its extensive nerve cell network, it enables detection and relaying of changes in the environment (e.g. heat/cold, touch and pain). Damage to these networks is called a neuropathy and impairs the sensation of the mentioned functions in the affected areas. The preservation of skin integrity is due to all mentioned functions crucial for maintaining a healthy body [2].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

A wound is trauma-induced defect of the human skin, involving a multitude of endogenous biochemical events and cellular reactions of the immune system [3]. Wounds can compromise patient’s well-being, self-image, working capacity and independence. Effective wound management is therefore necessary not only for the individual patient, but has an important impact also on the community [4].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2. Mechanism of wound healing
Wound healing is an extremely complex and dynamic process, which includes replacing of devitalized and missing cellular structures and tissue layers. It reflects in a set of biochemical events that integrate into an organized cascade of processes to repair the damaged tissue [5]. Immediately after injury, damaged vessels leak fluid, to which the body responds with haemostasis. Platelets start to aggregate in the wound bed and secrete multiple growth factors that contribute to an effective clot formation to hinder further loss of fluids from the defected area [6]. Simultaneously with the launch of haemostatic mechanisms, the inflammatory phase is induced as well [7]. It is characterized by local vasodilatation, platelet aggregation and phagocytosis, which together with the release of several cytokines, contribute to local inflammation of the wound site. Multiple chemokines, released by platelets, stimulate the immune and other cells (e.g. keratinocytes) to release growth factors and cytokines to regulate various signalling cascades that govern the inflammation and healing in general [8]. Macrophages and other immune cells are stimulated and they migrate towards the wound to dispose cell debris and fight invading bacteria during the wound healing. Angiogenesis occurs at this phase and new blood vessels transport essential nutrients to the wound bed [6]. Effects Of Using Ayurvedic Medication To Heal The Wounds Essay  The next phase in wound healing is the proliferative phase, which is characterized by granulation, wound contraction and epithelialisation. During granulation, fibroblasts form a bed of collagen, followed by the production of new capillaries [7]. During wound contraction, myofibroblasts decrease the size of the wound by gripping the wound edges and pulling them into the wound interior mechanisms that resemble that of smooth muscle cells. After completion of respective processes, unneeded cells undergo apoptosis (controlled cell death) [9]. Epithelialisation is initiated by keratinocytes proliferating and migrating across the wound site [8]. Fibroblasts are activated and differentiated into myofibroblasts that (either indirectly by production of cytokines or directly) regulate other cells to grow and form new epithelial tissue over the wound site. The final wound healing phase is the remodelling phase, which is governed by the rearrangement of the newly formed extracellular matrix (ECM) using increasing amounts of type I collagen. The fibres of collagen rearrange their structures with increasing interfibrillar binding and diameter [10]. The aim of wound treatment can be therefore described as a therapy to either shorten the time required for healing or to minimize the undesired consequences, for example extensive scarring [11]. A general overview of the wound healing Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Figure 1.
Schematic depiction of distinct phases during wound healing.
3. Wound healing management
The complex course of the wound healing with the various physiological events that occur simultaneously, as well as consecutively, is vulnerable to possible external interferences (e.g. infections) on one side, as well enables modulation, and hence improvement of the healing performance, through active treatment solutions (e.g. multifunctional wound dressings) [12, 13]. Among the most desired activities are the ones providing anti-inflammatory, antimicrobial, analgesic and antioxidant activities, regardless of the exact underlying mechanism of action [14].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Shortly after the injury, it is during the acute inflammatory response that different cytokines are formed. These are crucial for orchestration of the specific tissue growth, its repair, and hence regeneration [15]. Nevertheless, if this inflammation step persists, it can negatively affect the wound process, namely it leads to vicious cycle of ongoing inflammation, preventing the wound to reach the remodelling phase. If this happens, delays in wound closure occur, which are often accompanied with the increased sensation of pain in the wounded area and its surroundings that can additionally hinder the healing process [16]. Based on these findings about the wound healing physiology, a lot of research has focused on the development of therapeutic approaches that would provide an anti-inflammatory and pain relieving activity to wound dressings [17].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

An important complication related to wound treatment and healing is infection. Infections are known to significantly increase the treatment costs of wound care [18], which are also the reason that different strategies are being developed for their prevention [19]. Due to the impact of primary and secondary infections on the wound healing, which increase local inflammation, and hence lead to significant tissue destruction, prevention of their occurrence remains one of the main targets of wound dressing development [20]. An ideal medicine for the prevention of wound infection should therefore have antimicrobial activities, while also stimulating the body’s natural immune system without damaging the surrounding healthy tissue [21].

Most wounds induce some level of pain sensation. Pain relates to patient’s discomfort, release of stress hormones and often reduces the patients’ overall quality of life. Hindered mobility and psychological issues connected with pain-induced stress lead to a less effective wound healing. According to McGuire et al. [22], chronic pain lowers the patients’ capability of healing, thereby prolonging the overall recovery process [23]. Suitable and effective pain management can lead to an earlier release from the hospital, stress reduction and a general better reintegration into the community. All mentioned lead to facilitation of wound healing, while at the same time minimizing the risk for development of chronic pain, and finally in lowered treatment costs [24].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Part of the inflammation phase of wound healing causes also a coordinated influx of neutrophils to the wound site. One of the actions of neutrophils is also the activation of the so-called ‘respiratory burst’, which leads to productions of free radicals [25]. These produce oxidative stress that results in lipid peroxidation, DNA damage, and enzyme inactivation (e.g. free-radical scavenger enzymes and others), even those whose main activity is to limit the effects of reactive oxygen species (ROS). Considering the above mentioned, it is clear that antioxidants may be of therapeutic use in several diseases that are connected with potential pathologic actions of oxidants, including the wound healing [26].

Apart from the above-mentioned wound healing aiding activities, others are also reported in literature, e.g. the astringent activity, stimulated epithelisation and effective hydration of the wound site [27]. The most important properties of plant-derived medicines that are beneficial for the wound healing process are depicted in Figure 2 together with some examples of plants that were already proven for the mentioned use.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Aging is a universal process that began with the origination of life many years ago. Skin being the largest organ of the body deserves special attention while considering the aging process. Skin aging is due to the conjunction of intrinsic factors (chronological aging) and extrinsic factors (fundamentally photo aging). In elderly people, cells are less likely to proliferate, they have shorter life spans, are less responsive to cytokines and epithelialization of epithelial skin tissue is longer. Proteases associated with intrinsic aging (metalloproteases) appear increased and less challenged which may predispose to tissue breakdown in the elderly people [1–3]. Thus, to assess integrity of the skin structure, collagenation (collagen synthesis), epithelialization, connective tissue (matrix) deposition and tissue cell regeneration, wound healing animal models can be used [4, 5]. Several studies have been reported to evaluate wound healing activity using different models [6–8]. We have studied the effect of selected topical application forms on tissue regeneration ability and functional status of skin in wound healing models by measuring wound contraction, collagen estimation and breaking strength of the skin.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Ayurveda remains one of the most ancient and yet alive tradition practiced widely in India, Sri Lanka and other countries that have a sound philosophical and experiential basis [9]. Atharvaveda (around 1200 BC), Charak Samhita [10] and Sushrut Samhita (1000–500 BC) are main classics that give detailed descriptions of over 700 herbs. A scholarly description of the legacy of Charaka and Sushruta in contemporary idiom, best attempted with a commentary from modern medicine and science viewpoint, gives some glimpses of ancient wisdom [11]. Ayurveda and traditional Chinese medical system share many common approaches and have a long history of practice [12]. Ayurveda has several formulations for management of aging and related conditions. Ayurvedic literature describes more than 200 herbs, minerals and fats for skin care. We selected cow ghee (butter fat), flax seed oil, Amalaki fruit (Phyllanthus emblica), Shorea robusta resin and Yashada bhasma (zinc complex) for this study. Traditionally prepared cow ghee has two types of properties Samshodhana (cleansing or detoxifying) and Samshana (palliative). Such ghee alone or in combination with honey [13] is considered to be extremely useful for treating wounds, inflammatory swellings and blisters for promotion of quick healing. Ghee is used in many Ayurvedic traditional preparations and also as an ointment base. Flax seed oil is believed to bring mental and physical endurance by fighting fatigue and controlling aging process. It has properties like Madhura (balances the skin pH), Picchaila (lubricous), Balya (improves tensile strength or elasticity of the skin), Grahi (improves moisture holding capacity of skin), Tvagdoshahrit (removes skin blemishes), Vranahrit (wound healing) and useful in Vata disorders including dryness, undernourishment, lack of luster/glow [14]. Both cow ghee and flax seed oil are rich sources of essential fatty acids (EFAs), which regulate prostaglandin synthesis and hence induce wound healing. Effects Of Using Ayurvedic Medication To Heal The Wounds Essay Deficiency of EFA results in phrynoderma or toad skin, horny eruptions on the limbs, poor wound healing, etc. Omega-3 and Omega-6 EFAs are thus important for the maintenance of normal epidermal structure [15–18]. Phyllanthus emblica or Amalaki fruit is a rich source of vitamin C, which is a potent antioxidant [19]. It is foremost amongst the anti-aging drugs (vayasthaprana) or best amongst the rejuvenating herbs; it has properties like Rasayana (adaptogenic), ajara (usefulness in pre-mature aging), ayushprada (prolongs cell life), sandhana karaka (improves cell migration and cell binding) and Kantikara (improves complexion). Juice of the fresh fruit and ghee mixed together is a good restorative tonic. Amalaki helps in fighting many obstinate skin diseases [20]. Shorea robusta resin with Beeswax is used as a base for ointment of herbal extracts used in the healing of foot cracks, psoriasis and other chronic skin diseases. It is useful in wounds, ulcers, neuralgia, burns, pruritus and various skin problems. A combination of cow ghee and Shorea robusta resin is applied in rectal prolapse, haemorrhoids to control burning sensation and excess secretion. An external application made of herbal powders and oleoresin with cow ghee is advocated in blisters to control pain and swelling. Due to excellent healing property, a combination of selected herbal ingredients along with cow ghee and oleoresin is applied on burns and other injuries. It also has anti-inflammatory, antiseptic and anti-microbial properties useful in treatment of skin diseases both infectious and metabolic [21]. Yashada bhasma is a traditional herbomineral preparation mainly comprising of zinc which plays an important role in the normal functioning of skin as it influences the synthesis of collagen. Zinc would benefit the system undergoing rapid cellular division such as wound healing. Studies indicate the epithelization of burns may be improved by zinc treatment. Zinc markedly increases the stability of bio-membranes in general. Ayurvedic literatures describe the activities of Yashada bhasma as Krimighna (antimicrobial), Kantikara (improves complexion), Rasayana (rejuvenator) and Grahi (improves moisture holding capacity of skin). It also improves the binding power of the cells of skin soft tissues, improves cell migration and cell regeneration and hastens wound healing [22–24].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2. Materials and Methods
2.1. Test Material
Cow ghee was obtained from The Nilgiri Dairy Farm Ltd, Bangalore, India; flax seed oil was obtained from Arpitha Aromatics, Bangalore, India; Amalaki fruit extract (spray-dried juice) was obtained from Venkatesh Food Industries, Chhindwara, India; Shorea robusta resin was obtained from M/S Aditya Solvent and Chemicals, Bangalore, India; Yashada bhasma was obtained from M/S Uma Ayurvedics Pvt. Ltd, Kasganj, India. All the samples were identified and confirmed by the Pharmacognosy Department of the Research and Development Center of The Himalaya Drug Company, Bangalore, India. Further identification and authentication of test materials was done by routine or pharmacopoeial methods [25–30] based on parameters such as solubility, pH, loss on drying, ash content, refractive index, specific gravity, saponification value, acid value, iodine value, peroxide value, moisture content, heavy metal analysis, microbial load, zinc content and vitamin C content.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.2. Chemicals
Alpha linolenic acid (ALA) reference standard was purchased from Sigma-Aldrich Inc., USA. All other chemicals and reagents used were analytical grade, unless otherwise specified.

2.3. Formulation of Topical Application Forms
We prepared five variants of the anti-aging topical application form comprising of the study materials in water and in oil emulsion cream. All the variants comprised of the base formulation consisting of Shorea robusta resin and flax seed oil in 1 : 4 proportion. The formula details of the cream variants are given in Table 1. Pharmaceutical grade de-mineralized water and permitted preservative system (methyl paraben+propyl paraben) were used in all test preparations. The stability testing conditions and parameters for the creams were generally based on ICH stability guidelines, WHO stability guidelines and Bureau of Indian Standards. The studies were done in two parts: quick assessment stability studies (from 48 h to 1 month) and accelerated stability studies (6 months). The quick assessment stability studies comprised of thermal stability testing (48 h at 45 ± 1°C/70% RH), freeze thaw testing (five cycles at –5 ± 1°C to 30 ± 1°C) and low temperature testing (1 month at 5°C). The physical parameters including phase separation and color change were observed. The samples were then subjected to accelerated stability conditions (40°C/75% RH) for 6 months and the physicochemical parameters included: pH, viscosity, % total fatty matter content, % water content and % residue. Microbiological stability studies included determination of TVC (total viable count), TFC (total fungal count) and PET (preservative efficacy test or challenge test) [31–34]. Percentage of ALA fatty acid content was also determined in accelerated stability studies.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

NOW

S. No Cream variant code Main ingredients
1 NATAF 001 Shorea robusta resin extract in flax seed oil
2 NATAF 002 Shorea robusta resin extract in flax seed oil + Amalaki fruit extract
3 NATAF 003 Shorea robusta resin extract in flax seed oil + cow ghee
4 NATAF 004 Shorea robusta resin extract in flax seed oil + Yashada bhasma
5 NATAF 005 Shorea robusta resin extract in flax seed oil + Amalaki fruit extract + cow ghee + Yashada bhasma
Table 1

Composition of five variants of anti-aging creams.
2.4. Determination of ALA Content by Gas Chromatography
The ALA content of flax seed oil, cow ghee, Shorea robusta resin extract in flax seed oil and the anti-aging topical application forms was estimated using gas chromatography pharmacopoeial method [26]. The Shimadzu Gas Chromatograph, GC-14 B comprising of flame ionization detector (FID), GC Column—BP × 70 × capillary (SGE) and with s/w—CLASS-GC 10 (version 2.0) was used.

2.5. Chromatographic Conditions
Temperature conditions—Oven temperature—100°C, Injector temperature—130°C, Detector temperature—240°C. Initial 100–160°C at 8°C for 2 min, initial 160–190°C at 10°C for 3 min, initial 190–230°C at 18°C for 6.2 min, total run time—25 min. Gas flow conditions—carrier nitrogen—1 ml/min. Hydrogen—30 ml/min, oxygen—30 ml/min, split ratio—1 : 9.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.6. Standard and Sample Preparation
Reference standard (ALA) 25–100 mg was weighed accurately in 125 mL round-bottomed flask. To this was added 10 mL of pure methanol (HPLC grade) and two to three drops of conc. H2SO4, and the solution refluxed for 3 h at 70°C. After refluxing the contents were cooled to room temperature. It was then extracted with 25 mL of petroleum ether twice. The petroleum ether extracts were collected, combined and filtered through Whatman filter paper (Grade-1, Cat. No 1001917) using anhydrous sodium sulphate. The petroleum ether was evaporated completely at 60°C and the residue dissolved in 5 mL of petroleum ether and made up to 10 mL in volumetric flask and injected into GC. For sample preparation, 2.0–2.5 g of the test sample was weighed accurately, and to this was added 20 mL of 0.5 N methanolic NaOH in a round-bottomed flask and refluxed for 30 min at 70°C using cooling condenser. After refluxing the contents were cooled to room temperature and to it was added 20 mL of water, 1.0 mL of conc. HCl and shaken well. The contents were transferred into separating funnel of capacity 250 mL and extracted with 50 mL of petroleum ether thrice. The petroleum ether extracts were collected, combined and filtered through Whatman filter paper using anhydrous sodium sulphate. The petroleum ether was evaporated completely at 60°C. The residue was taken in round-bottomed flask and to this was added 25 mL of methanol (HPLC grade), four to five drops of sulphuric acid and refluxed for 3 h .Then the method as described for standard preparation was followed. One to two microliters of the sample were injected in GC and the respective components were calculated as:Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.7. Evaluation of Wound Healing Activity
For evaluation of wound healing activity of NATAF creams, excision and incision wound models were used [35, 36].

2.8. Animals
Laboratory-bred (Department of Pharmacology of Research and Development Centre of Himalaya Drug Co., Bangalore, India) rats of Swiss Wistar strain weighing 275–325 g were used for the study. The rats were housed in polypropylene cage and maintained in standard laboratory conditions of temperature (22 ± 2°C) and light–dark cycle of 12 : 12 h. The animals were fed with synthetic pellet diet from Tetragon Chemie Pvt. Ltd, Bangalore, India and water ad libitum during the experiment. The Institutional Animal Ethics Committee (Reg. No 26/1999/CPCSEA) permitted the study for wound healing.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.9. Group Classification
Animals were randomized into six groups of six animals each. The animals of Group 01 were not treated with any cream (to serve as the control group), the animals of Groups 02, 03, 04, 05 and 06 were treated with topical applications of the following creams—NATAF-001, NATAF-002, NATAF-003, NATAF-004, NATAF-005, respectively once daily on the wound area. For both excision wound model and incision wound model the animal groups were classified and treated in the same manner.

2.10. Excision Wound Model
The animals were weighed individually, anaesthetized with pentobarbitone sodium (35 mg/kg, intraperitoneal). The rats were inflicted with excision wounds as described by Morton and Malone [37]. The skin of the dorsolateral flank area was shaved with an electrical clipper. After wound area preparation with 70% alcohol, the skin from the predetermined shaved area was excised to its full thickness to obtain a wound area of about 500 mm. Excision wounds were created on the dorsal thoracic region 1.5 cm from the vertebral column on either side. Hemostasis was achieved by blotting the wound with a cotton swab soaked in normal saline. The respective creams were topically applied on the wound area of the animals of respective groups once a day till complete epithelization; starting from the day of operation. Percentage wound contraction and collagen estimation parameters were studied.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.11. Percentage Wound Contraction
Wound healing is a complex process that results in the contraction and closure of the wound and restoration of the functional barrier. Contractions, which contribute to wound closure, were studied on alternate days from Day 1 to Day 9, that is, starting from the day of operation till the day of complete epithelization by tracing the raw wound on a transparent sheet. The wound area was measured by retracing the wound by the UTHSCSA software Image Tool (Version 3.0.100.0). Falling of the eschar (dead tissue remnants) without any residual raw wound was considered as end point of complete epithelization. Percentage wound contraction was calculated as:Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.12. Collagen Estimation (Hydroxyproline Content)
Wound tissues were analyzed for hydroxyproline content, which is basic constituent of collagen. The collagen composed of amino acid (hydroxyproline) is the major component of extra-cellular tissue, which gives strength and support. Breakdown of collagen liberates free hydroxyproline and its peptides. Measurement of hydroxyproline hence can be used as a biochemical marker for tissue collagen and an index for collagen turnover [38]. For preparation of protein hydrolysate, 50 mg of tissue sample in 1.0 mL of 6.0 N HCl was weighed and sealed in screw-capped glass tube. The tubes were autoclaved at 15 1.056 kilograms per square centimetre for 3 h. The hydrolysate was neutralized to pH 7.0 and brought to the appropriate volume (filtered if necessary). Test tubes marked as sample, standard and blank were taken. One milliliter of test sample was added to test tubes marked as sample, 1.0 mL of DM water to test tubes marked as blank and 1.0 mL standard solutions to test tubes marked as standard. One milliliter of 0.01 M copper sulphate solution was added to all the test tubes followed by the addition of 1.0 mL of 2.5 N sodium hydroxide and 1.0 mL of 6% hydrogen peroxide. The solutions were occasionally mixed for 5 min and then kept for 5 min in a water bath at 80°C. Tubes were chilled in ice-cold water bath and 4.0 mL of 3.0 N sulphuric acid was added with agitation. Two milliliters of p-(dimethylamino)benzaldehyde was then added and heated in water bath at temperature 70°C for 15 min. The absorbance was measured at 540 nm using Synergy HT Multi-Detection Microplate Reader (MDMR). The concentration of the sample was calculated as:Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.13. Incision Wound Model
The animals were weighed individually, anaesthetized with pentobarbitone sodium (35 mg/kg, intraperitoneal) and the skin of the dorsolateral flank area was shaved with an electrical clipper. After wound area preparation with 70% alcohol, two para vertebral long incisions were made through the skin at a distance of about 1.5 cm from the midline on either side. Each incision made was 5 cm long and the parted skin was stitched with interrupted sutures using black braided silk surgical suture (size 3X0) and a curved needle (No. 11) at 1.0 cm interval. The respective creams were topically applied on the wound area of the animals of respective groups once a day till 7th day starting from the day of operation. The sutures were removed on 7th day and the skin breaking strength of the healed wound was measured on 8th day.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.14. Breaking Strength
One of the most crucial phases in dermal wound healing is the progressive increase in biomechanical strength of the tissue; the mechanical properties of the skin are mainly attributed to the function of the dermis in relation to the structure of collagen and elastic fiber networks. Breaking strength of the healed wound is measured as the minimum force required to break the incision apart. Skin breaking strength gives an indication of the tensile strength of wound tissues and represents the degree of wound healing [39].Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Tensile strength has commonly been associated with the organization, content and physical properties of the collagen fibril network. Tensile strength is the resistance to breaking under tension; it indicates how much the repaired tissue resists breaking under tension and may indicate in part the quality of the repaired tissue. After removal of skin sutures on postoperative Day 7, gradually increasing weight was applied to one side of the wound while the other side was fixed. The weight that completely separated the wound from the incision line is considered to be the breaking strength. The sutures were removed on the 7th day after wounding and the breaking strength was measured on the 8th day. The mean breaking strength on the two para vertebral incisions on both sides of the animals were taken as the measures of the breaking strength of the wound of the individual animal.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

2.15. Statistical Analysis
All the values were expressed as mean ± SEM. The values were analyzed using one-way analysis of variance (ANOVA) followed by Dunnett’s post hoc multiple comparison test to establish statistical significance. The analysis was performed using Graph Pad Prism software (Version 4.0).

3. Results
The physicochemical analysis results for the study materials are tabulated in Table 2. The quick assessment stability studies results for the cream variants showed no phase separation or color change for any of the creams and hence were subjected to accelerated stability conditions (40°C/75% RH) for 6 months. The pH of all the variants was stable in the range of 5.0–6.5, which is nearer to the normal physiological pH to ensure better acceptability on application to the skin. Viscosity of all the variants showed a general trend in rise to about 200000–300000 cps over a period of 6 months which is acceptable for the non-Newtonian systems. Other parameters like TFM (total fatty matter), water content, residue, remained constant. Microbiologically, all the variants were stable with TVC and TFC count less than 10 (cfu/g) and all the batches passed the PET analysis which confirms the efficacy of the preservative system.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

Test parameters Cow ghee Flax seed oil Amalaki fruit extract Shorea Robusta resin Yashada bhasma
Description Semi solid, granular in texture, light yellow in color with sweet characteristic odor Liquid, golden yellow in color with characteristic odor Free flowing powder, light greenish brown in color with faint characteristic odor Dried resin pieces, irregularly cylindrical in shape, longitudinally shriveled, brownish to gray in color with faint balsamic odor Fine powder of light yellow color, odorless, tasteless
Solubility Soluble in chloroform, hexane, Dichloromethane (DCM), toluene, sparingly soluble in methanol, insoluble in water Soluble in chloroform, hexane, DCM, toluene, sparingly soluble in methanol, insoluble in water Very slightly soluble in chloroform, hexane, DCM, soluble in water, methanol Soluble in chloroform, hexane, DCM, methanol, slightly soluble in water Insoluble in chloroform, hexane, DCM, methanol, water, freely soluble in 50% HCl
pH Not applicable Not applicable 3.11 5.80 7.29
Wt/ml 0.905 0.926 Not applicable Not applicable Not applicable
RI 1.4590 1.477 Not applicable Not applicable Not applicable
Loss on drying at 105°C (% by wt) Not applicable Not applicable 7.46 1.25 0.049
Total ash content (% by wt) Not applicable Not applicable 4.77 0.049 99.95
Acid value 0.66 0.83 Not applicable Not applicable Not applicable
Iodine value 5.80 5.89 Not applicable Not applicable Not applicable
Peroxide value 1.49 9.1 Not applicable Not applicable Not applicable
Saponification value 230.66 207.30 Not applicable Not applicable Not applicable
Baudouin test Negative Not applicable Not applicable Not applicable Not applicable
Sieve analysis, % retention on 150 μ% retention on 75 μ Not applicable Not applicable Not applicable Not applicable 0.40, 0.59
Bulk density (gm/ml) After first tap After 50 taps Not applicable Not applicable Not applicable Not applicable 1.38, 2.35
Percentage vitamin C Not applicable Not applicable 7.57 Not applicable  Effects Of Using Ayurvedic Medication To Heal The Wounds Essay
Percentage zinc content Not applicable Not applicable Not applicable Not applicable 79.32
Heavy metals (Hg, Cd, As, Pb) Not applicable Not applicable <0.5 ppm <0.5 ppm <0.5 ppm
Microbial load(cfu/gm) TVC TFC Not applicable Not applicable <100, <10 <10, <10 Not applicable
Table 2

Results of physicochemical analysis.Effects Of Using Ayurvedic Medication To Heal The Wounds Essay
3.1. ALA Content
The GC retention time for ALA was found to be 17.531 min. The percentage ALA content for flax seed oil and cow ghee was found to be 6.42% and 1.23%, respectively, for Shorea robusta extract in flax seed oil it was found to be 5.5–6.0%. For the cream variants, the percentage ALA content remained stable for 6 months even in accelerated stability studies as depicted in Table 3.

Variants Initial 1 month 2 months 3 months 6 months
RT 40°C 75% RH RT 40°C 75% RH RT 40°C 75% RH RT 40°C 75% RH
NATAF 001 1.75 1.70 1.69 1.75 1.73 1.71 1.74 1.73 1.71
NATAF 002 1.78 1.73 1.69 1.67 1.63 1.75 1.69 1.77 1.75
NATAF 003 2.73 2.70 2.67 2.72 2.69 2.70 2.65 2.72 2.70
NATAF 004 1.74 1.73 1.73 1.71 1.70 1.69 1.72 1.70 1.70
NATAF 005 2.24 2.18 2.20 2.24 2.19 2.23 2.25 2.23 2.20
Table 3

Percentage ALA fatty acid for accelerated stability studies in test preparations.
3.2. Excision Wound Model
Wound contraction ability in excision model was measured at different time intervals till complete wound healing took place. Table 4 depicts the effect of topical application of NATAF cream variants on percentage wound contraction in excision wound model. Group 05 (treated with NATAF 004) and Group 06 (treated with NATAF 005) exhibited significant (P < .01) increase in the percentage of wound contraction as compared to the untreated control on Day 7 and Day 9. After complete wound healing the hydroxyproline content which is indicative of the collagen turnover was determined in the treatment groups and control group. The results presented in Figure 1 clearly depicts that in treatment Group 05 (treated with NATAF 004) and Group 06 (treated with NATAF 005) the hydroxyproline content was significantly higher (P < .05) compared to the control Group 01. Effects Of Using Ayurvedic Medication To Heal The Wounds Essay

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