Authored by R Sridhar Skylab*
Abstract
Skin is the most important organ of the human frame. It acts as abarrier and protection to the complete human body. When skin gets injured, the repair process entails removal of the damaged tissue and laying down of a new extracellular matrix (ECM) over which epidermal continuity can be re-established. Burns can cause many of the reasons. It can cause tremendous problems for themselves and their families. Burns are classified according to its depth of severity. Nowadays industries are moving towards the water origin collagen, particularly the fish to create a scaffold for wound treatments. Instead of deriving collagen from fish, the fish skin acts as the scaffold. Tilapia fish is rich in collagen which is used to create a scaffold to treat wounds. This is the first time that the origin the collagen is directly applied on the skin for wound healing. Now a day’s modern medicines are derived from medicinal value herbs. Herbs have its own medicinal values. In this study, tridax procumbens is chosen because of its higher wound healing and reepithelialization property. The herb is extracted and blended with tilapia’s skin to minimize the healing time. FTIR studies are confirmed that the components responsible for wound healing are present in both tilapia skin and extract of tridax procumbens. Structural studies are done by SEM analysis. The breaking point of that skin is found by tensile strength analysis. The biodegradation study revealed that the scaffolds mass reduced to 50% in 20 days. After 35th day the rate of reduction in mass was very less when compared with previous days. MTT study revealed that the coated scaffold has less toxicity when compared to the coated scaffold.
Keywords: Burns; Collagen; Degradation; Scaffold; Tridax procumbens; Tilapia; Wound healing
Introduction
Many advancements are arising to treat the wounds, especially for burn wounds. Wounds that can cause psychological and economic problems in human life. However, the treatments get advanced day by day. The general classification of wounds is shown in bellow.
Now a days industries are moving towards the collagen origin, which are all cost effective to the application of wounds. Collagen type-I are responsible for fibril formation, which leads to faster healing. Wound healing contains three major phases.
Phases of wound healing
• Inflammatory (reactive)
• Proliferative (reparative)
• Maturation (remodeling) (Figure 1)
If the skin gets injured, the body responds easily. The wound healing time is depending on the vascular and cellular response of the injury. The blood vessels are constrict within a few seconds in the injured area to avoid blood loss. Platelets are accumulated in the injured area and hence clots the blood. To expose the sub endothelial collagen surface, the endothelial cells are retracted. In that surface the platelets are attached. Through the adhesive glycoproteins (fibrinogen, fibronectin, thrombospondin and von willebrand factor) the adherence occurs between exposed collagen surface and other platelets. The factors which attract other important cells to the injury are also released by the platelets. After that neutrophils are enter in the wound site. It used to fight against the infection, break downs the necrotic debris by attracting the macrophages and activates the fibroblast response.
The inflammatory phase remains 5 minutes to 5 days. After that it leads to the proliferation phase of the wound healing process. The primary platelet plug is formed by the aggregation of platelets. The platelets are activated by aggregation and attachment to the collagen surface. The degranulation, release of chemotactic and growth factors such as platelet-derived growth factor (PDGF), protease, and vasoactive agents are activated by enabling the platelet. In the 2 different pathways the coagulation occurs.
• Intrinsic pathway.
• Extrinsic pathway.
By activating the factor XII (Hageman factor) the intrinsic pathway begins. Whenever the blood is exposed to extravascular surface this factor is activated. Similarly, by activating the tissue factor the extrinsic pathway begins. Particularly the tissue factor found in the extravascular cells in the presence of factors VII and VIIa. Then both are proceeding with thrombin. The thrombin is then activated. It converts fibrinogen to fibrin. The fibrin is the important factor in the wound healing mechanism. The wound healing is impeded if the fibrin matrix is removed. As the result aggregation of platelet and coagulation cascade is clot formation. It occurs only in the site of injury.
In addition to activation of fibrin, thrombin facilitates migration of inflammatory cells to the sites of injury by increasing vascular permeability. By this mechanism, factors, and cells necessary to healing flow from the intravascular space and into the extravascular space (Figure 2).
Proliferation Phase
Proliferation phase is followed by inflammatory phase. It remains 24 to 72 hours. Mitotic activity is carried on the surface of the wound by bursting out the epidermal cells into mitotic activity. Across the surface of the wound, the cells begin to migrate. In the deeper parts of the wound the fibroblasts starts to proliferate. Small amounts of collagen are synthesized by these fibroblasts and it helps further fibroblast proliferation by acting it as a scaffold. Granulation tissue consists of capillary loops supported in this developing collagen matrix, also appears in the deeper of the wound. Fibroblastic phase is carried out in this phase. Depending upon the seviourity of injury, after four to five days, fibroblasts begins to produce large amounts of collagen and proteoglycans.
Collagen fibres are laid down randomly and are cross-linked into large, closely packed bundles. Proteoglycans are used to enhance the formation of collagen. In some injuries this phase remains up to few months. After 15 to 20 days of this phase it enters into maturation phase.
Maturation Phase
In this phase the collagen is remodeled into more organized manner. Tensile strength increased within one year depending on the seviourity of the wound. They may regain the original strength by 70 to 80%. After this phase the scar may formed.
While treating second and third degree burn wounds, pain causes during the dressing. To avoid that single time, use of natural scaffold is used to treat it. The scaffold is made from collagen rich fish skin. Collagen type-I plays a vital role in wound healing in the remodelling phase. Collagen type-I is abundantly available naturally in animals like fish. Particularly in fish bone, skin and tendon has high collagen. And it is easily available everywhere. So the fish skin is used as a scaffold. In this work the tilapia fish is used as a scaffold material. Because it is rich in collagen type-I when compared to other fishes. It is easily available fish. Its cost is less. Because of those reasons that fish is chosen. Indian plants have many number medicinal values. Tridax procumbens is one among that. It is commonly known as Jayanti Vedain Sanskrit and in English popularly called as ‘coat buttons’. The plant is native of tropical America and naturalized in Africa, Asia, Australia and India. The whole plant (weed, stem, leaf and flower) has a medicinal property. Especially have a wound healing property. It is distributed in all the areas, and available in all the seasons.
In this work the scaffold was prepared by sterilizing the tilapia’s skin and it is coated/blended with the tridax procumbens extract by using the laboratory instruments.
In the previous studies the scaffold is prepared only by sterilizing it by using specially designed equipment to it [1-5]. The collagen is derived to make a scaffold from the marine origin animals like fish because of its availability and cost. The derived collagen is especially used in wound healing application [5-10]. The herbs have its own medicinal property. And it is used for various medicinal applications. The tridax procumbens has the capability to heal the wound [5].
Methodology
(Figure 3)
Separation of tilapia’s skin
(Figure 4,5) Separation was done in a careful manner. Because removal of flesh after peeling is a difficult one. In the presence of flesh, the contamination may occur. The tilapia skins were peeled off which are the origin of Indian sea water. If it has any flesh it was removed by cutting. Then it was washed in the running distilled water for three to five minutes to avoid the traces of any blood or scale [10-14].
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