The experiment and procedures used in this study was approved by the Institute of Animal Care Unit Committee of the Philippines (IACUCP), DMMMSU-NLUC.
The efficacy of the ointment on wound healing was tested in terms of its degree of redness, scab formation and duration of wound healing.
Table 4 presents the level of efficacy of the three (3) formulated treatments of Takip kohol (Centella asiatica) leaf ointment in terms of degree of redness, days for scab
formulation and duration of wound healing counted based on the number of days after the incision as compared the control drug mupirocin.
The table shows that in terms of degree of redness, treatment 3 has the same level of efficacy with the control drug which is very highly effective with an average value of 4.77 while treatments1 and treatment 2 have higher scale value of 4.92 but still are very highly effective. According to Lucas (1979), plant extracts help in circulation and skin problems and some of the cerebral circulatory and dermatological actions. The values shows that there is no significant difference on the three(3) treatments and the control drug in terms of degree of redness.
In the 3-stage model of wound healing, inflammation includes coagulation and inflammatory cell recruitment. The different clotting cascades are then initiated by clotting factors from the injured skin. The injured vessels follow a 5 to 10 minute
vasoconstriction, triggered by the platelets, to reduce blood loss and fill the tissue gap
with a blood clot comprised of cytokines and growth factors. Furthermore, the blood clot
contains fibrin molecules, fibronectin, vitronectin and thrombospondins, forming the provisional matrix as a scaffold structurse for the migration of leukocytes, keratinocytes, fibroblasts and endothelial cells and also acts as a reservoir of growth factors. The life-saving vasoconstriction with clot formation accounts for a local perfusion failure with a consecutive lack of oxygen, increased glycolysis and pH-changes. The vasoconstriction is then followed by a vasodilation in which the traumatized tissue suffers a reperfusion phenomenon. Both platelets and leukocytes release cytokines, chemokines and growth factors to activate the inflammatory process, stimulate the collagen synthesis, activate the transformation of fibroblasts to myofibroblasts, start angiogenesis and support the reepithelialization process. The vasodilatation can also be recognized by a local redness (hyperemia).https:/wound-healing.edu.
During proliferation, wound is rebuilt with ones granulation tissue which is comprised of collagen and extracellular matrix and into which new network of blood vessels develop into angiogenesis. Healthy granulation tissues is granular and uneven in texture, it does not bleed easily and in pink/red in color. The color and condition of the granulation tissue is often as indicator of how the wound is healing. http://www.clinimed.co.uk. Fibrolastic ( reconstructive) phase of repair begins 3 to 4 days after the injury and lasts up to 2 weeks ( Saladin, 2010).
The table shows the comparison of the number of days of scab formation on the incised wound of the subjects. As shown in the table, for the control drug, the average number of days before scab was formed is 2.17days, 2.5 days for T1, 2.17 days for T2 and 2.5 days for T3 respectively.
Epidermal development is responsible for scab formation, is a result of regeneration in treated mice confirmed that the extract had a positive effect towards cellular proliferation., granular tissue formation and epithelialization (Pawar, 2013).. There was a marked infiltration of the inflammatory cells, increased blood vessel formation and enhanced proliferation of cells as a result of the treatment with Takip kohol ointment.
Duration of Wound Healing
Table shows the comparison of the duration of wound healing in terms of the number of days from the day of the actual incision on the subjects. As shown in the table, the control drug, takes 6.2 days on the average to heal the wound, 7.2 days for T1, 5.2 days for T2 and 6 days for T3 respectively. The duration of wound healing of the three (3) concentrations of Takip kohol ointment and the control dug are comparable to each other as well as to the control drug and only further supported the latter finding.
Phenolic compounds present in the leaf extract promote the wound healing activity. (Sachan, 2011). Varied essential chemical found in the leaves further promote
its ability to kill bacteria whereby wounds are healed comparable with the standard antibacterial cream. This supports the report of Bushan ( 2013) that Takip kohol
promotes skin regeneration and strengthens the connective tissue and it assist with
cuts and wound healing,.
Alkaloids, tannins and flavonoids in the leaf extract are known to promote wound healing process and their anti-microbial property. Tannins is responsible for wound contraction and increased rate of epithelialization because of its astringent
property ( Surama and Nambat, 2013) . Its mechanism in wound healing is to stimulate
the production of type I collagen, which has an important role in wound closure and increases epithelialization of tissues. Flavonoids act by inhibiting the lipid peroxidation
process and are responsible for free radical scavenging, thus preventing and retarding
cell necrosis, and increasing vascularization at the wound site. Polyphenols, being compounds with antioxidant properties , also play a role in wound healing, through inhibition to lipid peroxidation(Miladiyah, 2012).
Proteins found are required as part of the inflammatory process in the immune response and in the development of granulation tissue. The main protein synthesized during the healing process is collagen, and the strength of the collagen determines wound strength ( http://www.dieteics.com. as cited by Picana, 2016).
Ethanolic extract of Takip Kohol (C. asiatica) leaf contains the phyto constitutents that promotes natural healing processes and could be used as a wound healing agent like other medicinal plants. These wound healing property may stimulate proliferation of fibroblasts, collagen formation, angiogenesis and reepithelialization (Budovsky, et al., 2015).
The Analysis of Variance (ANOVA) shows that there is no significant difference
among the three(3) treatments (T1, T2 & T3) of Takip kohol (Centella asiatica) leaf ointment and the control drug ( mupirocin) on wound healing in terms of degree of redness, scab formulation and the duration of wound healing. These implies that any of the formulated ointment concentration is as effective as the control drug and can be an alternative of the commercial ointment.
These findings confirm that the secondary metabolites found in Takip kohol leaf has pharmacological importance in human ailments, specifically of microbial in nature like wound.