Comparison of the effect of guava ( psidium guajava l. ) Leaf extract x with polyvinylpyrrolidone-sodium hyaluronate on the number fibroblasts

Background : Wound healing is a cellular and complex process that aims to restore the integrity of tissue damaged by injury. The phases that occur in the wound healing process are inflammation, proliferation, and maturation (remodeling). One of the final results of wound healing is fibroblasts proliferation. Fibroblasts have the ability to synthesize collagen fibers that will cover the injured area. Natural medicinal plants that have the potential to accelerate wound healing are guava (Psidium guajava L.) leaves which contain flavonoids, tannins, saponins, and alkaloids. Polyvinylpyrrolidone-sodium hyaluronate gel is one of the wound treatment products on the market and can help the wound healing process Method: This research method was an experimental laboratory using 10 white rat samples which were divided into two groups and given injury to the mandibular anterior gingiva. Both treatment groups were applied topically in the morning and evening for 14 days, then the tissue was observed histologically. Results: Observation of the number of fibroblasts in the treatment group applied to guava leaf extract gel had a mean of 28±11 while the treatment group applied Polyvinylpyrrolidone-sodium hyaluronate gel was 10±4. The unpaired t test (Independent T-Test) showed a significance value of 0.000 (p<0.05) Conclusion: A significant difference in a comparative study of the effect of guava leaf extract with polyvinylpyrrolidone-sodium hyaluronate on the number of fibroblasts in gingival wound healing of Wistar rats.


INTRODUCTION
Wounds are defined as damage or loss of continuity in the skin or body tissues that can cause interference with the anatomical structure of the body. 1 Wounds can be caused by several things such as post-surgery, trauma, contact with heat sources, chemicals, and accidents. 2 Injuries that occur in the oral tissue one of which in the gingiva can occur due to surgery on the oral cavity. 3 Wound healing is a cellular and complex process that aims to restore the integrity of tissue damaged by injury and has several phases that are interrelated to one another. Phases that occur in the process of wound healing are the inflammatory phase, the proliferation phase, and the maturation phase (remodelling) . 4 In these phases there will be changes in components that can be observed microscopically such as leukocytes, fibroblasts, macrophages, collagen fibers and neurovascularization. 5 One determination of the end result of wound healing is fibroblast proliferation. Wound closure occurs because of collagen produced by fibroblasts, and the process of re-epithelialization in wound closure is the effect of the presence of fibroblasts. 6 One of the plants that has medicinal properties and is known by the public is guava (Psidium guajava Linn.). According to Soedibyo (1998), all parts of guava plants can be utilized and have properties including their leaves. 7 Guava leaf plants can be used as a wound healing because it has anti-inflammatory, hemostatic, and astringentic effects (cit. Soedibyo, 1998) . 7

Samples Preparation
This study used 10 male Wistar rats (Rattus norvegicus), ages 8-12 weeks, and weighed 200-300 grams obtained from the Faculty of Veterinary Medicine, Syiah Kuala University. 15 Rats were divided into 2 groups with each group totaling 5 rats.

Making Guava Leaf Extract
The sample used was guava leaf which was taken using a random sampling technique.

Preparation of Guava Leaf Extract Gel
The thick extract of guava leaves is processed into a gel preparation by dissolving with heated aquadest and then put into a mortar and added with Na-CMC, waiting until it expands then stirring for about 15 minutes until homogeneous and precipitated mucus forms down. The next process is to add propylene glycol, nipagin, glycerin, triethanol-amine, and stir until homogeneous. 17 Guava leaf extract put on another mortar and then dissolved using partially heated aquadest at 50˚C and added Na-CMC that has been formed, stirred until homogeneous until gel mass is formed.
The formed gel is stored in a vial bottle and left at room temperature overnight. 17,19 Animal Preparation

Polyvinylpirrolidone-sodium Hyaluronate Gel on Gingival Wounds
Wistar rats as many as 14 males were divided into two treatment groups namely, one treatment group was first applied to guava leaf extract gel topically and the second treatment group was applied polyvinylpyrrolidone-sodium hyaluronate gel. Both treatment groups were applied topically to the injured area evenly by using a 1 cc syringe as much as 0.1 ml each day twice in the morning and evening for 14 days. 7,21

Observation of Histological Preparations
After the treatment period ended, the rats were then deuthanated. The next process is tissue cutting in the mandible of the rats which is cut with a thickness of 2-3 mm and followed by a fixation

Data analysis
Data analysis was performed using SPSS 20 and hypothesis testing was performed with an unpaired T-test if the data distribution was normal with significant (p<0.05).

Observation Results on Histology Preparations
The results of this study indicate the average number of fibroblast cells in rat . The first treatment group applied guava leaf extract gel had an average number of fibroblasts of 28 ±11 while the second treatment group applied polyvinylpyrrolidonesodium hyaluronate gel had an average number of fibroblasts of 10±4 (Table 1).

Data analysis
Before performing the unpaired t test, used test data normality using the Shapiro-Wilk test because the sample size in this study is less than 50 samples.    given Carica papaya extract with a concentration of 75% than the control group with wound lengths that seem to start to close on day 3 and close by perfect on the 14th day and an increase in fibroblast cells, a perfect epithelial layer, and wound contraction. 15 Wound healing can be achieved by giving medicinal plants that contain material and nutrients to help speed up the wound healing process that is usually used in topical form. 28  .