The Beneficence from Gel of Ambon Banana (Musa Paradisiaca Var. Sapientum) to increase Wound Healing Post-Gingivectomy in Wistar Rat (Rattus Norvegicus)- In vivo Study

Diah, Rudhanton, Provisia M.Y. Wulan, Diona Olivia Yudianto

Faculty of Dentistry, Brawijaya University, Malang, Indonesia.

*Corresponding Author E-mail: wulanprovisia23@gmail.com

Abstract:

Periodontal dressing is the material used in post-surgical wounds. These dressings do not contain ingredients that can trigger healing, expensive and contains eugenol which can cause allergic reactions. Non-eugenol dressings are less able to adhere to the mucosa. Thus, it takes a natural dressing materials, is antibacterial, strong retention, cheap, and can accelerate healing. Ambon banana gel can be used for alternative of natural periodontal dressing because it contains several types of phytochemicals such as saponin, flavonoid, ascorbic acid and tanin which can accelerate the wound healing process. The purpose of this research is to test the effect of the application of banana gel to increase wound healing process in the injured area post gingivectomy on wistar rat (Rattusnorvegicus).The study begins by making extract of banana tree made into a banana gel in several doses, 50%, 75%, and 100% applied on 36 male Wistar rats (Rattus norvegicus) post-gingivectomy. The gel is applicated in experiment group for 1 week to determine the acceleration of healing compared with control group which were not given banana gel. In the control group maximum healing is on day-7, 50% is on day-5, 75% is on day-3, and 100% is the most effective increase healing process on day-1 (ANOVA, p=0,000) p<0,05.Conclusion of this research is Ambon banana gel can effectively improve the process of wound healing post-gingivectomy which increases acceleration of several variables include macrophages, neutrophils, lymphocytes, and capillary.

KEY WORDS: Ambon Banana Gel, Wound healing, Gingivectomy.

INTRODUCTION:

Periodontal disease is an oral health problem which has a relatively high prevalence in Indonesian society, with the prevalence of periodontal disease in all age groups is 96.58% (Tampubolon, 2010). One of periodontal disease is the enlargement of gingiva (gingival enlargement). Gingival enlargement is characterized by the addition of gingival size that can cause negative effects such as impaired function, both aesthetic function, the function of mastication, and occlusion function. Gingival enlargement can be corrected with gingivectomy procedure. Gingivectomy procedure leaves an open wound that had to be treated for post-treatment success. Healing after gingivectomy usually occurs within 2-4 weeks, and in the early post-gingivectomy used aid periodontal dressing for 1 week (Carranza, 2006). In the world of dentistry, periodontal material has been known to protect the wound dressing after gingivectomy. One such study conducted by Listyanti (2006) that sap Ambon banana trees (Musa paradisiaca var. Sapientum) used in the wound healing process using experimental animals mice showed satisfactory results. The study was followed by Bayu Febram (2010) to make Ambon banana stem sap into preparations ointment and applied in experimental animals mice to determine the wound healing process of the skin. The conclusion from these studies is the banana stem extract ointment preparations Ambon has an activity to accelerate the process of wound healing, accelerating infiltration of inflammatory cells, accelerate the process neokapilerisasi, reepitelisasi accelerate and improve the formation of connective tissue in the skin.

Wistar rats (Rattus norvegicus) is an animal that is easily available in large quantities, have a quick response, give an idea scientifically that may occur in humans and relatively cheap so it is widely used as a sample (Sihombing, 2010).The description above the underlying research on banana gum gel to accelerate the gingival wound healing after gingivectomy in vivo models of rats (Rattus norvegicus) by looking at the number of cells of neutrophils, macrophages, lymphocytes, and capillaries on injured area.

MATERIAL AND METHODES:

Research Design:

This study is an experimental research laboratory with a true experimental with posttest only control group design. This study aims to determine the effect of Ambon banana gel extract from Ambon banana tree (Musa paradisiaca var. Sapientum) in increasing the healing process in the gingival Wistar rats (Rattus norvegicus) post-gingivectomy were applied with Ambon banana gel (Musa paradisiaca var. sapientum) at the dose of 50%, 75% and 100%.

Gingivectomy Procedure:

The method used in gingivectomy procedure is using a round bur or a round diamond bur in Wistar rat’s anterior gingiva. Gingivectomy action taken is with a square-shaped design but not angled edges. Gingival wound size between approximately 1 cm x 0.5 cm. Effect of Ambon banana gel can be observed from the increasing the number of blood vessels or capillaries, macrophages, neutrophils and lymphocytes in the injured area after gingivectomy on days 1, 3 and 7. Then the results were compared between the control group. Further observations and analysis. The control group is the group of mice that were not given with Ambon banana gel, each treatment performed 3 repetitions.

Banana Gel Preparation and Application:

Ambon banana gel is a gel made with wet dispersion method. Making gel base begins with boiling water with water heater, while waiting for the water to boil, weigh all the ingredients in a measure Carbomer 0.519 g, 3.825 g Propylen glycol, Triethanolamine 1.275 ml, and 0.1275 g of Na-benzoate was dissolved in 3 drops of hot water. Upon completion of the manufacturing base of the trunk for sap mixing procedure of making banana gel with the dose of 50%, 75% and 100%. Gel is applied every day, and then the corresponding experimental group, performed surgery on day 1, 3 and 7 with exition of mandible and preparations made into histological preparations with pieces wound after gingivectomy with the longitudinal direction.

Measurement:

Measurement of the number of blood vessels, macrophages, neutrophils, and lymphocytes counted in the 10 field of view (magnification 400X) using a microscope and the percentage of wound healing by comparison between a matter of days and the number of capillaries, macrophages, neutrophils and lymphocytes between the control group to the treatment group.

RESULT:

The results of counting the number of blood vessels, macrophages, neutrophils and lymphocytes, in the control group, gel 50%, 75% and 100% on day 1, 3 and 7 is presented in figure (Figure 1) HE (Hematoksilin Eosin) M400x and Table below;

a b c d

Figure 1. Histology of gingiva post gingivectomy H+1, H+3, H+7

Figure 1 a-d. The Histology of gingiva post gingivectomy (HE M400x) day-1

a. Blood vessels, macrophages, neutrophils and lymphocytes P0 (Control)

b. Blood vessels, macrophages, neutrophils and lymphocytes P1 (Gel 50%)

c. Blood vessels, macrophages, neutrophils and lymphocytes P2 (Gel 75%)

d. Blood vessels, macrophages, neutrophils and lymphocytes P3 (Gel 100%)

e f g h

Figure 1 e-h. The Histology of gingiva post gingivectomy (HE M400x) day-3

e. Blood vessels, macrophages, neutrophils and lymphocytes P0 (Control)

f. Blood vessels, macrophages, neutrophils and lymphocytes P1 (Gel 50%)

g. Blood vessels, macrophages, neutrophils and lymphocytes P2 (Gel 75%)

h. Blood vessels, macrophages, neutrophils and lymphocytes P3 (Gel 100%)

i j k l

Figure 1 i-l. The Histology of gingiva post gingivectomy (HE M400x) day-7

i. Blood vessels, macrophages, neutrophils and lymphocytes P0 (Control)

j. Blood vessels, macrophages, neutrophils and lymphocytes P1 (Gel 50%)

k. Blood vessels, macrophages, neutrophils and lymphocytes P2 (Gel 75%)

l. Blood vessels, macrophages, neutrophils and lymphocytes P3 (Gel 100%)

Table 1. Mean and standard deviation from the number of blood vessels and Lymphocytes

Group		Blood vessel / capillary		Lymphocyte
Group		Mean	SD	Mean	SD
Day 1	Control	4,00	1,155	24,300	3,989
	Gel 50%	4,25	1,708	32,867	6,062
	Gel 75%	7,00	2,160	44,933	2,491
	Gel 100%	15,0	12,028	47,167	4,680
Day 3	Control	8,75	1,258	38,867	5,662
	Gel 50%	14,50	3,775	45,800	6,869
	Gel 75%	19,00	21,60	34,667	11,286
	Gel 100%	13,00	1,633	30,533	6,444
Day 7	Control	3,6	0,500	45,233	1,286
	Gel 50%	2,9	1,414	39,533	0,666
	Gel 75%	2,3	0,500	27,400	0,600
	Gel 100%	1,9	0,577	21,400	0,917

Table 2. Mean and standard deviation from the number of blood vessels and Lymphocytes

Group		Macrophage		Neutrophil
Group		Mean	SD	Mean	SD
Day 1	Control	19,666	1,528	28,600	2,5170
	Gel 50%	26,666	1,528	23,667	3,5105
	Gel 75%	16,666	1,528	18,367	3,1660
	Gel 100%	12,666	1,528	5,667	3,6005
Day 3	Control	24,333	3,512	11,467	0,3786
	Gel 50%	19,333	2,517	8,367	3,7207
	Gel 75%	11,333	1,528	6,233	1,8583
	Gel 100%	7,333	0,577	3,000	0,5292
Day 7	Control	18,666	1,528	8,433	0,2309
	Gel 50%	15,666	1,155	6,167	0,0577
	Gel 75%	8,333	0,55	4,567	0,2517
	Gel 100%	5,666	0,577	2,133	0,2082

From the results above, it can be seen that with increasing doses of banana gel then the sooner the healing process that is marked with the increasing number of blood vessels or capillaries, macrophages, neutrophils and lymphocytes. There are differences between the control group and the group in particular 100% gel. Test of normality and homogeneity of the data suggests that the data are normally distributed and homogeneous. One-way ANOVA test shows that the significance value of 0.000 (p<0.05), so it can be concluded that there are significant differences between treatment groups with the control group of the average number of capillaries. Pearson correlation test has strength correlation that there is a correlation between the use of Ambon banana gum gel with a dose of 50%, 75% and 100% with the direction of the positive correlation (correlation is positive).

DISCUSSION:

Ambon banana gel contains several types of phytochemicals that can accelerate the wound healing process, such as saponins, flavonoids, and tannins (Prasad et al., 2010). Saponins serves as hypocholesterolemic, immunostimulatory and anti-carcinogenic. Saponin anti-carcinogenic mechanisms include anti-oxidant and cytotoxic effect directly on cancer cells. Saponins also serves as an anti-bacterial (Almira, 2008). Saponins also have antifungal activity and defense against pathogenic microbes. (Khristyana et al., 2005). Flavonoids have anti-inflammatory effects, antioxidant, hypo-allergenic, and antiviral (Middletone et al., 2000). Tannins act accelerate wound healing and reduce scarring by inhibiting the formation and disposal of reactive oxygen substances. Another benefit of such tannins are analgesic, limiting secondary infection, prevent the loss of plasma fluid, and increase the proliferation of epithelial cells (Karodi et al., 2009).Increasing the number of blood vessel is caused by increasing the dose of banana gel, the content of active substances such as flavonoid, saponins, ascorbic acid and tannin also increased, active substances that have the effect to increase angiogenesis factor such as TGF (Transformer Growth Factor, AGF(Angiogenesis Growth Factor), FGF(Fibroblast Growth Factor) and VEGF(Vascular Endothelial Growth Factor). Increased angiogenes is factors can accelerate the proliferation of endothelial which form the walls of blood vessels resulting in increased amounts of blood vessels significantly in each treatment group. Active substances include saponins and flavonoids will increase aggregation and extravasation of monocytes so out of the intravascular lead to intravascular into macrophages and become in the way of phagocytosis against external agents that cause inflammation. On the first day after the injury, macrophages were in the area of inflammation because within a few minutes after inflammation begins and activates inflammatory products, macrophages that have been there and settled in tissue inflammation and tissue around the inflammation become activated and gather immediately start his response form the body's first line of defense and has the ability to split into macrophages more. However, supply of monocytes in the blood are few in number, thus requiring assistance from the bone marrow and causes macrophages increase with inflammation becomes chronic (Guyton, 2007). In the studies that have been done that by granting banana gum gel which contain saponins, flavonoids, and tannins may affect the amount of macrophages found in the injured area (Febram, 2008). Saponins contained in the banana gel can increase macrophage receptor (Febram 2008; Nengah, 2009), so that the movement of macrophages to the injured area to kill invasive organisms also increased (Kalsum et al, 2012). So that the healing of wounds will take place more quickly.

CONCLUSSION:

Conclusion is that applicating of Ambon banana gel with variant doses of 50%, 75%, and100% can affect the amount of gingival capillaries of male Wistar rats (Rattus norvegicus) post-gingivectomy. Correlation of Ambon banana gel to increase the number of capillaries is positive, an increasing the number of capillaries, macrophages, neutrophils and lymphocytes with increasing doses of Ambon banana gel.

ANKNOWLEDGMENTS:

Pathology-Anatomy Laboratory of Medical Faculty Brawijaya University, Malang, Indonesia.

Pharmacology Laboratory of Medical Faculty Brawijaya University, Malang, Indonesia.

Pharmaceutical Laboratory of Medical Faculty Brawijaya University, Malang, Indonesia.

REFERENCES:

1. Alzwar. 2009. Berkenalan dengan Aloe Vera, (Online), (http://www.azwar.web.ugm.ac.id/index.php?option=comcontent & task=view & id=7 & Itemid=2, diakses tanggal 8 Januari 2013).

2. Ashari. 2004. Holtikultura Aspek Budidaya. Jakarta: UI press.

3. Badan Penelitian dan Pengembangan Kesehatan. Riset Kesehatan Dasar (RISKESDAS) 2007. Kementrian Kesehatan Republik Indonesia: 2007.

4. Baratawidjaya KG. 2006. Reaksi Hipersensitivitas, Dalam: Imunologi Dasar, Edisi ke-7. Balai Penerbit FKUI. Jakarta, hal.157-161.

5. Bunner and Suddarth. 2002. Buku Ajar Keperawatan Medikal-Bedah. Jakarta: EGC.

6. Carranza FA, Newman M.G. 2002. Carranza’s Clinical Periodontology. 9th Edition, W.B. Saunders Elsevier Company, Philadelphia

7. Carranza, Fermin A., dkk. 2006. Clinical Periodontology 10th Edition. St. Louis: Saunders.

8. Desy, K. 2009. Laporan Praktikum, Sistem Organ Tikus Rattus norvegicus dan Pengamatan Sel Secara Mikroskopis. Blitar: Stikes Patria Husada.

9. Djulkarnain H.B. 1998. Pohon Obat Keluarga. Jakarta: Intisari.

10. EJ Caldeira, et al. 2007. Morphological Alterations In The Epithelium Of The Oral Mucosa Of Rats (Rattus norvegicus) Submitted to Long-term Systemic Nicotine Treatment. Elsevier. (online) http://www.aobjournal.com/article/ S0003-9969%2806%2900211-1/fulltext (diakses 20 Januari 2013, 13:35).

11. Eroschenko VP. 2003. Atlas Histologi di Fiore dengan korelasi Fungsional, Edisi 9, EGC, Jakarta, Hal. 29.

12. Falanga V. 2003. Mechanisms of Cutaneous Wound Repair, dalam: Freedberg IM, Wolff K, Eisen AZet al, editor, Fizpatrick’s Dermatology In General Medicine, Edisi ke-6, Graw-Hill, New York, p. 236-46.

13. Febram B, Wientarsih I, Priosoeryanto BP. 2010. Aktivitas Sediaan Salep Ekstrak Batang Pohon Pisang Ambon (Musa paradisiaca var. sapientum) dalam Proses Persembuhan Luka padaMencit (Musmusculusalbinus), Departemen Klinik, Reproduksi, dan Patologi, Fakultas Kedokteran HewanInstitut Pertanian Bogor.

14. Guyton, AC., Hall JE. 2007. Fisiologi Kedokteran, Edisi 11, EGC, Jakarta, Hal. 480-483.

15. Hanafiah, K.A. 2005. Rancangan Percobaan Aplikatif. Jakarta: PT Raja Grafindo Persada.

16. Harisaranraj R, Suresh K, Saravanababu S. 2009. Evaluation of the Chemical Composition Rauwolfia serpentina and Ephedra vulgaris, Advan Biol Res, 3(5-6): 174-8.

17. Harty, F.J and Ogston, R. 1995. Kamus Kedokteran Gigi, terj, alih bahasa drg. Narlan Sumawinat, Penerbit Buku Kedokteran, EGC, Jakarta.

18. Hasanoglu A, Ara C, Ozen S, Kali K, Senol M, Ertas E. 2001. Efficacy of Micronized Flavonoid Fraction in Healing of Clean and Infected Wounds, Int J Angiol,10: 41-4.

19. Indonesia Enterostomal Therapy Nurse Association (InETNA) and Tim Perawatan Luka dan Stoma Rumah Sakit Dharmais. 2004. Perawatan Luka, Makalah Mandiri, Jakarta.

20. Ismail. 2008. Luka dan Perawatannya. Merawat luka. (online) http://images.mailmkes.multiply.multiplycontent.%20luka.pdfnmid. (diakses tanggal 18 Oktober 2012, 07:10).

21. Jayanto, Galih. 2012. Wound Healing / Penyembuhan Luka, (Online), (http://doktercilix.blogspot.com/2012/11/wound-healing-penyembuhan-luka.html, diakses 27 Desember 2012), FK UII Yogyakarta.

22. Listyanti A.R. 2006. Pengaruh Pemberian Getah Bonggol Pisang Ambon (Musa paradisiaca var. sapientum) dalam Proses Penyembuhan Luka pada Mencit (Mus musculus albinus). [Skripsi]. Fakultas Kedokteran Hewan IPB. Bogor.

23. MacKay D, AL Miller. 2003. Nutrional Support for Wound Healing, Altern Med Rev, 8(4): 359-375.

24. MansjoerA, dkk. Eds.2000.Kapita Selekta Kedokteran. Edisi III, Media Aesculapius FKUI, Jakarta.

25. Marimuthu L. 2009. Tumbuh-tumbuhan Beracun, (Online),(http://pkukmweb.ukm.my/~ahmad/tugasan/s2_99/a72826.htm, diaksestanggal 8 Januari 2013).

26. Morison, Moya J. 2003. Manajemen Luka. Jakarta: EGC.

27. Muntiha M. 2001. Teknik Pembuatan Preparat Histopatologi dari Jaringan dengan Pewarnaan Hematoksilin dan Eosin (HE), Temu Teknis Fungsional Non Peneliti, Balai Penelitian Veteriner, Bogor.

28. Nowak, R. 1983. Walker’s mammals of the World, 4th Edition. Baltimore, MD: The johns Hopkins University Press.

29. Oktaviyanti. 2004. Kajian Penggunaan Gel Lidah Buaya (Aloe Vera) pada Pembuatan Hand and Body Cream. [skripsi]. Bogor: Fakultas Teknologi Pertanian, Institut Pertanian Bogor.

30. Oudoff MJ, Bolscher JGM, Nazmi K. 2008. Histatins are the major wound-closure stimulating factors in human saliva as identified in cell culture assy, (Online), (http://www.fasebj.org/content/22/11/3805.full.pdf, diakses tanggal 1 Januari 2013).

31. Petter, F. 2000. Silabus Periodonti. Terjemahan oleh Amaliya. 2004. Jakarta: EGC.

32. Pratiwi M. 2011. Efek Ekstrak Lerak (Sapindus rarak DC) terhadap Penurunan Sel-sel Radang Pada Tikus Wistar Jantan (Penelitian In Vivo). Paper. Tidakditerbitkan, Fakultas Kedokteran Gigi Universitas Sumatera Utara.

33. Robin, et al. 1995. Granzyme B Plays a Critical Role in Cytotoxic Lymphocyte-induced Apoptosis.Imunological Review Volume 146. Issue 1. p.211-221.

34. Rosen, et al. 2008. On The Comparison of Parametris and Non Parametric Boostrap. UUDM Report. Sweden: Uppsala University.

35. Sugiyono. 2009. Metode Penelitian Kuantitatif, Kualitatifdan R and D. Bandung: Alfabeta.

36. Sihombing M, Raflizar. 2010. Status GizidanFungsi Hati Mencit (Galur CBS-Swiss) dan Tikus Putih (Galur Wistar) di Laboratorium Hewan Percobaan Puslitbang Biomedisdan Farmasi, Media Litbang Kesehatan Volume XX Nomor 1 Tahun 2010, Badan Litbangkes Puslitbang Biomedis dan Farmasi, Jakarta.

37. Smith Bj, Mangkoewidjojo S. 1988. Pemeliharaan, Pembiakan dan Penggunaan Hewan Cobaan di Daerah Tropis. Universitas Indonesia Press. Jakarta.

38. Sunarjono H. 2007. Budidaya Pisang dengan Bibit Kultur Jaringan. Jakarta: Penebar Swadaya.

39. Suriadi. 2004. Perawatan Luka. Jakarta: Sagung Seto.

40. Suyanti. 2008. Pisang, Budidaya, Pengolahan dan Prospek Pasar. Edisi Revisi. Jakarta: penebar Swadaya.

41. Tampubolon, Nurmala Situmorang. Dampak Karies Gigi dan Penyakit Periodontal Terhadap Kualitas Hidup, (Online), (http://library.usu.ac.id, diakses 20 Desember 2012).

42. Taylor C, et al. 1997. Fundamental of Nursing, Lippincott Raven, Washington.

43. Tizard I. 1982. Veterinary Immunology. An Introduction, Ed ke-3. Saunders WB co, Masduki Partodiredjo, penerjemah, 1988, Airlangga University Press, Surabaya.

44. Topazian RG, Goldberg MH, and Hupp JR. 2002. Oral Maxillofacial Infection. 4^thEd., WB Saunders, USA, p. 25.

45. Wijaya. 2010. Getah Pisang sebagai Obat Alternatif tradisional Penyembuh Luka Luar menjadi Produk Industri. Bogor Agricultural University.

46. Wilmana F.P. 2007. Analgesik-AntipiretikAnalgesikAntiinflamasiNonsteroisdanObatGnagguanSendiLainnya, dalam Farmakologidan Terapi, Editor Ganiswara, S.G, Edisi ke-5. Tidak Diterbitkan, Farmakologi Fakultas Kedokteran Universitas Indonesia, Jakarta.

47. Yosaphat, dkk. 2010. Getah Batang Pisang Raja (Musa Sapientum) untuk Sembuhkan Luka Setelah Pencabutan Gigi, (Online), (http://forum.kompas.com/alternatif/32006-getah-pisang.html, diakses 27 Desember 2012), FKG UGM, Yogyakarta.

Received on 05.11.2018 Modified on 20.12.2018

Research J. Science and Tech. 2019; 11(2):89-93.

DOI: 10.5958/2349-2988.2019.00014.7