<p>Potensi ekstrak daun singkong (Manihot esculenta Crantz) terhadap ekspresi MMP-8 fibroblas gingiva pada model tikus dengan disfungsi ovarium dan periodontitis</p><p>Potential of cassava (Manihot esculenta Crantz) leaf extract on the MMP-8 expression of gingival fibroblast in rats model with ovarian dysfunction and periodontitis</p>

Abstract

Pendahuluan: Disfungsi ovarium merupakan kondisi yang menimbulkan defisiensi hormon estrogen dan progesteron. Penurunan hormonal ini menyebabkan peningkatan produksi tumor necrosis factor (TNF) yang dapat memicu peningkatan produksi matriks metalloproteinase-8 (MMP-8). Periodontitis yang disebabkan oleh bakteri gram negatif akan memicu makrofag melepaskan TNF-α yang berkontribusi dalam pembentukan MMP-8. MMP-8 ini berperan dalam degradasi kolagen jaringan ikat gingiva. Meningkatnya MMP-8 dapat menyebabkan terjadinya periodontitis. Gejala periodontitis karena disfungsi ovarium dan induksi bakteri ini dapat diperlambat dengan bahan alam, yaitu daun singkong. Tujuan penelitian ini adalah untuk menganalisis potensi ekstrak daun singkong (Manihot esculenta Crantz) terhadap ekspresi MMP-8 fibroblas pada model tikus disfungsi ovarium dan periodontitis. Metode: Penelitian ini merupakan penelitian eksperimental laboratoris dengan post-test only control group design. Sampel yang digunakan adalah tikus Sprague-Dawley betina yang dibagi dalam 5 kelompok yaitu (1) kelompok kontrol (K), (2) Kelompok yang diinduksi bakteri Porphyromonas gingivalis dan diberi aquades, (3) Kelompok yang diinduksi bakteri Porphyromonas gingivalis dan diberi ekstrak daun singkong, (4) Kelompok yang diberi perlakuan ovariektomi dan diberi aquades, (5) Kelompok yang dilakukan ovariektomi dan diberi ekstrak daun singkong. Pengambilan jaringan gingiva setelah tahap euthanasia dilakukan untuk pembuatan preparat histopatologi dengan pewarnaan imunohistokimia. Pengamatan dan penghitungan ekspresi MMP-8 dilakukan dengan menggunakan software ImageJ dan Immunoratio. Hasil: Hasil analisis data one-way ANOVA, ekspresi MMP-8 fibroblas gingiva menunjukkan perbedaan yang signifikan (p=0,000). Hal ini menunjukkan bahwa ekstrak daun singkong dapat menurunkan ekspresi MMP-8 fibroblas gingiva tikus yang mengalami disfungsi ovarium dan periodontitis diinduksi P. gingivalis. Simpulan: Ekstrak daun singkong (Manihot esculenta Crantz) dapat menurunkan ekspresi MMP-8 sel fibroblas gingiva pada model tikus disfungsi ovarium dan periodontitis.Kata kunci: Disfungsi ovarium, periodontitis, ekspresi MMP-8, ekstrak daun singkong. ABSTRACTIntroduction: Ovarian dysfunction is a condition that causes estrogen and progesterone deficiency. This hormonal decrease causes an increase in the production of Tumour Necrosis Factor (TNF), which can trigger an increase in the production of matrix metalloproteinase-8 (MMP-8). Periodontitis caused by gram-negative bacteria will trigger macrophages to release TNF-α, which contributes to the formation of MMP-8. MMP-8 plays a role in collagen degradation of the gingival connective tissue. An increase in MMP-8 can cause periodontitis. Periodontitis symptoms due to ovarian dysfunction and bacterial induction can be slowed down by natural ingredients, such as cassava leaf. The purpose of this study was to analyse the potential of cassava leaf extract (Manihot esculenta Crantz) on MMP-8 expression of gingival fibroblast in rats model with ovarian dysfunction and periodontitis. Methods: This study was an experimental laboratory with post-test only control group design. The samples were female Sprague-Dawley rats divided into five groups: (1) control group (K); (2) Porphyromonas gingivalis induced group and given aquadest; (3) Porphyromonas gingivalis induced and given cassava leaf extract; (4) Group with ovariectomy treatment and given aquadest; (5) Group with ovariectomy treatment and given cassava leaf extract. Gingival tissue retrieval after the euthanasia was carried out for the histopathology preparations by immunohistochemical staining. Observation and calculation of MMP-8 expressions were performed using ImageJ and Immunoratio software. Results: The results of the one-way ANOVA analysis of MMP-8 expression of gingival fibroblasts showed a significant difference (p = 0.000); thus cassava leaf extract reduce the MMP-8 expression of gingival fibroblasts of rats with ovarian dysfunction and P. gingivalis induced periodontitis. Conclusion: Cassava (Manihot esculenta Crantz) leaf extract can reduce the MMP-8 expression of gingival fibroblast cells in rats’ model with ovarian dysfunction and periodontitis.Keywords: Ovarian dysfunction, periodontitis, MMP-8 expression, cassava leaf extract.

Increasing fibroblasts and gingival collagen density in periodontitis rats by using cassava leaf extract

“…Decreased levels of these cytokines will speed up the healing process and increase fibroblasts quantity and collagen density. 29 Another study showed that surgically induced skin wounds in rats could be successfully treated with cassava leaf extract. Arranging connective tissue in a parallel pattern to collagen fibers accelerated the remodeling phase by several steps.…”

Superoxide Dismutase Expression in the Ovaries of Periodontitis Animal Models Induced by Porphyromonas gingivalis and Treated with Cassava Leaves Extract

“…Cassava leaves extract containing bioactive compounds might have antioxidant activity. Our previous study revealed that the active ingredients, such as vitamins C, E, polyphenols, and flavonoids, were identified in the cassava leaf extract [15]. The cassava leaves extract acting as an exogenous antioxidant might balance the excessive oxidants caused by P. gingivalis infection.…”

“…It was stirred twice every 24 hours. The solution was concentrated with a rotary evaporator at 50 0 C and 90 rpm until the solution was semi-solid [15].…”

“…For the periodontitis model, the animal models were injected with 5mg/ 0.5 ml PBS (Phosphate Buffer Saline) of P. gingivalis, in which the concentration was 2.10 9 CFU/ml, in the buccal and lingual gingival grooves of the lower left first molar. The injection was administered every three days for two weeks using a 30G tuberculin syringe [15]. After 14 days, the animal models (T2) were treated with 179.2 mg/kg BW cassava leaves extract orally, divided into two administrations (2ml every administration at 8.00 AM and 8.00 PM) daily for a week [15].…”

“…The injection was administered every three days for two weeks using a 30G tuberculin syringe [15]. After 14 days, the animal models (T2) were treated with 179.2 mg/kg BW cassava leaves extract orally, divided into two administrations (2ml every administration at 8.00 AM and 8.00 PM) daily for a week [15]. Likewise, T3 and T4 were treated with vitamin C and metronidazole, respectively.…”