Annals of Biology 36 (2) : 220-226, 2020 Effects of Centella asiatica Extract on Pro-inflammatory Cytokines (TNF-α) in Severe Early Childhood Caries and Caries Free PRIYAWAN RACHMADI, MUHAMMAD LUTHFI 1, *, AQSA SJUHADA OKI 1 , MIEKE SYLVIA MAR 2 AND MUHAIMIN RIFAI 3 Department of Dental Material, Faculty of Dentistry, Universitas Airlangga, Surabaya 60285, Indonesia *(email : m.luthfi@fkg.unair.ac.id, Phone : +6281357898957) ABSTRACT Dental caries is a chronic, multifactorial disease that begins with a microbiological shift in complex biofilms (dental plaque) that is influenced by consumption of sugar, salivary flow, and behaviour. The present investigation was undertaken to analyze the expression of pro-inflammatory cytokines TNF-α on salivary neutrophil surfaces using flow cytometer. Extra Centella asiatica reduced pro-inflammatory cytokines TNF-α on the neutrophil surface of salivary free caries at 0 µg/ml (24.04±0.70), 12.5 µg/ml (21.88±1, 48), 25 µg/ml (20.56±0.95), 50 µg/ml (19.28±1.32), 100 µg/ml (18.07±1.17), 200 µg/ml (17.28±0.84), while in S-ECC at 0 (35.95±3.50), 12.5 (28.88±1.32), 25 (27.82±1.28), 50 (26.46±2.95) and 100 (23.46±1.42). C. asiatica decreased pro-inflammatory cytokines TNF-α produced by salivary neutrophils. Key words : Severe early childhood caries, neutrophil cells, TNF-α pro-inflammatory cytokines 1 Department of Oral Biology, Faculty of Dentistry, Universitas Airlangga, Surabaya 60285, Indonesia. 2 Departement of Forensic Odontology, Faculty of Dentistry, Universitas Airlangga, Surabaya 60285, Indonesia. 3 Department of Physiology, Cell Culture and Animal Development, Universitas Brawijaya, Malanag 65145, Indonesia. INTRODUCTION One of the most common diseases in children around the world is caries early childhood (ECC), this disease not only affects the oral health of early childhood but affects general health (Naidu et al., 2016), with signs that there is a smooth surface decay on children under three years of age and usually begins as soon as the first tooth erupts and develops rapidly into cavitation only in 6 to 12 months. This disease will cause a decrease in the quality of health for children, because ECC can reduce the ability to chew on children so that the impact can disrupt the growth and development of children in general (Sun et al., 2016). Demineralization of tooth enamel is caused by acidogenic bacteria that ferment carbohydrate- containing foods. After carbohydrates are digested, especially sucrose, there will be a rapid decrease in pH in the biofilm attached to the teeth to 5.0 or below. Low pH causes dysbiotic micro biome which is characterized by an increase in the proportion of acid biofilm species and changes in the composition of the biofilm matrix (Tinanoff et al., 2019). Streptococci mutants (SM) bacteria are the main bacteria as a cause of ECC. In the last decade, several studies were carried out regarding the relationship between caries occurrence and the number of BC in saliva (Kawashita et al., 2011). Neutrophils are very important cells in the host defence process against pathogenic microbial attacks. Neutrophils have the ability to phagocytosis and kill microbes because neutrophils are the cells that have the most bactericidal capacity by using oxygen- dependent and oxygen independent processes (Scott et al., 2018). The killing of pathogenic microbes that attack the host is mediated by professional phagocytes, namely, neutrophils through phagocytosis by microbial opsonisation because neutrophils express many receptors used to recognize microbes by binding to their specific molecules and host proteins (such as IgG and complement) (Rigby and DeLeo, 2012). In addition, neutrophils have been shown to produce various cytokines, chemokine and growth factors thus becoming a major contributor in the production of pro- inflammatory cytokines in the area of infection (Mantovani et al., 2011). Children with ECC have high pro-inflammatory