Browsing by Author "Rohazila Mohamad Hanafiah"
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Publication Anti-Bacterial Activities of Melastoma Malabathricum Stem Bark Fractions against Streptococcus Mutans(Universiti Sains Islam Malaysia, 2018) ;Rohazila Mohamad Hanafiah ;Siti Aisyah Abd Ghafar ;Zurairah Ibrahim ;Wan Yaacob Wan Ahmad ;Wan Syaidatul AqmaNazlina IbrahimThe objective of this study is to isolate and identify the bioactive compounds that possess antibacterial activities from Melastoma malabathricum stem bark acetone extract (MMSBAE) against Streptococcus mutans. M. malabathricum is widely used in the Southeast Asia to treat many ailments. A total of 12 fractions was purified by vacuum liquid chromatography (VLC) and further analysed by TLC-bioautography to determine antibacterial activities. TLC-bioautography showed that fraction 9 possesses antibacterial activities against S. mutans. Identification of fraction 9 had been done by GCMS and revealed 21 compounds. Some of the compound were important as agent pharmaceutical such as α-amyrin, β-sitosterol, hexadecenoic acid, stearic acid and hexacosanoic acid. Crystal violet and glass surface assay were used to determine anti-biofilm and anti-adherence activity, respectively. The concentrations of fraction 9 that produce 50% reduction in anti-biofilm and anti-adherence activities were 5 mg/mL and 2.50 mg/mL, respectively. Scanning electron microscopy (SEM) was performed to visualize the effect of the fraction 9 on biofilm structure of S. mutans. SEM analysis showed lysed biofilm were found on treated cells. These results indicated that this fraction possesses a powerful anti-cariogenic potential against S. mutans. - Some of the metrics are blocked by yourconsent settings
Publication Anti-biofilm and anti-adherence activities of Spilanthes acmella leaves extracts towards Streptococcus mutans(Frontiers, 2018) ;Rohazila Mohamad Hanafiah ;Siti Sarah Azhar ;Luqman Atif EliasSiti Aisyah Abd GhafarBackground: Streptococcus mutans is the main causative agents of dental caries by developing biofilm and increase adherence activity. Dental caries have becoming more common due to antibiotic resistance towards Streptococcus mutans. Spilanthes acmella is well-known as anti-toothache plant with high medicinal usages. It has been recognized as an important medicinal plant with high demand worldwide. Therefore, the objective of this study is to establish antimicrobial activities, including anti-biofilm and anti-adherence activity of Spilanthes acmella on cariogenic Streptococcus mutans. Methods: Spilanthes acmella leaves extracts were prepared by serial extraction with n-hexane, dichloromethane, acetone and methanol. Antibacterial activities of these extracts were conducted using disc diffusion assay, anti-biofilm and anti-adherence activities methods. Results: The result indicates that n-hexane and methanol leaves extracts are endowed with anti-streptococcus activity. Both methanol and n-hexane extracts inhibit moderately Streptococcus mutans growth as indicated in the disc diffusion assay. Methanol and a n-hexane extracts inhibit the biofilm formation and adherence activity of Streptococcus mutans. Conclusion:This study highlights the potential of Spilanthes acmella leaves extract as a new promising anti Streptococcus mutans agents. - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial Activity of Bioactive Compound in Salvadora persica (Chewing Stick) Against Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans(ASM SCIENCE JOURNAL, 2022) ;Haslinda Ramli ;Zalina BaharudinRohazila Mohamad HanafiahThis study aims to determine the phytochemical properties of Salvadora persica extracts and their antibacterial activities against Gram-negative oral anaerobes which are responsible of periodontitis such as Porphorymonas gingivalis and Aggregatibacter actinomycetemcomitans. Aqueous and ethanolic extracts were prepared using the root powdered stem of S. persica. The phytochemical compounds were determined with Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography-Mass Spectrometry (GC-MS). The antibacterial activities were assessed according to the levels of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The S. persica ethanolic and aqueous extracts contained antibacterial compounds such as Octadecenoic acid, methyl ester, n-Hexadecanoic acid and Pentadecanoic acid, 2,6,10,14-tetramethyl-, methylester, 1-(2,2-Dimethylpropanoyl)-L-prolyl-L-prolyl-N,2-dimethylalaninamide and 2-({4- [(Difluoromethyl)sulfanyl]phenyl} amino) benzoic acid, respectively. Both P. gingivalis (0.025 mg/mL) and A. actinomycetemcomitans (0.125 mg/mL) were susceptible to S. persica ethanolic extract. Ethanolic extract preparation has equal antibacterial activity with 0.2% chlorhexidine gluconate solution (p<0.05). Ethanolic extract (0.025-0.125 mg/mL) preparation has comparable require biocompatible testing - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial activity of silver nanoparticles Clinacanthus nutans (Agnp-CN) against Streptococcus mutans(Frontiers Media S.A, 2018) ;Siti Aisyah Abd Ghafar , ;Nor Baitie Adura Muhd Fudzi ;Wan Nur Farhanah Wan Sulaiman ;Rohazila Mohamad HanafiahVuanghao LimClinacanthus nutans was found to possess anti-venom, anti-inflammatory, analgesic, anti- diabetic, anti-rheumatism, antiviral and antioxidant properties. Silver nanoparticles are nanoparticles between 1nm to 100nm in size and play significant role in medicinal fields. Silver nanoparticles exhibit distinctive properties, such as good conductivity, chemical stability, catalytic and antibacterial activity. Streptococcus mutans is commonly found in human oral cavity and the main contributor to tooth decay. There is no study on antibacterial effect of silver nanoparticles Clinacanthus nutans (AgNp-CN) against Streptococcus mutans reported to date. Therefore, objective of this study is to investigate antibacterial properties of silver nanoparticles Clinacanthus nutans against Streptococcus mutans. - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial Activity Of Spilanthes Acmella Flower Extracts (safe) Against Streptococcus Mutans(UPM, 2019) ;Siti Aisyah Abd Ghafar ;Muhammad Fikhry Mohd Salehuddin, ;Nur Syamimi Syuhada Che AwangRohazila Mohamad HanafiahIntroduction:Spilanthes acmella, also known as “subang nenek’, has been used traditionally in Malaysia to treat toothache. A previous study has shown Spilanthes acmella leaves extracts (SALE) inhibit Streptococcus mutans growth. Streptococcus mutans is commonly found in the human oral cavity and is the main contributor to tooth de-cay. There is no study on the antibacterial effects of Spilanthes acmella flower extracts (SAFE) against Streptococcus mutans reported to date. Therefore, the objective of this study is to investigate antibacterial properties of SAFE against S. mutans. Methods:S. mutans was subcultured in Muller Hinton (MH) broth and agar. Sequential extractions of S. acmella flowers were conducted using four different solvents with increasing polarity, [n- hexane, dichloromethane (DCM), acetone, methanol (MeoH)] and tested with different concentrations against S. mutans via the disc diffusion assay, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Sodium fluoride (NaF) was used as a positive control while DMSO was used as a negative control. Results: The disc diffusion assay shows SAFE inhibited Streptococcus mutans growth. SAFE-DCM shows the greatest inhibition properties (12.33±2.30 mm) followed by SAFE-n-hexane (11.33±0.57 mm). Meanwhile, SAFE-Meoh and SAFE-acetone show no inhibition zone (6.00±0.001 mm). MIC value for SAFE-DCM and SAFE-n-hexane is 12.5 mg/mL respectively. Whereas, MBC value SAFE-DCM and SAFE-n-hexane is 50.0 mg/mL respectively. Conclusion: It can be concluded SAFE-DCM and SAFE-n-hexane possesses bactericidal properties against Streptococcus mutans. - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial And Antibiofilm Analyses Of Melastoma Malabathricum Leaves Extract Against Streptococcus Mutans On Tooth Surfaces(Malaysian Society for Microbiology, 2020) ;Fatin Farhana Azizan ;Rohazila Mohamad Hanafiah ;Marwan Jawad Msarah ;Norefrina Shafinaz Md. Nor ;Nazlina IbrahimWan Syaidatul AqmaAims: Dental caries is a chronic infectious disease caused by Streptococcus mutans due to its ability to form biofilm. This study aims to assess the antimicrobial efficacy of Melastoma malabathricum leaf extract against S. mutans on the surface of tooth samples as a potential therapy for dental caries. Methodology and results: Extraction of M. malabathricum leaves was done using acetone as the solvent and antibacterial activity of the extracts was determined by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Antibiofilm activity of M. malabathricum extract against S. mutans was determined by comparing the colony count, biofilm formation assay and morphology observation by scanning electron microscope (SEM). The MIC value of extracts was 6.25 mg/mL and MBC value was >25 mg/mL. A decrease in colony count was noted when tooth samples were incubated with M. malabathricum extract for 8 h compared to 4 h incubation. At pH 5, the formation of the colony was the least, medium at pH 8 and maximum at pH 7. A decrease in biofilm formation was observed when tooth samples were incubated with the extract for 8 h. SEM observations showed treatment with the extract caused S. mutans cell membrane to leak leading to cell morphology changes. Conclusion, significance and impact of study: Acetone extract of M. malabathricum leaves showed excellent antibacterial activity against S. mutans. It has bactericidal activity with the ability to inhibit biofilm in dose-dependent manner against S. mutans. The morphological analyses suggested that the extract disrupted the cell membrane of the bacteria. - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial and biofilm inhibition activities of Melastoma malabathricum stem bark extract against Streptococcus mutans(Universiti Sains Malaysia, 2015) ;Rohazila Mohamad Hanafiah ;Wan Syaidatul Aqma ;Wan Ahmad Yaacob ;Zulfahmi Said ;Nazlina Ibrahim ;Faculty of Dentistry ;Universiti Kebangsaan Malaysia (UKM)Universiti Sains Islam Malaysia (USIM)Aim: This study was to determine the antibacterial activity of Melastoma malabathricum stem bark acetone extract (MMSBAE) against Streptococcus mutans. Methodology and results: Antibacterial activity of the extract was determined by minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), biofilm formation, adherence inhibition, time kill studies and effect on S. mutans membrane integrity. MIC and MBC values of MMSBAE were 1.25 and 5 mg/mL, respectively. Time kill studies showed that reduction of colony forming unit in treated cells is 3 log10 after 10 h of treatment (p < 0.05). The extracts reduced 50% biofilm and adherence activity of S. mutans at 1.88 mg/mL. The effect on S. mutans membrane integrity after exposure to MMSBAE for 90 and 120 min was determined by measuring leakage of cell content at 2 different wavelengths of 260 nm and 280 nm. In leakage assay, the percentage of absorbance (260 nm) in treated cell material showed 57% at 90 min and 60% at 120 min which is higher than negative control (<20%) but less than positive control (100%). The percentage absorbance of treated cell material (280 nm) was 61% at 90 min and 63% at 120 min. Identification of compound in MMSBAE was done by gas chromatography mass spectrometry (GCMS). Ten compounds were identified in the MMSBAE with some of them important in antimicrobial activity such as ethyl ester, undecene, and gamma sitosterol. Conclusion, significance and impact of study: MMSBAE showed excellent bactericidal and antibacterial activities against S. mutans. The antibacterial mode of action of MMSBAE is suggested to be the disruption of the S. mutans membrane structure. The MMSBAE significantly inhibited biofilm and adherence activities of S. mutans in dose dependent manner (p < 0.05). MMSBAE has potential in the development of antibacterial agent with anti-biofilm and anti-adherence activities. - Some of the metrics are blocked by yourconsent settings
Publication Antibacterial properties of clinacanthus nutans extracts against porphyromonas gingivalis and aggregatibacter actinomycetemcomitans: An in-vitro study(University of Dicle, 2019) ;Rohazila Mohamad Hanafiah ;Khairine Alia Che Kamaruddin ;Nurul Amira Ahmad Saikin ;Wan Nur Alwani Binti Wan Abdul Aziz ;Muhammad Fahmi Yakop ;Vuanghao Lim ;Siti Aisyah Abd Ghafar ;Nuramirah Azizan ;Shahida Mohd Said ;Faculty of Dentistry ;Universiti Sains Islam Malaysia (USIM) ;Universiti Sains Malaysia (USM)Universiti Kebangsaan Malaysia (UKM)The aim of this study is to futher investigate and validate the antibacterial effect of Clinacanthus nutans plant extract against periodontal pathogens namely Porphyromonas gingivalis and Aggregatibacter actinomycetamcomitans. Four samples of alcoholic extract of C. nutans leaves were used in different concentrations i.e. 100%, 50%, 10% ethanol and 100% chlorofom and Chlorhexidine 0.2% (CHX) was used as the positive control. The antibacterial activity of C.nutans extract were investigated using disc diffusion agar test for determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). In this study, 50% ethanol extract and 100% chlorofom extract were found to have antibacterial activity against P.gingivalis, while only 50% ethanol crude extract was found to have acceptable antibacterial activity against A. actinomycetamcomitans (p < 0.05). The MIC and MBC tests showed that 50% ethanol extract had bacteriostatic activity against both P.gingivalis and A. actinomycetamcomitans while 100% chlorofom extract had bactericidal activity against P. gingivalis. These two findings were also found to be better than the activity of CHX. C. nutans extract was found to have notable antibacterial activity against P. gingivalis and A. actinomycetamcomitans comparable to CHX 0.2%. � 2019 Journal of International Dental and Medical Research. - Some of the metrics are blocked by yourconsent settings
Publication Critical Evaluation of Green Synthesized Silver Nanoparticles-Kaempferol for Antibacterial Activity Against Methicillin-Resistant Staphylococcus aureus(Taylor & Francis, 2024) ;Ariff Haikal Hairil Anuar ;Siti Aisyah Abd Ghafar ;Rohazila Mohamad Hanafiah ;Vuanghao LimNur Farah Atiqah Mohd PazliIntroduction: This study aimed to characterize silver nanoparticles-kaempferol (AgNP-K) and its antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA). Green synthesis method was used to synthesize AgNP-K under the influence of temperature and different ratios of silver nitrate (AgNO3 and kaempferol). Methods: AgNP-K 1:1 was synthesized with 1 mM kaempferol, whereas AgNP-K 1:2 with 2 mM kaempferol. The characterization of AgNP-K 1:1 and AgNP-K 1:2 was performed using UV–visible spectroscopy (UV–Vis), Zetasizer, transmission electron microscopy (TEM), scanning electron microscopy-dispersive X-ray spectrometer (SEM-EDX), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. The antibacterial activities of five samples (AgNP-K 1:1, AgNP-K 1:2, commercial AgNPs, kaempferol, and vancomycin) at different concentrations (1.25, 2.5, 5, and 10 mg/mL) against MRSA were determined via disc diffusion assay (DDA), minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) assay, and time-kill assay. Results: The presence of a dark brown colour in the solution indicated the formation of AgNP-K. The UV–visible absorption spectrum of the synthesized AgNP-K exhibited a broad peak at 447 nm. TEM, Zetasizer, and SEM-EDX results showed that the morphology and size of AgNP-K were nearly spherical in shape with 16.963 ± 6.0465 nm in size. XRD analysis confirmed that AgNP-K had a crystalline phase structure, while FTIR showed the absence of (-OH) group, indicating that kaempferol was successfully incorporated with silver. In DDA analysis, AgNP-K showed the largest inhibition zone (16.67 ± 1.19 mm) against MRSA as compared to kaempferol and commercial AgNPs. The MIC and MBC values for AgNP-K against MRSA were 1.25 and 2.50 mg/mL, respectively. The time-kill assay results showed that AgNP-K displayed bacteriostatic activity against MRSA. AgNP-K exhibited better antibacterial activity against MRSA when compared to commercial AgNPs or kaempferol alone. - Some of the metrics are blocked by yourconsent settings
Publication The effectiveness of miswak (Salvadora persica L. and Azadirachta indica A. Juss.) practices in reducing plaque and gingivitis among adults: A systematic review and meta-analysis(Elsevier, 2022) ;Haslinda Ramli ;Khairun Nain Nor Aripin ;Shahida Mohd Said ;Rohazila Mohamad HanafiahTuti Ningseh Mohd DomEthnopharmacological relevance: Salvadora persica L. and Azadirachtaindica A.Juss. are listed within the most common sources of miswak or chewing stick that widely used among Western Asia and Muslim populations worldwide. Miswak use in conjunction with toothbrush (adjunctive) has become apparent among the adults. Furthermore, miswak has been reported to have mechanical and pharmacological activities, and benefits to the oral health, by many studies. Aim of the study: To assess the effectiveness of miswak in maintaining periodontal health among adults. Materials and methods: We searched for randomised controlled trials (RCTs) investigating the effect of miswak published in PubMed, EBSCOHOST (Dentistry & Oral Sciences), SCOPUS, and Cochrane Database for Systematic Review (CDSR) from inception to May 08, 2022. The primary outcomes of interest were changes in the periodontal health measured with plaque and gingivitis scores as well as subgingival bacteria load. The quality of evidence was assessed using the Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) approach while the estimates of effect were pooled using a random-effects model. Results: Ten eligible articles were identified, of which 9 could be analysed quantitatively. The remaining report was included as part of the qualitative analysis. The meta-analysis showed that miswak was comparable with the toothbrush in reducing the mean plaque score (p= 0.08, SMD: 0.39, and 95% CI: −0.05 to 0.83) and mean gingivitis score (p= 0.37, SMD: 0.13, and 95% CI: −0.16 to 0.43). Even higher certainty of evidence for the effect of miswak on mean plaque reduction on labial surface of anterior teeth. However, the adjunctive effect of miswak was significantly more superior for reducing plaque (p= 0.01, SMD: 0.68, and 95% CI: 0.14 to 1.22) and gingivitis score (p= 0.04, SMD: 0.66, and 95% CI: 0.03 to 1.29). Conclusions: Miswak effectively reduced plaque and gingivitis scores to a level comparable to toothbrush when used exclusively. Adjunctive miswak use was particularly effective in improving periodontal health. However, the included studies inadequately reported on the method of toothbrushing using miswak and the frequency of miswak use. Therefore, further clinical studies are recommended to explore on the advantages and proper method of miswak practice for optima outcome and safety. - Some of the metrics are blocked by yourconsent settings
Publication Green synthesis, characterisation and antibacterial activities of Strobilanthes crispus-mediated silver nanoparticles (SC-AGNPS) against selected bacteria(Informa UK Limited, 2023-10-17) ;Rohazila Mohamad Hanafiah ;Siti Aisyah Abd Ghafar ;Vuanghao Lim ;Siti Nor Asma Musa ;Fahmi YakopArif Haikal Hairil AnuarThis study aims to characterize and determine the antibacterial activities of synthesized Strobilanthescrispus-mediated AgNPs (SC-AgNPs) against Streptococcus mutans, Escherichia coli and Pseudomonas aer-uginosa. S. crispus water extract acts as a reducing and capping agent in the synthesis of AgNPs. Thesynthesized AgNPs were characterized by using UV-Vis spectrophotometer, dynamic light scattering(DLS), field emission scanning electron microscope (FESEM), X-ray diffractometer (XRD) and Fouriertransform infra-red (FTIR). FESEM images showed a rough surface with a spherical shape. The averagesize distribution of 75.25 nm with a polydispersity index (PDI) of 0.373. XRD analysis matched the face-centred cubic structure of silver. FTIR analysis revealed a shifted peak from 1404.99 to 1345.00 cm−1.MIC and MBC values of SC-AgNPs were 1.25 mg/mL and 2.5 mg/mL against E. coli, P. aeruginosa andS. mutans, respectively. Time-kill assay showed that SC-AgNPs significantly reduced bacterial growth ascompared to non-treated bacteria. Morphologies of bacteria treated with SC-AgNPs were shrunk, lysed,irregular and smaller as compared to control. SC-AgNPs significantly disrupted the gene expression ofeae A, gtf B and Pel A (p < 0.05). This study indicated that the synthesized SC-AgNPs were stable withenhanced antibacterial activities. - Some of the metrics are blocked by yourconsent settings
Publication Identification of Bacteria Sample Plaque from Periodontal Disease Among Diabetic Patients(JK Welfare & Pharmascope Foundation, 2021) ;Shahida Mohd-Said ;Nur Fatimah Zaharah Salehuddin ;Siti Nursyuhada Jafar ;Haslinda Ramli ;Tuti Ningseh Mohd-DomRohazila Mohamad HanafiahMolecular identiβication of bacteria are very important to develop databasegene bank in university. Databases of genes are very important to anno-tate a collection of nucleotide sequence and protein translation of oral bac-teria. This research was conducted to identify bacteria that present in theplaque sample among diabetic patients collected from the Department ofPeriodontology and Community Oral health Universiti Sains Islam Malaysia(USIM). A total of bacteria was successfully isolated on Blood Agar from peri-odontal patients’ plaques. Colony Forming Unit (CFU/mL) was calculated toestimate the number of viable bacteria in a sample after performed serialdilution. Gram stained was done for all isolated samples to differentiatebetween Gram positive and negative organisms. About 2.07 x 107CFU/mLof bacteria was successfully collected from periodontal plaques on diabetespatient. From the numbers, only 17 were selected to identify the species.Seven from the samples were Gram-positive bacteria, meanwhile ten sam-ples were identiβied as Gram-negative bacteria. PCR products were isolatedfrom the samples by using polymerase chain reaction analysis.Streptococ-cus mutans,Streptococcus gordonii,Streptococcus oralis,Streptococcus sangui-nis,Streptococcus agalactiae,Streptococcus lutetiensis,Streptococcus downei,,Aggregatibacter actinomycetemcomitans, Klebsiella pneumoniae,Enterobacterclocae,Salmonella enterica subs enterica,Enterobacter ludwigii, Enterobactermori,Enterobacter roggen kampii,Enterobacter cloacae,Enterobacter tabaciandKlebsiella pneumoniaewere identiβied from all samples via DNA sequenc-ing. - Some of the metrics are blocked by yourconsent settings
Publication Identification Of The Predominant Oral Microbiome In Pericoronitis(Journal of International Dental and Medical Resear, 2019) ;Ainaa Liyana Azemi ;Siti Noor Adnalizawati Adnan ;Rohazila Mohamad Hanafiah ;John Chong Keat Hon ;Ahmad Dzulfikar Bin SamsudinAzmiza Syawani JasniAccumulation of mixed oral microflora appears to be one of the contributing factors to pericoronitis, an inflammation of the oral soft tissues surrounding the crown of a partially erupted, or impacted mandibular third molars.This study was aimed to identify the predominant infectious bacteria related to pericoronitis and their coexistence with other bacterial species at the infection site.Plaque from pericoronal pockets of lower wisdom teeth of 25 patients that have been diagnosed with pericoronitis were sampled and subjected to a standard microbiological procedure for identification of bacterial species including cultivation on enriched agar plates, biochemical profiling and 16s rRNA PCR analysis. A total of 97 microorganisms were isolated and identified from the cultured samples and 94.73% were Gram-positive bacteria; with the highest incidence of Streptococcus gordonii, Streptococcus mitis, and Streptococcus anginosus. This study also revealed that facultative anaerobes were the predominant group causing pericoronitis (89%). The high occurence of multi-strain bacteria ranging from facultative anaerobic to aerobic bacteria display the importance of their infection networks in pericoronitis.Knowledge gained from this study increases our understanding on the role of different pathogens in pericoronitis and provides new insight into the clinical management of patients and in the prevention of its recurrence. Keywords:Pericoronitis, Oral bacteria, Mandibular third molar. - Some of the metrics are blocked by yourconsent settings
Publication Inhibition Mechanism Of Silver Nanoparticle-kaempferol Against Methicillin-resistant Staphylococcus Aureus(Elsevier, 2024) ;Nur Farah Atiqah Mohd Pazli ;Siti Aisyah Abd Ghafar ;Ariff Haikal Hairil AnuarRohazila Mohamad HanafiahMethicillin-resistant Staphylococcus aureus (MRSA), a multidrug resistant strain, is known to cause a threat to public health due to its limited therapeutic treatment. Kaempferol (K) is a natural flavonoid that shows antibacterial activities toward MRSA, but its effectiveness is limited due to its low water solubility. However, poorly aqueous soluble drugs displayed better solubility through nano formulation. Hence, kaempferols were incorporated with silver nanoparticles (AgNPs) to enhance their solubility and antibacterial activity. Previous study showed that AgNPs incorporated with kaempferol (AgNPs-K) exhibited antibacterial activity against MRSA. However, the knowledge regarding the mechanism of action AgNPs-K against MRSA is still limited. The objective of the study is to unravel the inhibition mechanism of silver nanoparticles-kaempferol (AgNPs-K) on treated MRSA. The scanning electron microscopy (SEM) result showed significant difference in morphology between treated and non-treated MRSA which suggest the effectiveness of the AgNPs-K. Non-treated MRSA has an oval shape while MRSA treated with AgNPs-K showed a disrupted cell wall with contents leakage. The transcriptomic profile analysis by Next Generation Sequencing (NGS) showed that various genes and pathways related to biofilm, virulent activity and glycolysis pathway are differently expressed, with 581 genes were downregulated and 641 were upregulated. The affected genes of icab, clfa and eno which involved in biofilm, clumping factor A (virulent) and glycolysis pathway were validated by RT-PCR technique. The results were consistent with the NGS outcome. In conclusion, AgNPs-K possesses antibacterial activity against MRSA and its mechanism of action are reflected in the gene expression of biofilm pathway, virulent and glycolysis activity. Therefore, AgNPs-K can be suggested as a potential alternative to combat MRSA infection. - Some of the metrics are blocked by yourconsent settings
Publication Inhibition Of Streptococcus Mutans Adherence And Biofilm Formation Activities From Melastoma Malabathricum Subfraction(Innovare Academic Sciences, 2018) ;Rohazila Mohamad Hanafiah ;Siti Aisyah Abd Ghafar ;Wan Ahmad Yaacob ;Wan Syaidatul AqmaNazlina IbrahimObjectives: The objectives of the study were to determine antibacterial, anti-adherence, and antibiofilm ctivities of Melastoma malabathricum stem bark acetone extract (MMSBAE) subfraction against Streptococcus mutans. Methods: Fraction 9 (F9) from MMSBAE was subfractionated by thin-layer chromatography (TLC) and analyzed for antibacterial activity against S. mutans by TLC-bioautography. Subfraction 12 (SF12) was isolated from F9 followed by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. Results: MIC and MBC values were 10 mg/mL and 160 mg/mL, indicating bacteriostatic property of SF12. Time-kill assay analysis confirmed bacteriostatic property of SF12 against S. mutans. Crystal violet staining and glass surface assays were used to determine anti-adherence and antibiofilm activities. Concentrations produced 50% reduction in anti-adherence and antibiofilm activities were 40 mg/mL and 20 mg/mL, respectively. Scanning electron microscopy was performed to visualize the effect of SF12 on S. mutans biofilm structure. SF12 was found to lyse biofilm formation on treated bacteria indicating powerful anticariogenic potential against S. mutans. Analysis by quantitative real-time polymerase chain reaction revealed SF12 at MIC value downregulated biofilm formation genes such as gbpA, brpA, gtfC, and comDE. Conclusion: SF12 showed bacteriostatic activities against S. mutans by inhibiting adherence and biofilm activities. - Some of the metrics are blocked by yourconsent settings
Publication Level Of Salivary Microorganisms After Consumption Of Malaysian Tualang Honey: A Preliminary Study(Ectodermal Dysplasia Group-Turkey, 2019) ;Norzalina Ghazali ;Rohazila Mohamad Hanafiah ;Norazlina Mohammad ;Alida MahyuddinZalina BaharudinThis study was conducted to compare the changes of salivary microorganisms after consumption of tualang honey (TH). A total of 44 USIM dental students (male = 9, female = 35) who fulfilled the inclusion criteria participated in this experimental blinded crossover study. Criteria for subject selection were: subjects with no active caries, no history of antibiotic usage for the past 6 months, no history of antimicrobial mouthwash usage for the past 6 months, no orthodontic appliance worn, and healthy. In phase 1, participants in Group A were not given TH to consume whereas Group B consumed honey. After one month washout period, participants in Group A were given TH to consume and Group B had excluded TH from consumption. The mean differences in the salivary bacterial count (CFU/mL) were analysed using repeated measure ANOVA at p value of 0.05. There was not a significant difference in the salivary bacterial count (CFU/mL) at baseline, Day 3, Day 7 and Day 14 during control phase. However, after consumption of tualang honey, the bacterial count was slightly decreased at Day 7, however, the difference was not statistically significant. The increase in CFU count on Day 3 and Day 14 was also not statistically significant. It can be concluded that two weeks consumption of tualang honey did not give any obvious negative effects on the bacterial count. However, further studies will be required to support these preliminary result. - Some of the metrics are blocked by yourconsent settings
Publication Melastoma Malabathricum Stem Bark Acetone Extract As An Anti-bacterial Agent Against Streptococcus Mutans(Pharmascope Foundation, 2020) ;Rohazila Mohamad Hanafiah ;Siti Aisyah Abd Ghafar ;Nur Syahirah Salehuddin ;Wan Syaidatul AqmaNazlina IbrahimMelastoma malabathricum, also known as 'senduduk' in Malaysia, has been used as traditional medicine for diseases such as toothache, dysentery, haemorrhoids and stomachache. Therefore, the objective of this study is to investigate the biological activity of Melastoma malabathricum stem bark extracts (MMSBE) towards Streptococcus mutans. This investigation involved a few methods, which at first is the determination of minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC). Next is by analyzing the time-kill curve, anti-biofilm activity, scanning electron and transmission electron microscopic analyses. Later, next-generation sequencing (NGS) was done to determine differential regulation genes of treated and non-treated S. mutans. Lastly, confirmation of differential regulation genes was done by RT-PCR analysis. As for the results, M. malabathricum stem bark acetone extract (MMSBAE) showed the greatest inhibition concentrations towards S. mutans, followed by M. malabathricum methanol extract (MMSBME). Values of MIC and MBC (MMSBAE) were 1.25 mg/mL and 5 mg/mL. Meanwhile, MIC and MBC values of MMSBME were 5 mg/mL and 40 mg/mL. MMSBAE was chosen to further analyze its anti-bacterial activity against S. mutans. Time kill curve analysis found that MMSBAE possessed bacteriostatic properties against S. mutans. Besides, SEM and TEM analyses revealed that there were some changes to S. mutans cell morphology after treated with MMSBAE while Next gene sequencing analysis revealed significant (p<0.05) gene expression with multiple targets by MMSBAE, which caused inhibition of S. mutans biofilm formation activity. - Some of the metrics are blocked by yourconsent settings
Publication Silver nanoparticles Clinacanthus Nutans leaves extract induced apoptosis towards oral squamous cell carcinoma cell lines(Taylor & Francis Ltd, 2018) ;Fahmi Yakop ;Siti Aisyah Abd Ghafar ;Yoke Keong Yong ;Latifah Saiful Yazan ;Rohazila Mohamad Hanafiah ;Vuanghao LimZolkapli EshakPurpose: The purpose of this study was to investigate apoptotic activity of silver nanoparticle Clinacanthus nutans (AgNps-CN) towards HSC-4 cell lines (oral squamous cell carcinoma cell lines). Methods: Methods involved were MTT assay (cytotoxic activity), morphological cells analysis, flow cytometry and cell cycle analysis and western blot. Results: MTT assay revealed IC50 concentration was 1.61 mu g/mL, 3T3-L1 cell lines were used to determine whether AgNps-CN is cytotoxic to normal cells. At the highest concentration (3 mu g/mL), no cytotoxic activity has been observed. Flow cytometry assay revealed AgNps-CN caused apoptosis effects towards HSC-4 cell lines with significant changes were observed at G1 phase when compared with untreated cells. Morphological cells analysis revealed that most of the cells exhibit apoptosis characteristics rather than necrosis. Protein study revealed that ratio of Bax/Bcl-2 increased mainly due to down-regulation of Bcl-2 expression. Conclusion: AgNps-CN have shown potential in inhibiting HSC-4 cell lines. IC50 was low compared to few studies involving biosynthesized of silver nanoparticles. Apoptosis effects were shown towards HSC-4 cell lines by the increased in Bax/Bcl-2 protein ratio. Further study such as PCR or in vivo studies are required. - Some of the metrics are blocked by yourconsent settings
Publication A simplified system for simulation of Streptococcus mutans biofilm on healthy extracted human tooth as dental plaque model(Open Science Publishers LLP Inc., 2019) ;Nadia Halib ;Nor Zaihana Abdul Rahman ;Rohazila Mohamad Hanafiah ;Nabihah Roslan ;Nurhani Jauhar ;Faculty of DentistryUniversiti Sains Islam Malaysia (USIM)The understanding and capability to simulate reproducible in vitro plaque model are important where this artificial plaque can be utilized as a tool in assessing the efficacy of tooth cleaning products. Previous studies were done on synthetic surfaces such as glass beads under a very strict, controlled environment. Thus, the aim of this study was to establish a simplified system to enable the formation of single species biofilm on the extracted natural tooth as plaque model. Healthy extracted natural tooth was incubated in broth mixture of Streptococcus mutans, containing 5% sucrose and stimulated saliva in a simplified system. The incubation was set at room temperature for 24, 48, and 72 hours. After 24 hours, the sticky deposit was formed on the surface of the tooth. The area of biofilm, stained with disclosing agent, expended from 19.05 ± 1.41 mm2 (24 hours) to 23.25 ± 0.35 mm2 (48 hours). However, no significant expansion of the plaque area was observed on 72 hours (23.83 ± 0.52 mm2). The biofilm was then observed under Infinite Focus Microscope (IFM) to measure the biofilm thickness. The preliminary attempt showed that plaque appeared as dark deposit with a certain thickness. Therefore, this simplified system is a reliable method for producing artificial plaque. - Some of the metrics are blocked by yourconsent settings
Publication Transcriptomic Profile Analysis of Streptococcus mutans Response to Acmella paniculata Flower Extracts(Hindawi, 2022) ;Siti Aisyah Abd Ghafar ;Nur Syahirah Salehuddin ;Nor Zaihana Abdul Rahman ;Nadia HalibRohazila Mohamad HanafiahBackground. Acmella paniculata has been used as a traditional medicine to treat oral health diseases such as dental caries and periodontitis. Streptococcus mutans is a common bacterium that initiates dental caries at an early stage. Aim. The aim of this study was to determine the mode of action of A. paniculata (extracts) against S. mutans growth. Methods. Time-kill assay has been done to investigate the rate of kill and effectiveness of Acmella paniculata (AP) extracts against S. mutans growth. Phytochemical analysis was done to identify major compounds in AP extracts using gas chromatography mass spectrometry (GCMS). Scanning and transmission electron microscopy (SEM and TEM) have been done to observe the morphological changes of treated bacteria. Transcriptomic profile analysis has been done using Next Gene Sequencing. Results. AP flower n-hexane (APFH) and AP flower dichloromethane (APFD) extracts acted as bactericidal agents after killing >3 log10 cfu/mL of S. mutans after 24 hours. Oleic and hexadecenoic acids were found to be the major compounds in APFD and APFH extracts, respectively. Photomicrographs from SEM and TEM of treated S. mutans show that the bacterial cell wall has been lysed and the cytoplasm content was decreased. Pathway analysis revealed that the APFD extract significantly affected biosynthesis peptidoglycan, gene expression, RNA processing, and macromolecule metabolism processes in S. mutans. Conclusion. Data analysis revealed that multiple mechanisms of action were involved in antibacterial activity of A. paniculata extracts toward S. mutans.