Alojid, Anis Amiera Muhamad
(2023)
The cytotoxicity and antimicrobial
activity of andrographis paniculata herbal
mouthwash formulation.
Masters thesis, Universiti Sains Malaysia.
Abstract
Oral disease is a global health problem caused by oral pathogens. To prevent
the oral disease, mouthwash is commonly used. However, commercial mouthwash
contained alcohol and other chemicals found to be toxic with unfavourable side effects.
Besides, existence drugs seemed to be ineffective due to excessive use and multidrug
resistance against the oral pathogens. Thus, there is a demand for a safer and effective
alternative treatment in dental product. As the studies on cytotoxicity and antimicrobial
activities of medicinal herb, Andrographis paniculata (AP) against oral pathogens are
limited, this study proposed AP aqueous extract as herbal mouthwash to determine its
effectiveness against selected oral pathogens; Porphyromonas gingivalis,
Streptococcus mutans, S. sobrinus, Staphylococcus aureus, and Actinomyces viscosus.
In this study, antibacterial effect of AP aqueous extract against the tested oral
pathogens was determined by disc diffusion and well diffusion assays, followed by
antibacterial effect of five concentrations of AP herbal mouthwash at 1000, 500, 250,
125 and 62.5 mg/mL using well diffusion and broth microdilution method. Assessment
of toxicity was determined by brine shrimp lethality bioassay and 3-(4,5-
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay onto L929
fibroblast cells while the analysis of heavy metal was analysed using atomic absorption spectrometry (AAS). Stability study was carried out on the final product of AP herbal
mouthwash. Results revealed that AP aqueous extract significantly inhibited the
growth of all oral pathogens (p<0.05) except for S. aureus with inhibition zones
ranging from 2.83 ± 0.83 mm to 14.37 ± 0.83.mm. Similar to AP aqueous extract, AP
herbal mouthwash exhibited significant antibacterial activities onto all tested pathogens except for S. aureus (p<0.05) with inhibition ranging from median (IQR) of
9.50 (3.80) to median (IQR) of 15.00 (3.40). All mouthwash concentrations caused
bacteria inhibition from 10.56% to 261.09% with minimal inhibitory concentration
(MIC) and minimal bactericidal concentration (MBC) at 3.90 mg/mL to 7.81 mg/mL
and 7.81 mg/mL to 62.50 mg/mL respectively. However, there was no MBC detected
for S. aureus at these concentrations. Formulation of mouthwash was non-toxic to
brine shrimp and L929 fibroblast cell line with LC50 value 3255.06 μg/mL and IC50
value 43.55 mg/mL respectively. AP herbal mouthwash was free from heavy metal
contamination and maintained its characteristics and sterility for 12 months storage. In
conclusion, AP herbal mouthwash at 7.81 mg/mL was found to be the best mouthwash
formulation. However, more research works need to be carried out for its clinical use
for further product commercialization.
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