ORIGINAL RESEARCH

Antifungal Activity of Aloe Vera Leaf and Gel Extracts

Against Candida albicans: An In Vitro Study
M Shilpa1, Vinaya Bhat2, A Veena Shetty3, Mora SR Reddy4, Prashant Punde5

A b s t r ac t​
Aim: The present study was conducted with an aim to assess the antimicrobial activity of ethanolic extracts of aloe vera leaf and gel against
Candida albicans in vitro.
Materials and methods: Fresh leaves were collected from the aloe vera plants naturally grown in Coorg. Aloe vera leaf as well as gel was separated,
extracted with 95% ethanol in rotary shaker at constant temperature for 3 days, evaporated in a heating mantle, and stored in screw cap test
tubes at 4°C for further analysis. Antifungal activity of aloe vera leaf and gel extracts against C. albicans was assessed by well diffusion method.
Further, gel extracts of aloe vera at different dilutions (500, 400, 300, and 200 μL) were prepared and the turbidity was analyzed.
Results: Results showed that the ethanolic extract of aloe vera leaf did not show antifungal activity against C. albicans. A maximum of 99.33%
antifungal activity was shown by 400 μL of aloe vera gel extract. The minimum inhibitory concentration of aloe vera gel extract was 200 μL
(98.2% inhibition).
Conclusion: The ethanolic extract of aloe vera gel showed considerable antifungal activity against C. albicans.
Clinical significance: Modalities targeted on the use of aloe vera against C. albicans can prove to be more beneficial and consistent compared
to conventional antifungals for preventive and/or therapeutic purposes against a variety of oral fungal diseases.
Keywords: Aloe vera, Antifungal, Candida albicans, Natural.
World Journal of Dentistry (2020): 10.5005/jp-journals-10015-1701

I n t r o d u c t i o n​
Natural products are important resources in traditional medicine
and have been long used for prevention and treatment of many
diseases.1 All over the globe, many plants have been exploited
for their medicinal value. Mukherjee and Wahile reported about
the considered opinion of World Health Organization, which
states that, “80% of the world’s population are dependent on
ancestral medicines for their haleness”. For the healthcare of
the remaining 20% population mainly residing in developed
countries, therapeutic product of plants plays an important
role. 2
Aloe vera (Sanskrit-Ghritakumari, Kumara; Hindi-Guarpatha,
Ghikanvar) a herb, commonly referred to as the “medicinal plant”,
is known for its wide range of therapeutic properties. The botanical
name of aloe vera is Aloe barbadensis Miller and it belongs to
lily family. Aloe products are very popular in the market and are
widely used in skin care, cosmetics, medical, healthcare, and food
industry.3,4
Aloe vera is made up of many complex ingredients including
polysaccharides, glycoproteins, phenolic compounds, salicylic acid,
lignin, hormones, amino acids, vitamins, saponins, and enzymes,
which give aloe vera its many beneficial properties including anti-
inflammatory, antibacterial, antioxidant, immune-boosting, and
hypoglycemic properties.3,5
Even though several effective antifungal agents are available
for oral candida infections, the failure is not uncommon because
isolates of C. albicans may exhibit resistance to the drug during
therapy.6
Hence, the present study was conducted with an aim to assess
the antimicrobial activity of ethanolic extracts of aloe vera leaf and
gel against C. albicans in vitro.
1
Department of Public Health Dentistry, AB Shetty Memorial Institute
of Dental Sciences, NITTE (Deemed to be University), Mangaluru,
Karnataka, India
2
Department of Prosthodontics, AB Shetty Memorial Institute of Dental
Sciences, NITTE (Deemed to be University), Mangaluru, Karnataka,
India
3
Department of Microbiology, KS Hegde Medical Academy (KSHEMA),
NITTE (Deemed to be University), Mangaluru, Karnataka, India
4
Department of Orthodontics, Kalinga Institute of Dental Sciences, KIIT
Deemed to be University, Bhubaneswar, Odisha, India
5
Department of Oral and Maxillofacial Surgery, School of Dental
Sciences, Krishna Institute of Medical Sciences (Deemed to be
University), Karad, Maharashtra, India
Corresponding Author: Vinaya Bhat, Department of Prosthodontics,
AB Shetty Memorial Institute of Dental Sciences, NITTE (Deemed to
be University), Mangaluru, Karnataka, India, Phone: +91 9481921180,
e-mail: drvinayabhat@gmail.com
How to cite this article: Shilpa M, Bhat V, Shetty AV, et al. Antifungal
Activity of Aloe Vera Leaf and Gel Extracts Against Candida albicans: An
In Vitro Study. World J Dent 2020;11(1):36–40.
Source of support: Nil
Conflict of interest: None
M at e r ia l s and M e t h o d s​
The study protocol was approved by the Institutional Review Board
of Coorg Institute of Dental Sciences, Virajpet.
Preparation of the Ethanolic Extract of Aloe Vera Gel
Fresh leaves weighing 1 kg were collected from the aloe vera plants
(A. barbadensis Miller, belonging to Liliaceae family) naturally grown

© The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.
org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and non-commercial reproduction in any medium, provided you give appropriate credit to
the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain
Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
 Aloe Vera Leaf and Gel Extracts against Candida albicans
in Coorg. After washing the leaves with tap water and then again
rinsing with distilled water, the leaves were dissected with a sterile
knife into longitudinal sections and gel was scooped out with a
sterile sharp spatula, care being taken to avoid the mixture of the
fibers into the gel.
Aloe vera gel was macerated in an electric blender to obtain
a fine paste (Fig. 1), by adding few drops of 95% ethanol. The
macerated gel was equally distributed in conical flasks after
which 150 mL of 95% alcohol was added to all the conical flasks.
Subsequently, the flasks were kept in rotary shaker for 3 days.
After 3 days, the macerated gel was filtered using Whatman
filter paper no. 1. It was then evaporated in a heating mantle and
stored in a screw cap test tube at 4°C.
Preparation of the Ethanolic Extract of Aloe Vera Leaf
The leaves which were separated from the gel were washed in
distilled water, cut into small pieces with a sterile knife, and kept
for drying. After 24 hours, they were pressed with filter paper to
remove the moisture content and residual gel of the leaves. Then the
leaves were ground into a fine paste using few drops of 95% ethanol.
The ground leaf paste was mixed with 600 mL of 95% ethanol
along with constant stirring on a hot water bath at around 45 to
50°C. The mixture was then kept in a magnetic stirrer at a constant
temperature of about 38°C for 3 days (Fig. 2) after which it was
filtered using Whatman filter paper no. 1. The solution obtained
was collected and reduced in the heating mantle to obtain thick
dark brownish viscous syrup. This crude ethanolic extract of aloe
vera leaf was stored at 4°C for further analysis.
Assessing Antifungal Activity of Aloe Vera Leaf and
Gel Extracts by Well Diffusion Method
Preparation of C. albicans Culture
Pure culture of C. albicans ATCC 90028 strain was cultured in
Sabouraud dextrose agar (Himedia laboratories, Mumbai, India)
and maintained at 37°C for 24 hours. The 24-hour culture was
adjusted to match the 0.5 McFarland turbidity standard to obtain
final inoculum of 2 × 104 cfu/mL.
Sterile Sabouraud dextrose agar plates were prepared, one
each for the leaf and gel of aloe vera, respectively. After swabbing
each plate with the suspension of C. albicans, four wells of 8 mm
were prepared using a sterile metallic template in both the plates.
Fig. 1: Macerated aloe vera gel
World Journal of Dentistry, Volume 11 Issue 1 (January–February 2020)
Inoculation of the Test media
The leaf extract 500 mg was diluted in 1 mL of dimethyl sulfoxide
(DMSO), of which 500 μL of the extract was dispensed into each
of the three prepared wells in the first petri plate. Also, 2 g of the
gel extract was diluted in 2 mL of DMSO, of which 500 μL of the
extract was dispensed into each of the three wells in the second
petri plate. The fourth wells in both the plates served as positive
control (Fluconazole, 25 μg) and were incubated for 24 hours at
37°C. Hence, both the tests were conducted in triplicates.
After incubation, the zones of inhibition (i.e., locations where no
growths of Candida were present) were examined around the wells
that contained the test samples. There was no zone of inhibition
around the well that contained aloe vera leaf extract samples (Fig. 3).
There was a clear zone of inhibition around the well containing aloe
vera gel extract samples (Fig. 4). Therefore, microbroth dilution
method was used to examine the extent of antifungal activity of
aloe vera gel extract against C. albicans.
Assessing Antifungal Activity of Aloe Vera Gel Extract
by Microbroth Dilution Method
Preparation of C. albicans Culture
Pure culture of C. albicans ATCC90028 strain was inoculated in yeast
nitrogen base (YNB) and incubated overnight at 37°C for 24 hours. It
was then centrifuged at 3,000 rpm for 5 minutes at three successive
intervals and the supernatant was discarded each time.
The pellet was then resuspended in YNB and the turbidity
was adjusted to match the 0.5 McFarland turbidity standard. The
McFarland adjusted culture was diluted in 14 mL of Roswell Park
Memorial Institute 1640 Medium (RPMI-1640) in a 1:50 ratio.
Preparation of the Test Samples
In a screw cap test tube, 4 g of the aloe vera gel extract was weighed
and diluted with 4 mL of DMSO. Gel extracts of different dilutions
(500, 400, 300, and 200 μL) were prepared.
Assessing the Antifungal Activity
Subsequently, 500 μL of the RPMI + culture was collected in cuvettes
to which gel extracts of different dilutions (500, 400, 300, and 200 μL)
were added. The cultures were then incubated for 24 hours. After
24 hours, the biochemical analyzer was used to check the turbidity.
Descriptive statistics was used to summarize the results (Table 1).
Fig. 2: Ground aloe vera leaf paste – ethanol mixture in the magnetic
stirrer
37
 Aloe Vera Leaf and Gel Extracts against Candida albicans

Fig. 3: Microscopic view depicting no zone of inhibition for 500 μL of Fig. 4: Clear zone of inhibition observed for 500 μL of aloe vera gel
aloe vera leaf extract against Candida albicans extract against Candida albicans

Table 1: Optical densities of the Aloe vera gel extract samples R e s u lts​
S. no. Test sample Dilutions (μL) Optical density Antifungal Activity by Well Diffusion Method (Aloe
1 Aloe vera gel extract + 500 400 Vera Leaf and Gel Extracts) and Microbroth Dilution
RPMI + culture
Method (Aloe Vera Gel Extract)
2 Aloe vera gel extract + 400 460
There was no zone of inhibition around the well that contained
RPMI + culture
aloe vera leaf extract samples (Fig. 3). There was a clear zone of
3 Aloe vera gel extract + 300 361.5 inhibition around the well containing aloe vera gel extract samples
RPMI + culture (Fig. 4). As shown in Table 2 and Figure 5, the maximum inhibition
4 Aloe vera gel extract + 200 364.5 of 99.33% was shown by 400 μL of the gel extract, while 94.2% of
RPMI + culture inhibition was shown by 500 μL and 300 μL of the gel extract. From
5 Aloe vera gel extract + 500 474 Figure 6 it is clear that, per the National Committee for Clinical
RPMI Laboratory Standards (NCLSS) guidelines for microbroth dilution,
6 Aloe vera gel extract + 400 451 the minimum inhibitory concentration of aloe vera gel extract was
RPMI 200 μL (98.2% inhibition).
7 Aloe vera gel extract + 300 439 Growth (%) and inhibition (%) was derived by applying the
RPMI following formula for optical density (OD) of test samples:
8 Aloe vera gel extract + 200 340
RPMI (OD of broth + drug - OD of drug)
Growth % = × 100
(All values are mean of triplicates) OD of broth

i.e., growth% =
Table 2: The inhibition percentage obtained after substituting the above (OD of RPMI + gel + culture) - (OD of RPMI + gel)
× 100
formula with the values obtained for test samples as shown in Table 1 OD of RPMI + culture
S. no. Test sample Dilutions (μL) Growth (%) Inhibition (%)
Hence, inhibition% = 100 – growth%
1 Aloe vera gel 500 5.8 94.2
extract + RPMI +
culture D i s c u s s i o n​
2 Aloe vera gel 400 0.67 99.33 Among the Candida species, C. albicans frequently has been reported
extract + RPMI + to cause oral infections in immunocompromised individuals due
culture to suppression of local and systemic defense mechanisms.7,8 They
3 Aloe vera gel 300 5.8 94.2 can also persist and contaminate the endodontically treated teeth
extract + RPMI + resulting in the failure of treatment.9 The use of natural products as
culture alternative agents for the control of fungal diseases is considered
4 Aloe vera gel 200 1.8 98.2 as an interesting alternative to synthetic fungicides.6
extract + RPMI + Antifungal activity was recorded by the aloe vera gel extract
culture only and not by the leaf extract, against C. albicans, in the present

38 World Journal of Dentistry, Volume 11 Issue 1 (January–February 2020)

 Aloe Vera Leaf and Gel Extracts against Candida albicans

Fig. 5: Growth (%) and inhibition (%) of different dilutions of aloe vera gel extract

Fig. 6: Inhibition (%) of different dilutions of aloe vera gel extract

study. This is similar to the study by Khaing, where the aloe vera leaf
extract did not display any antifungal activity against C. albicans.10
In the study by Agarry et al., the growth of C. albicans was inhibited
by aloe vera leaf but not by the gel.11
Regarding the antifungal activity of various dilutions of aloe
vera gel extract against C. albicans, a maximum inhibition of 99.33%
was shown by 400 μL of the aloe vera gel extract, while 94.2% of
inhibition was shown by 500 and 300 μL of the gel extract. In the
study by Tamilarasi et al., 150 μL of the ethanolic extract of aloe
vera gel showed the strongest inhibitory activity against C. albicans
(79%).12
The degree of inhibition varied depending upon the dilutions
of the gel extracts in the present study. In the study by Devi et al.
and Renisheya et al., antifungal activity of the aloe vera gel DMSO
extracts also varied according to the concentration, with the highest
antifungal activity shown by 400 μg/mL, followed by 200 and
100 μg/mL.13,14
The minimum inhibition concentration that was obtained in
the present study was 200 mg/mL (98.2%). This is in contrast to the
study by Stanley et al., which revealed that ethanol extract of aloe
vera gel was susceptible to C. albicans, and the minimum inhibitory
concentration of ethanolic extract of aloe vera gel on C. albicans
was 0.50 mg/mL.15
It is evident from the present investigation that ethanolic
extract of aloe vera gel has antifungal effects against C. albicans,
which lends more weight to the general acceptability of herbal
extracts for therapeutic purposes.
Various studies have been done to assess the antimicrobial
activity of aloe vera on C. albicans. Aloe vera was observed to
have antifungal activity against oral thrush.16 It has also been
reported that a processed aloe vera gel preparation inhibited the
growth of C. albicans.17 Aloe vera leaf extracts have been observed
to inhibit the germ tube formation and hence the growth of
C. albicans. The purified aloe protein has been found to exhibit
potent antifungal activity against Candida paraprilosis, Candida
krusei, and C. albicans.18
Previous reports suggest that aloe vera could inhibit infectious
diseases by stimulating the host defense mechanism, especially
the phagocytic and killing activities of macrophages. Evidence
supports the usage of aloe vera as an antimicrobial agent

World Journal of Dentistry, Volume 11 Issue 1 (January–February 2020) 39

 Aloe Vera Leaf and Gel Extracts against Candida albicans
and promotes its use in research, especially in toxicology and
pharmacology.12
The antimicrobial effect of a dentifrice containing aloe vera has
been demonstrated in an in vitro study, in which this phytotherapic
agent inhibited the growth of diverse oral microorganisms, such as
Streptococcus mutans, Streptococcus sanguis, Actinomyces viscosus,
and C. albicans.19 The results of the present study lend support to
the using of the aloe vera gel extracts as home remedies or adding
to dentifrices, mouthwashes, and varnishes which may create an
oral environment which is unfavorable for C. albicans.
It has been reported that aloe vera gel contains glycoprotein
with cell proliferating–promoting activity and that it improves
wound healing by increasing angiogenesis resulting in increased
oxygenation. This is particularly beneficial in patients with sore
gums and teeth as well as denture stomatitis.5,20
With the limitation of the study being assessment of antifungal
activity using a single extraction method, i.e., ethanolic extract
of aloe vera leaf and gel, further studies should be carried out
using various extraction procedures along with clinical testing to
ascertain the comparative antifungal activity of aloe vera leaf and
gel against C. albicans.
C o n c lu s i o n​
From the study results, it was concluded that the ethanolic extract
of aloe vera gel showed considerable antifungal activity against
C. albicans. There was varying antifungal activity according to
varying dilutions of aloe vera gel extract. The minimum inhibitory
concentration of aloe vera gel extract was 200 μL (98.2% inhibition),
whereas the maximum of 99.33% antifungal activity was shown by
400 μL of aloe vera gel extract.
References
1. Farahnejad Z, Ghazanfari T, Yaraee R. Immunomodulatory effects
of Aloe vera and its fractions on response of macrophages against
Candida albicans. Immunopharmacol Immunotoxicol 2011;33(4):
676–681. DOI: 10.3109/08923973.2011.560158.
2. Raksha B, Pooja S, Babu S. Bioactive compounds and medicinal
properties of Aloe vera L.: an update. J Plant Sci 2014;2(3):102–107.
3. Sharma A, Gautam S. An overview on medicinal properties of
Aloe vera: antibacterial & antifungal aspects. Int J Pharm Bio Sci
2013;4(3):694–705.
40 World Journal of Dentistry, Volume 11 Issue 1 (January–February 2020)
4. Gupta VK, Malhotra S. Pharmacological attribute of Aloe vera:
revalidation through experimental and clinical studies. Ayu
2012;33(2):193–196. DOI: 10.4103/0974-8520.105237.
5. Shireen F, Manipal S, Prabu D. Anti-fungal activity of Aloe vera: in
vitro study. SRM J Res Dent Sci 2015;6(2):92–95. DOI: 10.4103/0976-
433X.155464.
6. Doddanna SJ, Patel S, Sundarrao MA, et al. Antimicrobial activity of
plant extracts on Candida albicans: an in vitro study. Indian J Dent
Res 2013;24(4):401–405. DOI: 10.4103/0970-9290.118358.
7. Hegde V, Rodrigues SJ, Saldanha S. Redefining the treatment
modalities for denture stomatitis- a review. Int J Res Dent 2014;4(2):
8–27.
8. Pereira-Cenci T, Del Bel Cury AA, Crielaard W, et al. Development of
candida-associated denture stomatitis: new insights. J Appl Oral Sci
2008;16(2):86–94. DOI: 10.1590/s1678-77572008000200002.
9. Shah N, Madhu KS, Sreenivasa Murthy BV, et al. Identification of
presence of Candida albicans in primary root canal infections: an in
vitro study. Endodontology 2016;28(2):109–113. DOI: 10.4103/0970-
7212.195440.
10. Khaing TA. Evaluation of the antifungal and antioxidant activities of
the leaf extract of Aloe vera (Aloe barbadensis miller). Proc World
Acad Sci, Eng Tech 2011;75:610–612.
11. Agarry OO, Olaleye MT, Bello-Michael CO. Comparative antimicrobial
activities of aloe vera gel and leaf. Afr J Biotechnol 2005;4:1411–1414.
12. Tamilarasi L, Ponnulakshmi R, Ezhilarasi BS. Comparative study on
the antifungal activity of Aloe vera leaf and gel in different extracts.
Int J Pharm Bio Sci 2014;5(2):B895–B901.
13. Devi DL, Srinivas B, Rao BN. An evaluation of antimicrobial activity of
Aloe barbadensis miller (Aloe vera) gel extract. JPBMS 2012;21(03):1–4.
14. Renisheya T, Johnson M, Nancy BS, et al. Anti-bacterial and antifungal
activity of aloe veragel extract. IJBAR 2012;03(03):184–187.
15. Stanley MC, Ifeanyi OE, Eziokwu OG. Antimicrobial effects of aloe
vera on some human pathogens. Int J Curr Microbiol App Sci
2014;3(3):1022–1028.
16. Bhat G, Kudva P, Dodwad V. Aloe vera: Nature’s soothing healer to
periodontal disease. J Indian Soc Periodontal 2011;15(3):205–209.
DOI: 10.4103/0972-124X.85661.
17. Sundarkar P, Govindwar R, Nyamati SB, et al. Use of aloe vera in
dentistry. J Indian Acad Oral Med Radiol 2011;23(3):S389–S391.
18. Tayal E, Sardana D, Indushekar KR, et al. Current perspectives on use
of aloe vera in dentistry. Eur J Med Plants 2014;4(12):1408–1419. DOI:
10.9734/EJMP/2014/10843.
19. Agarwal A, Dwivedi N. Aloe vera: magic or myth. SRM J Res Dent Sci
2013;4(3):119–124. DOI: 10.4103/0976-433X.121638.
20. Isadkar YS, Palaskar SJ, Narang B, et al. Aloe vera as denture cleanser.
J Dent Allied Sci 2018;7(1):23–26. DOI: 10.4103/jdas.jdas_44_17.