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Article

Study of the Cosmetic Potential Uses of Plants from Mayotte as Skin Care Agents through the Screening of Their Biological Activities

Laboratory of Chemistry of Natural Molecules, Gembloux Agro-Bio Tech, University of Liège, Passage des Déportés, 2, 5030 Gembloux, Belgium
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Nutraceuticals 2022, 2(4), 420-440; https://doi.org/10.3390/nutraceuticals2040031
Submission received: 27 October 2022 / Revised: 15 November 2022 / Accepted: 23 November 2022 / Published: 1 December 2022

Abstract

:
(1) The island of Mayotte, located in the Indian Ocean, possesses remarkable biodiversity. In a previous ethnobotanical study, we already highlighted 69 different plant species used in traditional medicine. Among those, 21 were traditionally employed for skin care by the local populations. The present study aimed to better understand the traditional use of those plants by investigating their in vitro biological activities and, more specifically, their anti-lipoxygenase, antioxidant and anti-tyrosinase properties. (2) These three activities were, respectively, determined by lipoxygenase inhibition, DPPH-reducing potency, and anti-tyrosinase activity assays. (3) Results revealed high biological activities for several plant species, with some of them displaying one strong single activity, while others had at the same time anti-lipoxygenase, antioxidant and anti-tyrosinase effects. (4) Those in vitro biological activities agreed with the traditional use of those plants by the local population. It also highlights the high potential of those species from Mayotte in the development of new cosmetic ingredients for the treatment of many skin affections, such as eczema.

Graphical Abstract

1. Introduction

The island of Mayotte is part of the Comoros archipelago located in the Indian Ocean. One of the particularities of this island is its localization in a geographical area where the climate is appropriate for the development of exceptional biodiversity. Indeed, it has been reported that 25% of the earth’s biodiversity is present in the Indian Ocean area [1]. It was around 1820 that the interest in tropical islands from a botanical point of view began to grow, as De Candolle [2] notably suggested that “…‘Islands’ flora is worth studying due to the peculiar characteristics shown by their vegetation or to the fact that the study areas are clearly defined, thus the studies can be done thoroughly…”. Since then, this affirmation has been confirmed many times in numerous insular areas of the world. However, Mayotte was not one of them, as very little research has been done about this small island [2].
On the other hand, the history of the island of Mayotte and, notably, the successive settlement of people from African Bantu and Arab-Muslim origins [3] explains that the use of traditional medicine is still very common and deeply ingrained in the habits of the inhabitants [4,5,6]. Indeed, as these people have a strong bond to their cultural heritage, traditional cosmetic practices are still used nowadays. The conservation of this knowledge is based on an oral transmission that passes from one generation to another [7,8]. Mayotte has, therefore, much to offer when it comes to ethnobotanical studies [9].
During a previous ethnobotanical field study on the Island of Mayotte, we identified 69 plants used in traditional medicine for the treatment of many different conditions [10]. Among these plants, we noted a large number of uses related to skin care, whether to treat skin problems or for cosmetic purposes [10]. As the skin is the first protective layer of the human body, it withstands all types of stress, either endogenous, such as the effect of aging, or exogenous, such as sun exposition. These stresses are responsible for many skin imperfections and afflictions [11]. It did not take long for humanity to understand that these deleterious effects could be thwarted using what nature has to offer. Therefore, plants have been used to heal but also for aesthetic uses through makeup and perfume for as long as humanity can remember. These practices are deeply linked to man’s evolution [12,13].
In the present work, we focused more particularly on 21 plants traditionally used for skin care by the inhabitants of Mayotte based on our previous ethnobotanical study. Indeed, pharmaceutical industries are continuously searching for new bioactive molecules. The main purpose of the present study was, therefore, to investigate the potential of these plants for use as an ingredient in cosmetics through the study of three targeted biological activities: anti-lipoxygenase activity linked to anti-inflammatory properties, antioxidant activity, and anti-tyrosinase activity related to the treatment of pigmentation issues.
The inflammatory triggers can be divided into infections, tissue injuries, as well as tissue stress and malfunction. In the present work, we will focus on the second and third types of affection since the anti-inflammatory properties are meant to be paired with cosmetic practices. Among the different pathways linked to the appearance of inflammatory reactions, the research for lipoxygenase (LOX)-inhibitory compounds is crucial as LOX is responsible for the synthesis of leukotrienes, which are pro-inflammatory molecules [14]. Concerning antioxidant activity, we focused on the reaction involving DPPH (2,2-Diphenyl-1-picrylhydrazyl), which has previously been described as a good indicator of the antioxidative power [15]. This specific test assay indeed detects the scavenging potency of free radicals through the scavenging activity of the stable DPPH free radical. Finally, the tyrosinase inhibition capacity of plant extracts was used to evaluate their potential to treat skin pigmentation issues. Indeed, tyrosinase is the key enzyme in the biosynthesis of melanin that plays a major protective role against skin photocarcinogenesis, such as melanoma. Melanin is also responsible for phenomenon known as hyperpigmentation and hypopigmentation, often visible in the form of darker or lighter spots, zones or body parts [16]. The search for natural chemical agents capable of modulating pigmentation metabolism is, therefore, of great interest.

2. Materials and Methods

2.1. Reagents

Acetone (≥99%, technical), methanol (≥98.5%, technical), K2HPO4, KH2PO4, pure ethanol, NaOH, and H3BO3 were purchased from VWR chemicals (Leuven, Belgium). DPPH (2,2-Diphenyl-1-picrylhydrazyl), mushroom tyrosinase (EC 1.14.18.1), kojic acid, TROLOX ((±)-6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), linoleic acid, Tween 80, L-DOPA (3,4-dihydroxiphenilalanine) and Glycine max lipoxygenase (EC 1.13.11.12) were purchased from Sigma-Aldrich (Darmstadt, Germany).

2.2. Plant Extracts Preparation

Based on the results of the previous ethnobotanical survey conducted by M. Saive in 2014 on the island of Mayotte [10], 21 different plant species were collected, and a specimen was stored for each sample at the CBNM herbarium in Mayotte [10]. All available organs at the time of the harvest were collected, leading to a total of 89 samples (see Table 1).
All samples were dried at 40 ± 1 °C in a drying oven for 48 h. When the vegetal material allowed it, the samples were powdered using an analytical grinder IKA A11 (Staufen, Germany). The global size of the ligneous parts of the vegetal material was reduced using shears. Once the size allowed it, the samples were then ground using a laboratory hammermill mounted with a 6 mm mesh. The ground samples were kept vacuum-packed at −22 ± 1 °C until extracted. The extraction process was carried out using a Soxhlet apparatus with acetone as a solvent. Acetone is indeed recognized as a solvent of choice for the extraction of bioactive compounds and, notably, antioxidants [17]. Two g of dried and ground samples were weighed and poured into the extraction chamber. Thirty mL of acetone was used [18], and the extraction lasted for 6 h. Once the extraction was completed, the crude extract was evaporated using a rotary evaporator (Heidolph, Laborota 4003, Schwabach, Germany). The dried crude extracts were kept at −22 ± 1 °C until used.

2.3. Lipoxygenase Inhibition Evaluation

The lipoxygenase inhibition capacity of plant extracts obtained by Soxhlet extraction with acetone was assessed by a spectrophotometric method described by Tanoh et al. (2019) with some minor modifications [19].
Lipoxygenase is sensitive to heat; therefore, all the following experimental steps were performed on an ice bath. A 0.1 mg/mL lipoxygenase solution (≥50,000 U/mg) was prepared using distilled water. The 1 mM linoleic acid substrate solution was prepared as follows: 140 mg of linoleic acid, 18 mg of Tween 80, and 100 µL of NaOH 5 mM were mixed and adjusted to 50 mL with distillate water. The enzyme solution, as well as the substrate, were divided into 1 mL aliquots and stored at −22 ± 1 °C until used, with only one thaw-out allowed.
As the expected product for the reaction was 13-HPOD, the optimal pH of the reaction matrix was obtained using a 0.1 M sodium borate buffer corrected to pH 9.5 using 5 M NaOH. To avoid false-positive results due to enzyme inactivation caused by acetone, pure ethyl alcohol was used for sample preparation. Five mL of pure ethanol was added to the dry extracts, which were then submitted to sonication and subsequently filtered using 0.45 µm PTFE syringe filters (Whatman, Puradisc 13, Maidstone, UK). As the color of the crude extracts was an issue, all samples were diluted in the borate buffer from factor 101 to factor 104. As oxygen is a co-substrate for the reaction, the borate solution was oxygenated for 30 min at room temperature prior to the reaction. The control was realized using 0.1 M borate buffer, leading to a non-inhibited reaction, which can be expressed as 0% of inhibition, and the blank was obtained by carrying out the experiment without the enzyme, which causes the absence of 13-HPOD synthesis and can be considered as 100% of inhibition.
The enzymatic reaction was performed by mixing 100 µL of plant sample with 35 µL of lipoxygenase and 800 µL of the oxygenated buffer. For the control, 900 µL of the oxygenated buffer was used. For the blank, 900 µL of oxygenated buffer was used, but no enzyme was added prior to the incubation. The mixtures were left to incubate for 15 min at room temperature and then added into quartz absorption cells along with 35 µL of the substrate solution. The synthesis of linoleic acid hydroperoxide was observed in real time as absorbance was measured every 20 s for 5 min at 234 nm. Each sample/control was done in triplicate. The relative activity was obtained by putting the slopes of the samples against the slopes of the control, according to Equation (1):
%   inhibition   activity =   Control   Slope   Sample   Slope   Control   Slope × 100
where the control slope is the slope of the control (0% inhibition) at the linear part of the reaction, and the sample slope is the slope of the samples at the linear part of the reaction, defined by an R2 higher than 0.9.

2.4. DPPH-Reducing Potency Evaluation

The evaluation of the DPPH-reducing potency of the plant samples was based on a protocol adapted from M. S. Blois (1958) [20]. A 2 × 10−4 M DPPH solution was prepared using technical-grade methanol. Once prepared, this solution was kept at 4 ± 1 °C in the dark and was used within the following 7 days.
The positive control for this test was a 2 × 10−3 M TROLOX solution prepared with methanol. The blank was likewise performed with methanol. For the sample analysis, 5 mL of methanol was added to the dry extracts, which were obtained by Soxhlet extraction with acetone. The crude extract was retrieved by sonication. The solution was then filtered using 0.45 µm PTFE syringe filter (Whatman, Puradisc 13, Maidstone, UK) and submitted to successive dilutions (10 to 10,000 times) in methanol. The analysis went as follows. A 1:1 DPPH:sample/blank/TROLOX mix was performed and left to incubate at room temperature for 10 min. Then the absorbance of the reaction mix was observed using an Ultrospec 9000 UV/Vis spectrophotometer from Biochrom (Cambridge, UK) at 517 nm. Each sample was measured in triplicate. The samples’ relative activity was established by comparing the final absorbance with the absorbance obtained with the blank and with the TROLOX, as seen in Equation (2):
%   reduction   activity = ( 1     Abs   TROLOX     Corrected   value Abs   TROLOX     Abs   BLANK )   ×   100
where Abs TROLOX refers to the absorption at 517 nm of the positive control (100% of activity), and Abs BLANK refers to the absorption at 517 nm of the negative control (0% of activity). The corrected value is obtained by Equation (3):
Corrected   value = Final   abs     Sample   abs
where the final abs is equal to the absorbance (517 nm) of the reaction mix after 10 min, and the sample abs is equal to the absorbance (517 nm) of the sample at the studied concentration.

2.5. Anti-Tyrosinase Activity Evaluation

To evaluate the anti-tyrosinase activity of the different plant extracts obtained by Soxhlet extraction with acetone, the protocol used was based on and adapted from Rangkadilok et al. (2007) [21].
When working with tyrosinase, all reagents and solutions were kept on an ice bath to avoid any loss of activity due to temperature-linked enzyme degradation. A 625 U/mL tyrosinase solution was obtained by adding 4 mL of K3PO4 0.05 M pH 6.5 buffer directly into the commercial vial containing the enzyme (25.000 U) and by vortexing it for 10 s. One mL was retrieved and diluted in the same buffer to reach the required concentration. The total solution was divided into 1 mL aliquots stored at −22 ± 1 °C until used. The aliquots were only thawed once. A 1 mM L-DOPA solution was prepared using distilled water. For this analysis, the inhibition control was a 1 mM kojic acid solution in distilled water, and the blank was pure K3PO4 0.05 M pH 6.5 buffer. As for the lipoxygenase evaluation, pure ethyl alcohol was used for the sample preparation. Five mL of pure ethyl alcohol was added to the dry extracts and submitted to sonication. The solution was then filtered using a 0.45 µm PTFE syringe filter (Whatman, Puradisc 13, Maidstone, UK) and submitted to successive dilutions (from 10 to 10,000 times) in K3PO4 0.05 M pH 6.5 buffer. As oxygen is a co-substrate for the reaction, the buffer was oxygenated for 30 min at room temperature prior to the observation of the reaction.
The enzymatic reaction was performed by mixing 100 µL of sample/blank/control with 50 µL of tyrosinase and 250 µL of the oxygenated buffer. The mixture was left to incubate for 15 min at room temperature and then added into a quartz absorption cell along with 400 µL of the substrate solution. The synthesis of dopachrome was observed in real time, and absorbance was measured every 15 s for 5 min at 475 nm. Each sample/control/blank was carried out in triplicate. The relative activity was obtained according to Equation (4):
%   inhibition   activity = Blank   Slope     Sample   Slope   Blank   Slope   ×   100
where the blank slope is the slope of the blank (0% inhibition) at the linear part of the reaction, and the sample slope is the slope of the samples at the linear part of the reaction, defined by an R2 higher than 0.9.

2.6. Statistical Analysis

For enzymatic inhibition assays, the relative activity was determined using the absorbance slope at its maximum intensity. For the DPPH-reducing potency evaluation assay, the relative activity was based on the absorbance variation after a 30-min reaction period at room temperature. In that case, the relative activity was obtained when comparing the positive control (TROLOX) with the sample results. Each assay was carried out in triplicate. For each activity and dilution, all samples were compared two by two using Tukey’s grouping test (Statistical software Minitab 19 Minitab, State College, PA, USA) with the following parameters: the null hypothesis was that all averages were equal, the significance limit was α = 0.05, the confidence interval was bilateral, and the error rate for the comparison was 5. This allowed the identification of samples or groups of samples showing significantly higher relative activities in comparison to the sample batch.

3. Results and Discussion

3.1. Anti-Lipoxygenase Activity Evaluation

A total of 77 out of the 89 collected plant samples were tested for their ability to inhibit lipoxygenase activity, which therefore indicates their anti-inflammatory potential. The remaining samples were not included in this assay as they showed a strong absorbance at 234 nm. Serial dilutions of plant extracts were performed, allowing us to determine which species presents the highest relative activity and, therefore, which species contains the highest amount of active compounds. Moreover, species showing results above the limit of quantification (LOQ) up to the 104 dilutions are expected to contain several active compounds in their crude extracts or one compound with a high degree of efficiency. The LOQ was established using the relative standard deviation (% RSD) of control replicates immediately after the reagents were thawed. Those %RSD values were not higher than 10%, which is in concordance with standard procedures [22,23,24].
Out of the 77 samples, 17 absorbed too much at the first dilution but entered the evaluation process starting at the second dilution. Results showed that 27 plant extract samples were able to inhibit the lipoxygenase activity above the LOQ at a dilution of 101. Out of those 27 + 17 samples, 9 were still above the LOQ at a dilution of 102. Three of these were still above LOQ at a dilution of 103, and only 1 sample remained above the LOQ at a dilution of 104 (Table 2). Results (Figure 1 and Table 2) highlighted that the 27 plant extracts investigated for their anti-lipoxygenase activity at a dilution of 101 can be classified into 17 different groups. Some of these extracts displayed strong anti-lipoxygenase activities up to 99.05%, and 2 species particularly stand out as being only classified in the group with the highest relative activity (group A): Acalypha hispida dried flower (99.05% ± 4.22) and Erythroxylum corymbosum dried leaf (98.83% ± 2.91). Results (Table 2) also showed that samples with the highest relative activities, which are classified in group A even if there is some crossing over up to group D, ranged from 90.64% to 99.05% of relative activity. These samples can be considered the most effective ones. For the plant extracts diluted 102 times, there is a drop-in activity implying a dose-dependent response as only 9 samples were above the LOQ. Out of these 9 samples, 5 groups were observed. The first group (group A) contained two samples with the following activities: 68.73% ± 3.18 (Zingiber zerumbet dried leaf) and 62.84% ± 1.26 (Zingiber zerumbet dried flower). Interestingly enough, samples from groups A and B only came from one plant species, Zingiber zerumbet, but from different organs of that plant. When looking at the plant extracts diluted 103 times, only 3 samples remained, which were classified into 2 groups, A and B. At this level of dilution, the only sample that was classified in group A was zingiber zerumbet fresh rhizome with 55.73% ± 7.40 of relative activity. Among the samples from this dilution, one that was not considered in the most effective category in the two first dilutions still showed activity over the LOQ: Acalypha hispida dried leaf. Interestingly, only that extract remained above the LOQ with 32.62% ± 2.58 of relative activity at a dilution of 104.
Through the different dilutions, a dose-response change can be observed for most plant extract samples (Figure 1) in their relative activities, except for Acalypha hispida dried leaves. This specific sample is indeed among the most active ones in the first dilution, though it then loses more than 50% of relative activity at the second dilution. It then remains around 30% of activity for the second, third and fourth dilutions as if the inhibition was caused by several compounds, some of these loosing activity as the dilution increases, while others are not impacted by the dilution. There is no simple explanation for these results, as the study of the enzyme inhibition potency of crude plant extracts containing many different molecules is not an easy process. Indeed, these molecules have many interaction options throughout the whole analysis process. They can react with one another but also with the substrate or with the enzyme. In the case of an enzyme interaction, several inhibition processes exist (competitive inhibition, noncompetitive inhibition, uncompetitive inhibition and mixed inhibition). Depending on the type of inhibition, the maximal speed of the reaction, the affinity between the enzyme and its substrate or both can be affected. As the inhibition evaluation was evaluated through kinetic observations, the inhibition phenomenon can also be affected by the incubation time. In addition, it can be due to other regulation modes and not be specifically linked to the enzyme inhibition process. All these phenomena render the study of inhibition arduous, and this is even more so when using an inhibition media as complex as a plant crude extract. This can lead to unexpected observations, such as the one observed for Acalypha hispida where the dilution does not seem to affect the results. Only an in-depth study of the different inhibition mechanisms occurring for this specific sample could help understand such a phenomenon. It is primordial to limit the uses of these results as a primary indication in a screening context.

3.2. Antioxidant Activity (DPPH Test)

The antioxidant properties of the collected plant samples were evaluated through the DPPH test. The average values were compared two by two and grouped using Tukey’s test. %RSD on replicates were not higher than 10%, which is in concordance with standard procedures [22,23,24].
For plant extracts diluted 101 times, 77 out of the 89 samples were above the LOQ. One could not be analyzed at this specific concentration as it impeded with the detector at the given wavelength (517 nm); the latter was re-integrated in the measurements for the following dilutions. These 77 samples were classified into 25 different groups (Table 3), and the group with the highest activity (Group A) showed activities ranging from 94.00% to 99.97%. Only one sample was only classified in that group: Erythroxylum corymbosum dried leaf (99.97% ± 0.06), as the other samples in group A presented some crossing over up to group L. At that dilution, a total of 26 samples were classified in Group A, which can be explained by the presence of compounds with known antioxidant properties, such as polyphenols, in many plants. The second and third dilutions will therefore allow a more precise classification of these samples.
Out of the initial samples and with the addition of the one removed from the plant extracts diluted 10 times, 56 plant extracts diluted 102 times remained above the LOQ. These samples were divided into 23 groups, and 19 samples were classified into group A, showing activities ranging from 92.58% to 98.76%. Only one sample was just in group A: Acalypha hispida fresh leaf (98.76% ± 1.08), as the others presented some crossing over up to group G (activities ranging from 86.63% up to 92.95%).
When looking at the plant extracts diluted 103 times, 21 samples remained above the LOQ and were classified into 9 groups. The group with the highest activities (Group A) contained 4 samples, of which 2 were only in group A: Leea guineensis dried leaf (45.91% ± 3.89) and Litchi chinensis dried leaf (43.10% ± 2.25). The other samples in group A presented some crossing over up to group B (activities ranging from 37.80% to 42.39%). A total of 5 samples were gathered among those high-relative-activity groups: Lawsonia inermis dried leaf, Acalypha wilkesiana fresh leaf, Litchi chinensis dried root, Litchi chinensis dried leaf, and Leea guineensis dried leaf (Figure 2).
Finally, no sample diluted 104 times showed results above the LOQ.
The present results, therefore, allowed the identification of plant species of interest for the isolation of antioxidant molecules. However, it should be noted that other tests could also be considered because of the complexity of oxidative stress and especially what might happen in the human organism.

3.3. Anti-Tyrosinase Activity

The anti-tyrosinase activity of the collected plant samples was evaluated through the observation of tyrosinase inhibition capacity. The activity averages of all samples were compared two by two and grouped using Tukey’s test. The observed limit %RSD to establish the validity of the results was based in concordance with standard procedures, this being a %RSD of 10 [22,23,24]. Out of the 89 samples tested at four different dilutions (samples were respectively diluted 101, 102, 103 and 104 times), 6 samples could not be tested using the anti-tyrosinase protocol, as even when strongly diluted, they strongly absorbed at 475 nm, impeding an accurate reading.
For the 52 tested sample extracts diluted 101 times, Tukey’s test highlighted 22 different groups above the LOQ (Table 4). Five samples were classified into the group with the best activity (Group A) and ranged from 89.74% to 98.57% of relative activity. Only one sample was only in this group: Litchi chinensis dried leaf (98.57% ± 7.09), as the others presented some crossing over up to group D (activities ranging from 80.07% to 89.82%). Together these groups counted for a total of 11 samples with interesting anti-tyrosinase potential.
When the samples were diluted 102 times, 7 samples remained above the LOQ. These samples were divided into 4 groups, and only one sample was classified into the group with the best activity (Group A): Litchi chinensis dried leaf (51.36% ± 3.00). Group B contained Leea guineensis dried fruit (45.45% ± 2.08) and Litchi chinensis dried root (42.29% ± 3.11). Groups C and D had activities ranging from 16.26% to 27.49%. The two other dilutions did not allow for any significant observations. The evolution of the activity of the most active sample is shown in Figure 3.

3.4. Comparison of the Different Activities

In the present work, different organs of different plant species collected in Mayotte were tested for their in vitro anti-lipoxygenase, DPPH-reducing and anti-tyrosinase activities. While the results presented above showed the high potential of certain plant extracts for a single specific activity, the results (Table 5) also highlighted some plant samples displaying more than one interesting activity. Such observations may help in the selection of species for further works. Based on the limit of the analytical methods, the samples showing the best potency for a specific activity were compared with the two other activities.
Therefore, the tested plants can be classified in two categories: the ones that have one strong biological activity, such as Lawsonia inermis and Zingiber zerumbet, and the others that show multiple biological properties. In this second category, different plants can be highlighted, such as Acalypha hispida and Litchi chinensis. The plant species presented here are of high interest for the isolation of novel natural molecules from which originates the determined in vitro activities and that may be incorporated into cosmetic products. Interestingly, Saive et al. (2020) already identified a proanthocyanidin that is responsible for the anti-tyrosinase and antioxidant properties of Litchi chinensis root. In the same way, different compounds, notably belonging to the procyanidin and catechin families, were previously highlighted as interesting antioxidant molecules in Persea americana pit [25]. In addition, zerumbone, a sesquiterpenoid, has already been identified in the fresh rhizome of Zingiber zerumbet and is recognized for its anti-inflammatory properties [26]. By continuing in this perspective, new molecules could be identified and valorized for health purposes.
Table 5. Recapitulative table presenting a comparison of the extracts obtained from different plant species and organs for their investigated biological activities. The comparison is based on the plant organ extracts diluted 100 times for the 3 activities. + the plant organ extract has between 10% and 30% activity, ++ the plant organ extract has an activity superior to 30%, +++ the plant organ extract has an activity that is among the 3 best at the highest dilution, n/a means not applicable.
Table 5. Recapitulative table presenting a comparison of the extracts obtained from different plant species and organs for their investigated biological activities. The comparison is based on the plant organ extracts diluted 100 times for the 3 activities. + the plant organ extract has between 10% and 30% activity, ++ the plant organ extract has an activity superior to 30%, +++ the plant organ extract has an activity that is among the 3 best at the highest dilution, n/a means not applicable.
SampleStatus of the Plant
Species in Mayotte
Anti-Lipoxygenase ActivityDPPH-Reducing AgentAnti-Tyrosinase Activity
Acalypha hispida dried flowerCultivated [27]++++n/a
Acalypha hispida dried leaf+++++n/a
Acalypha wilkesiana dried leafCultivated [27]n/a++n/a
Acalypha wilkesiana fresh leafn/a+++
Cananga odorata dried leafCultivated [10]n/a++n/a
Cananga odorata fresh flowern/a++n/a
Erythroxylum corymbosum dried leafIndigenous [28]n/a++n/a
Lantana camara dried leafExotic [10]n/a++n/a
Lawsonia inermis dried leafCultivated [10]n/a++n/a
Leea guineensis dried fruitIndigenous [10]n/a+++++
Leea guineensis dried leafn/a+++n/a
Litchi chinensis dried leafCultivated [10]+++++++
Litchi chinensis dried rootn/a++++++
Persea americana dried rootCultivated [10]n/a+++
Persea americana fresh pitn/a+++
Persea americana dried pit+++n/a
Zingiber zerumbet dried flowerExotic [10]++++n/a
Zingiber zerumbet dried leaf++n/an/a
Zingiber zerumbet dried stem++n/an/a
Zingiber zerumbet fresh rhizome+++n/an/a

4. Conclusions

On the basis of previous ethnobotanical research in Mayotte, 21 different plants were selected for their potential as skin care agents. Different organs of these were collected and tested in vitro for their capacity to reduce the stable free radical molecule DPPH to inhibit the activity of tyrosinase and/or lipoxygenase in order to better understand why these plants are used in traditional medicine and to discover new biological sources of natural skin care agents.
When looking at the perspectives for future research, two interesting facts stood out. Firstly, 10 promising species were identified based on their in vitro biological activity for skin care. Among these species, some showed yet-unknown biological activities. These specific species could enter future research targeting activities such as topical anti-inflammatory activities. Such plants would be of interest in the development of treatments against dermatitis/eczema and could possibly help to replace the common use of corticosteroids. Secondly, some other species did not show an outstanding specific activity but an average potency for several activities, leading to more generalist applications such as anti-ageing and pigmentation issues or for a relieving balm. When looking at these two points of view, both are of interest: a strong, unique activity or a mild intensity but for multiple activities.
In conclusion, this study highlighted the interesting potential of many plants from Mayotte for their incorporation in cosmetic formulations. Among the 10 plant species of high interest highlighted in the present work, 6 are cultivated in Mayotte, 2 are indigenous to this area, and 2 are exotic. Further studies are now needed, including in vivo tests, in order to confirm the activities shown here in vitro. Additionally, toxicological tests also should be conducted to ensure their innocuity. More studies must also be initiated to identify other molecules of interest. Moreover, if some of these plants begin to be used extensively for their properties, it would also be necessary to set up conservation measures in order to avoid their disappearance. In that case, the target collected organ of the plant will be very important as the collection of fruits, as an example, will not lead to species disappearance, while more attention should be paid when roots or leaves are collected.

Author Contributions

Conceptualization, M.S. and M.-L.F.; Data curation, M.G., L.L., M.S. and C.M.; Formal analysis, M.S. and C.M.; Funding acquisition, M.-L.F.; Investigation, M.S. and C.M.; Methodology, M.S. and M.-L.F.; Project administration, M.-L.F.; Resources, M.-L.F.; Supervision, M.-L.F.; Validation, M.-L.F.; Visualization, M.G., L.L. and M.S.; Writing—original draft, M.G., L.L. and M.S.; Writing—review and editing, M.G., L.L. and M.-L.F. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data is contained within the article.

Acknowledgments

The authors would like to thank Danny Trisman and Thomas Bertrand for their efficient technical support.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Evolution of the relative lipoxygenase inhibition activity of the most efficient plant extract samples through 4 different dilutions (10, 100, 1000 and 10,000). Each column represents the mean ± SD (n = 3) per sample.
Figure 1. Evolution of the relative lipoxygenase inhibition activity of the most efficient plant extract samples through 4 different dilutions (10, 100, 1000 and 10,000). Each column represents the mean ± SD (n = 3) per sample.
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Figure 2. Evolution of the relative DPPH-reducing capacity of the five most efficient samples at 3 different dilutions (101, 102 and 103). Each column represents the mean ± SD (n = 3) per sample.
Figure 2. Evolution of the relative DPPH-reducing capacity of the five most efficient samples at 3 different dilutions (101, 102 and 103). Each column represents the mean ± SD (n = 3) per sample.
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Figure 3. Evolution of the relative tyrosinase inhibition activities of the most efficient samples through 2 different dilutions of 10 and 100 that were above the LOQ. Each column represents the mean ± SD (n = 3) per sample.
Figure 3. Evolution of the relative tyrosinase inhibition activities of the most efficient samples through 2 different dilutions of 10 and 100 that were above the LOQ. Each column represents the mean ± SD (n = 3) per sample.
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Table 1. Recapitulative table presenting the acetone extracts obtained from the different plant species and organs with the biological activities investigated.
Table 1. Recapitulative table presenting the acetone extracts obtained from the different plant species and organs with the biological activities investigated.
SpeciesPlant OrgansIn Vitro Tests
Anti-Lipoxygenase ActivityDPPH-Reducing
Potency
Anti-Tyrosinase Activity
Acalypha hispidaDried flowerxxx
Dried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Acalypha wilkesianaDried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Adansonia digitataDried seedxxx
Dried woodxxx
Aloes mayottensisMucilagexxx
Dried leafxxx
Fresh leafxxx
Cananga odorataDried flowerxxx
Dried leafxxx
Dried rootxxx
Dried woodxxx
Fresh flowerxxx
Erythroxylum corymbosumDried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Erythroxylum lanceumDried leafxxx
Dried root x
Dried woodxxx
Fresh leafxxx
Kalanchoe pinnataDried leafxxx
Dried rootxxx
Dried stemx x
Fresh leaf xx
Lantana camaraDried fruitxxx
Dried leafxx
Dried root xx
Dried woodxxx
Fresh leafxxx
Lawsonia inermisDried leaf xx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Leea guineensisDried fruitxxx
Dried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Litchi chinensisDried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Litsea glutinosaDried leafxxx
Dried rootxxx
Dried wood x
Fresh leafxx
Myristica fragranceDried seedxxx
Pandanus mayottensisDried aerial rootxxx
Dried fruitxxx
Dried leaf xx
Dried woodxxx
Fresh fruitxxx
Fresh leafxxx
Paullinia pinnataDried aerial rootxxx
Dried leaf xx
Dried lianaxxx
Dried rootxxx
Fresh leafxxx
Persea americanaDried leafxxx
Dried pitxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Fresh pitxxx
Sesamum indicumDried seedsxxx
Syzygium aromaticumDried leaf xx
Dried root xx
Dried wood xx
Fresh leaf xx
Tamarindus indicaDried leafxxx
Dried rootxxx
Dried woodxxx
Fresh leafxxx
Zingiber zerumbetDried flowerxxx
Dried leafxxx
Dried rhizom xx
Floral waterxxx
Flower stemxxx
Fresh flowerxxx
Fresh leafxxx
Fresh rhizomxxx
Stemxxx
Table 2. Lipoxygenase inhibition activity results for the plant extracts diluted 10, 100 and 1000 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
Table 2. Lipoxygenase inhibition activity results for the plant extracts diluted 10, 100 and 1000 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
SamplesLipoxygenase Inhibition Activity (%)Grouping Information Based on Tukey’s Method
Dilution 10
Acalypha hispida dried flower99.05A
Erythroxylum corymbosum dried leaf98.83A
Acalypha wilkesiana dried leaf96.70AB
Zingiber zerumbet dried leaf92.36ABC
Cananga odorata fresh flower90.64ABCD
Zingiber zerumbet dried flower88.92 BCD
Zingiber zerumbet stem88.66 BCDE
Lantana camara dried leaf85.19 CDEF
Cananga odorata dried leaf82.82 DEFG
Leea guineensis dried root79.94 EFGH
Acalypha hispida dried leaf78.51 FGHI
Erythroxylum lanceum dried leaf74.49 GHI
Litchi chinensis dried wood72.50 HI
Zingiber zerumbet fresh rhizome70.85 IJ
Cananga odorata dried root62.69 JK
Zingiber zerumbet fresh leaf60.23 KL
Lawsonia inermis dried root58.54 KL
Persea americana dried pit51.57 LM
Zingiber zerumbet flower stem45.04 MN
Litsea glutinosa dried root42.55 N
Paullinia pinnata dried root37.88 NO
Zingiber zerumbet fresh flower33.70 OP
Acalypha hispida dried root33.62 OP
Erythroxylum corymbosum fresh leaf31.22 OP
Acalypha wilkesiana dried root30.30 OP
Paullinia pinnata dried liana27.87 P
Tamarindus indica dried root9.21 Q
Dilution 100
Zingiber zerumbet dried leaf68.73A
Zingiber zerumbet dried flower62.84A
Zingiber zerumbet stem55.90 B
Myristica fragrance dried seed38.19 C
Zingiber zerumbet fresh rhizome32.03 CD
Acalypha hispida dried flower31.15 D
Acalypha hispida dried leaf26.54 D
Litchi chinensis dried leaf16.18 E
Persea americana dried pit13.92 E
Dilution 1000
Zingiber zerumbet fresh rhizome55.73A
Acalypha hispida dried leaf26.13 B
Zingiber zerumbet dried flower21.81 B
Table 3. Antioxidant activity results (DPPH assay) for the plant extracts diluted 10, 100 and 1000 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
Table 3. Antioxidant activity results (DPPH assay) for the plant extracts diluted 10, 100 and 1000 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
SamplesAntioxidant
Activity (%)
Grouping Information Based on Tukey’s Method
Dilution 10
Erythroxylum corymbosum dried leaf99.97A
Acalypha hispida fresh flower99.96AB
Erythroxylum lanceum dried root99.94AB
Lantana camara fresh leaf99.91AB
Erythroxylum corymbosum dried wood99.89AB
Erythroxylum corymbosum fresh leaf99.68ABC
Acalypha hispida fresh leaf99.21ABCD
Erythroxylum lanceum dried leaf99.04ABCDE
Acalypha wilkesiana dried root97.83ABCDEF
Erythroxylum lanceum dried wood97.36ABCDEFG
Myristica fragrans dried seed97.02ABCDEFGH
Acalypha wilkesiana fresh leaf96.49ABCDEFGHI
Cananga odorata dried root96.35ABCDEFGHI
Paullinia pinnata dried leaf96.29ABCDEFGHI
Cananga odorata dried flower96.10ABCDEFGHIJ
Adansonia digitata dried wood95.55ABCDEFGHIJK
Leea guineensis dried root94.86ABCDEFGHIJKL
Syzygium aromaticum fresh leaf94.86ABCDEFGHIJKL
Leea guineensis dried fruit94.51ABCDEFGHIJKL
Persea americana dried root94.50ABCDEFGHIJKL
Paullinia pinnata dried liana94.50ABCDEFGHIJKL
Litchi chinensis dried root94.28ABCDEFGHIJKL
Acalypha hispida dried wood94.28ABCDEFGHIJKL
Leea guineensis fresh leaf94.25ABCDEFGHIJKL
Kalanchoe pinnata fresh leaf94.21ABCDEFGHIJKL
Litchi chinensis dried wood94.00ABCDEFGHIJKL
Persea americana dried pit93.68 BCDEFGHIJKL
Paullinia pinnata dried aerial root93.50 CDEFGHIJKL
Lawsonia inermis fresh leaf93.33 DEFGHIJKL
Lawsonia inermis dried leaf93.27 DEFGHIJKL
Pandanus mayottensis dried fruit93.14 DEFGHIJKL
Syzygium aromaticum dried wood93.05 DEFGHIJKL
Paullinia pinnata dried root92.91 EFGHIJKLM
Litchi chinensis dried leaf92.64 FGHIJKLM
Lantana camara dried leaf92.40 FGHIJKLM
Leea guineensis dried leaf92.31 FGHIJKLM
Litchi chinensis fresh leaf92.19 FGHIJKLM
Leea guineensis dried wood92.18 FGHIJKLM
Persea americana fresh pit91.99 FGHIJKLMN
Paullinia pinnata fresh leaf91.90 FGHIJKLMN
Persea americana fresh leaf91.87 FGHIJKLMN
Litsea glutinosa dried bark91.31 GHIJKLMN
Pandanus mayottensis fresh fruit91.05 HIJKLMN
Acalypha wilkesiana dried leaf90.98 HIJKLMN
Persea americana dried leaf90.95 HIJKLMN
Cananga odorata dried leaf90.87 HIJKLMN
Lawsonia inermis dried root90.31 IJKLMN
Lawsonia inermis dried wood90.30 IJKLMN
Acalypha hispida dried flower89.82 JKLMN
Cananga odorata fresh flower89.78 KLMN
Zingiber zerumbet fresh flower89.61 KLMN
Persea americana dried wood88.91 LMNO
Litsea glutinosa fresh leaf86.66 MNOP
Tamarindus indica dried leaf85.80 NOPQ
Kalanchoe pinnata dried leaf82.38 OPQR
Tamarindus indica fresh leaf80.39 PQR
Acalypha hispida dried leaf80.13 QR
Litsea glutinosa dried leaf78.64 R
Pandanus mayottensis dried root64.81 S
Litsea glutinosa dried wood62.24 S
Litsea glutinosa dried root60.13 S
Zingiber zerumbet dried leaf59.18 ST
Tamarindus indica dried wood58.58 ST
Aloes mayottensis dried root53.21 TU
Lantana camara dried fruit51.92 U
Erythroxylum lanceum fresh leaf50.25 U
Zingiber zerumbet dried flower49.18 U
Pandanus mayottensis dried leaf47.57 U
Aloes Mayottensis dried leaf31.24 V
Zingiber zerumbet fresh rhizome26.45 VW
Pandanus mayottensis fresh leaf21.87 WX
Zingiber zerumbet fresh leaf21.67 WX
Kalanchoe pinnata dried root18.23 XY
Cananga odorata dried wood16.65 XY
Lantana camara dried root16.39 XY
Acalypha wilkesiana dried wood15.35 Y
Lantana camara dried wood12.50 Y
Dilution 100
Acalypha hispida fresh leaf98.76A
Erythroxylum corymbosum fresh leaf98.54AB
Acalypha hispida dried flower96.86ABC
Erythroxylum corymbosum dried leaf95.77ABCD
Acalypha wilkesiana dried leaf94.99ABCDE
Leea guineensis dried fruit94.61ABCDEF
Leea guineensis dried root94.55ABCDEF
Syzygium aromaticum fresh leaf94.36ABCDEF
Leea guineensis fresh leaf93.92ABCDEF
Lawsonia inermis fresh leaf93.91ABCDEF
Acalypha wilkesiana dried root93.71ABCDEF
Litchi chinensis dried wood93.47ABCDEF
Persea americana dried root93.34ABCDEF
Acalypha wilkesiana fresh leaf93.30ABCDEF
Acalypha hispida fresh flower93.27ABCDEF
Litchi chinensis dried root93.26ABCDEF
Litchi chinensis dried leaf92.95ABCDEFG
Leea guineensis dried leaf92.76ABCDEFG
Persea americana dried leaf92.58ABCDEFG
Persea americana fresh pit92.32 BCDEFG
Lawsonia inermis dried leaf92.31 BCDEFG
Syzygium aromaticum dried wood91.79 CDEFG
Lantana camara dried leaf89.71 DEFG
Paullinia pinnata dried root89.39 DEFG
Paullinia pinnata dried leaf89.20 EFG
Persea americana fresh leaf89.12 EFG
Acalypha hispida dried leaf88.36 FGH
Lawsonia inermis dried root86.63 GHI
Paullinia pinnata dried aerial root82.12 HI
Cananga odorata dried leaf81.67 I
Paullinia pinnata dried liana73.33 J
Persea americana dried pit65.31 K
Litchi chinensis fresh leaf62.58 K
Lantana camara fresh leaf55.48 L
Erythroxylum lanceum dried wood52.26 LM
Adansonia digitata dried wood49.57 LM
Tamarindus indica dried leaf48.75 M
Kalanchoe pinnata fresh leaf40.24 N
Erythroxylum corymbosum dried wood38.45 NO
Erythroxylum lanceum dried root37.71 NOP
Persea americana dried wood36.16 NOP
Cananga odorata fresh flower35.01 NOP
Myristica fragrans dried seed33.76 OP
Paullinia pinnata fresh leaf31.38 PQ
Leea guineensis dried wood26.62 QR
Pandanus mayottensis fresh fruit23.15 R
Kalanchoe pinnata dried leaf22.45 R
Pandanus mayottensis dried fruit20.57 RS
Erythroxylum corymbosum dried root20.55 RST
Litsea glutinosa fresh leaf14.33 STU
Pandanus mayottensis dried root13.67 TU
Aloes Mayottensis dried leaf12.01 UV
Erythroxylum lanceum fresh leaf9.69 UV
Pandanus mayottensis dried leaf8.07 UVW
Zingiber zerumbet dried flower6.81 VW
Cananga odorata dried wood3.13 W
Dilution 1000
Leea guineensis dried leaf45.91A
Litchi chinensis dried leaf43.10A
Litchi chinensis dried root42.39AB
Acalypha wilkesiana fresh leaf41.78AB
Lawsonia inermis dried leaf37.80 BC
Syzygium aromaticum fresh leaf36.53 C
Acalypha hispida dried flower35.97 C
Acalypha hispida fresh leaf27.45 D
Acalypha hispida fresh flower26.83 D
Leea guineensis fresh leaf26.48 DE
Lawsonia inermis fresh leaf26.38 DE
Leea guineensis dried fruit26.07 DE
Persea americana dried leaf26.02 DE
Erythroxylum corymbosum dried leaf22.67 DEF
Tamarindus indica dried leaf21.83 EF
Syzygium aromaticum dried wood18.49 FG
Acalypha wilkesiana dried leaf17.86 FG
Acalypha hispida dried leaf16.39 G
Persea americana dried pit7.81 H
Erythroxylum corymbosum dried wood3.06 HI
Leea guineensis dried wood2.65 I
Table 4. Anti-tyrosinase activity results for the plant extracts diluted 10 and 100 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
Table 4. Anti-tyrosinase activity results for the plant extracts diluted 10 and 100 times (n = 3). Results were statistically analyzed using Tukey’s grouping test (confidence level: 95%), and the observed group classifications (A, B, C……etc.) are shown in the present table.
SamplesAnti-Tyrosinase
Activity (%)
Grouping Information Based on Tukey’s Method
Dilution 10
Litchi chinensis dried leaf98.57A
Leea guineensis dried fruit93.28AB
Leea guineensis fresh leaf91.53ABC
Leea guineensis dried root89.82ABCD
Syzygium aromaticum dried leaf89.74ABCD
Lawsonia inermis dried root87.69 BCD
Syzygium aromaticum fresh leaf86.22 BCD
Litchi chinensis dried root84.83 BCDE
Persea americana dried leaf82.05 CDE
Persea americana dried root80.10 DE
Lawsonia inermis dried leaf80.07 DE
Litchi chinensis dried wood75.61 EF
Paullinia pinnata dried leaf68.99 FG
Kalanchoe pinnata fresh leaf66.18 FGH
Adansonia digitata dried wood64.91 GHI
Paullinia pinnata dried root63.91 GHI
Persea americana fresh leaf62.84 GHI
Litchi chinensis fresh leaf62.36 GHI
Persea americana fresh pit60.80 GHIJ
Acalypha hispida fresh leaf60.20 GHIJ
Lawsonia inermis fresh leaf59.63 GHIJ
Acalypha wilkesiana fresh leaf59.01 HIJ
Paullinia pinnata dried aerial root56.23 HIJK
Lantana camara fresh leaf54.99 IJK
Paullinia pinnata dried liana51.78 JKL
Persea americana dried pit51.64 JKL
Tamarindus indica fresh leaf51.33 JKL
Erythroxylum lanceum dried wood48.76 KLM
Persea americana dried wood47.22 KLMN
Litsea glutinosa dried root44.81 LMN
Pandanus mayottensis dried aerial root44.07 LMNO
Erythroxylum corymbosum dried root43.89 LMNOP
Acalypha hispida dried flower43.69 LMNOPQ
Syzygium aromaticum dried wood43.47 LMNOPQ
Cananga odorata fresh flower40.55 MNOPQR
Acalypha hispida dried root40.17 MNOPQR
Aloes mayottensis mucilage39.15 MNOPQRS
Zingiber zerumbet dried rhizome38.77 NOPQRS
Paullinia pinnata fresh leaf34.12 OPQRST
Tamarindus indica dried root34.08 PQRST
Cananga odorata dried root33.85 QRST
Zingiber zerumbet fresh leaf31.72 RSTU
Zingiber zerumbet flower stem30.59 RSTU
Zingiber zerumbet stem29.61 STU
Erythroxylum corymbosum dried wood29.40 STU
Syzygium aromaticum dried root28.46 TUV
Cananga odorata dried wood27.51 TUV
Zingiber zerumbet fresh rhizome26.71 TUV
Zingiber zerumbet floral water26.14 TUV
Zingiber zerumbet dried flower24.76 TUV
Pandanus mayottensis fresh leaf22.70 UV
Aloes mayottensis dried leaf18.85 V
Dilution 100
Litchi chinensis dried leaf51.36A
Leea guineensis dried fruit45.45 B
Litchi chinensis dried root42.29 B
Persea americana dried root27.49 C
Syzygium aromaticum dried leaf27.28 C
Persea americana fresh pit18.69 D
Acalypha wilkesiana fresh leaf16.26 D
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Genva, M.; Lheureux, L.; Saive, M.; Maes, C.; Fauconnier, M.-L. Study of the Cosmetic Potential Uses of Plants from Mayotte as Skin Care Agents through the Screening of Their Biological Activities. Nutraceuticals 2022, 2, 420-440. https://doi.org/10.3390/nutraceuticals2040031

AMA Style

Genva M, Lheureux L, Saive M, Maes C, Fauconnier M-L. Study of the Cosmetic Potential Uses of Plants from Mayotte as Skin Care Agents through the Screening of Their Biological Activities. Nutraceuticals. 2022; 2(4):420-440. https://doi.org/10.3390/nutraceuticals2040031

Chicago/Turabian Style

Genva, Manon, Laura Lheureux, Matthew Saive, Chloé Maes, and Marie-Laure Fauconnier. 2022. "Study of the Cosmetic Potential Uses of Plants from Mayotte as Skin Care Agents through the Screening of Their Biological Activities" Nutraceuticals 2, no. 4: 420-440. https://doi.org/10.3390/nutraceuticals2040031

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