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Pokajewicz, K.; Czarniecka-Wiera, M.; Krajewska, A.; Maciejczyk, E.; Wieczorek, P.P. Biological Activities of Lavandin. Encyclopedia. Available online: https://encyclopedia.pub/entry/43465 (accessed on 20 April 2024).
Pokajewicz K, Czarniecka-Wiera M, Krajewska A, Maciejczyk E, Wieczorek PP. Biological Activities of Lavandin. Encyclopedia. Available at: https://encyclopedia.pub/entry/43465. Accessed April 20, 2024.
Pokajewicz, Katarzyna, Marta Czarniecka-Wiera, Agnieszka Krajewska, Ewa Maciejczyk, Piotr P. Wieczorek. "Biological Activities of Lavandin" Encyclopedia, https://encyclopedia.pub/entry/43465 (accessed April 20, 2024).
Pokajewicz, K., Czarniecka-Wiera, M., Krajewska, A., Maciejczyk, E., & Wieczorek, P.P. (2023, April 25). Biological Activities of Lavandin. In Encyclopedia. https://encyclopedia.pub/entry/43465
Pokajewicz, Katarzyna, et al. "Biological Activities of Lavandin." Encyclopedia. Web. 25 April, 2023.
Biological Activities of Lavandin
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Lavandin essential oil has been found to have antioxidant and biocidal activity (antimicrobial, nematicidal, antiprotozoal, insecticidal, and allelopathic), as well as other potential therapeutic effects such as anxiolytic, neuroprotective, improving sleep quality, antithrombotic, anti-inflammatory, and analgesic.

Lavandula x intermedia Lavandula hybrida Lavandula angustifolia

1. Introduction

Lavandula x intermedia Emeric ex Loisel (LI), also known as lavandin, Dutch lavender, or bastard lavender, is a widely cultivated aromatic plant belonging to the family Lamiaceae Lindl. It is a hybrid of true lavender—Lavandula angustifolia (LA)—and spike lavender—Lavandula latifolia (LL). While it shares many similarities with its parent species, lavandin possesses unique characteristics that set it apart. This entry is a continuation of an article entitled “Lavandula x intermedia—A Bastard Lavender or A Plant of Many Values? Part I. Biology and Chemical Composition of Lavandin” [1]. Part I covered the biological and chemical characteristics of L. x intermedia, including taxonomy, geographical range, morphological features, popular cultivars, cultivation, and essential oil production. Additionally, the chemical composition of its essential oil and hydrolate was thoroughly discussed and compared to the parent species, taking into account the current industry standards such as ISO, European Pharmacopeia (Ph. Eur.), and WHO monographs. Researchers stated that lavandin essential oil (further referred to as lavandin oil) has a similar chemical composition to LA, but with a higher concentration of terpenes that give it a camphor scent, making it less appealing for use in the perfume industry. However, LI has some benefits, such as a higher yield of essential oil and lower production cost, making it a favored lavender crop for farming. Nonetheless, despite its commercial success and widespread cultivation, there is a shortage of scientific research on the subject. The scientific community tends to focus on LA, a raw material recognized by European Pharmacopeia.

2. Biological Activities of Lavandin

The most obvious and apparent biological property of lavandin is its smell. It is caused by the volatile chemicals, mainly oxygenated monoterpenes, secreted and stored in the aerial parts of the plant [2][3][4][5]. Most of the applications of LI in industries and daily life result from this significant feature of this plant. Apart from its smell, lavandin, like many other aromatic herbs, is associated with numerous biological effects.

2.1. Biocidal Activities

2.1.1. Antimicrobial

Many essential oils (EOs) exhibit antimicrobial properties. They have been used for centuries in traditional medicine and for embalming a corpse. Even though multiple EOs have demonstrated antimicrobial action, only some possess the potential to be used as antimicrobial agents. The real-world effect is usually significantly weaker compared to antibiotics and other synthetic compounds [6]. L. angustifolia has been proven to be effective against many bacteria, fungi, and some viruses [6][7][8][9]. There is also multiple evidence for the antibacterial and antifungal action of lavandin oil, but according to the best knowledge—there is not any research investigating its antiviral effect.
Antimicrobial studies of lavandin oil, like other essential oils, are usually conducted in vitro with the use of agar diffusion (disc or well) methods and/or dilution methods. The diffusion methods, especially the disc diffusion method, are mainly used for antimicrobial susceptibility testing. Dilution methods are the most suitable for the determination of minimum inhibitory concentration (MIC), minimal lethal concentration (MLC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) values due to the fact they enable the calculation of the concentration of the tested antimicrobial chemical in the broth or agar media [7][10][11].
The antibacterial and antifungal effect of lavandin EOs against many gram-positive and negative bacteria was demonstrated by multiple researchers. Different lavandin cultivars were tested. For example, Garzoli and coworkers tested EO of the very popular cultivar Grosso grown in Italy against Escherichia coli, Acinetobacter bohemicus, Pseudomonas fluorescens, Bacillus cereus, and Kocuria marina and found bactericidal effect on Gram-negative bacteria and a bacteriostatic effect on Gram-positive bacteria both for the liquid and vapor phases [12]. According to the various tests that authors conducted, A. bohemicus was the most vulnerable strain to lavandin essential oil. It exhibited an inhibition zone of 47 mm (greater than of positive control gentamicin) and had MIC of 0.47% in the broth microdilution test. P. fluorescens was the most resistant among all the strains tested. It had an inhibition zone of just 8.5 mm and MIC of 3.75%. Bajalan et al. found the high antibacterial activity of Iranian lavandin oil from leaves against G− E. coli and G+ Streptococcus agalactiae and moderate against G− K. pneumoniae and S. aureus [13]. The antibacterial effect in vitro and in vivo in mice against Citrobacter rodentium (G−) was also indicated by Baker et al. [14]. When L. x intermedia and L. angustifolia are considered in one study, usually lavandin oil possesses similar or stronger antibacterial and antifungal effects than true lavender oil. Jianu et al. investigated Romanian LI and LA essential oil against Enterococcus faecium, Shigella flexneri, Salmonella typhimurium, Escherichia coli, and Streptococcus pyogenes. The studied oils presented significant bactericidal effects against S. flexneri, S. aureus, and E. coli but not against S. pyogenes. In most cases, L. x intermedia antibacterial activity was higher [15]. Stronger action of LI was also observed by Tardugno and coworkers, who tested EOs of different cultivars of LI and LA (Italian origin) against Listeria monocytogenes [16]. Di Vito et al. indicated similar antibacterial and antifungal properties of both lavender oils with a slightly higher effect for L. intermedia [17]. On the other hand, Robu et al. tested Romanian LI and LA essential oils against S. aureus, S. pyogenes, P. aeruginosa, E. coli, and Candida albicans, and they noticed that L. angustifolia essential oil was more active on certain bacterial strains, but L. x intermedia EO was more effective against Candida [18]. Antifungal properties of LI EO in high doses against Candida albicans were noticed by Karakaş and Bekler [19]. Moon et al. have also observed the antifungal activity of oils of lavandin and other species of lavender they studied. The EOs of three different cultivars of LI and LA oils were effective against Aspergillus nidulans and Trichophyton mentagrophytes [20]. Lavandin oil was also proved by Larrán et al. to be fungistatic against some strains of studied Ascosphaera apis—the fungus causing the chalkbrood disease of bees [21]. However, Erland and coworkers tested LI ‘Provence’ and ‘Grosso’ and LA oils and observed no significant antifungal effect against three agricultural pathogens, except some activity of LI ‘Provence’ oil against B. cinerea [22].
When comparing the antimicrobial activity of lavandin or true lavender oil with the oils of other aromatic plants, it has been found that some plants are far more effective, usually due to their high content of phenolic compounds, which are characterized by strong antimicrobial properties. Tardugno et al. conducted in vitro screening to assess the antimicrobial activity of 14 essential oils against oral pathogenic bacteria. It was indicated that lavandin oils showed moderate activity among all tested oils, with MIC ranging from 2–512 μL/mL. The most effective oils were those derived from Thymus vulgaris and Rosmarinus officinalis, which had MICs of 4–16 and 1–32, respectively [23]. Rota and coworkers studied the antimicrobial activity against selected foodborne pathogenic bacteria. Once again, lavandin oil showed intermediate antibacterial activity among the tested samples. As expected, the biggest effects were observed for T. vulgaris and Satureja montana oils, whereas the weakest effects were noticed for Salvia sclarea and Hyssopus officinalis [24]. The above-mentioned Di Vito et al. also studied other than lavender EOs and found that both lavandin and true lavender oils exhibited weaker activity against tested microorganisms (bacteria, drug-resistant yeasts, and fungal dermatophytes) than oils containing a lot of thymol and/or carvacrol, such as those from Origanum hirthum, S. montana, Monarda didyma, and Monarda fistulosa. The same authors also demonstrated that essential oils work much stronger than hydrolates, which exhibited mostly high MIC and MLC values (above 50%), whereas lavender essential oils had MIC and MLC values mostly above 2% [17]. No antibacterial activity of Lavandula spp. hydrolates was observed by Moon et al. The authors also evaluated aqueous and ethanolic extracts and found that water extract had no activity, while some ethanolic extracts were effective against Proteus vulgaris [20]. Ramić and colleagues tested lavandin essential oil and ethanolic extracts and observed strong antibacterial activity against one of the major food-borne pathogens—Campylobacter jejuni, with EOs exhibiting the strongest effect and MIC of 0.25 mg/mL, whereas ethanolic extracts had MIC of 0.5–1 mg/mL) [25]. The antimicrobial activity of lavandin ethanolic extracts of the same ‘Budrovka’ cultivar was also confirmed by other researchers—Blazenkovic et al., who found that ethanolic extracts, especially those from flowers, exhibited antimicrobial activity against a broad spectrum of bacteria, yeasts, molds, and dermatophytes. The antimicrobial activity of the extracts decreased in the order of plant part: flowers > leaves > inflorescence stalks [26].
Summarizing the above findings, there is no doubt that lavandin preparations, such as essential oil or ethanolic extract, possess antimicrobial activity. This activity is at least as strong as the activity of L. angustifolia or stronger. Hydrolates and other lavandin preparations do not exhibit antimicrobial power, or it is significantly weaker. Regarding the antimicrobial activity of essential oils, even though it is proven and well-established, the therapeutic effect is significantly weaker when compared to synthetic antibiotics. Essential oils are volatile, and their ability for quick vaporization can shorten their effectiveness. On the other hand, this drawback can be at least partially overcome by an appropriate drug formulation [6]. Varona et al. demonstrated that the activity of lavandin oil could be enhanced by encapsulation due to the protection and controlled release of the oil components [27]. According to the literature, antimicrobial activity is usually correlated with phenolic, aromatic, or alcoholic components of essential oils. Their main mechanism of action is related to some disruption of the cell membrane and its increased permeability [28][29][30][31]. The main oil component of L. x intermedia, linalool, can destroy bacterial cell walls, change membrane potential, and enhance membrane leakage [32]. The antibacterial effect of EOs is generally more pronounced in the case of G+ bacteria. It is believed that the rigid outer membrane of G– bacteria limits the diffusion of hydrophobic compounds, therefore, protecting them against the harmfulness of EOs components [10]. However, the presented review of the studies of lavandin oil does not always confirm this general belief. Diverse antibacterial powers were observed regardless of gram-positive or negative attribution of studied bacteria.

2.1.2. Other Biocidal

Articles reporting on the biocidal power of lavandin preparations, other than the antimicrobial power, are not numerous. An antiparasitic activity of lavandin was indicated by Moon and coworkers. The scientists studied the effect of lavandin and true lavender essential oils on three protozoal pathogens: Giardia duodenalis, Trichomonas vaginalis, and Hexamita inflata. They demonstrated that oil concentrations of 1% or less could eliminate pathogens in the culture. L. angustifolia essential oil presented a slightly stronger effect than L. x intermedia. The authors stated: “Whether lavender essential oils can be used as a viable treatment for infected water sources or as a treatment of mammalian and/or fish parasitic infection is unknown. The previously unreported finding that these oils are potent anti-protozoan agents should, however, be further investigated” [33]. According to the best knowledge, there are no further reports on the antiprotozoal activity of L. x intermedia EO.
Lavandin oil was also investigated regarding nematicidal power. D’Addabbo et al. observed a powerful biocidal effect on pathogenic root-knot nematode—Meloidogyne incognita and walnut root lesion nematode—Pratylenchus vulnus. The activity was so high (LC50 equal to 1.2 and 3.1 μg/mL for one essential oil of one LI cultivar) that authors postulated the oil as a component of the new nematicidal formulations alternative to synthetic nematicides. The effectiveness of lavandin oils of three different cultivars was evaluated both in vitro and in vivo in soil. LI oils significantly reduced parasite eggs’ density, their hatchability, and the gall formation on roots, overall positively impacting the growth of the studied tomato plants [34]. A potent nematicidal effect was also documented in vitro and in vivo by Andrés et al. [35]. They studied the lavandin and some other plants’ hydrolates, by-products produced during steam distillation, against the root-knot nematode Meloidogyne javanica. All tested hydrolates showed nematicidal effects in vivo (both on larvae mortality and suppression of egg hatching). The nematicidal active components of lavandin hydrolates were found to be present in the aqueous fraction, indicating that some polar constituents of lavandin, rather than those unpolar, are responsible for the observed effect. This is supported by the lack of nematicidal activity of L. x intermedia essential oil against the same nematode—M. javanica observed by de Elguea-Culebras et al. [36]. No nematicidal effect of lavandin oil was also observed by Park and coworkers against pinewood nematode (Bursaphelenchus xylophilus), but they did not present the detailed results for lavandin it and focused only on three chosen essential oils from other plants [37]. Considering all the above, the effectiveness of the nematicidal efficacy of lavandin is uncertain, and more studies are needed in this field.
The above-described nematodes are loss-making pests in agriculture. The other, an even bigger group of pests, is insects. Therefore, lavandin oils were also tested regarding their insecticidal or repelling properties for potential use as natural-based plant protection agents. The repelling properties of LI essential oils were indicated for the following insects: maize weevil Sitophilus zeamais, rusty grain beetle Cryptolestes ferrugineus, and yellow mealworm beetle Tenebrio molitor [38]. LI hydrosols also exhibited repellency in studies on the confused flour beetle Tribolium confusum [39]. This insect was also studied by Theou and coworkers [40]. They tested lavandin and other essential oils and found out that all tested oils, except oregano oil, exhibited strong toxicity to all developmental stages of the pest. They postulated the EOs as fumigants used for the protection of stored products in storehouses. The insecticidal power of lavandin oils was also recognized by other researchers who showed its efficacy against bean weevil Acanthoscelides obtectus, colorado potato beetle Leptinotarsa decemlineata, and spotted wing drosophila Drosophila suzukii. A positive correlation between total oxygenated monoterpenoid content and insecticidal activity was observed, with linalool and 1,8-cineole being the most effective terpenes [36][41][42].
Essential oils are also generally known for their allelopathic activity. In the case of lavandin oil, according to the knowledge, there are two studies concerning it. Both studies examined the effect of lavandin oil on lettuce Lactuca sativa and English ryegrass Lolium perenne. de Elguea-Culebras et al. showed low to moderate toxicity for the assayed oils in the allelopathic test. The LI essential oil did not show negative effects on the germination of L. sativa but reduced the growth of its root [36]. Santana and coworkers observed some phytotoxic activity against L. sativa and L. perenne seeds and observed negative effects on germination and growth [43]. Regarding hydrolates of lavandin, both hydrolates from flowers and stems were able to inhibit the germination of radish Raphanus sativus, with a stronger effect of the flower hydrolate [39]. Extensive allelopathic studies were conducted by Haig and coworkers, who studied the effects of aqueous extracts of several lavender species, including L. x intermedia [44]. The researchers indicated that L. x intermedia was the most phytotoxic among the tested species. It showed the effect on all four tested plant species. After fractionization of the LI extract, they found that the fraction consisting of coumarin and 7-methoxycoumarin was the most phytotoxic, and the coumarin was largely responsible for the effect. Coumarin is a well-known phytotoxin, and lavandin is known to contain more coumarins than, for example, L. angustifolia [44][45]. This explains the stronger effect of lavandin extracts as a phytotoxic agent.

2.2. Antioxidant Power

Oxidative stress is caused by an imbalance between the production and deactivation of oxygen-reactive species (ROS). ROS are naturally generated as by-products of oxygen metabolism and can play some physiological roles (among others—cell signaling). In stressful environmental conditions and the presence of xenobiotics, the production of ROS increases, leading to the imbalance that causes cell death and some pathologies, such as some cancers or neurodegenerative disorders. Therefore, apart from the endogenous antioxidant systems, also exogenous antioxidants are intensively studied. Antioxidants are compounds capable of slowing or retarding the oxidation of an oxidizable material and protecting organisms from oxidative stress. As synthetic antioxidants such as butylated hydroxyanisole (BHA) or butylhydroxytoluene (BHT) are suspected to be potentially harmful to human health, many natural products, including essential oil and plant extracts, have been investigated for their antioxidant properties [46][47][48].
Antioxidant power is the second, just after biocidal, activity reported in the literature for L. x intermedia. Researchers have found ten original research articles reporting it: three of them relate to essential oil, six to plant solvent extracts, and one to both of the formulations. Most of them relate to in vitro studies based on simple radical scavenging assays. However, there is a report by Hancianu et al., who conducted a detailed study on Wistar rats subjected to scopolamine—an induced rat model of dementia [49]. The week-long inhalation of the essential oils of L. angustifolia and L. x intermedia by rats for a week induced some significant biochemical changes in their brains. Temporal lobe homogenates indicated increased activity of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX), increased content of reduced glutathione (GSH), and reduced malondialdehyde (MDA) level. Rats in both lavender groups exhibited a significant decrease in MDA levels. MDA is one of the final products of polyunsaturated fatty acids peroxidation in the cells. Decreased MDA levels reflected the reduced lipid peroxidation caused by free radicals. Increased activity of CAT, SOD, and GPX, which create an enzymatic endogenous antioxidant defensive system [46], results in less oxidative stress. The results of the experiment show that lavender oils can be an indirect antioxidant. Indirect antioxidants enhance natural antioxidant defenses in living organisms by inducing the expression or increasing the activity of antioxidant enzymes [47]. Additionally, the authors reported that DNA cleavage patterns (present in the scopolamine-alone-treated rat group) were absent in the scopolamine-treated rats exposed to lavender oils, suggesting that lavender oils possess antiapoptotic and neuroprotective activity [49].
The antioxidant activity of lavandin EOs in vitro was indicated by Carrasco and coworkers [50][51]. In one paper, they described the studies of lavandin ‘Abrial’, ‘Super’, and ‘Grosso’ essential oils of Spanish origin. They conducted five different antioxidant assays: ABTS, DPPH, oxygen radical absorbance capacity (ORAC), chelating power (ChP), and reducing power (RdP). Studied L. x intermedia oils showed moderate antioxidant activities with varying activity for different cultivars and different tests. The authors also indicated mild inhibition of lipoxygenase (LOX) in vitro. LOX is a crucial enzyme in the transformation of arachidonic acid into leukotrienes which are involved in the occurrence of inflammation. LOX inhibition can lower leukotriene levels, thereby delivering an anti-inflammatory effect [50][52]. In the other article, Carrasco et al. studied the antioxidant activity and hyaluronidase inhibition of different species of Lavandula and Thymus essential oils. Hyaluronidase inhibitors can potentially serve as anti-inflammatory, anti-microbial agents, and anti-aging agents [53]. However, the researchers did not observe any inhibitory effect of lavandin oil and a very weak effect of LA oil on the enzyme. Regarding antioxidant activity, authors indicated some antioxidant activity of LI oils, in most assays, weaker than for true lavender oil, while both lavender oils showed significantly lower activity than Thymus zygis, which was caused by the high presence of thymol, the compound with known and strong antioxidant properties [51]. In general, phenolic agents act as antioxidants due to their high reactivity with peroxyl radicals, which are disabled by hydrogen atom transfer [47]. Lavender essential oils do not contain any significant amounts of phenolic terpenes and phenylpropanoids. In general, essential oils containing no or little phenols and cyclohexadiene-like components (e.g., γ-terpinene, α-terpinene, and α-phellandrene) do not exhibit significant direct antioxidant potency [47]. Direct antioxidants are compounds able to impair the radical chain reaction causing oxidation. However, despite the low presence of high-impact direct antioxidants, lavandin oil might show an indirect antioxidant property, as indicated by the above-described experiment of Hancianu et al. [49].
One more issue needs to be raised whenever the antioxidant activities of essential oils are assessed based on indirect methods such as the DPPH, ABTS, FRAP, or Folin–Ciocalteu tests. These methods are flawed models of antioxidant properties and are often inappropriate for reliable measuring antioxidant properties of essential oils. For example, a very popular and basic DPPH assay gives positive outcomes for some essential oils, not due to its antioxidant activity, but rather due to binding a hydrogen atom from C–H bonds from terpenes with a sufficiently low bond dissociation enthalpy such as α- or β-pinene and limonene. Therefore, the discoloration of the reactant might not necessarily indicate the antioxidant potency but also the presence of highly oxidizable compounds in the essential oil [47]. Moreover, the results are often difficult to compare due to multiple assays and different and inconsistent units. Thus, the discussion and most of the comparations are usually performed between samples from one experimental setup.
Lavandin ethanolic extracts exhibit higher antioxidant activity than lavandin oil [25]. The superior to EO antioxidant power of the lavandin extract is caused by a different chemical composition compared, namely more abundant flavonoids, coumarins, phenolic acids, and their glycosides [25][45][48][54]. Higher activity was shown for extracts of post-distillation waste than the raw plant material before distillation [25][55]. Regarding the plant part, the most potent were found to be ethanolic extracts of leaves, then flowers and inflorescence stalk [56]. In one experiment, Berrington and Lall studied the antioxidant power of acetone LI and LA extracts, as well as other plant species. They have found that both lavender species had the lowest antioxidant activity (DPPH assay) among tested plant extracts (Origanum vulgare, Rosmarinus officinalis, Thymus vulgaris, Petroselinum crispum, Foeniculum vulgare, and Capsicum annuum) [57]. Whenever both activities of LI and LA were assessed in one study, they were rather similar, with varying antioxidant power depending on the assay and tested cultivar. True lavender extracts contained more phenolic acids and flavonoids, while coumarins were at higher levels in lavandin extracts [45][56][57]. Looking at the gathered studies, researchers cannot claim any superiority of L. angustifolia over L. x intermedia in this field.

2.3. Other Activities

Lavender essential oils are very popular in complementary/alternative medicine and are commonly used in aromatherapy to reduce stress, increase relaxation, and improve the quality of sleep [7][48][58][59]. The anxiolytic action of L. angustifolia oil was proven both in rodents and humans by both inhalation and ingestion. The most substantial evidence comes from the studies of Silexan, a patented active substance comprised of Lavandula angustifolia essential oil produced from flowers, standardized, compliant with European Pharmacopeia, and manufactured by Dr. Willmar Schwabe GmbH & Co. KG in Germany. Oral administration of Silexan impacted positively depressed mood, sleep disturbances, and the overall quality of life of clinical trial participants. It was also demonstrated that sleep improvement is a result of the anxiolytic effect, not the sedative effect per se [60][61][62]. The mechanisms underlying the anxiolytic effects of Silexan are not certain. Action through the mediation of gamma-aminobutyric acid (GABA) was proposed by Aoshima and Hamamoto [63]. Schuwald et al. demonstrated that Silexan inhibited voltage-dependent calcium channels (VOCCs) in neuronal cells at nanomolar concentrations [64]. It has been speculated that under anxiety or stress disorders, an enhanced calcium ions influx through VOCCs could increase the release of neurotransmitters such as glutamate and norepinephrine, which are involved in the pathogenesis of these diseases. Baldinger and coworkers showed that Silexan reduced the 5-HT1A receptor binding potential in the brain clusters such as the temporal gyrus, the fusiform gyrus, the hippocampus, the insula, and the anterior cingulate cortex. This led to an increase in extracellular serotonin content. Most probably, the effect of Silexan, apart from VOCCs inhibition, is additionally mediated via the serotonergic neurotransmitter system, particularly the 5-HT1A receptor, not through a GABAnergic mechanism [60][61][62]. The results of Siloxane might suggest a similar action of lavandin oil. However, there are almost few studies to support it. Hancianu et al. studied the effect of Romanian essential oils from L. x intermedia and L. angustifolia, as well as Siloxan, on a dementia rat model. They found that not only Siloxan but also both studied lavender oils acted neuroprotective and improved spacial memory and performance in various tests suggesting anxiolytic and antidepressant activity [49][65]. Regarding the influence on sleep, one human study with a mixture of lavandin oil with bergamot and ylang-ylang oils was described. The five-day-long aromatherapy improved the subjective assessment of sleep quality in the examined patients in cardiac rehabilitation [66].
Another often-raised biological action of lavender oils is their anti-inflammatory effect. Linalool, the main component of lavender oil, has been reported to have anti-inflammatory effects [7][48][67][68]. Huo and coworkers tested the action of this terpene on lipopolysaccharide(LPS)-induced production of inflammatory mediators such as tumor necrosis factor α (TNF-α) a and interleukin 6 (IL-6) [69]. LPS is a component of the outer membrane of G– bacteria, which triggers a strong immune response. Scientists have found that linalool reduced the production of TNF-α and IL-6 both in stimulated macrophages in vitro and in vivo in lung injury mouse models. They showed that linalool treatment attenuated lung histopathology in mice. In search of molecular mechanisms of the linalool action, the researchers investigated the phosphorylation of some proteins in NF-κB and MAPK pathways. Nuclear factor-kB (NF-κB) is the critical dimer protein controlling the expression of over 500 genes, including many inflammation-associated factors. Many agents and stimuli can activate NF-κB through canonical and noncanonical pathways. Usually, NF-κB is kept in the cytoplasm in inactive form due to its binding with its inhibitor (lκB). After some stimuli, such as LPS, IκB is phosphorylated and later degraded. The released NF-κB is translocated to the nucleus, followed by the p65 subunit phosphorylation, acetylation, methylation, as well as subsequent DNA binding and gene transcription. In this way, nuclear factor-kB activation mediates the activation of proinflammatory genes, including TNF-a and IL-6 [69][70]. Huo and coworkers indicated that linalool blocked LPS-induced IκBα phosphorylation and consequently prevented NF-kB activation. They also noticed reduced phosphorylation of ERK, JNK, and p38 in the MAPK signaling pathway, another effect that led to the anti-inflammatory activity of linalool. The effect of linalool on acute lung inflammation induced by other stress stimuli—cigarette smoke (CS)—was investigated by Ma et al. in vivo in mice [71]. It was indicated that linalool significantly reduced the production of TNF-α, and IL-6, along with some other inflammatory mediators. Overall, it inhibited the infiltration of inflammatory cells and lung inflammation. The researchers noted that the terpene suppressed CS-induced NF-κB activation by inhibiting CS-induced IκBα and p65 NF-κB protein phosphorylation. Therefore, the demonstrated effect of linalool is in agreement with the former research conducted by Huo et al. [69]. This mechanism might be responsible for the reported anti-inflammatory properties of different Lavandula essential oils. Specifically to L. x intermedia, researchers have found a study on this activity—the above-mentioned study of Baker and coworkers on the lavandin oil effect on mice acute colitis induced by Citrobacter rodentium bacteria [14]. The oil administration lowered the expression of inducible nitric oxide synthase, interferon-gamma, interleukin 22, and macrophage inflammatory protein-2α gene expression and decreased neutrophil and macrophage infiltration. In colitic mice, oral gavage with lavandin oil resulted in milder disease, decreased morbidity and mortality, and reduced intestinal tissue damage. Barocelli et al. also noticed the gastroprotective effect of lavandin oil, as well as linalool and linalyl acetate, delivered separately. LI oil administration protected against acute ethanol-induced gastric ulcers in rats, but the mechanism of its protective action was not elucidated [72]. They have also investigated the analgesic effect of chemical and thermic stimuli. Lavandin oil, especially when inhaled, prolonged the response to unpleasant stimuli, suggesting an antinociceptive effect. However, some authors postulated that, instead of having a direct analgesic effect, inhalation of lavender oil may cause a more positive attitude and therefore alter the subjective perception of pain unpleasantness [7][73]. Linalool, the main lavender terpene, was demonstrated to induce analgesic effects in mice—significantly increasing the pain threshold and attenuating pain behaviors. Antinociceptive effects were absent in olfactory-deprived mice in which the olfactory epithelium was damaged. Thus, the action of linalool might be triggered by the olfactory sensory input. An immunohistochemical study revealed that linalool activated hypothalamic orexin neurons, crucial mediators for pain processing. Still, the actual mechanism is not understood [74][75].
The oil administration significantly reduced thrombotic events in mice models of pulmonary thromboembolism variance with aspirin used as a reference drug but without inducing hemorrhagic complications as acetylsalicylic acid. Regarding the potential anticancer properties of lavandin, there are only a few investigations. Berrington et al. studied the acetone extracts of LI and LL on the cervical epithelial carcinoma cell line and observed no anticancer activity [57]. Tabatabaei et al. observed the potential anticancer/antiproliferative activity of lavandin oil on the human breast cancer cell line MCF-7 [76], and Ovidi and colleagues found similar activity on different cancerous cell lines [77]. The latter authors also found that nanoformulation of the essential oil increased its antiproliferative activity.
As already mentioned, the biological effects of lavandin oil, which have been tested and verified, are mostly related to biocidal or antioxidant activity in vitro. These effects do not represent the full potential of its action In vivo. Lavandin oil is widely used in aromatherapy and massages. Despite this, the therapeutic effects of this oil have been largely overlooked in scientific studies when compared to its parent species. When L. x intermedia was studied and compared to L. angustifolia in terms of the biological/therapeutic effects, it usually gave a similar performance, in the case of antimicrobial—even more powerful. The main components of both lavender oil—linalool and linalyl acetate, which is quickly metabolized in vivo to linalool [78]—affect molecular pathways and induce some biological activities. Thus, researchers can expect similar biological activities in general. The differences in composition lie in the secondary and trace constituents, such as camphor, 1,8-cineole, and borneol, which sum up to approximately 30% of the oil. These terpenoids are usually alleged of increased the biocidal action of LI and LL, but increased biocidal properties are a double-edged sword. Of the three mentioned, especially camphor, despite its wide use in pharmacy, is considered to be potentially harmful to health, and the toxicity of camphor is well-documented [79]. Although most cases of camphor poisoning were due to oral ingestion, a few reports indicate that toxic doses of camphor can be absorbed through inhalation and skin contact too. It has been estimated that severe toxicity, which can cause convulsions, may occur in adults at a dose of around 34 milligrams per kilogram of body weight [80]. The typical signs of camphor poisoning when consumed by humans include headaches, nausea, vomiting, dizziness, muscle stimulation causing tremors and twitching, seizures, and delirium. The severity of these symptoms varies depending on the amount of camphor ingested [79]. Therefore, the United States Food and Drug Administration in 1983 set a limit of 11% in consumer products, Ph. Eur. allows the maximum dosage of Camphora racemica or D-Camphora as 10% when admitted topically [79][81]. It is possible that one of the reasons, but nowhere in the literature explicitly stated, why LI and LL are less popular in traditional medicine than lavender is the toxicity of camphor. On the other hand, camphor is appreciated in medicine and commonly used in topical drug formulations. Additionally, lavandin poisoning is uncommon. To the best of the knowledge, there are no described cases of lavandin poisoning in the literature, with the exception of one case involving an 18-month-old infant who ingested a homemade lavandin extract [82]. Unfortunately, the formulation and method of preparation of this extract were not described. However, the authors detected linalyl esters and acetone in both the extract and the patient’s blood, suggesting that it may have been an acetone extract.
Secondary and minor components can influence the overall biological action of linalool. They may be indifferent or interact with each other leading to synergistic or antagonistic effects. Therefore, the activity of essential oil can sometimes be stronger than that of it its main components. It is generally believed that minor chemicals play a critical role in synergistic activity. The compositional complexity and natural variability of the plant material make this kind of research challenging.

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