## 3 Results
### Ability of Caryophyllane sesquiterpenes to Modulate Macrophage Function
A 3-day exposure to 10 \(\upmu\)M \(\beta\)-caryophyllene reduced the levels of proinflammatory cytokines, including TNF-\(\alpha\), PGE-2, and IL-6 in RAW 264.7 cells; in particular, TNF-\(\alpha\) was reduced both with and without LPS, while a lowering in PGE-2 and IL-6 levels occurred in the absence and presence of LPS, respectively [30]. In the same study, \(\beta\)-caryophyllene improved the reduction of the macrophage number induced by metaxalone [30]. Similarly, [PERSON] et al. [31] have found that a short 5 h exposure to a low concentration (5 \(\upmu\)M) of \(\beta\)-caryophyllene counteracted the LPS-induced release of pro-inflammatory cytokines without affecting the basal levels of IL-1 \(\beta\), IL-6, and TNF-\(\alpha\), but lowered them after LPS stimulation. Similarly, a 48 h exposure to \(\alpha\)-humulene, at concentrations of 755 and 1555 \(\upmu\)M, inhibited the inflammatory response induced by LPS, by lowering the release of nitric oxide (NO), in murine macrophages RAW 264.7 [32]. [PERSON] and colleagues studied the modulation of cytokine release induced by LPS in primary splenocytes from BALB/c mice by a 48 h exposure to \(\beta\)-caryophyllene, \(\beta\)-caryophyllene oxide, and \(\alpha\)-humulene (0.25-25 \(\upmu\)M) [33]. In particular, \(\beta\)-caryophyllene was able to lower the levels of both IL-1 and IL-10, which are responsible for pro-inflammatory and anti-inflammatory effects, respectively; moreover, an increase in the T helper cell type 2/1 (Th2/Th1) ratio, associated with anti-inflammatory effects, was found too. Similarly, \(\beta\)-caryophyllene oxide counteracted the LPS-induced inflammation by decreasing the IL-2 levels; by contrast, no effects were highlighted for \(\alpha\)-humulene [33]. Macrophages and monocytes take part in the tumor microenvironment and favor tumor progression through the release of pro-inflammatory cytokines and pro-angiogenic factors; indeed, both tissue-resident macrophages and infiltrated monocytes can give rise to TAM (tumor-associated macrophages), involved in tumor progression [34]. TAM impairs immune the anti-tumor defense and anti-inflammatory response by suppressing the T cells [34] and M2 macrophages [27], while monocyte-derived TAM promotes angiogenesis [34]. Moreover, both the M1 and the M2 macrophages can contribute to cancer development and progression [27]. In this context, [PERSON] et al. [35] evaluated the ability of \(\beta\)-caryophyllene to block the progression of melanoma by affecting macrophage accumulation in tumor tissue (Table 1). To this end, a co-culture model of activated macrophages (M2-M\(\Phi\)s), obtained by murine bone-marrow monocytes exposed to IL-4, mature adipocytes (Mas), and melanoma B16F10s cells was used. Including adipocytes in the cell model is due to the known role of adipose tissue in melanoma progression [35]; indeed, being near the skin, it influences crosstalk between the macrophages and the tumor cells and contributes to tissue inflammation by the release of pro-inflammatory cytokines, as often found in obesity. This disease is also characterized by a chronic and low-grade inflammatory condition, which is supported by a switch to the M1 macrophage phenotype and a recruitment of B and T cells, induced by several factors released by adipose tissue, such as C-C motif chemokine ligand 2 (CCL2) and Toll-like receptor 4 (TLR4) [27; 29]. [PERSON] et al. [35] found that a 24 h exposure to 5 uM \(\beta\)-caryophyllene decreased MCP-1 (monocyte chemoattractant protein-1) and M-CSF (macrophage-colony stimulating factor) levels in the medium of M2-MQs, Mas, and B16F10s, thus suggesting the ability of the compound to lower the macrophage accumulation in tumor tissue and thus limiting the inflammatory conditions that support tumor progression. A reduction in macrophage recruitment by \(\beta\)-caryophyllene was confirmed by the inhibition of their migration when the medium of MAs and B16F10s cells was used as a chemoattractant [35].