Membrane Vesicles for Nanoencapsulated Sulforaphane Increased Their Anti-Inflammatory Role on an In Vitro Human Macrophage Model

Abstract

At present, there is a growing interest in finding new non-toxic anti-inflammatory drugs to treat inflammation, which is a key pathology in the development of several diseases with considerable mortality. Sulforaphane (SFN), a bioactive compound derived from Brassica plants, was shown to be promising due to its anti-inflammatory properties and great potential, though its actual clinical use is limited due to its poor stability and bioavailability. In this sense, the use of nanocarriers could solve stability-related problems. In the current study, sulforaphane loaded into membrane vesicles derived from broccoli plants was studied to determine the anti-inflammatory potential in a human-macrophage-like in vitro cell model under both normal and inflammatory conditions. On the one hand, the release of SFN from membrane vesicles was modeled in vitro, and two release phases were stabilized, one faster and the other slower due to the interaction between SFN and membrane proteins, such as aquaporins. Furthermore, the anti-inflammatory action of sulforaphane-loaded membrane vesicles was demonstrated, as a decrease in interleukins crucial for the development of inflammation, such as TNF-α, IL-1β and IL-6, was observed. Furthermore, these results also showed that membrane vesicles by themselves had anti-inflammatory properties, opening the possibility of new lines of research to study these vesicles, not only as carriers but also as active compounds.

Potential of Sulforaphane and Broccoli Membrane Vesicles as Regulators of M1/M2 Human Macrophage Activity

“…Nevertheless, in this experimental model, we found no differences between free SFN and SFN in the encapsulated format, probably due to the powerful effect shown by the free compound. These results are consistent with those obtained previously for TNF-α and IL-6 in macrophage-like cell line models with free SFN [ 35 ] or SFN encapsulated in broccoli membrane vesicles [ 30 ], although the effect recorded was less potent. The effect previously described on IL-1β in cell line models could only be detected when SFN was loaded into broccoli membrane vesicles, achieving a 50% decrease relative to controls cells under inflammatory conditions [ 30 ], whereas the results presented in this study showed that both free and encapsulated SFN completely abrogated the effect of IL-1β production of this cytokine in primary cultures of M1 macrophages.…”

“…These results are consistent with those obtained previously for TNF-α and IL-6 in macrophage-like cell line models with free SFN [ 35 ] or SFN encapsulated in broccoli membrane vesicles [ 30 ], although the effect recorded was less potent. The effect previously described on IL-1β in cell line models could only be detected when SFN was loaded into broccoli membrane vesicles, achieving a 50% decrease relative to controls cells under inflammatory conditions [ 30 ], whereas the results presented in this study showed that both free and encapsulated SFN completely abrogated the effect of IL-1β production of this cytokine in primary cultures of M1 macrophages. These results are also in accordance with previous studies in rats with sciatic endometriosis, where SFN treatment reduced TNF-α, IL-6, and IL-1β.…”

“…Its content in aquaporins, which are intrinsic membrane proteins that can bind and stabilize SFN, contributes to its thermodynamic stability [ 29 ]. Therefore, broccoli membrane vesicles have been used to encapsulate SFN, improving its antiproliferative and anti-inflammatory activity in macrophage model showing two phases of release, one fast occurring through membrane lipids and the other occurring slowly after liberalization with aquaporin proteins [ 30 ]. In addition, membrane vesicles isolated from broccoli can act not only as carriers but also have activity on their own, such as specific bioactivity due to glucosinolates, isothiocyanates, or certain proteins present in the membranes after the isolation procedure [ 30 ].…”

“…The aim of this study was advance in the knowledge previously determined [ 30 ] and analyze the potential clinical applications of broccoli membrane vesicles and SFN, either free or encapsulated, studying the effect on the activity of human M1 and M2 macrophages. Therefore, we established in vitro primary cultures of monocyte-derived macrophages obtained from peripheral blood of healthy donors that were differentiated toward either proinflammatory or M1 or anti-inflammatory or M2 cell profile populations.…”

Plant-derived extracellular vesicles: a novel nanomedicine approach with advantages and challenges

“…P-EVs isolated from broccoli have been found to contain sulforaphane. There have been few reports that suggest it has cancer-protective properties [ 99 ]. P-EVs from grapefruit contain naringenin, fructose, citric acid, glucose, sucrose myo-inositol, quinic acid, oxalic acid, glycolic acids, aucubin in addition to amino acids leucine and isoleucine [ 82 , 91 , 100 ].…”