Carla Ghelardini

Aims: Chemotherapy‐induced neuropathy is a major side effect of platinum‐based drugs such as oxaliplatin. Given the context‐dependent pro‐inflammatory or neuroprotective actions of interferon‐γ (IFN‐γ), this study investigated its role in an in vitro model of oxaliplatin‐induced neurotoxicity (OIPN). Methods: Rat organotypic spinal cord slices were exposed to oxaliplatin to model chemotherapy‐induced neurotoxicity responses in vitro. IFN‐γ cytotoxicity was assessed at 6 and 24 h. Neurotoxicity, inflammatory mediators, neurotrophic factors, and markers of glial and neuronal activation were analyzed by biochemical and immunohistochemical assays. Connexin 43 (Cx43) was assessed using Gap19 and Gap27 to probe hemichannel and gap‐junction contributions. Electrophysiological recordings characterized neuronal excitability. Results: IFN‐γ (6.5–200 ng/mL) was non‐toxic and, at 65 ng/mL, significantly reduced oxaliplatin‐induced neurotoxicity, decreasing LDH release and suppressing pro‐inflammatory mediators (CCL2, SerpinA3N, CGRP, substance P, c‐Fos). IFN‐γ modulated its signaling by reducing IFNγR and increasing IRF1 expression. It prevented oxaliplatin‐induced overexpression of neurotrophic factors and reduced glial and neuronal hyperactivation markers, preserving neuronal integrity. IFN‐γ reduced Cx43 expression and hemichannel activity, and Gap19 reproduced selected protective effects. Electrophysiology showed attenuation of oxaliplatin‐induced neuronal hyperexcitability. Conclusion: IFN‐γ attenuates oxaliplatin‐induced neurotoxic responses in an in vitro spinal cord model. The parallel reduction in Cx43 expression and hemichannel activity supports further investigation of Cx43 signaling as a candidate pharmacological target in OIPN.

Visceral pain is a major symptom of inflammatory bowel diseases (IBDs), requiring effective treatment strategies. Gut epithelium, beyond maintaining barrier integrity and microbiota homeostasis, modulates neurosensorial circuitries, influencing visceral sensitivity. Moringa oleifera constituents show beneficial properties that may counteract inflammation‐induced epithelial dysfunction and visceral hypersensitivity. Among these, we investigated moringin (MOR), the isothiocyanate derived from the myrosinase (MYR)‐mediated hydrolysis of glucomoringin (GMG). A three‐stage experimental strategy was applied. First, murine colon organoids were exposed to a pro‐inflammatory cytokine cocktail (CKs; TNF‐α, IL‐1β, IL‐6; 10 ng mL ⁻¹ , 6 h) to model epithelial inflammatory stress and treated with MOR (2–30 μM). Second, epithelial–neuronal communication was assessed by exposing primary dorsal root ganglion (DRG) neurons to conditioned media from inflamed organoids, with (CM ^{CKs + MOR 30 μM} ) or without (CM ^CKs ) MOR treatment. Third, the therapeutic relevance of these findings was validated in vivo using a dextran sulfate sodium (DSS)–induced colitis model, in which mice received oral administration of MYR‐bioactivated GMG (GMG + MYR) at different doses (30–100 mg kg ⁻¹ ), followed by behavioral and histological assessments. CKs‐treated organoids showed cytotoxicity, increased superoxide dismutase activity, and upregulation of Ccl2 , Ccnd1 , Mki67, and Pyy . MOR 30 μM co‐treatment normalized oxidative stress and gene expression, although cytotoxicity remained unaffected. CM ^CKs increased c‐Fos and CGRP expression in DRG neurons, while CM ^{CKs + MOR 30 μM} prevented this response, indicating MOR modulation of epithelial inflammation‐driven neuronal activation. In DSS‐treated mice, oral GMG (30–100 mg kg ⁻¹ ) + MYR dose‐dependently reduced visceral hypersensitivity, pain‐associated behavioral alterations, and colonic damage. Overall, MOR/GMG + MYR showed gut‐protective and analgesic effects under intestinal inflammation. Intestinal organoids effectively model inflammation and therapeutic responses, potentially reducing animal use in IBD research.

Chronic pain represents a complex debilitating condition that extends beyond the protective function of physiological pain, often persisting as an independent disease entity. Chronic primary and secondary pain syndromes reflect a multifaceted continuum involving nociceptive, neuropathic and nociplastic mechanisms. The maladaptive plasticity of the peripheral and central nervous system (encompassing the ascending and descending pain pathways) sustains hypersensitivity and correlates with comorbid alterations in mood, cognition, sleep, and fatigue, underpinned by functional reorganization of brain networks. In this scenario traditional analgesics frequently demonstrate limited efficacy, while current guidelines recommend antiepileptic agents and antidepressants, particularly gabapentinoids and duloxetine, a first line pharmacological options. This review explores the mechanistic rationale and clinical evidence supporting the combined use of gabapentinoids and duloxetine in chronic pain management. These agents act on distinct yet complementary targets: gabapentinoids reduce excitatory neurotransmission via modulation of calcium channel activity, while duloxetine restores descending noradrenergic inhibition and alleviates comorbid symptoms. Clinical trial and meta-analyses highlight their individual efficacy in diabetic peripheral neuropathy, postherpetic neuralgia, and fibromyalgia. Among gabapentinoids, pregabalin exhibits a favorable pharmacokinetic profile that allows rapid titration and demonstrates effectiveness against anxiety-related sleep disorders. Importantly, emerging evidence suggests that their combination may yield superior pain relief and functional improvement compared with monotherapy, particularly in patients with residual pain. This review provides a proof of concept by bridging theoretical knowledge and real-life clinical settings aiming to develop treatment protocols based on predominant pain mechanisms that can effectively control hypersensitivity and improve quality of life.

Chronic gastrointestinal pain is a hallmark of most intestinal pathologies, yet effective treatments remain elusive given the complexity of the underlying mechanisms. Aiming to investigate the intestinal epithelium contribution to visceral pain modulation in dysbiosis context, we first demonstrated that intracolonic instillation of microbe-free fecal supernatants from mice with post-inflammatory dysbiosis induced by dextran sodium sulfate (FS^DSS) provokes visceral hypersensitivity in recipient mice. Epithelium involvement in the response to FS^DSS was analyzed through a novel in vitro approach comprising murine epithelial colon organoids and primary dorsal root ganglia (DRG) neurons. FS^DSS treatment induced growth and metabolic impairment in colon organoids, which revealed a dysbiosis-driven epithelial dysfunction. Notably, the combination of FS^DSS and conditioned medium from FS^DSS-treated colon organoids induced an increase in DRG neuron intrinsic excitability, along with greater immunoreactivity to c-Fos and calcitonin-gene related peptide, implicating an integrated role of both microbial and epithelial products in visceral sensitivity regulation. By investigating the underlying signaling, metabolomic analysis revealed reduced levels of short chain fatty acids in FS^DSS, such as butyrate, acetate, valerate, and propionate. Moreover, transcriptomic analysis of FS^DSS-treated colon organoids showed the dysregulated expression of several signaling factors by which intestinal epithelium may modulate sensory neuron excitability, including proteases, cytokines, neuromodulators, growth factors, and hormones. These findings provide novel insights into the role of gut epithelium in the modulation of sensory neuron excitability under dysbiosis conditions, emphasizing that targeting epithelial-neuronal signaling might represent a promising therapeutic strategy for visceral pain management.

Background: Chemotherapy-induced neuropathic pain is a major side effect of antineoplastic treatment. This study investigated the neuroprotective potential of Acmella oleracea L. extracts containing the N-alkylamide spilanthol, phenolic acids, and glycosylated flavonoids. Methods: Hydroalcoholic extracts of aerial parts (AP) and roots (R) of in vitro seedlings were quali-quantitatively characterized by HPLC-DAD-MS and by ¹H-NMR. Different concentrations (15–150 µg/mL) of AP and R were tested in SH-SY5Y cells differentiated into neurons exposed to oxaliplatin (10 µM), assessing cell viability (MTT), cytotoxicity (LDH), SOD activity, and expression of ATF-3, Ire1α, and Nf-H genes. To evaluate the impact on neuropathic pain, CD-1 mice were treated intraperitoneally with oxaliplatin (2.4 mg/kg), the effect of AP and R extracts (200–1200 mg/kg) were measured by the cold plate test. Results: AP extract was rich in phenols and alkylamides, whereas R extract showed higher phenolic levels but lower alkylamides content. Both extracts significantly reduced mortality and cytotoxicity and counteracted oxidative imbalance by enhancing SOD activity. Gene expression analysis confirmed their neuroprotective effects. In vivo, oxaliplatin induced a 50% reduction in pain threshold, while acute treatment with AP and R extracts dose-dependently alleviated neuropathic pain. Despite the lower spilanthol content in R extract, its efficacy was comparable to AP, suggesting an important role of phenolic compounds. Conclusions: Extracts from both aerial parts and roots of A. oleracea show promise in alleviating chemotherapy-induced neuropathy through mechanisms not solely related to spilanthol. Further studies to elucidate the contribution of phenolic components are desirable.

Octocorals have proven to be a prolific source of bioactive natural products, exhibiting a wide spectrum of pharmacological activities. Among octocorals, Pennatulaceans, commonly known as sea pens, are among the most dominant soft coral species living in benthic communities. Nonetheless, reports on bioactivity and chemical investigations of this genus are scarce. This prompted us to shed light on the pharmacological potential of the extracts of the sea pen Pennatula phosphorea, Linneus 1758, and gain an overview of its metabolome. Crude octocoral extracts, obtained with a modified Kupchan extraction protocol, were assessed for their bioactivity potential, revealing the hexanic extract to exert anti-inflammatory effects and interesting protective properties in an in vitro model of sarcopenia and in auditory HEI-OC1 cisplatin-treated cells, while the chloroformic extract was active in reducing A375 melanoma cell viability in a concentration-dependent manner. An untargeted metabolomic analysis unveiled that P. phosphorea collects a wide array of glycerophospholipids and phosphosphingolipids belonging to the ceramide phosphoinositol class, which were exclusive or more abundant in the hexanic extract. Their proven anti-inflammatory and cytoprotective effects could demonstrate the activity shown by the P. phosphorea hexanic extract. In addition, a group of prostaglandins, eluted mainly in the chloroformic extract, were putatively annotated. Since prostanoids from marine origin have been demonstrated to exert cytotoxic and anti-proliferative properties against various cancer cell lines, the presence of PGs in the P. phosphorea chloroform extract could justify its anti-melanoma activity. This is the first report on the presence of glycerophospholipids, phosphosphingolipids, and prostaglandins, along with the identification of novel congeners, in sea pens.