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Multiple myeloma is a plasma cell neoplasm characterized by the production of unfolded immunoglobulins, which cause endoplasmic reticulum (ER) stress and sensitivity to proteasome inhibition. The genomic landscape of multiple myeloma is characterized by the loss of several genes rarely mutated in other cancers that may underline specific weaknesses of multiple myeloma cells. One of these is FAM46C that is lost in more than 10% of patients with multiple myeloma. We show here that FAM46C is part of a new complex containing the ER-associated protein FNDC3A, which regulates trafficking and secretion and, by impairing autophagy, exacerbates proteostatic stress. Reconstitution of FAM46C in multiple myeloma cells that had lost it induced apoptosis and ER stress. Apoptosis was preceded by an increase of intracellular aggregates, which was not linked to increased translation of IgG mRNA, but rather to impairment of autophagy. Biochemical analysis showed that FAM46C requires interaction with ER bound protein FNDC3A to reside in the cytoplasmic side of the ER. FNDC3A was lost in some multiple myeloma cell lines. Importantly, depletion of FNDC3A increased the fitness of FAM46C-expressing cells and expression of FNDC3A in cells that had lost it recapitulated the effects of FAM46C, inducing aggregates and apoptosis. FAM46C and FNDC3A formed a complex that modulates secretion routes, increasing lysosome exocytosis. The cellular landscape generated by FAM46C/FNDC3A expression predicted sensitivity to sphingosine kinase inhibition. These results suggest that multiple myeloma cells remodel their trafficking machinery to cope with ER stress.Significance:This study identifies a new multiple myeloma–specific tumor suppressor complex that regulates autophagy and unconventional secretion, highlighting the sensitivity of multiple myeloma cells to the accumulation of protein aggregates.

Supplementary Figures

Secretomics analysis with/without wtFAM46C

The Mediterranean diet (MD) is becoming a milestone for the prevention of chronic diseases, such as cardiovascular diseases (CVDs), Alzheimer’s and Parkinson’s disease. Ancel Keys in the 1950’s showed a low mortality rate, particularly for coronary heart disease, among people resident in the Mediterranean area. The MD is characterized by the intake of the high amount of vegetables, fruit, and cereals and regular but moderate consumption of wine, fish, and dairy products, while olive oil is the main source of culinary fat. Therefore, it is principally a plant-based diet rich in polyphenols, a heterogeneous category of compounds with different properties and bioavailabilities. Among polyphenols, anthocyanins have been combined into the human food regime for centuries. They have been utilized as traditional herbal remedies for their ability to treat several conditions, as potent anti-oxidants, anti-diabetic and anti-carcinogenic compounds. This review summarizes our knowledge on the health-enhancing component of the anthocyanins-rich diet.

Unfolded protein response (UPR) is an adaptive response which is used for re-establishing protein homeostasis, and it is triggered by endoplasmic reticulum (ER) stress. Specific ER proteins mediate UPR activation, after dissociation from chaperone Glucose-Regulated Protein 78 (GRP78). UPR can decrease ER stress, producing an ER adaptive response, block UPR if ER homeostasis is restored, or regulate apoptosis. Some tumour types are linked to ER protein folding machinery disturbance, highlighting how UPR plays a pivotal role in cancer cells to keep malignancy and drug resistance. In this review, we focus on some molecules that have been revealed to target ER stress demonstrating as UPR could be a new target in cancer treatment.

The present study was conducted at Surgical Pathology Department of “Santo Spirito” hospital in Casale Monferrato (AL) and in collaboration with the Cellular Biology and Physiology laboratory located in the institution of DiSIT-Università del Piemonte Orientale. In the pathology facility a retrospective analysis on the immunohistochemical expression of CD10 marker in malignant pleural mesothelioma was delineated. For this purpose it was collected a sample of 63 pleural biopsies with a diagnosis of pleural mesothelioma come in the period of January 2015 and November 2020. In this cohort, CD10 expression was assessed from a prognostic perspective and in relation to morphologic aspects of the neoplasm. Results from previous analysis built the basis of the experiment conducted in the DiSIT facility. Two pleural mesothelioma cell lines, REN and MM98, got incubated with the cytokine TGF-β aiming to analyze the potential relationship between epithelial-to-mesenchymal transition (EMT) and CD10 cell expression pattern. Bioinformatic server Haddock 2.4 helped to predict and model potential cross-talk between EMT underlying pathways and CD10 signalling. Objectives: The present study aimed to assess prognostic role of CD10 immunohistochemical expression in a cohort of patients affected by pleural mesothelioma. Second goal was to analyze the relationship between CD10 biopsy expression, phenotypical and imunophenotypical features (referring in particular to PDL-1 expression) of the neoplasm. Results from this retrospective analysis established next aim of the study: induction of EMT in two mesothelioma cell lines, REN and MM98, and analysis of change in CD10 cellular pattern. Final goal was to predict a potential cross-talk between EMT related pathways and CD10 activity based on bioinformatic server Haddock 2.4.Methods: Immunohistochemical analysis of biopsies and cell cultures were processed on Ventana Benchmark immunostainer. Tools adopted in qRT-PCR of cell cultures were: Power Sybr Green Mastermix probes (Ambion Austin, TX, USA) and KiCqStart® SYBR® Green primers (Sigma-Aldrich); Real-Time PCR CFX384 detection system (Bio-Rad Laboratories, Hercules, CA, USA). Gene expressione was measured with ∆∆Ct method. Parametrization of CD10 immunohistochemical expression was accomplished with cell imaging software CellProfiler 3.1.9. Statistical analysis were ran with R software version 3.6.2. Results: CD10 showed to be a negative prognostic factor, positively associated to neoplastic grade, in the cohort of patients analyzed. Induction of EMT in REN and MM98 TGF-β treated cultures, augmented CD10 cytosolic expression against non-treated cultures. Specifically, a change in CD10 immunoexpression pattern, from membranous to cytosolic, was observed for EMT+ REN cell culture. This data agrees with prevalent CD10 cytosolic expression observed in mesothelioma with low differentiation, where differentiation represents a function of EMT. Furthermore, Haddock data highlighted a potential promotion of EMT by CD10 via a NF-kB interaction. CD10 could establish a positive feedback loop with EMT. Conclusion: CD10 represents a positive marker of neoplastic progression in pleural mesothelioma that, based on its expression level, can stratify survival. Experimental data showed that CD10 could be upregulated by EMT pathways. Bionformatic analysis also suggest that CD10 could interact with EMT establishing a positive feedback loop. Clarifying tumorigenicity of CD10 in mesothelioma is fundamental to define its possible therapeutic role.

Cancer still remains a leading cause of death despite improvements in diagnosis, drug discovery and therapy approach. Therefore, there is a strong need to improve methodologies as well as to increase the number of approaches available. Natural compounds of different origins (i.e., from fungi, plants, microbes, etc.) represent an interesting approach for fighting cancer. In particular, synergistic strategies may represent an intriguing approach, combining natural compounds with classic chemotherapeutic drugs to increase therapeutic efficacy and lower the required drug concentrations. In this review, we focus primarily on those natural compounds utilized in synergistic approached to treating cancer, with particular attention to those compounds that have gained the most research interest.

Malignant mesothelioma is a rare cancer arising from the serosal surfaces of the body, mainly from the pleural layer. This cancer is strongly related to asbestos exposure and shows a very inauspicious prognosis, because there are scarce therapeutic options for this rare disease. Thus, there is an urgent need to develop novel therapeutic approaches to treat this form of cancer. To explore the biology of malignant pleural mesothelioma (MPM), we previously observed that MPM cell lines show high expression of the GRP78 protein, which is a chaperone protein and the master regulator of the unfolded protein response (UPR) that resides in the endoplasmic reticulum (ER). Based on our previous studies showing the importance of GRP78 in MPM, we observed that BOLD-100, a specific modulator of GRP78 and the UPR, shows cytotoxicity against MPM cells. Our studies demonstrated that BOLD-100 increases ROS production and Ca2+ release from the ER, leading to ER stress activation and, ultimately, to cell death. Our in vitro data strongly suggest that BOLD-100 inhibits the growth of MPM cell lines, proposing the application as a single agent, or in combination with other standard-of-care drugs, to treat MPM.

Stem cells present in the epidermis, and hair follicle, guarantee the conservation of adult skin maintenance and hair renewal, but they also play a pivotal role in wound repair and tissue regeneration. Adult stem cells present in the epidermis are also responsible for epidermis different layers' regeneration.We here summarize the epidermal stem cells information in term of their central features in stem cells niche, their signalling pathways and their maintenance, and activation.

Wound repair is a dynamic process during which crucial signaling pathways are regulated by growth factors and cytokines released by several kinds of cells directly involved in the healing process. However, the limited applications and heterogeneous clinical results of single growth factors in wound healing encouraged the use of a mixture of bioactive molecules such as platelet derivatives for best results in wound repair. An interesting platelet derivative, obtained from blood samples, is platelet lysate (PL), which has shown potential clinical application. PL is obtained from freezing and thawing of platelet-enriched blood samples. Intracellular calcium (Ca2+) signals play a central role in the control of endothelial cell survival, proliferation, motility, and differentiation. We investigated the role of Ca2+ signaling in the PL-driven endothelial healing process. In our experiments, the functional significance of Ca2+ signaling machinery was highlighted performing the scratch wound assay in presence of different inhibitors or specific RNAi. We also pointed out that the PL-induced generation of intracellular ROS (reactive oxygen species) via NOX4 (NADPH oxidase 4) is necessary for the activation of TRPM2 and the resulting Ca2+ entry from the extracellular space. This is the first report of the mechanism of wound repair in an endothelial cell model boosted by the PL-induced regulation of [Ca2+]i.

The preparation of smart Ln:ZrO2@SiO2 nanoplatforms with grafted photosensitizer (Rose Bengal) which couple optical imaging with photo-dynamic therapy (PDT) is presented. A careful control of the lanthanide dopant loading is considered to enhance the photoemission properties of the lanthanide ions (Er, Pr, Yb) inside the ZrO2 crystal structure. The nanosystem with the lowest lanthanide loading maintains the size, phase and morphology of pristine ZrO2 nanoparticles and exhibit the best performances in term of the overall luminescence properties. Upon functionalization with a silica shell to covalently bound Rose Bengal, a theranostic platform is prepared which is very efficient in singlet oxygen generation, as demonstrated by EPR and UV–vis spectroscopy studies. Preliminary cell viability tests show that while both pristine and Ln doped ZrO2 nanoparticles do not exert cytotoxicity, neither upon illumination nor in dark condition, Rose Bengal grafted samples are able to significantly reduce cell viability under light exposure, thus confirming the high potential of these nanoparticles as PDT tools.

Stem cells present in the epidermis, and hair follicle, guarantee the conservation of adult skin maintenance and hair renewal, but they also play a pivotal role in wound repair and tissue regeneration. Adult stem cells present in the epidermis are also responsible for epidermis different layers' regeneration.We here summarize the epidermal stem cells information in term of their central features in stem cells niche, their signalling pathways and their maintenance, and activation.

Multiple myeloma is a plasma cell neoplasm characterized by the production of unfolded immunoglobulins, which cause endoplasmic reticulum (ER) stress and sensitivity to proteasome inhibition. The genomic landscape of multiple myeloma is characterized by the loss of several genes rarely mutated in other cancers that may underline specific weaknesses of multiple myeloma cells. One of these is FAM46C that is lost in more than 10% of patients with multiple myeloma. We show here that FAM46C is part of a new complex containing the ER-associated protein FNDC3A, which regulates trafficking and secretion and, by impairing autophagy, exacerbates proteostatic stress. Reconstitution of FAM46C in multiple myeloma cells that had lost it induced apoptosis and ER stress. Apoptosis was preceded by an increase of intracellular aggregates, which was not linked to increased translation of IgG mRNA, but rather to impairment of autophagy. Biochemical analysis showed that FAM46C requires interaction with ER bound protein FNDC3A to reside in the cytoplasmic side of the ER. FNDC3A was lost in some multiple myeloma cell lines. Importantly, depletion of FNDC3A increased the fitness of FAM46C-expressing cells and expression of FNDC3A in cells that had lost it recapitulated the effects of FAM46C, inducing aggregates and apoptosis. FAM46C and FNDC3A formed a complex that modulates secretion routes, increasing lysosome exocytosis. The cellular landscape generated by FAM46C/FNDC3A expression predicted sensitivity to sphingosine kinase inhibition. These results suggest that multiple myeloma cells remodel their trafficking machinery to cope with ER stress. Significance: This study identifies a new multiple myeloma–specific tumor suppressor complex that regulates autophagy and unconventional secretion, highlighting the sensitivity of multiple myeloma cells to the accumulation of protein aggregates.

The neurotransmitter glutamate increases cerebral blood flow by activating postsynaptic neurons and presynaptic glial cells within the neurovascular unit. Glutamate does so by causing an increase in intracellular Ca2+ concentration ([Ca2+ ]i ) in the target cells, which activates the Ca2+ /Calmodulin-dependent nitric oxide (NO) synthase to release NO. It is unclear whether brain endothelial cells also sense glutamate through an elevation in [Ca2+ ]i and NO production. The current study assessed whether and how glutamate drives Ca2+ -dependent NO release in bEND5 cells, an established model of brain endothelial cells. We found that glutamate induced a dose-dependent oscillatory increase in [Ca2+ ]i , which was maximally activated at 200 μM and inhibited by α-methyl-4-carboxyphenylglycine, a selective blocker of Group 1 metabotropic glutamate receptors. Glutamate-induced intracellular Ca2+ oscillations were triggered by rhythmic endogenous Ca2+ mobilization and maintained over time by extracellular Ca2+ entry. Pharmacological manipulation revealed that glutamate-induced endogenous Ca2+ release was mediated by InsP3 -sensitive receptors and nicotinic acid adenine dinucleotide phosphate (NAADP) gated two-pore channel 1. Constitutive store-operated Ca2+ entry mediated Ca2+ entry during ongoing Ca2+ oscillations. Finally, glutamate evoked a robust, although delayed increase in NO levels, which was blocked by pharmacologically inhibition of the accompanying intracellular Ca2+ signals. Of note, glutamate induced Ca2+ -dependent NO release also in hCMEC/D3 cells, an established model of human brain microvascular endothelial cells. This investigation demonstrates for the first time that metabotropic glutamate-induced intracellular Ca2+ oscillations and NO release have the potential to impact on neurovascular coupling in the brain.

Candida glabrata is an opportunistic pathogen, associated with endocarditis, meningitis, and disseminated disease, and also with complicated vaginitis. Essential oils derived from aromatic plants are known in traditional medicine as antimicrobial agents and have antifungal properties. The aim of this work was to evaluate whether 12 tested essential oils (tea tree, laurel, anise, basil, bergamot, lavender, mint, oregano, grapefruit, rosemary, winter savory, and ginger) could have a transverse effect on C. glabrata sensitive strains but above all on strains resistant to the three main azole antifungals used (clotrimazole, fluconazole, itraconazole). For this reason, different strains of C. glabrata, vaginal isolated, were characterized (disk diffusion assay, minimal inhibitory concentration) with respect to their response to such antifungals. Electron microscopy analyses were performed to examine cellular damages in depth. Subsequently, we wanted to evaluate the effect of the oils on human cells to estimate their potential cytotoxicity. Oregano and winter savory were the two most effective essential oils, inducing growth inhibition, cell damage of C. glabrata strains (both sensitive and resistant to azole antifungal drugs), and medium–high level of toxicity against human keratinocytes. The results of this work support the research for new alternatives or complementary therapies against vaginal candidiasis.

Silk fibroin may be chemically modified by grafting, with the purpose of improving its properties according to the desired function. In this study, silk fabrics from Bombyx mori silk fibres were grafted with 2-hydroxyethyl methacrylate (HEMA), as well as a binary mixture of HEMA and 4-hydroxybutyl acrylate (HBA). The samples were then electrospun from trifluoroacetic acid and treated with aqueous methanol. The% weight gains ascribable to HEMA and HBA were successfully determined through Raman spectroscopy. PolyHEMA made the fibres more hydrophilic and hindered crystallization into β-sheet only upon electrospinning and treatment with aqueous methanol; the presence of the HBA component in the grafting mixture did not further decrease the ability of silk fibroin to rearrange into β-sheet, due to its low contents (below 5%) under the used experimental conditions. Fibrillation partially occurred in the grafted fabrics; the electrospun samples maintained their nanostructured morphology. The surface of the substrates under investigation was compatible with cell attachment and growth, which were higher for the nanofibres. Cell adhesion and proliferation may be modulated by varying the surface chemistry and topography of the fabrics; grafting improved the surface properties of silk fibroin for enhanced functional performance in view of biomedical applications.

Microwave (MW) thermal heating has been proposed as an efficient method for honey liquefaction, while maintaining honey quality criteria. However, little is known about the effects of MW thermal heating on honey antibacterial activity. In this study, we aimed to determine the effects of MW heating on the antibacterial activity of raw rapeseed honeys against Pseudomonas aeruginosa and Staphylococcus aureus, with a particular focus on two major bee-derived antibacterial components, defensin-1 and hydrogen peroxide (H2O2). Our results demonstrated that MW thermal heating completely abolished honey antibacterial activity whereas conventional thermal treatment at 45 and 55°C did not affect the antibacterial activity of honey samples. A significant decrease in both glucose oxidase activity and H2O2 production as well as defensin-1 amount was observed in MW-treated samples. Given that defensin-1 and H2O2 are regular antibacterial components of all honeys, MW heating may have similar negative effects on every type of crystallized/liquid honey.

Royal jelly (RJ) has successfully been used as a remedy in wound healing. RJ has multiple effects, including antibacterial, anti-inflammatory and immunomodulatory activities, in various cell types. However, no component(s) (other than antibacterial) have been identified in RJ-accelerated wound healing. In this study, we demonstrate that keratinocytes are responsible for the elevated production of matrix metalloproteinase-9 (MMP-9) after incubation with a water extract of RJ. Furthermore, the keratinocyte migration and wound closure rates were significantly increased in the presence of RJ extract. MMP-9 production was reduced significantly following proteinase K treatment but remained stable after heat treatment, indicating that active component(s) have a proteinous character. To identify the component responsible for inducing MMP-9 production, RJ extract was fractionated using C18 RP-HPLC. In fractions exhibiting stimulatory activity, we immunochemically detected the bee-derived antibacterial peptide, defensin-1. Defensin-1 was cloned, and recombinant peptide was produced in a baculoviral expression system. Defensin-1 stimulated MMP-9 secretion from keratinocytes and increased keratinocyte migration and wound closure in vitro. In addition, defensin-1 promoted re-epithelisation and wound closure in uninfected excision wounds. These data indisputably demonstrate that defensin-1, a regular but concentration variable factor found in honey and RJ, contributes to cutaneous wound closure by enhancing keratinocyte migration and MMP-9 secretion.

Honey is a natural product well known by humankind and now reconsidered for its use as topical agent for wound and burn treatments. Floral honey is made by honeybees from the nectar of blossoms, while honeydew honey is prepared from secretions of plants or excretions of plant-sucking insects. Chemical composition is different between blossom and honeydew honeys and there is very few information about the biological properties of honeydew honey. So, this study was specifically designed to explore the potential wound healing effects of the honeydew honey. We used in vitro scratch wound healing model consisting of fibroblasts and keratinocytes. Data showed that honeydew honeys is able to increase wound closure by acting both on fibroblasts and keratinocytes. Based on our findings, honeydew honey has the potential to be useful for clinical settings.

Cell migration is a crucial step for wound healing. Assays able to evaluate cell migration are very useful to evaluate in vitro wound healing. Scratch wound assay creates a gap in confluent monolayer of keratinocytes to mimic a wound. The protocol of scratch wound is based on few steps: cell culture preparation, scratch wound assay, data acquisition, and data analysis.

Poly(L-lactic) acid (PLLA) has been widely employed in tissue engineering due to its mechanical properties, biodegradability and biocompatibility. The layer-by-layer (LbL) technique was here proposed as a simple method to impart bioactivity to the surface of PLLA substrates. Aminolysis treatment was applied to introduce amino groups on the surface of PLLA solvent cast films. Then, PLLA films were coated with heparin (HE)/chitosan (CH) multilayer by the LbL technique. Each functionalization step was characterized through physico-chemical and morphological analyses. Aminolysis treatment increased film surface wettability (64.8° ± 2.4° against 74.6° ± 1.3° for untreated PLLA) due to the formation of surface amino groups, which were quantified by acid orange colorimetric assay (0.05 nmol/mm

Since Biblical times, honey has been utilized in &ldquo, folk medicine&rdquo, and in recent decades the positive qualities of honey have been re-discovered and are gaining acceptance. Scientific literature states that honey has been successfully utilized on infections not responding to classic antiseptic and antibiotic therapy, because of its intrinsic H2O2 production. In our study, we demonstrated the involvement of H2O2 as a main mediator of honey regenerative effects on an immortalized human keratinocyte cell line. We observed that this extracellularly released H2O2 could pass across the plasma membrane through a specific aquaporin (i.e., AQP3). Once in the cytoplasm H2O2, in turn, induces the entry of extracellular Ca2+ through Melastatin Transient Receptor Potential 2 (TRPM2) and Orai1 channels. Honey-induced extracellular Ca2+ entry results in wound healing, which is consistent with the role played by Ca2+ signaling in tissue regeneration. This is the first report showing that honey exposure increases intracellular Ca2+ concentration ([Ca2+]i), due to H2O2 production and redox regulation of Ca2+-permeable ion channels, opening up a new horizon for the utilization of the honey as a beneficial tool.

Cell migration is a crucial step for wound healing. Assays able to evaluate cell migration are very useful to evaluate in vitro wound healing. Scratch wound assay creates a gap in confluent monolayer of keratinocytes to mimic a wound. The protocol of scratch wound is based on few steps: cell culture preparation, scratch wound assay, data acquisition, and data analysis.

High mobility group box protein-1 (HMGB1) is released from cells under various pathological conditions and it plays a pivotal role as an alarmin signaling tissue damage. Little is known about the impact of HMGB1 in bone repair and remodeling. To this aim, we focused on HMGB1-induced effects on the in vitro osteoblast model SaOS-2. Cell proliferation was stimulated with a maximum at concentration of 2.5 nM, and such a dose also stimulated cell migration and scratch wound healing. We then characterized the modulatory effect of HMGB1 on bone biology, by using osteogenesis/mineralization assays, a PCR array, and the analysis of a series of osteogenic markers. We performed also a proteomic screening using SWATH-MS on SaOS-2 cell exposed to HMGB1 and we provide evidence for proteins modulated in HMGB1 exposed cells. Taken together, our data demonstrate that SaOS-2 cell proliferation, migration, and osteogenic differentiation were increased by HMGB1. We, therefore, propose that HMGB1 could be a potent bone-remodeling signal but the physiological meaning of this property remains to be more ascertained. J. Cell. Biochem. 117: 2559-2569, 2016. © 2016 Wiley Periodicals, Inc.

Poloxamer P407 (P407) is a Food and Drug Administration approved triblock copolymer; its hydrogels show fast dissolution in aqueous environment and weak mechanical strength, limiting their in vivo application. In this work, an amphiphilic poly(ether urethane) (NHP407) was synthesized from P407, an aliphatic diisocyanate (1,6-hexanediisocyanate) and an amino acid derived diol (N-Boc serinol). NHP407 solutions in water-based media were able to form biocompatible injectable thermosensitive hydrogels with a lower critical gelation temperature behavior, having lower critical gelation concentration (6% w/v versus 18% w/v), superior gel strength (G′ at 37 °C about 40 000 Pa versus 10 000 Pa), faster gelation kinetics (

Oxidative polymerization of tyrosol by horseradish peroxidase (HRP)-H2O2 afforded an insoluble product (oligotyrosol, OligoTyr) consisting of mixture of linear oligomers (up to 11-mer) with limited benzylic branching points, as evidenced by ESI-MS and solid state 13C NMR analysis. OligoTyr proved to be significantly more active than tyrosol in several antioxidant assays and was not toxic to human osteosarcoma SaOS-2 cells, stimulating alkaline phosphatase (ALP) activity at day 7 in a similar manner as tyrosol. However, when loaded at 5% w/w into highly porous polylactic acid (PLA) scaffolds featuring hierarchical structures, OligoTyr caused a significant increase in the ALP activity of SaOS-2 cells compared to PLA alone, while tyrosol was completely inactive. A release of ca. 5% from PLA was determined after 1 week in a physiological medium. No significant influence on calcium release from PLA scaffolds containing 5% β-tricalcium phosphate was observed.

Honey is a natural product with a long use in traditional medicine and is well recognized to regulate different biological events. It is an important source of various biological or pharmacological molecules and, therefore, there is a strong interest to explore their properties. Evidence is growing that honey may have the potential to be an anticancer agent acting through several mechanisms. Here we observed for the first time in a cancer cell line a possible mechanism through which honey could induce an alteration in the intracellular reactive oxygen species and homeostatic balance of intracellular calcium concentration leading to cell death by apoptosis. This mechanism seems to be enhanced by manuka honey&rsquo, s ability to maintain high H2O2 permeability through aquaporin-3.

Since Biblical times, honey has been utilized in “folk medicine”, and in recent decades the positive qualities of honey have been re-discovered and are gaining acceptance. Scientific literature states that honey has been successfully utilized on infections not responding to classic antiseptic and antibiotic therapy, because of its intrinsic H2O2 production. In our study, we demonstrated the involvement of H2O2 as a main mediator of honey regenerative effects on an immortalized human keratinocyte cell line. We observed that this extracellularly released H2O2 could pass across the plasma membrane through a specific aquaporin (i.e., AQP3). Once in the cytoplasm H2O2, in turn, induces the entry of extracellular Ca2+ through Melastatin Transient Receptor Potential 2 (TRPM2) and Orai1 channels. Honey-induced extracellular Ca2+ entry results in wound healing, which is consistent with the role played by Ca2+ signaling in tissue regeneration. This is the first report showing that honey exposure increases intracellular Ca2+ concentration ([Ca2+]i), due to H2O2 production and redox regulation of Ca2+-permeable ion channels, opening up a new horizon for the utilization of the honey as a beneficial tool.

Cancer therapy often relies on the combined action of different molecules to overcome drug resistance and enhance patient outcome. Combined strategies relying on molecules with different pharmacokinetics often fail due to the lack of concomitant tumor accumulation and, thus, to the loss of synergistic effect. Due to their ability to enhance treatment efficiency, improve drug pharmacokinetics, and reduce adverse effects, polymer nanoparticles (PNPs) have been widely investigated as co-delivery vehicles for cancer therapies. However, co-encapsulation of different drugs and probes in PNPs requires a flexible polymer platform and a tailored particle design, in which both the bulk and surface properties of the carriers are carefully controlled. In this work, we propose a core-shell PNP design based on a polyurethane (PUR) core and a phospholipid external surface. The modulation of the hydrophilic/hydrophobic balance of the PUR core enhanced the encapsulation of two chemotherapeutics with dramatically different water solubility (Doxorubicin hydrochloride, DOXO and Docetaxel, DCTXL) and of Iron Oxide Nanoparticles for MRI imaging. The outer shell remained unchanged among the platforms, resulting in un-modified cellular uptake and in vivo biodistribution. We demonstrate that the choice of PUR core allowed a high entrapment efficiency of all drugs, superior or comparable to previously reported results, and that higher core hydrophilicity enhances the loading efficiency of the hydrophilic DOXO and the MRI contrast effect. Moreover, we show that changing the PUR core did not alter the surface properties of the carriers, since all particles showed a similar behavior in terms of cell internalization and in vivo biodistribution. We also show that PUR PNPs have high passive tumor accumulation and that they can efficient co-deliver the two drugs to the tumor, reaching an 11-fold higher DOXO/DCTXL ratio in tumor as compared to free drugs. STATEMENT OF SIGNIFICANCE: Exploiting the synergistic action of multiple chemotherapeutics is a promising strategy to improve the outcome of cancer patients, as different agents can simultaneously engage different features of tumor cells and/or their microenvironment. Unfortunately, the choice is limited to drugs with similar pharmacokinetics that can concomitantly accumulate in tumors. To expand the spectrum of agents that can be delivered in combination, we propose a multi-compartmental core-shell nanoparticles approach, in which the core is made of biomaterials with high affinity for drugs of different physical properties. We successfully co-encapsulated Doxorubicin Hydrochloride, Docetaxel, and contrast agents and achieved a significantly higher concomitant accumulation in tumor versus free drugs, demonstrating that nanoparticles can improve synergistic cancer chemotherapy.

Honey possesses anti-bacterial, anti-inflammatory and other properties that are useful for wound healing and tissue regeneration. Furthermore, honey has been used for millennia in folk medicine. The misuse of antibiotics has again boosted the use of honey in regenerative medicine. The multifaceted properties of honey could possibly be exploited for scaffold applications in tissue healing.

The neurotransmitter glutamate increases cerebral blood flow by activating postsynaptic neurons and presynaptic glial cells within the neurovascular unit. Glutamate does so by causing an increase in intracellular Ca

Honey has successfully been used in the treatment of a broad spectrum of injuries including burns and non-healing wounds. It acts as an antibacterial and anti-biofilm agent with anti/pro-inflammatory properties. However, besides these traditional properties, recent evidence suggests that honey is also an immunomodulator in wound healing and contains several bee and plant-derived components that may speed up wound healing and tissue regeneration process. Identifying their exact mechanism of action allows better understanding of honey healing properties and promotes its wider translation into clinical practice.:This review will discuss the physiological basis for the use of honey in wound management, its current clinical uses, as well as the potential role of honey bioactive compounds in dermal regenerative medicine and tissue re-modeling.