6, 19. doi: 10.1166/jbn.2011.1312, Gangemi, S., Minciullo, P. L., Miroddi, M., Chinou, I., Calapai, G., and Schmidt, R. J. Histidine-tryptophan-ketoglutarate solution helps to preserve endothelial integrity of saphenous vein: an immunohistochemical and ultrastructural analysis. In particular, human dental pulp stem cells, represent a relevant cellular model constituted of a cell population homologous to the primary tissue with a large capacity of proliferation, the ability to maintain their cellular phenotype for a long time period, until 15 passage (Diomede et al., 2017a), and the possibility to undergo toward mesenchymal phenotype (Pizzicannella et al., 2018a). Oral Biol. The effects on the viability of hDPSCs treated with CurLIP, HEMA 3 and HEMA 5 mmol L1 were evaluated by means of the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide (MTT) method. RT-PCR provided an evidence of no increase of the above-mentioned genes. Sci. The polymerization is, often, incomplete, and induces the release of monomers into oral cavity, in dentin micro-channels and finally the bloodstream, inducing an inflammatory process. Curcumin modulates cytokines, growth factors, transcription factors, inflammatory molecules and cell signaling pathways. doi: 10.1177/039463201202500310, Trubiani, O., Caputi, S., Di Iorio, D., DAmario, M., Paludi, M., Giancola, R., et al. To study molecules involved in the intracellular signaling, hDPSCs treated with CurLIP, HEMA 3 and HEMA 5 mmol L1 were processed as previously reported by Giacoppo et al. (2008). doi: 10.1016/j.cyto.2017.04.022, Basnet, P., Hussain, H., Tho, I., and Skalko-Basnet, N. (2012). Photomed. doi: 10.1002/ptr.5640, Viale, M., Fontana, A., Maric, I., Monticone, M., Angelini, G., and Gasbarri, C. (2016). 51, 12531260. IFN synchronizes a diverse array of cellular programs through transcriptional regulation of immunologically relevant genes (Schroder et al., 2004). Design, preparation, and evaluation of liposomal gel formulations for treatment of acne: in vitro and in vivo studies. doi: 10.1016/j.joen.2013.06.004, Xiao, C. J., Yu, X. J., Xie, J. L., Liu, S., and Li, S. (2018). Front. (2015). Mutat. Biol. Int. p < 0.01, hDPSCs co-treated with curcumin and HEMA vs. HEMA treated hDPSCs. (2018). 28, 327339. 234, 22412251. doi: 10.1016/j.joen.2017.07.012, Diomede, F., Caputi, S., Merciaro, I., Frisone, S., DArcangelo, C., Piattelli, A., et al. The entrapment measurements confirmed an entrapment of 95.2% of guest into the bilayer (Figure 1B). (2018a). (2012). Dental Mater. Microbiol. CurLIP therapy in hDPSCs provokes an increase in cell proliferation and the block of inflammatory cytokines secretion through the inhibitory regulation of NFkB/ERK and pERK signaling cascade. Department of Precision Medicine, University of Campania Luigi Vanvitelli, Italy, Faculty of Medicine and Surgery, Marche Polytechnic University, Italy. Supernatants were read at 650 nm wavelength using a microplate reader (Synergy HT, BioTek Instruments, Winooski, VT, United States; Cavalcanti et al., 2015). MicroRNA 210 mediates VEGF upregulation in human periodontal ligament stem cells cultured on 3D hydroxyapatite ceramic scaffold. In vivo, the increased secretion of IL6 promotes monocyte differentiation into mobile and active macrophages able to secrete MCP1 and Metalloproteinases (MMPs), facilitating the invasion from the adventitia into the media (Kagan and Medzhitov, 2006) and it is also involved in root repair (Oliveira et al., 2018). J. Histochem. Endothelial committed oral stem cells as modelling in the relationship between periodontal and cardiovascular disease. doi: 10.1080/10717544.2018.1526227, Cheng, C., Peng, S. F., Li, Z. L., Zou, L. Q., Liu, W., and Liu, C. M. (2017). HEMA 3 and 5 mmol L1 stimulated an upregulation of IL6, IL8, MCP1, and IFN inflammatory molecules secretion, while CurLIP treatment significantly inhibited the expression of the above mentioned inflammatory cytokines. HEMA and TEGDMA impede odontogenic differentiation and the mineralization of apical papilla stem cells as well as the inflammatory cytokines secretion (Trubiani et al., 2010; Bakopoulou et al., 2012). Figure 5. Representative confocal images of NFkB (green), phalloidin 568 (green) and TOPRO (blue) fluorescent staining showed cells morphology in different culture conditions. Res. Diomede, F., Merciaro, I., Martinotti, S., Cavalcanti, M. F. X. Oral cavity is an easily accessible source of mesenchymal stem cells, as periodontal ligament, gingival and dental pulp (Stellavato et al., 2017). Stemness maintenance properties in human oral stem cells after long-term passage. Molecular mechanisms of hypolipidemic effects of curcumin. (2017). Int. 10, 8189. The pharmacokinetics of curcumin-loaded liposomal gel showed that nano-sized liposomes are able to penetrate within 1 h in both stratum of corneum and skin (Madan et al., 2018). Biochem. Bioavailab. 45, 1925. Axin2-expressing cells differentiate into reparative odontoblasts via autocrine Wnt/beta-catenin signaling in response to tooth damage. Nanocurcumin is superior to native curcumin in preventing degenerative changes in experimental cerebral malaria. J. CurLIP treatment restored the expression of NFkB at cytoplasmic level (Figures 5D,F). 33:50. doi: 10.1007/s11274-016-2195-0, Schroder, K., Hertzog, P. J., Ravasi, T., and Hume, D. A. Appl. 61, 122127. (2016). Table 1. (A) Untreated cells; (B) CurLIP treated cells; (C) HEMA 3 mmol L1 treated cells; (D) HEMA 3 mmol L1 with CurLIP treated cells; (E) HEMA 5 mmol L1 treated cells; (F) HEMA 5 mmol L1 with CurLIP treated cells. Resin monomers are released during the incomplete polymerization of composite resins and can penetrate into the pulp tissue exerting cytotoxic effects and inducing the release of proinflammatory cytokines. B. IL6 is a pleiotropic cytokine playing an active role in immune responses and in the development of the acute phase response (Heinrich et al., 1990). Progress in dentistry is associated with the innovation of dental materials and the use of new molecules for regenerative therapies, inter alia the design of new materials for restorative dentistry or for endodontic therapy. The analysis of released cytokines in the culture medium of the hDPSCs treated with CurLIP, HEMA 3 and HEMA 5 mmol L1 was performed by RayBiotech. 45, 395404. doi: 10.1016/j.bbrc.2008.01.077, Trubiani, O., Ballerini, P., Murmura, G., Pizzicannella, J., Giuliani, P., Buccella, S., et al. 7, 471475. Mag: 40. (2007). Then, findings indicate that CurLIP treatment is able to reduce inflammatory process via NFkB/ERK/pERK signaling, but it is not involved in the odontoblastic phenotype differentiation. hDPSCs spontaneously migrated from tissue and were cultured using MSCGM-CD medium (mesenchymal stem cell growth medium chemically defined; Lonza, Basel, Switzerland) maintaining in an incubator at 37C in a humidified atmosphere of 5% CO2 in air. 19:E3916. J. J. Mol. The zeta potential decreased on passing from pure POPC liposomes to curcumin-enriched liposomes, probably due to a partial deprotonation of phenolic moieties (pKa = 8.11 0.46, Calculated using Advanced Chemistry Development (ACD/Labs) Software V11.02) at the used buffered pH and the consequent tendency of dissociated phenoxide groups to expose toward the aqueous bulk. Endodont. Safety and anti-inflammatory activity of curcumin: a component of tumeric (Curcuma longa). 
J. Biol. (A) Untreated cells; (B) CurLIP treated cells; (C) HEMA 3 mmol L1 treated cells; (D) HEMA 3 mmol L1 with CurLIP treated cells; (E) HEMA 5 mmol L1 treated cells; (F) HEMA 5 mmol L1 with CurLIP treated cells. 9:419. doi: 10.3389/Fgene.2018.00419. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. Tuberous sclerosis complex 2 loss-of-function mutation regulates reactive oxygen species production through Rac1 activation. Following these evidences, we have analyzed, at molecular level, the involvement of CurLIP in odontoblast differentiation. Cell proliferation in combination with changes of the morphological features, proinflammatory cytokines secretion as Interleukin (IL) 6, IL8, Monocyte Chemoattractant Protein-1 (MCP1) and Interferon-gamma (IFN) were assayed along with the nuclear factor (NF)-kB, an inducible transcription factor involved in the activation of several cell processes associated to extracellular signal-regulated kinases (ERK) and posphorylated (p-) ERK pathway. RSC Adv. Physiol. Liposomes were prepared according to the above-mentioned protocol (Viale et al., 2016). Morphological analysis and interleukin release in human gingival fibroblasts seeded on different denture base acrylic resins. doi: 10.1016/j.cell.2006.03.047, Libro, R., Scionti, D., Diomede, F., Marchisio, M., Grassi, G., Pollastro, F., et al. Eur. Each data point represents the mean of three independent experiments. Studies carried out in other laboratories have identified a number of different molecules involved in inflammation that are inhibited by curcumin such as phospholipase, lipoxygenase, cyclooxygenase 2, leukotrienes, thromboxane, prostaglandins, nitric oxide, collagenase, elastase, hyaluronidase, monocyte chemoattractant protein-1 (MCP1), interferon-inducible protein, tumor necrosis factor (TNF), and interleukin (IL) 12 (Chainani-Wu, 2003). J. Tissue Eng. Pizzicannella, J., Rabozzi, R., Trubiani, O., and Di Giammarco, G. (2011). J. Mol. Res. Cell. Potential effects of curcumin on peroxisome proliferator-activated receptor-gamma in vitro and in vivo. Rep. 7:10062. doi: 10.1038/S41598-017-10672-9, Deshpande, A. S., Fang, P. A., Zhang, X. Y., Jayaraman, T., Sfeir, C., and Beniash, E. (2011). doi: 10.1080/03639045.2018.1546310, Mammana, S., Gugliandolo, A., Cavalli, E., Diomede, F., Iori, R., Zappacosta, R., et al. The variations in NFkB, ERK and pERK protein expression were further evaluated by the western blot analysis after treatment. (2019). 7, 188192. doi: 10.1002/jbm.a.36213. doi: 10.1159/000485414, Suzuki, S., Haruyama, N., Nishimura, F., and Kulkarni, A. IL6 and IL8 cytokines secretion is related to increased activation of the transcription factor NFkB. However, uncontrolled inflammation is the cause of a wide range of pathological conditions, including cardiovascular diseases, psoriasis, multiple sclerosis, rheumatoid arthritis and inflammatory bowel disease (Soltani et al., 2019). DSPP is a positive regulators of hard tissue mineralization (Suzuki et al., 2012), acting as nucleators of apatite crystal formation in the presence of collagen. RUNX2 and COL1A1 possess a decisive role during mineralization process in the early step of mineralized tissue formation (Diomede et al., 2016b). 
Pharmacol. Liposomes were sterilized under UV lamp for 2 h. Then an appropriate amount of curcumin in DMSO was added to the resulting liposomal suspension in order to obtain a POPC to curcumin molar ratio of 25:1. B., Caputi, S., Mazzon, E., et al. MDA and VG performed RT-PCR analysis and revised the manuscript. Part A 106, 126137. Micrographs were obtained using excitation lines at 488 nm for argon laser beam and at 543 and 665 nm for a helium-neon source. Figure 3. An appropriate aliquot of POPC (1-palmitoyl-2-oleoyl-phosphatidylcholine, Avanti Polar Lipids, Alabama, United States), dissolved in chloroform, was put in a round-bottomed flask and dried in a rotary evaporator under reduced pressure at 40C to form a thin lipid film on the inside wall of the flask. Reduction of cell proliferation and alteration of cell morphology have been directly associated with the HEMA treatment (Trubiani et al., 2010). 44, 10781092. J. Bioequivalence. doi: 10.1177/1721727x1201000109, Trubiani, O., Cataldi, A., De Angelis, F., DArcangelo, C., and Caputi, S. (2012b). Int. Clin. Interferon-gamma: an overview of signals, mechanisms and functions. J. Endodont. Head Face Med. Human DPSCs fabricate vascularized woven bone tissue: a new tool in bone tissue engineering. by curcumin. There is an increasing interest in the tissues regeneration for the use of autologous stem cells isolated from various adult tissues due to the fewer ethical concerns as compared to embryonic stem cells (Spina et al., 2016). The phospholipid film was kept at 4C overnight before rehydration with PBS buffer (pH 7.4) and sonication for 30 min (Viale et al., 2016; Mammana et al., 2019). (2019). Curcumin attenuates diet-induced hepatic steatosis by activating AMP-Activated protein kinase. RNA quantity and quality were assessed by Qubit 2.0 (ThermoFisher Scientific, Whaltam, MA, United States; Diomede et al., 2018b). (A) Untreated cells; (B) CurLIP treated cells; (C) HEMA 3 mmol L1 treated cells; (D) HEMA 3 mmol L1 with CurLIP treated cells; (E) HEMA 5 mmol L1 treated cells; (F) HEMA 5 mmol L1 with CurLIP treated cells. J. Biomed. Agents 30, 10091018. Front. Physiol. Commun. qRT-PCR analysis showed no significant differences among the analyzed conditions for all tested genes. (2012). Cytokine 96, 261272. All experiments were performed in triplicate. Cells were treated with HEMA 3 and HEMA 5 mmol L1 for 48h and pretreated with CurLIP for 24h. The natural nanocarrier CurLIP influences numerous biochemical and molecular cascades causing anti-inflammatory properties in response to HEMA treatment in human dental pulp stem cells, representing an innovative endodontic formulation able to improve the quality of dental care with a major human community impact. Curcumin is pharmacologically safe. 97, 1017710187. The editor and reviewer's affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. doi: 10.1089/pho.2014.3864, Chainani-Wu, N. (2003). doi: 10.1111/bcpt.12076, Vaughn, A. R., Branum, A., and Sivamani, R. K. (2016). Physiol. (2012a). 9, 161168. J. Cancer 31, 243305. 5-Aza exposure improves reprogramming process through embryoid body formation in human gingival stem cells. 14, 12. doi: 10.1186/s13005-018-0169-1, Zingg, J. M., Hasan, S. T., and Meydani, M. (2013). Int. (2016). Mag: 40. 6, 112117. (2018). doi: 10.1002/jcp.26515, Pizzicannella, J., Gugliandolo, A., Orsini, T., Fontana, A., Ventrella, A., Mazzon, E., et al. Evaluation of the proliferative effects induced by low-level laser therapy in bone marrow stem cell culture. 5-Adenosine monophosphate-activated protein kinase: a potential target for disease prevention by curcumin. doi: 10.1016/j.mrgentox.2011.04.011, PubMed Abstract | CrossRef Full Text | Google Scholar, Babb, R., Chandrasekaran, D., Neves, V. C. M., and Sharpe, P. T. (2017). miR-2861 is involved in osteogenic commitment of human periodontal ligament stem cells grown onto 3d scaffold. doi: 10.1111/j.1751-1097.1994.tb05037.x, Dende, C., Meena, J., Nagarajan, P., Nagaraj, V. A., Panda, A. K., and Padmanaban, G. (2017). BS, JP, and FD designed the study, performed the experiments, analyzed the data, and wrote the manuscript. 33, 377390. doi: 10.1111/j.1365-2591.2010.01720.x, Um, M. Y., Hwang, K. H., Ahn, J., and Ha, T. Y. World J. Microbiol. Toll-Like Receptor 4 Expression, Interleukin-6,-8 and Ccl-20 Release, and Nf-Kb translocation in human periodontal ligament mesenchymal stem cells stimulated with Lps-P-Gingivalis. (A) The western blotting analysis with anti-NFkB, ERK and pERK in treated and untreated cells. BioFactors 39, 101121. Resin-based methacrylates, common materials widely used in restorative dentistry, are viscous substances that are converted into solid material trough polymerization processes (Trubiani et al., 2012c). This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). Sci. -actin as internal control is shown. This data offers the opportunity to speculate that CurLIP is able to provide a niche environment for cells replacement, as demonstrated through the increase of cell proliferation. Previously we have demonstrated that LPS-Gingivalis (G) inflammatory stimulus provokes, in endothelial cells, the release of IL6, IL8, and MCP1 molecules and the activation of signaling network Toll Like Receptor 4 (TLR4)/NFkB/ERK/pERK (Diomede et al., 2017b; Pizzicannella et al., 2018b). 47, 864872. On the other hand, cells subjected to the combined treatment with CurLIP/HEMA 3 or 5 mmol L1 showed a proliferation rate similar to the control cells. Rep. 7:3102. doi: 10.1038/S41598-017-03145-6, Bakopoulou, A., Leyhausen, G., Volk, J., Koidis, P., and Geurtsen, W. (2012). 57, 11651175. Mag: 40. Homeostat. This arrangement, in line with the recently proposed pH-driven, organic solvent-free, liposomal encapsulation of curcumin in liposomes (Cheng et al., 2017), was particularly pronounced in the present experimental conditions as compared to previously investigated curcumin loaded liposomes because we chose to add the curcumin on preformed liposomes.