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Graphene quantum dots (GQDs) are unique derivatives of graphene that show promise in multiple biomedical applications as biosensors, bioimaging agents, and drug/gene delivery vehicles. Their ease in functionalization, biocompatibility, and intrinsic fluorescence enable those modalities. However, GQDs lack deep tissue magnetic resonance imaging (MRI) capabilities desirable for diagnostics. Considering that the drawbacks of MRI contrast agent toxicity are still poorly addressed, we develop novel Mn2+ or Gd3+ doped nitrogen-containing graphene quantum dots (NGQDs) to equip the GQDs with MRI capabilities and at the same time render contrast agents biocompatible. Water-soluble biocompatible Mn-NGQDs and Gd-NGQDs synthesized via single-step microwave-assisted scalable hydrothermal reaction enable dual MRI and fluorescence modalities. These quasi-spherical 3.9-6.6 nm average-sized structures possess highly crystalline graphitic lattice structure with 0.24 and 0.53 atomic % for Mn2+ and Gd3+ doping. This structure ensures high in vitro biocompatibility of up to 1.3 mg ml-1 and 1.5 mg ml-1 for Mn-NGQDs and Gd-NGQDs, respectively, and effective internalization in HEK-293 cells traced by intrinsic NGQD fluorescence. As MRI contrast agents with considerably low Gd and Mn content, Mn-NGQDs exhibit substantial transverse/longitudinal relaxivity (r 2/r 1) ratios of 11.190, showing potential as dual-mode longitudinal or transverse relaxation time (T 1 or T 2) contrast agents, while Gd-NGQDs possess r 2/r 1 of 1.148 with high r 1 of 9.546 mM-1 s-1 compared to commercial contrast agents, suggesting their potential as T1 contrast agents. Compared to other nanoplatforms, these novel Mn2+ and Gd3+ doped NGQDs not only provide scalable biocompatible alternatives as T1/T2 and T1 contrast agents but also enable in vitro intrinsic fluorescence imaging.
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This work develops a new multifunctional biocompatible anticancer nanoformulation to provide targeted image-guided cancer-selective therapeutics. It consists of three active covalently bound components: (1) biocompatible nitrogen-doped graphene quantum dots (GQDs) as a multifunctional delivery and imaging platform, (2) hyaluronic acid (HA) unit targeted to the CD44 receptors on a variety of cancer cells, and (3) oxidative stress-based cancer-selective ferrocene (Fc) therapeutic. The biocompatible GQD platform synthesized from glucosamine exhibits high-yield intrinsic fluorescence. It is utilized for tracking Fc-GQD-HA formulation in vitro indicating internalization enhancement in HeLa cells targeted by the HA over non-cancer HEK-293 cells not overexpressing CD44 receptor. Fc-GQD-HA, non-toxic at 1 mg/mL to HEK-293 cells, induces cytotoxic response in HeLa enhanced over time, while therapeutic ROS generation by Fc-GQD-HA is ~3 times greater than that of Fc alone. This outlines the targeted delivery, imaging, and cancer-specific treatment capabilities of the new Fc-GQD-HA formulation enabling desired cancer-focused nanotherapeutic approach.
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Sistemas de Liberação de Medicamentos , Grafite/farmacologia , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Antineoplásicos/química , Antineoplásicos/farmacologia , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Doxorrubicina/química , Doxorrubicina/farmacologia , Grafite/química , Células HEK293 , Células HeLa , Humanos , Receptores de Hialuronatos/antagonistas & inibidores , Receptores de Hialuronatos/genética , Ácido Hialurônico/química , Ácido Hialurônico/farmacologia , Neoplasias/genética , Neoplasias/patologia , Imagem Óptica , Oxirredução/efeitos dos fármacos , Pontos Quânticos/químicaRESUMO
With the advent of graphene, there has been an interest in utilizing this material and its derivative, graphene oxide (GO) for novel applications in nanodevices such as bio and gas sensors, solid-state supercapacitors and solar cells. Although GO exhibits lower conductivity and structural stability, it possesses an energy band gap that enables fluorescence emission in the visible/near infrared leading to a plethora of optoelectronic applications. In order to allow fine-tuning of its optical properties in the device geometry, new physical techniques are required that, unlike existing chemical approaches, yield substantial alteration of GO structure. Such a desired new technique is one that is electronically controlled and leads to reversible changes in GO optoelectronic properties. In this work, we for the first time investigate the methods to controllably alter the optical response of GO with the electric field and provide theoretical modeling of the electric field-induced changes. Field-dependent GO emission is studied in bulk GO/polyvinylpyrrolidone films with up to 6% reversible decrease under 1.6 V µm-1 electric fields. On an individual flake level, a more substantial over 50% quenching is achieved for select GO flakes in a polymeric matrix between interdigitated microelectrodes subject to two orders of magnitude higher fields. This effect is modeled on a single exciton level by utilizing Wentzel, Kremer, and Brillouin approximation for electron escape from the exciton potential well. In an aqueous suspension at low fields, GO flakes exhibit electrophoretic migration, indicating a degree of charge separation and a possibility of manipulating GO materials on a single-flake level to assemble electric field-controlled microelectronics. As a result of this work, we suggest the potential of varying the optical and electronic properties of GO via the electric field for the advancement and control over its optoelectronic device applications.
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To estimate daily intake of total phenolics and flavonoids from green tea and the contribution of green tea to the antioxidant intake from the Korean diet, 24 commercial brands of green tea were selected and analyzed. Data from the Korea National Health and Nutrition Examination Survey (KNHANES) from 2008 and 2011 indicate that the green tea consumption in these 2 years was 2.8 g/tea drinker/day and 2.9 g/tea drinker/day, respectively. Based on data derived from direct measurements of green tea phenolics and the dataset of the 2008 KNHANES, we estimated the daily per tea drinker phenolics intake to be 172 mg gallic acid equivalents (GAE), the total flavonoids to be 43 mg catechin equivalents (CE) and the total antioxidants to be 267 mg vitamin C equivalents (VCE; 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay) and 401 mg VCE (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assay). In 2011, we estimated the daily per tea drinker total phenolics intake to be 246 mg GAE, the total flavonoids to be 60 mg CE and the antioxidants to be 448 mg VCE (DPPH assay) and 630 mg VCE (ABTS assay). The daily intake of total phenolics, total flavonoids and antioxidants from green tea consumption increased from 2008 to 2011.
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Antioxidantes/administração & dosagem , Fenóis/administração & dosagem , Chá/química , Antioxidantes/química , Benzotiazóis , Compostos de Bifenilo , Dieta , Comportamento Alimentar , Flavonoides/administração & dosagem , Flavonoides/química , Humanos , Inquéritos Nutricionais , Fenóis/química , Picratos , República da Coreia , Ácidos SulfônicosRESUMO
Oxidative stress is proven to be a leading factor in a multitude of adverse conditions, from Alzheimer's disease to cancer. Thus, developing effective radical scavenging agents to eliminate reactive oxygen species (ROS) driving many oxidative processes has become critical. In addition to conventional antioxidants, nanoscale structures and metal-organic complexes have recently shown promising potential for radical scavenging. To design an optimal nanoscale ROS scavenging agent, we have synthesized ten types of biocompatible graphene quantum dots (GQDs) augmented with various metal dopants. The radical scavenging abilities of these novel metal-doped GQD structures were, for the first time, assessed via the DPPH, KMnO4, and RHB (Rhodamine B protectant) assays. While all metal-doped GQDs consistently demonstrate antioxidant properties higher than the undoped cores, aluminum-doped GQDs exhibit 60-95% radical scavenging ability of ascorbic acid positive control. Tm-doped GQDs match the radical scavenging properties of ascorbic acid in the KMnO4 assay. All doped GQD structures possess fluorescence imaging capabilities that enable their tracking in vitro, ensuring their successful cellular internalization. Given such multifunctionality, biocompatible doped GQD antioxidants can become prospective candidates for multimodal therapeutics, including the reduction of ROS with concomitant imaging and therapeutic delivery to cancer tumors.
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While small interfering RNA (siRNA) technology has become a powerful tool that can enable cancer-specific gene therapy, its translation to the clinic is still hampered by the inability of the genes alone to cell transfection, poor siRNA stability in blood, and the lack of delivery tracking capabilities. Recently, graphene quantum dots (GQDs) have emerged as a novel platform allowing targeted drug delivery and fluorescence image tracking in visible and near-infrared regions. These capabilities can aid in overcoming primary obstacles to siRNA therapeutics. Here, for the first time, we utilize biocompatible nitrogen- and neodymium-doped graphene quantum dots (NGQDs and Nd-NGQDs, respectively) for the delivery of Kirsten rat sarcoma virus (KRAS) and epidermal growth factor receptor (EGFR) siRNA effective against a variety of cancer types. GQDs loaded with siRNA noncovalently facilitate successful siRNA transfection into HeLa cells, confirmed by confocal fluorescence microscopy at biocompatible GQD concentrations of 375 µg/mL. While the GQD platform provides visible fluorescence tracking, Nd doping enables deeper-tissue near-infrared fluorescence imaging suitable for both in vitro and in vivo applications. The therapeutic efficacy of the GQD/siRNA complex is verified by successful protein knockdown in HeLa cells at nanomolar siEGFR and siKRAS concentrations. A range of GQD/siRNA loading ratios and payloads are tested to ultimately provide substantial inhibition of protein expression down to 31-45%, comparable with conventional Lipofectamine-mediated delivery. This demonstrates the promising potential of GQDs for the nontoxic delivery of siRNA and genes in general, complemented by multiwavelength image tracking.
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Grafite , Neoplasias , Pontos Quânticos , Humanos , Células HeLa , Neodímio , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/uso terapêutico , NitrogênioRESUMO
Sonography offers many advantages over standard methods of diagnostic imaging due to its non-invasiveness, substantial tissue penetration depth, and low cost. The benefits of ultrasound imaging call for the development of ultrasound-trackable drug delivery vehicles that can address a variety of therapeutic targets. One disadvantage of the technique is the lack of high-precision imaging, which can be circumvented by complementing ultrasound contrast agents with visible and, especially, near-infrared (NIR) fluorophores. In this work, we, for the first time, develop a variety of lightly metal-doped (iron oxide, silver, thulium, neodymium, cerium oxide, cerium chloride, and molybdenum disulfide) nitrogen-containing graphene quantum dots (NGQDs) that demonstrate high-contrast properties in the ultrasound brightness mode and exhibit visible and/or near-infrared fluorescence imaging capabilities. NGQDs synthesized from glucosamine precursors with only a few percent metal doping do not introduce additional toxicity in vitro, yielding over 80% cell viability up to 2 mg/mL doses. Their small (<50 nm) sizes warrant effective cell internalization, while oxygen-containing surface functional groups decorating their surfaces render NGQDs water soluble and allow for the attachment of therapeutics and targeting agents. Utilizing visible and/or NIR fluorescence, we demonstrate that metal-doped NGQDs experience maximum accumulation within the HEK-293 cells 6-12 h after treatment. The successful 10-fold ultrasound signal enhancement is observed at 0.5-1.6 mg/mL for most metal-doped NGQDs in the vascular phantom, agarose gel, and animal tissue. A combination of non-invasive ultrasound imaging with capabilities of high-precision fluorescence tracking makes these metal-doped NGQDs a viable agent for a variety of theragnostic applications.
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Grafite , Pontos Quânticos , Animais , Humanos , Células HEK293 , Nitrogênio , Imagem Óptica , UltrassonografiaRESUMO
Phenolic compounds in common buckwheat sprouts (CBSs) have gained research interest because of their multiple health benefits. Phenolic acids, flavanones, flavonols, flavan-3-ols, and anthocyanins are important bioactive components of CBS that exhibit biological activities, including anti-inflammatory, antioxidant, anti-proliferative, and immunomodulatory effects. The isolation and quantitative and qualitative analyses of these phenolic compounds require effective and appropriate extraction and analytical methods. The most recent analytical method developed for determining the phenolic profile is HPLC coupled with a UV-visible detector and/or MS. This review highlights the extraction, purification, analysis, and bioactive properties of phenolic compounds from CBS described in the literature.
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Non-invasive temperature sensing is necessary to analyze biological processes occurring in the human body, including cellular enzyme activity, protein expression, and ion regulation. To probe temperature-sensitive processes at the nanoscale, novel luminescence nanothermometers are developed based on graphene quantum dots (GQDs) synthesized via top-down (RGQDs) and bottom-up (N-GQDs) approaches from reduced graphene oxide and glucosamine precursors, respectively. Because of their small 3-6 nm size, non-invasive optical sensitivity to temperature change, and high biocompatibility, GQDs enable biologically safe sub-cellular resolution sensing. Both GQD types exhibit temperature-sensitive yet photostable fluorescence in the visible and near-infrared for RGQDs, utilized as a sensing mechanism in this work. Distinctive linear and reversible fluorescence quenching by up to 19.3% is observed for the visible and near-infrared GQD emission in aqueous suspension from 25 °C to 49 °C. A more pronounced trend is observed with GQD nanothermometers internalized into the cytoplasm of HeLa cells as they are tested in vitro from 25 °C to 45 °C with over 40% quenching response. Our findings suggest that the temperature-dependent fluorescence quenching of bottom-up and top-down-synthesized GQDs studied in this work can serve as non-invasive reversible/photostable deterministic mechanisms for temperature sensing in microscopic sub-cellular biological environments.
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With 18 million new cases diagnosed each year worldwide, cancer strongly impacts both science and society. Current models of cancer cell growth and therapeutic efficacy in vitro are time-dependent and often do not consider the Emax value (the maximum reduction in the growth rate), leading to inconsistencies in the obtained IC50 (concentration of the drug at half maximum effect). In this work, we introduce a new dual experimental/modeling approach to model HeLa and MCF-7 cancer cell growth and assess the efficacy of doxorubicin chemotherapeutics, whether alone or delivered by novel nitrogen-doped graphene quantum dots (N-GQDs). These biocompatible/biodegradable nanoparticles were used for the first time in this work for the delivery and fluorescence tracking of doxorubicin, ultimately decreasing its IC50 by over 1.5 and allowing for the use of up to 10 times lower doses of the drug to achieve the same therapeutic effect. Based on the experimental in vitro studies with nanomaterial-delivered chemotherapy, we also developed a method of cancer cell growth modeling that (1) includes an Emax value, which is often not characterized, and (2), most importantly, is measurement time-independent. This will allow for the more consistent assessment of the efficiency of anti-cancer drugs and nanomaterial-delivered formulations, as well as efficacy improvements of nanomaterial delivery.
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Quantum dots (QDs) are semiconductor nanoparticles ranging in size from 2 to 10 nm. QDs are increasingly being developed for biomedical imaging, targeted drug delivery, and green energy technology. Here we describe the novel utilization of biocompatible CdSe-ZnS core-shell semiconductor nanoparticles for assessment of reactive oxygen species (ROS) in the context of chemotherapy and radiotherapy, both of which are important modalities in the treatment of cancer.
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Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/metabolismo , Pontos Quânticos/química , Espécies Reativas de Oxigênio/análise , Compostos de Cádmio/química , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Tratamento Farmacológico/métodos , Humanos , Radioterapia/efeitos adversos , Espécies Reativas de Oxigênio/química , Compostos de Selênio/química , Sulfetos/química , Compostos de Zinco/químicaRESUMO
Near-infrared (NIR) fluorescence provides a new avenue for biomedical fluorescence imaging that allows for the tracking of fluorophore through several centimeters of biological tissue. However, such fluorophores are rare and, due to accumulation-derived toxicity, are often restricted from clinical applications. Deep tissue imaging not only provided by near-infrared fluorophores but also conventionally carried out by magnetic resonance imaging (MRI) or computed tomography (CT) is also hampered by the toxicity of the contrast agents. This work offers a biocompatible imaging solution: cerium oxide (CeO2) nanocubes doped with ytterbium or neodymium, and co-doped with gadolinium, showing simultaneous potential for near-infrared (NIR) fluorescence and magnetic resonance imaging (MRI) applications. A synthetic process described in this work allows for the stable incorporation of ytterbium or neodymium, both possessing emissive transitions in the NIR. As a biocompatible nanomaterial, the CeO2 nanocubes act as an ideal host material for doping, minimizing lanthanide fluorescence self-quenching as well as any potential toxicity associated with the dopants. The uptake of nanocubes by HeLa cells maximized at 12 h was monitored by hyperspectral imaging of the ytterbium or neodymium NIR emission, indicating the capacity of the lanthanide-doped nanocubes for in vitro and a potential for in vivo fluorescence imaging. The co-doped nanocubes demonstrate no significant loss of NIR emission intensity upon co-doping with 2 atomic % gadolinium and exhibit magnetic susceptibilities in the range of known negative contrast agents. However, a small increase to 6 atomic % gadolinium significantly affects the magnetic susceptibility ratio (r2/r1), shifting closer to the positive contrast range and suggesting the potential use of the CeO2 nanocube matrix doped with selected rare-earth ions as a tunable MRI contrast agent with NIR imaging capabilities.
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Cério , Metais Terras Raras , Células HeLa , Humanos , Imageamento por Ressonância MagnéticaRESUMO
This study evaluated the daily per drinker intakes of total phenolics, total flavonoids, and antioxidants from coffee in the Korean diet. Four types of coffee (instant coffee, decaffeinated instant coffee, roasted coffee, and coffee mix) were selected and analyzed. Based on the Korea National Health and Nutrition Examination Survey in 2012, the daily intakes per coffee drinker were estimated to be 348.6 mg gallic acid equivalents for phenolics, 222.5 mg catechin equivalents for flavonoids, and 370.8 mg vitamin C equivalents (DPPH assay) or 546.7 mg vitamin C equivalents (ABTS assay) for antioxidants. Men had higher intakes of instant coffee and coffee mix, while women had a higher intake of roasted coffee. Regarding age categories, over 58% of the coffee consumers were 30-59 years old. This study revealed that, in Korea, approximately half of the people drank about 1.4 cups of roasted coffee or 2.0 cups of instant coffee every day.
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Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated H2O2-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of H2O2-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against H2O2-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against H2O2-induced neurotoxicity through their antioxidant properties.
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Sobrevivência Celular/efeitos dos fármacos , Flavonoides/farmacologia , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Rhus/química , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Benzofuranos/farmacologia , Caspase 3/metabolismo , Chalconas/farmacologia , Ditiocarb/farmacologia , Flavonoides/química , Flavonoides/isolamento & purificação , Neurônios/efeitos dos fármacos , Células PC12 , Fenóis/farmacologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Ratos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Superóxido Dismutase/metabolismoRESUMO
Buckwheat sprouts have been widely consumed all around world due to their great abundance of bioactive compounds. In this study, the anti-inflammatory effects of flavonoid-rich common buckwheat sprout (CBS) and tartary buckwheat sprout (TBS) extracts were evaluated in lipopolysaccharide- (LPS-) stimulated RAW 264.7 murine macrophages and primary peritoneal macrophages from male BALB/c mice. Based on the reversed-phase HPLC analysis, the major flavonoids in CBS were determined to be C-glycosylflavones (orientin, isoorientin, vitexin, and isovitexin), quercetin-3-O-robinobioside, and rutin, whereas TBS contained only high amounts of rutin. The TBS extract exhibited higher inhibitory activity as assessed by the production of proinflammatory mediators such as nitric oxide and cytokines including tumor necrosis factor-α, interleukin- (IL-) 6, and IL-12 in LPS-stimulated RAW 264.7 macrophages than CBS extract. In addition, TBS extract suppressed nuclear factor-kappa B activation by preventing inhibitor kappa B-alpha degradation and mitogen-activated protein kinase phosphorylation in LPS-stimulated RAW 264.7 macrophages. Moreover, the TBS extract markedly reduced LPS-induced cytokine production in peritoneal macrophages. Taken together, these findings suggest that TBS extract can be a potential source of anti-inflammatory agents that may influence macrophage-mediated inflammatory disorders.
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Anti-Inflamatórios/farmacologia , Fagopyrum/química , Flavonoides/farmacologia , Macrófagos Peritoneais/metabolismo , Extratos Vegetais/farmacologia , Animais , Cromatografia Líquida de Alta Pressão , Ciclo-Oxigenase 2/metabolismo , Citocinas/biossíntese , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/enzimologia , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Células RAW 264.7 , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Oxidative stress damages dermal and epidermal cells and degrades extracellular matrix proteins, such as collagen, ultimately leading to skin aging. The present study evaluated the potential protective effect of the aqueous methanolic extract obtained from Lithospermum erythrorhizon (LE) against oxidative stress, induced by H2O2 and ultraviolet (UV) irradiation, on human keratinocyte (HaCaT) and human dermal fibroblast-neonatal (HDF-n) cells. Exposure of cells to H2O2 or UVB irradiation markedly increased oxidative stress and reduced cell viability. However, pretreatment of cells with the LE extract not only increased cell viability (up to 84.5%), but also significantly decreased oxidative stress. Further, the LE extract downregulated the expression of matrix metalloproteinase-1, an endopeptidase that degrades extracellular matrix collagen. In contrast, treatment with the LE extract did not affect the expression of procollagen type 1 in HDF-n cells exposed to UVA irradiation. Thirteen phenolic compounds, including derivatives of shikonin and caffeic acid, were identified by ultrahigh-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. These results suggest that LE-derived extracts may protect oxidative-stress-induced skin aging by inhibiting degradation of skin collagen, and that this protection may derive at least in part from the antioxidant phenolics present in these extracts. Further studies are warranted to determine the potential utility of LE-derived extracts in both therapeutic and cosmetic applications.