Content uploaded by Hisae Aoshima
Author content
All content in this area was uploaded by Hisae Aoshima on Nov 06, 2014
Content may be subject to copyright.
A preview of the PDF is not available
Defensive Effects of Fullerene-C60 Dissolved in Squalane Against the 2,4-Nonadienal-Induced Cell Injury in Human Skin Keratinocytes HaCaT and Wrinkle Formation in 3D-Human Skin Tissue Model
Abstract
We dissolved fullerene-C60 in squalane (LipoFullerene; LF-SQ, C60-eq.: 500 ppm) and examined its defensive effects against 2,4-nonadienal (NDA)-induced cell injury in HaCaT keratinocytes and wrinkle formation in three dimensional (3D)-human skin tissue model. NDA is an analog of 4-hydroxynonenal, one of major causes for human body odor indicative of aging and a lipophilic cell injury factor. Cell viability (% of the control) decreased to 31.6% on treatment with NDA (40 microM), but it increased to 66.0-97.5% when LF-SQ of 1-4% (C60-eq.: 5-20 ppm) was administered for 5 hr before NDA addition. The defensive effect by LF-SQ was superior to that of "squalane" alone at the same doses. NDA-induced DNA-fragmentation in HaCaT cells was suppressed by LF-SQ administered for 5 hr before NDA treatment, and LF-SQ protected HaCaT cells against apoptosis-like cell death. LF-SQ did not appreciably defend against hydrogen peroxide, though LF-SQ effectively defended against tert-butylhydroperoxide, a type of the intermediate hydrophilicity-lipophilicity degree out of other reactive oxygen species. The scanning electron microscopy demonstrated that NDA caused wrinkles and abnormal scales on keratinocytes of 3D-human skin tissue model, and structural homogeneity of the interstratum was broken, any of which were, however, markedly suppressed with LF-SQ. Squalane alone exhibited defensive effect against the skin tissue injury to some extent, but which was inferior to LF-SQ. LF-SQ might effectively capture and scavenge lipid radicals generated inside the cell membrane, because squalane acts as a lipophilic carrier of C60. C60 dissolved in squalane can be expected to serve as a cosmeceutical ingredient for anti-wrinkle formation.
... 44 Numerous studies have pointed out the protective effect of olive tree compounds on skin ageing, thanks to their role in the various mechanisms involved in the ageing process. [45][46][47] These compounds include squalene (Sq) from olives or their derivatives. In an in vitro test with HaCat keratinocytes, Kato et al. 45 observed a significant reduction in oxidative stress, an increase in cell viability, and a reduction in histological alterations in a 3D human skin model, which could translate into a protective effect against the appearance of signs of ageing. ...
... [45][46][47] These compounds include squalene (Sq) from olives or their derivatives. In an in vitro test with HaCat keratinocytes, Kato et al. 45 observed a significant reduction in oxidative stress, an increase in cell viability, and a reduction in histological alterations in a 3D human skin model, which could translate into a protective effect against the appearance of signs of ageing. The same authors studied the antioxidant effect of Sq compared to a solution of Sq and C-60 fullerene in a clinical trial, analysing wrinkle formation and skin hydration. ...
... This could be due to impaired nuclear factor-kappaB signaling and could reflect in the decreased degradation of extracellular matrix components. 50 However, the anti-ageing effect derived from the compounds present in the olive tree is not limited to its topical 45 ...
The olive tree and its derivatives are of great interest in the field of biomedicine due to their numerous health properties. The aim of the present study was to identify the effects of the use of olive products, extra virgin olive oil (EVOO) and products derived from its extraction, on the skin. Numerous studies have pointed out the protective effect of olive compounds on skin ageing, thanks to their role in the different mechanisms involved in the ageing process, such as reducing oxidative stress, increasing cell viability and decreasing histological alterations. With regard to their photoprotective effect, the olive tree and its fruit contain phenolic compounds which have a protective effect against radiation, such as low ultraviolet absorption and high antioxidant activity, acting as a protective factor against photocarcinogenesis. Similarly, the anti-tumour effects of olives have been studied at the level of the different compounds and extracts obtained from them, and their ability to selectively attack human melanoma cells has been observed. They have also shown antibacterial activity against microorganisms particularly implicated in skin infections, such as Escherichia coli, Candida albicans, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Pseudomonas aeruginosa and Proteus spp. Likewise, on healthy tissue, they have shown the ability to stimulate growth, migration and the expression of genes involved in cell differentiation, which favours the regeneration of skin wounds. According to the results included in this review, the olive tree and its derivatives could be useful in the treatment of many skin conditions.
... Since the action is frequently limited to the top layers of the skin when used for cosmetic purposes. [116][117] Retinol Amplify the manufacture of collagen, cell renewal, epidermal hyperplasia, and water content in the skin. ...
Cosmetics attract most age groups from teenagers to old age. Cosmetic goods now contain a variety of nanoparticle and nanomaterial types. Nano cosmeceuticals have changed the era of cosmetics as they have advanced delivery mechanisms with task specifications. They are used in nail, hair, lip, and skin care products by cosmetic giants including Estee Lauder, L’oreal, Nivea, Zelens, and Derma Swiss, etc., and have patented the use of dozens of "nanosome particles.” The global market for cosmetics using nanotechnology is worth millions of dollars and increasing at 7.14% annually. Liposomes, niosomes,nanostructured lipid carriers, solid lipid nanoparticles, gold nanoparticles, nanoemulsions, and nanosomes are novel nanocarriers that are now used in a variety of cosmeceuticals for drug delivery to achieve site specification, improved stability, biocompatibility, extended action, and increased drug-loading capacity. In this chapter use of various nanocarriers in cosmetic applications with their safety concerns will be discussed.
... Фуллерен в виде комплекса С 60 /ПВП или раствора в сквалане эффективно защищает клеточную мембрану от перекисного окисления, поэтому можно также ожидать, что фуллерен, как антиоксидант, будет удалять АФК, образующиеся на поверхности кожи при использовании TiO 2 -содержащих солнцезащитных кремов [54][55][56]. ...
The review focuses on the possibilities and prospects of the use of fullerenes and their derivatives in cosmetics, the only industrial area where fullerenes have found practical application today. Based on the literary data and the results of their own experiments, the authors substantiate the safety of using fullerene for living organisms, as well as the usefulness of introducing fullerene as antioxidant in cosmetic compositions. Other useful properties of fullerene used in cosmetics and dermatology are discussed.
... To sum up, 3D cell culture shows incomparable advantages in simulating tumor H. Z. Yang, J. Q. Jiao Superior defensive and anti-wrinkle actions of squalane loaded fullerene-C60 was reported [19] microenvironment, nanodrug delivery and drug screening. At present, a variety of bio-simulated 3D tumor models have been widely used, such as the 3D culture resistance model of breast cancer cells [20], the 3D culture model of OSCC-3 [21], the 3D skin model [22], and the 3D blood and brain barrier model in vitro [23]. ...
Nanotechnology is a relatively new field of engineering that has led to the development of nanomedicine. Many of the benefits and risks surrounding nanotechnology concern the skin, and research over the last few decades has led to a better understanding of the potential role for nanotechnology in dermatology. As this field is rapidly evolving, practitioners must stay up to date on current research. This chapter will discuss the current uses and future direction of nanomaterials in cosmetic, diagnostic, and therapeutic dermatology.
Целью исследования было определение влияния степени переработки пищевого растительного масла на его устойчивость к окислению, формирование вторичных продуктов окисления и технологических контаминантов - сложных эфиров монохлорпропандиолов и глицидола с жирными кислотами, свободные формы которых включены в группы «возможных канцерогенов человека» 2B и 2А соответственно. Проведена комплексная гигиеническая оценка влияния технологических процессов очистки пищевых растительных масел, в том числе проведения многократных дезодораций на качество, безопасность и сохранность конечного продукта. В том числе было проведено изучение изменений химического состава пищевых подсолнечных масел в зависимости от степени переработки в условиях традиционных технологий производства масложировых продуктов. Впервые получены данные, свидетельствующие о негативном влиянии повторных дезодораций на содержание вредных для здоровья человека веществ - транс-изомеров жирных кислот, ненасыщенных альдегидов и эфиров МХПД, а также на образование потенциально канцерогенных летучих веществ, например 1,2-эпоксибутана. Впервые показана взаимосвязь условий хранения масложировой продукции разной степени очистки (температуры, влажности, освещения и аэрации, близких к нормальным, и длительности хранения) и уровней содержания в ней летучих вторичных продуктов окислительной порчи и технологических контаминантов. Впервые в Российской Федерации проведены исследования содержания технологических контаминантов (эфиров МХПД и глицидиловых эфиров) в пищевых растительных маслах, жирах и масложировых продуктах, представленных на отечественном рынке. Подтвержден факт, что наличие рафинированных дезодорированных масел в составных масложировых продуктах связано с повышенными уровнями эфиров МХПД и глицидиловых эфиров.
На основании полученных данных был сформулирован рейтинг опасности для здоровья человека индивидуальных вторичных продуктов окисления, образующихся и накапливающихся в процессе производства и последующей очистки пищевых растительных масел путем дезодорации, в том числе по нескольким циклам. Была проведена гигиеническая оценка риска потребления населением МХПДЭ и ГЭ, которая выявила необходимость в разработке и внедрении технологий управляемого снижения и предотвращения образования глицидиловых эфиров и глицидола, имеющего статус «вероятного канцерогена человека» и относящегося к группе 2А МАИР, в конечном масложировом продукте.
The book reviews recent developments in the field of nanomaterials science and technology. Topics covered include methods of fabrication of nanomaterials and nanocomposites, and their applications in areas such as Optoelectronics, Cosmetics, Energy Conversion Cells, Soil and Water Treatment, Agricultural Engineering, Food Sciences, Leather Production, and Photocatalysis.
Nanocomposite and their applications
Nanotechnology has been around for many years and has benefited many industries with various applications. Cosmetics and detergents are among the most frequently used customer products blended with nanomaterials. The uniqueness and advantages of nanomaterials along with successful advertisements are keys for the acceptance of these products in the market and households. This chapter summarizes the types of nanomaterials and their roles in cosmetics and detergents. The procedures involved in the synthesis of nanomaterials, product formulations, and the mechanisms of nanoparticles are also highlighted to shed a better understanding on the different functions of nanomaterials. In addition, this chapter also discusses the implications of nanomaterials in these products to human health and environment.
Recent toxicology studies suggest that nanosized aggregates of fullerene molecules can enter cells and alter their functions, and also cross the blood-brain barrier. However, the mechanisms by which fullerenes penetrate and disrupt cell membranes are still poorly understood. Here we use computer simulations to explore the translocation of fullerene clusters through a model lipid membrane and the effect of high fullerene concentrations on membrane properties. The fullerene molecules rapidly aggregate in water but disaggregate after entering the membrane interior. The permeation of a solid-like fullerene aggregate into the lipid bilayer is thermodynamically favoured and occurs on the microsecond timescale. High concentrations of fullerene induce changes in the structural and elastic properties of the lipid bilayer, but these are not large enough to mechanically damage the membrane. Our results suggest that mechanical damage is an unlikely mechanism for membrane disruption and fullerene toxicity.
Fullerene-C60 (C60) is mainly applied in the aqueous phase by wrapping with water-soluble polymer or by water-solublizing chemical-modification, whereas C60 dissolved in oil is scarcely applied; still less explicable is its toxicity.We dissolved C60 in squalane at near-saturated or higher concentrations (220-500 ppm), named LipoFullerene (LF-SQ),and examined its biological safety. LF-SQ was administered at doses of 0.49-1000 microg/ml to fibroblast cells Balb/3T3, and showed that cell viability was almost equal to that of the control regardless of the UVA- or sham-irradiation, indicating no phototoxicity. Reverse mutation by LF-SQ was examined on four histidine-demanding strains of Salmonella typhimurium and a tryptophan-demanding strain of Escherichia coli. As for the dosages of LF-SQ (313-5000 microg/plate), the dose-dependency of the number of reverse mutation colonies of each strain did not show a marked difference when compared with the negative control, regardless of the metabolic activation, in contrast to twice or more differences for five positive controls(sodium azide, N-ethyl-N'-nitro-N-nitrosoguanidine, 2-nitrofluorene, 9-aminoacridine, and 2-aminoanthracene). In human skin biopsy built in a diffusion chamber, C60 permeated into the epidermis at 33.6 nmol/g tissue (24.2 ppm), on administration with LF-SQ containing 223 ppm of C60, but not detected in the dermis even after 24 hrs, as analysed by HPLC. It is presumed that LF-SQ can permeate into the epidermis via the corneum but can not penetrate the basement membrane,and so can not reach into the dermis, suggesting no necessity for considering a toxicity of C60 due to systemic circulation via dermal veins. Thus, C60 dissolved in squalane may not give any significant biological toxic effects such as photocytotoxicity,bacterial reverse mutagenicity, and permeability into the human skin.
Fullerenes represent a group of nanoparticles discovered in 1985. They are spherical molecules consisting entirely of carbon atoms (C(x)) to which side chains can be added, furnishing compounds with widely different properties. Fullerenes interact with biological systems, for example, by enzyme inhibition, causing phototoxic reactions, being scavengers of reactive oxygen species and free radicals, in addition to being able to initiate free radical reactions. Absorption, distribution and excretion strongly depend on the properties of the side chains. The pristine C(60) has a very long biological half-life, whereas the most water-soluble derivatives are eliminated from the exposed animals within weeks. A long biological half-life raises concern about bioaccumulation and long-term effects. In general, the acute oral, dermal and airway toxicity is low. However, few relevant experimental studies of repeated dose toxicity, reproductive toxicity and carcinogenic effect are available. The data suggest that direct DNA damaging effects are low, but formation of reactive oxygen species may cause inflammation and genetic damage. Apparently, it is dose-dependent whether a beneficial or an adverse effect occurs.
The influence of mineral oil, squalane, squalene, or peanut oil on the antitumor activity of emulsified Bacillus Calmette-Guérin cell walls or emulsified trehalose-6,6'-dimycolate was studied in mice, each with an established transplant of a syngeneic fibrosarcoma. Each animal received an intratumoral injection of Bacillus Calmette-Guérin cell walls (0.6 mg/mouse) or trehalose-6,6'-dimycolate (0.1 mg/mouse) emulsified in 1 to 10% oil. Emulsions of squalene or squalane but not peanut oil were effective substitutes for mineral oil as carriers of Bacillus Calmette-Guérin cell walls in the treatment of the tumor. Trehalose-6,6'-dimycolate was therapeutically active when it was incorporated in any of these four oils. The number of animals in which tumor regressed completely depended on the concentration of oil in the emulsion.
Products of lipid peroxidation (malonaldehyde, Schiff-bases) were detected in human skin. These products were increased after UV-light exposition, on chronically sun-exposed areas as well as with advancing age. Malonaldehyde cross linked epidermal glucose-6-phosphate-dehydrogenase and diminished their activity.
In the previous papers, we demonstrated, by using rats, that squalane (2,6,10,15,19, 23-hexamethyltetracosane) could stimulate the fecal excretion of 2,3,4,7,8-pentachlorodibenzofuran, the most important etiologic agent of Yusho, which was accumulated in the body of rat. We also reported that, in rats and dogs, squalane did not show any appreciable toxic signs during 3-month treatment, though a part of squalane was absorbed from gastrointestinal tract of dogs. In the present paper, we have investigated the elimination of absorbed squalane in beagle dogs. During the treatment with squalane orally at a dose of 1200 mg/kg/day for 14 days, the fecal excretion of squalane per day was 65-90% of the daily dose. After the treatment (on the day 14), squalane levels in blood and hair were about 30 ppm and 14640 ppm, respectively. On the day 56 after the first dosing, squalane was not detected in blood. On the day 70, squalane level in hair was reduced to about 1% of that on the day 14. Squalane levels in skin, liver, adipose tissue and small intestine on the day 70 were also reduced compared with that on the day 42. Moreover, small amount of squalane was still excreted into feces from the day 15 to the day 70. These results suggested that absorbed squalane was gradually excreted through feces and skin in dogs.
In the previous papers, we demonstrated, by using rats, that squalane (2,6,10,15,19,23-hexamethyltetracosane) could stimulate the fecal excretion of 2,3,4,7,8-pentachlorodibenzofuran, which was regarded as the most important etiologic agent of yusho among PCB and PCDF congeners found in the causal rice oil. We also reported that, in rats, squalane was not essentially absorbed from the gastrointestinal tract, and did not show any appreciable side effects during the 3-month treatment. In the present paper, we have investigated the distribution, excretion and subacute toxicity of squalane in beagle dogs. The fecal excretion of squalane accounted for about 83% of dose during the initial 2 days after administration at a single oral dose of 1,200 mg/kg to male dogs. On day 3, absorbed squalane was mostly distributed to the hair and the skin, and the concentrations in these tissues were decreased on day 6. These results suggested that most of squalane administered orally was not absorbed from the gastrointestinal tract, but a part was absorbed and excreted through the hair. In addition, squalane distributed into the liver was found to be eliminated rather slowly. A long-term (13-week) treatments with squalane orally at doses of 400 mg/kg/day or 1,200 mg/kg/day in male and female dogs, resulted also in accumulation of squalane in the liver at a level of about 3% (400 mg/kg) or about 6% (1,200 mg/kg) of the daily dose. This accumulation of squalane in the liver was highest among all the tissues. Nevertheless, no appreciable toxic signs were observed in the serum biochemical tests and the hepatic functional test for squalane groups. Therefore, squalane accumulating in the liver, did not seem to disturb the hepatic physiological functions. It was suggested also in a long-term treatment that the skin and the hair played the most important role in the elimination of squalane. In conclusion, the present studies on subacute toxicity tests suggested that squalane did not give any significant toxic effects on dogs as well as rats.
The lethal effects of linoleic acid and its hydroperoxide on human diploid fibroblasts were quite similar, and that of the reaction mixture from the autoxidation of the hydroperoxide was considerable. Some unsaturated aliphatic aldehydes, the secondary products of the hydroperoxide autoxidation, were identified and their toxicity toward the cells was examined. Among them, (E,E)-2,4-nonadienal, (E,E)-2,4-decadienal, and (E)-4-hydroxy-2-nonenal were the most toxic; e.g., in the presence of 25 microM nonadienal, decadienal, or hydroxynonenal, 75, 90, or almost 100% of the cells, respectively, underwent lysis within one day. Generally, alkenals were toxic and alkanals non-toxic. The toxicity was enhanced as the number of double bonds in each molecule was increased and also as the carbon chain was lengthened.
Freeze cracking methods for the preparation of scanning electron microscope specimens are described and the results obtained in rat kidney and human spleen are demonstrated. Fixed tissue pieces immersed either in ethanol, isoamyl acetate, or in 40% dimethyl sulfoxide were quench frozen in liquid nitrogen or in Freon 22 cooled by liquid nitrogen and cracked. Freeze cracking in ethanol and isoamyl acetate produced clean and flat fracture surfaces causing excellent visualization of the lining surfaces of the opened vessels, tubules and tissue spaces. Freeze cracking in dimethyl sulfoxide tended to cause fracture along cell surfaces and intracellular membranes.
Gelatin capsules containing squalane partially purified bone morphogenetic protein (BMP) complex were placed on the perimuscular membrane of rats. Two kinds of control, gelatin capsules containing only BMP and those bearing squalane only, were used. The embedded areas were histopathologically examined at 3 and 6 wk after the operation. The observations revealed that the squalane/BMP complex elicited wide heterotopic bone formation with bone marrow tissue, suggesting that squalane is a possible carrier of BMP for clinical applications.