Fermented tea leave extract against oxidative stress and ageing of skin in vitro and in vivo.

OBJECTIVE: The objective is to develop a natural and stable anti-oxidative stress and anti-ageing ingredient. In this study, we evaluated the changes in white tea leaves fermented with Eurotium cristatum PLT-PE and Saccharomyces boulardii PLT-HZ and their efficacy against skin oxidative stress. METHODS: We employed untargeted metabolomics technology to analyse the differential metabolites between tea extract (TE) and fermented tea extract (FTE). In vitro, using H2O2-induced HaCaT cells, we evaluated cell vitality, ROS, and inflammatory factors (TNF-α, IL-1ß, and IL-6). Additionally, we verified the effects on the extracellular matrix and nuclear DNA using fibroblasts or reconstructed skin models. We measured skin hydration, elasticity, wrinkle area, wrinkle area ratio, erythema area, and erythema area ratio in volunteers after using an emulsion containing 3% FTE for 28 and 56 days. RESULTS: Targeted metabolomics analysis of white tea leaves yielded more than 20 differential metabolites with antioxidant and anti-inflammatory activities, including amino acids, polypeptides, quercetin, and liquiritin post-fermentation. FTE, compared to TE, can significantly reduce reactive oxygen species (ROS) and protect against oxidative stress-induced skin damage in H2O2-induced HaCaT cells. FTE can inhibit H2O2-induced collagen degradation by suppressing the MAPK/c-Jun signalling pathway and can also mitigate the reactive oxygen species damage to nuclear DNA. Clinical studies showed that the volunteers' stratum corneum water content, skin elasticity, wrinkle area, wrinkle area ratio, erythema area, and erythema area ratio significantly improved from the baseline after 28 and 56 days of FTE use. CONCLUSION: This study contributes to the growing body of literature supporting the protective effects against skin oxidative stress and ageing from fermented plant extracts. Moreover, our findings might inspire multidisciplinary efforts to investigate new fermentation techniques that could produce even more potent anti-ageing solutions.

OBJECTIF: L'objectif est de développer un ingrédient naturel et stable contre le stress oxydatif et anti­âge. Dans cette étude, nous avons évalué les modifications dans les feuilles de thé blanc fermentées avec la PLT­PE Eurotium cristatum et la PLT­HZ Saccharomyces boulardii et leur efficacité contre le stress oxydatif cutané. MÉTHODES: Nous avons utilisé une technologie de métabolomique non ciblée pour analyser les métabolites différentiels entre l'extrait de thé (ET) et l'extrait de thé fermenté (ETF). In vitro, à l'aide de cellules HaCaT induites par l'H2O2, nous avons évalué la vitalité cellulaire, les ERO et les facteurs inflammatoires (TNF­α, IL­1ß, and IL­6). Nous avons également vérifié les effets sur la matrice extracellulaire et l'ADN nucléaire à l'aide de fibroblastes ou de modèles cutanés reconstruits. Nous avons mesuré l'hydratation de la peau, l'élasticité, la surface de rides, le rapport des surfaces de rides, la surface d'érythème, et le rapport des surfaces d'érythème chez des volontaires ayant utilisé une émulsion contenant 3% d'ETF pendant 28 et 56 jours. RÉSULTATS: L'analyse métabolomique ciblée des feuilles de thé blanc a révélé plus de 20 métabolites différentiels ayant des activités antioxydantes et anti­inflammatoires, notamment des acides aminés, des polypeptides, de la quercétine et de la liquiritine après fermentation. Par rapport à l'ET, l'ETF peut réduire significativement les espèces réactives de l'oxygène (ERO) et protéger contre les lésions cutanées induites par le stress oxydatif dans les cellules HaCaT induites par l'H2O2. L'ETF peut inhiber la dégradation du collagène induite par l'H2O2 en supprimant la voie de signalization MAPK/c­Jun et peut également atténuer les dommages causés par les espèces réactives de l'oxygène à l'ADN nucléaire. Les études cliniques ont montré que la teneur en eau de la couche cornée des volontaires, l'élasticité de la peau, la surface de rides, le rapport des surfaces de rides, la surface d'érythème et le rapport des surfaces d'érythème se sont significativement améliorés par rapport à la référence après 28 et 56 jours d'utilisation d'ETF. CONCLUSION: Cette étude contribue au corpus croissant de littérature soutenant les effets protecteurs des extraits de plantes fermentées contre le stress oxydatif cutané et le vieillissement. En outre, nos résultats pourraient inspirer des efforts pluridisciplinaires pour étudier de nouvelles techniques de fermentation susceptibles de produire des solutions anti­âge encore plus puissantes.

Enzymatically modified isoquercitrin and its protective effects against photoaging: In-vitro and clinical studies.

This research examines the anti-aging potential of the flavonoid derivative of isoquercitrin known as enzymatically modified isoquercitrin (EMIQ). Initial HPLC analyses showed that EMIQ used in the study contained 1-12 glucosides and 10.7% pentahydroxyflavonoids, promising potent antioxidant properties. In subsequent in-vitro studies with UVA-exposed human dermal fibroblasts (HDFa), EMIQ demonstrated protective properties by reducing collagen damage. It modulated both the TGFß/Smad pathway and the MMP1 pathway, contributing to collagen preservation. This protective effect was further confirmed using the T-Skin™ model, a reconstructed full-thickness human skin model, which illustrated that EMIQ could defend the physiological structures of both the epidermis and dermis against UV radiation. A 28-day clinical trial with 30 volunteers aged 31-55 years highlighted EMIQ's effectiveness. Participants using EMIQ-containing Essence displayed reduced facial trans-epidermal water loss and skin roughness, alongside improved skin elasticity. This study emphasizes EMIQ's potential as an anti-photoaging ingredient in cosmetics, warranting further research. The findings pave the way for developing innovative skincare products addressing photoaging effects.

Spectrophotometric evaluation of color errors generated in the visual color duplication procedure for current ceramic veneers.

BACKGROUND/PURPOSE: Color errors associated with the visual color duplication approach for ceramic laminate veneers are still challenging in esthetic dentistry. The aim of this study is to evaluate color errors generated during traditional visual shade matching approach. MATERIALS AND METHODS: Eighteen stooth-shaped veneer discs (shade A2 and 0.7 mm in thickness) were fabricated using six veneer materials. The veneer specimens placed on five extracted teeth with nominal shade A2 formed veneer-tooth combinations. Color coordinates of the A2 shade tab, the extracted teeth, and the veneer-tooth combinations were measured using a spectrophotometer. Then, the veneers were reduced to 0.5 mm, and 0.3 mm in thickness consecutively. Color measurements were performed repeatedly. Color differences of the extracted teeth to veneer-tooth combinations (ΔEt-v), veneer-tooth combinations to shade tab (ΔEv-s), and translucency parameter (TP) values were calculated and analyzed using Two-way ANOVA. RESULTS: ΔEt-v ranged from 2.0937 to 5.0603 (mean of 3.1833±1.5485). Mean of ΔEv-s was 4.0103±1.8508. ΔEt-v and ΔEv-s values were significantly influenced by veneer material and thickness (P<0.05). TP values decreased gradually with the lessening of veneers thickness. CONCLUSION: Acceptable color duplication of ceramic veneers cannot be achieved by routine visual shade replica protocols, when the thickness of veneers is less than 0.7 mm. Specified color matching standards for the ceramic veneers are needed.

* Coculture with Partial Digestion Notochordal Cell-Rich Nucleus Pulposus Tissue Activates Degenerative Human Nucleus Pulposus Cells.

Recent studies suggested that notochordal cells (NCs) and NC-conditioned medium (NCCM) can stimulate cell viability and matrix production of nucleus pulposus cells (NPCs). However, the potential of notochordal cell-rich nucleus pulposus (NRNP) incorporating the native environment of the intervertebral disc (IVD) has not been evaluated. The objective of this study was to develop an optimal NRNP model and test whether it can allow a significant level of NPC activation in vitro. Rabbit NRNP explants were divided into three groups according to different digestion time: digestion NRNP of 8 h, partial digestion NRNP of 2 h, and natural NRNP. Cell viability and NC phenotype were compared between these groups after 14 days of incubation. The products of the selected partial digestion NRNP group were then cocultured with human degenerated NPCs for 14 days. NPC viability, cell proliferation and senescence, the production of glycosaminoglycan (GAG) found in extracellular matrix, and NP matrix production by NPCs were assessed. The results showed that coculturing with partial digestion NRNP significantly improved the cell proliferation, cell senescence, and disc matrix gene expression of NPCs compared with those in the monoculture group. In addition, GAG/DNA ratio in the coculture group increased significantly, while the level of collagen II protein remained unchanged. In this study, we demonstrated that partial digestion NRNP may show a promising potential for NPC regeneration in IVD tissue engineering.

BMP7-Based Functionalized Self-Assembling Peptides Protect Nucleus Pulposus-Derived Stem Cells From Apoptosis In Vitro.

Tissue engineering has shown great success in the treatment of intervertebral disk degeneration (IVDD) in the past decade. However, the adverse and harsh microenvironment associated in the intervertebral disks remains a great obstacle for the survival of transplanted cells. Although increasing numbers of new materials have been created or modified to overcome this hurdle, a new effective strategy of biological therapy is still required. In this study, bone morphogenic protein 7 (BMP7)-based functionalized self-assembling peptides were developed by conjugating a bioactive motif from BMP-7 (RKPS) onto the C-terminal of the peptide RADARADARADARADA (RADA16-I) at a ratio of 1:1 to form a new RADARKPS peptide. Human nucleus pulposus-derived stem cells (NPDCs) were cultured in the presence of RADA-RKPS or RADA16-I in an apoptosis-promoting environment that was induced by tumor necrosis factor-alpha, and cells were cultured with RADA16-I in normal medium that served as the control group. After 48 h of apoptosis induction, the viability, proliferation, apoptosis rate, and expression of apoptosis-related genes of NPDCs in the different groups were evaluated, and the differentiation of NPDCs toward nucleus pulposus-like cells was tested. The results showed that the RADA-RKPS peptide could significantly protect the survival and proliferation of NPDCs. In addition, the application of RADA-RKPS decreased the rate of cell apoptosis, as detected by TUNEL-positive staining. Furthermore, our in vitro study confirmed the apoptosis-protecting effects of RADA-RKPS peptides, which significantly reduced the BAX/BCL-2 ratio of NPDCs and upregulated the gene expression of collagen II a1, aggrecan, and Sox-9 after 48 h of apoptosis induction. Collectively, these lines of evidence suggest that RADA-RKPS peptides confer a protective effect to NPDCs in an apoptosis environment, suggesting their potential application in the development of new biological treatment strategies for IVDD.

Biological evaluation of human degenerated nucleus pulposus cells in functionalized self-assembling peptide nanofiber hydrogel scaffold.

Nucleus pulposus (NP) tissue engineering has been proposed as a novel biological treatment for early-stage intervertebral disc degeneration. In this study, a novel functional self-assembling peptide PKP was first designed by linking the short functional motif of bone morphogenetic protein-7 (BMP7) to the C-terminal of RADA16-I, and another new functional self-assembling peptide was obtained by mixing RKP with RADA16-I. Then, the biocompatibilities and bioactivities of RKP and RAD-RKP for human degenerated nucleus pulposus cells (hNPCs) were studied in vitro. Atomic force microscopy and scanning electron microscopy (SEM) confirmed that both RKP and RAD-RKP could self-assemble into three-dimensional (3D) nanofiber hydrogel scaffolds in a culture medium at 37°C. After the hNPCs were cultured in 3D scaffolds, both RKP and RAD-RKP exhibited reliable attachment and extremely low cytotoxicities (<14%), which were verified by SEM and cytotoxity assays, respectively. Our results also showed that the functional-based scaffolds could increase the proliferation and migration of hNPCs after 7 days compared with culture plates and pure RADA16-I. Quantitative real-time polymerase chain reaction demonstrated that the expressions of collagen II α1, Sox-9, and aggrecan were upregulated, while collagen I α1 was downregulated by functional-based scaffolds after 28 days. Furthermore, we also confirmed that RAD-RKP exhibited a higher hNPC proliferation, migration, and expression of Sox-9 and aggrecan compared with pure RKP. Therefore, the results of this study indicated that the BMP7 short motif-designed functional self-assembling peptide nanofiber hydrogels could be used as excellent scaffolds in NP tissue engineering, and RAD-RKP might have further potential application in human mild degenerated NP tissue regeneration.

[The construction of recombinant adenovirus containing p38MAPK gene and its expression in mouse osteoblast-like cells (MC3T3-E1)].

AIM: To construct a recombinant adenovirus vector containing p38MAPK gene and to identify its expression in mouse osteoblast-like cells, MC3T3-E1, in vitro. METHODS: The p38MAPK gene was amplified by PCR from mouse liver cDNA library, and inserted into pMD18-T vector and sequenced. Double digested with Bgl II and Hind III, p38MAPK gene was inserted into pShuttle-CMV. This recombinant plasmid was linearized by Pme I and electronically transfected into BJ5183 to get the recombinant adenovirus vector Ad-p38MAPK. Then the Ad-p38MAPK was obtained by packaging Pac I linearized in AD293 cells. The recombinant adenovirus expressing p38MAPK was infected into osteoblast- like cells(MC3T3-E1).The expression of exogenous p38MAPK was observed by Western blot. RESULTS: The recombinant plasmid Ad-p38MAPK was successfully generated, which increased p38MAPK protein expression levels in MC3T3-E1. CONCLUSION: The bioactive recombinant adenovirus Ad-p38MAPK has been successfully constructed. The study laid a foundation for further research of the function of p38MAPK in osteoblast.

Kordiimonas lacus sp. nov., isolated from a ballast water tank, and emended description of the genus Kordiimonas.

A Gram-negative, motile, rod-shaped bacterial strain, designated S3-22(T), was isolated from a sediment sample collected from a ballast water tank of a commercial ship and subjected to a polyphasic taxonomic characterization. The isolate formed small, light-yellow, semi-translucent and circular colonies on solid complex media. The strain was oxidase- and catalase-positive and metabolized a large number of carbon sources. Chemotaxonomic analysis showed ubiquinone Q-10 as predominant respiratory quinone, phosphatidylglycerol and an unidentified glycolipid as major polar lipids and iso-C(17 : 1)ω9c, iso-C(15 : 0), C(16 : 1)ω7c and/or iso-C(15 : 0) 2-OH, C(16 : 0), iso-C(17 : 0) and C(18 : 1)ω7c as major fatty acids and the hydroxy fatty acids iso-C(17 : 0) 3-OH and C(16 : 0) 3-OH. The genomic DNA G+C content was 54.9 mol%. 16S rRNA gene sequence analysis revealed that the isolate has 96.1 % similarity to the type strain of Kordiimonas gwangyangensis, the sole described species within the order Kordiimonadales, and less than 91.0 % similarity to other recognized species. On the basis of phenotypic and genotypic data, strain S3-22(T) represents a novel species of the genus Kordiimonas, for which the name Kordiimonas lacus sp. nov. is proposed, with the type strain S3-22(T) (=CGMCC 1.9109(T) =JCM 16261(T)). An emended description of the genus Kordiimonas is also presented.

[Study on recovery of ethyl acetate after extracting tripterygium wifordii extractum with ethyl acetate].

OBJECTIVE: To increase the recovery rate of ethyl acetate after extracting tripterygium wifordii extractum and to decrease product cost. METHOD: After extracting tripterygium wifordii extractum with ethyl acetate, 3 times saturated salt water was added in it so as to recovery ethyl acetate distilled under normal atmospheric pressure. Ethyl acetate containing salt water was purified through Na2SO4 column. RESULT: Ethyl acetate purified could be used repeatedly and the recovery rate was up to 85%. CONCLUSION: This method is completely adapted for mass production.