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13 pages, 911 KiB  
Article
Immune-Related Gene Expression Responses to In Ovo Stimulation and LPS Challenge in Two Distinct Chicken Genotypes
by Anna Slawinska, Aleksandra Dunisławska, Artur Kowalczyk, Ewa Łukaszewicz and Maria Siwek
Genes 2024, 15(12), 1585; https://doi.org/10.3390/genes15121585 (registering DOI) - 9 Dec 2024
Abstract
Background: In ovo stimulation introduces bioactive compounds, such as prebiotics, probiotics, or synbiotics into incubating eggs to enhance gut health and immune system development in chickens. This study aimed to determine the genetic and environmental effects modulating responses to in ovo stimulation in [...] Read more.
Background: In ovo stimulation introduces bioactive compounds, such as prebiotics, probiotics, or synbiotics into incubating eggs to enhance gut health and immune system development in chickens. This study aimed to determine the genetic and environmental effects modulating responses to in ovo stimulation in commercial broilers and Green-legged Partridge-like (GP) native chickens. Methods: Eggs were stimulated on day 12 of incubation with prebiotics (GOS—galactooligosaccharides), probiotics (Lactococcus lactis subsp. cremoris), or synbiotics (GOS + L. lactis), with controls being mock-injected. Hatched chicks were reared in group pens and challenged with lipopolysaccharide (LPS) on day 42 post-hatching. Cecal tonsils (CT) and spleens were harvested 2 h post-challenge. RT-qPCR was used to analyze the relative gene expression of cytokine genes: IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12p40, and IL-17. Results: The results show that genotype influenced the expression of all immune-related genes, with broiler chickens exhibiting stronger innate inflammatory responses than native chickens. LPS induced both mucosal (CT) and systemic (spleen) immune responses in broilers but only systemic (spleen) responses in native chickens. Conclusions: In ovo stimulation had less of an impact on cytokine gene expression than LPS challenge. Broilers expressed higher inflammatory immune responses than GP native chickens. Full article
(This article belongs to the Special Issue Poultry Genetics and Genomics—2nd Edition)
14 pages, 7233 KiB  
Article
Non-Targeted Metabolomics Analysis Reveals Metabolite Profiles Change During Whey Fermentation with Kluyveromyces marxianus
by Yansong Gao, Lei Gao, You Kang, Ge Yang, Zijian Zhao, Yujuan Zhao and Shengyu Li
Metabolites 2024, 14(12), 694; https://doi.org/10.3390/metabo14120694 (registering DOI) - 9 Dec 2024
Abstract
Background: Whey fermentation could produce bioactive substances with immunomodulatory effects, metabolic syndrome modulation, and antioxidant properties, thereby imparting functional characteristics to products and facilitating the development of novel foods with health-promoting potential. Methods: A non-targeted metabolomics approach using liquid chromatography–mass spectrometry (LC-MS) was [...] Read more.
Background: Whey fermentation could produce bioactive substances with immunomodulatory effects, metabolic syndrome modulation, and antioxidant properties, thereby imparting functional characteristics to products and facilitating the development of novel foods with health-promoting potential. Methods: A non-targeted metabolomics approach using liquid chromatography–mass spectrometry (LC-MS) was employed to investigate changes in the metabolite profiles of whey fermented by Kluyveromyces marxianus strain KM812 over varying fermentation durations. Results: The findings demonstrated a progressive enrichment of metabolites over time. A total of 151 differential metabolites were identified and categorized primarily into amino acids, peptides, and analogues, fatty acids and conjugates, and carbohydrates and conjugates, as well as benzoic acids and derivatives. The highest relative content of whey metabolites was observed at 48 h of fermentation, with a cumulative increase of 1.45-fold, 1.49-fold, 3.39-fold, and 1.24-fold for peptides and amino acids, peptides, and analogues, fatty acids and conjugates, and carbohydrates and conjugates, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed associations with 23 specific metabolites and delineated 9 metabolic pathways, predominantly involved in amino acid and lipid metabolism. Conclusions: Based on the above, KM812 could effectively degrade macromolecular substances in whey into small molecules such as L-isoleucine, ornithine, betaine, α-linolenic acid, and palmitoleic acid, thereby influencing the nutritional and functional properties of whey. In-depth analysis of the metabolic products in KM812-fermented whey could provide a theoretical basis for the development of functional foods derived from small molecules in the future. Full article
(This article belongs to the Section Food Metabolomics)
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<p>(<b>A</b>,<b>B</b>) PBC of ESI+ and ESI− mode in whey at different fermentation times.</p>
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<p>(<b>A</b>−<b>J</b>) PCA score plots (<b>A</b>,<b>B</b>), PLS−DA score plots (<b>C</b>,<b>D</b>), OPLS−DA score plots (<b>E</b>,<b>F</b>), accompanied by permutation tests (<b>G</b>,<b>H</b>), and charge ratio and <span class="html-italic">p</span> value scatter plots (<b>I</b>,<b>J</b>) of whey at different fermentation times in the ESI + mode and in the ESI− mode, respectively (each point in (<b>I</b>,<b>J</b>) denotes one metabolite, with the horizontal coordinates denoting the mass−to−charge ratios of the tertiary substances; the vertical coordinates denoting the <span class="html-italic">p</span>−values of the −log10 of the <span class="html-italic">p</span> value. Larger values of the vertical coordinate indicate more significant differential expression. Red dots represent up−regulated differentially expressed metabolites, grey dots represent metabolites that did not satisfy the filtering parameters, and the size of the dots indicates the VIP value).</p>
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<p>Hierarchical clustering heatmap of differential metabolites of fermented whey at different stages of fermentation (the magnitude of the relative content in the plot is shown by differences in colour, with redder colours showing higher expression and bluer colours showing lower expression).</p>
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<p>(<b>A</b>,<b>B</b>) Venn diagram of differential metabolites in intergroup comparisons (<b>A</b>); histogram of differential metabolite statistics for group comparisons (<b>B</b>) (X−axis indicates number of differential metabolites and Y−axis indicates group comparison conditions).</p>
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<p>(<b>A</b>–<b>I</b>) Molecular expression pattern plot (each subplot represents a different cluster, with the x−axis indicating fermentation time and the y−axis representing the average expression level of the molecule within each group. Cluster1 Includes 17 metabolites (<b>A</b>); Cluster2 Includes 17 metabolites (<b>B</b>); Cluster3 Includes 7 metabolites (<b>C</b>); Cluster4 Includes 12 metabolites (<b>D</b>); Cluster5 Includes 28 metabolites (<b>E</b>); Cluster6 Includes 23 metabolites (<b>F</b>); Cluster7 Includes 22 metabolites (<b>G</b>); Cluster8 Includes 7 metabolites (<b>H</b>); Cluster1 Includes 18 metabolites (<b>I</b>);).</p>
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<p>Metabolic pathway impact factor bubble diagram. (Each point in the diagram represents a metabolic pathway, the horizontal coordinates are the Impact values enriched into different metabolic pathways, and the vertical coordinates are the enriched pathways. The dots indicate the corresponding number of metabolic molecules in the pathway. The colours correlate with the <span class="html-italic">p</span>-value; the redder the colour, the smaller the <span class="html-italic">p</span>-value, and the bluer the colour, the larger the <span class="html-italic">p</span>-value.)</p>
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<p>Metabolic network diagram between specific metabolites and KEGG pathways (red markers represent specific metabolites, different coloured rounded rectangles represent different pathways enriched by KEGG analysis).</p>
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37 pages, 5914 KiB  
Review
Integrating Physical and Biochemical Cues for Muscle Engineering: Scaffolds and Graft Durability
by Farbod Yousefi, Lauren Ann Foster, Omar A. Selim and Chunfeng Zhao
Bioengineering 2024, 11(12), 1245; https://doi.org/10.3390/bioengineering11121245 - 9 Dec 2024
Viewed by 101
Abstract
Muscle stem cells (MuSCs) are essential for skeletal muscle regeneration, influenced by a complex interplay of mechanical, biochemical, and molecular cues. Properties of the extracellular matrix (ECM) such as stiffness and alignment guide stem cell fate through mechanosensitive pathways, where forces like shear [...] Read more.
Muscle stem cells (MuSCs) are essential for skeletal muscle regeneration, influenced by a complex interplay of mechanical, biochemical, and molecular cues. Properties of the extracellular matrix (ECM) such as stiffness and alignment guide stem cell fate through mechanosensitive pathways, where forces like shear stress translate into biochemical signals, affecting cell behavior. Aging introduces senescence which disrupts the MuSC niche, leading to reduced regenerative capacity via epigenetic alterations and metabolic shifts. Transplantation further challenges MuSC viability, often resulting in fibrosis driven by dysregulated fibro-adipogenic progenitors (FAPs). Addressing these issues, scaffold designs integrated with pharmacotherapy emulate ECM environments, providing cues that enhance graft functionality and endurance. These scaffolds facilitate the synergy between mechanotransduction and intracellular signaling, optimizing MuSC proliferation and differentiation. Innovations utilizing human pluripotent stem cell-derived myogenic progenitors and exosome-mediated delivery exploit bioactive properties for targeted repair. Additionally, 3D-printed and electrospun scaffolds with adjustable biomechanical traits tackle scalability in treating volumetric muscle loss. Advanced techniques like single-cell RNA sequencing and high-resolution imaging unravel muscle repair mechanisms, offering precise mapping of cellular interactions. Collectively, this interdisciplinary approach fortifies tissue graft durability and MuSC maintenance, propelling therapeutic strategies for muscle injuries and degenerative diseases. Full article
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<p>Understanding muscle stem cell states and PAX7 distribution. Understanding Muscle Stem Cell States: Quiescence, Activation, Proliferation, Differentiation, and Self-Renewal. Panel (<b>a</b>) illustrates the dynamic transition of muscle stem cells through various states, starting from quiescence, progressing through activation and proliferation, leading to differentiation, and the potential for self-renewal [<a href="#B29-bioengineering-11-01245" class="html-bibr">29</a>]. The schematic emphasizes the directional flow and sequential nature of these states, with each phase contributing to muscle regeneration and maintenance. Panel (<b>b</b>) presents PAX7 distribution curves within the niche, highlighting the variability and shifts in PAX7 expression levels during different states. The distribution curves illustrate how PAX7 expression peaks and shifts as muscle stem cells transition between quiescence, activation, and other states, reflecting their changing roles and regulatory mechanisms within the cellular environment [<a href="#B35-bioengineering-11-01245" class="html-bibr">35</a>]. Together, these panels provide a comprehensive overview of muscle stem cell state transitions and the critical role of PAX7 as a marker modulating these processes.</p>
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<p>Cellular interactions and regulatory mechanisms in the muscle stem cell niche. This figure depicts the complex interplay among various cellular components of the muscle stem cell niche, including fibro-adipogenic progenitors (FAPs), pericytes, fibroblasts, and macrophages (M1 and M2 phenotypes). The top section illustrates the role of macrophages, showing the transition from pro-inflammatory M1 macrophages, which secrete cytokines like TNF-α and IL-1β, to anti-inflammatory M2 macrophages involved in tissue repair and regeneration, producing factors such as IL-10 and TGF-β. Adjacent panels highlight interactions with T cells and their influence on the local inflammatory response. The central portion emphasizes cellular and extracellular matrix (ECM) interactions mediated by FAPs, pericytes, and fibroblasts, influencing the structural and regenerative dynamics of the muscle niche. The figure also outlines cell cycle regulation, detailing checkpoints and the role of cyclins (Cyclin A, B, D, E) and cyclin-dependent kinases (CDKs) in governing progression through G1, S, G2, and M phases. The accompanying graph on cyclin and CDK concentration during the cell cycle phases provides a visual representation of how cyclin–CDK complexes fluctuate, coordinating MuSC proliferation and differentiation [<a href="#B47-bioengineering-11-01245" class="html-bibr">47</a>]. Specific cyclin–CDK complexes’ elevated levels promote cell cycle progression, while their inhibition leads to cell cycle arrest, thus tightly regulating MuSC proliferation within the niche [<a href="#B48-bioengineering-11-01245" class="html-bibr">48</a>]. Together, these elements underscore the regulatory pathways and cellular interactions that maintain muscle homeostasis and regeneration within the niche.</p>
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<p>Heatmap of molecular pathways influencing muscle stem cell fates. This heatmap provides a visualization of the correlation between various molecular pathways and muscle stem cell fates, based on a qualitative exploratory review of relevant literature. The pathways considered include IL-6/JAK/STAT, TNF-α/NF-κB, TGF-β/SMAD, Notch, ROS/antioxidant mechanisms, and others. Muscle stem cell fates<tt>—</tt>quiescence, activation, proliferation, differentiation, and self-renewal<tt>—</tt>are represented along the <span class="html-italic">x</span>-axis, while molecular pathways are listed on the <span class="html-italic">y</span>-axis. The color gradient reflects the influence score of each pathway, with red hues indicating a positive correlation and blue hues representing a negative correlation. This visualization highlights how different pathways modulate specific cellular states, providing insight into the complex regulatory networks governing muscle stem cell behavior. The heatmap underscores the critical interplay of signaling cascades that dictate stem cell responses and tissue regeneration, as referenced in <a href="#bioengineering-11-01245-t001" class="html-table">Table 1</a>.</p>
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<p>Scaffold characteristics. The figure provides an overview of critical scaffold properties that influence muscle stem cell function and tissue engineering outcomes. The properties are categorized into structural and functional aspects, including vascularity, architecture, pore size, degradation, mechanical properties, and nano-specific modifications. The upper left panels illustrate how cells interact with scaffolds, emphasizing effects on cellular organization, polarity, and matrix binding, driven by diffusion gradients and cell–extracellular matrix (ECM) interactions. The central portion of the figure demonstrates mechanical properties, ranging from viscoelasticity (depicting a strain-stress curve) to stiffness levels (soft to stiff matrices) that provide mechanical cues to cells, critical for their behavior and differentiation. Adjacent panels depict architectural considerations such as pore size and overall scaffold structure, influencing cell infiltration, tissue integration, and mass transport. Degradation profiles are shown, emphasizing scaffold dissolution kinetics that match tissue regeneration rates. On the right side, nano-specific modifications, including particles (solid lipid, polymeric vesicles, quantum dots, etc.), highlight applications such as drug delivery and enhanced cellular interactions. The bottom panels explore complex architectures, like double-network gels that combine rigid and flexible networks, enhancing mechanical resilience and adaptability in dynamic tissue environments. The integration of nanoparticles with radioconjugates illustrates potential therapeutic and imaging applications in regenerative medicine and diagnostics. The progressive narrative of the figure emphasizes the interplay of scaffold properties with biological, mechanical, and nanoscale features for optimizing tissue regeneration outcomes.</p>
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<p>Mechanisms and pathways in muscle stem cell regulation. This figure encompasses multiple schematic representations that outline critical regulatory mechanisms influencing muscle stem cell behavior and fate. Panel (<b>a</b>) illustrates major signaling pathways, including Wnt, BMP, TGF-β, Notch, and PI3K/AKT pathways, emphasizing the journey from receptor activation at the cell membrane to downstream nuclear transcription factors that modulate gene expression, impacting muscle stem cell states and responses. Panel (<b>b</b>) provides detailed pathway diagrams, showcasing interactions among key regulatory molecules in muscle stem cell signaling. This includes cross-talk between different pathways that guide cell proliferation, differentiation, quiescence, and self-renewal, depicting an integrated regulatory network. Panel (<b>c</b>) highlights the role of mechanical forces, such as shear stress and tensile stress, in triggering cellular mechanotransduction processes, with a specific focus on mechanical receptors and their influence on intracellular pathways. Panel (<b>d</b>) illustrates epigenetic regulatory mechanisms, including DNA methylation, histone modification, and non-coding RNAs, which fine-tune muscle stem cell states by modifying chromatin structure and gene expression, contributing to the maintenance of stem cell quiescence, activation, or differentiation. Together, these panels underscore the complexity and integration of biochemical, mechanical, and epigenetic influences on muscle stem cell regulation.</p>
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<p>Network diagram of pharmacotherapy and muscle stem cell pathways. This figure visualizes the integration of pharmacotherapy interventions, such as growth factors, small molecules, and other modulators, with muscle stem cell pathways and markers. Nodes represent key elements of pharmacotherapy and stem cell signaling, while edges illustrate their interactions based on a qualitative exploratory review of the relevant literature. The color-coded nodes indicate distinct muscle stem cell states influenced by pharmacotherapy: blue for quiescence maintenance, green for activation, yellow for proliferation, red for differentiation, and purple for self-renewal. The network emphasizes the complex, multi-faceted approach to muscle regeneration, showcasing how targeted pharmacotherapy influences specific pathways, such as Wnt signaling modulators, SIRT1 activators, BMP signaling, and more. This comprehensive view of pharmacological modulation highlights the interplay between small molecules, growth factors, and muscle stem cell regulatory networks, facilitating targeted therapeutic strategies for muscle repair and regeneration, as referenced in <a href="#bioengineering-11-01245-t003" class="html-table">Table 3</a>.</p>
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<p>Heatmap of scaffold characteristics impact on muscle stem cell markers. This heatmap visually represents the estimated impact of various scaffold characteristics on muscle stem cell markers, based on a non-systematic literature review. The scaffold properties examined include stiffness, porosity, mechanical loading, alignment, and others. Muscle stem cell markers (e.g., Pax7, Myf5, Myod1, Myogenin) are displayed along the <span class="html-italic">x</span>-axis, while scaffold characteristics are on the <span class="html-italic">y</span>-axis. Color intensity and direction indicate the correlation strength, with red hues representing positive correlations and blue hues showing negative correlations. These interactions reflect the influence of scaffold design on stem cell behaviors such as quiescence, activation, proliferation, differentiation, and self-renewal. The heatmap provides insights into how scaffold properties can be strategically modulated to optimize muscle regeneration and stem cell function, summarizing findings based on references listed in <a href="#bioengineering-11-01245-t004" class="html-table">Table 4</a>.</p>
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<p>Network mapping of scaffold properties and muscle stem cell markers. This figure illustrates the estimated relationships between various scaffold properties (e.g., stiffness, alignment, porosity) and muscle stem cell markers, including Pax7, Myf5, Myod1, Myogenin, and others. The network displays interactions based on literature-derived estimates linking scaffold features to different stem cell fates such as quiescence, activation, proliferation, differentiation, and self-renewal. Scaffold properties, represented as nodes, interact through specific pathways with muscle stem cell markers and their regulatory networks. Color-coded nodes indicate stem cell fates influenced by scaffold design: blue represents quiescence, green denotes activation, yellow signifies proliferation, red indicates differentiation, and purple corresponds to self-renewal. The pathways connecting these nodes highlight the mechanistic role of scaffold properties in modulating muscle stem cell behavior and fate, reflecting how physical cues from scaffolds translate into cellular responses. This diagram emphasizes the complexity and interconnectedness of scaffold-induced regulation, illustrating their critical influence on stem cell signaling and tissue regeneration dynamics. The network is informed by selective literature reviews, as referenced in <a href="#bioengineering-11-01245-t004" class="html-table">Table 4</a>.</p>
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11 pages, 2963 KiB  
Article
Microstructural and Surface Texture Evaluation of Orthodontic Microimplants Covered with Bioactive Layers Enriched with Silver Nanoparticles
by Magdalena Sycińska-Dziarnowska, Magdalena Ziąbka, Katarzyna Cholewa-Kowalska, Gianrico Spagnuolo, Hyo-Sang Park, Steven J. Lindauer and Krzysztof Woźniak
J. Funct. Biomater. 2024, 15(12), 371; https://doi.org/10.3390/jfb15120371 - 9 Dec 2024
Viewed by 103
Abstract
Bacterial infections are a common cause of clinical complications associated with the use of orthodontic microimplants. Biofilm formation on their surfaces and subsequent infection of peri-implant tissues can result in either exfoliation or surgical removal of these medical devices. In order to improve [...] Read more.
Bacterial infections are a common cause of clinical complications associated with the use of orthodontic microimplants. Biofilm formation on their surfaces and subsequent infection of peri-implant tissues can result in either exfoliation or surgical removal of these medical devices. In order to improve the properties of microimplants, hybrid coatings enriched with silver nanoparticles, calcium, and phosphorus were investigated. The present study aimed to assess the microstructure of commercially available microimplants composed of a medical TiAlV (Ti6Al4V) alloy covered with organic–inorganic layers obtained by the sol–gel method using the dip-coating technique. The microstructures and elemental surface compositions of the sterile, etched, and layer-modified microimplants were characterized by scanning electron microscopy with X-ray energy-dispersive spectroscopy (SEM-EDS). Elements such as silver (Ag), calcium (Ca), phosphorus (P), silicon (Si), oxygen (O), and carbon (C) were detected on the microimplant’s surface layer. The SEM observations revealed that control microimplants (unetched) had smooth surfaces with only manufacturing-related embossing, while etching in hydrofluoric acid increased the surface roughness and introduced fluoride onto the microimplants. Layers with only silver nanoparticles reduced the roughness of the implant surface, and no extrusion was observed, while increased roughness and emerging porosity were observed when the layers were enriched with calcium and phosphorus. The highest roughness was observed in the microimplants etched with AgNPs and CaP, while the AgNPs-only layer showed a reduction in the roughness average parameter due to lower porosity. Enhancing the effectiveness of microimplants can be achieved by applying selective surface treatments to different parts. By keeping the outer tissue contact area smooth while making the bone contact area rough to promote stronger integration with bone tissue, the overall performance of the implants can be significantly improved. Full article
(This article belongs to the Special Issue Feature Papers in Dental Biomaterials (2nd Edition))
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<p>SEM images of microimplant surface and EDS analysis of Ti (<b>a</b>–<b>d</b>), Ti-etched (<b>e</b>–<b>h</b>), Ti- etched with layer with AgNPs (<b>i</b>–<b>l</b>), Ti-etched with layer with AgNPs and enriched with CaP (<b>m</b>–<b>p</b>), Ti with layer with AgNPs (<b>r</b>–<b>u</b>), and Ti with layer with AgNPs and enriched with CaP (<b>w</b>–<b>z</b>). Magnification: 130×, 500×, and 1000×.</p>
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<p>SEM images of plate samples composed of Ti6Al4V alloy: layer with AgNPs (<b>a</b>) and layer with AgNPs and CaP (<b>b</b>). Magnification: 500×.</p>
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<p>Confocal laser scanning microscopy images of microimplant surfaces showing the roughness (Ra) of Ti (<b>a</b>), Ti-etched (<b>b</b>), Ti-etched with layer with AgNPs (<b>c</b>), Ti-etched with layer with AgNPs and enriched with CaP (<b>d</b>), Ti with layer with AgNPs (<b>e</b>), and Ti with layer with AgNPs and enriched with CaP (<b>f</b>).</p>
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16 pages, 2198 KiB  
Article
Inhibitory Effects of Gliadin Hydrolysates on BACE1 Expression and APP Processing to Prevent Aβ Aggregation
by Chin-Yu Lin, Cheng-Hong Hsieh, Pei-Yu Lai, Ching-Wei Huang, Yung-Hui Chung, Shang-Ming Huang and Kuo-Chiang Hsu
Int. J. Mol. Sci. 2024, 25(23), 13212; https://doi.org/10.3390/ijms252313212 - 9 Dec 2024
Viewed by 174
Abstract
Alzheimer’s disease (AD), a leading neurodegenerative disorder, is closely associated with the accumulation of amyloid-beta (Aβ) peptides in the brain. The enzyme β-secretase (BACE1), pivotal in Aβ production, represents a promising therapeutic target for AD. While bioactive peptides derived from food protein hydrolysates [...] Read more.
Alzheimer’s disease (AD), a leading neurodegenerative disorder, is closely associated with the accumulation of amyloid-beta (Aβ) peptides in the brain. The enzyme β-secretase (BACE1), pivotal in Aβ production, represents a promising therapeutic target for AD. While bioactive peptides derived from food protein hydrolysates have neuroprotective properties, their inhibitory effects on BACE1 remain largely unexplored. In this study, we evaluated the inhibitory potential of protein hydrolysates from gliadin, whey, and casein proteins prepared using bromelain, papain, and thermolysin. Through in vitro and cellular assays, bromelain-hydrolyzed gliadin (G-Bro) emerged as the most potent BACE1 inhibitor, with an IC50 of 0.408 mg/mL. G-Bro significantly reduced BACE1 expression and amyloid precursor protein (APP) processing in N2a/PS/APP cell cultures, suggesting its potential to attenuate Aβ aggregation. The unique peptide profile of G-Bro likely contributes to its inhibitory effect, with proline residues disrupting β-sheets, lysine residues introducing positive charges that hinder aggregation, hydrophobic residues stabilizing binding interactions, and glutamine residues enhancing solubility and stability. These findings highlight gliadin hydrolysates, particularly G-Bro, as potential natural BACE1 inhibitors with applications in dietary interventions for AD prevention. However, further studies are warranted to elucidate specific peptide interactions and their bioactivity in neural pathways to better understand their therapeutic potential. Full article
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<p>Degree of hydrolysis of proteins hydrolyzed by (<b>a</b>) papain, (<b>b</b>) bromelain, and (<b>c</b>) thermolysin.</p>
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<p>Effect of G-Bro on the viability of N2a/PS/APP cells. Data are expressed as mean ± SD of three independent experiments. Statistical analyses were performed using one-way ANOVA test. Non-treated cells were considered as control. ** <span class="html-italic">p</span> &lt; 0.01 vs. control.</p>
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<p>Effect of G-Bro on BACE1 expression in N2a/PS/APP cells. Data are expressed as mean ± SD of three independent experiments. Statistical analyses were performed using one-way ANOVA test. Band intensity was evaluated with Image J Version 1.54. Non-treated cells were considered as control. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 vs. control.</p>
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<p>Effect of G-Bro on sAPP production in N2a/PS/APP cells. Data are expressed as mean ± SD of three independent experiments. Statistical analyses were performed using one-way ANOVA test. Non-treated cells were considered as control. * <span class="html-italic">p</span> &lt; 0.05 vs. control.</p>
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<p>Effect of G-Bro on Aβ aggregate formation in N2a/PS/APP cells. Data are expressed as mean ± SD of three independent experiments. Statistical analyses were performed using one-way ANOVA test. Non-treated cells were considered as control. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 vs. control.</p>
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<p>The overall framework of the study.</p>
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12 pages, 3478 KiB  
Article
The Effect of Conditioned Medium from Angiopoietin-1 Gene-Modified Mesenchymal Stem Cells on Wound Healing in a Diabetic Mouse Model
by Qiong Deng, Shenzhen Pan, Fangzhou Du, Hongfei Sang, Zhixin Cai, Xiaoyu Xu, Qian Wei, Shuang Yu, Jingzhong Zhang and Chenglong Li
Bioengineering 2024, 11(12), 1244; https://doi.org/10.3390/bioengineering11121244 - 9 Dec 2024
Viewed by 168
Abstract
Introduction: Mesenchymal stem cells (MSCs) have been introduced as a promising treatment for diabetic wounds. The effects of stem cell therapy are thought to be caused by bioactive molecules secreted by stem cells. Stem cell-based gene therapies can target bioactive molecules. Therefore, treatment [...] Read more.
Introduction: Mesenchymal stem cells (MSCs) have been introduced as a promising treatment for diabetic wounds. The effects of stem cell therapy are thought to be caused by bioactive molecules secreted by stem cells. Stem cell-based gene therapies can target bioactive molecules. Therefore, treatment using conditioned medium (CM) derived from genetically engineered stem cells has been proposed as an alternative option for diabetic ulcer care. Methods: MSCs derived from human umbilical cords were obtained and engineered to overexpress the angiogenin-1 gene (MSCsAng1) through plasmid transfection. This study extracted conditioned medium from MSCs (MSC-CM) or MSCsAng1(MSCAng1-CM) for wound treatment applications. Via in vitro experiments, the proangiogenic effects of MSCAng1-CM were assessed via the migration and tube formation of human umbilical vein endothelial cells (HUVECs). Furthermore, the efficacy of MSCAng1-CM in promoting wound healing, re-epithelialization, hair follicle, and angiogenesis was evaluated via a diabetic mouse skin defect model. Results: In vitro assays demonstrated that MSCAng1-CM significantly enhanced HUVECs’ functions, including migration and tube formation. In vivo assays revealed that MSCAng1-CM exhibited notable advancements in healing speed, re-epithelialization, hair follicle, and angiogenesis. Conclusion: These results indicate that MSCAng1-CM can promote wound healing in diabetic mice and make the vascular structure in regenerated tissues more stable without inducing tissue fibrosis, providing a new therapeutic strategy for treating diabetic skin wounds. This provides a valuable theoretical basis for further research on regenerative medicine and cell therapy. Full article
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<p>Construction of MSCs<sup>Ang1</sup> and preparation and characterization of CM. (<b>A</b>) Schematic representation of the lentiviral plasmid carrying Ang1 and GFP linked via a T2A peptide. (<b>B</b>) Immunofluorescence staining of GFP in MSCs<sup>Ang1</sup>. (<b>C</b>) Quantitative analysis of GFP fluorescence intensity in MSCs<sup>Ang1</sup> showed a high GFP-positive expression rate. (<b>D</b>) qPCR analysis revealed that the mRNA expression level of Ang1 in MSCs<sup>Ang1</sup> was significantly elevated compared to MSCs. (<b>E</b>) Ang1 protein levels in MSC-CM and MSC<sup>Ang1</sup>-CM were assessed, demonstrating a significant increase in Ang1 expression in MSC<sup>Ang1</sup>-CM relative to MSC-CM. Total protein content in MSC-CM and MSC<sup>Ang1</sup>-CM was quantified using Coomassie Brilliant Blue staining. Scale bar: 100 μm. All data are presented as mean ± SD. *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of different treatments on HUVECs’ migration and tube formation. (<b>A</b>) Images of HUVECs’ migration changes after 24 h and 48 h of different treatments. (<b>B</b>) Quantitative statistical analysis of cell migration rate showed that compared to Control and MSC-CM, MSC<sup>Ang1</sup>-CM significantly promoted HUVECs’ migration. (<b>C</b>) Number of HUVEC migrations in the transwell chamber after 24 h of different treatments. (<b>D</b>) Quantitative statistical analysis of the number of cell migrations shows that MSC<sup>Ang1</sup>-CM significantly promoted the number of HUVEC migrations compared to Control and MSC-CM. (<b>E</b>) Tubulogenic images of HUVECs after treatment with Control, MSC-CM, and MSC<sup>Ang1</sup>-CM for 24 h. (<b>F</b>) Quantitative statistical analysis of tubulogenesis experiments showed that compared to MSC-CM and Control, the total length of tubes in the MSC<sup>Ang1</sup>-CM group was significantly increased, with statistically significant differences observed among the three groups. Control: Replacement of 50% of the original medium of HUVECs with fresh D/F medium. Scale bar: 100 μm. All data are presented as mean ± SD. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effect of MSC<sup>Ang1</sup>-CM on wound healing in diabetic mice. (<b>A</b>) Dynamic changes and (<b>B</b>) healing rates of diabetic mice wounds under different treatments. (<b>C</b>) Quantitative statistical analysis of dynamic changes in wound healing showed that compared with PBS and MSC-CM groups, the MSC<sup>Ang1</sup>-CM treatment group significantly accelerated the healing of skin wounds in diabetic mice. (<b>D</b>) H&amp;E staining of diabetic mice wound tissues after 14 days of different treatments, orange arrows indicate hair follicles, and green arrows indicate blood vessels. (<b>E</b>) Quantitative statistical analysis of the Epithelial Gap (EG) in diabetic mice wounds after 14 days of treatments with PBS, MSC-CM, and MSC<sup>Ang1</sup>-CM showed that compared with PBS and MSC-CM EGs, the EG in the MSC<sup>Ang1</sup>-CM treatment group was significantly reduced and the re-epithelialization of skin wounds was significantly accelerated. (<b>F</b>) Quantitative statistical analysis of wound thickness in diabetic mice on day 14 showed that the skin thickness in the MSC-CM and MSC<sup>Ang1</sup>-CM groups was increased compared to the PBS group, while there was no significant difference between the MSC-CM and MSC<sup>Ang1</sup>-CM groups. All data are presented as mean ± SD. ns <span class="html-italic">p</span> &gt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>MSC<sup>Ang1</sup>-CM promotes regeneration of hair follicles and formation of blood vessels in diabetic mice wound migration repair area. (<b>A</b>) Dual immunofluorescence staining images of K14 and K17 in diabetic mice wounds after 14 days of different treatments. (<b>B</b>) Quantitative statistical analysis of hair follicle numbers in diabetic mice wounds on day 14 showed that compared with PBS and MSC-CM treatment, the number of K17-positive hair follicles in the migration repair area of the diabetic wound in the MSC<sup>Ang1</sup>-CM treatment group was significantly higher. (<b>C</b>) Dual immunofluorescence staining images of CD34 and αSMA in the wound migration repair area. (<b>D</b>,<b>E</b>) Quantitative statistical analysis of CD34+ vessels and CD34+αSMA+ double-positive mature vessel structures; (<b>D</b>) results showed that, compared to the PBS and MSC-CM groups, the MSC<sup>Ang1</sup>-CM treatment group had a significant increase in CD34+ vascular-like structures; (<b>E</b>) results showed that, compared to the PBS and MSC-CM groups, the MSC<sup>Ang1</sup>-CM treatment group was significantly increased in CD34+α-SMA+ double-positive mature vascular-like structures in diabetic mice wounds. (<b>F</b>) Western blot results showed that, after 14 days of treatment with PBS, MSC-CM, and MSC<sup>Ang1</sup>-CM, the MSC<sup>Ang1</sup>-CM group significantly upregulated the expression level of CD34 in the diabetic skin wound tissues, with β-Actin as the internal reference. (<b>G</b>) Semi-quantitative statistical analysis of CD34 showed that compared to the PBS and MSC-CM groups, the expression level of CD34 in the diabetic skin wound tissues treated with MSC<sup>Ang1</sup>-CM for 14 days was significantly increased, with a statistically significant difference between them. Scale bars: 50 μm, 100 μm. All data are presented as mean ± SD. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Impact of Ang1 secreted by engineered MSCs<sup>Ang1</sup> on fibrosis. (<b>A</b>–<b>C</b>) Expression levels of Col1a1, Col3a1, and the Col1a1/Col3a1 ratio in HUVECs after 24 h of different treatments. The results showed that both Col1a1 and Col3a1 expression levels were elevated in the MSC<sup>Ang1</sup>-CM compared to Control and MSC-CM; however, there were no significant differences in the Col1a1/Col3a1 ratio. (<b>D</b>–<b>F</b>) Expression levels of Col1a1, Col3a1, and the Col1a1/Col3a1 ratio in diabetic skin wound tissues after 14 days of different treatments. The results showed that both Col1a1 and Col3a1 expression levels were elevated in the MSC<sup>Ang1</sup>-CM compared to PBS and MSC-CM; however, there were no significant differences in the Col1a1/Col3a1 ratio among different treatments. Control: Replacement of 50% of the original medium of HUVECs with fresh D/F medium ns <span class="html-italic">p</span> &gt; 0.05, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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11 pages, 3982 KiB  
Communication
Bioactive Agrocomposite for Tissue Engineering and Bone Regeneration
by Miguel Suffo, Celia Pérez-Muñoz, Daniel Goma-Jiménez, Carlos Revenga, Pablo Andrés-Cano and Miguel Ángel Cauqui-López
Inventions 2024, 9(6), 123; https://doi.org/10.3390/inventions9060123 - 9 Dec 2024
Viewed by 177
Abstract
Background: This study describes a novel biomaterial consisting of a mixture of biphasic bioceramic obtained from waste generated by the sugar industry (Carbocal) and a medical-grade epoxy resin adhesive called LOCTITE® M31 CLTM. The objective was to demonstrate the possibility of coating [...] Read more.
Background: This study describes a novel biomaterial consisting of a mixture of biphasic bioceramic obtained from waste generated by the sugar industry (Carbocal) and a medical-grade epoxy resin adhesive called LOCTITE® M31 CLTM. The objective was to demonstrate the possibility of coating non-bioactive and non-biodegradable metallic surfaces on implantable elements. Methods: After preparation, the mixture was applied to the surfaces of hip prostheses composed of two distinct materials: polyetherimide and grade 5 titanium. In both cases, adhesion tests produced favourable results. Additionally, cell cultures were conducted using human foetal osteoblastic cell lines (hFOB 1.19). Results: It was observed that the mixture did not affect the proliferation of bone cells. Conclusions: This composite material was found to promote the growth of bone cells, suggesting its potential for fostering bone tissue development. Full article
(This article belongs to the Section Inventions and Innovation in Biotechnology and Materials)
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<p>Sample preparation: (<b>a</b>) composite materials formed before coating the samples. Nomenclature I–VII corresponds to the compositions indicated in <a href="#inventions-09-00123-t002" class="html-table">Table 2</a>; (<b>b</b>) samples after coating 3 implantable surfaces (PEI-ULTEM1010<sup>®</sup> from the manufacturer Sabic (Riyadh, Saudi Arabia), Vitalium<sup>®</sup> from the manufacturer Dentsply Sirona (Charlotte, NC, USA), and Ti-6Al-4V); (<b>c</b>) implantable surfaces with adhesion test coatings: 1a—on femoral head on Vitalium material; 1b—different concentrations on small samples of Vitalium; 2a—epoxy only on flat surface of the intermediate part of the femoral stem of a hip prosthesis, manufactured in fused deposition (FDM) in material U1010; 2b—mixture VII on flat surface of the intermediate part of the femoral stem of a hip prosthesis, manufactured in FDM in material U1010; 2c—mixture VII on flat surface of the distal part of the femoral stem of a hip prosthesis, on loan from Stryker Iberia S.L. (Alcobendas, Madrid, Spain) in Ti-6Al-4V material.</p>
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<p>Instrument with V-shaped blades at 30° used for the grating in the adhesion test.</p>
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<p>Viability of human osteoblasts after 24 h, 48 h, 72 h, and 7 days of incubation with the different biomaterials tested. Culture medium in the absence of any biomaterial was used as a positive control, while 70% methanol was used as a negative control. X: sample tested; Y: viability (%).</p>
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<p>Growth of cells adhered to the surface of the biomaterial VI; (<b>a</b>) cells fixed with 70% methanol; (<b>b</b>) live cells.</p>
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<p>Growth of cells adhered to the surface of the biomaterial VII; (<b>a</b>) cells fixed with 70% methanol; (<b>b</b>) live cells.</p>
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<p>Viability of human osteoblasts after 24 h, 48 h, 72 h, and 7 days of incubation with samples VI and VII. Culture medium in the absence of any biomaterial was used as a positive control, while 70% methanol was used as a negative control. X: measure of time (hours/days); Y: viability (%).</p>
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<p>Results of adhesion tests of the biomaterial to implantable surfaces; (<b>a</b>) coating applied on the distal part of the Ti5 hip prosthesis; (<b>b</b>) coating applied to the femoral head of the hip prosthesis made of Vitalium; (<b>c</b>) coating applied on the intermediate area of the femoral stem of the hip prosthesis, manufactured in FDM 3D printing in U1010 material. In the enlarged view, two coatings can be seen, type I on the left and type VII on the right.</p>
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9 pages, 1782 KiB  
Proceeding Paper
Advancing Functional Food Innovation: A Patent Landscape Analysis of Lamiaceae Bioactives Through Cooperative Patent Classification Systems
by Reda El Boukhari and Ahmed Fatimi
Biol. Life Sci. Forum 2024, 38(1), 1; https://doi.org/10.3390/blsf2024038001 - 9 Dec 2024
Viewed by 48
Abstract
Medicinal plants from the Lamiaceae family hold significant promise as functional food ingredients due to their high content of essential dietary fiber and bioactive compounds. Lamiaceae plants are rich in phenolic acids, flavonoids, and alkaloids, contributing to their antioxidant and anti-inflammatory properties. This [...] Read more.
Medicinal plants from the Lamiaceae family hold significant promise as functional food ingredients due to their high content of essential dietary fiber and bioactive compounds. Lamiaceae plants are rich in phenolic acids, flavonoids, and alkaloids, contributing to their antioxidant and anti-inflammatory properties. This study utilizes a comprehensive patent analysis to explore recent trends in functional foods developed from Lamiaceae plants. We examined patents from databases using Cooperative Patent Classification (CPC) codes relevant to dietetic products and food compositions. Findings indicate a surge in patents related to Lamiaceae-based dietary supplements, particularly those targeting metabolic health, anti-aging, cognitive function, and bone and liver health. Mentha, Scutellaria, Salvia, and Perilla are the most represented genera, with dietary supplements showing potential in chronic disease prevention. This analysis highlights the growing commercial and therapeutic interest in Lamiaceae-derived functional foods, particularly for preventive health applications. Full article
(This article belongs to the Proceedings of The 4th International Electronic Conference on Nutrients)
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<p>The number of <span class="html-italic">Lamiaceae</span>-based functional food patents granted per year between 2001 and 2024 (up to July 16).</p>
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<p>The top 10 jurisdictions identified by their number of patents related to <span class="html-italic">Lamiaceae</span>-based functional food.</p>
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<p>The main <span class="html-italic">Lamiaceae</span> genera concerned by the collected patents.</p>
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<p>The top 10 plant families associated with <span class="html-italic">Lamiaceae</span> from the collected patent documents.</p>
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<p>A summary of the most cited dietary supplement targets from the studied patent documents.</p>
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10 pages, 826 KiB  
Article
Data Mining for the Characterization of a Paper Prototype Obtained with Bacterial Cellulose Derived from Banana and Pineapple By-Products
by Juan Diego Valenzuela-Cobos, Simón Pérez-Martínez, Manuel Fiallos-Cárdenas and Fabricio Guevara-Viejó
Appl. Sci. 2024, 14(23), 11426; https://doi.org/10.3390/app142311426 - 9 Dec 2024
Viewed by 250
Abstract
The primary objective of this research is to evaluate the feasibility of two of the most prevalent agricultural residues in Ecuador, banana peels and pineapple peels, as a carbon source in the culture medium of Komagataeibacter hansenii for the production of bacterial cellulose [...] Read more.
The primary objective of this research is to evaluate the feasibility of two of the most prevalent agricultural residues in Ecuador, banana peels and pineapple peels, as a carbon source in the culture medium of Komagataeibacter hansenii for the production of bacterial cellulose (BC) and BC-based paper. This analysis includes an assessment of the productivity parameters of the obtained BC and the quality parameters of the BC-based paper, employing multivariate statistical methodologies. The experimental design consisted of fifteen treatments: T1 served as the control using the standard HS medium, while treatments T2–T8 used banana peel extracts (BPE), and treatments T9–T15 used pineapple peel extracts (PPE) at concentrations from 10% to 40% (v/v). Extracts were prepared with tailored pretreatments for each type of peel to optimize bioactive compound recovery. Standardized fermentation and purification conditions were applied, and once the cellulose was obtained, additives and coating agents were incorporated to produce paper samples from each treatment. The results indicated that higher BPE concentrations (T5, T6, T7, and T8) correlated significantly with increased Weight and Yield of BC, as well as improved grammage and water content in the BC-based paper. This highlights that efficient paper production is influenced by the quality of the bacterial cellulose used, with BPE-based media yielding optimal results due to their nutrient composition, which promotes bacterial growth and metabolic activity. This approach suggests a pathway for advancing sustainable and economical paper production. Full article
(This article belongs to the Special Issue Innovative Engineering Technologies for the Agri-Food Sector)
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<p>Biplot for the productivity parameters of BC obtained from the 15 study treatments (T1–T15). The evaluated variables were weight (W), yield (Y), and substrate conversion ratio (SCR).</p>
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<p>Biplot for the quality parameters of BC-based paper obtained from the 15 study treatments (T1–T15). The evaluated variables were grammage (GM), brightness (BG), water content (WC), and opacity (OP).</p>
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21 pages, 1660 KiB  
Article
Impact of Conventional Pasteurization, High Temperature Short Time, Ultra-High Temperature, and Storage Time on Physicochemical Characteristics, Bioactive Compounds, Antioxidant Activity, and Microbiological Quality of Fruit Nectars
by Natalia Polak, Stanisław Kalisz, Elżbieta Hać-Szymańczuk and Bartosz Kruszewski
Foods 2024, 13(23), 3963; https://doi.org/10.3390/foods13233963 - 8 Dec 2024
Viewed by 383
Abstract
Berries are a valuable source of numerous bioactive compounds, and they have an interesting organoleptic profile. Unfortunately, their low storage life determines the need for their preservation. Among the various methods used in this regard, it was decided to use the High Temperature [...] Read more.
Berries are a valuable source of numerous bioactive compounds, and they have an interesting organoleptic profile. Unfortunately, their low storage life determines the need for their preservation. Among the various methods used in this regard, it was decided to use the High Temperature Short Time (HTST) (90 °C/15 s) and Ultra-High Temperature (UHT) (130 °C/5 s) methods to preserve the produced fruit nectar blends (strawberry–blackcurrant and strawberry–chokeberry). For comparison, the nectars were also preserved using conventional pasteurization (90 °C/10 min). Physicochemical, chromatographic, and microbiological determinations were carried out in the tested nectars before and immediately after processing, as well as after 1, 2, 3, 4, and 6 months of refrigerated storage. All methods allowed for the significant inactivation of selected microbial groups. Non-significant changes were observed as a result of HTST and UHT processing in the context of pH, TSS, and titratable acidity. Varied major changes occurred in the content of bioactive components (TPC—decrease or increase by 2–4%, TAC—decrease by 3–20%, vitamin C—decrease by 15–78%), antioxidant activity (decrease or increase by 3–9%), and nephelometric turbidity (decrease or increase by 11–65%). Both nectars showed better quality and nutritional value after the HTST and UHT processes compared to treatment with classic pasteurization. Storage affected the degradation of bioactive compounds, reduced antioxidant activity, increased turbidity, and caused the brightening of samples together with reducing redness and yellowness. Considering the results obtained, it is reasonable to recommend the use of the HTST and UHT methods in industrial conditions for the preservation of liquid fruit and vegetable products such as juices, nectars, and beverages. Full article
(This article belongs to the Special Issue Advancing Food Safety through PCR and Modern Detection Techniques)
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<p>Total color difference (∆E*) between nectar samples. PT—traditional pasteurization, HTST—high temperature short time, UHT—ultra-high temperature, SB—strawberry–blackcurrant nectar, SC—strawberry–chokeberry nectar.</p>
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<p>Heatmap of correlations between content of bioactive compounds (TAC, TPC, vitamin C content, individual anthocyanins), color parameters (L*, a*, b*), and antioxidant activity (AA), based on total results obtained for all samples.</p>
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<p>Chromatograms of anthocyanins present in raw nectars: strawberry–blackcurrant (SB) nectar on the left, and strawberry–chokeberry (SC) nectar on the right. Identified anthocyanins: 1—delphinidin–3–O–glucoside, 2—delphinidin–3–O–rutinoside, 3—cyanidin–3–O–galactoside, 4—cyanidin–3–O–glucoside, 5—cyanidin–3–O–rutinoside, 6—cyanidin–3–O–arabinoside, 7—pelargonidin–3–O–glucoside, 8—pelargonidin–3–O–arabinoside, 9—cyanidin–3–O–xyloside.</p>
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<p>DPPH radical scavenging activity [%] of strawberry–blackcurrant nectar (<b>A</b>) and straw-berry-chokeberry nectar (<b>B</b>); ABTS radical scavenging activity [%] of strawberry–blackcurrant nectar (<b>C</b>) and strawberry–chokeberry nectar (<b>D</b>). HTST—high temperature short time, PT—traditional pasteurization, UHT—ultra-high temperature. Values marked with different capital letters are significantly different (<span class="html-italic">p</span> &lt; 0.05) and concern changes between preservation methods in SB or SC nectars in specific weeks of storage. Values marked with different small letters are significantly different (<span class="html-italic">p</span> &lt; 0.05) and concern changes during storage in SB or SC nectars for specific preservation methods.</p>
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17 pages, 4770 KiB  
Article
Effect of Enterococcus hirae GS22 Fermentation-Assisted Extraction on the Physicochemical and Bioactivities of Sea Cucumber Intestinal Polysaccharides
by Xiqian Tan, Xiaoqing Wang, Fangchao Cui, Ali Zeshan, Dangfeng Wang, Xuepeng Li and Jianrong Li
Molecules 2024, 29(23), 5800; https://doi.org/10.3390/molecules29235800 (registering DOI) - 8 Dec 2024
Viewed by 492
Abstract
The sea cucumber intestine (SI), a secondary product from sea cucumber processing, contains polysaccharides as one of its active ingredients, and fermentation is an effective method for extracting bioactive substances from food by-products. In this study, to explore the effect of Enterococcus hirae [...] Read more.
The sea cucumber intestine (SI), a secondary product from sea cucumber processing, contains polysaccharides as one of its active ingredients, and fermentation is an effective method for extracting bioactive substances from food by-products. In this study, to explore the effect of Enterococcus hirae GS22 fermentation on the extraction of SI polysaccharides, the polysaccharides were extracted through the SI with and without Enterococcus hirae GS22 fermentation, and the obtained polysaccharides were designated as SC-PF and SC-P. The extraction yield, the structural characteristics, and the biological functions of the polysaccharides were then evaluated. The results indicated that Enterococcus hirae GS22 could grow well using SI as the substrate and that fermentation could improve the extraction yield of the polysaccharide from 0.48% to 0.63%, decrease the molecular weight (Mw), and change the monosaccharide composition. The diameter of SC-PF was smaller than SC-P, and the absolute value of the zeta potential of SC-PF was found to be lower than SC-P. Fermentation does not change the functional group or the thermal ability of the polysaccharide. SC-PF had better antioxidant ability than SC-P; the DPPH and superoxide anion scavenging ability were 96.3% and 36.5%, respectively. SC-PF also showed nearly 1.3- and 1.1-fold higher inhibition of α-glucosidase and α-amylase as compared to SC-P. The current results showed that E. hirae GS22 fermentation has the potential to extract SI polysaccharides with better prebiotic abilities. Full article
(This article belongs to the Collection Advances in Glycosciences)
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<p>Bacterial growth and pH variation during fermentation: (<b>a</b>) bacterial count; (<b>b</b>) pH value. Different lowercase letters indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The spectra and morphology analysis results of the polysaccharides: (<b>a</b>) UV spectra; (<b>b</b>) FTIR spectra; (<b>c</b>) SEM image of SC-P (5000×); (<b>d</b>) SEM image of SC-PF (5000×); (<b>e</b>) AFM image.</p>
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<p>Structural and thermal analysis of the polysaccharides: (<b>a</b>) XRD analysis; (<b>b</b>) Congo Red analysis; (<b>c</b>) DSC analysis.</p>
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<p>The antioxidant abilities of the polysaccharides: (<b>a</b>) DPPH scavenging ability; (<b>b</b>) superoxide radical scavenging ability. Different lowercase letters indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Functional properties of the polysaccharides: (<b>a</b>) α-amylase activity inhibition; (<b>b</b>) α-glucosidase activity inhibition; (<b>c</b>) Glc absorption capacity; (<b>d</b>) cholesterol absorption capacity. Different lowercase letters indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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19 pages, 3637 KiB  
Article
Valorization of Hom Thong Banana Peel (Musa sp., AAA Group) as an Anti-Melanogenic Agent Through Inhibition of Pigmentary Genes and Molecular Docking Study
by Pichchapa Linsaenkart, Wipawadee Yooin, Supat Jiranusornkul, Korawan Sringarm, Chaiwat Arjin, Pornchai Rachtanapun, Kittisak Jantanasakulwong, Juan M. Castagnini and Warintorn Ruksiriwanich
Int. J. Mol. Sci. 2024, 25(23), 13202; https://doi.org/10.3390/ijms252313202 - 8 Dec 2024
Viewed by 428
Abstract
Prolonged and unprotected exposure to the environment explicitly influences the development of hyperpigmented lesions. The enzyme tyrosinase (TYR) is a key target for regulating melanin synthesis. Several bioactive compounds derived from plant extracts have been found to possess potent anti-melanogenesis properties against TYR. [...] Read more.
Prolonged and unprotected exposure to the environment explicitly influences the development of hyperpigmented lesions. The enzyme tyrosinase (TYR) is a key target for regulating melanin synthesis. Several bioactive compounds derived from plant extracts have been found to possess potent anti-melanogenesis properties against TYR. In particular, the potential of banana peels from various varieties has garnered interest due to their application in skin hyperpigmentation treatment. A molecular docking study demonstrated interactions between rosmarinic acid, which is predominantly found in all Hom Thong peel extracts, and the active site of TYR (PDB ID: 2Y9X) at residues HIS263, VAL283, SER282, and MET280, with the lowest binding energy of −5.05 kcal/mol, showing the strongest interaction. Additionally, Hom Thong banana peels are rich in phenolic compounds that could inhibit melanin content and tyrosinase activity in both human and mouse melanoma cells. These effects may be attributed to the suppression of gene expression related to melanogenesis, including the regulator gene MITF and pigmentary genes TYR, TRP-1, and DCT, indicating effects comparable to those of the standard treatment groups with arbutin and kojic acid. Our findings indicated the potential of Hom Thong peel extracts as anti-melanogenic agents. Full article
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<p>The molecular interaction profile of mushroom tyrosinase (PDB ID: 2Y9X) and the ligands after molecular docking studies binding poses of (<b>a</b>) L-tyrosine; (<b>b</b>) L-DOPA; (<b>c</b>) β-arbutin; and (<b>d</b>) kojic acid visualized by the BIOVIA Discovery Studio Visualizer.</p>
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<p>The molecular interaction profile of (<b>a</b>) 2D and (<b>b</b>) 3D structures of <span class="html-italic">(R)</span>-rosmarinic acid towards mushroom tyrosinase (PDB ID: 2Y9X) visualized by the BIOVIA Discovery Studio Visualizer and PyMOL, respectively.</p>
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<p>Effects of Hom Thong banana peel extracts on cell viability in (<b>a</b>) G361 human melanoma cells at 48 h; (<b>b</b>) B16 mouse melanoma cells at 48 h; (<b>c</b>) G361 human melanoma cells at 72 h; and (<b>d</b>) B16 mouse melanoma cells at 72 h. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, c, d, e, and f) with <span class="html-italic">p</span> &lt; 0.05. Ct: control, Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>Effects of Hom Thong banana peel extracts on cell viability in (<b>a</b>) G361 human melanoma cells at 48 h; (<b>b</b>) B16 mouse melanoma cells at 48 h; (<b>c</b>) G361 human melanoma cells at 72 h; and (<b>d</b>) B16 mouse melanoma cells at 72 h. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, c, d, e, and f) with <span class="html-italic">p</span> &lt; 0.05. Ct: control, Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>Effects of Hom Thong banana peel extracts and their bioactive compounds on (<b>a</b>) melanin content in G361 human melanoma cells; (<b>b</b>) melanin content in B16 mouse melanoma cells; (<b>c</b>) tyrosinase activity in G361 human melanoma cells; and (<b>d</b>) tyrosinase activity in B16 mouse melanoma cells. Figures of L-dopachrome formation in (<b>e</b>) G361 human melanoma cells and (<b>f</b>) B16 mouse melanoma cells. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, c, and d) with <span class="html-italic">p</span> &lt; 0.05. Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>Effects of Hom Thong banana peel extracts and their bioactive compounds on (<b>a</b>) melanin content in G361 human melanoma cells; (<b>b</b>) melanin content in B16 mouse melanoma cells; (<b>c</b>) tyrosinase activity in G361 human melanoma cells; and (<b>d</b>) tyrosinase activity in B16 mouse melanoma cells. Figures of L-dopachrome formation in (<b>e</b>) G361 human melanoma cells and (<b>f</b>) B16 mouse melanoma cells. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, c, and d) with <span class="html-italic">p</span> &lt; 0.05. Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>Effects of Hom Thong banana peel extracts and their bioactive compounds on relative gene expression of (<b>a</b>) <span class="html-italic">MITF</span> in G361 human melanoma cells; (<b>b</b>) <span class="html-italic">MITF</span> in B16 mouse melanoma cells; (<b>c</b>) <span class="html-italic">TYR</span> in G361 human melanoma cells; (<b>d</b>) <span class="html-italic">TYR</span> in B16 mouse melanoma cells; (<b>e</b>) <span class="html-italic">TRP-1</span> in G361 human melanoma cells; (<b>f</b>) <span class="html-italic">TRP-1</span> in B16 mouse melanoma cells; (<b>g</b>) <span class="html-italic">DCT</span> in G361 human melanoma cells; and (<b>h</b>) <span class="html-italic">DCT</span> in B16 mouse melanoma cells. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, and c) with <span class="html-italic">p</span> &lt; 0.05. Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>Effects of Hom Thong banana peel extracts and their bioactive compounds on relative gene expression of (<b>a</b>) <span class="html-italic">MITF</span> in G361 human melanoma cells; (<b>b</b>) <span class="html-italic">MITF</span> in B16 mouse melanoma cells; (<b>c</b>) <span class="html-italic">TYR</span> in G361 human melanoma cells; (<b>d</b>) <span class="html-italic">TYR</span> in B16 mouse melanoma cells; (<b>e</b>) <span class="html-italic">TRP-1</span> in G361 human melanoma cells; (<b>f</b>) <span class="html-italic">TRP-1</span> in B16 mouse melanoma cells; (<b>g</b>) <span class="html-italic">DCT</span> in G361 human melanoma cells; and (<b>h</b>) <span class="html-italic">DCT</span> in B16 mouse melanoma cells. Data are expressed as the mean ± SD. Significant differences between samples are indicated by different letters (a, b, and c) with <span class="html-italic">p</span> &lt; 0.05. Water: aqueous extract of Hom Thong banana peel, 50EtOH: 50% ethanolic extract of Hom Thong banana peel, 95EtOH: 95% ethanolic extract of Hom Thong banana peel.</p>
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<p>The scheme summarizes melanin biosynthesis.</p>
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20 pages, 883 KiB  
Article
Exploring Hyaluronidase and Alpha-Glucosidase Inhibition Activities of the Hydrothermal Extract of Coffee Silverskin Obtained from a Central Composite Design
by Thavy Kit, Agita Rachmala Ginting, Punnanee Sumpavapol, Lita Chheang and Sudtida Pliankarom Thanasupsin
Processes 2024, 12(12), 2805; https://doi.org/10.3390/pr12122805 - 8 Dec 2024
Viewed by 226
Abstract
Coffee silverskin (CS), the main by-product of coffee roasting production, contains various valuable bioactive compounds in its chemical compositions. Hydrothermal water extraction (HDTE) is one of the promising techniques for valorizing the organic fraction of CS into functional bioactive ingredients, which can be [...] Read more.
Coffee silverskin (CS), the main by-product of coffee roasting production, contains various valuable bioactive compounds in its chemical compositions. Hydrothermal water extraction (HDTE) is one of the promising techniques for valorizing the organic fraction of CS into functional bioactive ingredients, which can be further exploited in various applications. This study aimed to evaluate the hyaluronidase and α-glucosidase inhibition activities of the CS extracts obtained under optimized water extraction conditions. Process optimization was performed using central composite design response surface methodology (CCD-RSM) with a broader range of extraction temperatures (25, 137.5, and 250 °C), reaction times (5, 38.5, and 72 min), and solid-to-liquid ratios (1:10, 1:80, and 1:150). The highest yield of 39.62% was obtained at 137.5 °C, with a reaction time of 72 min and an S/L ratio of 1:80. The total caffeoylquinic acid contents (T-CQA) were quantified based on the sum of three major isomers, including 3-CQA, 4-CQA, and 5-CQA. The results revealed that the highest T-CQA (2.76 ± 0.20 mg/g CS) was significantly obtained (p < 0.05) by subcritical water extraction (SWE) at 143.2 °C with an S/L ratio of 1:10 and an extraction time of 10.41 min. At such conditions, the total phenolic content (TPC), antioxidant properties (AP), and caffeine were 96.13 mg gallic acid equivalence per gram (GAE/g) CS, 20.85 ± 0.17 mg Trolox equivalence per gram (TE/g) CS, and 10.84 ± 1.25 mg/g CS, respectively. The 50% inhibition capacity (IC50) of hyaluronidase and α-glucosidase inhibition of the CS extracted were 5.00 mg/mL and 9.00 mg/mL, respectively. Our results supported the potential direct or indirect applications of CS, such as hydrothermal CS extract (HDT-CSE), in functional food or drinks. Repurposing CS residue to manufacture new products can efficiently reduce the amount of organic waste in landfills, thus conserving resources and energy and contributing to a lower overall carbon footprint in coffee production. Full article
(This article belongs to the Section Environmental and Green Processes)
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<p>Waste management hierarchy for coffee silverskin.</p>
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<p>Schematic diagram of the experimental setup.</p>
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<p>Contour plots for the effects of the extraction temperature (A), reaction time (B), and solid-to-liquid ratio (C): (<b>a</b>–<b>c</b>) on the extraction yield (Y1); (<b>d</b>–<b>f</b>) on the TPC (Y2); (<b>g</b>–<b>i</b>) on the AP (Y3); (<b>j</b>–<b>l</b>) on the T-CQA (Y4).</p>
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<p>HPLC chromatogram of bioactive compounds in the coffee silverskin extract.</p>
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<p>Inhibition activities of 3-CQA, 5-CQA, and CSE.</p>
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<p>Schematic diagram of the experimental setup and key findings.</p>
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15 pages, 3170 KiB  
Article
Preparation and Characterization of Small-Size and Strong Antioxidant Nanocarriers to Enhance the Stability and Bioactivity of Curcumin
by Shanshan Tie, Yujin Yang, Jiawei Ding, Yanyan Li, Mengmeng Xue, Jianrui Sun, Fang Li, Qiuxia Fan, Ying Wu and Shaobin Gu
Foods 2024, 13(23), 3958; https://doi.org/10.3390/foods13233958 - 8 Dec 2024
Viewed by 396
Abstract
The purpose of this study was to design nanocarriers with small-size and antioxidant properties for the effective encapsulation of curcumin. Here, procyanidins, vanillin, and amino acids were used to successfully prepare nanocarriers of a controllable size in the range of 328~953 nm and [...] Read more.
The purpose of this study was to design nanocarriers with small-size and antioxidant properties for the effective encapsulation of curcumin. Here, procyanidins, vanillin, and amino acids were used to successfully prepare nanocarriers of a controllable size in the range of 328~953 nm and to endow antioxidant ability based on a one-step self-assembly method. The reaction involved a Mannich reaction on the phenolic hydroxyl groups of procyanidins, aldehyde groups of vanillin, and amino groups of amino acids. Subsequently, curcumin nanoparticles were prepared by loading curcumin with this nanocarrier, and the encapsulation efficiency of curcumin was 85.97%. Compared with free curcumin, the antioxidant capacity and photothermal stability of the embedded curcumin were significantly improved, and it could be slowly released into simulated digestive fluid. Moreover, using the corticosterone-induced PC12 cell injury model, the cell viability increased by 23.77% after the intervention of curcumin nanoparticles, and the cellular antioxidant capacity was also significantly improved. The nanoparticles prepared in this work can effectively improve the solubility, stability, and bioactivity of curcumin, which provides a reference for the embedding and delivery of other hydrophobic bioactive compounds. Full article
(This article belongs to the Section Food Nutrition)
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<p>Preparation and particle size characterization of nanocarriers. (<b>a</b>) A schematic diagram of the preparation of NCs−1~NCs−8 using PCs, vanillin, and amino acids as raw materials. (<b>b</b>) Particle size, polydispersity index (PDI), and (<b>c</b>) particle size distribution of amino acid-dependent nanocarriers NCs−1~NCs−3. (<b>d</b>) Particle size, PDI, and (<b>e</b>) particle size distribution of PC-dependent nanocarriers NCs−1 and NCs−4~NCs−5. (<b>f</b>) Particle size, PDI, and (<b>g</b>) particle size distribution of vanillin-dependent nanocarriers NCs−1 and NCs−6~NCs−8. Note: the lower letters a, b, and c indicate that there are statistically significant differences between the samples.</p>
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<p>Formation and spectral characterization. (<b>a</b>) Schematic illustration of the reaction pathway of NCs. FTIR spectra of (<b>b</b>) vanillin (Van), Lys, (<b>c</b>) PCs, and NCs. (<b>d</b>) UV-Vis spectra and (<b>e</b>) crystal structure of PCs and NCs.</p>
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<p>Preparation and characterization of Cur NPs. (<b>a</b>) A schematic diagram of preparation, SEM image, and (<b>b</b>) embedding efficiency (EE) of Cur NPs. (<b>c</b>) FTIR spectra, (<b>d</b>) UV−vis spectra and (<b>e</b>) crystal structure of Cur NPs.</p>
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<p>Antioxidant capacity experiment. (<b>a</b>) DPPH and (<b>b</b>) ABTS radical scavenging activities for Cur, NCs, and Cur NPs. Note: the lower letters a, b, and c indicate that there are statistically significant differences between the samples.</p>
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<p>(<b>a</b>) UV irradiation and (<b>b</b>) thermal stability analyses for Cur and Cur NPs. Note: the lower letters a, b, and indicate that there are statistically significant differences between the samples.</p>
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<p>(<b>a</b>) Schematic diagram of simulated digestion and (<b>b</b>) release profile of Cur NPs.</p>
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<p>Cell viability analysis. (<b>a</b>) The effect of different concentrations of CORT on the viability of PC12 cells. Effect of (<b>b</b>) NCs, (<b>c</b>) Cur, and (<b>d</b>) Cur NPs on the cell viability of 400 μM CORT-induced PC12 cells. Note: the lower letters a−e indicate that there are statistically significant differences between the samples.</p>
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<p>Effect of Cur, NCs, and Cur NPs on the levels of (<b>a</b>) T-SOD, (<b>b</b>) CAT, and (<b>c</b>) MDA in PC12 cells induced by CORT. Note: the lower letters a, b, c, and d indicate that there are statistically significant differences between the samples.</p>
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<p>(<b>a</b>) Optical images of Cur NPs and NCs. (<b>b</b>) Optical images and hemolysis rate (HR) of negative control, positive control, Cur NPs, and NCs after treatment of red blood cells.</p>
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15 pages, 1329 KiB  
Article
Antimicrobial Mixture Based on Micronized Kaolinite and Ziziphora Essential Oil as a Promising Formulation for the Management of Infected Wounds
by Aigerim A. Karaubayeva, Tolkyn Bekezhanova, Karlygash Zhaparkulova, Katarzyna Susniak, Jan Sobczynski, Paulina Kazimierczak, Agata Przekora, Krystyna Skalicka-Wozniak, Łukasz Kulinowski, Anna Glowniak-Lipa, Zuryiadda B. Sakipova and Izabela Korona-Głowniak
Int. J. Mol. Sci. 2024, 25(23), 13192; https://doi.org/10.3390/ijms252313192 - 8 Dec 2024
Viewed by 342
Abstract
Kaolinite stands out as a promising natural geomaterial for developing new therapeutic systems aimed at addressing global health challenges, such as multidrug-resistant infections. In this study, we report on the formulation and biological activity of a therapeutic mixture composed of white micronized kaolinite [...] Read more.
Kaolinite stands out as a promising natural geomaterial for developing new therapeutic systems aimed at addressing global health challenges, such as multidrug-resistant infections. In this study, we report on the formulation and biological activity of a therapeutic mixture composed of white micronized kaolinite (KAO) and Ziziphora essential oil (ZEO), intended for topical application on infected wounds. GC–MS analysis revealed that the primary component of ZEO is pulegone, constituting 72.98% of the oil. ZEO demonstrated good bioactivity against bacterial and fungal strains (MIC 1.25–5 mg/mL). Additionally, ZEO at a concentration of 0.0156% (0.156 mg/mL) was found to significantly stimulate collagen synthesis. The antimicrobial activity of the tested KAO–ZEO mixture formulation (30% KAO/0.25% ZEO in an excipient base) showed the highest effectiveness against Candida spp. (MIC 0.08–25 mg/mL) and Gram-positive bacteria (MIC 0.16–25 mg/mL), with lower activity against Gram-negative bacteria (MIC 25–50 mg/mL). Moreover, the KAO–ZEO mixture was nontoxic (cell viability near 100%) to human skin fibroblasts according to the ISO 10993-5 standard and promoted collagen synthesis by skin cells. This is the first documented formulation combining KAO and ZEO, demonstrating significant antimicrobial properties along with the ability to stimulate collagen production in fibroblasts. These properties highlight KAO–ZEO as a promising novel treatment, which may synergize with current care standards and improve wound healing outcomes. Full article
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<p>The GC–MS chromatogram of Ziziphora EO (ZEO).</p>
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<p>The picture of gel formulation containing micronized kaolinite clay and Ziziphora essential oil.</p>
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<p>Evaluation of the biological properties of ZEO and KAO–ZEO on normal human skin fibroblasts (BJ): (<b>a</b>) Screening cytotoxicity test on different concentrations of ZEO ranging from 0.0039% to 0.2500% and cytotoxicity assessment of KAO–ZEO extract prepared by soaking 100 mg of the sample in 1 mL of the culture medium for 24 h at 37 °C; (<b>b</b>) Evaluation of cell proliferation after exposure to the highest non-toxic concentration of ZEO (0.0156%) and KAO–ZEO extract (prepared at the ratio 100 mg/mL); (<b>c</b>) Collagen synthesis assessment after exposure to the highest non-toxic concentration of ZEO (0.0156%) and KAO–ZEO extract (prepared at the ratio 100 mg/mL); (control—cells maintained in the culture medium without the ZEO and KAO–ZEO; * statistically significant results considered at <span class="html-italic">p</span> &lt; 0.05 compared to the control cells according to One-way ANOVA with post hoc Dunnett’s test).</p>
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