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1.
Tissue Eng Part C Methods ; 28(8): 431-439, 2022 08.
Artículo en Inglés | MEDLINE | ID: mdl-35658609

RESUMEN

Thermal injury trauma can induce a state of immunosuppression, causing wounds to become chronic in nature. Stem cell-based therapies represent a promising new approach to treat such wounds due to their capacity to self-renew and their multi-lineage potential. Mesenchymal stem cells (MSCs) are known to secrete endogenous factors that stimulate wound healing by promoting angiogenesis, extracellular matrix remodeling, skin regeneration, and by dampening down inflammation. MSC delivery in a biomaterial construct can augment their wound-healing capacity by concentrating cells at the burn site and upregulating trophic factor secretion. The work presented is the first to evaluate repair in an in vitro raft thermal injury model using a regenerative, dual cell delivery three-dimensional (3D) core/shell (c/s) "living dressing" construct. This previously characterized 3D c/s bioprinted construct, which delivers both MSCs and endothelial cells, was used to treat an in vitro 3D raft skin thermal injury wound model. The mesenchymal stromal cell line (T0523) was encapsulated within a gelatin-based shell bioink, and human umbilical vein endothelial cells within a chitosan-based core bioink to biofabricate a living dressing for enhanced thermal injury repair and regeneration. We hypothesized that the cell-laden c/s tissue engineered construct (TEC) would strengthen the wound's proangiogenic, anti-inflammatory, and skin regeneration potential. An in vitro thermal injury in a 3D raft skin model showed a slight delay in wound closure in the presence of the c/s TEC but was augmented by corresponding increases in the release of wound-healing factors, epidermal growth factor, matrix metalloproteinases-9, transforming growth factor-α, platelet-derived growth factor; a decrease in pro-inflammatory factor interleukin-6, and evidence of neovascularization.


Asunto(s)
Quemaduras , Cicatrización de Heridas , Vendajes , Quemaduras/terapia , Células Endoteliales , Humanos , Inmunidad
2.
Nutrients ; 14(10)2022 May 19.
Artículo en Inglés | MEDLINE | ID: mdl-35631269

RESUMEN

In the past decades, the regulation of pro-inflammatory cytokine production, including interleukin-8 (IL-8), has been the goal of many targeted therapeutic interventions for Necrotising enterocolitis (NEC), a gastrointestinal disease commonly associated with a very low birth weight in preterm infants. In this study, the ability to regulate the production of IL-8 of the water-soluble non-starch polysaccharide (WS-NSP) from taro corm (Tc-WS-NSP) extracted using a conventional (CE) or improved conventional (ICE) extraction method, of the probiotics Lactobacillus acidophilus, Bifidobacterium breve, and Bifidobacterium infantis, and their synbiotic mixtures were evaluated. The TNF-α stimulated HT-29 cells were incubated with undigested or digested Tc-WS-NSPs (CE or ICE), probiotics, and their synbiotic mixtures with Klebsiella oxytoca, an NEC-positive-associated pathogen. Overall, the synbiotic mixtures of digested Tc-WS-NSP-ICE and high bacterial concentrations of L. acidophilus (5.57 × 109), B. breve (2.7 × 108 CFU/mL), and B. infantis (1.53 × 108) demonstrated higher (42.0%, 45.0%, 43.1%, respectively) ability to downregulate IL-8 compared to the sole use of Tc-WS-NSPs (24.5%), or the probiotics L. acidophilus (32.3%), B. breve (37.8%), or B. infantis (33.1%). The ability demonstrated by the Tc-WS-NSPs, the probiotics, and their synbiotics mixtures to downregulate IL-8 production in the presence of an NEC-positive-associated pathogen may be useful in the development of novel prophylactic agents against NEC.


Asunto(s)
Bifidobacterium breve , Colocasia , Enterocolitis Necrotizante , Probióticos , Simbióticos , Bifidobacterium , Bifidobacterium longum subspecies infantis , Citocinas , Enterocolitis Necrotizante/prevención & control , Humanos , Lactante , Recién Nacido , Recien Nacido Prematuro , Interleucina-8 , Lactobacillus acidophilus , Polisacáridos/farmacología , Probióticos/farmacología , Probióticos/uso terapéutico , Agua
3.
Food Chem ; 368: 130820, 2022 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-34416488

RESUMEN

Two commercially available food grade fungal protease preparations (Fungal Protease 31,000 and Fungal Protease 60000) were found to hydrolyse bovine acid whey proteins but left the beta-lactoglobulin (ß-Lg) intact under the processing conditions used. Comparative analysis before and after hydrolysis of bovine acid whey, by 1D- and 2D-PAGE, RP-HPLC and intact-mass mass spectrometry showed that the ß-Lg remains intact and in high yield after hydrolysis by the fungal proteases. The ß-Lg could be separated from the whey protein peptide hydrolysate by ultrafiltration. Subjecting whey fraction to hydrolysis with the fungal protease preparations provides a procedure, under relatively mild conditions, to generate a highly enriched ß-Lg fraction. ß-Lg is recognised as a valued material in the food, pharmaceutical and cosmetic industries due to its properties such as gelling and foaming. The enriched ß-Lg preparation would also have application in areas such as nanoencapsulation.


Asunto(s)
Lactoglobulinas , Leche , Animales , Bovinos , Hidrólisis , Proteínas de la Leche , Péptido Hidrolasas , Suero Lácteo , Proteína de Suero de Leche
4.
Carbohydr Polym ; 275: 118682, 2022 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-34742412

RESUMEN

Layer-by-layer three-dimensional nanofibrous scaffolds (3DENS) were produced using the electrospinning technique. Interest in using biopolymers and application of electrospinning fabrication techniques to construct nanofibers for biomedical application has led to the development of scaffolds composed of PVA, keratin, and chitosan. To date, PVA/keratin blended nanofibers and PVA/chitosan blended nanofibers have been fabricated and studied for biomedical applications. Electrospun scaffolds comprised of keratin and chitosan have not yet been reported in published literature, thus a novel nanofibrous PVA/keratin/chitosan scaffold was fabricated by electrospinning. The resulting 3DENS were characterized using fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), differential scanning colorimetry (DSC), and thermogravimetric analysis (TGA). Physiochemical properties of the polymer solutions such as viscosity (rheology) and conductivity were also investigated. The 3DENS possess a relatively uniform fibrous structure, suitable porosity, swelling properties, and degradation which are affected by the mass ratio of keratin, and chitosan to PVA. These results demonstrate that PVA/keratin/chitosan 3DENS have the potential for biomedical applications.


Asunto(s)
Quitosano/química , Queratinas/química , Nanofibras/química , Alcohol Polivinílico/química , Andamios del Tejido/química , Animales , Materiales Biocompatibles/química , Biopolímeros/química , Rastreo Diferencial de Calorimetría/métodos , Conductividad Eléctrica , Microscopía Electrónica de Rastreo/métodos , Porosidad , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Termogravimetría/métodos , Ingeniería de Tejidos/métodos , Viscosidad
5.
Carbohydr Polym ; 260: 117768, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712126

RESUMEN

Tissue engineering and regenerative medicine have entered a new stage of development by the recent progress in biology, material sciences, and particularly an emerging additive manufacturing technique, three-dimensional (3D) printing. 3D printing is an advanced biofabrication technique which can generate patient-specific scaffolds with highly complex geometries while hosting cells and bioactive agents to accelerate tissue regeneration. Chitosan hydrogels themselves have been widely used for various biomedical applications due to its abundant availability, structural features and favorable biological properties; however, the 3D printing of chitosan-based hydrogels is still under early exploration. Therefore, 3D printing technologies represent a new avenue to explore the potential application of chitosan as an ink for 3D printing, or as a coating on other 3D printed scaffolds. The combination of chitosan-based hydrogels and 3D printing holds much promise in the development of next generation biomedical implants.


Asunto(s)
Quitosano/química , Hidrogeles/química , Impresión Tridimensional , Materiales Biocompatibles/química , Humanos , Medicina Regenerativa , Ingeniería de Tejidos
6.
Food Chem ; 349: 129210, 2021 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-33582541

RESUMEN

This study investigated the effects of an improved extraction method that utilized freeze-thaw, termed improved conventional extraction (ICE), on the yield and properties of the water-soluble non-starch polysaccharide of taro (Tc-WS-NSP) and compared this method to the conventional extraction (CE) method. The freeze-thaw condition was optimized using response surface methodology (RSM) based on yield. The use of the ICE method resulted in a 227.8% increase in yield of Tc-WS-NSP compared to the CE method. The Tc-WS-NSP-ICE had higher purity, lighter color, larger particle size, and higherζ-potential than Tc-WS-NSP-CE. Both of the samples contain the sugar arabinose, galactose, glucose, and mannose and exhibited comparative FTIR, 1H, and 13C NMR spectra. The Tc-WS-NSP-ICE had a semi-crystalline structure resulting in higher thermal stability and had a higher consistency index than Tc-WS-NSP-CE. Overall, the use of the ICE method provided a simple, efficient, and green alternative to CE for the extraction of Tc-WS-NSP.


Asunto(s)
Fraccionamiento Químico/métodos , Fenómenos Químicos , Colocasia/química , Congelación , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Agua/química , Tamaño de la Partícula , Polisacáridos/análisis , Solubilidad
7.
Bioorg Med Chem ; 29: 115837, 2021 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-33223463

RESUMEN

A series of C-2 derivatized 8-sulfonamidoquinolines were evaluated for their antibacterial activity against the common mastitis causative pathogens Streptococcus uberis, Staphylococcus aureus and Escherichia coli, both in the presence and absence of supplementary zinc (50 µM ZnSO4). The vast majority of compounds tested were demonstrated to be significantly more active against S. uberis when in the presence of supplementary zinc (MICs as low as 0.125 µg/mL were observed in the presence of 50 µM ZnSO4). Compounds 5, 34-36, 39, 58, 79, 82, 94 and 95 were shown to display the greatest antibacterial activity against S. aureus (MIC ≤ 8 µg/mL; both in the presence and absence of supplementary zinc), while compounds 56, 58 and 66 were demonstrated to also exhibit activity against E. coli (MIC ≤ 16 µg/mL; under all conditions). Compounds 56, 58 and 66 were subsequently confirmed to be bactericidal against all three mastitis pathogens studied, with MBCs (≥3log10 CFU/mL reduction) of ≤ 32 µg/mL (in both the presence and absence of 50 µM ZnSO4). To validate the sanitizing activity of compounds 56, 58 and 66, a quantitative suspension disinfection (sanitizer) test was performed. Sanitizing activity (>5log10 CFU/mL reduction in 5 min) was observed against both S. uberis and E. coli at compound concentrations as low as 1 mg/mL (compounds 56, 58 and 66), and against S. aureus at 1 mg/mL (compound 58); thereby validating the potential of compounds 56, 58 and 66 to function as topical sanitizers designed explicitly for use in non-human applications.


Asunto(s)
Amidas/farmacología , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Quinolinas/farmacología , Staphylococcus aureus/efectos de los fármacos , Streptococcus/efectos de los fármacos , Amidas/síntesis química , Amidas/química , Antibacterianos/síntesis química , Antibacterianos/química , Relación Dosis-Respuesta a Droga , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Quinolinas/síntesis química , Quinolinas/química , Relación Estructura-Actividad
8.
Foods ; 9(8)2020 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-32784633

RESUMEN

This study assessed the effect of cow milk (CM) and sheep milk (SM) consumption on the micro-structure, mechanical function, and mineral composition of rat femora in a male weanling rat model. Male weanling rats were fed a basal diet with a 50% reduction in calcium and phosphorus content (low Ca/P-diet) supplemented with either SM or CM. Rats were fed for 28 days, after which the femora were harvested and stored. The femora were analyzed by µ-CT, three-point bending, and inductively coupled plasma-mass spectrometry (ICP-MS). The addition of either milk to the low Ca/P-diet significantly increased (p < 0.05) trabecular bone volume, trabecular bone surface density, trabecular number, cortical bone volume, and maximum force, when compared to rats that consumed only the low Ca/P-diet. The consumption of either milk resulted in a significant decrease (p < 0.05) in trabecular pattern factor, and cortical bone surface to volume ratio when compared to rats that consumed only the low Ca/P-diet. The results were achieved with a lower consumption of SM compared to that of CM (p < 0.05). This work indicates that SM and CM can help overcome the effects on bone of a restriction in calcium and phosphorus intake.

9.
Mar Drugs ; 18(7)2020 Jul 19.
Artículo en Inglés | MEDLINE | ID: mdl-32707634

RESUMEN

Polyhydroxylated naphthoquinones (PHNQs), known as spinochromes that can be extracted from sea urchins, are bioactive compounds reported to have medicinal properties and antioxidant activity. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay showed that pure echinochrome A exhibited a cytotoxic effect on Saos-2 cells in a dose-dependent manner within the test concentration range (15.625-65.5 µg/mL). The PHNQ extract from New Zealand sea urchin Evechinus chloroticus did not induce any cytotoxicity within the same concentration range after 21 days of incubation. Adding calcium chloride (CaCl2) with echinochrome A increased the number of viable cells, but when CaCl2 was added with the PHNQs, cell viability decreased. The effect of PHNQs extracted on mineralized nodule formation in Saos-2 cells was investigated using xylenol orange and von Kossa staining methods. Echinochrome A decreased the mineralized nodule formation significantly (p < 0.05), while nodule formation was not affected in the PHNQ treatment group. A significant (p < 0.05) increase in mineralization was observed in the presence of PHNQs (62.5 µg/mL) supplemented with 1.5 mM CaCl2. In conclusion, the results indicate that PHNQs have the potential to improve the formation of bone mineral phase in vitro, and future research in an animal model is warranted.


Asunto(s)
Conservadores de la Densidad Ósea/farmacología , Huesos/efectos de los fármacos , Calcificación Fisiológica/efectos de los fármacos , Naftoquinonas/farmacología , Osteoblastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Erizos de Mar/química , Animales , Conservadores de la Densidad Ósea/química , Conservadores de la Densidad Ósea/aislamiento & purificación , Conservadores de la Densidad Ósea/toxicidad , Huesos/metabolismo , Huesos/patología , Línea Celular Tumoral , Humanos , Hidroxilación , Naftoquinonas/química , Naftoquinonas/aislamiento & purificación , Naftoquinonas/toxicidad , Osteoblastos/metabolismo , Osteoblastos/patología , Factores de Tiempo
10.
ACS Appl Mater Interfaces ; 12(29): 32328-32339, 2020 Jul 22.
Artículo en Inglés | MEDLINE | ID: mdl-32597164

RESUMEN

Three-dimensional (3D) bioprinting has emerged to create novel cell-based therapies for regenerative medicine applications. Vascularized networks within engineered constructs are required, and toward this end, we report a promising strategy using core-shell (c/s) extrusion 3D-bioprinting technology that employs biomimetic biomaterials to construct regenerative, prevascularized scaffolds for wound care. A custom-designed cell-responsive bioink consisting of a 13% (w/v) cell-laden gelatin methacryloyl (GelMA) shell surrounding a peptide-functionalized, succinylated chitosan (C)/dextran aldehyde (D) cell-laden core was successfully bioprinted resulting in organized microdesigns exhibiting excellent cell viability and subsequent vessel formation. Our templating strategy takes advantage of GelMA's intrinsic thermoreversible properties of low degree of acryloyl functionalization used in combination with a lightly, chemically cross-linked peptide-CD core to serve as temporal structural supports that stabilize during extrusion onto a cooled platform. Mechanical integrity was further strengthened layer-by-layer via GelMA UV photo-cross-linking. We report the first example of GelMA used in combination with a peptide-CD bioink to c/s 3D-bioprint regenerative, prevascularized constructs for wound care. Particular cell adhesion and proteolytic peptide-CD functionalized pair combinations, P15/MMP-2 and P15/cRGD, were found to significantly increase growth of human bone-marrow-derived mesenchymal stems cells (hBMSCs) and human umbilical vein endothelial cells (HUVECs). The constructs delivered two cell types: hBMSCs in the shell bioink and HUVECs within the core bioink. Cord-like, natural microvascularization was shown with endothelial cell marker expression as confirmed by immunofluorescence (IF) staining exhibiting tubelike structures. In addition, in vitro skin wound healing activity of the construct showed a ∼twofold rate of wound closure. Overall, c/s 3D-bioprinted, peptide-CD/GelMA constructs provided the appropriate microenvironment for in vitro stem and endothelial cell viability, delivery, and differentiation. We foresee these custom constructs as representing a fundamental step toward engineering larger scale regenerative, prevascularized tissues.


Asunto(s)
Materiales Biomiméticos/farmacología , Quitosano/farmacología , Dextranos/farmacología , Péptidos/farmacología , Impresión Tridimensional , Cicatrización de Heridas/efectos de los fármacos , Materiales Biomiméticos/química , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Quitosano/química , Dextranos/química , Humanos , Estructura Molecular , Tamaño de la Partícula , Péptidos/química , Propiedades de Superficie
11.
Bioorg Med Chem Lett ; 30(11): 127110, 2020 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-32229060

RESUMEN

A series of substituted sulfonamide bioisosteres of 8-hydroxyquinoline were evaluated for their antibacterial activity against the common mastitis causative pathogens Streptococcus uberis, Staphylococcus aureus and Escherichia coli, both in the presence and absence of supplementary zinc. Compounds 9a-e, 10a-c, 11a-e, 12 and 13 were demonstrated to have MICs of 0.0625 µg/mL against S. uberis in the presence of 50 µM ZnSO4. Against S. aureus compounds 9g (MIC 4 µg/mL) and 11d (MIC 8 µg/mL) showed the greatest activity, whereas all compounds were found to be inactive against E. coli (MIC > 256 µg/mL); again in the presence of 50 µM ZnSO4. All compounds were demonstrated to be significantly less active in the absence of supplementary zinc. Compound 9g was subsequently confirmed to be bactericidal, with an MBC (≥3log10 cfu/mL reduction) of 0.125 µg/mL against S. uberis in the presence of 50 µM ZnSO4. To validate the sanitising activity of compound 9g in the presence of supplementary zinc, a quantitative suspension disinfection (sanitizer) test was performed. In this preliminary test, sanitizing activity (>5log10 reduction of CFU/mL in 5 min) was observed against S. uberis for compound 9g at concentrations as low as 1 mg/mL, validating the potential of this compound to function as a topical sanitizer against the major environmental mastitis-causing microorganism S. uberis.


Asunto(s)
Antibacterianos/química , Oxiquinolina/química , Sulfanilamida/química , Zinc/química , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Interacciones Hidrofóbicas e Hidrofílicas , Pruebas de Sensibilidad Microbiana , Oxiquinolina/farmacología , Staphylococcus aureus/efectos de los fármacos , Streptococcus/efectos de los fármacos , Relación Estructura-Actividad
12.
Mater Sci Eng C Mater Biol Appl ; 110: 110612, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32204061

RESUMEN

Keratins are a family of fibrous proteins anticipated to possess wide-ranging biomedical applications due to their abundance, physicochemical properties and intrinsic biological activity. This review mainly focuses on the biomaterials derived from three major sources of keratins; namely human hair, wool and feather, that have effective applications in tissue engineering, wound healing and drug delivery. This article offers five viewpoints regarding keratin i) an introduction to keratin protein extraction and keratin-based scaffold fabrication methods ii) applications in nerve and bone tissue engineering iii) a review on the keratin dressings applied to different types of wounds to facilitate wound healing and thereby repair the skin iv) the utilization of keratinous materials as a carrier system for therapeutics with a controlled manner v) a discussion regarding the main challenges for using keratin in biomedical applications as well as its future prospects.


Asunto(s)
Queratinas , Ingeniería de Tejidos , Andamios del Tejido/química , Cicatrización de Heridas , Animales , Humanos , Queratinas/química , Queratinas/uso terapéutico
13.
Nutrients ; 12(3)2020 Feb 25.
Artículo en Inglés | MEDLINE | ID: mdl-32106433

RESUMEN

The aim of this study was to investigate the effect of either sheep or cow milk supplementation to a low calcium and phosphorus diet on growth and organ mineral distribution in weanling rats. Rats were fed diets consisting of either a control chow, a 50% reduced calcium and phosphorous chow (low Ca/P), low Ca/P and sheep milk, or low Ca/P and cow milk diet for 28 days. Food intake of the rats, the growth rate of the rats, and the concentrations of minerals in the soft organs and serum were determined. Rats fed the low Ca/P diet alone had lower weight gain than rats consuming either of the milk-supplemented diets (p < 0.05). Both sheep milk and cow milk supplementation overcame the effects of consuming a diet restricted in calcium and phosphorus but the sheep milk was effective at a significantly lower level of milk intake (p < 0.05). Significant differences (p < 0.05) in essential and trace mineral concentrations due to milk type were observed in the kidney, spleen, and liver. For non-essential minerals, significant differences (p < 0.05), related to diet, were observed in all organs for arsenic, cesium, rubidium, and strontium concentrations.


Asunto(s)
Calcio/deficiencia , Enfermedades Carenciales/dietoterapia , Leche , Fosfatos/deficiencia , Oligoelementos/análisis , Animales , Calcio de la Dieta , Bovinos , Enfermedades Carenciales/etiología , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Ratas , Ovinos , Destete
14.
Food Chem ; 316: 126339, 2020 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-32044704

RESUMEN

This study investigated the bioactivities of polyhydroxyl-1,4-naphthoquinone (PHNQ) extracts from Evechinus chloroticus shell waste. PHNQs were extracted from E. chloroticus shells and spines using different solvents and the crude extracts were fractionated by HPLC. The antioxidant activity of the PHNQ extracts were evaluated by the DPPH, ABTS, ORAC and FRAP assays. Ethyl acetate was the best extraction solvent and spine extracts showed better antioxidant activity than shell extracts (p < 0.05). The HPLC fraction containing spinochrome E showed the highest antioxidant activity (p < 0.05). The Minimum Inhibitory Concentration of the PHNQ crude extracts and the HPLC fractions ranged from 250 to 2500 µg/mL depending on the PHNQ extract and microbial species tested. Treatment by PHNQ extracts resulted in alteration of the morphology of the microbial cell wall as observed by transmission electron microscopy. PHNQ extracts also exhibited anti-inflammatory activity in rats (ED50 = 8.26 ± 0.22 µg), comparable to that of Celecoxib (6.12 ± 0.18 µg).


Asunto(s)
Antiinfecciosos/farmacología , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Naftoquinonas/farmacología , Erizos de Mar/química , Animales , Masculino , Pruebas de Sensibilidad Microbiana , Ratas , Ratas Sprague-Dawley
15.
Anaerobe ; 61: 102112, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31629806

RESUMEN

Immuno-modulatory effects of infant gut bacteria were tested on poly(I:C) stimulated HT-29 intestinal epithelial cells. Blautia producta, Bacteroides vulgatus, Bacteroides fragilis and Bacteroides thetaiotaomicron decreased transcription of poly(I:C)-induced inflammatory genes. Modulation of basal level and poly(I:C)-induced IL-8 secretion varied between bacterial species, and between heat treated and non-heat treated bacterial cells.


Asunto(s)
Bacterias , Microbioma Gastrointestinal , Regulación de la Expresión Génica , Transcripción Genética , Células HT29 , Humanos , Lactante , Inflamación/genética , Inflamación/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Mucosa Intestinal/patología
16.
Front Microbiol ; 10: 1995, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31555233

RESUMEN

In this study we investigated the influence of oxygen availability on a phenotypic microtiter screen to identify new, natural product inhibitors of growth for the bovine mastitis-causing microorganisms; Streptococcus uberis, Staphylococcus aureus, and Escherichia coli. Mastitis is a common disease in dairy cattle worldwide and is a major cause of reduced milk yield and antibiotic usage in dairy herds. Prevention of bovine mastitis commonly relies on the application of teat disinfectants that contain either iodine or chlorhexidine. These compounds are used extensively in human clinical settings and increased tolerance to chlorhexidine has been reported in both Gram-positive and Gram-negative microorganisms. As such new, non-human use alternatives are required for the agricultural industry. Our screening was conducted under normoxic (20% oxygen) and hypoxic (<1% oxygen) conditions to mimic the conditions on teat skin and within the mammary gland respectively, against two natural compound libraries. No compounds inhibited E. coli under either oxygen condition. Against the Gram-positive microorganisms, 12 inhibitory compounds were identified under normoxic conditions, and 10 under hypoxic conditions. Data revealed a clear oxygen-dependency amongst compounds inhibiting growth, with only partial overlap between oxygen conditions. The oxygen-dependent inhibitory activity of a naturally occurring quinone, ß-lapachone, against S. uberis was subsequently investigated and we demonstrated that this compound is only active under normoxic conditions with a minimum inhibitory concentration and minimum bactericidal concentration of 32 µM and kills via a reactive oxygen species-dependent mechanism as has been demonstrated in other microorganisms. These results demonstrate the importance of considering oxygen-availability in high-throughput inhibitor discovery.

17.
Biomed Mater ; 14(5): 055013, 2019 08 29.
Artículo en Inglés | MEDLINE | ID: mdl-31318339

RESUMEN

Demand for skin replacements is rapidly increasing as burn and full-thickness wounds are difficult to repair due to the low regeneration capability of innate tissues, as well as the physical drawbacks associated with currently available substitutes. To address this need, an emerging 3D printing technique, melt-electrowriting (MEW) was used to create novel bioactive scaffolds to promote skin regeneration. Polycaprolactone (PCL), a bioresorbable and biocompatible, synthetic polymer with Food and Drug Administration approval for use in the human body was selected as scaffold material due to its mechanical stability, flexibility, and superior melt processing properties. In order to increase PCL's biological functionality bioactive milk proteins (MPs) were blended with PCL. To date, this is the first study of its kind detailing the tissue regenerative capacity of PCL containing MPs as bioactive additives for skin regeneration using MEW. The aim of this study was to MEW MP/PCL tissue engineered constructs (TEC) and assess their suitability for generating tissue in vitro. The MPs, lactoferrin (LF) and whey protein (WP), were mixed with PCL individually at varying concentrations (0.05%, 0.1%, 0.25%), and in combination (COMB) at concentrations of 0.25% each. TECs were characterised chemically, physically, and their biological activity assessed in vitro. Physical characterisation of MEW MP/PCL scaffolds showed that reproducible, layered micron range scaffolds could be fabricated; displaying high porosity, low degradation, and rapid protein release. Biological activity, determined via an in vitro skin model using human keratinocytes (HaCaTs) and normal human dermal fibroblasts cells, showed significantly increased cell growth, spreading, and infiltration into LF (0.25%) containing scaffolds and COMB scaffolds when compared to PCL alone (p ≤ 0.05). These findings demonstrated that the combined addition of LF and WP increased the biological activity of MEW PCL scaffolds and could be potentially used as a TEC for deep tissue dermal regeneration.


Asunto(s)
Materiales Biocompatibles/química , Proteínas de la Leche/química , Poliésteres/química , Impresión Tridimensional , Regeneración , Piel/patología , Animales , Bovinos , Línea Celular , Supervivencia Celular , Electroquímica , Fibroblastos/metabolismo , Humanos , Queratinocitos/metabolismo , Lactoferrina/química , Polímeros/química , Porosidad , Piel/metabolismo , Porcinos , Temperatura , Ingeniería de Tejidos/métodos , Andamios del Tejido , Cicatrización de Heridas
18.
Immun Ageing ; 16: 15, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31333751

RESUMEN

BACKGROUND: Cytokines, chemokines, C-reactive proteins (CRP) and ferritin are known inflammatory markers. However, cytokines such as interleukin (IL-1ß), (IL-6) and tumour necrosis factor (TNF-α) have been reported to interfere with both the bone resorption and bone formation processes. Similarly, immune cell cytokines are known to contribute to inflammation of the adipose tissue especially with obesity. IL-10 but not IL-33 has been linked to lower ferritin levels and anemia. In this study, we hypothesized that specific cytokine levels in the plasma of women with low bone mineral density (BMD) would be higher than those in the plasma of healthy women due to the actions of elevated levels of pro-inflammatory cytokines in inducing osteoclast formation and differentiation during senescence. RESULTS: Levels of cytokines (IFNα2, IFN-γ, IL-12p70, IL-33) and monocyte chemoattractant protein-1 (MCP-1) were significantly higher in the plasma of the osteoporotic group compared to the osteopenic and/or healthy groups. Meanwhile CRP levels were significantly lower in women with osteoporosis (P = 0.040) than the osteopenic and healthy groups. Hip BMD values were significantly lower in women with high/detectable values of IL-1ß (P = 0.020) and IL-6 (P = 0.030) compared to women where these were not detected. Similarly, women with high/detectable values of IL-1ß had significantly lower spine BMD than those where IL-1ß was not detected (P = 0.030). Participants' CRP levels were significantly positively correlated with BMI, fat mass and fat percentage (P < 0.001). In addition, ferritin levels of women with high/detectable values of anti-osteoclastogenic IL-10 (P = 0.012) and IL-33 (P = 0.017) were significantly lower than those where these were not detected. There was no statistically significant association between TNF-α and BMD of the hip and lumbar spine. CONCLUSIONS: High levels of cytokines (IFNα2, IFN-γ, IL-12p70, IL-33) and MCP-1 in apparently healthy postmenopausal women are associated with bone health issues. In addition, an increase in levels of IL-10 and IL-33 may be associated with low ferritin levels in this age group. TRIAL REGISTRATION: ANZCTR, ACTRN12617000802303. Registered May 31st, 2017, https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=373020.

19.
Adv Food Nutr Res ; 87: 187-254, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30678815

RESUMEN

Consumer demand for convenience has led to large quantities of seafood being value-added processed before marketing, resulting in large amounts of marine by-products being generated by processing industries. Several bioconversion processes have been proposed to transform some of these by-products. In addition to their relatively low value conventional use as animal feed and fertilizers, several investigations have been reported that have demonstrated the potential to add value to viscera, heads, skins, fins, trimmings, and crab and shrimp shells by extraction of lipids, bioactive peptides, enzymes, and other functional proteins and chitin that can be used in food and pharmaceutical applications. This chapter is focused on reviewing the opportunities for utilization of these marine by-products. The chapter discusses the various products and bioactive compounds that can be obtained from seafood waste and describes various methods that can be used to produce these products with the aim of highlighting opportunities to add value to these marine waste streams.


Asunto(s)
Organismos Acuáticos , Suplementos Dietéticos , Residuos Industriales , Animales , Manipulación de Alimentos , Industria de Alimentos , Humanos
20.
Food Chem ; 272: 379-387, 2019 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-30309558

RESUMEN

The extraction of polyhydroxylated naphthoquinone (PHNQ) pigments from the shell and spines of the New Zealand sea urchin Evechinus chloroticus was evaluated using six different macroporous resins as an alternative to using organic solvent extraction alone. Four of the resins evaluated in this study (D4006, D4020, D101 and NKA-9) provided the best extraction of PHNQ pigments in terms of the overall adsorption and desorption of E. chloroticus PHNQ pigments from the resins. Organic solvents alone had a higher yield of PHNQs than the resins. The PHNQ composition was characterised by high-performance liquid chromatography (HPLC) with diode-array detection and mass spectrometry. Five PHNQ compounds (spinochromes E, B, C, A and echinochrome A), and three aminated PHNQ compounds (spinamine E, echinamines A and B) were identified. The pigments were found to be prone to degradation on exposure to light, with the aminated PHNQ pigments being the least stable.


Asunto(s)
Exoesqueleto/química , Naftoquinonas/química , Naftoquinonas/aislamiento & purificación , Erizos de Mar/química , Columna Vertebral/química , Animales , Estabilidad de Medicamentos , Hidroxilación , Espectrometría de Masas
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