RESUMEN

In the pharmaceutical sector, solid lipid nanoparticles (SLN) are vital for drug delivery incorporating a lipid core. Chondroitin sulfate (CHON) is crucial for cartilage health. It is often used in osteoarthritis (OA) treatment. Due to conflicting results from clinical trials on CHON's efficacy in OA treatment, there has been a shift toward exploring effective topical systems utilizing nanotechnology. This study aimed to optimize a solid lipid nanoparticle formulation aiming to enhance CHON permeation for OA therapy. A 3 × 3 × 2 Design of these experiments determined the ideal parameters: a CHON concentration of 0.4 mg/mL, operating at 20,000 rpm speed, and processing for 10 min for SLN production. Transmission electron microscopy analysis confirmed the nanoparticles' spherical morphology, ensuring crucial uniformity for efficient drug delivery. Cell viability assessments showed no significant cytotoxicity within the tested parameters, indicating a safe profile for potential clinical application. The cell internalization assay indicates successful internalization at 1.5 h and 24 h post-treatment. Biopharmaceutical studies supported SLNs, indicating them to be effective CHON carriers through the skin, showcasing improved skin permeation and CHON retention compared to conventional methods. In summary, this study successfully optimized SLN formulation for efficient CHON transport through pig ear skin with no cellular toxicity, highlighting SLNs' potential as promising carriers to enhance CHON delivery in OA treatment and advance nanotechnology-based therapeutic strategies in pharmaceutical formulations.

Asunto(s)

Sulfatos de Condroitina , Nanopartículas , Sulfatos de Condroitina/química , Animales , Porcinos , Nanopartículas/química , Regeneración/efectos de los fármacos , Cartílago/efectos de los fármacos , Cartílago/metabolismo , Osteoartritis/tratamiento farmacológico , Osteoartritis/patología , Supervivencia Celular/efectos de los fármacos , Humanos , Administración Tópica , Nanoestructuras/química , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos/métodos , Piel/efectos de los fármacos , Piel/metabolismo

RESUMEN

3D-printed hydrogel scaffolds biomimicking the extracellular matrix (ECM) are key in cartilage tissue engineering as they can enhance the chondrogenic differentiation of mesenchymal stem cells (MSCs) through the presence of active nanoparticles such as graphene oxide (GO). Here, biomimetic hydrogels were developed by cross-linking alginate, gelatin, and chondroitin sulfate biopolymers in the presence of GO as a bioactive filler, with excellent processability for developing bioactive 3D printed scaffolds and for the bioprinting process. A novel bioink based on our hydrogel with embedded human MSCs presented a cell survival rate near 100% after the 3D bioprinting process. The effects of processing and filler concentration on cell differentiation were further quantitatively evaluated. The nanocomposited hydrogels render high MSC proliferation and viability, exhibiting intrinsic chondroinductive capacity without any exogenous factor when used to print scaffolds or bioprint constructs. The bioactivity depended on the GO concentration, with the best performance at 0.1 mg mL-1. These results were explained by the rational combination of the three biopolymers, with GO nanoparticles having carboxylate and sulfate groups in their structures, therefore, biomimicking the highly negatively charged ECM of cartilage. The bioactivity of this biomaterial and its good processability for 3D printing scaffolds and 3D bioprinting techniques open up a new approach to developing novel biomimetic materials for cartilage repair.

Asunto(s)

Alginatos , Bioimpresión , Diferenciación Celular , Condrogénesis , Sulfatos de Condroitina , Gelatina , Hidrogeles , Células Madre Mesenquimatosas , Nanocompuestos , Impresión Tridimensional , Andamios del Tejido , Humanos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacología , Alginatos/química , Alginatos/farmacología , Gelatina/química , Bioimpresión/métodos , Diferenciación Celular/efectos de los fármacos , Condrogénesis/efectos de los fármacos , Nanocompuestos/química , Andamios del Tejido/química , Hidrogeles/química , Hidrogeles/farmacología , Ingeniería de Tejidos/métodos , Materiales Biomiméticos/química , Materiales Biomiméticos/farmacología , Grafito/química , Grafito/farmacología , Proliferación Celular/efectos de los fármacos , Células Cultivadas

RESUMEN

In this work the DBL3x domain of the erythrocyte membrane protein from Plasmodium Falciparum (PfEMP1), was revisited as a potential molecular target for the development of new drugs against malaria. This protein interacts with chondroitin sulfate A (CSA), a glycosaminoglycan present in the substance fundamental for connective tissues of vertebrates and is implicated in malaria complications in pregnant women. We performed molecular docking and molecular dynamic studies of DBL3x complexed with CSA and five analogues, where the sulfate group was replaced by phosphate, in order to evaluate if the better electrostatic interactions provided by phosphate groups could afford better binders capable of preventing the binding of CSA to DBL3x. Results suggest that all proposed compounds have high affinity towards DBL3x and could bind better to the DBL3x domain of PfEMP1 than CSA, qualifying as potential inhibitors of this protein and, therefore, new potential leads for the drug design against malaria.Communicated by Ramaswamy H. Sarma.

Asunto(s)

Malaria Falciparum , Malaria , Complicaciones Parasitarias del Embarazo , Animales , Antígenos de Protozoos/química , Sulfatos de Condroitina/química , Sulfatos de Condroitina/metabolismo , Sulfatos de Condroitina/farmacología , Eritrocitos/metabolismo , Femenino , Glicosaminoglicanos/metabolismo , Humanos , Malaria/complicaciones , Malaria/metabolismo , Malaria Falciparum/tratamiento farmacológico , Proteínas de la Membrana/metabolismo , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Fosfatos , Placenta/metabolismo , Plasmodium falciparum/química , Embarazo , Complicaciones Parasitarias del Embarazo/metabolismo , Proteínas Protozoarias/química , Sulfatos/metabolismo

RESUMEN

Chitosan/chondroitin sulfate (CHT/CS) curcumin-charged hydrogels were prepared through polyelectrolytic complexation (PEC) following two methodologies (PEC-CUR and PEC-T-CUR) and were applied on apoptosis of HeLa, HT29 and PC3 cancer cells. PEC-T-CUR (ionic liquid (IL) mixed using ultraturrax homogenizer) results show to be far better than for PEC-CUR (IL mixed using magnetic stirring), with IC50 being improved 5.13 times to HeLa cancer cells (from 1675.2 to 326.7 µg mL-1). PECs produced by this methodology presented favorable characteristics, such as particle size, hydrophobicity, pH swelling. Beyond this, the IL was quantitatively recovered in both cases. CUR entrapment levels were hugely loaded into PEC at around 100%. Swelling, dissolution/degradation, and pHpzc assays showed that PECs may positively act in several environments, releasing the CUR, the CHT and CS as well. Characterization through FTIR, SEM, TEM, TGA, DSC, and WAXS confirmed CUR presence in both types of PECs, and cytotoxic studies showed the significant anticancer effects of CUR-containing PECs.

Asunto(s)

Antineoplásicos/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Quitosano/química , Curcumina/química , Portadores de Fármacos/química , Hidrogeles/química , Líquidos Iónicos/química , Nanopartículas/química , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Sulfatos de Condroitina/química , Células HT29 , Células HeLa , Humanos , Hidrogeles/farmacología , Concentración de Iones de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Células PC-3 , Tamaño de la Partícula , Polielectrolitos/química

RESUMEN

OBJECTIVES: To compare the efficacy and safety of a new formulation of a fixed dose combination of glucosamine sulfate (GS; 1500 mg) and bovine chondroitin sulfate (CS; 1200 mg) versus the reference product (RP) in patients with knee osteoarthritis (OA). METHODS: In this multicenter, randomized, single-blind trial, 627 patients with knee osteoarthritis (OA)-Kellgren-Lawrence grades 2 or 3 and mean score ≥ 40 mm in the WOMAC pain subscale-were randomized to receive GS/CS or the RP for 24 weeks. The primary efficacy endpoint was the absolute change in WOMAC pain subscale score. The secondary endpoints included the following: WOMAC total and subscale scores, overall assessment of the disease by the patient and the investigator, SF-12 score, OMERACT-OARSI response rate to the treatment, and rescue medication use. RESULTS: Mean reductions of WOMAC pain score were - 35.1 (sd = 23.2) mm in the GS/CS group and - 36.5 (sd = 24.9) mm in the RP group. The difference between the adjusted means of both treatments confirmed the non-inferiority of GS/CS versus the RP. Improvement was observed in pain, stiffness, physical function and total WOMAC score, as well as in overall OA assessment by the patient and the investigator for both groups. No improvement was observed in SF-12. The rate of OMERACT-OARSI responders was 89.4% in GS/CS group and 87.9% in the RP group. Headache and changes in glucose tolerance were the most frequent treatment-related adverse events. CONCLUSIONS: The new formulation of a fixed-dose combination of glucosamine sulfate and bovine chondroitin sulfate was non-inferior to the RP in symptomatic treatment of knee OA, with a high responder rate and good tolerability profile. TRIAL REGISTRATION: ClinicalTrials.gov; Registration number NCT02830919 ; Date of registration: July 13, 2016; First randomization date: December 05, 2016).

Asunto(s)

Sulfatos de Condroitina/uso terapéutico , Glucosamina/uso terapéutico , Osteoartritis de la Rodilla/tratamiento farmacológico , Brasil , Sulfatos de Condroitina/efectos adversos , Sulfatos de Condroitina/química , Combinación de Medicamentos , Femenino , Glucosamina/efectos adversos , Glucosamina/química , Humanos , Masculino , Persona de Mediana Edad , Método Simple Ciego , Factores de Tiempo

RESUMEN

A benzamidine derivative from diminazene was tested for a novel activity: treatment of primary open-angle glaucoma. This drug was incorporated into mucoadhesive polymeric inserts prepared using chitosan (Chs) and chondroitin sulfate (CS). Of current interest is the mucoadhesion, which increases the contact time with the ocular surface, resulting in improved bioavailability; also, the inserts are made to act as a prolonged release system. In the present work the inserts were prepared by the solvent casting method using different polymeric proportions (30:70, 50:50, 75:25% w/w Chs:CS and 100% Chs). Thermal analysis and infrared spectroscopy both demonstrated physical dispersion of the active drug. The most promising was the 50:50% Chs:CS which demonstrated that it was not fragile and has an in vitro release profile of up to 180 minutes. In addition, it presented greater adhesion strength in relation to the other formulations. These physicochemical results corroborate the in vivo tests performed. In this sense, we also demonstrated that the treatment with the 50:50% insert can control the intraocular pressure (IOP) for at least 3 weeks and prevents damage to the retinal ganglion cells (RGCs) compared to the placebo insert. Thus, this indicates thus that the new drug is quite viable and promising in glaucoma treatment.

Asunto(s)

Agentes Antiglaucoma/administración & dosificación , Agentes Antiglaucoma/química , Preparaciones de Acción Retardada/química , Diminazeno/análogos & derivados , Diminazeno/administración & dosificación , Glaucoma de Ángulo Abierto/tratamiento farmacológico , Animales , Agentes Antiglaucoma/farmacocinética , Agentes Antiglaucoma/uso terapéutico , Quitosano/química , Sulfatos de Condroitina/química , Diminazeno/farmacocinética , Diminazeno/uso terapéutico , Liberación de Fármacos , Glaucoma de Ángulo Abierto/patología , Masculino , Ratas , Ratas Wistar

RESUMEN

Novel poly(vinyl alcohol)/chondroitin sulfate (PVA/CS) composite hydrogels containing hydroxyapatite (HA) or Sr-doped HA (HASr) particles were synthesized by a freeze/thaw method and characterized aiming towards biomedical applications. HA and HASr were synthesized by a wet-precipitation method and added to the composite hydrogels in fractions up to 15 wt%. Physical-chemical characterizations of particles and hydrogels included scanning electron microscopy, energy-dispersive spectroscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, thermogravimetry, porosity, compressive strength/elastic modulus, swelling degree, and cell viability. Particles were irregular in shape and appeared to have narrow size variation. The thermal behavior of composite hydrogels was altered compared to the control (bare) hydrogel. All hydrogels exhibited high porosity. HA/HASr particles reduced total porosity without reducing pore size. The mechanical strength was improved as the fraction of HA or HASr was increased. HASr particles led to a faster water uptake but did not interfere with the total hydrogel swelling capacity. In cell viability essay, increased cell growth (above 120%) was observed in all groups including the control hydrogel, suggesting a bioactive effect. In conclusion, PVA/CS hydrogels containing HA or HASr particles were successfully synthesized and showed promising morphological, mechanical, and swelling properties, which are particularly required for scaffolding.

Asunto(s)

Materiales Biocompatibles/química , Sulfatos de Condroitina/química , Durapatita/química , Alcohol Polivinílico/química , Estroncio/química , Materiales Biocompatibles/síntesis química , Sulfatos de Condroitina/síntesis química , Fuerza Compresiva , Durapatita/síntesis química , Módulo de Elasticidad , Alcohol Polivinílico/síntesis química , Porosidad , Termogravimetría

RESUMEN

This paper describes the synthesis of hybrid hydrogels based on chondroitin sulfate (CS) and mesoporous silica (MCM-41). The combination of the CS network and surface-modified MCM-41 yields resilient hybrids with a high water absorption power and excellent capacity for the removal of methylene blue (MB). In this system, two types of solute transport mechanisms exist: absorption and adsorption. The effect of MCM-41 on the physical-chemical properties of the hydrogels was assessed over a wide pH scale, and the absorption kinetics and isotherms of MB were studied by theoretical models. The mechanical properties of the hydrogels, such as the modulus of elasticity, yield strength, modulus of resilience, and fracture toughness, were significantly improved. This hydrogel exhibited high performance for water absorption (4000 % beyond its initial weight) and removal of MB (3982 ± 123.6 mg g-1), while the pure hydrogel removed 2912 ± 163.8 mg g-1.

Asunto(s)

Sulfatos de Condroitina/química , Hidrogeles/química , Azul de Metileno/aislamiento & purificación , Dióxido de Silicio/química , Contaminantes Químicos del Agua/aislamiento & purificación , Adsorción , Azul de Metileno/análisis , Contaminantes Químicos del Agua/análisis

RESUMEN

Thrombin triggers cellular responses that are crucial for development and progression of cancer, such as proliferation, migration, oncogene expression and angiogenesis. Thus, biomolecules capable of inhibiting this protease have become targets in cancer research. The present work describes the in vitro antitumor properties of a chondroitin sulfate with anti-thrombin activity, isolated from the Litopenaeus vannamei shrimp (sCS). Although the compound was unable to induce cytotoxicity or cell death and/or cell cycle changes after 24 h incubation, it showed a long-term antiproliferative effect, reducing the tumor colony formation of melanoma cells by 75% at 100 µg/mL concentration and inhibiting the anchorage-independent colony formation. sCS reduced 66% of melanoma cell migration in the wound healing assay and 70% in the transwell assay. The compound also decreased melanin and TNF-α content of melanoma cells by 52% and 75% respectively. Anti-angiogenic experiments showed that sCS promoted 100% reduction of tubular structure formation at 100 µg/mL. These results are in accordance with the sCS-mediated in vitro expression of genes related to melanoma development (Cx-43, MAPK, RhoA, PAFR, NFKB1 and VEGFA). These findings bring a new insight to CS molecules in cancer biology that can contribute to ongoing studies for new approaches in designing anti-tumor therapy.

Asunto(s)

Inhibidores de la Angiogénesis , Antineoplásicos , Sulfatos de Condroitina , Melanoma Experimental/tratamiento farmacológico , Penaeidae/química , Inhibidores de la Angiogénesis/química , Inhibidores de la Angiogénesis/aislamiento & purificación , Animales , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Línea Celular Tumoral , Sulfatos de Condroitina/química , Sulfatos de Condroitina/aislamiento & purificación , Sulfatos de Condroitina/farmacología , Melanoma Experimental/metabolismo , Melanoma Experimental/patología , Ratones , Conejos

RESUMEN

Colorectal cancer (CRC) has a high incidence and resistance to conventional treatments. Curcumin (CUR) is a promising natural product in the treatment of CRC with excellent in vitro results. However, its low bioavailability is a limiting factor in clinical applications. To overcome, CUR was incorporated into hydrogels constituted by chitosan (CHT) and chondroitin sulfate (CS), natural biopolymers, capable of controlled release. Hydrogels were synthesized in ionic liquids (ILs, [Hmim][HSO4]) improving the solubility of CHT and the hydrogel properties. Furthermore, CUR was combined with silver nanoparticles (AgNPs) and visible light by Photodynamic Therapy (PDT), which, through the MEO effect (Metal-Enhanced Singlet Oxygen), leads to cell death. It is highlighted the green synthesis of AgNPs using an ultrasound bath. The CHT/CS hydrogels loaded with CUR/AgNPs were properly characterized. Cellular assays showed that the hydrogels (CHT/CS) were not cytotoxic to healthy tissues. However, PDT selective illumination led to inhibition of Caco-2 human colon cancer cells by the CHT/CS/CUR-AgNPs (CC50 = 91.5 µg mL-1 of hydrogel). The cellular uptake assays showed, in addition to the therapeutic action, that the CUR can works as a diagnostic fluorescence probe (theranostic system). Finally, we highlight our commitment to work with reagents, solvents, and methodologies aiming at the principles of green chemistry.

Asunto(s)

Curcumina/química , Hidrogeles/química , Nanopartículas del Metal/química , Polisacáridos/química , Plata/química , Oxígeno Singlete/metabolismo , Apoptosis/efectos de los fármacos , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Quitosano/química , Sulfatos de Condroitina/química , Curcumina/metabolismo , Curcumina/farmacología , Portadores de Fármacos/química , Humanos , Líquidos Iónicos/química , Luz , Nanopartículas del Metal/toxicidad , Solubilidad

RESUMEN

In this study, the materials were synthesized by chemically crosslinking chondroitin sulfate (CS), casein (CAS), and silica nanospheres (SiO2), creating a highly crosslinked network. The hydrogel release profile was adaptable (that is, it could be faster or slower as needed) simply by changing the polymeric proportion. The incorporation of 5% of silica nanospheres, in mass, for all CAS/CS matrices promoted a better-controlled and sustained release of l-dopa, focusing on the matrix based on 70% of CAS, 30% of CS and 5% of silica, whose l-dopa release lasted for 87 h. Besides, hydrogels are cytocompatible. These new hydrogels can be considered highly attractive materials to be used for controlled and sustained drug release purposes, as well as scaffolds and wound dressing systems.

Asunto(s)

Caseínas/química , Sulfatos de Condroitina/química , Preparaciones de Acción Retardada/química , Hidrogeles/química , Nanosferas/química , Dióxido de Silicio/química , Materiales Biocompatibles/química , Reactivos de Enlaces Cruzados/química , Sistemas de Liberación de Medicamentos/métodos , Liberación de Fármacos , Concentración de Iones de Hidrógeno , Polietilenglicoles/química , Polímeros/química

RESUMEN

Surfactants have been used as a tool to improve the properties of polymeric nanoparticles (NPs) and to increase the rate of hydrophobic drug release by means of these nanoparticles. In this context, this study evaluated the effect of lecithin on the characteristics of chitosan (CHI) and chondroitin sulfate (CS) nanoparticles, when applied in curcumin (Curc) release. CHI/CS NPs and CHI/CS/Lecithin NPs were prepared by the ionic gelation method, both as standards and containing curcumin. Simultaneous conductimetric and potentiometric titrations were employed to optimize the interaction between the polymers. NPs with hydrodynamic diameter of ∼130 nm and zeta potential of +60 mV were obtained and characterized by HRTEM; their pore size and surface area were also analyzed by BET method, DLS, FTIR, XPS, and fluorescence spectroscopy techniques to assess morphological and surface properties, stability and interaction between polymers and to quantify the loading of drugs. The final characteristics of NPs were directly influenced by lecithin addition, exhibiting enhanced encapsulation efficiency of curcumin (131.8 µg curcumin per mg CHI/CS/Lecithin/Curc NPs). The release of curcumin occurred gradually through a two-stage process: diffusion-controlled dissolution and release of curcumin controlled by dissolution of the polymer. However, the release of curcumin in buffer solution at pH 7.4 was achieved faster in CHI/CS/Lecithin/Curc NPs than in CHI/CS/Curc NPs. in vitro cytotoxic activity evaluation of the curcumin was determined by the MTT assay, observing that free curcumin and curcumin nanoencapsulated in CHI/CS/Curc and CHI/CS/Lecithin/Curc NPs reduced the viability of MCF-7 cells in the 72 h period (by 28.4, 36.0 and 30.7%, P < 0.0001, respectively). These results indicate that CHI/CS/Lecithin NPs have more appropriate characteristics for encapsulation of curcumin.

Asunto(s)

Quitosano/química , Sulfatos de Condroitina/química , Curcumina/química , Lecitinas/química , Nanopartículas/química , Supervivencia Celular/efectos de los fármacos , Quitosano/administración & dosificación , Sulfatos de Condroitina/administración & dosificación , Curcumina/administración & dosificación , Preparaciones de Acción Retardada/administración & dosificación , Preparaciones de Acción Retardada/química , Liberación de Fármacos , Humanos , Lecitinas/administración & dosificación , Células MCF-7 , Nanopartículas/administración & dosificación

RESUMEN

Nowadays, pharmaceutical heparin is purified from porcine and bovine intestinal mucosa. In the past decade there has been an ongoing concern about the safety of heparin, since in 2008, adverse effects associated with the presence of an oversulfated chondroitin sulfate (OSCS) were observed in preparations of pharmaceutical porcine heparin, which led to the death of patients, causing a global public health crisis. However, it has not been clarified whether OSCS has been added to the purified heparin preparation, or whether it has already been introduced during the production of the raw heparin. Using a combination of different analytical methods, we investigate both crude and final heparin products and we are able to demonstrate that the sulfated contaminants are intentionally introduced in the initial steps of heparin preparation. Furthermore, the results show that the oversulfated compounds are not structurally homogeneous. In addition, we show that these contaminants are able to bind to cells in using well known heparin binding sites. Together, the data highlights the importance of heparin quality control even at the initial stages of its production.

Asunto(s)

Anticoagulantes/aislamiento & purificación , Sulfatos de Condroitina/aislamiento & purificación , Contaminación de Medicamentos , Heparina/aislamiento & purificación , Animales , Anticoagulantes/química , Bovinos , Sulfatos de Condroitina/química , Heparina/química , Liasa de Heparina/química , Humanos , Hidrólisis , Mucosa Intestinal/química , Espectroscopía de Resonancia Magnética , Polisacárido Liasas/química , Control de Calidad , Porcinos

RESUMEN

The parotoid gland of bufonids is characterized as a specialized integument region, formed by different gland types. The secretion elaborated by the largest glandular alveoli has been related to animal chemical defense and is constituted by granular protein content, associated with a basophilic and alcianophilic material with features of glycoconjugates. This study aimed to identify and characterize the glycoconjugates in the secretion of the largest granular gland of the parotoid gland of Rinella icterica by histochemical and immunohistochemical techniques at light microscopy, biochemical methods, and nuclear magnetic resonance spectroscopy. Our results showed that the glycoconjugate content contains a mixture of chondroitin­6­sulfate (C6S) and chondroitin-non-sulfate (C0S). Thus, chondroitin sulfate probably plays an important role in gland physiology, probably protecting the protein content while inside the secretory portion.

Asunto(s)

Acetilgalactosamina/química , Bufonidae/metabolismo , Sulfatos de Condroitina/química , Ácido Glucurónico/química , Glicoconjugados/química , Glándula Parótida/química , Acetilgalactosamina/aislamiento & purificación , Animales , Brasil , Bufonidae/anatomía & histología , Secuencia de Carbohidratos , Sulfatos de Condroitina/aislamiento & purificación , Ácido Glucurónico/aislamiento & purificación , Glicoconjugados/aislamiento & purificación , Masculino , Glándula Parótida/anatomía & histología , Glándula Parótida/fisiología

RESUMEN

High-risk human papillomavirus (HPV) infection, mainly with HPV16 type, has been increasingly considered as an important etiologic factor in head and neck cancers. Detection of HPV16 is therefore crucial for these types of cancer, but clinical tests are not performed routinely in public health systems owing to the high cost and limitations of the existing tests. In this article, we report on a potentially low-cost genosensor capable of detecting low concentrations of HPV16 in buffer samples and distinguishing, with high accuracy, head and neck cancer cell lines according to their HPV16 status. The genosensor consisted of a microfluidic device that had an active layer of a HPV16 capture DNA probe (cpHPV16) deposited onto a layer-by-layer film of chitosan and chondroitin sulfate. Impedance spectroscopy was the principle of detection utilized, leading to a limit of detection of 10.5 pM for complementary ssDNA HPV16 oligos (ssHPV16). The genosensor was also able to distinguish among HPV16+ and HPV16- cell lines, using the multidimensional projection technique interactive document mapping. Hybridization between the ssHPV16 oligos and cpHPV16 probe was confirmed with polarization-modulated infrared reflection-absorption spectroscopy, where PO2 and amide I and amide II bands from adenine and thymine were monitored. The electrical response could be modeled as resulting from an adsorption process represented in a Freundlich model. Because the fabrication procedures of the microfluidic devices and genosensors and the data collection and analysis can be implemented at low cost, the results presented here amount to a demonstration of possible routine screening for HPV infections.

Asunto(s)

Carcinoma de Células Escamosas/virología , Neoplasias de Cabeza y Cuello/virología , Papillomavirus Humano 16/aislamiento & purificación , Técnicas Analíticas Microfluídicas , Infecciones por Papillomavirus/diagnóstico , Adenina/química , Carcinoma de Células Escamosas/diagnóstico , Línea Celular Tumoral , Quitosano/química , Sulfatos de Condroitina/química , ADN de Cadena Simple/química , Impedancia Eléctrica , Neoplasias de Cabeza y Cuello/diagnóstico , Humanos , Límite de Detección , Nanoestructuras/química , Timina/química

RESUMEN

Fucosylated chondroitin sulfates (FCSs) and sulfated fucans (SFs) are conspicuous components of the body wall of sea cucumbers (Holothuroidea). FCSs are composed of a central core of chondroitin sulfate (CS) decorated with branches of mono- or both mono- and disaccharides of α-fucose (FCS types I and II, respectively). FCSs type II have heterogeneous and irregularly distributed α-fucose branches; however, the novel FCS type II from Holothuria lentiginosa described herein via solution nuclear magnetic resonance has strikingly homogeneous α-fucose branches neatly distributed along its CS core. This FCS is built up of three distinct sequential units composed of the typical CS disaccharides of FCSs, rich in ß-galactosamine-4,6diS, decorated with branches of α-Fucp-2,4diS, α-Fucp-3,4diS or α-Fucp[1→3]α-Fucp-4S[1→ linked to the position 3- of the ß-glucuronic acid. Conformational analyses of these repetitive units revealed a fairly rigid structure despite of the high sulfate content of their α-fucose branches. We also determined the structure of the SF from H. lentiginosa as a repetitive tetrasaccharide sequence composed of →3]α-Fucp-2,4diS[1→3]α-Fucp[1→3]α-Fucp-2S[1→3]α-Fucp-2S[1→. Furthermore, we determined that the nonsulfated α-fucose units present in FCS type II did not interfere with their anticoagulant potencies and affinities to calcium. FCS is an autapomorphic molecular character of the class Holothuroidea and the composition of their α-fucose branches differs in a species-specific manner. Branches containing α-Fucp-2,4diS are the most common within the extant holothurians, being found in 90% of the FCSs characterized thus far.

Asunto(s)

Sulfatos de Condroitina/química , Fucosa/química , Holothuria/química , Animales , Conformación de Carbohidratos

RESUMEN

In this study, highly neutralized, highly porous, and ultralight polymeric aerogels prepared from aqueous colloidal suspensions of chitosan (CS) and chondroitin sulfate (ChS) nanocomplexes, formulated as quasi-equimolar amounts of both, are described. These aerogels were designed as healing agents under the inspiration of minimizing the amount of matter applied to wounds, reducing the electrostatic potential of the material and avoiding covalent cross-linkers in order to decrease metabolic stress over wounds. Aerogels synthesized under these criteria are biocompatible and provide specific properties for the induction of wound healing. They do not affect neither the metabolic activity of cultured 3T3 fibroblasts nor the biochemical parameters of experimental animals, open wounds close significantly faster and, unlike control wounds, complete reepithelialization and scarring can be attained 14 days after surgery. Because of its hydration abilities, rapid adaptation to the wound bed and the early accelerator effect of wound closure, the CS/ChS aerogels appear to be functional inducers of the healing. Previous information show that CS/ChS aerogels improve wound bed quality, increase granulation tissue and have pain suppressive effect. CS/ChS aerogels are useful as safe, inexpensive and easy to handle materials for topical applications, such as skin chronic wounds. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2464-2471, 2018.

Asunto(s)

Sulfatos de Condroitina , Ensayo de Materiales , Piel , Cicatrización de Heridas/efectos de los fármacos , Células 3T3 , Administración Tópica , Animales , Quitosano/química , Quitosano/farmacología , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacología , Geles , Masculino , Ratones , Porosidad , Conejos , Piel/lesiones , Piel/metabolismo , Piel/patología

RESUMEN

The occurrence of a natural and unmodified highly sulfated chondroitin sulfate from Litopenaeus vannamei heads (sCS) is herein reported. Its partial digestion by Chondroitinases AC and ABC together with its electrophoretic migration profile revealed it as a highly sulfated chondroitin sulfate despite its average molecular weight being similar to CSA. Using orthogonal 1D/2D NMR experiments, the anomeric signals (δ 4.62/106.0) corresponding to unusual 2,3-di-O-Sulfo-GlcA (∼36%), U33S (δ 4.42/84.1, ∼63%) and U22S (4.12/80.1, ∼50%) substitutions were confirmed. In addition, non-sulfated GlcA (δ 4.5/106.3) linked to 4-O- (A14S, 36%) or 6-O-Sulfo (A16S, 28%) GalNAc (δ 4.64/103.5) was observed. Although the biological role of sCS in shrimp is unknown, its influence on hemostasis was also demonstrated. The sCS identification brings to light new questions about the hierarchical model of GAGs biosynthesis and contributes to the better understanding of the subtle relationship between GAGs structure and function.

Asunto(s)

Anticoagulantes/química , Sulfatos de Condroitina/química , Decápodos/química , Ácido Glucurónico/química , Animales , Anticoagulantes/farmacología , Células Cultivadas , Sulfatos de Condroitina/farmacología , Hemostasis/efectos de los fármacos , Masculino , Conejos , Ratas , Ratas Wistar

RESUMEN

Mesalamine (5-ASA) consists of the first-line therapy for the treatment of ulcerative colitis; however, it has low bioavailability, can cause several systemic adverse events, and has low treatment adherence due to the inconvenient dosing scheme. In this work, a new drug delivery system consisting of chondroitin sulfate linked to 5-ASA was synthesized using a carbodiimide as conjugating agent. The system was characterized by spectroscopic techniques (UV, ATR-FTIR, XRD, and NMR 1H) and thermal analysis (TG/DTG and DSC), suggesting the conjugation between the drug and the polymer. The in vitro release and the corresponding kinetics were also evaluated, revealing that approximately 40% of the drug linked was released at pH9 for up to 50h, following Higuchi's model. The conjugate did not show cytotoxicity for the human monocytic cell line at the doses tested, and an in vivo biodistribution study showed that the conjugate remained in the lower GIT for up to 8h with no uptake in the upper GIT. These data corroborate with the radiation found per segment of GIT and in blood. For this last test the conjugate was radiolabeled with Technetium-99m to allow the scintigraphy evaluation and radiation quantification. In conclusion, the polymeric conjugate was successfully synthesized and demonstrated a mucoadhesiveness on the colon as desired, thus supporting its potential use in the treatment of ulcerative colitis.

Asunto(s)

Sulfatos de Condroitina/química , Portadores de Fármacos/química , Mucosa Intestinal/metabolismo , Mesalamina/administración & dosificación , Profármacos/administración & dosificación , Disponibilidad Biológica , Línea Celular , Colitis Ulcerosa/tratamiento farmacológico , Preparaciones de Acción Retardada , Composición de Medicamentos , Humanos , Mesalamina/farmacocinética , Mesalamina/farmacología , Monocitos/efectos de los fármacos , Monocitos/inmunología , Profármacos/farmacocinética , Profármacos/farmacología , Distribución Tisular , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores

RESUMEN

Fucosylated chondroitin sulfate (FCS), a structurally distinct glycosaminoglycan from the body wall of sea cucumber, possesses many biological properties and pharmacology functions. The refined structure of FCS isolated from sea cucumber Holothuria Mexicana (FCShm) was characterized by NMR spectra and HILIC-FTMS, which demonstrated four types of branches in FCShm. Among these, two branches were α-L-Fuc-2S4S (where Fuc is fucose and S is sulfo) and α-L-Fuc-4S linked to O-3 of glucuronic acid residues, while others were identified as α-L-Fuc-4S and α-L-Fuc-3S4S attached to O-6 of N-acetylgalactosamine residue. Furthermore, the fucosyl branches were α-1,3-linked with different degree of polymerization from 1 to 5. FCShm exhibited high affinity to fibroblast growth factor 1 and 2, growth factors involved in neovascularization. Moreover, FCShm displayed intrinsic anticoagulant activity and inhibited thrombin and factor Xa activation by antithrombin III. Our results proposed a novel structural FCS and demonstrated its favorable application prospects in anti-angiogenesis and anticoagulation.

Asunto(s)

Anticoagulantes/farmacología , Coagulación Sanguínea/efectos de los fármacos , Sulfatos de Condroitina/química , Factores de Crecimiento de Fibroblastos/metabolismo , Holothuria/química , Animales , Bovinos , Cromatografía Líquida de Alta Presión , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Espectroscopía de Resonancia Magnética , Unión Proteica/efectos de los fármacos , Ovinos , Resonancia por Plasmón de Superficie , Sus scrofa