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1.
J Cardiovasc Electrophysiol ; 31(12): 3293-3301, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32966655

RESUMEN

INTRODUCTION: Therapies for substrate-related arrhythmias include ablation or drugs targeted at altering conductive properties or disruption of slow zones in heterogeneous myocardium. Conductive compounds such as carbon nanotubes may provide a novel personalizable therapy for arrhythmia treatment by allowing tissue homogenization. METHODS: A nanocellulose carbon nanotube-conductive hydrogel was developed to have conduction properties similar to normal myocardium. Ex vivo perfused canine hearts were studied. Electroanatomic activation mapping of the epicardial surface was performed at baseline, after radiofrequency ablation, and after uniform needle injections of the conductive hydrogel through the injured tissue. Gross histology was used to assess distribution of conductive hydrogel in the tissue. RESULTS: The conductive hydrogel viscosity was optimized to decrease with increasing shear rate to allow expression through a syringe. The direct current conductivity under aqueous conduction was 4.3 × 10-1 S/cm. In four canine hearts, when compared with the homogeneous baseline conduction, isochronal maps demonstrated sequential myocardial activation with a shift in direction of activation to surround the edges of the ablated region. After injection of the conductive hydrogel, isochrones demonstrated conduction through the ablated tissue with activation restored through the ablated tissue. Gross specimen examination demonstrated retention of the hydrogel within the tissue. CONCLUSIONS: This proof-of-concept study demonstrates that conductive hydrogel can be injected into acutely disrupted myocardium to restore conduction. Future experiments should focus on evaluating long-term retention and biocompatibility of the hydrogel through in vivo experimentation.


Asunto(s)
Hidrogeles , Nanotubos de Carbono , Animales , Perros , Conductividad Eléctrica , Frecuencia Cardíaca , Miocardio
2.
Phys Chem Chem Phys ; 22(26): 14579-14590, 2020 Jul 08.
Artículo en Inglés | MEDLINE | ID: mdl-32597442

RESUMEN

The formation of strongly elastic physical gels based on poly(alkylene oxide)-grafted hyaluronan or carboxymethylcellulose, exhibiting both shear-thickening and strain-hardening have been studied using rheometry and explained using a slightly different interpretation of the transient network theory. The graft copolymers were prepared by a quantitative coupling reaction. Their aqueous solutions displayed a thermoreversible continuous transition from Newtonian fluid to viscoelastic solid which could be controlled by the reaction conditions. The evolution of all material properties of the gel could be categorized into two distinct temperature regimes with a fast evolution at low temperatures followed by a slow evolution at high temperatures. The activation energy of the zero shear viscosity and the relaxation time of the graft inside the interconnecting microdomains were almost identical to each other in both temperature regimes. This suggests that the number of microdomains remained approximately constant whereas the aggregation number inside the microdomains increased according to the binodal curve of the thermosensitive graft.


Asunto(s)
Carboximetilcelulosa de Sodio/química , Ácido Hialurónico/química , Hidrogeles/química , Polímeros/química , Frío , Calor , Transición de Fase , Reología , Sustancias Viscoelásticas/química , Viscosidad
3.
Biomacromolecules ; 20(3): 1271-1280, 2019 03 11.
Artículo en Inglés | MEDLINE | ID: mdl-30681838

RESUMEN

A series of thermoresponsive graft copolymers, gelling at physiological conditions in aqueous solution and cell growth media, have been synthesized using quantitative coupling between a small set of amino-functionalized poly(alkylene oxide) copolymers (PAO) and the carboxylate of the biologically important polysaccharides (PSa) carboxymethylcellulose and the less reactive hyaluronate. Quantitative grafting enables the establishment of structure-function relationship which is imperative for controlling the properties of in situ gelling hydrogels. The EDC/NHS-mediated reaction was monitored using SEC-MALLS, which revealed that all PAOs were grafted onto the PSa backbone. Aqueous solutions of the graft copolymers were Newtonian fluids at room temperatures and formed reversible physical gels at elevated temperatures which were noncytotoxic toward chondrocytes. The established structure-function relationship was most clearly demonstrated by inspecting the thermogelling strength and the onset of thermogelling in a phase diagram. The onset of the thermogelling function could be controlled by the global PAO concentration, independent of graft ratio.


Asunto(s)
Carboximetilcelulosa de Sodio/química , Ácido Hialurónico/química , Polímeros/química , Alquenos/química , Cromatografía en Gel , Óxidos/química , Relación Estructura-Actividad , Termodinámica
4.
Biomacromolecules ; 20(3): 1263-1270, 2019 03 11.
Artículo en Inglés | MEDLINE | ID: mdl-30689362

RESUMEN

The molecular solubility of softwood arabinoglucuronoxylan (AGX) has been thoroughly investigated, and it has been shown that the chemical and physical structures of the extracted hemicellulose are not significantly influenced by different purification steps, but a transient molecular solubility of AGX was observed in aqueous media at low concentrations (1 g/L) when the dissolved macromolecules had a hydrodynamic diameter of up to 10 nm. A phase separation was detected when the concentration was increased to 15 g/L leading to an association of the smaller molecules into fractal structures with a considerably larger diameter, even though the dispersions were still transparent to ocular inspection. Dynamic Light Scattering and Cryo-Transmission Electron Microscopy showed dimensions in the range of 1000 nm. The phase separation of the sample was further characterized by estimating the χ-interaction parameter of AGX in water using the Flory-Huggins theory, and the results supported that water is a poor solvent for AGX. This behavior is crucial when films and hydrogels based on these biopolymers are made, since the association will dramatically affect barrier and mechanical properties of films made from these materials.


Asunto(s)
Picea/química , Xilanos/química , Biopolímeros/química , Microscopía por Crioelectrón , Dispersión Dinámica de Luz , Solubilidad , Xilanos/aislamiento & purificación
5.
Plant Physiol ; 175(4): 1579-1592, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29070516

RESUMEN

Xylan is tightly associated with cellulose and lignin in secondary plant cell walls, contributing to its rigidity and structural integrity in vascular plants. However, the molecular features and the nanoscale forces that control the interactions among cellulose microfibrils, hemicelluloses, and lignin are still not well understood. Here, we combine comprehensive mass spectrometric glycan sequencing and molecular dynamics simulations to elucidate the substitution pattern in softwood xylans and to investigate the effect of distinct intramolecular motifs on xylan conformation and on the interaction with cellulose surfaces in Norway spruce (Picea abies). We confirm the presence of motifs with evenly spaced glycosyl decorations on the xylan backbone, together with minor motifs with consecutive glucuronation. These domains are differently enriched in xylan fractions extracted by alkali and subcritical water, which indicates their preferential positioning in the secondary plant cell wall ultrastructure. The flexibility of the 3-fold screw conformation of xylan in solution is enhanced by the presence of arabinofuranosyl decorations. Additionally, molecular dynamic simulations suggest that the glycosyl substitutions in xylan are not only sterically tolerated by the cellulose surfaces but that they increase the affinity for cellulose and favor the stabilization of the 2-fold screw conformation. This effect is more significant for the hydrophobic surface compared with the hydrophilic ones, which demonstrates the importance of nonpolar driving forces on the structural integrity of secondary plant cell walls. These novel molecular insights contribute to an improved understanding of the supramolecular architecture of plant secondary cell walls and have fundamental implications for overcoming lignocellulose recalcitrance and for the design of advanced wood-based materials.


Asunto(s)
Celulosa/química , Picea/química , Xilanos/química , Conformación de Carbohidratos , Madera/química , Madera/citología
6.
Angew Chem Int Ed Engl ; 57(37): 11987-11992, 2018 09 10.
Artículo en Inglés | MEDLINE | ID: mdl-30044516

RESUMEN

The heterogeneous nature of non-cellulosic polysaccharides, such as arabinoxylan, makes it difficult to correlate molecular structure with macroscopic properties. To study the impact of specific structural features of the polysaccharides on crystallinity or affinity to other cell wall components, collections of polysaccharides with defined repeating units are required. Herein, a chemoenzymatic approach to artificial arabinoxylan polysaccharides with systematically altered branching patterns is described. The polysaccharides were obtained by glycosynthase-catalyzed polymerization of glycosyl fluorides derived from arabinoxylan oligosaccharides. X-ray diffraction and adsorption experiments on cellulosic surfaces revealed that the physicochemical properties of the synthetic polysaccharides strongly depend on the specific nature of their substitution patterns. The artificial polysaccharides allow structure-property relationship studies that are not accessible by other means.

7.
J Biol Chem ; 291(27): 14120-14133, 2016 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-27129264

RESUMEN

Glucuronic acid (GlcAp) and/or methylglucuronic acid (MeGlcAp) decorate the major forms of xylan in hardwood and coniferous softwoods as well as many cereal grains. Accordingly, the complete utilization of glucuronoxylans or conversion to sugar precursors requires the action of main chain xylanases as well as α-glucuronidases that release the α- (1→2)-linked (Me)GlcAp side groups. Herein, a family GH115 enzymefrom the marine bacterium Saccharophagus degradans 2-40(T), SdeAgu115A, demonstrated activity toward glucuronoxylan and oligomers thereof with preference toward MeGlcAp linked to internal xylopyranosyl residues. Unique biochemical characteristics of NaCl activation were also observed. The crystal structure of SdeAgu115A revealed a five-domain architecture, with an additional insertion C(+) domain that had significant impact on the domain arrangement of SdeAgu115A monomer and its dimerization. The participation of domain C(+) in substrate binding was supported by reduced substrate inhibition upon introducing W773A, W689A, and F696A substitutions within this domain. In addition to Asp-335, the catalytic essentiality of Glu-216 was revealed by site-specific mutagenesis. A primary sequence analysis suggested that the SdeAgu115A architecture is shared by more than half of GH115 members, thus defining a distinct archetype for GH115 enzymes.


Asunto(s)
Gammaproteobacteria/enzimología , Glicósido Hidrolasas/metabolismo , Secuencia de Aminoácidos , Dicroismo Circular , Glicósido Hidrolasas/química , Biología Marina , Modelos Moleculares , Conformación Proteica , Homología de Secuencia de Aminoácido
8.
Biomacromolecules ; 16(5): 1489-96, 2015 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-25806996

RESUMEN

The introduction of 3D bioprinting is expected to revolutionize the field of tissue engineering and regenerative medicine. The 3D bioprinter is able to dispense materials while moving in X, Y, and Z directions, which enables the engineering of complex structures from the bottom up. In this study, a bioink that combines the outstanding shear thinning properties of nanofibrillated cellulose (NFC) with the fast cross-linking ability of alginate was formulated for the 3D bioprinting of living soft tissue with cells. Printability was evaluated with concern to printer parameters and shape fidelity. The shear thinning behavior of the tested bioinks enabled printing of both 2D gridlike structures as well as 3D constructs. Furthermore, anatomically shaped cartilage structures, such as a human ear and sheep meniscus, were 3D printed using MRI and CT images as blueprints. Human chondrocytes bioprinted in the noncytotoxic, nanocellulose-based bioink exhibited a cell viability of 73% and 86% after 1 and 7 days of 3D culture, respectively. On the basis of these results, we can conclude that the nanocellulose-based bioink is a suitable hydrogel for 3D bioprinting with living cells. This study demonstrates the potential use of nanocellulose for 3D bioprinting of living tissues and organs.


Asunto(s)
Bioimpresión , Cartílago/crecimiento & desarrollo , Impresión Tridimensional , Ingeniería de Tejidos , Alginatos/química , Animales , Cartílago/química , Celulosa/química , Celulosa/metabolismo , Condrocitos/química , Condrocitos/citología , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Medicina Regenerativa , Andamios del Tejido/química
9.
Appl Microbiol Biotechnol ; 99(11): 4785-94, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25690311

RESUMEN

This work investigates the feasibility of the use of irreversible electroporation (IRE) in the biofabrication of 3D cellulose nanofibril networks via the bacterial strain Gluconacetobacter xylinus. IRE uses electrical pulses to increase membrane permeability by altering the transmembrane potential; past a threshold, damage to the cell becomes too great and leads to cell death. We hypothesized that using IRE to kill the bacteria at specific locations and particular times, we could introduce conduits in the overall scaffold by preventing cellulose biosynthesis locally. Through mathematical modeling and experimental techniques, electrical effects were investigated and the parameters for IRE of G. xylinus were determined. We found that for a specific set of parameters, an applied electric field of 8 to 12.5 kV/cm, producing a local field of 3 kV/cm, was sufficient to kill most of the bacteria and create a localized pore. However, an applied electric field of 17.5 kV/cm was required to kill all. Results suggest that IRE may be an effective tool to create scaffolds with appropriate porosity for orthopedic applications. Ideally, these engineered scaffolds could be used to successfully treat osteochondral defects.


Asunto(s)
Celulosa/química , Celulosa/metabolismo , Electroporación , Gluconacetobacter/metabolismo , Nanofibras/química , Ingeniería de Tejidos/métodos , Andamios del Tejido , Viabilidad Microbiana
10.
Biomacromolecules ; 15(5): 1727-36, 2014 May 12.
Artículo en Inglés | MEDLINE | ID: mdl-24678830

RESUMEN

(1,3)(1,4)-ß-D-Glucan (mixed-linkage glucan or MLG), a characteristic hemicellulose in primary cell walls of grasses, was investigated to determine both its role in cell walls and its interaction with cellulose and other cell wall polysaccharides in vitro. Binding isotherms showed that MLG adsorption onto microcrystalline cellulose is slow, irreversible, and temperature-dependent. Measurements using quartz crystal microbalance with dissipation monitoring showed that MLG adsorbed irreversibly onto amorphous regenerated cellulose, forming a thick hydrogel. Oligosaccharide profiling using endo-(1,3)(1,4)-ß-glucanase indicated that there was no difference in the frequency and distribution of (1,3) and (1,4) links in bound and unbound MLG. The binding of MLG to cellulose was reduced if the cellulose samples were first treated with certain cell wall polysaccharides, such as xyloglucan and glucuronoarabinoxylan. The tethering function of MLG in cell walls was tested by applying endo-(1,3)(1,4)-ß-glucanase to wall samples in a constant force extensometer. Cell wall extension was not induced, which indicates that enzyme-accessible MLG does not tether cellulose fibrils into a load-bearing network.


Asunto(s)
Pared Celular/química , Celulosa/química , Glucanos/química , Glucanos/metabolismo , Triticum/química , Zea mays/química , Adsorción , Pared Celular/metabolismo , Celulosa/metabolismo , Hidrogeles/síntesis química , Hidrogeles/química , Tamaño de la Partícula , Tecnicas de Microbalanza del Cristal de Cuarzo , Propiedades de Superficie , Triticum/citología , Zea mays/citología
11.
Biomacromolecules ; 15(3): 924-30, 2014 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-24495173

RESUMEN

This study focused on the assembly characteristics of debranched xylan onto cellulose surfaces. A rye arabinoxylan polymer with an initial arabinose/xylose ratio of 0.53 was debranched with an oxalic acid treatment as a function of time. The resulting samples contained reduced arabinose/xylose ratios significantly affecting the molecular architecture and solution behavior of the biopolymer. With this treatment, an almost linear xylan with arabinose DS of only 0.04 was obtained. The removal of arabinose units resulted in the self-assembly of the debranched polymer in water into stable nanoparticle aggregates with a size around 300 nm with a gradual increase in crystallinity of the isolated xylan. Using quartz crystal microbalance with dissipation monitoring, the adsorption of xylan onto model cellulose surfaces was quantified. Compared to the nonmodified xylan, the adsorption of debranched xylan increased from 0.6 to 5.5 mg m(-2). Additionally, adsorption kinetics suggest that the nanoparticles rapidly adsorbed to the cellulose surfaces compared to the arabinoxylan. In summary, a control of the molecular structure of xylan influences its ability to form a new class of polysaccharide nanoparticles in aqueous suspensions and its interaction with nanocellulose surfaces.


Asunto(s)
Nanopartículas/química , Polisacáridos/química , Propiedades de Superficie , Xilanos/química , Adsorción , Biopolímeros/química , Celulosa/química , Nanopartículas/administración & dosificación , Tecnicas de Microbalanza del Cristal de Cuarzo , Soluciones , Agua/química
12.
Appl Microbiol Biotechnol ; 98(17): 7423-35, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24866945

RESUMEN

Bacterial nanocellulose (BNC), synthesized by the bacterium Gluconacetobacter xylinus, is composed of highly hydrated fibrils (99 % water) with high mechanical strength. These exceptional material properties make BNC a novel biomaterial for many potential medical and tissue engineering applications. Recently, BNC with cellulose content of 15 % has been proposed as an implant material for auricular cartilage replacement, since it matches the mechanical requirements of human auricular cartilage. This study investigates the biocompatibility of BNC with increased cellulose content (17 %) to evaluate its response in vitro and in vivo. Cylindrical BNC structures (Ø48 × 20 mm) were produced, purified in a built-in house perfusion system, and compressed to increase the cellulose content in BNC hydrogels. The reduction of endotoxicity of the material was quantified by bacterial endotoxin analysis throughout the purification process. Afterward, the biocompatibility of the purified BNC hydrogels with cellulose content of 17 % was assessed in vitro and in vivo, according to standards set forth in ISO 10993. The endotoxin content in non-purified BNC (2,390 endotoxin units (EU)/ml) was reduced to 0.10 EU/ml after the purification process, level well below the endotoxin threshold set for medical devices. Furthermore, the biocompatibility tests demonstrated that densified BNC hydrogels are non-cytotoxic and cause a minimal foreign body response. In support with our previous findings, this study concludes that BNC with increased cellulose content of 17 % is a promising non-resorbable biomaterial for auricular cartilage tissue engineering, due to its similarity with auricular cartilage in terms of mechanical strength and host tissue response.


Asunto(s)
Celulosa/administración & dosificación , Cartílago Auricular/fisiología , Regeneración Tisular Dirigida/métodos , Hidrogel de Polietilenoglicol-Dimetacrilato/administración & dosificación , Ensayo de Materiales , Animales , Celulosa/metabolismo , Endotoxinas/análisis , Gluconacetobacter xylinus/metabolismo , Hidrogel de Polietilenoglicol-Dimetacrilato/metabolismo , Conejos
13.
Biomed Mater ; 18(4)2023 06 26.
Artículo en Inglés | MEDLINE | ID: mdl-37321229

RESUMEN

Establishing a vascular network in biofabricated tissue grafts is essential for ensuring graft survival. Such networks are dependent on the ability of the scaffold material to facilitate endothelial cell adhesion; however, the clinical translation potential of tissue-engineered scaffolds is hindered by the lack of available autologous sources of vascular cells. Here, we present a novel approach to achieving autologous endothelialisation in nanocellulose-based scaffolds by using adipose tissue-derived vascular cells on nanocellulose-based scaffolds. We used sodium periodate-mediated bioconjugation to covalently bind laminin to the scaffold surface and isolated the stromal vascular fraction and endothelial progenitor cells (EPCs; CD31+CD45-) from human lipoaspirate. Additionally, we assessed the adhesive capacity of scaffold bioconjugationin vitrousing both adipose tissue-derived cell populations and human umbilical vein endothelial cells. The results showed that the bioconjugated scaffold exhibited remarkably higher cell viability and scaffold surface coverage by adhesion regardless of cell type, whereas control groups comprising cells on non-bioconjugated scaffolds exhibited minimal cell adhesion across all cell types. Furthermore, on culture day 3, EPCs seeded on laminin-bioconjugated scaffolds showed positive immunofluorescence staining for the endothelial markers CD31 and CD34, suggesting that the scaffolds promoted progenitor differentiation into mature endothelial cells. These findings present a possible strategy for generating autologous vasculature and thereby increase the clinical relevance of 3D-bioprinted nanocellulose-based constructs.


Asunto(s)
Laminina , Fracción Vascular Estromal , Humanos , Alginatos , Andamios del Tejido , Células Endoteliales de la Vena Umbilical Humana , Ingeniería de Tejidos/métodos
14.
Gels ; 9(10)2023 Oct 12.
Artículo en Inglés | MEDLINE | ID: mdl-37888386

RESUMEN

Autologous fat grafting is hampered by unpredictable outcomes due to high tissue resorption. Hydrogels based on enzymatically pretreated tunicate nanocellulose (ETC) and alginate (ALG) are biocompatible, safe, and present physiochemical properties capable of promoting cell survival. Here, we compared in situ and ex situ crosslinking of ETC/ALG hydrogels combined with lipoaspirate human adipose tissue (LAT) to generate an injectable formulation capable of retaining dimensional stability in vivo. We performed in situ crosslinking using two different approaches; inducing Ca2+ release from CaCO3 microparticles (CMPs) and physiologically available Ca2+ in vivo. Additionally, we generated ex situ-crosslinked, 3D-bioprinted hydrogel-fat grafts. We found that in vitro optimization generated a CMP-crosslinking system with comparable stiffness to ex situ-crosslinked gels. Comparison of outcomes following in vivo injection of each respective crosslinked hydrogel revealed that after 30 days, in situ crosslinking generated fat grafts with less shape retention than 3D-bioprinted constructs that had undergone ex situ crosslinking. However, CMP addition improved fat-cell distribution and cell survival relative to grafts dependent on physiological Ca2+ alone. These findings suggested that in situ crosslinking using CMP might promote the dimensional stability of injectable fat-hydrogel grafts, although 3D bioprinting with ex situ crosslinking more effectively ensured proper shape stability in vivo.

15.
Adv Healthc Mater ; 12(32): e2300640, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37781993

RESUMEN

Intra-portal islet transplantation is currently the only clinically approved beta cell replacement therapy, but its outcome is hindered by limited cell survival due to a multifactorial reaction against the allogeneic tissue in liver. Adipose-derived stromal cells (ASCs) can potentially improve the islet micro-environment by their immunomodulatory action. The challenge is to combine both islets and ASCs in a relatively easy and consistent long-term manner in a deliverable scaffold. Manufacturing the 3D bioprinted double-layered scaffolds with primary islets and ASCs using a mix of alginate/nanofibrillated cellulose (NFC) bioink is reported. The diffusion properties of the bioink and the supportive effect of human ASCs on islet viability, glucose sensing, insulin secretion, and reducing the secretion of pro-inflammatory cytokines are demonstrated. Diabetic mice transplanted with islet-ASC scaffolds reach normoglycemia seven days post-transplantation with no significant difference between this group and the group received islets under the kidney capsules. In addition, animals transplanted with islet-ASC scaffolds stay normoglycemic and show elevated levels of C-peptide compared to mice transplanted with islet-only scaffolds. The data present a functional 3D bioprinted scaffold for islets and ASCs transplanted to the extrahepatic site and suggest a possible role of ASCs on improving the islet micro-environment.


Asunto(s)
Diabetes Mellitus Experimental , Células Secretoras de Insulina , Trasplante de Islotes Pancreáticos , Islotes Pancreáticos , Ratones , Humanos , Animales , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Experimental/metabolismo , Secreción de Insulina , Células Secretoras de Insulina/metabolismo , Células del Estroma/metabolismo , Islotes Pancreáticos/metabolismo , Insulina/metabolismo
16.
Scand Cardiovasc J ; 46(1): 57-62, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22029845

RESUMEN

OBJECTIVES: Many patients in need of bypass surgery lack graft material and current synthetic alternatives have poor performance. A 4 mm vascular graft composed of bacterial cellulose (BC) was developed and tested in pilot study in a large animal model. DESIGN: BC is a biopolymer made by the bacteria acetobacter xylinum. BC grafts (n = 16) with 4 cm length and 4 mm internal diameter were implanted bilaterally in the carotid arteries of eight sheep. No long-term antithrombotic therapy was administered. Patency was assessed with ultrasound. Histology, immunohistochemistry, and electron microscopy were performed after explantation. RESULTS: Fifty percent of the grafts occluded within two weeks. One animal died with patent grafts after 14 days. In the three remaining animals 5/6 grafts were patent after nine months. Two animals were followed 13 months after implantation with 3/4 grafts patent at explantation. All patent grafts had confluent endothelial-like cells. CONCLUSIONS: Biosynthetic small calibre vascular grafts made from BC can be patent for up to 13 months in sheep carotid arteries. BC is a potential material for small calibre grafts but patency in animal models needs to be improved before clinical studies can be planned.


Asunto(s)
Prótesis Vascular , Celulosa , Animales , Arterias Carótidas/patología , Arterias Carótidas/cirugía , Arterias Carótidas/ultraestructura , Celulosa/metabolismo , Células Endoteliales/patología , Endotelio Vascular/fisiología , Gluconacetobacter xylinus/metabolismo , Oclusión de Injerto Vascular/patología , Modelos Animales , Ovinos , Grado de Desobstrucción Vascular/fisiología
17.
Adipocyte ; 11(1): 34-46, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-34957918

RESUMEN

Three-dimensional (3D)-bioprinted lipoaspirate-derived adipose tissue (LAT) is a potential alternative to lipo-injection for correcting soft-tissue defects. This study investigated the long-term in vivo survival of 3D-bioprinted LAT and its proteomic signature and cellular composition. We performed proteomic and multicolour flow cytometric analyses on the lipoaspirate and 3D-bioprinted LAT constructs were transplanted into nude mice, followed by explantation after up to 150 days. LAT contained adipose-tissue-derived stem cells (ASCs), pericytes, endothelial progenitor cells (EPCs) and endothelial cells. Proteomic analysis identified 6,067 proteins, including pericyte markers, adipokines, ASC secretome proteins, proangiogenic proteins and proteins involved in adipocyte differentiation and developmental morphogenic signalling, as well as proteins not previously described in human subcutaneous fat. 3D-bioprinted LAT survived for 150 days in vivo with preservation of the construct shape and size. Furthermore, we identified human blood vessels after 30 and 150 days in vivo, indicating angiogenesis from capillaries. These results showed that LAT has a favourable proteomic signature, contains ASCs, EPCs and blood vessels that survive 3D bioprinting and can potentially facilitate angiogenesis and successful autologous fat grafting in soft-tissue reconstruction.


Asunto(s)
Células Progenitoras Endoteliales , Proteómica , Tejido Adiposo/metabolismo , Animales , Humanos , Ratones , Ratones Desnudos , Secretoma
18.
Carbohydr Polym ; 286: 119284, 2022 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-35337506

RESUMEN

Alginate has been used for decades for cell encapsulation. Cellulose nanofibrils (CNF) from tunicates are desirable in biomedicine due to high molecular weight, purity, crystallinity, and sustainable production. We prepared microbeads of 400-600 µm of alginate and tunicate CNF. Greater size, dispersity and aspect ratio were observed in microbeads with higher fractions of CNF. CNF content in Ca-crosslinked alginate microbeads decreased stability upon saline exposure, whereas crosslinking with calcium (50 mM) and barium (1 mM) yielded stable microbeads. The Young's moduli of gel cylinders decreased when exchanging alginate with CNF, and slightly increased permeability to dextran was observed in microbeads containing CNF. Encapsulation of MC3T3 cells revealed high cell viability after encapsulation (83.6 ± 0.4%) in beads of alginate and CNF. NHDFs showed lower viability but optimizing mixing and production techniques of microbeads increased cell viability (from 66.2 ± 5.3% to 72.7 ± 7.5%).


Asunto(s)
Alginatos , Urocordados , Animales , Encapsulación Celular , Celulosa/farmacología , Microesferas
19.
Biomater Adv ; 137: 212828, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35929261

RESUMEN

Extracellular matrix fibril components, such as collagen, are crucial for the structural properties of several tissues and organs. Tunicate-derived cellulose nanofibrils (TNC) combined with living cells could become the next gold standard for cartilage and soft-tissue repair, as TNC fibrils present similar dimensions to collagen, feasible industrial production, and chemically straightforward and cost-efficient extraction procedures. In this study, we characterized the physical properties of TNC derived from aquaculture production in Norwegian fjords and evaluated its biocompatibility regarding induction of an inflammatory response and foreign-body reactions in a Wistar rat model. Additionally, histologic and immunohistochemical analyses were performed for comparison with expanded polytetrafluoroethylene (ePTFE) as a control. The average length of the TNC as determined by atomic force microscopy was tunable from 3 µm to 2.4 µm via selection of a various number of passages through a microfluidizer, and rheologic analysis showed that the TNC hydrogels were highly shear-thinning and with a viscosity dependent on fibril length and concentration. As a bioink, TNC exhibited excellent rheological and printability properties, with constructs capable of being printed with high resolution and fidelity. We found that post-print cross-linking with alginate stabilized the construct shape and texture, which increased its ease of handling during surgery. Moreover, after 30 days in vivo, the constructs showed a highly-preserved shape and fidelity of the grid holes, with these characteristics preserved after 90 days and with no signs of necrosis, infection, acute inflammation, invasion of neutrophil granulocytes, or extensive fibrosis. Furthermore, we observed a moderate foreign-body reaction involving macrophages, lymphocytes, and giant cells in both the TNC constructs and PTFE controls, although TNC was considered a non-irritant biomaterial according to ISO 10993-6 as compared with ePTFE. These findings represent a milestone for future clinical application of TNC scaffolds for tissue repair. One sentence summary: In this study, the mechanical properties of tunicate nanocellulose are superior to nanocellulose extracted from other sources, and the biocompatibility is comparable to that of ePTFE.


Asunto(s)
Ingeniería de Tejidos , Urocordados , Animales , Materiales Biocompatibles/química , Celulosa/farmacología , Colágeno/farmacología , Ratas , Ratas Wistar , Ingeniería de Tejidos/métodos
20.
Langmuir ; 27(14): 8718-28, 2011 Jul 19.
Artículo en Inglés | MEDLINE | ID: mdl-21699205

RESUMEN

Adsorption of anionic polyelectrolytes, sodium salts of carboxymethyl celluloses (CMCs) with different degrees of substitution (DS = 0.9 and 1.2), from aqueous electrolyte solutions onto regenerated cellulose surfaces was studied using quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) experiments. The influence of both calcium chloride (CaCl(2)) and sodium chloride (NaCl) on CMC adsorption was examined. The QCM-D results demonstrated that CaCl(2) (divalent cation) caused significantly greater CMC adsorption onto regenerated cellulose surfaces than NaCl (monovalent cation) at the same ionic strength. The CMC layers adsorbed onto regenerated cellulose surfaces from CaCl(2) solutions exhibited greater stability upon exposure to flowing water than layers adsorbed from NaCl solutions. Both QCM-D and SPR results showed that CMC adsorption onto regenerated cellulose surfaces from CaCl(2) solutions increased with increasing CaCl(2) concentration up to the solubility limit (10 mM). Voigt-based viscoelastic modeling of the QCM-D data indicated that the CMC layers adsorbed onto regenerated cellulose surfaces had shear viscosities of η(f) ≈ 10(-3) N·s·m(-2) and elastic shear moduli of µ(f) ≈ 10(5) N·m(-2). Furthermore, the combination of SPR spectroscopy and QCM-D showed that the CMC layers contained 90-95% water. Adsorption isotherms for CMCs in CaCl(2) solutions were also obtained from QCM-D and were fit by Freundlich isotherms. This study demonstrated that CMC adsorption from CaCl(2) solutions is useful for the modification of cellulose surfaces.


Asunto(s)
Carboximetilcelulosa de Sodio/química , Tecnicas de Microbalanza del Cristal de Cuarzo , Resonancia por Plasmón de Superficie , Adsorción , Cloruro de Calcio/química , Catálisis , Gases/química , Ácido Clorhídrico/química , Polímeros/química , Sales (Química)/química , Cloruro de Sodio/química , Propiedades de Superficie , Sustancias Viscoelásticas/química
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