The decellularized extracellular matrix (ECM) obtained from human and porcine adipose tissue (AT) is currently used to prepare regenerative medicine bio-scaffolds. However, the influence of these natural biomaterials on host immune response is not yet deeply understood. Since macrophages play a key role in the inflammation/healing processes due to their high functional plasticity between M1 and M2 phenotypes, the evaluation of their response to decellularized ECM is mandatory. It is also necessary to analyze the immunocompetence of macrophages after contact with decellularized ECM materials to assess their functional role in a possible infection scenario. In this work, we studied the effect of four decellularized adipose matrices (DAMs) obtained from human and porcine AT by enzymatic or chemical methods on macrophage phenotypes and fungal phagocytosis. First, a thorough biochemical characterization of these biomaterials by quantification of remnant DNA, lipids, and proteins was performed, thus indicating the efficiency and reliability of both methods. The proteomic analysis evidenced that some proteins are differentially preserved depending on both the AT origin and the decellularization method employed. After exposure to the four DAMs, specific markers of M1 proinflammatory and M2 anti-inflammatory macrophages were analyzed. Porcine DAMs favor the M2 phenotype, independently of the decellularization method employed. Finally, a sensitive fungal phagocytosis assay allowed us to relate the macrophage phagocytosis capability with specific proteins differentially preserved in certain DAMs. The results obtained in this study highlight the close relationship between the ECM biochemical composition and the macrophage’s functional role.
survival was demonstrated in all the membranes after three months follow-up. A slight reduction in the extrusion rate of h-ADASC colonized materials was observed. No significant differences between the groups with and without h-ADASC were detected respect to transparency or neovascularization. We propose PEA with low hydroxylation as a scaffold for the anchoring ring of future keratoprosthesis.
A simple and economic method is presented that allows the preparation of transparent polystyrene (PS) substrates activated with chlorosulfonyl groups. Chlorosulfonation has been analyzed by ATR-FTIR. Linear PS chains with different degrees of chlorosulfonation have been synthesized as model compounds in order to analyze the modification quantitatively. After chlorosulfonation the activated surfaces can be quantitatively converted in aqueous solution at room temperature to sulfo or sulfonazide groups or react with bifunctional aliphatic amines of different length via formation of sulfonamide linkages. In this way, surfaces with a huge variety of functionalities like amines, carboxylic or sulfonic groups, sulfonazides, esters, etc. may be obtained in a selective way controlling their density at the surface. In all cases, functional surfaces with excellent optical transparency are obtained. Aminated surfaces have successfully been probed for ELISA assays.
Biomaterials with surface antibacterial properties are promising components for medical implants that might provide an alternative to conventional systemic antibiotic treatments. Herein is reported a general method, based on plasma polymerization techniques, to promote the formation of “clickable surfaces” which can be conjugated with chemically modified antibiotics (e.g., azido‐vancomycin) under very mild conditions. The procedure is comprised of three operations: (i) surface alkylcarboxylation with acrylic acid/CO2 plasma, (ii) alkyne functionalization by condensation with propargylamine, and (iii) in situ Cu(I)‐catalyzed alkyne–azide conjugation with azidovancomycin. The antibacterial activity of the resulting functionalized surfaces has been assessed against Staphylococcus epidermidis.
3D cell culture systems based on biological scaffold materials obtainable from both animal and human tissues constitute very interesting tools for cell therapy and personalised medicine applications. The white adipose tissue (AT) extracellular matrix (ECM) is a very promising biomaterial for tissue engineering due to its easy accessibility, malleability and proven biological activity. In the present study, human dental pulp stem cells (hDPSCs) were combined in vitro with ECM scaffolds from porcine and human decellularised adipose tissues (pDAT, hDAT) processed as 3D solid foams, to investigate their effects on the osteogenic differentiation capacity and bone matrix production of hDPSCs, compared to single-protein-based 3D solid foams of collagen type I and conventional 2D tissue-culture-treated polystyrene plates. pDAT solid foams supported the osteogenic differentiation of hDPSCs to similar levels to collagen type I, as assessed by alkaline phosphatase and alizarin red stainings, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and osteocalcin/bone gamma-carboxyglutamate protein (BGLAP) immunostaining. Interestingly, hDAT solid foams showed a markedly lower capacity to sustain hDPSC osteogenic differentiation and matrix calcification and a higher capacity to support adipogenesis, as assessed by RT-qPCR and oil red O staining. White ATs from both human and porcine origins are relatively abundant and available sources of raw material to obtain high quality ECM-derived biomedical products. These biomaterials could have promising applications in tissue engineering and personalised clinical therapy for the healing and regeneration of lesions involving not only a loss of calcified bone but also its associated soft non-calcified tissues.
In this work the activation of transparent PS substrates by chlorosulfonation is described and their distribution in the subsurface region is analyzed. For this purpose XPS, FTIR-ATR and colorimetry have been used. It is shown that the electrophilic aromatic substitution of polystyrene in pure chlorosulfonic acid is extremely quick with complete surface coverage by chlorosulfonic groups achieved after only a 10 minute reaction time at -10 °C. It is further demonstrated that the reaction is very surface selective and that even after reaction times as long as 3 hours, the modification is limited to a layer with a thickness of less than one micron. The activated PS substrates can be further functionalized in a second step with carboxylic groups. Due to the excellent optical transparency that the samples maintain upon modification, the modified systems were successfully probed for use in ELISA assays.
Synthesis of atomic nanoclusters (NCs) using proteins as scaffold has attracted great attention. Usually the synthetic conditions for the synthesis of NCs stabilized with proteins require extreme pHs or temperature.These harsh reaction conditions cause the denaturation of the proteins and end up in the loss of their biological functions. Until now there are no examples of the use of antibodies as NCs stabilisers. In this work we present the first method for the synthesis of catalytic NCs that uses antibodies for the stabilization of NCs. Anti-BSA IgG was used as a model in order to demonstrate that it is possible to use an antibody as scaffold for the synthesis of semiconductor and metallic NCs with catalytic properties. The synthesis of antibodies modified with NCs is carried out under non denaturing conditions, which do not affect the antibody structure. The resulting antibodies still maintain the affinity for target antigens and protein G. The catalytic properties of the anti-BSA IgG modified with NCs can be used to the quantification of BSA in a direct sandwich ELISA.Supporting Information. Experimental section, absorption and fluorescence spectra of CdS NCs-IgG and Ag/Pt NCs-IgG, ICP analysis, optimization experiments, characterization of BSA-stabilized NCs, and size distribution of NCs. The following files are available free of charge.
Locally applied antibiotics under temporally controlled release present many advantages over systemic clinical treatments, e.g. efficiency and side effects. This can be achieved by a coating on top of the medical device, in which the antibiotic is stored. This study presents the use of plasma polymerization to produce such a coating using N,O‐bis‐tert‐butyldimethylsilylated ciprofloxacin (silylciprofloxacin) as a precursor. Once exposed to physiological media, the outer layers of the coating release the antibiotic by a hydrolysis reaction. Thus, the plasma process parameters can control the speed of liberation through the coating polymerization. Besides, this study shows that the release products present antibiotic activity against a number of bacteria: E. coli, P. aeruginosa, and S. aureus.
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