Determinants of diversity and composition of the tapeworm fauna of blue sharks, Prionace glauca: a geographical and host-specificity analysis.

Blue sharks, Prionace glauca, are cosmopolitan, extremely vagile sharks and the species among elasmobranchs for which most surveys containing tapeworm community data are available worldwide. In this study we report on the tapeworm fauna of three samples of blue sharks (n = 37) from two new regions (one sample from Galicia, north-east Atlantic, and two from Valencia, western Mediterranean), and compared it with previous studies, assessing the relative role of the ecological and evolutionary factors in structuring local tapeworm assemblages. Nine cestode taxa were identified, of which four included adult specimens, that is, Platybothrium auriculatum, Prosobothrium armigerum, Anthobothrium caseyi and Molicola horridus. The abundance of these species, and Brillouin's diversity index, differed significantly among samples without a clear geographical signal. A comparison with six previous surveys revealed that tapeworm assemblages were composed of the same 'core' taxa, with mean species richness typically ranging from two to four species. Global records of adult tapeworms in blue sharks included: 15 taxa identified at species level, of which only eight (generalist trypanorhynchs) were shared with other sympatric host species; five mostly with other carcharhinids; and three with large lamnid sharks sharing the blue sharks' habitat. The composition of tapeworm communities of blue sharks is thus highly constrained by strong host specificity, with composition and abundance varying across localities depending on idiosyncratic environmental conditions.

Correlation between biological responses in vitro and in vivo to Ca-doped sol-gel coatings assessed using proteomic analysis.

Poor correlation between the results of in vitro testing and the subsequent in vivo experiments hinders the design of biomaterials. Thus, new characterisation methods are needed. This study used proteomic and histological techniques to analyse the effects of Ca-doped biomaterials in vitro and in vivo and verify the correlation between the two systems. The sol-gel route was employed to synthesise coatings functionalised with 0.5 and 5 wt% of CaCl2. Morphology of the coatings was examined using SEM; the Ca2+ ion release from the materials was analysed by means of ICP-AES spectroscopy. The osteogenic and inflammatory responses were inspected in vitro in human osteoblasts (HOb) and TPH-1 monocytes. The in vivo experiments used a rabbit model. The nLC-MS/MS-based proteomic methods were utilised to analyse the proteins adhering to the material samples incubated with human serum or examine protein expression in the tissues close to the implants. Ca-doped biomaterials caused a remarkable increase in the adsorption of coagulation-related proteins, both in vitro (PLMN, THRB, FIBA and VTNC) and in vivo (FBLN1, G1U978). Enhanced affinity to these materials was also observed for proteins involved in inflammation (CO5, C4BPA, IGHM and KV302 in vitro; CARD6, DDOST and CD14 in vivo) and osteogenic functions (TETN, PEDF in vitro; FBN1, AHSG, MYOC in vivo). The results obtained using different techniques were well matched, with a good correlation between the in vitro and in vivo experiments. Thus, the proteomic analysis of biological responses to biomaterials in vitro is a useful tool for predicting their impact in vivo.

Bioactive zinc-doped sol-gel coating modulates protein adsorption patterns and in vitro cell responses.

Zinc is an essential element with an important role in stimulating the osteogenesis and mineralization and suppressing osteoclast differentiation. In this study, new bioactive ZnCl2-doped sol-gel materials were designed to be applied as coatings onto titanium. The biomaterials were physicochemically characterized and the cellular responses evaluated in vitro using MC3T3-E1 osteoblasts and RAW264.7 macrophages. The effect of Zn on the adsorption of human serum proteins onto the material surface was evaluated through nLC-MS/MS. The incorporation of Zn did not affect the crosslinking of the sol-gel network. A controlled Zn2+ release was obtained, reaching values below 10 ppm after 21 days. The materials were no cytotoxic and lead to increased gene expression of ALP, TGF-ß, and RUNX2 in the osteoblasts. In macrophages, an increase of IL-1ß, TGF-ß, and IL-4 gene expression was accompanied by a reduced TNF-α liberation. Proteomic results showed changes in the adsorption patterns of proteins associated with immunological, coagulative, and regenerative functions, in a Zn dose-dependent manner. The variations in protein adsorption might lead to the downregulation of the NF-κB pathway, thus explain the observed biological effects of Zn incorporation into biomaterials. Overall, these coatings demonstrated their potential to promote bone tissue regeneration.

Complement proteins regulating macrophage polarisation on biomaterials.

One of the events occurring when a biomaterial is implanted in an host is the protein deposition onto its surface, which might regulate cell responses. When a biomaterial displays a compromised biocompatibility, distinct complement pathways can be activated to produce a foreign body reaction. In this article, we have designed different types of biomaterial surfaces to study the inflammation process. Here, we used different concentrations of (3-glycidoxypropyl)-trimethoxysilane (GPTMS), an organically-modified alkoxysilane as a precursor for the synthesis of various types of sol-gel materials functionalizing coatings for titanium implants to regulate biological responses. Our results showed that greater GPTMS surface concentrations induced greater secretion of TNF-α and IL-10 on RAW 264.7 macrophages. When implanted into rabbit tibia, osseointegration decreased with higher GPTMS concentrations. Interestingly, higher deposition of complement-related proteins C-reactive protein (CRP) and ficolin-2 (FCN2), two main activators of distinct complement pathways, was observed. Taking all together, inflammatory potential increase seems to be GPTMS concentration-dependent. Our results show that a greater adsorption of complement proteins can condition macrophage polarization.

Proteomic analysis of calcium-enriched sol-gel biomaterials.

Calcium is an element widely used in the development of biomaterials for bone tissue engineering as it plays important roles in bone metabolism and blood coagulation. The Ca ions can condition the microenvironment at the tissue-material interface, affecting the protein deposition process and cell responses. The aim of this study was to analyze the changes in the patterns of protein adsorption on the silica hybrid biomaterials supplemented with different amounts of CaCl2, which can function as release vehicles. This characterization was carried out by incubating the Ca-biomaterials with human serum. LC-MS/MS analysis was used to characterize the adsorbed protein layers and compile a list of proteins whose affinity for the surfaces might depend on the CaCl2 content. The attachment of pro- and anti-clotting proteins, such as THRB, ANT3, and PROC, increased significantly on the Ca-materials. Similarly, VTNC and APOE, proteins directly involved on osteogenic processes, attached preferentially to these surfaces. To assess correlations with the proteomic data, these formulations were tested in vitro regarding their osteogenic and inflammatory potential, employing MC3T3-E1 and RAW 264.7 cell lines, respectively. The results confirmed a Ca dose-dependent osteogenic and inflammatory behavior of the materials employed, in accordance with the protein attachment patterns.

The effect of strontium incorporation into sol-gel biomaterials on their protein adsorption and cell interactions.

It is known strontium can both inhibit the osteoclast formation and stimulate the osteoblast maturation, so biomaterials containing this element can favour bone structure stabilisation. The addition of Sr to biomaterials could affect their interactions with proteins and cells. Here, a silica-hybrid sol-gel network doped with different amounts of SrCl2 and applied as coatings on titanium discs was examined. in vitro analysis was performed to determine the potential effect of Sr in the coatings, showing enhanced gene expression of osteogenic markers (alkaline phosphatase and transforming growth factor-ß) in MC3T3-E1 incubated with Sr-doped biomaterials. The examination of inflammatory markers (tumour necrosis factor-α and interleukin 10) in RAW 264.7 macrophages revealed an anti-inflammatory potential of these materials. Proteins adsorbed onto the coatings incubated with human serum (3 h at 37 °C) were also analysed; mass spectrometry was used to characterise the proteins adhering to materials with different Sr content. Adding Sr to the coatings increased their affinity to APOE and VTNC proteins (associated with anti-inflammatory and osteogenic functions). Moreover, the proteins involved in coagulation processes, such as prothrombin, were more abundant on the coatings containing Sr than on the base sol-gel surfaces. Correlations between gene expression and proteomic results were also examined.

Synthesis and characterization of silica-chitosan hybrid materials as antibacterial coatings for titanium implants.

To avoid dental implant-related infections and to promote the osseointegration of titanium implants, the application of silicon and chitosan containing coatings is proposed. Silicon is a well-known osteogenic element and chitosan was selected to confer the antibacterial properties. The synthesis of hybrid silica-chitosan coatings using the sol-gel process is presented and the characterization using 29Si-NMR to verify the correct formation of the network is discussed. The 13C NMR spectroscopy was used to confirm the covalent union between chitosan and the silicon network. Hydrolytic degradation and silicon release studies showed the effective silicon release from the hybrids and, hence, the possibility to promote bone formation. The introduction of different amounts of chitosan and tetraethyl orthosilicate (TEOS) modulated the Si release. The analysis of cell cultures in vitro demonstrated that the hybrid coatings were not cytotoxic and promoted cell proliferation on their surfaces. The coatings containing 5%-10% chitosan had substantial antibacterial properties.

Osseointegration mechanisms: a proteomic approach.

The prime objectives in the development of biomaterials for dental applications are to improve the quality of osseointegration and to short the time needed to achieve it. Design of implants nowadays involves changes in the surface characteristics to obtain a good cellular response. Incorporating osteoinductive elements is one way to achieve the best regeneration possible post-implantation. This study examined the osteointegrative potential of two distinct biomaterials: sandblasted acid-etched titanium and a silica sol-gel hybrid coating, 70% MTMOS-30% TEOS. In vitro, in vivo, and proteomic characterisations of the two materials were conducted. Enhanced expression levels of ALP and IL-6 in the MC3T3-E1 cells cultured with coated discs, suggest that growing cells on such surfaces may increase mineralisation levels. 70M30T-coated implants showed improved bone growth in vivo compared to uncoated titanium. Complete osseointegration was achieved on both. However, coated implants displayed osteoinductive properties, while uncoated implants demonstrated osteoconductive characteristics. Coagulation-related proteins attached predominantly to SAE-Ti surface. Surface properties of the material might drive the regenerative process of the affected tissue. Analysis of the proteins on the coated dental implant showed that few proteins specifically attached to its surface, possibly indicating that its osteoinductive properties depend on the silicon delivery from the implant.

Bioactive potential of silica coatings and its effect on the adhesion of proteins to titanium implants.

There is an ever-increasing need to develop dental implants with ideal characteristics to achieve specific and desired biological response in the scope of improve the healing process post-implantation. Following that premise, enhancing and optimizing titanium implants through superficial treatments, like silica sol-gel hybrid coatings, are regarded as a route of future research in this area. These coatings change the physicochemical properties of the implant, ultimately affecting its biological characteristics. Sandblasted acid-etched titanium (SAE-Ti) and a silica hybrid sol-gel coating (35M35G30T) applied onto the Ti substrate were examined. The results of in vitro and in vivo tests and the analysis of the protein layer adsorbed to each surface were compared and discussed. In vitro analysis with MC3T3-E1 osteoblastic cells, showed that the sol-gel coating raised the osteogenic activity potential of the implants (the expression of osteogenic markers, the alkaline phosphatase (ALP) and IL-6 mRNAs, increased). In the in vivo experiments using as model rabbit tibiae, both types of surfaces promoted osseointegration. However, the coated implants demonstrated a clear increase in the inflammatory activity in comparison with SAE-Ti. Mass spectrometry (LC-MS/MS) analysis showed differences in the composition of protein layers formed on the two tested surfaces. Large quantities of apolipoproteins were found attached predominantly to SAE-Ti. The 35M35G30T coating adsorbed a significant quantity of complement proteins, which might be related to the material intrinsic bioactivity, following an associated, natural and controlled immune response. The correlation between the proteomic data and the in vitro and in vivo outcomes is discussed on this experimental work.

Proteomic analysis of silica hybrid sol-gel coatings: a potential tool for predicting the biocompatibility of implants in vivo.

The interactions of implanted biomaterials with the host organism determine the success or failure of an implantation. Normally, their biocompatibility is assessed using in vitro tests. Unfortunately, in vitro and in vivo results are not always concordant; new, effective methods of biomaterial characterisation are urgently needed to predict the in vivo outcome. As the first layer of proteins adsorbed onto the biomaterial surfaces might condition the host response, mass spectrometry analysis was performed to characterise these proteins. Four distinct hybrid sol-gel biomaterials were tested. The in vitro results were similar for all the materials examined here. However, in vivo, the materials behaved differently. Six of the 171 adsorbed proteins were significantly more abundant on the materials with weak biocompatibility; these proteins are associated with the complement pathway. Thus, protein analysis might be a suitable tool to predict the in vivo outcomes of implantations using newly formulated biomaterials.

Osteomielitis crónica multifocal recurrente / Chronic recurrent multifocal osteomyelitis

La osteomielitis crónica multifocal recurrente se considera actualmente una variante de un trastorno autoinflamatorio infrecuente, que requiere un alto índice de sospecha clínica para efectuar un adecuado abordaje diagnóstico y terapéutico. Presentamos un caso clínico típico, aportando datos sobre las hipótesis etiopatogénicas que se manejan actualmente en esta entidad (AU)

Chronic recurrent multifocal osteomyelitis is currently considered a variant of a rare autoinflammatory disorder, which requires a high index of clinic suspicion for suitable diagnostic and therapeutic approach. We present a typical case, providing data on etiopathogenetic hypothesis currently handled in this entity. The early detection of these malformations can prevent the appearance of chronic lung diseases and can allow for the best therapeutic approach. Although most of the cases do not require treatment, some of them require specific and more aggressive handling (AU)

Synthesis of hybrid sol-gel materials and their biological evaluation with human mesenchymal stem cells.

Surface engineering of biomaterials could promote the osseointegration of implants. In this work, two types of hybrid sol-gel materials were developed to stimulate cell attachment, proliferation and differentiation of osteogenic cells. One type was synthesised from vinyl triethoxysilane (VTES) and tetraethyl-orthosilicate (TEOS) at different molar ratios, while the other from VTES and hydroxyapatite particles (HAp). Hybrid materials were systematically investigated using nuclear magnetic resonance, Fourier transform infrared spectroscopy and contact angle metrology. The biocompatibility and osseoinduction of the coatings were evaluated by measuring mesenchymal stem cell proliferation using MTT assays and analysing the mineralised extracellular matrix production by quantifying calcium-rich deposits. The results highlighted the versatility of these coatings in obtaining different properties by changing the molar ratio of the VTES:TEOS precursors. Thus, mineralisation was stimulated by increasing TEOS content, while the addition of HAp improved cell proliferation but worsened mineralisation.

Biodegradable PCL scaffolds with an interconnected spherical pore network for tissue engineering.

A technique for producing controlled interconnected porous structures for application as a tissue engineering scaffold is presented in this article. The technique is based on the fabrication of a template of interconnected poly(ethyl methacrylate) (PEMA) microspheres, the introduction of a biodegradable polymer, poly-epsilon-caprolactone (PCL), and the elimination of the template by a selective solvent. A series of PCL scaffolds with a porosity of 70% and pore sizes up to 200 microm were produced and characterized (both thermally and mechanically). Human chondrocytes were cultured in monolayer on bulk PCL disks or seeded into porous PCL scaffolds. Cell adhesion, viability, proliferation, and proteoglycan (PG) synthesis were tested and compared with monolayer cultures on tissue-treated polystyrene or pellet cultures as reference controls. Cells cultured on PCL disks showed an adhesion similar to that of the polystyrene control (which allowed high levels of proliferation). Stained scaffold sections showed round-shaped chondrocyte aggregates embedded into porous PCL. PG production was similar to that of the pellet cultures and higher than that obtained with monolayer postconfluence cultures. This shows that the cells are capable of attaching themselves to PCL. Furthermore, in porous PCL, cells maintain the same phenotype as the chondrocytes within the native cartilage. These results suggest that PCL scaffolds may be a suitable candidate for chondrocyte culture.

A porous PCL scaffold promotes the human chondrocytes redifferentiation and hyaline-specific extracellular matrix protein synthesis.

The redifferentiation, proliferation, and hyaline-specific extracellular matrix (ECM) protein synthesis of chondrocytes cultured in a polycaprolactone (PCL) scaffold were analyzed. Gene expression of the type II collagen and aggrecan was assessed by real-time PCR in cells from PCL scaffolds, monolayer, and pellet cultures. The proliferative activity was assessed using Ki-67 immunodetection, and the chondrocytic differentiation was evaluated using S-100 immunodetection. The synthesis and deposition into scaffold pores of type II collagen and glycosaminoglycan were analyzed by immunohistochemistry and Alcian blue staining, respectively. All parameters were assessed throughout 28 days of cultures maintained in either fetal bovine serum-containing medium (FCM) or Insulin-Transferrin-Selenium-containing medium (ICM). Expression of the type II collagen gene was lower in FCM cultures than in ICM cultures for all culture systems (p < 0.05). Moreover, PCL scaffolds cultured in ICM were able to induce collagen gene expression more efficiently than pellet and monolayer cultures. Aggrecan gene expression did not vary significantly between mediums and three-dimensional system cultures, but in ICM cultures, the monolayer cultures had significantly higher levels of aggrecan gene expression than did either the PCL or pellet cultures. Chondrocytes cultured in PCL scaffolds or pellets with FCM did not proliferate to a great extent but did maintain their differentiated phenotype for 28 days. Levels of cartilage ECM protein synthesis and deposition into the scaffold pores were similar among PCL and pellet cultures grown in FCM and in ICM. In conclusion, chondrocytes seeded into PCL scaffolds, cultured in ICM, efficiently maintained their differentiated phenotype and were able to synthesize cartilage-specific ECM proteins.