Junctional epithelium and hemidesmosomes: Tape and rivets for solving the “percutaneous device dilemma” in dental and other permanent implants

The percutaneous device dilemma describes etiological factors, centered around the disrupted epithelial tissue surrounding non-remodelable devices, that contribute to rampant percutaneous device infection. Natural percutaneous organs, in particular their extracellular matrix mediating the “device”/epithelium interface, serve as exquisite examples to inspire longer lasting long-term percutaneous device design. For example, the tooth's imperviousness to infection is mediated by the epithelium directly surrounding it, the junctional epithelium (JE). The hallmark feature of JE is formation of hemidesmosomes, cell/matrix adhesive structures that attach surrounding oral gingiva to the tooth's enamel through a basement membrane. Here, the authors survey the multifaceted functions of the JE, emphasizing the role of the matrix, with a particular focus on hemidesmosomes and their five main components. The authors highlight the known (and unknown) effects dental implant – as a model percutaneous device – placement has on JE regeneration and synthesize this information for application to other percutaneous devices. The authors conclude with a summary of bioengineering strategies aimed at solving the percutaneous device dilemma and invigorating greater collaboration between clinicians, bioengineers, and matrix biologists. © 2022 The Author

Intrafibrillar Mineralization of Self-Assembled Elastin-Like Recombinamer Fibrils

Producción CientíficaBiomineralization of bone, a controlled process where hydroxyapatite nanocrystals preferentially deposit in collagen fibrils, is achieved by the interplay of the collagen matrix and noncollagenous proteins. Mimicking intrafibrillar mineralization in synthetic systems is highly attractive for the development of advanced hybrid materials with elaborated morphologies and outstanding mechanical properties, as well as understanding the mechanisms of biomineralization. Inspired by nature, intrafibrillar mineralization of collagen fibrils has been successfully replicated in vitro via biomimetic systems, where acidic polymeric additives are used as analogue of noncollagenous proteins in mediating mineralization. The development of synthetic templates that mimic the structure and functions of collagenous matrix in mineralization has yet to be explored. In this study, we demonstrated that self-assembled fibrils of elastin-like recombinamers (ELRs) can induce intrafibrillar mineralization. The ELRs displayed a disordered structure at low temperature but self-assembled into nanofibrils above its inverse transition temperature. In the presence of the self-assembled ELR fibrils, polyaspartate-stabilized amorphous calcium phosphates preferentially infiltrated into the fibrils and then crystallized into hydroxyapatite nanocrystals with their [001] axes aligned parallel to the long axis of the ELR fibril. As the recombinant technology enables designing and producing well-defined ELRs, their molecular and structural properties can be fine-tuned. By examining the ultrastructure of the self-assembled ELRs fibrils as well as their mineralization, we concluded that the spatial confinement formed by a continuum β-spiral structure in an unperturbed fibrillar structure rather than electrostatic interactions or bioactive sequences in the recombinamer composition played the crucial role in inducing intrafibrillar mineralization.2018-08-01Ministerio de Economía, Industria y Competitividad (Project MAT2013-42473-R and MAT2015-68901R)Junta de Castilla y León (programa de apoyo a proyectos de investigación – Ref. VA244U13, VA313U14 and VA015U16

Implant surface physicochemistry affects keratinocyte hemidesmosome formation

Previous studies have shown hydrophilic/hydrophobic implant surfaces stimulate/hinder osseointegration. An analogous concept was applied here using common biological functional groups on a model surface to promote oral keratinocytes (OKs) proliferation and hemidesmosomes (HD) to extend implant lifespans through increased soft tissue attachment. However, it is unclear what physicochemistry stimulates HDs. Thus, common biological functional groups (NH2 , OH, and CH3 ) were functionalized on glass using silanization. Non-functionalized plasma-cleaned glass and H silanization were controls. Surface modifications were confirmed with X-ray photoelectron spectroscopy and water contact angle. The amount of bovine serum albumin (BSA) and fibrinogen, and BSA thickness, were assessed to understand how adsorbed protein properties were influenced by physicochemistry and may influence HDs. OKs proliferation was measured, and HDs were quantified with immunofluorescence for collagen XVII and integrin β4. Plasma-cleaned surfaces were the most hydrophilic group overall, while CH3 was the most hydrophobic and OH was the most hydrophilic among functionalized groups. Modification with the OH chemical group showed the highest OKs proliferation and HD expression. The OKs response on OH surfaces appeared to not correlate to the amount or thickness of adsorbed model proteins. These results reveal relevant surface physicochemical features to favor HDs and improve implant soft tissue attachment.© 2023 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC

Immunomodulatory IL-23 receptor antagonist peptide nanocoatings for implant soft tissue healing

Peri-implantitis, caused by an inflammatory response to pathogens, is the leading cause of dental implant failure. Poor soft tissue healing surrounding implants - caused by inadequate surface properties - leads to infection, inflammation, and dysregulated keratinocyte and macrophage function. One activated inflammatory response, active around peri-implantitis compared to healthy sites, is the IL-23/IL-17A cytokine axis. Implant surfaces can be synthesized with peptide nanocoatings to present immunomodulatory motifs to target peri-implant keratinocytes to control macrophage polarization and regulate inflammatory axises toward enhancing soft tissue healing.We synthesized an IL-23 receptor (IL-23R) noncompetitive antagonist peptide nanocoating using silanization and evaluated keratinocyte secretome changes and macrophage polarization (M1-like "pro-inflammatory" vs. M2-like "pro-regenerative").IL-23R antagonist peptide nanocoatings were successfully synthesized on titanium, to model dental implant surfaces, and compared to nonfunctional nanocoatings and non-coated titanium. IL-23R antagonist nanocoatings significantly decreased keratinocyte IL-23, and downstream IL-17A, expression compared to controls. This peptide noncompetitive antagonistic function was demonstrated under lipopolysaccharide stimulation. Large scale changes in keratinocyte secretome content, toward a pro-regenerative milieu, were observed from keratinocytes cultured on the IL-23R antagonist nanocoatings compared to controls. Conditioned medium collected from keratinocytes cultured on the IL-23R antagonist nanocoatings polarized macrophages toward a M2-like phenotype, based on increased CD163 and CD206 expression and reduced iNOS expression, compared to controls.Our results support development of IL-23R noncompetitive antagonist nanocoatings to reduce the pro-inflammatory IL-23/17A pathway and augment macrophage polarization toward a pro-regenerative phenotype. Immunomodulatory implant surface engineering may promote soft tissue healing and thereby reduce rates of peri-implantitis.Copyright © 2023 Elsevier Inc. All rights reserved

Development of standard protocols for biofilm-biomaterial interface testing

The oral biofilm is associated with the most common oral diseases such as caries, periodontitis, and peri-implantitis. It is also linked to failures of dental treatment approaches (eg, direct or indirect restorations because of adjacent caries). Therefore, the development of materials with antibacterial properties is desirable. However, the design of meaningful tests to confirm such properties faces severe problems because of the complexity of the interaction of materials with the oral biofilm. Furthermore, owing to practical reasons, such tests need to be performed in vitro. In contrast, there is a need for predictive data that are comparable between different laboratories. Therefore, standardization of such tests has been advocated. The International Organization for Standardization (ISO) with its Technical Committee 106–Dentistry maybetherelevant platform forthis purpose. A standard (ISO 3990) is being developed for testing the antibacterial properties of dental restorative materials. This standard defines basic requirements for sample preparation, selection of bacterial strains, test methods and assessment, and reporting of results. It is considered to be the first step, and regular revisions are planned as new scientific evidence emerges. The support of the scientific communities providing multidisciplinary input is needed. Key Words. Antibacterial; dental material; standardization; ISO standards; microbiology

Chitosan-recombinamer layer-by-layer coatings for multifunctional implants

Producción CientíficaThe main clinical problems for dental implants are (1) formation of biofilm around the implant—a condition known as peri-implantitis and (2) inadequate bone formation around the implant—lack of osseointegration. Therefore, developing an implant to overcome these problems is of significant interest to the dental community. Chitosan has been reported to have good biocompatibility and anti-bacterial activity. An osseo-inductive recombinant elastin-like biopolymer (P-HAP), that contains a peptide derived from the protein statherin, has been reported to induce biomineralization and osteoblast differentiation. In this study, chitosan/P-HAP bi-layers were built on a titanium surface using a layer-by-layer (LbL) assembly technique. The difference in the water contact angle between consecutive layers, the representative peaks in diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), X-ray photoelectron spectroscopy (XPS), and the changes in the topography between surfaces with a different number of bi-layers observed using atomic force microscopy (AFM), all indicated the successful establishment of chitosan/P-HAP LbL assembly on the titanium surface. The LbL-modified surfaces showed increased biomineralization, an appropriate mouse pre-osteoblastic cell response, and significant anti-bacterial activity against Streptococcus gordonii, a primary colonizer of tissues in the oral environmentMinisterio de Economía, Industria y Competitividad (Project MAT2013-42473-R and MAT2015-68901R)Junta de Castilla y León (programa de apoyo a proyectos de investigación – Ref. VA244U13, VA313U14 and VA015U16

Biomimetic Mineralization of Recombinamer-Based Hydrogels toward Controlled Morphologies and High Mineral Density

Producción CientíficaThe use of insoluble organic matrices as a structural template for the bottom-up fabrication of organic−inorganic nanocomposites is a powerful way to build a variety of advanced materials with defined and controlled morphologies and superior mechanical properties. Calcium phosphate mineralization in polymeric hydrogels is receiving significant attention in terms of obtaining biomimetic hierarchical structures with unique mechanical properties and understanding the mechanisms of the biomineralization process. However, integration of organic matrices with hydroxyapatite nanocrystals, different in morphology and composition, has not been well-achieved yet at nanoscale. In this study, we synthesized thermoresponsive hydrogels, composed of elastin-like recombinamers (ELRs), to template mineralization of hydroxyapatite nanocrystals using a biomimetic polymer-induced liquid-precursor (PILP) mineralization process. Different from conventional mineralization where minerals were deposited on the surface of organic matrices, they were infiltrated into the frameworks of ELR matrices, preserving their microporous structure. After 14 days of mineralization, an average of 78 μm mineralization depth was achieved. Mineral density up to 1.9 g/cm3 was found after 28 days of mineralization, which is comparable to natural bone and dentin. In the dry state, the elastic modulus and hardness of the mineralized hydrogels were 20.3 ± 1.7 and 0.93 ± 0.07 GPa, respectively. After hydration, they were reduced to 4.50 ± 0.55 and 0.10 ± 0.03 GPa, respectively. These values were lower but still on the same order of magnitude as those of natural hard tissues. The results indicated that inorganic−organic hybrid biomaterials with controlled morphologies can be achieved using organic templates of ELRs. Notably, the chemical and physical properties of ELRs can be tuned, which might help elucidate the mechanisms by which living organisms regulate the mineralization process.Junta de Castilla y León (programa de apoyo a proyectos de investigación – Ref. VA244U13

Automatic under-frequency load shedding mal-operation in power systems with high wind power penetration

[EN] Countries with a limited interconnection capacity suffer substantial frequency variations after large incidents so they use automatic under-frequency load shedding schemes to arrest the frequency decay. Some of these countries such as Portugal, Spain and Ireland also have very high wind penetrations. This can cause additional frequency excursions due to generation time variability but also to the fact that variable speed wind turbines do not add directly their inertia to the power system. Thus several transmission system operators have announced new grid codes requiring wind turbines to provide frequency response. In some scenarios, however, wind energy support may be detrimental to frequency control because it generates an extra energy that reduces decay and derivative but that cannot be maintained over time. These lower values of frequency decay and derivative are currently expected after a reduced incident or when conventional generation, which can maintain the extra generation, provides frequency support, so lead to low or no load shedding. This paper has studied, in particular, the effect of wind generation emulating inertia. A reduction of frequency derivative is achieved, which looks positive at first, but in some cases leads to initial smaller load shedding than the incident requires. A reduced frequency derivative triggers less under-frequency relays as if there were a significant amount of conventional generation that is online. However, this generation has been substituted by wind generation emulating inertia, and as it can maintain extra generation over time, the frequency continues to decay until the shedding of the next load step. As a result there is an excessive frequency deviation and an incorrect load shedding for the magnitude of the initial disturbance. In order to prevent this problem, automatic under frequency load shedding settings may need readjustment when a large amount of wind generation provides frequency support. (C) 2017 International Association for Mathematics and Computers in Simulation (IMACS). Published by Elsevier B.V. All rights reserved.The present work was supported by the Spanish Ministry of Economy and Competitivity and European Union FEDER funds under grant DPI2014-53245-R and by the Universitat Jaume I through project number P1 . 1B2015-42. Part of this work was done by Nestor Aparicio during a visit to University College Dublin, which was supported by the Spanish Ministry of Education, Culture and Sport under grant Jose Castillejo CAS14/00384.Aparicio Marín, N.; Añó Villalba, SC.; Belenguer Balaguer, E.; Blasco-Gimenez, R. (2018). Automatic under-frequency load shedding mal-operation in power systems with high wind power penetration. Mathematics and Computers in Simulation. 146:200-209. https://doi.org/10.1016/j.matcom.2016.12.006S20020914

Recombinant AMP/Polypeptide Self-Assembled Monolayers with Synergistic Antimicrobial Properties for Bacterial Strains of Medical Relevance

Producción CientíficaNosocomial infections are one of the most frequent causes of indwelling biomedical device failure. In this regard, the use of antibiofilm nanocoatings based on antimicrobial peptides (AMPs) is a promising alternative to prevent multiresistant biofilm infections. However, the limitations of chemical production impede the large-scale development of advanced antimicrobial materials that improve the properties of AMPs. Herein, we present a multifunctional modular design for the recombinant coproduction of self-assembled monolayers (SAMs) based on AMPs and elastin-like recombinamers (ELRs), which combine the antimicrobial properties of a designer AMP, GL13K, and low-fouling activity of an ELR in a synergistic manner. The inclusion of a grafting domain intended for oriented tethering onto surfaces allowed the recombinant polymers to be covalently immobilized onto model gold surfaces. The antibiofilm properties against two of the bacterial strains most frequently responsible for indwelling medical device-associated infections, namely Staphylococcus epidermidis and Staphylococcus aureus, were then evaluated. GL13K peptide was found to provide antibiofilm properties to the surface, with these being synergistically enhanced by the antifouling effect of the ELR. This new design offers a promising tool for the development of advanced AMP-based nanocoatings for medical devices with powerful and enhanced features.Comisión Europea (project NMP-2014-646075)Ministerio de Economía, Industria y Competitividad (project PCIN-2015-010 / MAT2015-68901-R / MAT2016-78903-R)Junta de Castilla y León (project VA317P18

Comportamiento a la corrosión de implantes de titanio granallados

Se ha demostrado que realizar el tratamiento de granallado sobre implantes mejora la fijación de los mismos, debido a la rugosidad superficial que adquieren. Variando el tamaño de las partículas de proyección del granallado, se varía el valor de rugosidad. Por otra parte, siempre quedan partículas adheridas en la superficie del implante, lo cual puede tener influencia en distintas propiedades, entre las cuales cabe destacar su resistencia a la corrosión. Este estudio determina el comportamiento a la corrosión, mediante mediciones potenciodinámicas, del titanio comercialmente puro granallado con diferentes materiales de partículas de proyección (Al2O3 y SiC) y con diferentes tamaños de partícula (200, 600 y 900 mm). La rugosidad no tuvo influencia significativa en los resultados de corrosión, pero sí la tuvo la naturaleza de las partículas de proyección. Así, los discos tratados con SiC, debido a la oxidación de estas partículas adheridas en la superficie del titanio, presentaron densidades de corriente un orden de magnitud mayores que los discos tratados con Al2O3, lo que les confiere peores características de resistencia a la corrosión. No obstante, todos los discos presentaron un adecuado comportamiento frente a la corrosión.Peer Reviewe