Evaluation of perioperative pachymetry of thin corneas during corneal crosslinking using hypo-osmolar riboflavin and hydroxypropyl methylcellulose

ABSTRACT Purpose: This study aimed to analyze variations in intraoperative corneal thickness during corneal cross-linking in patients with keratoconus and to investigate its possible correlation with presurgical maximal keratometry (Kmax) and pachymetry. Methods: This was a prospective case series. We used a method similar to the Dresden protocol, with the application of hydroxypropyl methylcellulose 0.1% hypo-osmolar riboflavin in corneas between 330 and 400 µm after epithelium removal. Corneal thickness was measured using portable calipers before and immediately after epithelium removal, and 30 and 60 min after the procedure. Results: The 30 patients in this study were followed up for one year. A statistically significant difference was observed in pachymetry values during the intraoperative period (p<0.0001) and an increase of 3.05 µm (95%C1: 0.56-5.54) for each diopter was seen after epithelium removal (p0.019). We found an average Kmax difference of —2.12 D between men and women (p0.013). One year after treatment, there was a statistically significant reduction in pachymetry (p<0.0001) and Kmax (p0.0170) values. Conclusions: A significant increase in pachymetry measurements was seen during the procedure, and most patients showed a regression in Kmax and pachymetry values one year after surgery.

Tumor-Suppressive Cross-Linking of Anti-T. cruzi Antibodies in Acute Lymphoblastic Leukemia.

Parasites have been associated with possible anticancer activity, including Trypanosoma cruzi, which has been linked to inhibiting the growth of solid tumors. To better understand this antitumor effect, we investigated the association of anti-T. cruzi antibodies with B cells of the acute lymphoblastic leukemia (ALL) SUPB15 cell line. The antibodies were generated in rabbits. IgGs were purified by affinity chromatography. Two procedures (flow cytometry (CF) and Western blot(WB)) were employed to recognize anti-T. cruzi antibodies on SUPB15 cells. We also used CF to determine whether the anti-T. cruzi antibodies could suppress SUPB15 cells. The anti-T. cruzi antibodies recognized 35.5% of the surface antigens of SUPB15. The complement-dependent cytotoxicity (CDC) results demonstrate the cross-suppression of anti-T. cruzi antibodies on up to 8.4% of SUPB15 cells. For the WB analysis, a band at 100 kDa with high intensity was sequenced using mass spectrometry, identifying the protein as nucleolin. This protein may play a role in the antitumor effect on T. cruzi. The anti-T. cruzi antibodies represent promising polyclonal antibodies that have the effect of tumor-suppressive cross-linking on cancer cells, which should be further investigated.

Innovative Approach to Accelerate Wound Healing: Synthesis and Validation of Enzymatically Cross-Linked COL-rGO Biocomposite Hydrogels.

In this study, an innovative conductive hybrid biomaterial was synthetized using collagen (COL) and reduced graphene oxide (rGO) in order for it to be used as a wound dressing. The hydrogels were plasticized with glycerol and enzymatically cross-linked with horseradish peroxidase (HRP). A successful interaction among the components was demonstrated by FTIR, XRD, and XPS. It was demonstrated that increasing the rGO concentration led to higher conductivity and negative charge density values. Moreover, rGO also improved the stability of hydrogels, which was expressed by a reduction in the biodegradation rate. Furthermore, the hydrogel's stability against the enzymatic action of collagenase type I was also strengthened by both the enzymatic cross-linking and the polymerization of dopamine. However, their absorption capacity, reaching values of 215 g/g, indicates the high potential of the hydrogels to absorb fluids. The rise of these properties positively influenced the wound closure process, achieving an 84.5% in vitro closure rate after 48 h. These findings clearly demonstrate that these original composite biomaterials can be a viable choice for wound healing purposes.

Biodegradable Biocomposite of Starch Films Cross-Linked with Polyethylene Glycol Diglycidyl Ether and Reinforced by Microfibrillated Cellulose.

Biopolymers are biodegradable and renewable and can significantly reduce environmental impacts. For this reason, biocomposites based on a plasticized starch and cross-linker matrix and with a microfibrillated OCC cardboard cellulose reinforcement were developed. Biocomposites were prepared by suspension casting with varied amounts of microfibrillated cellulose: 0, 4, 8, and 12 wt%. Polyethylene glycol diglycidyl ether (PEGDE) was used as a cross-linking, water-soluble, and non-toxic agent. Microfibrillated cellulose (MFC) from OCC cardboard showed appropriate properties and potential for good performance as a reinforcement. In general, microfiber incorporation and matrix cross-linking increased crystallization, reduced water adsorption, and improved the physical and tensile properties of the plasticized starch. Biocomposites cross-linked with PEGDE and reinforced with 12 wt% MFC showed the best properties. The chemical and structural changes induced by the cross-linking of starch chains and MFC reinforcement were confirmed by FTIR, NMR, and XRD. Biodegradation higher than 80% was achieved for most biocomposites in 15 days of laboratory compost.

Corneal Collagen Crosslinking for Ectasia After Refractive Surgery: A Systematic Review and Meta-Analysis.

Introduction: Corneal ectasia leads to progressive irregular corneal curvature and reduced visual acuity. Objective: To assess the safety and effectiveness of corneal collagen cross-linking (CXL) for managing corneal ectasia resulting from refractive laser surgery (RSL). Methods: A systematic review and meta-analysis were realized according to PRISMA guidelines. We searched PubMed, EMBASE, Cochrane, and Web of Science databases for studies on CXL in patients with ectasia after RLS. The outcomes of interest included visual acuity, refractive outcomes, topographic parameters (Kmax, index surface variance (ISV), index of Vertical Asymmetry (IVA), keratoconus index (KI), central keratoconus index (CKI), index of height asymmetry (IHA), index of height decentration (IHD) and Rmin (minimum sagittal curvature)), central corneal thickness, endothelial cell count, and possible adverse events. Statistical analysis was performed using the R software (version 4.2.3, R Foundation for Statistical Computing, Vienna, Austria). Results: 15 studies encompassing 421 patients (512 eyes) were included. The mean age was 32.03 ± 4.4 years. The pooled results showed a stable uncorrected visual acuity post-CXL, with a significant improvement in corrected distance visual acuity (SMD = 0.09; 95% CI: -0.07 to 0.26). The spherical equivalent decreased significantly (SMD = -0.09; 95% CI: -0.35, -0.02). The topographic parameter Kmax decreased significantly (SMD = 0.15; 95% CI:0.01 to 0.28); however, the other parameters, ISV, IVA, KI, CKI, IHA, IHD, and Rmin, did not change significantly. Central corneal thickness decreased significantly (SMD = 0.24; 95% CI:0.07 to 0.41), and the endothelial cell count remained stable The complications were rare. Conclusion: CXL is a safe and effective technique for managing corneal ectasia after RLS.

The Controlled Release of Abscisic Acid (ABA) Utilizing Alginate-Chitosan Gel Blends: A Synergistic Approach for an Enhanced Small-Molecule Delivery Controller.

The integration of abscisic acid (ABA) into a chitosan-alginate gel blend unveils crucial insights into the formation and stability of these two substances. ABA, a key phytohormone in plant growth and stress responses, is strategically targeted for controlled release within these complexes. This study investigates the design and characterization of this novel controlled-release system, showcasing the potential of alginate-chitosan gel blends in ABA delivery. Computational methods, including molecular dynamics simulations, are employed to analyze the structural effects of microencapsulation, offering valuable insights into complex behavior under varying conditions. This paper focuses on the controlled release of ABA from these complexes, highlighting its strategic importance in drug delivery systems and beyond. This controlled release enables targeted and regulated ABA delivery, with far-reaching implications for pharmaceuticals, agriculture, and plant stress response studies. While acknowledging context dependency, the paper suggests that the liberation or controlled release of ABA holds promise in applications, urging further research and experimentation to validate its utility across diverse fields. Overall, this work significantly contributes to understanding the characteristics and potential applications of chitosan-alginate complexes, marking a noteworthy advancement in the field of controlled-release systems.

RawVegetable 2.0: Refining XL-MS Data Acquisition through Enhanced Quality Control.

We present RawVegetable 2.0, a software tailored for assessing mass spectrometry data quality and fine-tuned for cross-linking mass spectrometry (XL-MS) applications. Building upon the capabilities of its predecessor, RawVegetable 2.0 introduces four main modules, each providing distinct and new functionalities: 1) Pair Finder, which identifies ion doublets characteristic of cleavable cross-linking experiments; 2) Diagnostic Peak Finder, which locates potential reporter ions associated with a specific cross-linker; 3) Precursor Signal Ratio, which computes the ratio between precursor intensity and the total signal in an MS/MS scan; and 4) Xrea, which evaluates spectral quality by analyzing the heterogeneity of peak intensities within a spectrum. These modules collectively streamline the process of optimizing mass spectrometry data acquisition for both Proteomics and XL-MS experiments. RawVegetable 2.0, along with a comprehensive tutorial is freely accessible for academic use at: http://patternlabforproteomics.org/rawvegetable2.

Chemical analysis of n-propyl gallate used as pre-treatment for resin-dentin bond strength: In vitro study.

This study aimed to evaluate the effect of n-propyl gallate as pre-treatment for resin-dentin bond strength. The dentin pre-treatments evaluated included propyl gallate of concentrations 0.1% (w/v), 1.0% (w/v), and 10.0% (w/v), as well as glutaraldehyde 5.0% (v/v), and distilled water as a control treatment. Dentin specimens were prepared for Fourier Transformed Infrared Spectroscopy (FT-IR) (n = 3/pre-treatment). Pre-treatments were actively applied to dentin blocks before performing the adhesive procedure to composite resin. Microtensile bond strength to dentin (µTBS) (n = 8/pre-treatment) was determined after 24 h and 6 months of storage. Data were submitted to a two-way ANOVA, followed by Tukey's post hoc test. As for FT-IR, propyl gallate 1%-treated specimens presented higher water, carbonate, collagen, and amide absorbance rates compared to other tested groups, while specimens pre-treated with glutaraldehyde and distilled water presented similar absorbance curves. Regarding µTBS, all concentrations of propyl gallate resulted in statistically significant higher bond strength values than distilled water at 24 h. After 6 months of storage, propyl gallate 0.1% was the only group that maintained µTBS over time. Propyl gallate 0.1% might be a suitable dentinal pre-treatment due to being able to present chemical bonds with demineralized dentin and providing resin-dentin bond stability after 6 months of storage.

Development and characterization of films from Campomanesia xanthocarpa and commercial citrus pectins with different degrees of methyl-esterification.

In this study, pectins from commercial citrus and isolated from gabiroba (Campomanesia xanthocarpa) fruits, were obtained with different degrees of methyl-esterification (DM) and applied in the films. The DM ranged from 0 % to 62.5 % and the gradual de-esterification process was confirmed by mono-dimensional analysis (1H NMR). In order to investigate the influence of DM values in pectin film properties, PCP (DM: 62.5 %); PCP-5 (DM: 37.4 %); PCP-15 (DM: 19.1 %), and a fully de-esterified sample PCP-35 (DM: 0 %) were selected. The functional properties of the films clearly showed that the DM and cross-linking process are necessary to obtain a material with water resistance. Furthermore, pectin isolated from the fruits of gabiroba was purified (GW-Na, DM: 51.9 %) and partially de-esterified (GW-Na-5, DM: 37.1 %). These pectins were used, for the first time, in development of films and the physical and mechanical properties were compared with films made with PCP and PCP-5 samples. GW-Na and GW-Na-5 films presented suitable properties, with reduced solubility reduced (57.1 and 26.2 %), high degree of swelling (2.14 and 2.26), low flexibility (18.05 and 6.11 MPa), respectively. High strength and rigidity (99.36 and 1040.9 MPa), for both films (GW-Na and GW-Na-5) were demonstrated, similar to that obtained by analyzed citrus pectin.

Evaluation of unilateral corneal collagen cross-linking on fellow untreated eyes of patients with keratoconus / A avaliação da reticulação unilateral do colágeno corneano em olhos não tratados de pacientes com ceratocone

ABSTRACT Purpose: This study aimed to examine the effects of unilateral corneal collagen cross-linking treatment on visual acuity and the topographic findings of the fellow untreated eye of patients who had bilateral progressive keratoconus. Methods: Patients with progressive keratoconus who underwent cross-linking treatment were screened retrospectively. A total of 188 untreated eyes of 188 patients whose eyes were treated unilaterally with either standard or accelerated cross-linking and refused cross-linking procedure for the fellow eye were included. Visual acuity and topographic findings of the fellow untreated eyes were obtained preoperatively and postoperatively at the 1st, 3rd, 6th, 12th, 24th, 30th, and 36th months. Results: The change over time of variables examined was similar in the untreated eyes of patients who received standard and accelerated cross-linking methods (p>0.05). At the 12th month, 136 (95.8%) untreated eyes were stable according to progression criteria. Only 4 (8%) eyes were progressive at the 24th month. No progression was observed in any of the 16 patients with a 36-month follow up. Conclusions: The results showed that the fellow untreated eyes of patients with bilateral progressive keratoconus did not have significant progression rates after unilateral cross-linking treatment.

RESUMO Objetivo: Examinar os efeitos do tratamento de reticulação unilateral do colágeno corneano na acuidade visual e os achados topográficos em olhos não tratados de pacientes com ceratocone progressivo bilateral. Métodos: Foram rastreados retrospectivamente pacientes com ceratocone progressivo submetidos a tratamento de reticulação. Foram incluídos no estudo 188 olhos não tratados de 188 pacientes tratado unilateralmente com reticulação padrão ou acelerada e que recusaram o procedimento de reticulação no outro olho. A acuidade visual e os achados topográficos dos olhos não tratados foram obtidos no pré- e pós-operatório no 1º, 3º, 6º, 12º, 24º, 30º e 36º mês. Resultados: As alterações ao longo do tempo foram semelhantes para as variáveis examinadas nos olhos não tratados de pacientes tratados com métodos de reticulação padrão e acelerado (p>0,05). No 12º mês, 136 olhos não tratados (95,8%) estavam estáveis, de acordo com os critérios de progressão. Apenas quatro olhos (8%) mostraram progressão no 24º mês. Nenhuma progressão foi observada nos 16 pacientes que tiveram um acompanhamento de 36 meses. Conclusões: O estudo mostrou que os olhos não tratados de pacientes com ceratocone progressivo bilateral não apresentaram taxas de progressão significativas após o tratamento unilateral com reticulação.

Long-term follow-up of repeated corneal cross-linking for progressive keratoconus in young patients / Acompanhamento a longo prazo do retratamento com cross-linking corneano para ceratocone em progressão em pacientes jovens

ABSTRACT Objective: To evaluate the long-term safety and efficacy of repeated corneal cross-linking in eyes of children and adolescents with progressive keratoconus. Methods: This retrospective study included nine eyes of nine consecutive patients with progressive keratoconus who underwent repeated corneal cross-linking 3.9 (range of 1.6 to 5.6) years after a primary one. All patients were followed for a mean period of 9.11 (range of 6 to 11) years after first treatment and 5.44 (range of 4 to 9) years, after corneal cross-linking retreatment. Results: Nine eyes of nine patients (six male) with progressive keratoconus underwent primary corneal cross-linking from 2009 to 2011. Despite the stability achieved with the epi-off corneal cross-linking, keratoconus continued to progress after some time. Mean time to documented evidence of keratoconus progression after primary corneal cross-linking was 3.9 (range of 1.6 to 5.6) years. All eyes were retreated as soon as progression was noted. At the last follow-up visit, 5.44 (range of 4 to 9) years after repeated corneal cross-linking, there was a significant decrease of 2.02 D in mean maximum topographic K-value (p = 0.045) and 1.95D in mean topographic K-value (p = 0.007). Conclusion: Repeated corneal cross-linking seems to be a safe and effective procedure to halt keratoconus progression after a primary corneal cross-linking failure.

RESUMO Objetivo: Avaliar a segurança e a eficácia a longo prazo da repetição do cross-linking corneano em olhos de crianças e adolescentes com ceratocone progressivo. Métodos: Estudo retrospectivo que incluiu nove olhos de nove pacientes consecutivos com ceratocone progressivo, que foram submetidos a retratamento com cross-linking corneano de 3,9 (variação de 1,6 a 5,6) anos após tratamento primário com cross-linking corneano. Todos os pacientes foram acompanhados por um período médio de 9,11 (variação de 6 a 11) anos após o primeiro tratamento e 5,44 (variação de 4 a 9) anos após retratamento com cross-linking corneano. Resultados: Nove olhos de nove pacientes (seis homens) com progressão do ceratocone foram submetidos a cross-linking corneano primário de 2009 a 2011. Apesar da estabilidade alcançada com o epi-off cross-linking corneano, o ceratocone continuou a progredir após algum tempo. O tempo médio para evidência documentada de progressão do ceratocone após cross-linking corneano primário foi de 3,9 (intervalo de 1,6 a 5,6) anos. Todos os olhos foram retratados com cross-linking corneano, assim que a progressão foi observada. Na última consulta de acompanhamento, 5,44 (variação de 4 a 9) anos após a repetição do cross-linking corneano, houve diminuição significativa de 2,02 D na média da curvatura máxima (p = 0,045) e 1,95 D na média da curvatura média (p = 0,007). Conclusão: O retratamento com cross-linking corneano parece ser um procedimento seguro e eficaz para interromper a progressão do ceratocone após falha primária do cross-linking corneano.

In Situ Photo-crosslinkable Hyaluronic Acid/Gelatin Hydrogel for Local Nitric Oxide Delivery.

An increasing number of studies have shown that the local release of nitric oxide (NO) from hydrogels stimulates tissue regeneration by modulating cell proliferation, angiogenesis, and inflammation. The potential biomedical uses of NO-releasing hydrogels can be expanded by enabling their application in a fluid state, followed by controlled gelation triggered by an external factor. In this study, we engineered a hydrogel composed of methacrylated hyaluronic acid (HAGMA) and thiolated gelatin (GELSH) with the capacity for in situ photo-cross-linking, coupled with localized NO release. To ensure a gradual and sustained NO release, we charged the hydrogels with poly(l-lactic-co-glycolic acid) (PLGA) nanoparticles functionalized with S-nitrosoglutathione (GSNO), safeguarding SNO group integrity during photo-cross-linking. The formation of thiol-ene bonds via the reaction between GELSH's thiol groups and HAGMA's vinyl groups substantially accelerated gelation (by a factor of 6) and increased the elastic modulus of hydrated hydrogels (by 1.9-2.4 times). HAGMA/GELSH hydrogels consistently released NO over a 14 day duration, with the release of NO depending on the hydrogels' equilibrium swelling degree, determined by the GELSH-to-HAGMA ratio. Biocompatibility assessments confirmed the suitability of these hydrogels for biological applications as they display low cytotoxicity and stimulated fibroblast adhesion and proliferation. In conclusion, in situ photo-cross-linkable HAGMA/GELSH hydrogels, loaded with PLGA-GSNO nanoparticles, present a promising avenue for achieving localized and sustained NO delivery in tissue regeneration applications.

Chia Oil Microencapsulation Using Tannic Acid and Soy Protein Isolate as Wall Materials.

The use of proteins to produce oil-containing microcapsules has been previously analyzed; however, their chemical modification, in order to improve their performance as wall materials, is a strategy that has not been widely developed yet. This study aimed to analyze the chemical modification of the proteins through cross-linking reactions with tannic acid and to evaluate their performance as wall materials to the microencapsulation of oils rich in polyunsaturated fatty acids. The cross-linking reaction of isolated soy protein and tannic acid was carried out at pH 10-11 and 60 °C. Subsequently, emulsions were made with a high-speed homogenizer and microcapsules were obtained by spray drying. Microcapsules were characterized by particle size, morphology (SEM), total pore area and % porosity (mercury intrusion methodology), superficial properties (contact angle), and size distribution of oil droplets (by laser diffraction). Additionally, encapsulation efficiency was determined as a function of total and surface oil. Oil chemical stability and quality were studied by Rancimat, hydroperoxide values, and fatty acid profiles. In addition, a storage test was performed for 180 days, and released oil and polyphenols were determined by in vitro gastric digestion. Moreover, the fatty acid composition of the oil and the total polyphenol content and antioxidant capacity of polyphenols were analyzed. The results showed that spray-dried microcapsules had an encapsulation efficiency between 54 and 78%. The oxidative stability exhibited a positive correlation between the amount of polyphenols used and the induction time, with a maximum of 27 h. The storage assay showed that the peroxide value was lower for those cross-linked microcapsules concerning control after 180 days. After the storage time, the omega-3 content was reduced by 49% for soy protein samples, while cross-linked microcapsules maintained the initial concentration. The in-vitro digestion assay showed a decrease in the amount of oil released from the cross-linked microcapsules and an increase in the amount of polyphenols and a higher antioxidant capacity for all samples (for example, 238.10 mgGAE/g and 554.22 mg TE/g for undigested microcapsules with TA 40% versus 322.09 mgGAE/g and 663.61 mg TE/g for digested samples). The microcapsules showed a high degree of protection of the encapsulated oil, providing a high content of polyunsaturated fatty acids (PUFAS) and polyphenols even in prolonged storage times.

Nanocatalytic performance of pectinase immobilized over in situ prepared magnetic nanoparticles.

Immobilization of enzymes is one of the protein engineering methods used to improve their thermal and long-term stabilities. Immobilized pectinase has become an essential biocatalyst for optimization in the food processing industry. Herein, nanostructured magnetic nanoparticles were prepared in situ for use as supports to immobilize pectinase. The structural, morphological, optical and magnetic features and the chemical compositions of the nanoparticles were characterized. Nanoparticle agglomeration and low porosity were observed due to the synthetic conditions. These nanoparticles exhibited superparamagnetic behavior, which is desirable for biotechnological applications. The maximum retention rate for the enzyme was observed at pH 4.5 with a value of 1179.3 U/mgNP (units per milligram of nanoparticle), which was equivalent to a 65.6% efficiency. The free and immobilized pectinase were affected by the pH and temperature. The long-term instability caused 40% and 32% decreases in the specific activities of the free and immobilized pectinase, respectively. The effects of immobilization were analyzed with kinetic and thermodynamic studies. These results indicated a significant affinity for the substrate, a decreased reaction rate, and improved thermal stability of the immobilized pectinase. The reusability of the immobilized pectinase was preserved effectively during cycling, with only a 21.2% decrease in activity observed from the first to the last use. Therefore, alternative magnetic nanoparticles are presented for immobilizing and maintaining the thermostability of pectinase.

Development and Evaluation of Cross-Linked Alginate-Chitosan-Abscisic Acid Blend Gel.

Abscisic acid (ABA) has been proposed to play a significant role in the ripening of nonclimacteric fruit, stomatal opening, and response to abiotic stresses in plants, which can adversely affect crop growth and productivity. The biological effects of ABA are dependent on its concentration and signal transduction pathways. However, due to its susceptibility to the environment, it is essential to find a suitable biotechnological approach to coat ABA for its application. One promising approach is to utilize alginate and chitosan, two natural polysaccharides known for their strong affinity for water and their ability to act as coating agents. In this study, an alginate-chitosan blend was employed to develop an ABA cover. To achieve this, an alginate-chitosan-abscisic acid (ALG-CS-ABA) blend was prepared by forming ionic bonds or complexes with calcium ions, or through dual cross-linking. This was done by dripping a homogeneous solution of alginate-chitosan and ABA into a calcium chloride solution, resulting in the formation of the blend. By combining the unique properties of alginate, chitosan, and ABA, the resulting ALG-CS-ABA blend can potentially offer enhanced stability, controlled release, and improved protection of ABA. These characteristics make it a promising biotechnological approach for various applications, including the targeted delivery of ABA in agricultural practices or in the development of innovative plant-based products. Further evaluation and characterization of the ALG-CS-ABA blend will provide valuable insights into its potential applications in the fields of biomedicine, agriculture, and tissue engineering.

Chitosan, Gelatin, and Collagen Hydrogels for Bone Regeneration.

Hydrogels are versatile biomaterials characterized by three-dimensional, cross-linked, highly hydrated polymeric networks. These polymers exhibit a great variety of biochemical and biophysical properties, which allow for the diffusion of diverse molecules, such as drugs, active ingredients, growth factors, and nanoparticles. Meanwhile, these polymers can control chemical and molecular interactions at the cellular level. The polymeric network can be molded into different structures, imitating the structural characteristics of surrounding tissues and bone defects. Interestingly, the application of hydrogels in bone tissue engineering (BTE) has been gathering significant attention due to the beneficial bone improvement results that have been achieved. Moreover, essential clinical and osteoblastic fate-controlling advances have been achieved with the use of synthetic polymers in the production of hydrogels. However, current trends look towards fabricating hydrogels from biological precursors, such as biopolymers, due to the high biocompatibility, degradability, and mechanical control that can be regulated. Therefore, this review analyzes the concept of hydrogels and the characteristics of chitosan, collagen, and gelatin as excellent candidates for fabricating BTE scaffolds. The changes and opportunities brought on by these biopolymers in bone regeneration are discussed, considering the integration, synergy, and biocompatibility features.

Exploring the Interplay between Polyphenols and Lysyl Oxidase Enzymes for Maintaining Extracellular Matrix Homeostasis.

Collagen, the most abundant structural protein found in mammals, plays a vital role as a constituent of the extracellular matrix (ECM) that surrounds cells. Collagen fibrils are strengthened through the formation of covalent cross-links, which involve complex enzymatic and non-enzymatic reactions. Lysyl oxidase (LOX) is responsible for catalyzing the oxidative deamination of lysine and hydroxylysine residues, resulting in the production of aldehydes, allysine, and hydroxyallysine. These intermediates undergo spontaneous condensation reactions, leading to the formation of immature cross-links, which are the initial step in the development of mature covalent cross-links. Additionally, non-enzymatic glycation contributes to the formation of abnormal cross-linking in collagen fibrils. During glycation, specific lysine and arginine residues in collagen are modified by reducing sugars, leading to the creation of Advanced Glycation End-products (AGEs). These AGEs have been associated with changes in the mechanical properties of collagen fibers. Interestingly, various studies have reported that plant polyphenols possess amine oxidase-like activity and can act as potent inhibitors of protein glycation. This review article focuses on compiling the literature describing polyphenols with amine oxidase-like activity and antiglycation properties. Specifically, we explore the molecular mechanisms by which specific flavonoids impact or protect the normal collagen cross-linking process. Furthermore, we discuss how these dual activities can be harnessed to generate properly cross-linked collagen molecules, thereby promoting the stabilization of highly organized collagen fibrils.

Development of a Redox-Polymer-Based Electrochemical Glucose Biosensor Suitable for Integration in Microfluidic 3D Cell Culture Systems.

The inclusion of online, in situ biosensors in microfluidic cell cultures is important to monitor and characterize a physiologically mimicking environment. This work presents the performance of second-generation electrochemical enzymatic biosensors to detect glucose in cell culture media. Glutaraldehyde and ethylene glycol diglycidyl ether (EGDGE) were tested as cross-linkers to immobilize glucose oxidase and an osmium-modified redox polymer on the surface of carbon electrodes. Tests employing screen printed electrodes showed adequate performance in a Roswell Park Memorial Institute (RPMI-1640) media spiked with fetal bovine serum (FBS). Comparable first-generation sensors were shown to be heavily affected by complex biological media. This difference is explained in terms of the respective charge transfer mechanisms. Under the tested conditions, electron hopping between Os redox centers was less vulnerable than H2O2 diffusion to biofouling by the substances present in the cell culture matrix. By employing pencil leads as electrodes, the incorporation of these electrodes in a polydimethylsiloxane (PDMS) microfluidic channel was achieved simply and at a low cost. Under flow conditions, electrodes fabricated using EGDGE presented the best performance with a limit of detection of 0.5 mM, a linear range up to 10 mM, and a sensitivity of 4.69 µA mM-1 cm-2.

The Non-Canonical Iron-Responsive Element of IRE-tvcp12 Hairpin Structure at the 3'-UTR of Trichomonas vaginalis TvCP12 mRNA That Binds TvHSP70 and TvACTN-3 Can Regulate mRNA Stability and Amount of Protein.

Trichomonas vaginalis is one of the most common sexually transmitted parasites in humans. This protozoan has high iron requirements for growth, metabolism, and virulence. However, iron concentrations also differentially modulate T. vaginalis gene expression as in the genes encoding cysteine proteinases TvCP4 and TvCP12. Our goal was to identify the regulatory mechanism mediating the upregulation of tvcp12 under iron-restricted (IR) conditions. Here, we showed by RT-PCR, Western blot, and immunocytochemistry assays that IR conditions increase mRNA stability and amount of TvCP12. RNA electrophoretic mobility shift assay (REMSA), UV cross-linking, and competition assays demonstrated that a non-canonical iron-responsive element (IRE)-like structure at the 3'-untranslated region of the tvcp12 transcript (IRE-tvcp12) specifically binds to human iron regulatory proteins (IRPs) and to atypical RNA-binding cytoplasmic proteins from IR trichomonads, such as HSP70 and α-Actinin 3. These data were confirmed by REMSA supershift and Northwestern blot assays. Thus, our findings show that a positive gene expression regulation under IR conditions occurs at the posttranscriptional level possibly through RNA-protein interactions between atypical RNA-binding proteins and non-canonical IRE-like structures at the 3'-UTR of the transcript by a parallel mechanism to the mammalian IRE/IRP system that can be applied to other iron-regulated genes of T. vaginalis.

Dental Adhesives-Surface Modifications of Dentin Structure for Stable Bonding.

The latest advancements in dentin bonding have focused on strategies to impair degradation mechanisms in order to extend the longevity of bonded interfaces. Protease inhibitors can reduce collagen degradation within the hybrid layer (HL). Collagen cross-linkers allow better adhesive infiltration and also inhibit proteases activity. Particles added to adhesive can promote mineral precipitation within the HL, reducing nanoleakage and micropermeability, besides possible antimicrobial and enzymatic inhibition effects. Most of these approaches are still experimental, and aspects of the adhesive under the clinician's control are still determinant for the long-term stability of adhesive restorations.