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Item Synthesis and Characterization of Chloroaluminium Phthalocyanine Incorporated in PLA/Gelatin Bionanocomposites for Photodynamic TherapyCarvalho, Janicy Arantes; Ambrosio, Jéssica Aparecida Ribeiro; Marcolino, Luciana Maria Cortez; Simioni, Andreza Ribeiro; Ribeirão PretoDevelopment delivery systems, such as nanoparticles, represent a growing area in biomedical research. Nanoparticles were prepared using a combination of two methods, the desolvation / emulsion technique using poly (lactic acid) (PLA) and gelatin (nPLA-G) forming a bionanocomposites which are hybrid materials composed of a biodegradable material and a nanodimensional material. Chloroaluminum phthalocyanine (ClAlPc) was chose as photosensitizer (PS). ClAlPc is a second generation of photosensitizer used in photodynamic therapy (PDT). ClAlPc loaded nanoparticles (nPLA-G/ClAlPc) were prepared using the same process. The PLA-gelatin nanoparticles were successfully prepared by the combination of desolvation / emulsion technique. The nanoparticles presented uniform size and spherical shape. The mean nanoparticle size was 655.1 ± 202.8 nm for nPLA-G (PDI = 0.163) and 772.2 ± 239.4 nm for nPLA-G/ClAlPc (PDI = 0.134). The zeta potential for nPLA-G and nPLA-G/ClAlPc revealed an average value of + 10.0 ± 3.15 mV and + 15.9 ± 3.29 mV, respectively. ClAlPc loaded nanoparticles maintain its photophysical behavior after encapsulation. The encapsulation efficiency was of 90.89 ± 7.55%. All the physical–chemical and photophysical measurements performed allow us to reinforce the idea of a new nanoparticle, composed of PLA and gelatin, as drug delivery systems for PDT.Item ATR-FTIR and Genetic Polymorphism Analysis: Potential Tools in the Diagnosis and Identification of Predisposition to DM2Silva, Ivan Saraiva; Diniz, Renata V.; Oliveira, Maria do Socorro Pombo de; Lima, Luciana de; Sibelino, Sônia Khouri; Raniero, Leandro José; Canevari, Renata de Azevedo; Ribeirão PretoType 2 Diabetes Mellitus (DM2) is a multifactorial disorder characterized by insulin resistance or reduced hormonal production, commonly diagnosed in adults. The pre-diabetes stage, reversible if treated, can progress to DM2, leading to serious complications such as vision loss and cardiovascular disease. Currently, conventional diagnostic methods, such as glucose measurement and glycated hemoglobin analysis, require fasting which can be uncomfortable for patients; thus, infrared spectroscopy analysis demonstrates great potential as a tool to facilitate disease diagnosis. Genetic polymorphism analysis, although expensive and time-consuming, offers early insights into predisposition to DM2, becoming an important technique for predicting susceptibility to disease development. Studies on the SNP -866G/A of the UCP2 gene, associated with DM2, indicate the polymorphism as one of the factors that may facilitate disease development. This study evaluates the feasibility of FTIR spectroscopy as an alternative method for DM2 and pre-diabetes diagnosis by analyzing serum collected from 17 patients (9 with elevated glucose and HbA1c levels and 8 controls). The presence of the -866G/A polymorphism in the UCP2 gene was evaluated using the PCR-RFLP technique for group stratification. The ATR-FTIR technique proved effective in separating patients with glucose alterations from those with normal glycemic patterns; however, further studies are needed to correlate the -866G/A polymorphism with DM2 predisposition.Item Assessment of the Effect of Antimicrobial Photodynamic Therapy on Multidrug- Resistant Acinetobacter baumannii Strain Biofilm - an in Vitro StudyBrambilla, Isabelle de Paula Ribeiro; Marcolino, Luciana Maria Cortez; Pinto, Juliana Guerra; Ferreira-Strixino, Juliana; Ribeirão PretoAcinetobacter baumannii is an opportunistic bacterium associated with numerous cases of hospital-acquired infections and possesses, among other characteristics that make it virulent, resistance to various antibiotics. The increasing resistance of microorganisms has led to the search for alternative methods, such as Antimicrobial Photodynamic Therapy (aPDT), inducing bacterial cell death. This study aimed to evaluate the antimicrobial action of PDT with curcumin on A. baumannii biofilm in vitro. A. baumannii biofilm was treated with curcumin for 20 min and then irradiated with a 450 nm LED (Light Emitting Diode) at 93.9 mW/cm2, with a fluence of 42.72 J/cm2. Subsequently, tests were conducted using Confocal Microscopy with the Resazurin dye, Colony-Forming Units (CFU) counting, and Scanning Electron Microscopy (SEM) imaging. Cell viability tests were also conducted using the L929 fibroblast cell line. The results show that although PDT with curcumin did not reduce bacterial growth, it caused alterations in cell metabolism. Additionally, confocal microscopy showed that curcumin was internalized by A. baumannii at the concentrations used. SEM demonstrated structural changes, although no bacterial reduction in the biofilm was observed. The parameters used in biofilm treatment were cytotoxic to L929 cells. This study showed that PDT with curcumin induces fragility in A. baumannii cells, suggesting that a possible combination with antibiotics may lead to the elimination of bacterial infection.Item Biomechanical analysis of experimental spinal cord injury treated with photobiomodulationBeloni, Luís Filipe Karatanasov; Lima, Leonardo Borges; Neves, Marcele Florencio; Lima, Mario Oliveira; Lima, Fernanda Pupio Silva; Lopes-Martins, Rodrigo Alvaro Brandão; Sant’Anna, Luciana Barros; Arisawa, Emilia Angela Lo Schiavo; Ribeirão PretoSpinal cord injury (SCI) is a terrible condition that can affect humans. It is characterized by the interruption of the neurological signal, affecting sensorimotor functions, with social and psychological losses and high treatment costs. There is still no cure for SCI, encouraging research for new therapeu- tic approaches. In this context, Photobiomodulation (PBM) has shown promising results regarding the inflammatory, pain, im- munological, and antifibrotic response, favoring the healing process in several tissues. Objective: To evaluate the motor function in rats treated with PBM after SCI by hemisection us- ing quantitative movement analysis techniques. Methodology: 15 male rats were distributed in three groups: Control (C), In- jury (I), and Photobiomodulation (PBM, continuous wave, 780nm). Groups I and PBM were submitted to SCI between T9/T10, by hemisection of the spinal cord. The Sciatic Func- tional Index (SFI) and average speed were evaluated by kine- matic analysis of the gait of these animals for 28 days, with 7- day intervals. Results: The PBM group showed a significant in- crease in the average speed at 7, 14, and 21 experimental day times evaluated and improvement in SFI. Conclusion: The kin- ematic and thesciatic functional index analysis are reliable eval- uating tools for motor function in rats treated with PBM after SCI, showing increased gait speed and range of motion im- provement on the 14th and 21st-day post-SCI.Item Enhanced Antimicrobial Strategy against Staphylococcus aureus: Synergistic Action of Plasma-Activated Water and Silver NanoparticlesSouza, Sophia Franzoni Azevedo; Costa, Maricilia Silva; Castilho, Maiara Lima; ParisThe rising prevalence of multidrug-resistant infections, particularly in chronic wounds of diabetic patients, necessitates the development of innovative antimicrobial approaches [1]. Staphylococcus aureus, a leading opportunistic pathogen in such cases, poses a significant challenge due to its antibiotic resistance and biofilm-forming capability [2]. This study investigates the combined antimicrobial efficacy of plasma-activated water (PAW), produced via a gliding arc plasma system, and silver nanoparticles (AgNPs), synthesized through a bottom-up chemical reduction method [3]. The primary objective was to determine whether the synergistic interaction between PAW-derived reactive species and AgNPs’ broad-spectrum antimicrobial properties could enhance bactericidal activity against S. aureus. Reactive species in PAW, including hydrogen peroxide (H₂O₂) and nitrite (NO₂⁻), were quantified using spectrophotometric colorimetric assays. Absorbance measurements, obtained with a BioTek Synergy HT microplate reader at 410 nm (H₂O₂) and 543 nm (NO₂⁻), demonstrated high linearity in standard calibration curves (R² > 0.99). Results revealed time-dependent variations in reactive species: H₂O₂ peaked at 15 minutes’ post-activation, while nitrite concentrations reached maximum levels at 2 hours before gradually declining. These dynamics underscore the importance of precise timing for PAW’s clinical application. Antimicrobial testing employed standardized S. aureus inoculum, evaluating individual and combined treatments of PAW and AgNPs over 15- and 60-minute incubation periods. Bare AgNPs achieved complete bacterial inhibition using 2,6x1011 particles/mL, while lower concentrations showed a number of particles-dependent reduction in viability. PAW alone exhibited time-dependent antimicrobial effects, with longer exposure (60 minutes) resulting in significantly greater bacterial inactivation compared to the 15-minute treatment. However, when PAW was combined with AgNPs, no significant difference was observed between the 15- and 60-minute treatment durations. Both conditions achieved similar inhibition of bacterial growth using low concentrations of AgNPs. These findings indicate that the synergistic antimicrobial effect of the PAW–AgNPs combination may compensate for shorter exposure times, enhancing efficacy even under reduced treatment durations. This synergistic effect is likely due to the combined mechanisms of oxidative stress induced by PAW’s reactive species and bactericidal action by AgNPs. In conclusion, the integration of PAW and AgNPs reduced S. aureus viability through complementary antimicrobial mechanisms. These results suggest a potential approach to support existing treatments for resistant infections, including those associated with chronic wounds. The combination of cold plasma and nanomaterials could pave the way for innovative antimicrobial solutions, offering enhanced support to conventional therapies in complex infection scenarios.Item Additive Manufacturing in Personalized Medicine: Enhancing the Development of Custom Biomodels Through 3D Scanning and Digital RiggingBasso, Beatriz Fernanda de Oliveira; Ortega, Leticia Martins Ramos; Ganga, Thabata; Rorato, Eduardo Keller; Lima, Mário Oliveira; Neves, Marcele Florêncio; Kunkel, Maria Elizete; Ribeirão Preto3D scanning technology and additive manufacturing techniques combined offer an innovative approach to creating personalized biomodels for medical applications. Biomodels, defined as mathematical, computational, or physical representations of a biological system, are increasingly being integrated into the design of medical devices, improving personalized medicine. Given its applications and growing interest, there is still a need to understand the challenges in manufacturing biomodels and a comprehensive methodology in the process of developing these 3D-scanned printed models. The objective of this paper is to investigate a fundamental methodology for developing biomodels to address complex clinical challenges, thereby enhancing precision and efficacy in medical practice. The methodology comprises a five-step process integrating 3D scanning, Computer Aided Design (CAD), Computer Aided Manufacturing (CAM), Computer Aided Engineering (CAE), and the “Rigging” technique. The results highlight the problems and promising clinical applications for the discussed methodology, notably in the manufacture of biomodels and the simulation of pathological structures. The spastic hand, proximal radius fracture, and healthy calf models demonstrated suitability for real-world application and high fidelity to their original anatomical form.Item 3D bioprinted human iPSC-derived neural progenitor cells as a novel platform for studying neurogenic niche(AIP Publishing) Machado, Lucas Simões; Ferreira, Paula Scanavez; Pires, Marina Rodrigues; Bim, Larissa Valdemarin; Oliveira, Natália Heloísa de; Salles, Geisa Rodrigues; Ferreira, Natalia Dall'Agnol; Cruz, Elisa Marozzi; Porcionatto, Marimelia AparecidaAnimal models, especially rodents, used to study neurodevelopment have significantly advanced our comprehension of cellular and molecular mechanisms. Nevertheless, differences in species-specific structures, gestation periods, and interneuronal connections limit animal models’ ability to represent human neurodevelopment accurately. The unique characteristics of primate neural progenitor cells (NPCs) enable cortex expansion with gyrus formation, which does not occur in lissencephalic animals, like rodents. Therefore, there is a need for novel in vitro models using human cells that recapitulate the complexity of human brain development. Along with organoids, 3D bioprinting offers a platform for creating more complex in vitro models. We developed, extensively characterized, and successfully used a GeltrexTM/GelMA hydrogel blend to bioprint human induced pluripotent stem cells-derived NPCs (hNPCs). We show that 3D bioprinted hNPCs can selforganize, revealing key features of a neurogenic niche, including proliferation, differentiation, and migration, remaining viable for over 110 days. Within the first 20 days, bioprinted constructs showed the formation of positive cell clusters for the neurogenic niche cell markers FABP7, NESTIN, and GFAP. Clusters were interconnected by process bundles supporting cell migration. The cells proliferated within the clusters, and over time, NPCs originated TUBB3þ neurons with long axonal tracts, prominent around the clusters. We propose this as a 4D model to study neurogenic niches’ key cellular and molecular features in a 3D bioprinted scaffold, adding time as the fourth dimension. Neuronal maturation in this dynamic model recapitulates key neurogenic niche properties, making it suitable for neurodevelopmental disease modeling and drug screening.Item The arrangement of dual-species biofilms of Candida albicans and Issatchenkia orientalis can be modified by the medium: effect of Voriconazole(Taylor & Francis) Passos, Juliene Cristina da Silva; Rodrigues, Ana Beatriz Furtado; Silva, Carlos Alberto; Costa, Maricilia SilvaBoth Candida albicans and Issatchenkia orientalis have been isolated from different types of infections over the years. They have the ability to form communities of microorganisms known as biofilms. It has been demonstrated that the medium employed in studies may affect the biofilm development. The aim of this study was to investigate the arrangement of dual-species biofilms of C. albicans and I. orientalis cultivated on either RPMI-1640 or Sabouraud Dextrose Broth (SDB), as well as the inhibitory effect of Voriconazole (VRC). For the experiments performed, ATCC strains were used, and yeast-mixed suspensions were inoculated in 96-well plates with either RPMI-1640 or SDB, in the presence or absence of VRC. The results were observed by counting the number of CFU obtained from scraping off the biofilms produced and plating the content on CHROMagar Candida medium. It was observed that for all conditions tested the medium chosen affected the arrangement of dual-species biofilms: when RPMI-1640 was used, there was a prevalence of C. albicans, while the opposite was noted when SDB was used. It could be suggested that the medium and environment could regulate interactions between both yeast species, including the response to different antifungal drugs.Item From the gut to the heart: probiotic therapy with Saccharomyces boulardii and its potential role on diabetic cardiomyopathy in a murine model(Taylor & Francis) Brandão, Ana Beatriz Pereira; Albuquerque, Raquel Cristina Melo Ferreira de; Abreu, Isabel Cristina Mallosto Emerich de; Ferreira, Fabiana Gomes; Santos, Leticia Barssotti dos; Jensen, Leonardo; Souza, Leandro Eziquiel de; Ferreira, Sarah Gomes; Souza, Lívia Bruni de; Arisawa, Emilia Angela Lo Schiavo; Sant′Anna, Luciana Barros; Higa, Elisa Mieko Suemitsu; Casali, Adenauer Girardi; Aimbire, Flávio; Irigoyen, Maria Claudia Costa; Casali, Karina Rabello; Cunha, Tatiana SousaBackground: We investigated whether the probiotic yeast Saccharomyces boulardii confers cardiometabolic protection and prevents diabetic cardiomyopathy by modulating inflammation, cardiac remodelling, cardiovascular function, and autonomic regulation. Methods: Male C57BL/6 mice were allocated into four groups: Control (C), Diabetes (DM), Control+Saccharomyces boulardii (CSb), and Diabetes+Saccharomyces boulardii (DMSb). Diabetes was induced with intraperitoneal streptozotocin (STZ), and treatments (sterile water or Saccharomyces boulardii) were administered orally for 8weeks. Blood glucose, cytokines, and nitric oxide levels were measured, along with cardiac function via echocardiography and direct blood pressure recordings. Results: Saccharomyces boulardii reduced blood glucose and increased cardiac IL-10 in diabetic mice, restoring nitric oxide levels. These effects were associated to reduced collagen deposition, preventing vascular damage and ventricular fibrosis, and were accompanied by improved systolic/ diastolic function and autonomic control. Conclusion: Saccharomyces boulardii improved cardiac structure, function, and autonomic control in diabetic mice, supporting its potential as adjunct therapy for diabetic cardiomyopathy.Item Targeted optimization of single-chain variable fragment (scFv) expression in E. coli using a design-of-experiment approach(Elsevier) Guimaraes, Marcela; Carvalho, Rafaela Vieira; Luz, Daniela; Guilherme, Ariela Pedro Bom; Campos, Claudia Barbosa Ladeira de; Piazza, Roxane Maria Fontes; Pradella, José Geraldo da CruzInfections by Shiga toxin-producing Escherichia coli (STEC) pose a serious global health risk, potentially leading to hemolytic uremic syndrome (HUS), a life-threatening condition. This study aimed to establish an efficient expression system to produce an anti-Stx2 single-chain variable fragment (scFv) using three E. coli strains: BL21 (DE3), BL21(DE3) pLysS, and ArcticExpress (DE3). Initial tests showed that BL21(DE3) pLysS and ArcticExpress (DE3) produced 0.3–0.4 mg scFv/L using defined media and induction with 0.1 mM IPTG or 10 g/L lactose. A Plackett-Burman design was then used to optimize the concentrations of 2xYT medium, IPTG, and lactose during induction. Under these conditions, ArcticExpress (DE3) reached 19 mg/L of scFv when induced with 1.0 mM IPTG and 2.5 g/L 2xYT. In contrast, BL21(DE3) pLysS yielded 34 mg/L with 1.0 mM IPTG, 0.4 g/L 2xYT, and 0.2 g/L lactose, while IPTG alone resulted in 26 mg/L. The protein yield (Yp/x) also increased from 11.9 to 15.5 mg scFv/g cell. Thus, E. coli BL21(DE3) pLysS showed the highest potential for producing anti-Stx2 scFv, particularly under optimized induction conditions involving IPTG, 2xYT, and lactose. These findings highlight the potential of recombinant E. coli BL21(DE3) pLysS as a robust platform for the scalable production of functional anti-Stx2 scFv fragments. By enabling higher yields under optimized induction strategies, this approach provides an important basis for the development of low-cost diagnostic tools targeting STEC infections, which remain a critical global health challenge.Item Simultaneous occurrence of midlatitude plasma bubbles and LSTIDs during the 10 October 2024 geomagnetic storm(Elsevier) Picanço, Giorgio Arlan da Silva; Fagundes, Paulo Roberto; Moro, Juliano; Nogueira, Paulo Alexandre Bronzato; Muella, Marcio Tadeu de Assis Honorato; Nardini, Clezio Marcos de; Resende, Laysa Cristina Araújo; Silva, Lígia Alves da; Laranja, Sophia Rodrigues; Anoruo, Chukwuma Moses; Agyei-Yeboah, Ebenezer; Souza, Ana Lucia Christovam deIn this study, we present a multi-instrumental analysis of the extreme geomagnetic storm of October 10, 2024, focusing on the inter- action between Equatorial Plasma Bubbles (EPBs) and Large-Scale Traveling Ionospheric Disturbances (LSTIDs) over the American sector. Using Rate of Total Electron Content Index (ROTI), Total Electron Content (TEC), and Detrended Total Electron Content (DTEC) maps derived from Global Navigation Satellite System (GNSS) data, we investigated the ionospheric response to the geomag- netic storm. This analysis was complemented by far-ultraviolet airglow observations from the Global-scale Observations of the Limb and Disk (GOLD) mission, in situ electron density profiles from the Swarm satellite constellation, and ground-based ionosonde measure- ments. These multi-instrumental datasets revealed a sequence of coupled processes responsible for the generation, expansion, and mor- phological deformation of EPBs, extending well beyond their typical equatorial domain. Prompt Penetration Electric Fields (PPEFs), which led to an enhanced pre-reversal enhancement (PRE) over western South America, drove a substantial uplift of the ionospheric F-region, exceeding 700 km, and favored the development of large-scale EPBs. These structures manifested as a reversed-C-shaped plasma depletion band, confirmed by both ROTI and GOLD observations. Simultaneously, DTEC maps and keograms revealed the equatorward propagation of storm-time LSTIDs. A pronounced spatial and temporal overlap between EPB and LSTID signatures was observed at midlatitudes, especially across the western South American longitudinal sector. This interaction appears to have mod- ulated the outer structure of the EPBs, leading to asymmetric deformation and enhanced latitudinal expansion. Finally, our findings highlight the role of multiscale coupling between high-latitude and equatorial processes during geomagnetic storms and emphasize the need for integrated observations to fully characterize the dynamics of storm-induced ionospheric disturbances.Item Retraction Note to: In vitro and in vivo evaluation of rotary-jet-spun poly(ɛ-caprolactone) with high loading of nano-hydroxyapatite(Springer) Andrade, Telmo M.; Mello, Daphne de Camargo Reis; Elias, Conceição de Maria Vaz; Abdala, Julia Marinzeck de Alcantara; Silva, Edmundo; Vasconcellos, Luana Marotta Reis de; Tim, Carla Roberta; Marciano, Fernanda Roberta; Lobo, Anderson OliveiraItem In vitro and in vivo evaluation of rotary-jet-spun poly(ɛ- caprolactone) with high loading of nano-hydroxyapatite(Springer) Andrade, Telmo M.; Mello, Daphne de Camargo Reis; Elias, Conceição de Maria Vaz; Abdala, Julia Marinzeck de Alcantara; Silva, Edmundo; Vasconcellos, Luana Marotta Reis de; Tim, Carla Roberta; Marciano, Fernanda Roberta; Lobo, Anderson OliveiraHerein, poly(ɛ-caprolactone) (PCL) mats with different amounts of nanohydroxyapatite (nHAp) were produced using rotary- jet spinning (RJS) and evaluated in vitro and in vivo. The mean fiber diameters of the PCL, PCL/nHAp (3%), PCL/nHAp (5%), and PCL/nHAp (20%) scaffolds were 1847±1039, 1817±1044, 1294±4274, and 845±248nm, respectively. Initially, all the scaffolds showed superhydrophobic behavior (contact angle around of 140oC), but decreased to 80° after 30 min. All the produced scaffolds were bioactive after soaking in simulated body fluid, especially PCL/nHAp (20%). The crystallinity of the PCL scaffolds decreased progressively from 46 to 21% after incorporation of 20% nHAp. In vitro and in vivo cytotoxicity were investigated, as well as the mats’ ability to reduce bacteria biofilm formation. In vitro cellular differentiation was evaluated by measuring alkaline phosphatase activity and mineralized nodule formation. Overall, we identified the total ideal amount of nHAp to incorporate in PCL mats, which did not show in vitro or in vivo cytotoxicity and promoted lamellar bone formation independently of the amounts of nHAp. The scaffolds with nHAp showed reduced bacterial proliferation. Alizarin red staining was higher in materials associated with nHAp than in those without nHAp. Overall, this study demonstrates that PCL with nHAp prepared by RJS merits further evaluation for orthopedic applicationsItem Physiological stress tolerance responses of the dung decomposer fungus Mucor circinelloides(Elsevier) Acheampong, Mavis Agyeiwaa; Pupin, Breno; Dias, Luciana Pereira Pinto; Santos, Maiara P.; Bernardes, Luana Oliveira; Pinheiro, Michele Souza; Ferreira, Paulo Cesar; Mikaelyan, Aram; Medina, Humberto Ramírez; Alder-Rangel, AleneThe Mucor circinelloides complex, a group of dung-dwelling fungi, is essential for nutrient cycling and organic matter breakdown. Despite their ecological significance, variations in germination rates and stress tolerance among isolates are not well understood. This study examined four isolates—Cicarelli, Doc Beaver, Papulsa, and Zaina—to assess their germination on various media, including potato dextrose agar (PDA), Czapek (CZA), Sabouraud dextrose agar (SDA), Emerson (EM), and minimal media (MM). The results showed notable differ- ences in germination rates, with CZA and EM promoting the fastest growth. The study also analysed spor- angiospore tolerance to UV-B radiation and high temperatures (47 ◦C), with findings highlighting the role of growth media in stress resistance. Sporangiospores from Cicarelli, Doc Beaver, and Papulsa grown on PDA exhibited greater UV-B tolerance than those on CZA. Conversely, sporangiospores produced on CZA were more heat tolerant than those on PDA. The 12-h survival curve for the Zaina isolate reinforced these findings, showing similar trends in UV-B resilience and heat tolerance. This research demonstrates how growth media influence the environmental stress responses in M. circinelloides, providing insights into the adaptive potential of these fungi.Item Cyto-genotoxicity and immunomodulation in fibroblasts exposed to calcium-aluminate cement with different radiopacifiers(Fundação Odontológica de Ribeirão Preto) Martorano, Antônio Secco; Messias, Nadyne Saab; Bighetti-Trevisan, Rayana Longo; Teixeira, Lucas Novaes; Oliveira, Ivone Regina de; Raucci-Neto, Walter; Oliveira, Paulo Tambasco de; Castro-Raucci, Larissa Moreira Spinola deCalcium aluminate cement (CAC) has been proposed as an alternative to mineral trioxide aggregate (MTA), but its biological behavior remains underexplored. This study evaluated the cytotoxicity, genotoxicity, and proinflammatory gene expression of fibroblasts exposed to CAC formulations containing zinc oxide or bismuth oxide with different proportions of calcium chloride, in comparison with MTA. Fibroblasts were cultured with CACz (25% ZnO + 2.8% CaCl₂), CACb (25% Bi₂O₃ + 2.8% CaCl₂), CACb+ (25% Bi₂O₃ + 10% CaCl₂), or MTA; non-exposed cells served as controls. Cytotoxicity was assessed by fluorescence microscopy and MTT viability assays at 24 h and 72 h, while genotoxicity was evaluated by the comet assay at 24 h. Gene expression of COL-1, TNF-α, IL-1β, IL-6, and MMP-9 was examined by RT-qPCR at 72 h. Data were analyzed using the Kruskal-Wallis test and the Student-Newman-Keuls post hoc test (α = 0.05). Cell viability did not differ among groups at 24 h, but was higher for CACb+ at 72 h (p < 0.05). DNA damage levels were similar across groups (p > 0.05). For gene expression, no significant differences were observed for COL-1, IL-1β, or TNF-α. All cements upregulated IL-6 compared with control (p < 0.05), with the highest levels in MTA and the lowest in CACb+. CACb+ increased MMP-9 expression (p < 0.05), while CACz showed the lowest levels. Within the limits of this study, CAC formulations with additives were not genotoxic or cytotoxic, and their biological behavior was composition-dependent, with CACb+ demonstrating the most favorable profile by enhancing fibroblast viability and extracellular matrix remodeling potential.Item Cost-effective production process of scFv antibody fragments against Shiga toxin 2 via recombinant E. coli(Elsevier) Guimarães, Marcela; Luz, Daniela; Augusto, Elisabeth de Fátima Pires; Vieira, Lucia; Costa, Maricilia Silva; Piazza, Roxane Maria Fontes; Pradella, José Geraldo da CruzShiga toxin (Stx)-producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are significant pathogens responsible for diarrhea, which can progress to hemorrhagic colitis and hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. Early diagnosis is crucial for effective clinical man- agement, as antibiotic treatment is not recommended for STEC infections. The present study aimed to establish a cost-effective biotechnological platform for cultivating recombinant E. coli to produce scFv antibody fragments against Stx2 for diagnostic applications. The method was first evaluated through shake flask experiments and subsequently scaled up to bench-scale bioreactors operated in both batch and fed-batch modes using defined culture media. Optimal production conditions were achieved by inducing recombinant E. coli pLys at 18 ◦C for 18 h with 0.1 mM IPTG, resulting in a yield of 3.0 to 4.0 mg scFv/g cell biomass. A fed-batch, high-cell-density procedure with E. coli pLysS achieved a maximum production up to 150 mg scFv/L. A preliminary economic assessment demonstrated the production potential at a value of around $250/g scFv. Economic analysis also highlights that the relative cost of capital investment becomes important as production processes intensify. Therefore, technical parameters such as productivity (scFv mass/bioreactor volume * time) and scFv concentration (mass scFv mass/bioreactor volume) should be prioritized to maximize their values. Similarly, optimization of the recombinant E. coli microbial platform should be pursued to increase the Yp/x level.Item In vitro evaluation of red and near infrared LED photobiomodulation on L929 fibroblasts metabolic activity and morphology(Springer) Segismondi, Larissa Cavallieri; Soares, Luís Eduardo Silva; Soares, Cristina PachecoPurpose Fibroblasts, the main cells of connective tissue, are highly susceptible to oxidative stress caused by reactive oxygen species (ROS). Photobiomodulation (PBM) has emerged as a promising therapy capable of modulating biological tissues, enhancing cellular metabolic activity, and promoting the proliferation and survival of fibroblasts. In this study, we aimed to investigate the potential in vitro photoprotective effect of cellular photomodulation using 660 and 850 nm LEDs in L929 fibroblast cells treated with hydrogen peroxide as a model of oxidative stress. Methods Changes in cell viability were observed using the Alamar Blue colorimetric assay, and cell morphology was assessed by inverted microscopy. Mitochondria and nuclei were also labeled in living cells using fluorescence with TMRM and Hoechst, in addition to ROS detection with CellRox Green. Results Our results indicate that pretreatment with LED exerts a cytoprotective effect against oxidative stress, promoting an increase in mitochondrial activity, mitochondrial membrane potential, and a reduction in intracellular reactive oxygen spe- cies (ROS) generation while inducing improvements in the morphological characteristics of cells. Conclusion The findings from the present study indicate that Photobiomodulation (PBM) with LED contributes to maintain- ing cellular homeostasis and can help prevent and mitigate damage resulting from oxidative stress in fibroblasts.Item Antimicrobial Photodynamic Therapy with Polystyrene Nanoparticles, Encapsulated Porphyrin-Derivative and Iodine Generation against Candida AlbicansCalvi, Gabriela de Souza; Braga, Marilia Toledo; Cartaxo, Giulia Nicolle Jácome; Liška, Vojtěch; Mosinger, Jiří; Costa, Maricília Silva; ParisItem Influence of fluoride characteristics on tooth surface protection in an erosive condition: a multifaceted characterization approach(Wiley) Silva, Marcelo Henrique Pereira Marques da; Laurindo, Vinicius Silva; Soares, Luís Eduardo SilvaAbstract Objective: To evaluate the effect of fluoride consistency and composition to protect enamel and dentin against the dental erosion. Materials and Methods: Bovine enamel and dentin specimens were treated with arti- ficial saliva, neutral fluoride gel (NFG), acidulated phosphate fluoride gel (AFG), neu- tral fluoride foam (NFF), and acidulated phosphate fluoride foam. The samples were subjected to cycling. Micro energy-dispersive X-ray fluorescence spectrometry, sur- face roughness (Ra), contact angle (CA), and scanning electron microscopy (SEM) were performed. Composition, CA and Ra data were analyzed by ANOVA and multi- ple comparison test (p < 0.05). Results: The dentin protected had a significantly higher mineral content than in the control. Eroded unprotected enamel had higher Ra values than normal surfaces. Fluo- ride treatments increased the Ra in dentin samples. AFG increased the CA in enamel. Fluoride foams increased CA in dentin with reduced mineral loss. SEM analysis found a deposited layer on enamel treated with AFG and remnants of deposits on dentin treated with NFG and NFF. Conclusion: Regardless of the form of application, fluoride provided protection against erosion, however with different levels. Clinical Significance: Applying the adequate fluoride form is relevant since the for- mulations have different effects on both enamel and dentin.Item Transcranial photobiomodulation therapy associated with cardiorespiratory rehabilitation in spastic subjects(Springer Nature Link) Pinto, Ana Paula; Lemos, Sergio Luiz; Fagundes, Alessandra de Almeida; Neves, Marcele Florêncio das; Martins, Rodrigo Álvaro Brandão Lopes; Silva, Fernanda Pupio; Silva, Mário OliveiraThe objective of this study was to evaluate the effects of cardiorespiratory rehabilitation (CR) and transcranial photobiomodu- lation (tPBM) on exercise tolerance (ET), heart rate variability (HRV), and peripheral muscle activity in individuals with spasticity. Fifteen participants with spasticity were randomly assigned to two groups: the tPBM group (tPBMG) consisted of eight volunteers who underwent tPBM (on mode) and CR, while the control group (CG) consisted of seven volunteers who underwent simulated tPBM (off mode) and CR. The CR program included 12 weeks of treatment, twice a week for one hour, involving aerobic exercises and lower limb strengthening. For tPBM, a cluster with three lasers (λ = 680 nm, 808 nm), with a power of 100 mW/laser and energy of 36 J, applied to the F7, F8, and Fpz points. The following parameters were evalu- ated after 8 and 12 weeks: ET, HRV, and surface electromyography (EMG) of the rectus femoris muscle during orthostasis (ORT), isometric squatting (ISOM), and isotonic squatting (ISOT). Both groups showed a 40% increase in ET for the CG and a 30% increase for the tPBMG. The CG had more pronounced parasympathetic modulation alterations during post-exercise effort and recovery compared to the tPBMG. The EMG results showed that the tPBMG exhibited progressive improvement in muscle activity during ISOM and ISOT, as well as a decrease in the interlimb difference. In conclusion, both CR and tPBMG demonstrated improvements in ET. However, tPBMG specifically showed promising effects on HRV modulation and peripheral muscle electrical activity, providing additional benefits compared to CR alone.