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Bone tissue engineering is a promising alternative to repair wounds caused by cellular or physical accidents that humans face daily. In this sense, the search for new graphene oxide (GO) nanofillers related to their degree of oxidation is born as an alternative bioactive component in forming new scaffolds. In the present study, three different GOs were synthesized with varying degrees of oxidation and studied chemically and tissue-wise. The oxidation degree was determined through infrared (FTIR), X-ray diffraction (XRD), X-ray photoelectron (XPS), and Raman spectroscopy (RS). The morphology of the samples was analyzed using scanning electron microscopy (SEM). The oxygen content was deeply described using the deconvolution of RS and XPS techniques. The latter represents the oxidation degree for each of the samples and the formation of new bonds promoted by the graphitization of the material. In the RS, two characteristic bands were observed according to the degree of oxidation and the degree of graphitization of the material represented in bands D and G with different relative intensities, suggesting that the samples have different crystallite sizes. This size was described using the Tuinstra-Koenig model, ranging between 18.7 and 25.1 nm. Finally, the bone neoformation observed in the cranial defects of critical size indicates that the F1 and F2 samples, besides being compatible and resorbable, acted as a bridge for bone healing through regeneration. This promoted healing by restoring bone and tissue structure without triggering a strong immune response.

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SUMMARY: CariesCare International ™ practical guide is a tool for the comprehensive assessment and treatment of caries that synthesizes clinical and radiographic diagnosis and risk factors, classifying the severity, progression, and activity of lesions. The objective of this study was to analyze the validity and prediction characteristics of the proposed dental caries classification in the CariesCare International ™ practical guide as a reference through clinical and radiographic evaluation versus histological evaluation. Ninety-seven permanent posterior teeth were evaluated, and clinical and radiographic diagnoses were determined according to parameters defined in the CariesCare International™ guide as a reference. Subsequently, histological evaluation was performed to compare each stage of dental caries progression, and statistical analysis was applied. When comparing the validity and prediction values between radiographic and clinical diagnoses in relation to histological evaluation, a low sensitivity and high specificity relationship was found. The sensitivity and specificity percentages between the clinical and radiographic methods show that the clinical method has a lower number of false negatives. Histological changes in dental tissue were evident from the earliest stages of lesions, even in those not related to the caries process, indicating that the dentist should be careful when deciding on a treatment plan and take into account all variables involved in the caries process, as proposed by the CariesCare International ™ guide.

La guía práctica CariesCare International™ es una herramienta para la evaluación y tratamiento integral de la caries que sintetiza el diagnóstico clínico y radiográfico y los factores de riesgo, clasificando la gravedad, progresión y actividad de las lesiones. El objetivo de este estudio fue analizar la validez y características de predicción de la clasificación de caries dental propuesta en la guía práctica CariesCare International™ como referencia a través de la evaluación clínica y radiográfica versus la evaluación histológica. Se evaluaron noventa y siete dientes posteriores permanentes y se determinaron diagnósticos clínicos y radiográficos según parámetros definidos en la guía CariesCare International™ como referencia. Posteriormente se realizó una evaluación histológica para comparar cada etapa de progresión de la caries dental y se aplicó análisis estadístico. Al comparar los valores de validez y predicción entre los diagnósticos radiológicos y clínicos en relación con la evaluación histológica, se encontró una relación de baja sensibilidad y alta especificidad. Los porcentajes de sensibilidad y especificidad entre el método clínico y radiográfico muestran que el método clínico tiene un menor número de falsos negativos. Los cambios histológicos en el tejido dental fueron evidentes desde las primeras etapas de las lesiones, incluso en aquellas no relacionadas con el proceso de caries, lo que indica que el odontólogo debe tener cuidado al decidir un plan de tratamiento y tener en cuenta todas las variables involucradas en el proceso de caries, tal como propone la guía CariesCare International™.

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Electrospun nanocomposite scaffolds with improved bioactive and biological properties were fabricated from a blend of polycaprolactone (PCL) and starch, and then combined with 5 wt% of calcium oxide (CaO) nanoparticles sourced from eggshells. SEM analyses showed scaffolds with fibrillar morphology and a three-dimensional structure. The hydrophilicity of scaffolds was improved with starch and CaO nanoparticles, which was evidenced by enhanced water absorption (3500 %) for 7 days. In addition, PCL/Starch/CaO scaffolds exhibited major degradation, with a mass loss of approximately 60 % compared to PCL/Starch and PCL/CaO. The PCL/Starch/CaO scaffolds decreased in crystallinity as intermolecular interactions between the nanoparticles retarded the mobility of the polymeric chains, leading to a significant increase in Young's modulus (ca. 60 %) and a decrease in tensile strength and elongation at break, compared to neat PCL. SEM-EDS, FT-IR, and XRD analyses indicated that PCL/Starch/CaO scaffolds presented a higher biomineralization capacity due to the ability to form hydroxyapatite (HA) in their surface after 28 days. The PCL/Starch/CaO scaffolds showed attractive biological performance, allowing cell adhesion and viability of M3T3-E1 preosteoblastic cells. In vivo analysis using a subdermal dorsal model in Wistar rats showed superior biocompatibility and improved resorption process compared to a pure PCL matrix. This biological analysis suggested that the PCL/Starch/CaO electrospun mats are suitable scaffolds for guiding the regeneration of bone tissue.

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Cancer stands as one of the deadliest diseases in human history, marked by an inferior prognosis. While traditional therapeutic methods like surgery, chemotherapy, and radiation have demonstrated success in inhibiting tumor cell growth, their side effects often limit overall benefits and patient acceptance. In this regard, three different graphene oxides (GO) with variations in their degrees of oxidation were studied chemically and tissue-wise. The accuracy of the synthesis of the different GO was verified by robust techniques using X-ray photoelectron spectroscopy (XPS), as well as conventional techniques such as infrared spectroscopy (FTIR), RAMAN spectroscopy, and X-ray diffraction (XRD). The presence of oxygenated groups was of great importance. It affected the physicochemical properties of each of the different graphene oxides demonstrated in the presence of new vibrational modes related to the formation of new bonds promoted by the graphitization of the materials. The toxicity analysis in the Hep-2 cell line of graphene oxide formulations at 250 µg/mL on the viability and proliferation of these tumor cells showed low activity. GO formulations did not show high antibacterial activity against Staphylococcus aureus and Escherichia coli strains. However, the different graphene oxides showed biocompatibility in the subdermal implantation model for 30, 60, and 90 days in the biomodels. This allowed healing by restoring hair and tissue architecture without triggering an aggressive immune response.

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The increasing demand for non-invasive biocompatible materials in biomedical applications, driven by accidents and diseases like cancer, has led to the development of sustainable biomaterials. Here, we report the synthesis of four block formulations using polycaprolactone (PCL), polylactic acid (PLA), and zinc oxide nanoparticles (ZnO-NPs) for subdermal tissue regeneration. Characterization by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) confirmed the composition of the composites. Additionally, the interaction of ZnO-NPs mainly occurred with the C=O groups of PCL occurring at 1724 cm-1, which disappears for F4, as evidenced in the FT-IR analysis. Likewise, this interaction evidenced the decrease in the crystallinity of the composites as they act as crosslinking points between the polymer backbones, inducing gaps between them and weakening the strength of the intermolecular bonds. Thermogravimetric (TGA) and differential scanning calorimetry (DSC) analyses confirmed that the ZnO-NPs bind to the carbonyl groups of the polymer, acting as weak points in the polymer backbone from where the different fragmentations occur. Scanning electron microscopy (SEM) showed that the increase in ZnO-NPs facilitated a more compact surface due to the excellent dispersion and homogeneous accumulation between the polymeric chains, facilitating this morphology. The in vivo studies using the nanocomposites demonstrated the degradation/resorption of the blocks in a ZnO-NP-dependant mode. After degradation, collagen fibers (Type I), blood vessels, and inflammatory cells continue the resorption of the implanted material. The results reported here demonstrate the relevance and potential impact of the ZnO-NP-based scaffolds in soft tissue regeneration.

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This research focused on developing new materials for endodontic treatments to restore tissues affected by infectious or inflammatory processes. Three materials were studied, namely tricalcium phosphate ß-hydroxyapatite (ß-TCP), commercial and natural hydroxyapatite (HA), and chitosan (CS), in different proportions. The chemical characterization using infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis confirmed the composition of the composite. Scanning electron microscopy (SEM) demonstrated that the design and origin of the HA, whether natural or commercial, did not affect the morphology of the composites. In vitro studies using Artemia salina (A. salina) indicated that all three experimental materials were biocompatible after 24 h, with no significant differences in mortality rate observed among the groups. The subdermal implantation of the materials in block form exhibited biocompatibility and biodegradability after 30 and 60 days, with the larger particles undergoing fragmentation and connective tissue formation consisting of collagen type III fibers, blood vessels, and inflammatory cells. The implanted material continued to undergo resorption during this process. The results obtained in this research contribute to developing endodontic technologies for tissue recovery and regeneration.

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Tissue accidents provide numerous pathways for pathogens to invade and flourish, causing additional harm to the host tissue while impeding its natural healing and regeneration. Essential oils (EOs) exhibit rapid and effective antimicrobial properties without promoting bacterial resistance. Clove oils (CEO) demonstrate robust antimicrobial activity against different pathogens. Chitosan (CS) is a natural, partially deacetylated polyamine widely recognized for its vast antimicrobial capacity. In this study, we present the synthesis of four membrane formulations utilizing CS, polyvinyl alcohol (PVA), and glycerol (Gly) incorporated with CEO and nanobioglass (n-BGs) for applications in subdermal tissue regeneration. Our analysis of the membranes' thermal stability and chemical composition provided strong evidence for successfully blending polymers with the entrapment of the essential oil. The incorporation of the CEO in the composite was evidenced by the increase in the intensity of the band of C-O-C in the FTIR; furthermore, the increase in diffraction peaks, as well as the broadening, provide evidence that the introduction of CEO perturbed the crystal structure. The morphological examination conducted using scanning electron microscopy (SEM) revealed that the incorporation of CEO resulted in smooth surfaces, in contrast to the porous morphologies observed with the n-BGs. A histological examination of the implanted membranes demonstrated their biocompatibility and biodegradability, particularly after a 60-day implantation period. The degradation process of more extensive membranes involved connective tissue composed of type III collagen fibers, blood vessels, and inflammatory cells, which supported the reabsorption of the composite membranes, evidencing the material's biocompatibility.

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Autologous bone is the gold standard in regeneration processes. However, there is an endless search for alternative materials in bone regeneration. Xenografts can act as bone substitutes given the difficulty of obtaining bone tissue from patients and before the limitations in the availability of homologous tissue donors. Bone neoformation was studied in critical-size defects created in the parietal bone of 40 adult male Wistar rats, implanted with xenografts composed of particulate bovine hydroxyapatite (HA) and with blocks of bovine hydroxyapatite (HA) and Collagen, which introduces crystallinity to the materials. The Fourier-transform infrared spectroscopy (FTIR) analysis demonstrated the carbonate and phosphate groups of the hydroxyapatite and the amide groups of the collagen structure, while the thermal transitions for HA and HA/collagen composites established mainly dehydration endothermal processes, which increased (from 79 °C to 83 °C) for F2 due to the collagen presence. The xenograft's X-ray powder diffraction (XRD) analysis also revealed the bovine HA crystalline structure, with a prominent peak centered at 32°. We observed macroporosity and mesoporosity in the xenografts from the morphology studies with heterogeneous distribution. The two xenografts induced neoformation in defects of critical size. Histological, histochemical, and scanning electron microscopy (SEM) analyses were performed 30, 60, and 90 days after implantation. The empty defects showed signs of neoformation lower than 30% in the three periods, while the defects implanted with the material showed partial regeneration. InterOss Collagen material temporarily induced osteon formation during the healing process. The results presented here are promising for bone regeneration, demonstrating a beneficial impact in the biomedical field.

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The use of biopolymers for tissue engineering has recently gained attention due to the need for safer and highly compatible materials. Starch is one of the most used biopolymers for membrane preparation. However, incorporating other polymers into starch membranes introduces improvements, such as better thermal and mechanical resistance and increased water affinity, as we reported in our previous work. There are few reports in the literature on the biocompatibility of starch/chicken gelatin composites. We assessed the in vivo biocompatibility of the five composites (T1-T5) cassava starch/gelatin membranes with subdermal implantations in biomodels at 30, 60, and 90 days. The FT-IR spectroscopy analysis demonstrated the main functional groups for starch and chicken gelatin. At the same time, the thermal study exhibited an increase in thermal resistance for T3 and T4, with a remaining mass (~15 wt.%) at 800 °C. The microstructure analysis for the T2-T4 demonstrated evident roughness changes with porosity presence due to starch and gelatin mixture. The decrease in the starch content in the composites also decreased the gelatinization heats for T3 and T4 (195.67, 196.40 J/g, respectively). Finally, the implantation results demonstrated that the formulations exhibited differences in the degradation and resorption capacities according to the starch content, which is easily degraded by amylases. However, the histological results showed that the samples demonstrated almost complete reabsorption without a severe immune response, indicating a high in vivo biocompatibility. These results show that the cassava starch/chicken gelatin composites are promising membrane materials for tissue engineering applications.

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Bone substitutes based on xenografts have been used for a long time in bone regeneration thanks to their inductive capacity for bone tissue regeneration. Some bone-based scaffolds have been modified by adding collagen and other proteins to improve their regenerative capacity and prevent migration and aggregation, especially particles. However, rejection of this graft has been reported due to protein residues caused by poor material preparation. We compared the in vitro and in vivo biological response of two commercial xenografts (InterOss®, F1 and InterOss® Collagen, F2) and a commercial porcine collagen membrane (InterCollagen® Guide, F3) as a rapid degradation control. Fourier Transform Infrared Spectroscopy (FT-IR) analysis evidenced the presence of hydroxyl, orthophosphate, and carbonate groups of the xenografts and amide groups of collagen. Thermogravimetric analysis (TGA) of the xenografts demonstrated their thermal stability and the presence of a few amounts of organic material. The study by differential scanning calorimetry showed the presence of endothermic peaks typical of the dehydration of the xenografts (F1 and F2) and for the collagen membrane (F3), the beginning of structural three-dimensional protein changes. Subsequently, in vitro biocompatibility tests were carried out for the materials with Artemia salina and MTT cell viability with HeLa cells, demonstrating the high biocompatibility of the materials. Finally, in vivo biocompatibility was studied by implanting xenografts in biomodels (Wistar rats) at different periods (30, 60, and 90 days). The F1 xenograft (InterOss) remained remarkably stable throughout the experiment (90 days). F2 (InterOss Collagen) presented a separation of its apatite and collagen components at 60 days and advanced resorption at 90 days of implantation. Finally, the collagen membrane (F3) presented faster resorption since, at 90 days, only some tiny fragments of the material were evident. All the in vivo and in vitro test results demonstrated the biocompatibility of the xenografts, demonstrating the potential of these materials for tissue engineering.