Radiographic Assessment of the Hind Limb Bone Development in Maned Wolves (Chrysocyon brachyurus).

This study aimed to analyse the growth plate fusion and secondary ossification centres of the hind limbs in maned wolves (Chrysocyon brachyurus) using radiographs. Data from three maned wolves estimated to be 3-4 months old were utilised. The right and left hind limbs were radiographed in the mediolateral and craniocaudal views once a month until 11-12 months of age and then every 2-3 months until 18-19 months of age. The growth plates identified in the hind limbs and their closure times were as follows: proximal femur (13-15 months), distal femur (13-15 months), proximal tibia (17-19 months), tibial tuberosity (17-19 months), distal tibia (11-13 months), proximal fibula (13-15 months) and calcaneal tuber (8-9 months). Measurements of the areas of the secondary ossification centres of the proximal epiphysis of the femur, distal epiphysis of the tibia, patella and fibular tarsal bone epiphysis showed a significant difference between the first assessment and 6-7 months. The distal femoral epiphysis, proximal tibial epiphysis and tibial tuberosity showed a statistically significant difference between the first assessment and 7-8 months. The difference in the proximal epiphysis of the fibula was observed between 4 to 5 months and 7 to 8 months. In conclusion, the information obtained regarding the growth plates and secondary ossification centres of the maned wolves may support other studies and help understand the normal patterns of the species.

Evaluation of functioning and associated factors in children and adolescents with osteogenesis imperfecta / Avaliação da funcionalidade e fatores associados de crianças e adolescentes com osteogênese imperfeita

ABSTRACT Objective: The aim of this study was to evaluate the functioning and associated factors in children and adolescents with osteogenesis imperfecta (OI). Methods: This is a cross-sectional study conducted on 30 children and adolescents with OI. Medical records, use of bisphosphonates, socioeconomic status, handgrip strength, balance, joint hypermobility, ambulatory level, and the Pediatric Evaluation of Disability Inventory—Computer Adaptative Test (PEDI-CAT) scores were assessed. Data is presented as mean and standard deviation and Student's t-test or Mann-Whitney U test. Categorical data is presented as frequency and analyzed using Fisher's exact test. Within-group analyses were conducted using ANCOVA or Wilcoxon signed-rank test. Correlations used Kendall's Tau-b test. Results: The participants involved in this study were 6-18 years old. The sample was separated into two groups according to disease severity. The moderate/severe OI group (n=10) presented a lower height and muscular strength than the mild group (n=20). Muscle weakness was observed in all participants with OI when compared with the normal population. No differences were observed between the groups in the PEDI-CAT scores except for the mobility domain. There were correlations between the PEDI-CAT mobility domain and the number of fractures, OI type, weight, and balance; there was also a correlation between the PEDI-CAT daily activities, mobility, responsibility, and social/cognitive domains. Conclusions: The findings suggest that children with moderate/severe forms of OI can achieve the same function levels as children with mild OI. Fractures can have a major influence on the functional level, and treatment should focus on the prevention and rehabilitation of these events when they occur.

RESUMO Objetivo: Avaliar a funcionalidade e fatores associados em crianças e adolescentes com osteogênese imperfeita (OI). Métodos: Estudo transversal com 30 crianças e adolescentes com OI. Foram avaliados prontuários médicos, uso de bisfosfonatos, características socioeconômicas, dinamometria de preensão palmar, equilíbrio, hipermobilidade articular, nível de deambulação e escores do Pediatric Evaluation of Disability Inventory - Computer Adaptative Test (PEDI-CAT). Os dados foram apresentados em média e desvio padrão e comparados por teste t por Mann-Whitney, enquanto os categóricos foram apresentados em frequência e comparados pelo teste exato de Fisher. Análises intragrupos foram realizadas por análise de covariância (ANCOVA) ou Teste de Wilcoxon para postos sinalizados. O teste Tau-b de Kendall foi usado para correlações. Resultados: A idade variou de 6 a 18 anos. A amostra foi dividida em dois grupos de acordo com a gravidade da doença. Casos moderados/graves (n=10) apresentaram menor estatura e força muscular comparadas às dos leves (n=20). Fraqueza muscular foi observada em todos os casos de OI quando comparados à população normal. Não houve diferença nos domínios do PEDI-CAT com exceção do domínio mobilidade. Houve correlação entre o número de fraturas, tipo de OI, peso e equilíbrio e o domínio mobilidade; e entre os domínios Atividades Diárias e Mobilidade e Responsabilidade e Social/cognitivo do PEDI-CAT. Conclusões: Nossos achados sugerem que crianças com OI moderada/severa podem atingir o mesmo nível de funcionalidade que crianças com a forma leve. Fraturas podem ter grande influência no nível de funcionalidade e o tratamento deve enfocar a prevenção e a reabilitação desses eventos.

Socket preservation following tooth extraction: an experimental study comparing ß-Tricalcium phosphate and F18 bioglass in rats.

PURPOSE: To evaluate the efficacy of two third-generation resorbable biomaterials-F18 bioglass and ß-tricalcium phosphate (ß-TCP)-in promoting new bone formation in post-extraction sockets in rats. ß-TCP, a synthetic porous ceramic, is well-established in clinical use, while F18 bioglass, a novel silica based bioglass. METHODS: After extraction of the right upper incisor of 45 rats, the sockets were filled either with F18 or ß-TCP, or left to naturally fill with a blood clot in control group. To quantify new bone formation, five animals from each group were euthanized at 7, 14, and 28 days post-extraction. Subsequently, the hemi-maxillae were analyzed using microtomography and histomorphometry. RESULTS: Two-way ANOVA revealed significant differences in bone neoformation. ß-TCP induced notably higher levels of new bone growth compared to the control group, as evidenced by microtomographic parameters such as bone volume (p = 0.033), bone surface (p = 0.039), and trabecular thickness (p = 0.002). F18 bioglass also induced higher new bone growth compared to the control group, evidenced by bone volume (p = 0.039). Although F18 bioglass did not significantly differ from the control group in other microtomographic parameters of new bone formation, its overall performance was comparable to that of ß-TCP. CONCLUSION: ß-TCP has proven to be effective in promoting new bone formation. Although F18 bioglass did not significantly differ from the control group in several microtomographic parameters, its overall capacity to promote new bone growth was comparable to that of ß-TCP. This outcome supports the use of F18 bioglass as a promising alternative biomaterial for alveolar ridge preservation.

Biological interaction of bioactive polymeric membranes in induced bone defects in rabbit tibias.

The study aimed to evaluate bone repair using three osteoinductive polymers in bone defects created in rabbit tibias. Forty-eight adult rabbits were assessed at various time points: three, seven, fourteen, and thirty days. The groups included a control group (without biomaterial), M1 (Poly L Lactide co Polycaprolactone/Polyethylene Glycol), M2 (Poly L Lactide co Polycaprolactone/Polyethylene Glycol/ß-Tricalcium Phosphate), and M3 (Poly L Lactide co Polycaprolactone/Polyethylene Glycol/nano hydroxyapatite). Histomorphometric analysis was conducted to evaluate new bone formation within and around the bone defect. At 14 (p<0.05) and 30 days (p<0.05), the callus area in the membrane groups, particularly in M3, was also significantly larger than in the control group, indicating the osteoinductive potential of these biomaterials. The callus consisted of both bone and cartilaginous matrix, suggesting a robust activation of endochondral ossification. The number of osteoclast was higher in the membrane groups, especially at 14 days in the M3 group, indicating increased bone remodeling activity. The membranes were not fully absorbed by 30 days, creating a space between the defect and the periosteum. In conclusion, all three membranes showed significant chondro and osteoinductive potential, with the membrane containing nano-hydroxyapatite demonstrating the most pronounced potential.

Anti-Osteoclastogenesis Potential of Cocoa Pod Husk (Theobroma cacao L.) Extract: In Silico and In Vivo Study.

Periodontitis is a common chronic inflammatory disease characterized by alveolar bone loss. The high polyphenol content in cocoa pod husk (Theobroma cacao L) has the potential to influence bone metabolism and contribute to the inhibition of bone resorption. The aim of this study was to analyze the anti-osteoclastogenesis potential of cocoa pod husk (Theobroma cacao L.) in both in silico and in vivo study. An analysis of the anti-osteoclastogenesis potential of T. cacao bioactive compounds was conducted using molecular docking simulations. Thirty male Wistar rats (Rattus novergicus) were randomly assigned to control negative groups (placebo gel), control positive groups (2% doxycycline gel), and treatment groups (10% cocoa pod husk (CPH) extract gel), with measurements taken on days 7 and 14. Wistar rats were induced with 0.05 ml of P. gingivalis at a concentration of 2x109 CFU/ml intrasulcularly in the maxillary molar to achieved in periodontitis. The number of osteoclasts was observed by hematoxylin and eosin staining, the level of TNF-α was assessed by enzyme-linked immunosorbent assay, and the expression of RANKL was evaluated by immunohistochemical staining. Data were analyzed using One-way ANOVA to examine the differences between the groups. The in silico study showed that the catechin, epicatechin, quercetin, and procyanidin B2 had a strong binding affinity for TNF-α and RANKL. Administration of 10% CPH reduced the number of osteoclasts (p<0.05), TNF-α level on days 7 and 14 (p<0.05), and RANKL expression on day 7 (p<0.05) in experimental rats with periodontitis. Administering 10% CPH inhibited osteoclastogenesis in the experimental periodontitis rats.

A preliminary study of cell-based bone tissue engineering into 3D-printed ß-tricalcium phosphate scaffolds and polydioxanone membranes.

Treatment of complex craniofacial deformities is still a challenge for medicine and dentistry because few approach therapies are available on the market that allow rehabilitation using 3D-printed medical devices. Thus, this study aims to create a scaffold with a morphology that simulates bone tissue, able to create a favorable environment for the development and differentiation of osteogenic cells. Moreover, its association with Plenum Guide, through cell-based tissue engineering (ASCs) for guided bone regeneration in critical rat calvarial defects. The manufacturing and characterization of 3D-printed ß-TCP scaffolds for experimental surgery was performed. Nine male rats were divided into three groups: ß-TCP + PDO membrane (TCP/PG), ß-TCP/ASCs + PDO membrane (TCPasc/PG), and ß-TCP/ASCs + PDO membrane/ASCs (TCPasc/PGasc). A surgical defect with a 5-mm diameter was performed in the right parietal bone, and the defect was filled with the 3D-printed ß-TCP scaffold and PDO membrane with or without ASCs. The animals were euthanized 7, 14, and 30 days after the surgical procedure for histomorphometric and immunolabeling analyses. 3D-printed ß-TCP scaffolds were created with a 404 ± 0.0238 µm gyroid macro-pore and, the association to cell-based therapy promotes, especially in the TCPasc/PGasc group, a bone area formation at the defect border region and the center of the defect. The use of 3D-printed ß-TCP scaffolds and PDO membranes associated with cell-based therapy could improve and accelerate guided bone regeneration, promoting an increase in osteogenic capacity and reducing the time involved in the bone formation process. Moreover, using ASCs optimized the bioceramics by increasing its osteoinductive and osteoprogenitor capacity and, even with the resorption of the printed scaffold, aided as a scaffold for mesenchymal cell differentiation, as well as in bone tissue formation.

The Influence of Physiological Blood Clot on Osteoblastic Cell Response to a Chitosan-Based 3D Scaffold-A Pilot Investigation.

BACKGROUND: The use of ex vivo assays associated with biomaterials may allow the short-term visualization of a specific cell type response inserted in a local microenvironment. Blood is the first component to come into contact with biomaterials, providing blood clot formation, being substantial in new tissue formation. Thus, this research investigated the physiological blood clot (PhC) patterns formed in 3D scaffolds (SCAs), based on chitosan and 20% beta-tricalcium phosphate and its effect on osteogenesis. Initially, SCA were inserted for 16 h in rats calvaria defects, and, after that, osteoblasts cells (OSB; UMR-106 lineage) were seeded on the substrate formed. The groups tested were SCA + OSB and SCA + PhC + OSB. Cell viability was checked by MTT and mineralized matrix formation in OSB using alizarin red (ARS). The alkaline phosphatase (ALP) and bone sialoprotein (BSP) expression in OSB was investigated by indirect immunofluorescence (IF). The OSB and PhC morphology was verified by scanning electron microscopy (SEM). RESULTS: The SCA + PhC + OSB group showed greater cell viability (p = 0.0169). After 10 days, there was more mineralized matrix deposition (p = 0.0365) and high ALP immunostaining (p = 0.0021) in the SCA + OSB group. In contrast, BSP was more expressed in OSB seeded on SCA with PhC (p = 0.0033). CONCLUSIONS: These findings show the feasibility of using PhC in ex vivo assays. Additionally, its inclusion in the experiments resulted in a change in OSB behavior when compared to in vitro assays. This "closer to nature" environment can completely change the scenario of a study.

Variante de novo en el gen COL1A1 asociado a enfermedad genética huérfana: Osteogénesis Imperfecta tipo I

Introducción: La Osteogénesis Imperfecta (OI), se conoce como un trastorno caracterizado por debilidad ósea y alto riesgo de fracturas, de origen genético, conocido como enfermedad de los huesos de cristal. Tiene alta carga de morbilidad- mortalidad, asociada a presencia de dolor y limitación funcional para quienes la presentan. La Fragilidad que se presenta en los huesos se ha descrito por variantes estructurales de los genes del Colágeno 1 (COL1A1 y COL1A2) entre otros, tiene prevalencia estimada entre 1/15,000 y 1/20,000 casos en recién nacidos vivos. El diagnóstico oportuno de la enfermedad permite iniciar alternativas terapéuticas dirigidas, prevenir complicaciones, dar asesoría genética sobre riesgo de heredabilidad, pautas de seguimiento, pronóstico acercándonos a la medicina personalizada. Presentación de Caso: Paciente femenina de 8 años de edad, con historia clínica de varias fracturas secundarias a traumas leves (metatarso pie izquierdo, dedo de mano derecha y pie ipsilateral), con otros diagnósticos de otoesclerosis bilateral con requerimiento de audífonos, pubertad precoz, al examen físico con dismorfias faciales dadas por frente amplia prominente, escleras leve tinte azul, dientes irregulares con desgaste de la dentina, hiperlaxitud ligamentaria, no compromiso cognitivo­conductual y sin otras alteraciones. Dentro del abordaje paraclínico con hipercalcemia, hipercalciuria e hiperfosfatemia, resto de paraclínicos de evaluación de metabolismo óseo en rango de referencia normal, dentro de los estudios imagenológicos con radiografía de columna vertebral total con leve escoliosis dextro convexa lumbar, y anterolistesis de L5 sobre S1, y las radiografías que evidenciaban zonas de consolidación por fracturas descritas previamente, ecografía renal y vías urinarias con leve ectasia bilateral y residuo del 15% en el vaciamiento vesical. Paciente producto de padres no consanguíneos, sin historial familiar conocido de enfermedades genéticas del tejido conectivo. Por los antecedentes clínicos y examen físico ante sospecha de Colagenopatía se solicitó estudio molecular secuenciación + CNVs (del inglés Copy Number Variations) mediante NGS (del inglés Next-Generation Sequencing) para genes relacionados con colagenopatías. Resultados: Estudio molecular secuenciación + CNVs mediante NGS para genes relacionados con colagenopatías en el cual se detecta una variante patogénica heterocigota c.3652G>A; p.Ala1218Thr en el gen COL1A1, lo cual da soporte al diagnóstico de osteogénesis imperfecta tipo I de herencia autosómica dominante. Conclusión: La osteogénesis imperfecta es una enfermedad de origen genético, rara, reportada desde hace muchos años, con variabilidad fenotípica, endotípica y genotípica, por lo cual emprender estrategias de identificación temprana impacta en la historia natural de la enfermedad contribuyendo al desarrollo de una medicina preventiva, personalizada, de predicción, precisión, participativa en aras de poderse realizar a nivel poblacional; por medio de este caso se pretende concientizar sobre la importancia de un diagnóstico precoz y correlación genotipo-fenotipo que permita instaurar acciones dirigidas, conocer mecanismos fisiopatológicos específicos, sub - clasificar las colagenopatías, contribuyendo en el aumento del conocimiento médico- científico sobre la expresión de la enfermedad y la tendencia a la hiperpersonalización. (provisto por Infomedic International)

Introduction: Osteogenesis Imperfecta (OI), is known as a disorder characterized by bone weakness and high risk of fractures, of genetic origin, known as brittle bone disease. It has a high burden of morbidity and mortality, associated with the presence of pain and functional limitation for those who present it. Fragility in the bones has been described by structural variants of the Collagen 1 genes (COL1A1 and COL1A2) among others, with an estimated prevalence between 1/15,000 and 1/20,000 cases in live newborns. The timely diagnosis of the disease allows to initiate targeted therapeutic alternatives, prevent complications, provide genetic counseling on risk of heritability, follow-up guidelines, prognosis approaching personalized medicine. Case Presentation: Female patient 8 years old, with clinical history of several fractures secondary to mild trauma (metatarsal left foot, right hand finger and ipsilateral foot), with other diagnoses of bilateral otosclerosis requiring hearing aids, precocious puberty, on physical examination with facial dysmorphias given by prominent broad forehead, mild blue tint sclerae, irregular teeth with dentin wear, ligament hyperlaxity, no cognitive-behavioral compromise and no other alterations.Within the paraclinical approach with hypercalcemia, hypercalciuria and hyperphosphatemia, other paraclinical evaluation of bone metabolism in normal reference range, within the imaging studies with total spine radiography with mild lumbar dextro convex dextro scoliosis, and anterolisthesis of L5 over S1, and radiographs showing areas of consolidation by fractures previously described, renal and urinary tract ultrasound with mild bilateral ectasia and 15% residue in bladder emptying. The patient was the product of non-consanguineous parents, with no known family history of genetic connective tissue diseases. Due to the clinical history and physical examination, a molecular sequencing + CNVs (Copy Number Variations) study was requested by NGS (Next-Generation Sequencing) for genes related to collagenopathies. Results: Molecular study sequencing + CNVs by NGS for genes related to collagenopathies in which a heterozygous pathogenic variant c.3652G>A; p.Ala1218Thr in the COL1A1 gene is detected, which supports the diagnosis of osteogenesis imperfecta type I of autosomal dominant inheritance. Conclusion: Osteogenesis imperfecta is a disease of genetic origin, rare, reported for many years, with phenotypic, endotypic and genotypic variability, so that undertake early identification strategies impact on the natural history of the disease contributing to the development of preventive medicine, personalized, predictive, accurate, participatory in order to be performed at the population level; through this case it is intended to raise awareness of the importance of early diagnosis and genotype-phenotype correlation that allows to establish targeted actions, to know specific pathophysiological mechanisms, to sub-classify collagenopathies, contributing to the increase of medical-scientific knowledge about the expression of the disease and the tendency to hyper-personalization. (provided by Infomedic International)

Una revolución genómica: Avances en salud y conocimiento en América Latina

Editorial: Investigación pionera en genética y genómica para el progreso regional Al entrar en el segundo año de existencia de la Revista de Genética y Genómica Clínica, se puede evidenciar muchas expectativas académicas en materia de investigación. Es extraordinario ver lo lejos que hemos llegado en tan poco tiempo. Esta revista nació de la visión de reunir lo mejor de la investigación en genética y genómica y tener un impacto significativo en la asistencia sanitaria y la educación en nuestra región. Esa visión se está haciendo realidad, gracias a las increíbles contribuciones de nuestros autores y al apoyo de nuestros lectores. Este número presenta una impresionante selección de artículos que ponen de relieve el poder de la investigación genómica para transformar vidas. Nuestra sección de Casos clínicos incluye historias de enfermedades raras y complejas en las que la genética ha desempeñado un papel fundamental: En el Síndrome Hipotónico como manifestación de una enfermedad ultra-rara causada por una nueva variante de novo en el gen PLA2G6, vemos el intrincado proceso de resolución de un misterio diagnóstico y sus profundas implicaciones para el cuidado del paciente. Variante de novo en el gen COL1A1 asociada a una enfermedad genética huérfana: La osteogénesis imperfecta de tipo I arroja luz sobre una enfermedad rara que supone un reto tanto para las familias como para los médicos, pero que también presenta oportunidades para tratamientos específicos. Variante genética de novo en la Encefalopatía Epiléptica: la importancia de un diagnóstico específico pone de relieve cómo la identificación de una causa genética puede conducir a estrategias terapéuticas más eficaces. Por último, Detección de una variante genética en el Síndrome de Apert examina una afección que, aunque poco frecuente, tiene importantes implicaciones terapéuticas y para el desarrollo. Nuestros artículos de revisión, Avances y Perspectivas de las patologías genéticas en el siglo XXI, ofrecen valiosas panorámicas de las tendencias actuales y las posibilidades emergentes, sirviendo de guía esencial para cualquiera que se dedique a la investigación genética o a la práctica clínica. Por qué es importante? La genética y la genómica ya no es una disciplina rara, sino que están transformando nuestra forma de entender la salud y la enfermedad. En nuestra región, este conocimiento tiene el potencial de hacer frente a las desigualdades en la atención sanitaria, garantizando que los avances en la medicina de precisión beneficien a todas las comunidades, no sólo a unos pocos elegidos. Igualmente apasionante es el papel que la genética puede desempeñar en la educación. Es esencial formar a la próxima generación de profesionales sanitarios e investigadores para que aprovechen el poder de la genómica. Al invertir en educación, estamos sentando las bases para un futuro en el que la medicina genómica esté plenamente integrada en la práctica diaria. Esta revista es un testimonio de lo que es posible cuando trabajamos juntos: científicos, médicos, educadores y responsables políticos. Es una invitación a seguir superando los límites, a plantear preguntas difíciles y a encontrar respuestas que importen. Gracias por acompañarnos en este viaje. Espero que este número les inspire, les haga reflexionar y les recuerde el increíble potencial de la genética y la genómica para crear un mundo más sano y equitativo.   (provisto por Infomedic International)

Editorial: Pioneering Genetics and Genomics Research for Regional Progress As we enter the second year of the Genetics and Clinical Genomics Journal, there is much anticipation for the future of this field in the region. It's remarkable to see how far we've come in such a short time. This journal was born from a vision to bring together the best of genetics and genomics research and make a meaningful impact on healthcare and education in our region. That vision is becoming a reality, thanks to the incredible contributions of our authors and the support of our readers. This issue features an impressive selection of articles that highlight the power of genomic research to transform lives. Our Case Reports section includes stories of rare and complex conditions where genetics has played a critical role: In Hypotonic syndrome as a manifestation of an ultra-rare disease caused by a novel de novo variant in the PLA2G6 gene, we see the intricate process of solving a diagnostic mystery and its profound implications for patient care. De novo variant in the COL1A1 gene associated with an orphan genetic disease: Type I osteogenesis imperfecta sheds light on a rare condition that challenges both families and clinicians but also presents opportunities for targeted treatments. De novo genetic variant in Epileptic Encephalopathy: The importance of specific diagnosis highlights how pinpointing a genetic cause can lead to more effective therapeutic strategies. Finally, Detection of a genetic variant in Apert syndrome examines a condition that, while rare, has significant developmental and therapeutic implications. Our Review Articles, Literature review and Advances and perspectives on genetic pathologies in the 21st century, provide valuable overviews of current trends and emerging possibilities, serving as essential guides for anyone engaged in genetic research or clinical practice. Why This Matters Genetics and genomics are no longer rare disciplines they're reshaping how we understand health and disease. In our region, this knowledge has the potential to address long-standing healthcare inequities, ensuring that advances in precision medicine benefit all communities, not just a select few. Equally exciting is the role genetics can play in education. Training the next generation of healthcare providers and researchers to harness the power of genomics is essential. By investing in education, we're laying the groundwork for a future where genomic medicine is fully integrated into everyday practice. This journal is a testament to what's possible when we work together­scientists, clinicians, educators, and policymakers. It's an invitation to keep pushing boundaries, asking tough questions, and finding answers that matter. Thank you for joining us on this journey. I hope you find this issue inspiring, thought-provoking, and a reminder of the incredible potential within genetics and genomics to create a healthier, more equitable world. (provided by Infomedic International)

Lithium-containing 45S5 Bioglass-derived glass-ceramics have antioxidant activity and induce new bone formation in a rat preclinical model of type 1 diabetes mellitus.

Diabetes mellitus (DM) has been associated with complications that affect the skeletal system, such as alterations in bone repair, osteoporosis, and an increased risk of fractures. In this context, the use of biomaterials able to promote osteogenic differentiation and, at the same time, limit the oxidative stress induced by DM offers a novel perspective to ensure the repair of diabetic bone tissue. Since lithium (Li) has been recently identified as a biologically active ion with osteogenic and antioxidant properties, the localized and controlled release of Li ions from bioactive glass-ceramic materials represents a promising therapeutic alternative for the treatment of bone lesions in DM. Thus, the aim of this study was to evaluate the potential osteogenic and antioxidant effects of glass-ceramic microparticles derived from a 45S5-type bioactive glass (Bioglass) containing (% by weight) 45% SiO2, 24.5% Na2O, 24.5% CaO, and 6% P2O5, in which Na2O was partially substituted by 5% of Li2O (45S5.5Li), in an experimental model of type 1 DM (DM1). The results obtained demonstrate, for the first time, that both 45S5 and 45S5.5Li glass-ceramic microparticles possess antioxidant activity and stimulate bone formationin vivoboth under physiological conditions and under experimental DM1 in rats. In this sense, they would have potential application as inorganic osteogenic agents in different strategies of bone tissue regenerative medicine.

7-Ketocholesterol Effects on Osteogenic Differentiation of Adipose Tissue-Derived Mesenchymal Stem Cells.

Some oxysterols were shown to promote osteogenic differentiation of mesenchymal stem cells (MSCs). Little is known about the effects of 7-ketocholesterol (7-KC) in this process. We describe its impact on human adipose tissue-derived MSC (ATMSC) osteogenic differentiation. ATMSCs were incubated with 7-KC in osteogenic or adipogenic media. Osteogenic and adipogenic differentiation was evaluated by Alizarin red and Oil Red O staining, respectively. Osteogenic (ALPL, RUNX2, BGLAP) and adipogenic markers (PPARƔ, C/EBPα) were determined by RT-PCR. Differentiation signaling pathways (SHh, Smo, Gli-3, ß-catenin) were determined by indirect immunofluorescence. ATMSCs treated with 7-KC in osteogenic media stained positively for Alizarin Red. 7-KC in adipogenic media decreased the number of adipocytes. 7-KC increased ALPL and RUNX2 but not BGLAP expressions. 7-KC decreased expression of PPARƔ and C/EBPα, did not change SHh, Smo, and Gli-3 expression, and increased the expression of ß-catenin. In conclusion, 7-KC favors osteogenic differentiation of ATMSCs through the expression of early osteogenic genes (matrix maturation phase) by activating the Wnt/ß-catenin signaling pathway, while inhibiting adipogenic differentiation. This knowledge can be potentially useful in regenerative medicine, in treatments for bone diseases.

Effect of laser photobiomodulation combined with hydroxyapatite nanoparticles on the osteogenic differentiation of mesenchymal stem cells using artificial intelligence: An in vitro study.

AIM: To evaluate in vitro the effect of laser photobiomodulation (PBM) combined or not with 30-nm hydroxyapatite nanoparticles (HANp), on the osteogenic differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) by morphometric analysis using artificial intelligence programs (TensorFlow and ArcGIS). METHODS: UC-MSCs were isolated and cultured until 80% confluence was reached. The cells were then plated according to the following experimental groups: G1 -control (DMEM), G2 -BMP-2, G3 -BMP-7, G4 -PBM (660 nm, 10 mW, 2.5 J/cm2, spot size of 0.08 cm2), G5 -HANp, G6 -HANp + PBM, G7 -BMP-2 + PBM, and G8 -BMP-7 + PBM. The MTT assay was used to analyze cell viability at 24, 48 and 72 h. Osteogenic differentiation was assessed by Alizarin Red staining after 7, 14 and 21 days. For morphometric analysis, areas of osteogenic differentiation (pixel2) were delimited by machine learning using the TensorFlow and ArcGIS 10.8 programs. RESULTS: The results of the MTT assay showed high rates of cell viability and proliferation in all groups when compared to control. Morphometric analysis revealed a greater area of osteogenic differentiation in G5 (HANp = 142709,33±36573,39) and G6 (HANp + PBM = 125452,00±24226,95) at all time points evaluated. CONCLUSION: It is suggested that HANp, whether combined with PBM or not, may be a promising alternative to enhance the cellular viability and osteogenic differentiation of hUC-MSCs.

1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential.

OBJECTIVE: Periodontal dental ligament mesenchymal stem cells (PDLMSCs) play a major role in periodontal tissue regeneration by the neoformation of root cementum and alveolar bone. These cells are highly heterogeneous, and many present low potential to renovate the hard tissue damaged by periodontal disease. A previous study found that the low osteoblast/cementoblast (O/C) differentiation potential of PDLMSCs is related to high asporin (ASPN) expression, which was identified as a negative regulator of PDL cells differentiation and mineralization, suppressing BMP-2-induced O/C differentiation. This study aimed to investigate whether 1,25(OH)2D3 treatment could stimulate the O/C differentiation of periodontal ligament mesenchymal progenitor cells characterized as low osteoblast potential (LOP), by asporin and bone morphogenetic protein-2 alteration. METHODOLOGY: Three LOP cell populations were cultured in standard medium (CONTROL), osteogenic medium (OM), and osteogenic medium associated with 1 nM of 1,25(OH)2D3 (OM + VD). The following assays were performed: 1) MTT to evaluate metabolic activity; 2) gene expression for asporin (ASPN), bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and vitamin D receptor (VDR) using qRT-PCR; 3) BMP-2 extracellular expression; and 4) quantification of mineralized nodule deposition by Alizarin Red Staining. Data were subjected to two-way ANOVA and Tukey's test (P<0.05). RESULTS: The results showed that the 1,25(OH)2D3 treatment did not affect the cell viability, as demonstrated by metabolic activity increase over the 10 days in culture. After 14 days of 1,25(OH)2D3 treatment, the mRNA levels for ASPN and VDR decreased (P<0.05), while BMP-2 transcripts and extracellular expression increased (P<0.05). In parallel, RUNX2, ALP, and OCN gene expression was upregulated by 1,25(OH)2D3 treatment, resulting in an increase of mineral nodule deposition in vitro (P<0.05). CONCLUSIONS: These data show that 1,25(OH)2D3 improves osteoblast/cementoblast differentiation of low osteoblast potential accompanied by alterations in ASPN and BMP-2 expression.

* This article is derived from a master's dissertation.

Fish: a new xenograft source for maxillary sinus lifting.

OBJECTIVE: Although autogenous grafting is accepted as the gold standard in intraoral grafting, xenogenous grafts are frequently used in sinus lift surgeries due to their osteoinductive and osteoconductive properties. This study aimed to investigate the efficacy of fish spine-derived xenogenic grafts in sinus augmentation surgery. MATERIAL AND METHODS: In this study, a fish spine-derived xenogenic graft was produced for comparison with autogenous graft and bovine derived xenogenic grafts. Twenty-one New Zealand rabbits were used. Autogenous grafts (AG- Group 1), as well as bovine-derived (bHAP - Group 2) and fish spine-derived (fHAP - Group 3) xenogenic grafts were placed in the right and left sinuses of rabbits. The animals were sacrificed at the 4th and 8th weeks. New bone formation (NBF) was evaluated through histological examination, while bone volume (BV), new bone surface/bone volume (BS-BV), new bone surface/tissue volume (BS-TV), and trabecular separation (Tb-Sp) were assessed via Micro-CT. Statistical significance was considered at p<0.05. RESULTS: Histological examination revealed a significant difference in NBF between AG-bHAP (p<0.001), as well as between fHAP-bHAP (p<0.001) in the fourth-week group. No significant difference was found in the eighth-week group (p=0.130). In the eighth-week group, a statistically significant difference was found between fHAP and bHAP in terms of BV. (p=0.007). CONCLUSION: Although both graft materials used in this study showed positive effects on bone regeneration, fHAP and AG presented similar effects on bone regeneration and were superior to bHAP.

Investigating the Biological Efficacy of Albumin-Enriched Platelet-Rich Fibrin (Alb-PRF): A Study on Cytokine Dynamics and Osteoblast Behavior.

The development of effective biomaterials for tissue regeneration has led to the exploration of blood derivatives such as leucocyte- and platelet-rich fibrin (L-PRF). A novel variant, Albumin-Enriched Platelet-Rich Fibrin (Alb-PRF), has been introduced to improve structural stability and bioactivity, making it a promising candidate for bone regeneration. This study aimed to evaluate Alb-PRF's capacity for cytokine and growth factor release, along with its effects on the proliferation, differentiation, and mineralization of human osteoblasts in vitro. Alb-PRF membranes were analyzed using histological, scanning electron microscopy, and fluorescence microscopy techniques. Cytokine and growth factor release was quantified over seven days, and osteoinductive potential was evaluated with MG-63 osteoblast-like cells. Structural analysis showed Alb-PRF as a biphasic, highly cellularized material that releases lower levels of inflammatory cytokines and higher concentrations of platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) compared to L-PRF. Alb-PRF exhibited higher early alkaline phosphatase activity and in vitro mineralization (p < 0.05) and significantly increased the OPG/RANKL mRNA ratio (p < 0.05). These results indicate that Alb-PRF has promising potential as a scaffold for bone repair, warranting further in vivo and clinical assessments to confirm its suitability for clinical applications.

Local application of melatonin associated or not to xenogeneic material, in critical defects of rat calvaria.

Melatonin (MLT) is a hormone that can stimulate bone formation and inhibit bone resorption, among other functions. Aim: To evaluate the effect on new bone formation of MLT applied locally to critical defects created in the calvaria of rats, compared to the effect of Bio-Oss® xenogeneic bone substitute (BO), by analyzing histomorphometry, microtomography and gene expression. Materials and Method: Two critical defects (5.0 mm in diameter) were created in the calvaria of 36 adults male Wistar rats. The rats were divided randomly into two groups: a test group, in which one of the defects was filled with MLT, and the other with MLT with Bio-Oss® (MLTBO), and a control group, in which one of the defects was filled only with the clot (C), and the other with BO. The rats were euthanized 30 days after surgery. Samples of the calvaria containing the critical defects were collected for analysis by histomorphometry, microtomography, and the expression of the genes for type I collagen (COL-I), osteopontin (OPN) and bone morphogenetic protein 2 (BMP-2). Results: A qualitative improvement was observed in bone healing when MLT was used, though there was no statistical difference in the quantification of newly formed bone (p>0.05). Micro-CT showed that bone volume was significantly smaller in absence of BO (p=0.006). Bone trabeculae thickness (p=0.590) and number (p=0.150) were not significantly affected by MLT. Regarding the expression of the genes COL-I, OPN and BMP-2, no significant differences were observed between the MLT, BO and MLTBO groups. Conclusion: Topical application of MLT resulted in a qualitative improvement in bone healing, although it did not affect bone formation quantitatively. In the absence of BO, less bone volume and less bone trabecular thickness were observed.

A melatonina (MLT) é um hormônio sintetizado e secretado pela glândula pineal, e que, dentre outras atividades e funções, tem capacidade de estimular a formação e inibir a reabsorção óssea. Objetivo: avaliar o efeito da aplicação local do hormônio MLT na neoformação óssea, em defeitos críticos produzidos na calvária de ratos, por meio de análise histomorfométrica, microtomográfica e de biologia molecular, comparando-a com um substituto ósseo de origem xenogênica (Bio-Oss®). Materiais e Método: foram utilizados 36 ratos Wistar machos adultos, nos quais foram produzidos dois defeitos críticos de 5,0 mm de diâmetro cada, que receberam diferentes tratamentos alocados de forma randomizada: no grupo controle, os animais tiveram um dos defeitos preenchidos apenas com coágulo sanguíneo (C) e o outro com substituto ósseo xenógeno (Bio-Oss®, BO); no grupo teste, um dos defeitos foi preenchido apenas com MLT e, o outro, recebeu a associação da MLT ao material sintético xenógeno (MLTBO). Todos os animais foram eutanasiados após 30 dias do pós-operatório. As amostras das calotas contendo os defeitos críticos foram coletadas para análises histomorfométricas, microtomográficas e da expressão gênica de colágeno do tipo I (COL-I), osteopontina (OPN) e proteína óssea morfogenética 2 (BMP-2), por meio de PCR em tempo real. Resultados: Após análise dos dados pode-se observar que não houve diferença estatística na quantificação de neoformação óssea (p>0.05), porém, melhora qualitativa foi observada na cicatrização, quando a MLT foi utilizada. Quanto aos par®metros microtomográficos, foi observado que com ou sem MLT, o volume ósseo foi significativamente menor na ausência de BO. A espessura (p = 0,590) e número (p = 0,150) de trabéculas não foram significativamente afetados pelo uso da MLT. Quanto à expressão gênica de COL-I, OPMN e BMP-2, não foram observadas diferenças significativas entre os grupos MLT, BO e MLTBO. Conclusão: conclui-se que a aplicação tópica de MLT, associada ou não ao BO não afetou quantitativamente a neoformação óssea, porém resultou em uma melhora qualitativa na cicatrização. Adicionalmente, na ausência de BO foi observada menor volume ósseo e menor espessura das trabéculas.

DNMT3A transcriptionally downregulated by KLF5 alleviates LPS-induced inflammatory response and promotes osteogenic differentiation in hPDLSCs.

BACKGROUND AND OBJECTIVE: Periodontitis is an inflammatory disease typically characterized by the destruction of periodontal tissues and complicated etiology. DNA methyltransferase 3A (DNMT3A) has been implicated in possessing pro-inflammatory properties. This study sought to explore the role of DNMT3A in periodontitis and its relevant mechanism. METHODOLOGY: Lipopolysaccharide (LPS) was used to induce inflammation in human periodontal ligament stem cells (hPDLSCs). DNMT3A and KLF5 expressions were detected using RT-qPCR and western blot. The levels of inflammatory cytokines and inflammation-related proteins were detected using ELISA and western blot. NF-κB p65 expression was detected using immunofluorescence (IF) assay, while osteogenic differentiation was assessed using ALP assay and ARS staining. Western blot was used to measure the protein contents associated with osteogenic differentiation. DNMT3A activity was detected using luciferase report assay and chromatin immunoprecipitation (ChIP) was used to verify the interaction between KLF5 and DNMT3A. RESULTS: DNMT3A expression increased in LPS-induced hPDLSCs. Silencing DNMT3A suppressed the LPS-induced inflammation in hPDLSCs, while promoting osteogenic differentiation. It was also found that transcriptional factor KLF5 could bind to DNMT3A promoters and regulate DNMT3A expression. Rescue experiments showed that KLF5 interference partially counteracted the inhibitory impacts of DNMT3A deficiency on inflammation and the promotive effects on osteogenic differentiation in LPS-induced hPDLSCs. CONCLUSION: DNMT3A, when transcriptionally downregulated by KLF5, could alleviate LPS-challenged inflammatory responses and facilitate osteogenic differentiation in hPDLSCs.

Enhancing osteoblast differentiation and bone repair: The priming effect of photobiomodulation on adipose stromal cells.

Cellular therapy using adipose tissue-derived mesenchymal stromal cells (at-MSCs) has garnered attention for the treatment of bone defects. Therefore, preconditioning strategies to enhance the osteogenic potential of at-MSCs could optimize cell therapy outcomes, and photobiomodulation (PBM) therapy has emerged as an effective, noninvasive, and low-cost alternative. This study explored the impacts of PBM on at-MSCs differentiation and the subsequent repair of bone defects treated with cell injection. Rat at-MSCs were cultured and irradiated (at-MSCsPBM) following the PBM protocol (660 nm; 20 mW; 0.714 W/cm2; 0.14 J; 5 J/cm2). Cellular differentiation was assessed based on the expression of gene and protein markers. Reactive oxygen species (ROS) were detected using fluorescence. At-MSCsPBM were injected into 5-mm calvarial lesions, and bone formation was analyzed using micro-CT and histological evaluations. At-MSCs were used as control. Data were analyzed using the ANOVA or t-test. At-MSCsPBM exhibited high levels of gene and protein runt-related transcription factor-2 (Runx2) and alkaline phosphatase (Alp) expression. PBM increased ALP activity and significantly reduced ROS levels. In addition, PBM increased the expression of Wnt pathway-associated genes. In vivo, there was an increase in the morphometric parameters, including bone volume, percentage of bone volume, bone surface area, and trabecular number, in at-MSCsPBM-treated defects compared with those in the control. These findings suggest that PBM enhances the osteogenic potential of at-MSCs, thereby supporting the advancement of improved cellular therapies for bone regeneration.

Mesenchymal Stem Cells and Tissue Bioengineering Applications in Sheep as Ideal Model.

The most common technologies in tissue engineering include growth factor therapies; metal implants, such as titanium; 3D bioprinting; nanoimprinting for ceramic/polymer scaffolds; and cell therapies, such as mesenchymal stem cells (MSCs). Cell therapy is a promising alternative to organ grafts and transplants in the treatment of numerous musculoskeletal diseases. MSCs have increasingly been used in generative medicine due to their specialized self-renewal, immunomodulation, multiplication, and differentiation properties. To further expand the potential of these cells in tissue repair, significant efforts are currently dedicated to the production of biomaterials with desirable short- and long-term biophysical properties that can aid the differentiation and expansion of MSCs. Biomaterials support MSC differentiation by modulating their characteristics, such as composition, mechanical properties, porosity, and topography. This review aimed to describe recent MSC approaches, including those associated with biomaterials, from experimental, clinical, and preclinical studies with sheep models.

Forensic age estimation from ossification centres: a comparative investigation of imaging and physical methods.

Age estimation is a crucial component of human identification in forensic science. It has a vital role in forensic anthropology, including examinations of skeletal remains, disaster victim identification, and locating missing individuals. Present communication focuses on the age estimation through the examination of ossification centers of bones and its significance in identifying the age of 18 years old, a recognized age of majority in many countries. The process of ossification is integral to biological development and serves as critical standard for age estimation in forensic identification. This study reviews relevant literature from well-known databases such as PubMed, Scopus, Web of Science, and ScienceDirect. Additionally, the present review elaborates various classification methods used by authors to classify the stages of ossification centers of bones. The objective of this communication is to assess the effectiveness of both imaging and physical methods for age estimation and to provide a critical comparison to determine the superior approach. The findings suggest that imaging methods are more reliable for the estimation of age from ossification centers. Staging methods introduced by Schmeling et al, Kellinghaus et al, Dedouit et al, Vieth et al, and Kvist et al. are found to be the best methods for age estimation.