Bone Tissue Samples Research Articles

Zoledronic acid-treated rats show altered jaw histology and gene expression in non-exposed MRONJ

BackgroundDiagnosis and management of medication-related osteonecrosis of the jaws (MRONJ) prior to clinical exposure induced by trauma may lead to improved patient management. Currently, few studies have examined early histologic and molecular MRONJ-related changes in the jaws. PurposeThis study aimed to identify histological and gene expression changes in the maxilla and mandible of Sprague-Dawley (SD) rats after zoledronic acid (ZA) treatment. Study Design, Setting, and SampleThis was an in-vivo animal study. The experiments were conducted in the laboratory at the Stomatology Hospital of Tianjin Medical University. A total of 12 SD rats were included. Predictor variableThe predictor variable was ZA exposure. Twelve SD rats were divided into two groups: experimental (n = 6) and control (n = 6), and they were intraperitoneally injected with ZA and saline, respectively. Main outcome variableThe outcome variables were histological and molecular changes. The maxilla, mandible, and ilium bone tissue samples were examined using Masson's trichrome and hematoxylin-eosin staining. Gene expression changes were identified using transcriptome sequencing, Kyoto encyclopedia of genes and genomes (KEGG), and gene interactome network analysis. The key changes were validated using the quantitative real-time polymerase chain reaction and immunohistochemistry. CovariatesNone. AnalysesThe t-test, Chi-square test, and Fisher's exact probability method were used for statistical analyses using the Statistical package for the social sciences software (version 26.0). ResultsAll animals remained healthy during the experiments. Histological staining revealed that the percentage of empty bone lacunae in the maxilla and mandible was significantly higher than that in the ilium (P < 0.01). In total, 552 genes were screened using transcriptome sequencing. The sonic hedgehog (Shh) signaling pathway was highly enriched. The key gene for the Shh interaction was distal-less homeobox 5 (Dlx-5). The Shh, Dlx-5, and bone morphogenetic protein 2 genes and protein expression levels in the maxilla and mandible were higher in the experimental group than in the control group (P < 0.05). Conclusions and RelevanceMRONJ-induced osteonecrosis and gene expression changes precede trauma-induced clinical changes in the SD rat model. These findings may provide additional support for timely and clinically early diagnosis and intervention.

Bone Marrow Morphology: A Key Diagnostic Tool in Various Hematological and Non Hematological Disorders

Bone marrow morphology means microscopic examination of bone marrow cells and tissue samples obtained by aspirate and trephine biopsy. An invaluable tool for diagnosis of many hematological and non-hematological disorders, various types of cancer staging and metastases detection. Different kinds of anemia and leukemia can be accurately diagnosed and monitored. Objective: To check the spectrum of diseases which could be identified and diagnosed with the help of bone marrow morphology examination thus demonstrating its diagnostic utility and to check diagnostic concordance between aspirate and core biopsy. Methods: Cross sectional study with consecutive sampling technique conducted over period of one year. Samples of both Bone Marrow Aspiration (BMA) and trephine biopsy (BMB) were collected from all patients included in study after consent using standard protocols. Air dried smears were prepared from bone marrow aspiration samples while trephine biopsy tissues were processed by histopathological techniques. Routine staining such as Wright Giemsa stain, H and E stain and special cytochemistry were used to visualize the cellular architecture of bone marrow. Results: Out of 471 samples, males were predominant with the frequency of 63.9% and maximum patients (31.8%) were from the age group of 35-40 years. Most common clinical indication to conduct bone marrow examination was unexplained Cytopenia which accounted for 35.2% cases, followed by suspicion of Leukemia (28.5%). Malignant hematological disorders were more common as compared to benign disorders (64% vs 17.5%). Acute leukemia was the most commonly identified cancer with frequency of 27.6%. Conclusions: Bone marrow morphology till date remains low risk, economical, crucial diagnostic tool in especially in under resource country like Pakistan. It can guide physicians to plan proper and timely management of patients.

Histological In Vivo Evaluation of Intense Pulsed Light Technology: Assessing the Safety on Oral Soft and Hard Tissues.

Three adult pigs were included in the trial. The oral cavity was divided into four quadrants and projected with a wide range of IPL settings. Alveolar bone, buccal mucosa, and gingival tissue samples were taken immediately and after 24 h. In each animal, one quadrant of the jaw was left untreated and served as a control. All samples were processed and stained with H&E. Clinical examination showed no evidence of changes in the integrity of the examined tissues. Histological examination of the different tissues did not demonstrate significant thermal damage or changes in the characterization of the cells compared to the control tissues. The use of IPL in the oral cavity is safe and does not negatively affect the tissues.

Integrating transcriptomic and proteomic data for a comprehensive molecular perspective on the association between sarcopenia and osteoporosis

BackgroundOsteoporosis and sarcopenia are common age-related conditions characterized by the progressive loss of bone density and muscle mass, respectively. Their co-occurrence, often referred to as osteosarcopenia, presents significant challenges in elderly care due to increased fragility and functional impairment. Existing studies have identified shared pathological mechanisms between these conditions, including inflammation, hormonal imbalances, and metabolic dysregulation, but a comprehensive understanding of their molecular interplay remains incomplete. ObjectiveThis study aims to deepen our understanding of the molecular interactions between sarcopenia and osteoporosis through an integrated omics approach, revealing potential therapeutic targets and biomarkers. MethodsEmploying a combination of proteomics and transcriptomics analyses, this study analyzed bone and muscle tissue samples from patients diagnosed with osteoporosis and osteosarcopenia. Techniques included high-throughput sequencing and label-free proteomics, supported by advanced bioinformatics tools for data analysis and functional annotation of genes and proteins. ResultsThe study found marked differences in gene and protein expressions between osteoporosis and osteosarcopenia tissues. Specifically, genes like PDIA5, TUBB1, and CYFIP2 in bone, along with MYH7 and NCAM1 in muscle, exhibited differential expression at both mRNA and protein levels. Pathway analyses revealed the significance of oxidative-reduction balance, cellular metabolism, and immune response in the progression of these conditions. Importantly, the study pinpointed osteoclast differentiation and NF-kappa B signaling pathways as critical in the molecular dynamics of osteosarcopenia, suggesting potential targets for therapy. ConclusionsThis study utilized transcriptomics and proteomics to identify key genes and proteins impacting sarcopenia and osteoporosis, employing advanced network tools to delineate interaction networks and crucial signaling pathways. It highlighted genes like PDIA5 and TUBB1, consistently expressed in both analyses, involved in pathways such as osteoclast differentiation and cytokine interactions. These insights enhance understanding of the molecular interplay in bone and muscle degeneration with aging, suggesting directions for future research into therapeutic interventions and prevention strategies for age-related degenerative diseases.

Methods to Enable Spatial Transcriptomics of Bone Tissues.

Understanding the relationship between the cells and their location within each tissue is critical to uncover the biological processes associated with normal development and disease pathology. Spatial transcriptomics is a powerful method that enables the analysis of the whole transcriptome within tissue samples, thus providing information about the cellular gene expression and the histological context in which the cells reside. While this method has been extensively utilized for many soft tissues, its application for the analyses of hard tissues such as bone has been challenging. The major challenge resides in the inability to preserve good quality RNA and tissue morphology while processing the hard tissue samples for sectioning. Therefore, a method is described here to process freshly obtained bone tissue samples to effectively generate spatial transcriptomics data. The method allows for the decalcification of the samples, granting successful tissue sections with preserved morphological details while avoiding RNA degradation. In addition, detailed guidelines are provided for samples that were previously paraffin-embedded, without demineralization, such assamples collected from tissue banks. Using these guidelines, high-quality spatial transcriptomics data generated from tissue bank samples of primary tumor and lung metastasis of bone osteosarcoma are shown.

Perbedaan Jumlah Hitung Osteoblas pada Pergerakan Ortodontik Gigi Setelah Pemberian Jintan Hitam

Osteoblasts are important for orthodontic tooth movement because they influence the activity of bone formation on the tension area. The administration of natural ingredients such as black cumin is proven to increase the number of osteoblasts because it contains thymoquinone as antioxidants to accelerate the process of tooth movement. The study aims to determine the differences in osteoblast count and their effect on the orthodontic movement of teeth after oral administration of black cumin. The method used in this research is experimental laboratory in vivo with a randomized post-test only control group design using 16 male Sprague-Dawley rat samples that were devided into four groups: the control group (K) with distilled water for 7 and 14 days, also the treatment group (P) with black cumin (Nigella sativa) for 7 and 14 days Elastomeric separators were placed on the upper jaw central incisors (right central incisor) using a separator applier to provide orthodontic mechanical force conditions. Alveolar bone tissue samples were taken after treatment then the number of osteoblasts was calculated histologically. Data were anaylzed using One Way Anova Test and LSD (p&lt;0.05). Result of this research indicated that there was an increase in the number of osteoblasts in orthodontic movement after oral administration of black cumin.

Targeting nicotinamide N-methyltransferase decreased aggressiveness of osteosarcoma cells.

Osteosarcoma (OS) is a primary bone malignancy that mostly affects young people, characterized by high metastatic potential, and a marked chemoresistance that is responsible for disease relapse in most patients. Therefore, it is necessary to identify novel molecules to setup targeted strategies to improve the clinical outcome. The enzyme nicotinamide N-methyltransferase (NNMT) catalyses the N-methylation of nicotinamide and other analogs, playing a crucial role in the biotransformation of drugs and xenobiotics. NNMT overexpression was reported in a wide variety of cancers, and several studies demonstrated that is able to promote cell proliferation, migration and resistance to chemotherapy. The aim of this study was to explore the potential involvement of NNMT in OS. Immunohistochemical analyses have been performed to evaluate NNMT expression in selected OS and healthy bone tissue samples. Subsequently, OS cell lines have been transfected with vectors targeting NNMT mRNA (shRNAs) and the impact of this downregulation on migration, cell proliferation, and response to chemotherapeutic treatment was also analysed by wound healing, MTT, SRB and Trypan blue assays, respectively. Results showed that OS samples display a significantly higher NNMT expression compared with healthy tissue. Preliminary results suggest that NNMT silencing in OS cell lines is associated to a decrease of cell proliferation and migration, as well as to enhanced sensitivity to chemotherapy. Data obtained showed that NNMT may represent an interesting marker for OS detection and a promising target for effective anti-cancer therapy.

Effective mechanical parameters of bone tissue samples for the selection of individual osteoimplants

Effective mechanical parameters of bone tissue samples for the selection of individual osteoimplants

Circadian rhythm-associated lncRNA RP11-414H17.5 as a key therapeutic target in osteosarcoma affects the tumor immune microenvironment and enhances malignancy

BackgroundIt has previously been proven that circadian rhythm disruption is associated with the incidence and deterioration of several tumors, which potentially leads to increased tumor susceptibility and a worse prognosis for tumor-bearing patients. However, their potential role in osteosarcoma has yet to be sufficiently investigated.MethodsTranscriptomic and clinical data of 84 osteosarcoma samples and 70 normal bone tissue samples were obtained from the TARGET and GTEx databases, circadian rhythm-related genes were obtained from Genecards, and circadian rhythm-related lncRNAs (CRLs) were obtained by Pearson correlation analysis, differential expression analysis, and protein–protein interaction (PPI) analysis. COX regression and LASSO regression were performed on the CRLs in order to construct a circadian rhythm-related prognostic prediction signature (CRPS). CRPS reliability was verified by Kaplan–Meier (KM), principal component analysis (PCA), nomogram, and receiver operating characteristic (ROC) curve. CRPS effects on the immune microenvironment of osteosarcoma were explored by enrichment analysis and immune infiltration analysis, and the effect of critical gene RP11-414H17.5 on osteosarcoma was experimentally verified.ResultCRPS consisting of three CRLs was constructed and its area under the curve (AUC) values predicted that osteosarcoma prognosis reached 0.892 in the training group and 0.843 in the test group, with a p value of < 0.05 for the KM curve and stable performance across different clinical subgroups. PCA analysis found that CRPS could significantly distinguish between different risk subgroups, and exhibited excellent performance in the prediction of the immune microenvironment. The experiment verified that RP11-414H17.5 can promote metastasis and inhibit apoptosis of osteosarcoma cells.ConclusionThe study revealed that circadian rhythm plays a crucial role in osteosarcoma progression and identified the impact of the key gene RP11-414H17.5 on osteosarcoma, which provides novel insights into osteosarcoma diagnosis and therapy.

Composition of pathogenic microorganism in chronic osteomyelitis based on metagenomic sequencing and its application value in etiological diagnosis

BackgroundTraditionally, conventional microbiological culture methods have been used to detect pathogenic microorganisms in chronic osteomyelitis. However, these methods have been found to have a low detection rate, complicating the precise guidance of infection treatment. This study employed metagenomic next-generation sequencing (mNGS) to detect these microorganisms in chronic osteomyelitis with three main objectives: 1). Gain a deeper understanding of the composition of pathogenic microorganisms in chronic osteomyelitis. 2). Compare the microbial detection rates between mNGS and the standard culture methods used in laboratories to enhance the effectiveness of the traditional culture methods. 3). Explore the potential of mNGS in etiological diagnosis.MethodsFifty clinically confirmed intraoperative bone tissue samples of chronic osteomyelitis from January 2021 to December 2021 were collected and subjected to mNGS and microbiological testing, respectively. The orthopaedic surgeon combined clinical manifestations and related examinations to determine the causative pathogens.ResultsThe culture method obtained 29 aerobic and parthenogenic anaerobic bacteria, 3 specific anaerobic bacteria, and 1 yeast-like fungus. Thirty-six aerobic and parthenogenic anaerobic bacteria, 11 specific anaerobic bacteria, and 1 yeast-like fungus were obtained by mNGS, and 2 Mycobacterium tuberculosis(MTB) strains were detected. However, there was no significant difference in the overall positive detection rate between mNGS and the culture method (P = 0.07), and the two were not statistically significant in detecting aerobic and partly anaerobic bacteria (P = 0.625). But, mNGS was significantly superior to culture in detecting anaerobic bacteria and Mycobacterium tuberculosis (P<0.05).ConclusionsThe mNGS method has enhanced our understanding of the distribution of pathogenic microorganisms in chronic osteomyelitis. Traditional culture methods help isolate and cultivate aerobic and facultative anaerobic bacteria, and fungi, and are also utilized for antibacterial drug sensitivity tests. However, mNGS has shown superior capabilities in detecting anaerobic bacteria, MTB, and mixed infection bacteria. This finding offers invaluable guidance for improving laboratory microbial culture and detection conditions. Hence, mNGS should be judiciously used for chronic osteomyelitis, and PCR can be implemented for certain difficult-to-culture microorganisms, such as MTB.

Inhibition of Caspase-1-mediated pyroptosis promotes osteogenic differentiation, offering a therapeutic target for osteoporosis

BackgroundPyroptosis, an emerging inflammatory form of cell death, has been previously demonstrated to stimulate a massive inflammatory response, thus hindering the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Nevertheless, the impact of pyroptosis in thwarting osteogenic differentiation and exacerbating the advancement of osteoporosis (OP) remains enigmatic. MethodsWe evaluated the expression levels of pyroptosis-associated indicators, including NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), CASPASE-1, IL-1β, and IL-18, in specimens obtained from femoral heads of OP patients, as well as in an ovariectomy-induced mouse model of OP. Subsequently, the precise roles of pyroptosis in osteogenic differentiation were investigated using bioinformatics analysis, alongside morphological and biochemical assessments. ResultsThe pivotal pyroptotic proteins, including NLRP3, Caspase-1, IL-1β, and IL-18, exhibited significant upregulation within the bone tissue samples of clinical OP cases, as well as in the femoral tissues of ovariectomy (OVX)-induced mouse OP model, displaying a negatively associated with compromised osteogenic capacity, as represented by lessened bone mass, suppressed expression of osteogenic proteins such as Runt-related transcription factor 2 (RUNX2), Alkaline phosphatase (ALP), Osterix (OSX), and Osteopontin (OPN), and increased lipid droplets. Moreover, bioinformatics analysis substantiated shared gene expression patterns between pyroptosis and OP pathology, encompassing NLRP3, Caspase-1, IL-1β, IL-18, etc. Furthermore, our in vitro investigation using ST2 cells revealed that dexamethasone treatment prominently induced pyroptosis while impeding osteogenic differentiation. Notably, gene silencing of Caspase-1 effectively counteracted the inhibitory effects of dexamethasone on osteogenic differentiation, as manifested by increased ALP activity and enhanced expression of RUNX2, ALP, OSX, and OPN. ConclusionOur findings unequivocally underscore that inhibition of Caspase-1-mediated pyroptosis promotes osteogenic differentiation, providing a promising therapeutic target for managing OP.

On Roth’s “human fossil” from Baradero, Buenos Aires Province, Argentina: morphological and genetic analysis

The “human fossil” from Baradero, Buenos Aires Province, Argentina, is a collection of skeleton parts first recovered by the paleontologist Santiago Roth and further studied by the anthropologist Rudolf Martin. By the end of the nineteenth century and beginning of the twentieth century it was considered one of the oldest human skeletons from South America's southern cone. Here, we present the results of an interdisciplinary approach to study and contextualize the ancient individual remains. We discuss the context of the finding by first compiling the available evidence associated with the historical information and any previous scientific publications on this individual. Then, we conducted an osteobiographical assessment, by which we evaluated the sex, age, and overall preservation of the skeleton based on morphological features. To obtain a 3D virtual reconstruction of the skull, we performed high resolution CT-scans on selected skull fragments and the mandible. This was followed by the extraction of bone tissue and tooth samples for radiocarbon and genetic analyses, which brought only limited results due to poor preservation and possible contamination. We estimate that the individual from Baradero is a middle-aged adult male. We conclude that the revision of foundational collections with current methodological tools brings new insights and clarifies long held assumptions on the significance of samples that were recovered when archaeology was not yet professionalized.

COL3A1, COL5A1 and COL6A2 serve as potential molecular biomarkers for osteoarthritis based on weighted gene co‑expression network analysis bioinformatics analysis.

Osteoarthritis (OA) is a non-inflammatory degenerative joint disease, characterized by joint pain and stiffness. The prevalence of OA increases with age. However, the relationship between biomarkers [collagen type III α1 (COL3A1), COL5A1, COL6A2, COL12A1] and OA remains unclear. The OA subchondral bone dataset GSE51588 was downloaded from the GEO database, and the differentially expressed genes (DEGs) were screened. Weighted gene co-expression network analysis was performed, and a protein-protein interaction network was constructed and further analyzed using Cytoscape and STRING. Functional enrichment analysis was performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and then Gene Set Enrichment Analysis (GSEA) was used to formulate the molecular functions and pathways based on the results of GO and KEGG analyses. Comparative Toxicogenomics Database and TargetScan were used to identify the hub-gene-related diseases and the microRNAs that regulated the central hub genes. Immunohistochemical staining was performed to confirm the expression of related proteins in OA and non-OA tissue samples. A total of 1,679 DEGs were identified. GO analysis showed that the DEGs were primarily enriched in the process of 'immune system', 'extracellular region', 'secretory granule', 'collagen-containing extracellular matrix', 'ECM-receptor, glycosaminoglycan binding' and 'systemic lupus erythematosus'. The results of GSEA were similar to those of GO and KEGG enrichment terms for DEGs. A total of 25 important modules were generated, and two core gene clusters and seven core genes were obtained (COL6A2, COL5A2, COL12A1, COL5A1, COL6A1, LUM and COL3A1). Core genes were expressed differentially between OA subchondral bone and normal tissue samples. The expression levels of COL3A1, COL5A1 and COL6A2 in OA subchondral bone tissue were higher compared with those in normal tissues, but COL12A1 expression was not significantly increased; all stained markers were highly expressed in surrounding tissues of immunohistochemical staining. In conclusion, COL3A1, COL5A1 and COL6A2 may be potential molecular biomarkers for OA.

The Influence of Combined Sterilization Factors on the Structural and Functional Characteristics of Bone Implants.

The results of a comprehensive study of the patterns of structural and functional changes in bone tissue samples after combined (ozone + radiation) sterilization are presented. The study used a different approach to the sterilization process with selective ozone or radiation exposure and an integral, combined one, based on a combined ozone-oxygen treatment of bone samples at the first stage and radiation at the second. The methods of IR spectroscopy, scanning electron microscopy with a prefix for elemental analysis, atomic force microscopy, and mechanical analysis with determination of elastic-plastic properties (Vickers microhardness index) were used in the work. It is shown that the ozone exposure used at the first stage of the combined sterilization process of bone implants does not lead to negative consequences with respect to their properties and characteristics. The results obtained serve as a scientific and methodological basis for the further improvement and optimization of sterilization technologies (including combined). They also offer a comprehensive justification of the parameters of sterilization regimes to ensure the safety of using bone implants during reconstructive operations, minimizing structural and functional changes in bone matter, and creating effective health-saving technologies and the possibility of using them for various biomedical applications.

Образ жизни детей и взрослых варварской округи Херсонеса (по материалам могильников Фронтовое 3 и Киль-Дере 1)

The necropolises Frontovoe 3 and Kil-Dere 1 from the environ of Chersonesos are interesting and important for the reconstruction of the cultural and historical situation on the borders of the Roman Empire. Data on the isotopic composition of bone collagen and tooth enamel provide new opportunities to discuss the nutrition, economy, and mobility of societies. The isotope composition of carbon and nitrogen informs about the average dietary intake of an individual over the last few years of life (7—10), while the ratio of strontium isotopes in tooth enamel characterizes the habitat of the child during the formation of the crown of the teeth (in this case, from 1 to 6 years). Thus, we obtain information about local natives and migrants of the first generation, whose childhood, probably, passed outside the environ of Chersonesos. The isotopic composition of carbon and nitrogen in 67 samples of bone tissue and the composition of strontium in 35 samples of tooth enamel were studied. The life support system of these societies was based on the use of local terrestrial food resources. Few individuals have been found who can be named first-generation migrants. The low variability of the isotope ratios of strontium and carbon indicates a clear localization of this population group on the given territory. We also discuss the factors behind the differences in the isotopic markers of groups from Kil-Dere 1 and Frontovoye 3. These can be both environmental factors and cultural differences. The questions addressed here can be answered by studying a broader base of the compared materials.

Identification and validation of long noncoding RNA AC083900.1 and RP11-283C24.1 for prediction of progression of osteosarcoma

BackgroundThe role of cuproptosis, an emerging cell death pathway that makes a remarkable contribution to tumor progression, remains elusive in osteosarcoma (OS), in addition to its regulator, including long-no-coding RNAs (lncRNAs) that are also a critical factor for fueling OS. MethodsTranscriptome and clinical data from 70 normal human bone tissue samples and 84 frozen clinical osteosarcoma samples were included in this study. Cuproptosis-associated lncRNAs (CRlncs) were identified through differential expression and co-expression analyses. Univariate Cox regression was performed to screen for prognostic lncRNAs, then we used least absolute shrinkage and selection operator regression to distinguish prognosis-related CRlncs (AC083900.1 and RP11-283C24.1) for modeling the CRlncs prognostic signature (CLPS) by multivariate Cox regression using the stepwise method. CLPS performance was tested by independent prognostic analyses, survival curve and receiver operating characteristic (ROC) curve. In addition, the molecular and immune mechanisms that underlie the unfavorable prognosis of CLPS-identified high-risk group were elucidated. ResultAC083900.1 and RP11–283C24.1 have been identified as the most important CRlncs for OS progression (hazard ratio: 3.498 and 2.724, respectively), and the derived CLPS demonstrated outstanding performance for the prediction of OS prognosis (AUC of 0.799 and 0.778 in the training and test sets, both adj-p < 0.05 in survival curve). As was anticipated, CLPS also outperformed a recent clinical prognostic approach that only achieved an AUC of 0.682 [metastasis]. It is notable that AC083900.1 progressed OS metastasis, evidenced by its high expression in metastatic OS, its high correlation to metastasis-related genes, and its high AUC of 0.683 for the prediction of metastasis. Mechanistically, AC083900.1 and RP11–283C24.1 dysregulated many critical biological processes regarding humoral immune response, immunoglobulin complex, etc.; while reducing the infiltration of many cytotoxic immune cells (B-cells, TIL, neutrophils, etc.). It is encouraging that BMS-509744 and KIN001–135 demonstrated high therapeutic implications for CLPS-identified high-risk OS, and the low-risk counterpart was sensitive to SB-216763. Quantitative RT-PCR analysis showed that both AC083900.1 and RP11-283C24.1 were significantly upregulated in different osteosarcoma cell lines. ConclusionThis study elucidated the roles and mechanisms of AC083900.1 and RP11-283C24.1 in the development of OS, fostering a reliable prognostic approach and treatment for OS patients.

Lycopene enhances bone neoformation in calvaria bone defects of ovariectomized rats.

Osteoporosis can affect a significant part of the population and fractures are the most common complications associated with this disease, leading to high public health costs. Thus, the prevention of fractures is relevant to individuals with signs and symptoms as well as to the health system. Postmenopausal osteoporosis has been associated with oxidative stress, emphasizing the importance of an efficient defense system to maintain bone health. Lycopene is a carotenoid with antioxidant properties that may stimulate osteoblastogenesis and inhibit osteoclastogenesis. The purpose of this investigation was to analyze the influence of lycopene in the bone neoformation of calvaria defects in ovariectomized rats utilizing the concentration of 45 mg/kg. Wistar Hannover female rats were divided into ovariectomized and sham groups. The ovariectomized animals received 45 mg/kg lycopene (OvxL) or water (Ovx) by daily gavage the day after ovariectomy/sham surgery for 16 weeks. Twelve weeks after ovariectomy, there were performed 5-mm calvaria defects followed by euthanasia after 4 weeks. Samples of bone tissue were collected to perform morphological and morphometrical analysis of the neoformed bone area, and percentage with Software Image J. Morphological evaluation showed mature bone with more osteocytes in the group OVxL when compared to the other groups. The morphometrical analysis demonstrated a significant increase of bone neoformation in the group OvxL (p<0.05). The data obtained suggest that lycopene benefits bone repair in the absence of estrogenic hormones.

Influence of Er:YAG laser irradiation on the outcomes of alveolar ridge preservation at the infected molar sites: a randomized controlled trial

BackgroundThe purpose of this study was to investigate the socket healing outcome after alveolar ridge preservation at infected molar sites using an erbium-doped yttrium aluminium garnet (Er:YAG) laser.MethodsEighteen patients who needed molar extraction and exhibited signs of infection were included and allocated into either the laser group or the control group. Er:YAG laser irradiation for degranulation and disinfection was performed with alveolar ridge preservation (ARP) in the laser group. Traditional debridement with a curette was performed in the control group. Two months after ARP, bone tissue samples were harvested at the time of implant placement for histological analysis. Assessment of dimension changes in alveolar bone was conducted by superimposing two cone-beam computed tomography (CBCT) scans taken at baseline and two months after extraction.ResultsHistologically, after two months of healing, Er:YAG laser treatment resulted in more newly formed bone (laser: 17.75 ± 8.75, control: 12.52 ± 4.99, p = 0.232). Moreover, greater osteocalcin (OCN) positive expression and lower runt-related transcription factor 2 (RUNX-2) positive expression were detected in the laser group. However, no statistically significant difference was observed between the two groups. The difference in the vertical resorption of the buccal bone plate was statistically significant between groups (laser: -0.31 ± 0.26 mm, control: -0.97 ± 0.32 mm, p < 0.05). Major changes in ridge width were observed at 1 mm below the bone crest. However, the differences between groups were not significant (laser: -0.36 ± 0.31 mm, control: -1.14 ± 1.24 mm, p = 0.171).ConclusionsARP with Er:YAG laser irradiation seemed to improve bone healing by regulating osteogenesis-related factor expression in the early stage at infected sites.Trial registrationThe trial was registered on the Chinese Clinical Trial Registry Platform (https://www.chictr.org.cn/) (registration number: ChiCTR2300068671; registration date: 27/02/2023).

Temperature Monitoring of Two Different Thermal Boxes for the Transport of Biological Samples

According to the Resolutions of the Collegiate Board of Directors RDC No. 20/2014, 214/2018, and 707/2022, validation of the temperature of thermal boxes for the transport of biological samples must be based on standardized procedures and tested by the Tissue Banks, guaranteeing safety and quality. Therefore, they can be simulated. Our objective was to monitor and compare the temperature of 2 different coolers while transporting biological samples. The following items were packed in each of the 2 different thermal boxes (Box 1: Easy Path; Box 2: Safe Box Polyurethane Vegetal): 6 blood samples (30 mL), one bone tissue sample (200 g), 8 hard ice (Gelox, to keep the temperature <8ºC), and internal and external traceability "Time Stamp" sensors installed for measuring and storing temperature data in real-time. The monitored boxes were placed in the trunk of a bus that traveled an approximate distance of 630 km and were then placed in the trunk of a car, under direct sunlight, until they reached a temperature of 8ºC. In Box 1, the internal temperature was maintained in the range of -7ºC to 8ºC for approximately 26 hours. In Box 2, the internal temperature was maintained in the range of -10ºC to 8ºC for approximately 98 hours and 40 minutes. We concluded that both coolers, under similar storage conditions, are suitable for transporting biological samples, with Box 2 maintaining the desired temperature for longer.

Circ_0006873 suppresses the osteogenic differentiation of human‐derived mesenchymal stem cells through mediating miR‐20a/SMURF2 axis in vitro

The clinical application of human-derived mesenchymal stem cells (hMSCs) in osteoporosis (OP) treatment is promising. We aimed to uncover the role of circular RNA 0006873 (circ_0006873) in OP progression using hMSCs. The levels of circ_0006873, pantothenate kinase 2 (PANK2) messenger RNA (mRNA), microRNA-20a (miR-20a), SMAD specific E3 ubiquitin protein ligase 2 (SMURF2) mRNA and the mRNA levels of osteogenesis-related markers were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The protein expression of osteogenesis-related markers and SMURF2 was detected by Western blot assay. Alkaline phosphatase (ALP) staining and activity were determined using an ALP staining Kit and an ALP Colorimetric Assay Kit. Circ_0006873 was highly expressed in the serum samples and bone tissue samples of OP patients compared with control cases. Circ_0006873 overexpression down-regulated the expression of osteogenesis-related markers and reduced ALP staining and activity. Circ_0006873 down-regulated miR-20a level through its interaction with miR-20a in hMSCs. Circ_0006873 suppressed osteogenic differentiation through targeting miR-20a. SMURF2 was a molecular target of miR-20a, and miR-20a promoted osteogenic differentiation through targeting SMURF2. Circ_0006873 suppressed the osteogenic differentiation of hMSCs by upregulating SMURF2 level via sponging miR-20a in vitro.