Human Bone Research Articles

A model of long-term population growth with an application to Central West Argentina.

We propose an Ideal Specialization Model to help explain the diversity of population growth trajectories exhibited across archaeological regions over thousands of years. The model provides a general set of expectations useful for guiding empirical research, and we provide a concrete example by conducting a preliminary evaluation of three expectations in Central West Argentina. We use kernel density estimates of archaeological radiocarbon, estimates of paleoclimate, and human bone stable isotopes from archaeological remains to evaluate three expectations drawn from the model's dynamics. Based on our results, we suggest that innovations in the production of food and social organization drove demographic transitions and population expansion in the region. The consistency of population expansion in the region positively associates with changes in diet and, potentially, innovations in settlement and social integration.

Activation of Mitochondria in Mesenchymal Stem Cells by Mitochondrial Delivery of Coenzyme Q10.

The efficacy of mesenchymal stem cell (MSC) transplantation has been reported for various diseases. We previously developed a drug delivery system targeting mitochondria (MITO-Porter) by using a microfluidic device to encapsulate Coenzyme Q10 (CoQ10) on a large scale. The current study aimed to confirm if treatment with CoQ10 encapsulated by MITO-Porter enhanced mitochondrial functions in MSCs, with the potential to improve MSC transplantation therapy. We used highly purified human bone marrow-derived MSCs, described as rapidly expanding clones (RECs), and attempted to control and increase the amount of CoQ10 encapsulated in the MITO-Porter using microfluidic device system. We treated these RECs with CoQ10 encapsulated MITO-Porter, and evaluated its cellular uptake, co-localization with mitochondria, changes in mitochondrial respiratory capacity, and cellular toxicity. There was no significant change in mitochondrial respiratory capacity following treatment with the previous CoQ10 encapsulated MITO-Porter; however, mitochondrial respiratory capacity in RECs was significantly increased by treatment with CoQ10-rich MITO-Porter. Utilization of a microfluidic device enabled the amount of CoQ10 encapsulated in MITO-Porter to be controlled, and treatment with CoQ10-rich MITO-Porter successfully activated mitochondrial functions in MSCs. The MITO-Porter system thus provides a promising tool to improve MSC cell transplantation therapy.

Pro-Osteogenic Effect of the Nutraceutical BlastiMin Complex® in Women with Osteoporosis or Osteopenia: An Open Intervention Clinical Trial

Osteoporosis is a chronic disease that affects millions of patients worldwide and is characterized by low bone mineral density (BMD) and increased risk of fractures. Notably, natural molecules can increase BMD and exert pro-osteogenic effects. Noteworthily, the nutraceutical BlastiMin Complex® (Mivell, Italy, European Patent Application EP4205733A1) can induce differentiation of human bone marrow mesenchymal stem cells (BM-MSCs) in osteoblasts and can exert in vitro pro-osteogenic and anti-inflammatory effects. Thus, the purpose of this study was to verify the effects of BlastiMin Complex® on bone turnover markers (BTMs) and BMD in patients with senile and postmenopausal osteopenia or osteoporosis. The efficacy of BlastiMin Complex® on BTMs in serum was evaluated through biochemical assays. BMD values were analyzed by dual-energy X-ray absorptiometry (DXA) and Radiofrequency Echographic Multi Spectrometry (R.E.M.S.) techniques, and the SNPs with a role in osteoporosis development were evaluated by PCR. Clinical data obtained after 12 months of treatment showed an increase in bone turnover index, a decrease in C-reactive protein levels, and a remarkable increase in P1NP levels, indicating the induction of osteoblast proliferation and activity in the cohort of 100% female patients recruited for the study. These findings show that the nutraceutical BlastiMin Complex® could be used as an adjuvant in combination with synthetic drugs for the treatment of osteoporosis pathology.

Autogenous Bilateral Cleft Alveolar Bone Grafting with rhBMP-2 and Cellular Bone Matrix.

Alveolar bone grafting (ABG) in bilateral cleft lip and palate (BCLP) patients provides a reconstructive challenge. We present a novel technique of combining autologous iliac crest bone graft (ICBG) with recombinant human bone morphogenic protein 2 (rhBMP-2) and cellular bone matrix (CBM) for ABG in BCLP patients. Complete bone fill occurred in 90% of patients, with 100% having bilateral canine eruption. No patients required repeat ABG, and no significant complications were reported. The alveolar cleft gap volume significantly decreased with an improvement of 75.87%. ABG with autologous ICBG with rhBMP-2 and CBM is an effective technique for patients with BCLP.

Elastoplastic and Electrochemical Characterization of xTiB2 Strengthened Ti Porous Composites for Their Potential Biomedical Applications

The microstructure, elastoplastic properties, and corrosive response of induced porous Ti-TiH2 materials reinforced with TiB2 particles were investigated. Samples were fabricated using CP-Ti Grade1, Titanium Hydride (TiH2), TiB2 powders (0, 3, 10, and 30 vol.%), and ammonium bicarbonate salt (40 vol.%) as a space holder. Composites were fabricated using the Powder Metallurgy technique under high-vacuum conditions (HVS) at 1100 °C. Scanning electron microscopy, X-ray diffraction, nanoindentation tests, and electrochemical assays were used to investigate the pore formation, pore distribution, phase formation, elastoplastic properties, and electrochemical behavior of the compounds, respectively. With a mean pore diameter of 50–900 µm and Young’s modulus of less than 100 GPa, which is close to the properties of human bone, the pore structures of the compounds processed here are shown to be a potential biomaterial for osseointegration. In addition, their H/Er and H3/Er2 ratios for the reinforced samples are higher than those of the unreinforced sample (1.5 and 4 times higher than the unreinforced sample, respectively), suggesting a better wear resistance of the Ti-TiH2/xTiB2 composites. Electrochemical experiments demonstrated that the Ti-TiH2/xTiB2 composites exhibited superior passivation properties compared to the Ti-TiH2 sample. Additionally, the corrosion rates exhibited by the 3 and 10 vol.% of TiB2 samples were found to be within an acceptable range for potential biomedical applications (29.26 and 185.82 E-3 mm·y−1). The elastoplastic properties combined with the electrochemical behavior place the Ti-TiH2/3-10TiB2 composites as potential candidates for the biomedical application of CP-Ti.

Investigation of the Relationship between Surface Features, Protein Adsorption, and Osteoimmunomodulation: The Case of a Chemically Treated Titanium Alloy.

This paper deals with the combined effects of immune response and osseointegration because of the lack of comprehensive studies on this topic. An antibacterial Ti surface was considered because of the high risk of infection for titanium bone implants. A chemically treated Ti6Al4 V alloy [Ti64(Sr-Ag)] with a microporous and Sr-Ag doped surface was compared to a polished version (Ti64) regarding protein adsorption (albumin and fibronectin) and osteoimmunomodulation. Characterization via fluorescence microscopy and zeta potential showed a continuous fibronectin layer on Ti64(Sr-Ag), even with preadsorbed albumin, while it remained filamentous on Ti64. Macrophages (differentiated from THP-1 monocytes) were cultured on both surfaces, with viability and cytokine release analyzed. Differently from Ti64, Ti64(Sr-Ag) promoted early anti-inflammatory responses and significant downregulation of VEGF. Ti64(Sr-Ag) also enhanced human bone marrow mesenchymal cell differentiation toward osteoblasts, when a macrophage-conditioned medium was used, influencing ALP production. Surface properties in relation to protein adsorption and osteoimmunomodulation were discussed.

Three-dimensional-printed femoral diaphysis for biomechanical testing-Optimization and validation.

Polylactic acid (PLA) models of normal human femoral diaphyses were designed using three-dimensional (3D) printing technology to create inexpensive, accessible, and reproducible specimens for flexural biomechanical studies. These models were subjected to three-point bending and their response to loading was characterized. The anisotropic mechanical behavior of the 3D-printed femurs and the influence of printing orientations, infill density, wall layers, resolution, and other printing parameters were explored to develop a design space. The objective of the design space was set to emulate the flexural biomechanical response of the normal human femur bones. Results show the 3D-printed PLA diaphyseal femurs with 5% infill density, two-four wall layers, and a resolution of 200 µm resulted in a flexural strength of 184.8 ± 8.18 MPa. Models with 20% infill density and six wall layers resulted in a flexural modulus of 18.54 ± 0.543 GPa. These results emulate the biomechanical response of the normal human femur, as determined by historical target values derived from prior cadaveric and 3D printing data. With further research, inclusive of modeling the proximal and distal femur and more comprehensive biomechanical testing, 3D-printed femurs may ultimately serve as a cheap, accessible biomechanical resource for surgeons and researchers.

Integration of animal husbandry and millet agriculture in Bronze Age East-central Eurasia revealed by faunal stable isotopes at the Jirentai Goukou site, Xinjiang

How food globalization in the Bronze Age influenced the economies and societies of regions along the Eurasian continent has attracted widespread attention. As an integral part of this food globalization, previous studies have focused on the impact of the westwards spread of millet crops on the human populations of East-Central Eurasia. However, the relationship between millet crops and key subsistence economies, such as animal husbandry, which humans rely on in East-Central Eurasia, remains unclear. Here, we carry out a new stable isotope study of 68 domesticated faunal bones from Bronze Age Jirentai Goukou site, Xinjiang, and combine about 1500 published stable isotope data of human and faunal bones to explore systemically the relationship between animal husbandry and millet agriculture in Bronze Age East-Central Eurasia. Our study reveals first the integration of animal husbandry and millet agriculture in Bronze Age Xinjiang, which is indicated by the adoption of C4 millet byproducts to feed domesticated sheep/goats and cattle. Further wide-range stable isotopic analysis in Bronze Age East-Central Eurasia highlights the existence of two different patterns in the relationship between animal husbandry and millet agriculture: the Central Plains pattern and the Inner Asia pattern. These differences can be expressed clearly in several aspects, including the species of domesticated herbivores fed, the degree, method and seasonality of feeding sheep/goats and cattle, and the composition of fodder. On this basis, the possible reasons for the different patterns in the relationship between millet agriculture and animal husbandry in East-Central Eurasia are further explored, such as the different histories of agricultural and pastoral development between the Central Plains and Inner Asia, the distinct characteristics of agriculture and pastoralism (sedentary and mobile), and the specific social, economic/political and geographical environments. Our study provides important information for understanding the specific impact of the globalization of millet agriculture on the animal husbandry economy in Bronze Age Eurasia.

Neuroendocrinology of bone.

The past decade has witnessed significant advances in our understanding of skeletal homeostasis and the mechanisms that mediate the loss of bone in primary and secondary osteoporosis. Recent breakthroughs have primarily emerged from identifying disease-causing mutations and phenocopying human bone disease in rodents. Notably, using genetically-modified rodent models, disrupting the reciprocal relationship with tropic pituitary hormone and effector hormones, we have learned that pituitary hormones have independent roles in skeletal physiology, beyond their effects exerted through target endocrine glands. The rise of follicle-stimulating hormone (FSH) in the late perimenopause may account, at least in part, for the rapid bone loss when estrogen is normal, while low thyroid-stimulating hormone (TSH) levels may contribute to the bone loss in thyrotoxicosis. Admittedly speculative, suppressed levels of adrenocorticotropic hormone (ACTH) may directly exacerbate bone loss in the setting of glucocorticoid-induced osteoporosis. Furthermore, beyond their established roles in reproduction and lactation, oxytocin and prolactin may affect intergenerational calcium transfer and therefore fetal skeletal mineralization, whereas elevated vasopressin levels in chronic hyponatremic states may increase the risk of bone loss.. Here, we discuss the interaction of each pituitary hormone in relation to its role in bone physiology and pathophysiology.

Cheminformatic Read-Across Approach Revealed Ultraviolet Filter Cinoxate as an Obesogenic Peroxisome Proliferator-Activated Receptor γ Agonist.

This study introduces a novel cheminformatic read-across approach designed to identify potential environmental obesogens, substances capable of disrupting metabolism and inducing obesity by mainly influencing nuclear hormone receptors (NRs). Leveraging real-valued two-dimensional features derived from chemical fingerprints of 8435 Tox21 compounds, cluster analysis and subsequent statistical testing revealed 385 clusters enriched with compounds associated with specific NR targets. Notably, one cluster exhibited selective enrichment in peroxisome proliferator-activated receptor γ (PPARγ) agonist activity, prominently featuring methoxy cinnamate ultraviolet (UV) filters and obesogen-related compounds. Experimental validation confirmed that 2-ethoxyethyl 4-methoxycinnamate, an organic UV filter cinoxate, could selectively bind to PPARγ (Ki = 18.0 μM), eliciting an obesogenic phenotype in human bone marrow-derived mesenchymal stem cells during adipogenic differentiation. Molecular docking and further experiments identified cinoxate as a potent PPARγ full agonist, demonstrating a preference for coactivator SRC3 recruitment. Moreover, cinoxate upregulated transcription levels of genes encoding lipid metabolic enzymes in normal human epidermal keratinocytes as primary cells exposed during clinical usage. This study provides compelling evidence for the efficacy of cheminformatic read-across analysis in prioritizing potential obesogens, showcasing its utility in unveiling cinoxate as an obesogenic PPARγ agonist.

Small Nuclear Ribonucleoprotein Polypeptides B and B1 Promote Osteosarcoma Progression via Activating the ATM Signaling Pathway through Ribonucleotide Reductase Subunit M2

Osteosarcoma is a malignant bone tumor characterized by high metastatic potential and recurrence rates post-therapy. The small nuclear ribonucleoprotein polypeptides B and B1 (SNRPB), a core component of spliceosome, have been reported to exhibit upregulation across several cancer types. However, the precise role of SNRPB in osteosarcoma progression remains poorly elucidated. Herein, we explored SNRPB expression in human osteosarcoma tissues and normal bone tissues by immunohistochemical staining (IHC) staining, revealing a notable upregulation of SNRPB in osteosarcoma, correlating with diminished survival rates. Moreover, the in vitro loss-of-function experiments showed that SNRPB knockdown significantly suppressed the osteosarcoma cell proliferation and migration, as well as tubule formation of HUVECs, while enhancing osteosarcoma cell apoptosis. Mechanistically, we revealed that SNRPB promoted the transcription of ribonucleotide reductase subunit M2 (RRM2) via E2F transcription factor 1 (E2F1). Further rescue experiments indicated that RRM2 was required for SNRPB-induced malignant behaviors in osteosarcoma. Additionally, we confirmed that the function of SNRPB in osteosarcoma cell growth and apoptosis was associated with ATM signaling pathway activation. In conclusion, our findings provide initial insights into the underlying mechanisms governing SNRPB-induced osteosarcoma progression and proposed for SNRPB as a novel therapeutic target in osteosarcoma management.

Development of a continuum-based, meshless, finite element modeling approach for representation of trabecular bone indentation

Implant subsidence into the underlying trabecular bone is a common problem in orthopaedic surgeries; however, the ability to pre-operatively predict implant subsidence remains limited. Current state-of-the-art computational models for predicting subsidence have issues addressing this clinical problem, often resulting from the size and complexity of existing subject-specific, image-based finite element (FE) models. The current study aimed to develop a simplified approach to FE modeling of subject-specific trabecular bone indentation resulting from implant penetration.Confined indentation experiments of human trabecular bone with flat- and sharp-tip indenters were simulated using FE analysis. A generalized continuum-level approach using a meshless smoothed particle hydrodynamics (SPH) approach and an isotropic crushable foam (CF) material model was developed for the trabecular bone specimens. Five FE models were generated with CF material parameters calibrated to cadaveric specimens spanning a range of bone mineral densities (BMD). Additionally, an alternative model configuration was developed that included consideration of bone marrow, with bone and marrow material parameters assigned to elements randomly according to bone volume (BV%) measurements of experimental specimens, owing to the non-uniform nature of trabecular bone tissue microstructure.Statistical analysis found significant correlation between the shapes of the numerical and experimental force-displacement curves. FE models accurately captured the bone densification patterns observed experimentally. Inclusion of marrow elements offered improved response prediction of the flat-tip indenter tests.Ultimately, the developed approach demonstrates the ability of a generalizable continuum-level SPH approach to capture bone variability using clinical bone imaging metrics without needing detailed image-based geometries, a significant step towards simplified subject-specific modeling of implant subsidence.

Expression of TGFβ-1 and CTGF in the Implanted Cochlea and its Implication on New Tissue Formation.

Transforming growth factor beta-1 (TGFβ-1) and connective tissue growth factor (CTGF) are upregulated in the implanted human cochlea. Cochlear implantation can lead to insertion trauma and intracochlear new tissue formation, which can detrimentally affect implant performance. TGFβ-1 and CTGF are profibrotic proteins implicated in various pathologic conditions, but little is known about their role in the cochlea. The present study aimed to characterize the expression of these proteins in the human implanted cochlea. Archival human temporal bones (HTB) acquired from 12 patients with previous CI and histopathological evidence of new tissue formation as well as surgical samples of human intracochlear scar tissue surrounding the explanted CI were used in this study. Histopathologic analysis of fibrosis and osteoneogenesis was conducted using H&E. Protein expression was characterized using immunofluorescence. RNA expression from surgical specimens of fibrotic tissue surrounding the CI was quantified using qRT-PCR. TGFβ-1 and CTGF protein expressions were upregulated in the areas of fibrosis and osteoneogenesis surrounding the CI HTB. Similarly, surgical samples demonstrated upregulation of protein and mRNA expression of TGFβ-1 and mild upregulation of CTGF compared with control. TGFβ-1 was expressed diffusely within the fibrous capsule, whereas CTGF was expressed in the thickened portion toward the modiolus and the fibrosis-osteoneogensis junction. To our knowledge, this is the first study to demonstrate increased expression of TGFβ-1 and CTGF in the human implanted cochlea and may provide better understanding of the mechanism behind this pathogenic process to better develop future mitigating interventions.

Development of a composite using calcined bone powder and silane cross-linked alginate as bone substitute material.

Calcined bone is an attractive natural material for use as a bone substitute because of its cost-effectiveness and high biocompatibility, which are comparable to that of synthetic hydroxyapatite. However, the calcination process has significantly weakened the mechanical properties. In this study, a composite of calcined bovine bone powder reinforced with silane cross-linked alginate was prepared to assess its biocompatibility, osteoconductivity, and mechanical compatibility as a bone substitute material. Culture studies with osteoblast-like cells (MC3T3-E1) showed no cytotoxicity toward the composite and exhibited general cell proliferative properties in its presence. In contrast, the composite reduced the alkaline phosphatase activity of osteoblasts but led to significant noncellular apatite deposition on the surface. In addition, quasi-static compression tests of the composite revealed mechanical properties comparable to those of human cancellous bone. The mechanical properties remained stable under wet conditions and did not deteriorate significantly even after 2 weeks of immersion in simulated body fluid at 37°C. The results show that this composite, composed of calcined bone powder and silane cross-linked alginate, is a promising bone substitute material with biocompatibility, osteoconductivity, and mechanical compatibility.

Three-dimensional replica of the temporal bone in the teaching of human anatomy.

The current study proposes the comparison of the visualization and identification of anatomical details between natural human temporal bone, its respective copy from three-dimensional printing, and the virtual model obtained from CBCT. The sample consisted of undergraduate students in Dentistry (Group UE, n = 22), Postgraduate students in Radiology and Imaging (Group P-RI, n = 20), and Postgraduate students in Forensic Odontology (Group P-FO, n = 24). All participants attended a theoretical class on specialized anatomy of the temporal bone and subsequently performed the markings of 10 determined structures. The number of correct identifications was similar in natural bone and printed three-dimensional models in all groups (p > 0.05). The virtual model showed a significantly lower number of correct structures (p < 0.05) in the 3 groups. In general, there were significantly higher percentages of accurate answers among postgraduate students compared to undergraduate students. Most graduate students believed that the printed three-dimensional model could be used to teach anatomy in place of natural bone, while undergraduate students disagreed or were unsure (p < 0.05). Regarding the virtual tomographic image, in all groups, students disagreed or were not sure that its use would be beneficial in replacing natural bone. Three-dimensional and virtual models can be used as auxiliary tools in teaching anatomy, complementing practical learning with natural bones.

Oxaliplatin-induced upregulation of exosomal miR-424-3p derived from human bone marrow mesenchymal stem cells attenuates progression of gastric cancer cells

Chemotherapy, particularly with oxaliplatin, is a key treatment for advanced gastric cancer (GC), and exosomes derived from human bone marrow mesenchymal stem cells (hBM-MSCs) play a vital role in the tumor microenvironment. The study aims to elucidate the previously unexplored role of exosomes derived from hBM-MSCs in GC tumorigenesis, especially under the influence of chemotherapy. We conducted an experimental study, utilizing miRNA sequencing and biological experiments, to analyze the tumorigenicity of exosomal miR-424-3p secreted by hBM-MSCs and its target gene RHOXF2 in GC cell lines. The results were confirmed through experimentation using a xenograft mouse model. This study demonstrated the role of hBM-MSCs in the GC microenvironment, focusing on their epithelial-mesenchymal transition (EMT) facilitation through exosomes, which led to enhanced tumorigenicity in GC cells. Intriguingly, this pro-tumor effect was abrogated when hBM-MSCs were treated with oxaliplatin. Exosomal miRNA sequencing revealed that oxaliplatin can upregulate the levels of miR-424-3p in exosomes secreted by hBM-MSCs, thereby inhibiting the EMT process in GC cells. Furthermore, miR-424-3p was identified to target and downregulate RHOXF2 expression, impeding the malignant behavior of GC cells both in vitro and in the mouse model. These findings uncover a potential hidden mechanism of oxaliplatin's anti-tumor action and propose the delivery of miR-424-3p via exosomes as a promising avenue for anti-tumor therapy.

CBP-mediated FOXO4 acetylation facilitates postmenopausal osteoporosis (PMO) progression through the inhibition of the Wnt/β-catenin signaling pathway.

FOXO4 was previously identified as a potential biomarker and therapeutic target for postmenopausal osteoporosis (PMO) using bioinformatic analysis, but its specific function and molecular mechanism in the progression of osteoporosis was not reported. The current study was designed to investigate the biological function and underlying mechanism of FOXO4 in PMO. Our results showed that FOXO4 expression was significantly upregulated in the serum samples of PMO patients, which was also negatively correlated with the expression of osteogenesis genes (OCN and ALP). In addition, FOXO4 depletion alleviated osteoporosis by facilitating osteogenic differentiation and inhibiting adipogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs). Overexpression of FOXO4 exerted the opposite effects on the osteogenic/adipogenic differentiation in hBMSCs. Moreover, FOXO4 knockdown activated the Wnt/β-catenin signaling whereas the inhibition of Wnt/β-catenin signaling overturned the effects of FOXO4 deficiency on osteoporosis. Furthermore, FOXO4 upregulation in PMO was caused by CBP-induced acetylation. In summary, our data demonstrated that FOXO4 was a potent biomarker for PMO and mediated the balance between osteogenesis and adipogenesis in hBMSCs by regulating Wnt/β-catenin signaling.

Analysis of the milling response of an artificial temporal bone developed for otologic surgery in comparison with human cadaveric samples

Temporal-bone milling is a delicate process commonly performed during otologic surgery to gain access to the middle and inner ear structures. Because of the numerous at-risk structures of this anatomic area, extensive surgeon training is required. Artificial temporal bones offer an interesting alternative to cadaveric training. However, the evaluation of such simulators has not been systematic, with an absence of objective validation of their milling response, especially in a surgical context.By measuring the milling forces obtained during the classical steps of otologic surgery on six 3D-printed and three cadaveric temporal bones, this work aims at evaluating the ability of the OTOtwin® synthetic temporal bone to reproduce human bone behavior.A better repeatability was obtained for artificial bones than for cadaveric ones. However, the level of forces recorded during artificial bone milling was close to the one measured wIth cadaveric samples. The effect of both surgical phase and irrigation on milling force levels were also quantified. The experiments conducted in this study confirmed the suitability of OTOtwin® temporal bone model for both otologic surgery training and research purposes. Valuable insights were also gained from this study regarding the understanding of the otologic milling process.

Biosynthesized Silver Nanoparticles from Cyperus conglomeratus Root Extract Inhibit Osteogenic Differentiation of Immortalized Mesenchymal Stromal Cells.

Silver nanoparticles (AgNPs) are a focus of huge interest in biological research, including stem cell research. AgNPs synthesized using Cyperus conglomeratus root extract have been previously reported but their effects on mesenchymal stromal cells have yet to be investigated. The aim of this study is to investigate the effects of C. conglomeratus-derived AgNPs on adipogenesis and osteogenesis of mesenchymal stromal cells. AgNPs were synthesized using C. conglomeratus root extract, and the phytochemicals involved in AgNPs synthesis were analyzed using gas chromatography-mass spectrometry (GCMS). The cytotoxicity of the AgNPs was tested on telomerase-transformed immortalized human bone marrow-derived MSCs-hTERT (iMSC3) and human osteosarcoma cell line (MG-63) using MTT and apoptosis assays. The uptake of AgNPs by both cells was confirmed using inductively coupled plasma-optical emission spectrometry (ICP-OES). Furthermore, the effect of AgNPs on iMSC3 adipogenesis and osteogenesis was analyzed using stain quantification and reverse transcription- quantitative polymerase chain reaction (RT-qPCR). The phytochemicals predominately identified in both the AgNPs and C. conglomeratus root extract were carbohydrates. The AgNP concentrations tested using MTT and apoptosis assays (0.5-64 µg/ml and 1,4 and 32 µg/ml, respectively) showed no significant cytotoxicity on iMSC3 and MG-63. The AgNPs were internalized in a concentration-dependent manner in both cell types. Additionally, the AgNPs exhibited a significant negative effect on osteogenesis but not on adipogenesis. C. conglomeratus-derived AgNPs had an impact on the differentiation capacity of iMSC3. Our results indicated that C. conglomeratus AgNPs and the associated phytochemicals could exhibit potential medical applications.

In situ metabolomic analysis of osteonecrosis of the femoral head (ONFH) using MALDI MSI.

Osteonecrosis of the femoral head (ONFH) is a common orthopedic disease characterized by disability and deformity. To better understand ONFH at molecular level and to explore the possibility of early diagnosis, instead of diagnosis based on macroscopic spatial characteristics, a matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) method was developed for ONFH disease for the first time. The most challenging step for ONFH MSI is to deal with human bone tissues which are much harder than the other biological samples studied by the reported MSI studies. In this work, the MSI sectioning method of hard bone tissues was established using tender acids and a series of test criteria. Small-molecule metabolites, such as lipids and amino acids, were detected in bone sections, realizing the in situ detection of spatial distribution of biometabolites.By comparing the distribution of metabolites from different regions of normal femoral head, ONFH bone tissue (ONBT), and adjacent ONFH bone tissue (ANBT), the whole process of femoral head from normal stage to necrosis was monitored and visualized at molecular level. Moreover, this developed MSI method was used for metabolomics study of ONFH. 72 differential metabolites were identified, suggesting that disturbances in energy metabolism and lipid metabolism affected the normal life activities of osteoblasts and osteoclasts. This study provides new perspectives for future pathological studies of ONFH.