Osteoblastic Bone Research Articles

Bone marrow endosteal stem cells dictate active osteogenesis and aggressive tumorigenesis

The bone marrow contains various populations of skeletal stem cells (SSCs) in the stromal compartment, which are important regulators of bone formation. It is well-described that leptin receptor (LepR)+ perivascular stromal cells provide a major source of bone-forming osteoblasts in adult and aged bone marrow. However, the identity of SSCs in young bone marrow and how they coordinate active bone formation remains unclear. Here we show that bone marrow endosteal SSCs are defined by fibroblast growth factor receptor 3 (Fgfr3) and osteoblast-chondrocyte transitional (OCT) identities with some characteristics of bone osteoblasts and chondrocytes. These Fgfr3-creER-marked endosteal stromal cells contribute to a stem cell fraction in young stages, which is later replaced by Lepr-cre-marked stromal cells in adult stages. Further, Fgfr3+ endosteal stromal cells give rise to aggressive osteosarcoma-like lesions upon loss of p53 tumor suppressor through unregulated self-renewal and aberrant osteogenic fates. Therefore, Fgfr3+ endosteal SSCs are abundant in young bone marrow and provide a robust source of osteoblasts, contributing to both normal and aberrant osteogenesis.

Hypoxia-induced Wnt/β-catenin signaling activation in subchondral bone osteoblasts leads to an osteoarthritis-like phenotype of chondrocytes in articular cartilage.

Background: Osteoarthritis (OA) is a whole-joint disease and characterized by alterations in the articular cartilage, subchondral bone, ligaments, and synovial membrane. The crosstalk between cartilage and subchondral bone plays a crucial role in the pathogenesis and progression of OA. Hypoxia has been reported to play an important role in cartilage degradation and subchondral bone remodeling in OA. In this study, we aimed to identify the involvement of hypoxia in modifying the osteoblast phenotypes and determine whether these alterations could influence the metabolism of chondrocytes. Methods: First, the levels of Hif-1α in subchondral bone of different compartments in patients with OA were assessed using immunohistochemistry (IHC). In in vitro, human primary osteoblasts were cultured under hypoxic and normoxic conditions, and the hypoxic or normoxic conditioned media (HCM and NCM) were used to culture human primary chondrocytes. Then, phenotypic changes in osteoblasts were assessed using reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Furthermore, the expression of type II collagen (COL2A1), aggrecan (ACAN), SRY-related high-mobility group-box gene 9 (SOX9), matrix metalloproteinase 13 (MMP13), and matrix metalloproteinase 3 (MMP3) in chondrocytes was measured using RT-PCR. Finally, the serum levels of Wnt-related proteins were determined using ELISA. Results: Hif-1α was significantly increased in severely sclerotic subchondral bone compared to less damaged subchondral bone. β-Catenin and SOST were identified as upregulated and downregulated in hypoxic osteoblasts, respectively. The hypoxia-induced results were confirmed by ELISA. Stimulating human primary chondrocytes with HCM significantly induced MMP13 and MMP3 and inhibited COL2A1, ACAN, and SOX9 mRNA expression. The serum levels of DKK-1 were significantly increased in human OA. Conclusion: Together, these findings revealed that hypoxia in subchondral bone is a key factor in the crosstalk between chondrocytes and osteoblasts and facilitates the shift of chondrocytes toward an OA-like phenotype probably by activating the Wnt/β-catenin signaling pathway in osteoblasts.

An Evaluation of Core Decompression and Cancellous Bone Grafting for Early-Stage Avascular Necrosis of the Femoral Head.

Background Avascular necrosis (AVN) is characterized by bone death due to impaired blood supply leading to its collapse causing pain and suboptimal joint function. The blood supply of the femoral head is so tenuous that even a slight vascular injury can predispose to AVN. Hence, AVN is commonly seen in the femoral head. Core decompression can arrest or even reverse the process of AVN and can avoid femoral head collapse and its sequelae. A lateral trochanteric approach is used for core decompression. The necrotic bone is removed from the femoral head. The use of a non-vascularized bone graft is more attractive than a vascularized graft because it is significantly less technically challenging. The regenerative properties due to the presence of osteoblasts in the trabecular bone and the ability to procure a large amount of graft make the iliac crest the gold standard site of cancellous bone graft harvesting. Core decompression can be considered an effective treatment modality in early-stage AVN(up to stage 2B) of the femoral head. Methodology A prospective, interventional study was conducted in a tertiary care teaching hospital in southern Rajasthan, India. 20 Patients with AVN of the femoral head (up to grade 2B of Ficat and Arlet classification) who met the inclusion and exclusion criteria and presented to the orthopedic outpatient department of our institute were included in this study. Patients were treated with core decompression and cancellous bone grafting with a graft taken from the iliac crest. The Harris Hip Score (HHS) and Visual Analog Scale (VAS) score were used to assess the outcomes. Results In our study, the majority (50%) of the patients were in the 20-30-year age group, making it the most common age group with a male predominance (85%). In this study, the final result was calculated according to the HHS and VAS scores. The mean HHS was 69.45 preoperatively and 83.55 at six months postoperatively. Similarly, the mean VAS score was 6.3 preoperatively and 3.8 at six months postoperatively. Conclusions Core decompression with cancellous bone grafting is a promising procedure in stages 1 and 2 as it reduces the symptoms in the majority of cases and improves functional outcomes.

Arl13b promotes the proliferation, migration, osteogenesis, and mechanosensation of osteoblasts

Primary cilia are microtubule-based organelles presenting on the surface of most postmitotic mammalian cells. As being signaling hubs and sensory organelles, primary cilia can respond to mechanical and chemical stimuli from the extracellular environment. Arl13b (ADP-ribosylation factor-like 13B), an atypical Arf/Arl family GTPase, was identified in genetic screening as a protein essential for maintaining the integrity of cilia and neural tubes. Previous studies on Arl13b have mostly focused on its role in the development of neural tubes, polycystic kidneys, and tumors, but no role in bone patterns was described. This study reported the essential roles of Arl13b in bone formation and osteogenic differentiation. Arl13b was highly expressed in bone tissues and osteoblasts, positively correlated with osteogenic activity during bone development. Furthermore, Arl13b was essential for primary cilium maintenance and Hedgehog signaling activation in osteoblasts. Arl13b knockdown in osteoblasts decreased the length of primary cilia and the upregulated levels of Gli1, Smo, and Ptch1 upon Smo agonist treatment. Additionally, Arl13b knockdown inhibited cell proliferation and migration. Moreover, Arl13b mediated osteogenesis and cell mechanosensation. Cyclic tension strain upregulated the Arl13b expression. Arl13b knockdown suppressed osteogenesis and mitigated cyclic tension strain-induced osteogenesis. These results suggest that Arl13b have important roles in bone formation and mechanosensation.

Relationship of the occurrence and main approaches to osteonecrosis of the jaw with the use of bisphosphonates: a concise systematic review

Introduction: In the setting of bucomaxillo facial surgery, bone tissue is a specialized connective tissue made up of cells and a mineralized extracellular matrix. To reduce the excessive resorption observed in these pathologies, it has been studied more rigorously in pre-clinical and clinical studies to improve anti-resorptive drugs that allow for treating or preventing pathologies of bone metabolism, such as bisphosphonates (BP) and denosumab (DN) as well as some angiogenesis inhibitors, can induce osteonecrosis of the mandible (ONJ). Objective:Foi realizar uma revisão sistemática concisa a respeito da relação da ocorrência e principais abordagens da osteonecrose da mandíbula e/ou maxilar com o uso dos bifosfonatos. Methods: The present study was followed by a systematic review model (PRISMA). The search strategy was performed in the PubMed, Cochrane Library, Web of Science and Scopus, and Google Scholar databases. The Cochrane Instrument was used to assess the risk of bias from the included studies. The present study was carried out from December 2022 to January of 2023. Results and Conclusion: A total of 108 articles were found. Initially, duplicate articles were excluded. After this process, the abstracts were evaluated and a new exclusion was performed based on the GRADE Instrument and Risk of Bias. A total of 55 articles were fully evaluated and 22 were included and discussed in this study. The American Association of Oral and Maxillofacial Surgeons proposed for the first time its nomenclature "Bisphosphonates Related Osteonecrosis of the Jaws". ONJ is the term used to describe bone cell death when the osteocyte becomes necrotic. Osteonecrosis of the jaws can be considered a severe adverse effect of BP therapy. Osteonecrosis of the jaws induced by the use of antiresorptive drugs commonly occurs in the oral cavity, mainly because the bone tissue is covered and protected only by a thin layer of periosteum and epithelium. Bisphosphonates inhibit RANKL expression, as well as stimulate OPG production by bone marrow cells and osteoblasts, inhibiting the RANK-RANKL interaction. These synergistic actions lead to a decrease in the recruitment of osteoclasts and, consequently, to a reduction in bone resorption.

Abstract 3609: Interaction of interleukin-6/8/18 with prostate cancer cell secreted growth factors are crucial for osteoblast proliferation and differentiation

Abstract There is currently no effective therapy for bone metastatic prostate cancer, which is the main cause of mortality in people with prostate cancer. Poor prognosis of bone metastasis has limitations in current therapeutics mainly due to the complexity of microenvironment. Our focus was on the interactions between prostate cancer cells and the bone microenvironment, including osteoclasts and osteoblasts in regard to growth factors and cytokines, as the bone microenvironment plays a significant role in this process. The present study shows the downstream effects of Stavudine (KB-S); a known antiviral compound along with 5-fluoro-1-((1R,4R)-4-((tetrahydro-2H-pyran-2-yl)oxy)cyclopent-2-en-1-yl)pyrimidine-2,4(1H,3H)-dione (KB-G) and 1-((1R,4R)-4-hydroxycyclopent-2-en-1-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (KB-H) on bone metastatic prostate cancer. Effects of KB-G/H and S on HEK-293 control cells were determined as moderate under tested conditions. A series of KB-G/H and S (10, 25, 50, 75 and 100 µM) was given for a 4-day period (with 24 h intervals) and the prostate cancer cell population was determined at the end of the 4-day treatment period. Preliminary results indicated that all 3 compounds significantly decreased DU-145 and PC-3 cell population compared to the effects on HEK-293 cells. Results suggested that all 3 compounds can decrease the prostate cancer cell progression by enhancing apoptosis. Our data also suggested that all 3 compounds downregulated interleukin (IL)-18, IL-8, IL-6 and C-X-C motif ligand-1(CXCL-1) in prostate cancer cells (PC-3 and DU-145). A human osteoblast cell line (hFOB 1.19) is being used to study the effect of cytokines in relation to transforming growth factor β (TGFβ), insulin-like growth factor (IGF) and platelet-derived growth factor (PDGF) that are secreted by prostate cancer cells. In-vitro migration/invasion assays, immunoprecipitation (IP), immunofluorescence microscope, cytokine profile array, real time qPCR and Western blot techniques are being used in this study to further analyze role of cytokines and growth factors on osteoblast differentiation. Our preliminary results suggest that all 3 compounds can be used to disrupt the main steps of prostate cancer bone metastasis. Results suggested that KB-G/H and S can be targeted to develop customized, tailored therapies for bone metastatic prostate cancer which merit further research. Citation Format: Aaron Jeshua Todman, Wishrawana S. Ratnayake, Luke Lajmi, Sloan Breedy, Kirpal Bisht, Mildred Acevedo-Duncan. Interaction of interleukin-6/8/18 with prostate cancer cell secreted growth factors are crucial for osteoblast proliferation and differentiation. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3609.

Label-free Quantitative Proteomic Analysis of Ascorbic Acid-induced Differentially Expressed Osteoblast-related Proteins in Dental Pulp Stem Cells from Deciduous and Permanent Teeth.

Proteomic is capable of elucidating complex biological systems through protein expression, function, and interaction under a particular condition. This study aimed to determine the potential of ascorbic acid alone in inducing differentially expressed osteoblast-related proteins in dental stem cells via the liquid chromatography-mass spectrometry/ mass spectrometry (LC-MS/MS) approach. The cells were isolated from deciduous (SHED) and permanent teeth (DPSC) and induced with 10 μg/mL of ascorbic acid. Bone mineralisation and osteoblast gene expression were determined using von Kossa staining and reverse transcriptase-polymerase chain reaction. The label-free protein samples were harvested on days 7 and 21, followed by protein identification and quantification using LC-MS/MS. Based on the similar protein expressed throughout treatment and controls for SHED and DPSC, overall biological processes followed by osteoblast-related protein abundance were determined using the PANTHER database. STRING database was performed to determine differentially expressed proteins as candidates for SHED and DPSC during osteoblast development. Both cells indicated brownish mineral stain and expression of osteoblast-related genes on day 21. Overall, a total of 700 proteins were similar among all treatments on days 7 and 21, with 482 proteins appearing in the PANTHER database. Osteoblast-related protein abundance indicated 31 and 14 proteins related to SHED and DPSC, respectively. Further analysis by the STRING database identified only 22 and 11 proteins from the respective group. Differential expressed analysis of similar proteins from these two groups revealed ACTN4 and ACTN1 as proteins involved in both SHED and DPSC. In addition, three (PSMD11/RPN11, PLS3, and CLIC1) and one (SYNCRIP) protein were differentially expressed specifically for SHED and DPSC, respectively. Proteome differential expression showed that ascorbic acid alone could induce osteoblastrelated proteins in SHED and DPSC and generate specific differentially expressed protein markers.

Magnetic Bone Tissue Engineering: Reviewing the Effects of Magnetic Stimulation on Bone Regeneration and Angiogenesis.

Bone tissue engineering emerged as a solution to treat critical bone defects, aiding in tissue regeneration and implant integration. Mainly, this field is based on the development of scaffolds and coatings that stimulate cells to proliferate and differentiate in order to create a biologically active bone substitute. In terms of materials, several polymeric and ceramic scaffolds have been developed and their properties tailored with the objective to promote bone regeneration. These scaffolds usually provide physical support for cells to adhere, while giving chemical and physical stimuli for cell proliferation and differentiation. Among the different cells that compose the bone tissue, osteoblasts, osteoclasts, stem cells, and endothelial cells are the most relevant in bone remodeling and regeneration, being the most studied in terms of scaffold-cell interactions. Besides the intrinsic properties of bone substitutes, magnetic stimulation has been recently described as an aid in bone regeneration. External magnetic stimulation induced additional physical stimulation in cells, which in combination with different scaffolds, can lead to a faster regeneration. This can be achieved by external magnetic fields alone, or by their combination with magnetic materials such as nanoparticles, biocomposites, and coatings. Thus, this review is designed to summarize the studies on magnetic stimulation for bone regeneration. While providing information regarding the effects of magnetic fields on cells involved in bone tissue, this review discusses the advances made regarding the combination of magnetic fields with magnetic nanoparticles, magnetic scaffolds, and coatings and their subsequent influence on cells to reach optimal bone regeneration. In conclusion, several research works suggest that magnetic fields may play a role in regulating the growth of blood vessels, which are critical for tissue healing and regeneration. While more research is needed to fully understand the relationship between magnetism, bone cells, and angiogenesis, these findings promise to develop new therapies and treatments for various conditions, from bone fractures to osteoporosis.

MMP14 cleaves PTH1R in the chondrocyte-derived osteoblast lineage, curbing signaling intensity for proper bone anabolism.

Bone homeostasis is regulated by hormones such as parathyroid hormone (PTH). While PTH can stimulate osteo-progenitor expansion and bone synthesis, how the PTH-signaling intensity in progenitors is controlled is unclear. Endochondral bone osteoblasts arise from perichondrium-derived osteoprogenitors and hypertrophic chondrocytes (HC). We found, via single-cell transcriptomics, that HC-descendent cells activate membrane-type 1 metalloproteinase 14 (MMP14) and the PTH pathway as they transition to osteoblasts in neonatal and adult mice. Unlike Mmp14 global knockouts, postnatal day 10 (p10) HC lineage-specific Mmp14 null mutants (Mmp14ΔHC) produce more bone. Mechanistically, MMP14 cleaves the extracellular domain of PTH1R, dampening PTH signaling, and consistent with the implied regulatory role, in Mmp14ΔHC mutants, PTH signaling is enhanced. We found that HC-derived osteoblasts contribute ~50% of osteogenesis promoted by treatment with PTH 1-34, and this response was amplified in Mmp14ΔHC. MMP14 control of PTH signaling likely applies also to both HC- and non-HC-derived osteoblasts because their transcriptomes are highly similar. Our study identifies a novel paradigm of MMP14 activity-mediated modulation of PTH signaling in the osteoblast lineage, contributing new insights into bone metabolism with therapeutic significance for bone-wasting diseases.

IOX-1 suppresses metastasis of osteosarcoma by upregulating histone H3 lysine trimethylation

New therapeutic approaches are needed for metastatic osteosarcoma (OS), as survival rates remain low despite surgery and chemotherapy. Epigenetic changes, such as histone H3 methylation, play key roles in many cancers including OS, although the underlying mechanisms are not clear. In this study, human OS tissue and OS cell lines displayed lower levels of histone H3 lysine trimethylation compared with normal bone tissue and osteoblast cells. Treating OS cells with the histone lysine demethylase inhibitor 5-carboxy-8-hydroxyquinoline (IOX-1) dose-dependently increased histone H3 methylation and inhibited cellular migratory and invasive capabilities, suppressed matrix metalloproteinase expression, reversed epithelial-to-mesenchymal transition by increasing levels of epithelial markers E-cadherin and ZO-1 and decreasing the expression of mesenchymal markers N-cadherin, vimentin, and TWIST, and also reduced stemness properties. An analysis of cultivated MG63 cisplatin-resistant (MG63-CR) cells revealed lower histone H3 lysine trimethylation levels compared with levels in MG63 cells. Exposing MG63-CR cells to IOX-1 increased histone H3 trimethylation and ATP-binding cassette transporter expression, potentially sensitizing MG63-CR cells to cisplatin. In conclusion, our study suggests that histone H3 lysine trimethylation is associated with metastatic OS and that IOX-1 or other epigenetic modulators present promising strategies to inhibit metastatic OS progression.

Molecular Basis beyond Interrelated Bone Resorption/Regeneration in Periodontal Diseases: A Concise Review.

Periodontitis is the sixth most common chronic inflammatory disease, destroying the tissues supporting the teeth. There are three distinct stages in periodontitis: infection, inflammation, and tissue destruction, where each stage has its own characteristics and hence its line of treatment. Illuminating the underlying mechanisms of alveolar bone loss is vital in the treatment of periodontitis to allow for subsequent reconstruction of the periodontium. Bone cells, including osteoclasts, osteoblasts, and bone marrow stromal cells, classically were thought to control bone destruction in periodontitis. Lately, osteocytes were found to assist in inflammation-related bone remodeling besides being able to initiate physiological bone remodeling. Furthermore, mesenchymal stem cells (MSCs) either transplanted or homed exhibit highly immunosuppressive properties, such as preventing monocytes/hematopoietic precursor differentiation and downregulating excessive release of inflammatory cytokines. In the early stages of bone regeneration, an acute inflammatory response is critical for the recruitment of MSCs, controlling their migration, and their differentiation. Later during bone remodeling, the interaction and balance between proinflammatory and anti-inflammatory cytokines could regulate MSC properties, resulting in either bone formation or bone resorption. This narrative review elaborates on the important interactions between inflammatory stimuli during periodontal diseases, bone cells, MSCs, and subsequent bone regeneration or bone resorption. Understanding these concepts will open up new possibilities for promoting bone regeneration and hindering bone loss caused by periodontal diseases.

Surface modifications of titanium dental implants with strontium eucommia ulmoides to enhance osseointegration and suppress inflammation

BackgroundTitanium (Ti) is now widely used as implant material due to its excellent mechanical properties and superior biocompatibilities, while its inert bioactivities might lead to insufficient osseointegration, and limit its performance in dental applications.MethodsWe introduced a robust and simple approach of modifying titanium surfaces with polysaccharide complexes. Titanium samples were subjected to hydrothermal treatment to create a uniform porous structure on the surface, followed by coating with a bioinspired and self-assembly polydopamine layer. Strontium Eucommia Ulmoides Polysaccharide (EUP-Sr) complexes are then introduced to the polydopamine-coated porous titanium. Multiple morphological and physiochemical characterizations are employed for material evaluation, while cell proliferation and gene expression tests using macrophages, primary alveolar bone osteoblasts, and vascular endothelial cells are used to provide an overall insight into the functions of the product. The significances of statistical differences were analyzed using student’s t-test.ResultsMicroscopic and spectrometric characterizations confirmed that the Ti surface formed a porous structure with an adequate amount of EUP-Sr loading. The attachment was attributed to hydrogen bonding between the ubiquitous glycosidic linkage of the polysaccharide complex and the ring structure of polydopamine, yet the loaded EUP-Sr complex can be gradually released, consequently benefiting the neighboring microenvironment. Cell experiments showed no cytotoxicity of the material, and the product showed promising anti-inflammation, osseointegration, and angiogenesis properties, which were further confirmed by in vivo evaluations.ConclusionWe believe the EUP-Sr modified titanium implant is a promising candidate to be used in dental applications with notable osteoimmunomodulation and angiogenesis functions. And the novel technique proposed in this study would benefit the modification of metal/inorganic surfaces with polysaccharides for future research.

Expression Changes of Hypoxia-Inducible Factor-1α in G-CSF Induced Hematopoietic Stem Cell Mobilization

To investigate the expression and its relative mechanism of hypoxia-inducible factor-1α(HIF-1α) in bone marrow(BM) of mice during G-CSF mobilization of hematopoietic stem cells (HSC) . Flow cytometry was used to detect the proportion of Lin-Sca-1+ c-kit+ (LSK) cells in peripheral blood of C57BL/6J mice before and after G-CSF mobilization. And the expression of HIF-1α and osteocalcin (OCN) mRNA and protein were detected by RQ-PCR and immunohistochemistry. The number of osteoblasts in bone marrow specimens of mice was counted under the microscope. The proportion of LSK cells in peripheral blood began to increase at day 4 of G-CSF mobilization, and reached the peak at day 5, which was significantly higher than that of control group (P<0.05). There was no distinct difference in the expression of HIF-1α mRNA between bone marrow nucleated cells and osteoblasts of steady-state mice (P=0.073), while OCN mRNA was mainly expressed in osteoblasts, which was higher than that in bone marrow nucleated cells (P=0.034). After mobilization, the expression level of HIF-1α increased, but OCN decreased, and the number of endosteum osteoblasts decreased. The change of HIF-1α expression was later than that of OCN and was consistent with the proportion of LSK cells in peripheral blood. The expression of HIF-1α in bone marrow was increased during the mobilization of HSC mediated by G-CSF, and one of the mechanisms may be related to the peripheral migration of HSC induced by osteoblasts inhibition.

Effect of Hydrogen Peroxide on the Surface and Attractiveness of Various Zirconia Implant Materials on Human Osteoblasts: An In Vitro Study.

The aim of this in vitro study was to investigate the effect of hydrogen peroxide (H2O2) on the surface properties of various zirconia-based dental implant materials and the response of human alveolar bone osteoblasts. For this purpose, discs of two zirconia-based materials with smooth and roughened surfaces were immersed in 20% H2O2 for two hours. Scanning electron and atomic force microscopy showed no topographic changes after H2O2-treatment. Contact angle measurements (1), X-ray photoelectron spectroscopy (2) and X-ray diffraction (3) indicated that H2O2-treated surfaces (1) increased in hydrophilicity (p < 0.05) and (2) on three surfaces the carbon content decreased (33-60%), while (3) the monoclinic phase increased on all surfaces. Immunofluorescence analysis of the cell area and DNA-quantification and alkaline phosphatase activity revealed no effect of H2O2-treatment on cell behavior. Proliferation activity was significantly higher on three of the four untreated surfaces, especially on the smooth surfaces (p < 0.05). Within the limitations of this study, it can be concluded that exposure of zirconia surfaces to 20% H2O2 for 2 h increases the wettability of the surfaces, but also seems to increase the monoclinic phase, especially on roughened surfaces, which can be considered detrimental to material stability. Moreover, the H2O2-treatment has no influence on osteoblast behavior.

Epigenetic Repression of RUNX2 and OSX Promoters Controls the Nonmineralized State of the Periodontal Ligament.

The nonmineralized state of the mammalian periodontal ligament is one of the hallmarks of vertebrate evolution as it provides resilient and nontraumatic tooth anchorage for effective predation. Here we sought to determine how the chromatin state of key mineralization gene promoters contributes to the nonmineralized periodontal ligament in the midst of fully mineralized alveolar bone and cementum anchor tissues. In developing mouse periodontal tissues, RUNX2 was localized to alveolar bone-lining cells, while OSX was localized throughout the periodontal ligament's soft tissue. Matching RT-PCR amplification data and western blot comparisons demonstrated that the expression of RUNX2 and OSX bone mineralization transcription factors was at least 2.5-fold elevated in alveolar bone osteoblasts versus periodontal ligament fibroblasts. ChIP enrichment data along the RUNX2 and OSX promoters revealed increased H3K4me3 marks in alveolar bone osteoblasts, while H3K9me3 and H3K27me3 marks were elevated in periodontal ligament fibroblasts. In support of an epigenetic mechanism responsible for the inhibition of mineralization gene expression in periodontal progenitors, histone methylation inhibitors DZNep and Chaetocin reactivated RUNX2 and OSX expression in periodontal progenitors and increased alkaline phosphatase and Alizarin Red, while the in vivo application of DZNep in rat maxillae resulted in aberrant mineralization in the periodontal ligament and a narrowing of the nonmineralized periodontal space. Together, these studies demonstrate that the nonmineralized state of the mammalian periodontal ligament is controlled by an epigenetic regulation of the RUNX2 and OSX key mineralization gene promoters.

Changes in Hox Gene Chromatin Organization during Odontogenic Lineage Specification.

Craniofacial tissues comprise highly evolved organs characterized by a relative lack of expression in the HOX family transcription factors. In the present study, we sought to define the epigenetic events that limit HOX gene expression from undifferentiated neural crest cells to semi-differentiated odontogenic progenitors and to explore the effects of elevated levels of HOX. The ChIP-chip data demonstrated high levels of repressive H3K27me3 marks on the HOX gene promoters in ES and cranial neural crest cells when compared to the H3K4me3 marks, while the K4/K27 ratio was less repressive in the odontogenic progenitors, dental follicle, dental pulp, periodontal ligament fibroblasts, alveolar bone osteoblasts, and cementoblasts. The gene expression of multiple HOX genes, especially those from the HOXA and HOXB clusters, was significantly elevated and many times higher in alveolar bone cells than in the dental follicle cells. In addition, the HOX levels in the skeletal osteoblasts were many times higher in the trunk osteoblasts compared to the alveolar bone osteoblasts, and the repressive mark H3K27me3 promoter occupancy was substantially and significantly elevated in the alveolar bone osteoblasts when compared to the trunk osteoblasts. To explore the effect of elevated HOX levels in craniofacial neural crest cells, HOX expression was induced by transfecting cells with the Cdx4 transcription factor, resulting in a significant decrease in the mineralization markers, RUNX2, OSX, and OCN upon HOX elevation. Promoting HOX gene expression in developing teeth using the small molecule EZH2 inhibitor GSK126 resulted in an increased number of patterning events, supernumerary cusp formation, and increased Hoxa4 and Hoxb6 gene expression when compared to the controls. Together, these studies illustrate the profound effects of epigenetic regulatory events at all stages of the differentiation of craniofacial peripheral tissues from the neural crest, including lineage specification, tissue differentiation, and patterning.

Calcium-Deficit Diet Improves Iron Content in Ovariectomized Rats

In women, menopause is associated with disorders related to calcium and iron content, which may increase the risk of osteoporosis. This study aimed to determine the effect of calcium deficiency on the iron content in ovariectomized rats. This study included 30 3-month-old female rats, which were divided into three groups: group C (n = 10)—control group fed the standard diet; group O—ovariectomized rats fed the standard diet; and group D—ovariectomized rats fed the calcium-deficit diet. After 3 months of experimental intervention, the weight of the rats was measured, and blood and tissue samples were collected. Morphological parameters were analyzed in whole blood, and serum levels of leptin, estrogen and C-reactive protein, and total antioxidant status were determined. The iron content was measured in tissues, and histological analysis was performed in the femur. The results obtained demonstrated that ovariectomy significantly decreased the iron content in bones, hair, spleen, liver, and kidneys. The calcium-deficit diet increased the iron content in tissues and the hemoglobin level in ovariectomized rats and also enhanced the number of osteoblasts in bones compared with the O group. In conclusion, calcium deficiency improved the iron content in ovariectomized rats in this 12-week study.

The ADAM9/WISP-1 axis cooperates with osteoblasts to stimulate primary prostate tumor growth and metastasis.

Background: Metastatic prostate cancer (PCa) predicts a poor prognosis and lower likelihood of survival. Osteoblasts (OBs) are known to be responsible for the synthesis and mineralization of bone, although it is unclear as to whether PCa in the prostate gland cooperates with OBs in bone to promote PCa malignant transformation. We aimed to elucidate how primary PCa cells cooperate with distal OBs and contribute to the vicious cycle that leads to metastatic PCa. Methods: N-cadherin, E-cadherin, and Twist protein expression were measured by Western blot. Twist translocation into the nucleus was detected by the immunofluorescence (IF) assay. Enzyme-linked immunosorbent assay (ELISA) detected protein levels in human serum samples. Levels of candidate protein expression were examined by the human cytokine array. Prostate tumor growth and metastasis were analyzed by orthotopic and metastatic prostate cancer models, respectively. Immunohistochemistry (IHC) staining was used to observe ADAM metallopeptidase domain 9 (ADAM9) and WNT1 inducible signaling pathway protein 1 (WISP-1) expression in tissue. Results: Our in vitro and in vivo analyses have now discovered that primary PCa expressing ADAM9 protein enables the transformation of OBs into PCa-associated osteoblasts (PCa-OBs), inducing WISP-1 secretion from PCa-OBs in the bone microenvironment. The upregulation of WISP-1 in bone provided feedback to primary PCa and promoted PCa cell aggressiveness via epithelial-mesenchymal transition (EMT) activity. Elevated levels of WISP-1 expression were detected in the serum of patients with PCa. ADAM9 levels were overexpressed in tumor tissue from PCa patients; ADAM9 blockade interrupted OB-induced release of WISP-1 and also suppressed primary tumor growth and distal metastasis in orthotopic PCa mouse models. Conclusion: Our study suggests that the ADAM9/WISP-1 axis assists with metastatic PCa progression. Thus, targeting the ADAM9/WISP-1 axis may help to prevent the malignant phenotypes of PCa cells.

Қазақтандағы жетілмеген остеогенезі бар балаларды хирургиялық емдеуде отандық интрамедуллярлық телескопиялық жүйенің күтілетін үлесі

Osteogenesis imperfecta (OI) is a group of inherited connective tissue disorders in collagen structure, processing or cross-linking, new bone formation and osteoblast differentiation, which result in fragility of the bones, recurrent fractures and deformities.The article presents scientific data on the types and features of metal structures used in intramedullary osteotomy performed for the surgical treatment of fractures and deformities of long tubular bones with osteogenesis imperfecta in children and outlines the benefits of the developed domestic intramedullary telescopic system "DOSSANOV" in providing surgical treatment to children with osteogenesis imperfecta. For the "crystal children", whose number is growing in Kazakhstan, the expected advantages of using this system are analyzed.Key words: Osteogenesis imperfecta, children, intramedullary osteosynthesis, osteotomy, intramedullary rods, intramedullary telescopic system.

Fractional Order Nonlinear Bone Remodeling Dynamics Using the Supervised Neural Network

This study aims to solve the nonlinear fractional-order mathematical model (FOMM) by using the normal and dysregulated bone remodeling of the myeloma bone disease (MBD). For the more precise performance of the model, fractional-order derivatives have been used to solve the disease model numerically. The FOMM is preliminarily designed to focus on the critical interactions between bone resorption or osteoclasts (OC) and bone formation or osteoblasts (OB). The connections of OC and OB are represented by a nonlinear differential system based on the cellular components, which depict stable fluctuation in the usual bone case and unstable fluctuation through the MBD. Untreated myeloma causes by increasing the OC and reducing the osteoblasts, resulting in net bone waste the tumor growth. The solutions of the FOMM will be provided by using the stochastic framework based on the Levenberg-Marquardt backpropagation (LVMBP) neural networks (NN), i.e., LVMBPNN. The mathematical performances of three variations of the fractional-order derivative based on the nonlinear disease model using the LVMPNN. The static structural performances are 82% for investigation and 9% for both learning and certification. The performances of the LVMBPNN are authenticated by using the results of the Adams-Bashforth-Moulton mechanism. To accomplish the capability, steadiness, accuracy, and ability of the LVMBPNN, the performances of the error histograms (EHs), mean square error (MSE), recurrence, and state transitions (STs) will be provided.