The unbearable lightness of bone marrow homeostasis

The anatomical and functional dimensions of bone marrow topography have been at the forefront of modern bone and immunological research for many years and remain a source of complexity and perplexity due to the multitude of microhabitats within this microenvironment. In fact, research has uncovered fascinating functional aspects of bone marrow residents, and the bone marrow niche has been identified as the foremost reservoir of a variety of cells including hematopoietic, skeletal and endothelial stem/progenitor cells. The physical interactions of the marrow residents, combined with the release of cytokines and growth factors, organize well-defined operative compartments, which preserve bone and immune cell homeostasis. In a simplistic view, both the hematopoietic and bone marrow stromal (mesenchymal) stem/progenitor cell populations dwell at the interface between the endosteum and the bone marrow area (endosteal niche) and in the perivascular space (vascular niche). Indeed, the tantalizing hypothesis of bone marrow regulatory dependency on these niches is supported by current research insofar as the increase in the number of osteoblasts results in a concomitant increase in the hematopoietic population, indicating that the osteoblasts and the endosteal niche are key components of HSC maintenance. On the other hand, impaired function of the vascular niche compromises the endosteal niche's ability to support hematopoiesis. These fascinating discoveries indicate that there are strong ties between bone marrow inhabitants within the confines of the bone marrow itself. When these ties fail, niche-niche communication suffers and results in reduced bone formation, enfeebled hematopoiesis and unrestrained HSC migration through blood circulation. This study focused on the extraordinary homeostatic equilibrium and function of both bone and immune cells within the spatially defined microenvironment of bone marrow. But how important is the anatomically outlined scenery in which the bone marrow entity supports and hosts the hematopoietic elements

Photobiomodulation by Near-Infrared 980-nm Wavelengths Regulates Pre-Osteoblast Proliferation and Viability through the PI3K/Akt/Bcl-2 Pathway

Background: bone tissue regeneration remains a current challenge. A growing body of evidence shows that mitochondrial dysfunction impairs osteogenesis and that this organelle may be the target for new therapeutic options. Current literature illustrates that red and near-infrared light can affect the key cellular pathways of all life forms through interactions with photoacceptors within the cells' mitochondria. The current study aims to provide an understanding of the mechanisms by which photobiomodulation (PBM) by 900-nm wavelengths can induce in vitro molecular changes in pre-osteoblasts. Methods: The PubMed, Scopus, Cochrane, and Scholar databases were used. The manuscripts included in the narrative review were selected according to inclusion and exclusion criteria. The new experimental set-up was based on irradiation with a 980-nm laser and a hand-piece with a standard Gaussian and flat-top beam profile. MC3T3-E1 pre-osteoblasts were irradiated at 0.75, 0.45, and 0.20 W in continuous-wave emission mode for 60 s (spot-size 1 cm2) and allowed to generate a power density of 0.75, 0.45, and 0.20 W/cm2 and a fluence of 45, 27, and 12 J/cm2, respectively. The frequency of irradiation was once, three times (alternate days), or five times (every day) per week for two consecutive weeks. Differentiation, proliferation, and cell viability and their markers were investigated by immunoblotting, immunolabelling, fluorescein-FragELTM-DNA, Hoechst staining, and metabolic activity assays. Results and conclusions: The 980-nm wavelength can photobiomodulate the pre-osteoblasts, regulating their metabolic schedule. The cellular signal activated by 45 J/cm2, 0.75 W and 0.75 W/cm2 consist of the PI3K/Akt/Bcl-2 pathway; differentiation markers were not affected, nor do other parameters seem to stimulate the cells. Our previous and present data consistently support the window effect of 980 nm, which has also been described in extracted mitochondria, through activation of signalling PI3K/Akt/Bcl-2 and cyclin family, while the Wnt and Smads 2/3-β-catenin pathway was induced by 55 J/cm2, 0.9 W and 0.9 W/cm

Administration of DNA Plasmid Coding Protein Aggregating Domain Induces Inflammatory Bone Loss.

Background: Plasmids coding protein aggregation polypeptides from different sources have beenproposed as genetic adjuvants for DNA vaccines. We reported that a plasmid (pATRex), encompassing the DNA sequence for the von Willebrand A (vWA/A) domain of the Anthrax Toxin Receptor-1 (ANTXR-1, alias TEM8, Tumor Endothelial Marker 8), acts as strong immune adjuvant by inducing formation of insoluble intracellular aggregates and subsequent cell death. Aims: In the present study we addressed the question of whether there is any substantial immunotoxicity associated with the use of self-aggregating proteins as genetic adjuvants. Results: Here we report, by mean of histology, X-ray and molecular examinations of bone specimens, the unexpected finding that intramuscular injection of pATRex in mice triggers, per se, severe bone loss (osteoporosis) independently from the sex and genotype of the treated animals. Conclusion: Even though the study suggests that proteinaceous “sticky “ adjuvants are unlikely to find their way into practical vaccination, the information gained is of value as ATRex injections could provide an additional, simplified, mouse model of osteoporosis. Moreover, our results provide an experimental support to the hypothesis that proteotoxic aggregates chronically activate the innate immune system in amyloid and aggregosome disorders

Tackling Inequalities in Oral Health: Bone Augmentation in Dental Surgery through the 3D Printing of Poly(ε-caprolactone) Combined with 20% Tricalcium Phosphate

Personalized medicine and overcoming healthcare inequalities have gained significant popularity in recent years. Polymers offer an ideal solution due to their cost-effectiveness, ease of customized 3D printing, and potential for wide-scale expansion. Poly- mers blended with β-tricalcium phosphate (TCP) have been found to synergize with the environ- mental tissues of maxillary bones and promote osteoconductivity. However, little is known about their properties after printing and their ability to maintain their biological role; additionally, limi- tations exist in 3D printing when high TPC concentrations are added. Our study demonstrated that poly ε-caprolactone (PCL)+β-TCP 20% composite can be successfully printed and is a suitable ma- terial for commercial 3D printing. The material also demonstrated biocompatibility, supporting osteoblast adhesion and promoting cell proliferation and differentiation. The composite can also sustain ISO14937:200935 sterilization procedures, which makes it an ideal material for printing medical devices that can be used by clinicians worldwide

INF-γ encoding plasmid administration triggers bone loss and disrupts bone marrow microenvironment

IFN-γ is a pleotropic cytokine produced in the bone microenvironment. Although IFN-γ is known to play a critical role on bone remodeling, its function is not fully elucidated. Consistently, outcomes on the effects of IFN-γ recombinant protein on bone loss are contradictory among reports. In our work we explored, for the first time, the role of IFN-γ encoding plasmid (pIFN-γ) in a mouse model of osteopenia induced by ovariectomy and in the sham-operated counterpart to estimate its effects in skeletal homeostasis. Ovariectomy produced a dramatic decrease of bone mineral density (BMD). pINF-γ injected mice showed a pathologic bone and bone marrow phenotype; the disrupted cortical and trabecular bone microarchitecture was accompanied by an increased release of pro-inflammatory cytokine by bone marrow cells. Moreover, mesenchymal stem cells’ (MSCs) commitment to osteoblast was found impaired, as evidenced by the decline of osterix-positive (Osx+) cells within the mid-diaphyseal area of femurs. For instance, a reduction and redistribution of CXCL12 cells have been found, in accordance with bone marrow morphological alterations. As similar effects were observed both in sham-operated and in ovariectomized mice, our studies proved that an increased IFN-γ synthesis in bone marrow might be sufficient to induce inflammatory and catabolic responses even in the absence of pathologic predisposing substrates. In addition, the obtained data might raise questions about pIFN-γ’s safety when it is used as vaccine adjuvant

A Comparative Study Between the Effectiveness of 980 nm Photobiomodulation Delivered by Hand-Piece With Gaussian vs. Flat-Top Profiles on Osteoblasts Maturation

Photobiomodulation (PBM) is a clinically accepted tool in regenerative medicine and dentistry to improve tissue healing and repair and to restore the functional disability. The current in vitro study aimed to investigate the photobiomodulatory effects of 980 nm wavelength (the real energy at the target: ~0.9 W, ~0.9 W/cm2, 60 s, ~55 J/cm2 and a single energy ~55 J in CW) on MC3T3-E1 pre-osteoblast, delivered with flattop profile in comparison to the standard profile. The laser groupings and their associated energies were: Group 1 - once per week (total energy 110 J); Group 2 - three times per week (alternate day) (total energy 330 J); Group 3 - five times per week (total energy 550 J). The metabolic activity and the osteoblasts maturation were analyzed by alkaline phosphatase assay, alizarin red S histological staining, immunoblot and/or double immunolabeling analysis for Bcl2, Bax, Runx-2, Osx, Dlx5, osteocalcin, and collagen Type 1. Our data, for the first time, prove that laser irradiation of 980 nm wavelength with flat-top beam profile delivery system, compared to standard-Gaussian profile, has improved photobiomodulatory efficacy on pre-osteoblastic cells differentiation. Mechanistically, the irradiation enhances the pre-osteoblast differentiation through activation of Wnt signaling and activation of Smads 2/3-βcatenin pathway