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Background: Osteosarcoma (OS) survival rates and outcome have not improved in 50 years since the advent of modern chemotherapeutics. Thus, there is a critical need for an improved understanding of the tumor microenvironment to identify better therapies. Extracellular matrix (ECM) deposition and hypoxia are known to abrogate the efficacy of various chemical and cell‐based therapeutics. Here, we aim to mechanistically investigate the combinatorial effects of hypoxia and matrix deposition with the use of OS spheroids. Methods: We use two murine OS cell lines with differential metastatic potential to form spheroids. We form spheroids of two sizes, use ascorbate‐2‐phosphate supplementation to enhance ECM deposition, and study cell response under standard (21% O2) and physiologic (5% O2) oxygen tensions. Finally, we examine chemotherapeutic responses to doxorubicin treatment. Results: ECM production and oxygen tension are key determinants of spheroid size through cell organization based on nutrient and oxygen distribution. Interestingly, highly metastatic OS is more susceptible to chemotherapeutics compared to less metastatic OS when matrix production increases. Together, these data suggest that dynamic interactions between ECM production and oxygen diffusion may result in distinct chemotherapeutic responses despite inherent tumor aggressiveness. Conclusion: This work establishes OS spheroids as a valuable tool for early OS tumor formation investigation and holds potential for novel therapeutic target and prognostic indicator discovery. [ABSTRACT FROM AUTHOR]

The biophysical heterogeneity of the bone–cartilage interface requires complex materials to mimic differences in bone density, extracellular matrix composition, and mineralization. Biomaterial approaches to repair osteochondral tissue typically use multilayer scaffolds, which require multistep fabrication and may undergo delamination at the construct interface. This work describes the development of functionalized microgels for the repair of osteochondral tissues using an N‐cadherin peptide, bone morphogenetic protein‐2 (BMP‐2) peptide, and changes in stiffness to create pro‐osteogenic and prochondrogenic microgels. Microgels, when annealed into a scaffold, outperforms bulk hydrogel controls evidenced by upregulation of osteogenic and chondrogenic markers in mesenchymal stromal cells (MSCs). The macroporous void space present in microgel anneals scaffolds enabled robust cell proliferation and extracellular matrix (ECM) deposition throughout the entire scaffold. A bilayer functionalized annealed microgel scaffold is then created and the ability to spatially control the differentiation of MSCs is assessed. Osteochondral bilayer scaffolds exhibit distinct regions of osteogenic and chondrogenic protein expression as a function of microgel population upon immunostaining for osteocalcin and aggrecan, respectively. Spatial transcriptomics confirm osteogenic and chondrogenic genes are upregulated in their respective microgel regions. These studies highlight the tunable and functionalizable nature of microgels and the importance of macroporous void space. [ABSTRACT FROM AUTHOR]

The energetic positions of the two Fe dopant levels in the bandgap of β-Ga2O3 are determined to be at 3.05(±0.05) and 3.85(±0.05) eV below the conduction band minimum from transmission measurements employing a sub-picosecond tunable ultraviolet laser radiation source. A further measurement of the quantum efficiency of photoelectron emission from the Fe:Ga2O3(010) photocathode is consistent with the obtained absorption data and a dominant optical phonon assisted Franck–Condon emission mechanism, while also providing an estimate of 100 ps for the conduction band electron lifetime. [ABSTRACT FROM AUTHOR]

Metastasis is the principal factor in poor prognosis for individuals with osteosarcoma (OS). Understanding the events that lead to metastasis is critical to develop better interventions for this disease. Alveolar macrophages are potentially involved in priming the lung microenvironment for OS metastasis, yet the mechanisms involved in this process remain unclear. Since extracellular vesicles (EVs) are a known actor in primary tumor development, their potential role in OS metastagenesis through macrophage modulation is explored here. The interaction of EVs isolated from highly metastatic (K7M2) and less metastatic (K12) osteosarcoma cell lines is compared with a peritoneal macrophage cell line. An EV concentration that reproducibly induced macrophage migration is identified first, then used for later experiments. By confocal microscopy, both EV types associated with M0 or M1 macrophages; however, only K7M2‐EVs are associated with M2 macrophages, an interaction that is abrogated by EV pre‐treatment with anti‐CD47 antibody. Interestingly, all interactions appeared to be surface binding, not internalized. In functional studies, K7M2‐EVs polarized fewer macrophages to M1. Together, these data suggest that K7M2‐EVs have unique interactions with macrophages that can contribute to the production of a higher proportion of pro‐tumor type macrophages, thereby accelerating metastasis. [ABSTRACT FROM AUTHOR]

Mesenchymal stromal cells (MSCs) are a promising cell population for musculoskeletal cell‐based therapies due to their multipotent differentiation capacity and complex secretome. Cells from younger donors are mechanosensitive, evidenced by changes in cell morphology, adhesivity, and differentiation as a function of substrate stiffness in both two‐ and three‐dimensional culture. However, MSCs from older individuals exhibit reduced differentiation potential and increased senescence, limiting their potential for autologous use. While substrate stiffness is known to modulate cell phenotype, the influence of the mechanical environment on senescent MSCs is poorly described. To address this question, we cultured irradiation induced premature senescent MSCs on polyacrylamide hydrogels and assessed expression of senescent markers, cell morphology, and secretion of inflammatory cytokines. Compared to cells on tissue culture plastic, senescent MSCs exhibited decreased markers of the senescence associated secretory phenotype (SASP) when cultured on 50 kPa gels, yet common markers of senescence (e.g., p21, CDKN2A, CDKN1A) were unaffected. These effects were muted in a physiologically relevant heterotypic mix of healthy and senescent MSCs. Conditioned media from senescent MSCs on compliant substrates increased osteoblast mineralization compared to conditioned media from cells on TCP. Mixed populations of senescent and healthy cells induced similar levels of osteoblast mineralization compared to healthy MSCs, further indicating an attenuation of the senescent phenotype in heterotypic populations. These data indicate that senescent MSCs exhibit a decrease in senescent phenotype when cultured on compliant substrates, which may be leveraged to improve autologous cell therapies for older donors. [ABSTRACT FROM AUTHOR]

Mesenchymal stromal cells (MSCs) are widely used in cell-based therapies and tissue regeneration for their potent secretome, which promotes host cell recruitment and modulates inflammation. Compared to monodisperse cells, MSC spheroids exhibit improved viability and increased secretion of immunomodulatory cytokines. While mechanical stimulation of monodisperse cells can increase cytokine production, the influence of mechanical loading on MSC spheroids is unknown. Here, we evaluated the effect of controlled, uniaxial cyclic compression on the secretion of immunomodulatory cytokines by human MSC spheroids and tested the influence of load-induced gene expression on MSC mechanoresponsiveness. We exposed MSC spheroids, entrapped in alginate hydrogels, to three cyclic compressive regimes with varying stress (L) magnitudes (i.e., 5 and 10 kPa) and hold (H) durations (i.e., 30 and 250 s) L5H30, L10H30, and L10H250. We observed changes in cytokine and chemokine expression dependent on the loading regime, where higher stress regimes tended to result in more exaggerated changes. However, only MSC spheroids exposed to L10H30 induced human THP-1 macrophage polarization toward an M2 phenotype compared to static conditions. Static and L10H30 loading facilitated a strong, interlinked F-actin arrangement, while L5H30 and L10H250 disrupted the structure of actin filaments. This was further examined when the actin cytoskeleton was disrupted via Y-27632. We observed downregulation of YAP-related genes, and the levels of secreted inflammatory cytokines were globally decreased. These findings emphasize the essential role of mechanosignaling in mediating the immunomodulatory potential of MSC spheroids. [ABSTRACT FROM AUTHOR]

Concentration – discharge (C – Q) relationships in streamflow can serve a variety of purposes, including elucidating catchment water sources, quantifying chemical weathering, and estimating solute fluxes. Four main C – Q relationships have been identified: dilution (decreasing C with increasing Q), flushing (increasing C with increasing Q), chemostatic (little or no change in C with increasing Q) and chemostochastic (great variation in C relative to that of Q) behaviour. We used a 40‐year data record for 10 headwater forested catchments in central Ontario, Canada, to examine C – Q relationships for a range of geogenic, exogenic and bioactive solutes. The thin overburden cover of this landscape and general absence of deep groundwater contributions to streamflow suggest that streamflow solute concentrations and their relationship with discharge would be sensitive to pronounced changes in climate and atmospheric deposition chemistry over the monitoring period. We explored the temporal and spatial stability of C – Q relationships in response to these changes at annual and seasonal timescales for relatively wet and dry conditions and for periods with relatively high and low atmospheric deposition. Chemostasis dominated C – Q relationships at the annual and seasonal scales for geogenic (e.g., SiO2) and exogenic (e.g., Cl−) solutes, although bioactive solutes (e.g., NO3−, NH4+) exhibited non‐chemostatic behaviour for some catchments, time periods and wetness conditions. C – Q relationships derived at the annual timescale often masked pronounced seasonal shifts in bioactive solute concentration response to changes in streamflow and provide only partial insight into the production and delivery of such solutes in this forest landscape. Relatively subtle changes in overburden thickness across this landscape appeared to explain some of the deviations from widespread chemostasis. Nevertheless, this general chemostatic behaviour was largely temporally stable across catchments and solutes, highlighting its resilience in the face of such non‐stationary influences as climate conditions and atmospheric deposition. [ABSTRACT FROM AUTHOR]

Prostate cancer (PCa) is an increasingly prevalent health problem in the developed world. Effective treatment options exist for localized PCa, but metastatic PCa has fewer treatment options and shorter patient survival. PCa and bone health are strongly entwined, as PCa commonly metastasizes to the skeleton. Since androgen receptor signaling drives PCa growth, androgen-deprivation therapy whose sequelae reduce bone strength constitutes the foundation of advanced PCa treatment. The homeostatic process of bone remodeling -- produced by concerted actions of bone-building osteoblasts, bone-resorbing osteoclasts, and regulatory osteocytes -- may also be subverted by PCa to promote metastatic growth. Mechanisms driving skeletal development and homeostasis, such as regional hypoxia or matrix-embedded growth factors, may be subjugated by bone metastatic PCa. In this way, the biology that sustains bone is integrated into adaptive mechanisms for the growth and survival of PCa in bone. Skeletally metastatic PCa is difficult to investigate due to the entwined nature of bone biology and cancer biology. Herein, we survey PCa from origin, presentation, and clinical treatment to bone composition and structure and molecular mediators of PCa metastasis to bone. Our intent is to quickly yet effectively reduce barriers to team science across multiple disciplines that focuses on PCa and metastatic bone disease. We also introduce concepts of tissue engineering as a novel perspective to model, capture, and study complex cancer-microenvironment interactions. [ABSTRACT FROM AUTHOR]

The field of biomaterials science is highly active, with a steadily increasing number of publications and new journals being founded. This article brings together contributions from the editors of six leading journals in the area of biomaterials science and engineering. Each contributor highlights specific advances, topics, and trends that have emerged through the publications in their respective journal in the calendar year 2022. It presents a global perspective on a wide range of material types, functionalities, and applications. The highlighted topics include a diversity of biomaterials; from proteins, polysaccharides, and lipids to ceramics, metals, advanced composites, and a variety of new forms of these materials. Important advances in dynamically functional materials are presented, including a range of fabrication techniques such as bioassembly, 3D bioprinting and microgel formation. Similarly, several applications are highlighted in drug and gene delivery, biological sensing, cell guidance, immunoengineering, electroconductivity, wound healing, infection resistance, tissue engineering, and treatment of cancer. The goal of this paper is to provide the reader with both a broad view of recent biomaterials research, as well as expert commentary on some of the key advances that will shape the future of biomaterials science and engineering. [ABSTRACT FROM AUTHOR]

Cell therapies offer a tailorable, personalized treatment for use in tissue engineering to address defects arising from trauma, inefficient wound repair, or congenital malformation. However, most cell therapies have achieved limited success to date. Typically injected in solution as monodispersed cells, transplanted cells exhibit rapid cell death or insufficient retention at the site, thereby limiting their intended effects to only a few days. Spheroids, which are dense, three-dimensional (3D) aggregates of cells, enhance the beneficial effects of cell therapies by increasing and prolonging cell–cell and cell–matrix signaling. The use of spheroids is currently under investigation for many cell types. Among cells under evaluation, spheroids formed of mesenchymal stromal cells (MSCs) are particularly promising. MSC spheroids not only exhibit increased cell survival and retained differentiation, but they also secrete a potent secretome that promotes angiogenesis, reduces inflammation, and attracts endogenous host cells to promote tissue regeneration and repair. However, the clinical translation of spheroids has lagged behind promising preclinical outcomes due to hurdles in their formation, instruction, and use that have yet to be overcome. This review will describe the current state of preclinical spheroid research and highlight two key examples of spheroid use in clinically relevant disease modeling. It will highlight techniques used to instruct the phenotype and function of spheroids, describe current limitations to their use, and offer suggestions for the effective translation of cell spheroids for therapeutic treatments. [ABSTRACT FROM AUTHOR]