Isolation of osteocytes from human trabecular bone

Prideaux, Matthew; Schutz, Christine; Wijenayaka, Asiri R.; Findlay, David M.; Campbell, D.; Solomon, Lucian B.; Atkins, Gerald J.

doi:10.1016/j.bone.2016.04.017

Cited by 36 publications

(36 citation statements)

References 35 publications

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“…Prideaux et al[35] recently reported that they could isolated human osteocytes using 5 digestion cycles or more (<8 cycles) and reported similar results to ours. This study however focused only on the isolation and did not track the cell population change during 2D culture.…”

Section: Discussionsupporting

confidence: 86%

“…The proliferation of bone cells can lead to the formation of disordered and random cell aggregates, as reported by: (1) Prideaux et al[35] for human primary osteocyte culture in collagen gel, Mulcahy et al[36] and (2) Vazquez et al[37] for MLO-Y4 cell cultures with matrigel and collagen gel, respectively. These 3D gel culture approaches resulted in very low cell densities and physiologically irrelevant cell-to-cell distance and 3D network structure.…”

Section: Discussionmentioning

confidence: 99%

“…Since osteocytes have a typical diameter of 8–10 µm, microbeads with diameters of 20~25 µm were chosen to allow a cell to be placed within the interstitial site that is sufficiently large, but small for occupation by more than a few cells. Compared to other state-of-the-art bone tissue models,[35]–[39] our approach uniquely replicates physiologically relevant cell-to-cell distance, phenotype, and functions.…”

Section: Introductionmentioning

confidence: 99%

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Ex vivo construction of human primary 3D–networked osteocytes

Sun

Choudhary

Mannion

et al. 2017

Bone

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A human bone tissue model was developed by constructing ex vivo the 3D network of osteocytes via the biomimetic assembly of primary human osteoblastic cells with 20–25 µm microbeads and subsequent microfluidic perfusion culture. The biomimetic assembly: (1) enabled 3D-constructed cells to form cellular network via processes with an average cell-to-cell distance of 20–25 µm, and (2) inhibited cell proliferation within the interstitial confine between the microbeads while the confined cells produced extracellular matrix (ECM) to form a mechanically integrated structure. The mature osteocytic expressions of SOST and FGF23 genes became significantly higher, especially for SOST by 250 folds during 3D culture. The results validate that the bone tissue model: (1) consists of 3D cellular network of primary human osteocytes, (2) mitigates the osteoblastic differentiation and proliferation of primary osteoblast-like cells encountered in 2D culture, and (3) therefore reproduces ex vivo the phenotype of human 3D-networked osteocytes. The 3D tissue construction approach is expected to provide a clinically relevant and high-throughput means for evaluating drugs and treatments that target bone diseases with in vitro convenience.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ex vivo construction of human primary 3D–networked osteocytes

Sun

Choudhary

Mannion

et al. 2017

Bone

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“…Bone cells can be sourced from different origins including pluripotent stem cells which are differentiating to osteoblasts (Jaiswal et al, 1997),primary osteoblasts and osteocytes from different species (Jonsson et al, 2009; Prideaux et al, 2016), andimmortalized and osteosarcoma cell lines (Kato et al, 2001; Woo et al, 2011). …”

Section: Bone Mechanotransductionmentioning

confidence: 99%

“…primary osteoblasts and osteocytes from different species (Jonsson et al, 2009; Prideaux et al, 2016), and…”

Section: Bone Mechanotransductionmentioning

confidence: 99%

In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

Wittkowske

Reilly

Lacroix

et al. 2016

Front. Bioeng. Biotechnol.

233

225

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This review describes the role of bone cells and their surrounding matrix in maintaining bone strength through the process of bone remodeling. Subsequently, this work focusses on how bone formation is guided by mechanical forces and fluid shear stress in particular. It has been demonstrated that mechanical stimulation is an important regulator of bone metabolism. Shear stress generated by interstitial fluid flow in the lacunar-canalicular network influences maintenance and healing of bone tissue. Fluid flow is primarily caused by compressive loading of bone as a result of physical activity. Changes in loading, e.g., due to extended periods of bed rest or microgravity in space are associated with altered bone remodeling and formation in vivo. In vitro, it has been reported that bone cells respond to fluid shear stress by releasing osteogenic signaling factors, such as nitric oxide, and prostaglandins. This work focusses on the application of in vitro models to study the effects of fluid flow on bone cell signaling, collagen deposition, and matrix mineralization. Particular attention is given to in vitro set-ups, which allow long-term cell culture and the application of low fluid shear stress. In addition, this review explores what mechanisms influence the orientation of collagen fibers, which determine the anisotropic properties of bone. A better understanding of these mechanisms could facilitate the design of improved tissue-engineered bone implants or more effective bone disease models.

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Cathepsin K Controls Cortical Bone Formation by Degrading Periostin

Bonnet

Brun

Rousseau

et al. 2017

Journal of Bone and Mineral Research

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Although inhibitors of bone resorption concomitantly reduce bone formation because of the coupling between osteoclasts and osteoblasts, inhibition or deletion of cathepsin k (CatK) stimulates bone formation despite decreasing resorption. The molecular mechanisms responsible for this increase in bone formation, particularly at periosteal surfaces where osteoclasts are relatively poor, remain unclear. Here we show that CatK pharmacological inhibition or deletion (Ctsk mice) potentiates mechanotransduction signals mediating cortical bone formation. We identify periostin (Postn) as a direct molecular target for degradation by CatK and show that CatK deletion increases Postn and β-catenin expression in vivo, particularly at the periosteum. In turn, Postn deletion selectively abolishes cortical, but not trabecular, bone formation in CatK-deficient mice. Taken together, these data indicate that CatK not only plays a major role in bone remodeling but also modulates modeling-based cortical bone formation by degrading periostin and thereby moderating Wnt-β-catenin signaling. These findings provide novel insights into the role of CatK on bone homeostasis and the mechanisms of increased cortical bone volume with CatK mutations and pharmacological inhibitors. © 2017 American Society for Bone and Mineral Research.

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Isolation of osteocytes from human trabecular bone

Cited by 36 publications

References 35 publications

Ex vivo construction of human primary 3D–networked osteocytes

Ex vivo construction of human primary 3D–networked osteocytes

In Vitro Bone Cell Models: Impact of Fluid Shear Stress on Bone Formation

Cathepsin K Controls Cortical Bone Formation by Degrading Periostin

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