Microfluidic co-culture platform for investigating osteocyte-osteoclast signalling during fluid shear stress mechanostimulation

Middleton, Kevin; Al-Dujaili, Saja A.; X, Mei; Günther, Axel; You, Lidan

doi:10.1016/j.jbiomech.2017.05.012

Cited by 60 publications

(64 citation statements)

References 53 publications

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“…The use of compartmentalized culture environments promotes the selective application of those stimuli to different cell types. A 2D microfluidic platform with osteoclasts and osteocytes cultured in separate compartments was key to observe the cross-talk between mechanically stimulated osteocytes, osteoclast precursors and unstimulated osteocytes (You et al, 2008;Middleton et al, 2017). In general, microfabrication techniques applied to cell biology aims to develop advanced human disease models by the inclusion of pathological factors.…”

Section: Introductionmentioning

confidence: 99%

Primary Human Osteoblasts Cultured in a 3D Microenvironment Create a Unique Representative Model of Their Differentiation Into Osteocytes

Nasello

Alamán‐Díez

Schiavi

et al. 2020

Front. Bioeng. Biotechnol.

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Microengineered systems provide an in vitro strategy to explore the variability of individual patient response to tissue engineering products, since they prefer the use of primary cell sources representing the phenotype variability. Traditional in vitro systems already showed that primary human osteoblasts embedded in a 3D fibrous collagen matrix differentiate into osteocytes under specific conditions. Here, we hypothesized that translating this environment to the organ-on-a-chip scale creates a minimal functional unit to recapitulate osteoblast maturation toward osteocytes and matrix mineralization. Primary human osteoblasts were seeded in a type I collagen hydrogel, to establish the role of lower (2.5 × 10 5 cells/ml) and higher (1 × 10 6 cells/ml) cell density on their differentiation into osteocytes. A custom semi-automatic image analysis software was used to extract quantitative data on cellular morphology from brightfield images. The results are showing that cells cultured at a high density increase dendrite length over time, stop proliferating, exhibit dendritic morphology, upregulate alkaline phosphatase (ALP) activity, and express the osteocyte marker dental matrix protein 1 (DMP1). On the contrary, cells cultured at lower density proliferate over time, do not upregulate ALP and express the osteoblast marker bone sialoprotein 2 (BSP2) at all timepoints. Our work reveals that microengineered systems create unique conditions to capture the major aspects of osteoblast differentiation into osteocytes with a limited number of cells. We propose that the microengineered approach is a functional strategy to create a patient-specific bone tissue model and investigate the individual osteogenic potential of the patient bone cells.

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Section: Introductionmentioning

confidence: 99%

Primary Human Osteoblasts Cultured in a 3D Microenvironment Create a Unique Representative Model of Their Differentiation Into Osteocytes

Nasello

Alamán‐Díez

Schiavi

et al. 2020

Front. Bioeng. Biotechnol.

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“…For experiments, the device was placed into a manifold (Fig. B) using techniques previously reported . A detailed description of this setup can be found in the supplemental methods.…”

Section: Methodsmentioning

confidence: 99%

“…1B) using techniques previously reported. 23 A detailed description of this setup can be found in the supplemental methods. MLO-Y4 cells were then seeded in the device at a density of 1 Â 10 6 cells/ml and allowed to attach for 2 h at 37˚C in an incubator.…”

Section: Experimental Device Preparationmentioning

confidence: 99%

Microfluidics approach to investigate the role of dynamic similitude in osteocyte mechanobiology

Middleton

Kondiboyina

Borrett

et al. 2017

Journal Orthopaedic Research

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Fluid flow is an important regulator of cell function and metabolism in many tissues. Fluid shear stresses have been used to level the mechanical stimuli applied in vitro with what occurs in vivo. However, these experiments often lack dynamic similarity, which is necessary to ensure the validity of the model. For interstitial fluid flow, the major requirement for dynamic similarity is the Reynolds number (Re), the ratio of inertial to viscous forces, is the same between the system and model. To study the necessity of dynamic similarity for cell mechanotransduction studies, we investigated the response of osteocyte-like MLO-Y4 cells to different Re flows at the same level of fluid shear stress. Osteocytes were chosen for this study as flows applied in vitro and in vivo have Re that are orders of magnitude different. We hypothesize that osteocytes' response to fluid flow is Re dependent. We observed that cells exposed to lower and higher Re flows developed rounded and triangular morphologies, respectively. Lower Re flows also reduced apoptosis rates compared to higher Re flows. Furthermore, MLO-Y4 cells exposed to higher Re flows had stronger calcium responses compared to lower Re flows. However, by also controlling for flow rate, the lower Re flows induced a stronger calcium response; while degradation of components of the osteocyte glycocalyx reversed this effect. This work suggests that osteocytes are highly sensitive to differences in Re, independent of just shear stresses, supporting the need for improved in vitro flow platforms that better recapitulate the physiological environment. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:663-671, 2018.

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“…Furthermore, microfluidic devices have also been used outside of endothelial research to record phenotypic transformations of aortic valve interstitial cells during applied shear stress and to monitor morphological changes of osteocytes during application of flow. Additionally, Middleton et al (2017a) showed that in the presence of shear stress, cell to cell interactions of osteocytes co-cultured with osteoclast are enhanced, leading to the improved mechanical response of bone cells. A more detailed review on monitoring cell-cell interaction using microfluidic devices was recently published by Rothbauer et al (2017) .…”

Section: Microfluidic Mechanobiology In Monolayers and Barrier Modelsmentioning

confidence: 99%

Small Force, Big Impact: Next Generation Organ-on-a-Chip Systems Incorporating Biomechanical Cues

et al. 2018

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Mechanobiology-on-a-chip is a growing field focusing on how mechanical inputs modulate physico-chemical output in microphysiological systems. It is well known that biomechanical cues trigger a variety of molecular events and adjustment of mechanical forces is therefore essential for mimicking in vivo physiologies in organ-on-a-chip technology. Biomechanical inputs in organ-on-a-chip systems can range from variations in extracellular matrix type and stiffness and applied shear stresses to active stretch/strain or compression forces using integrated flexible membranes. The main advantages of these organ-on-a-chip systems are therefore (a) the control over spatiotemporal organization of in vivo-like tissue architectures, (b) the ability to precisely control the amount, duration and intensity of the biomechanical stimuli, and (c) the capability of monitoring in real time the effects of applied mechanical forces on cell, tissue and organ functions. Consequently, over the last decade a variety of microfluidic devices have been introduced to recreate physiological microenvironments that also account for the influence of physical forces on biological functions. In this review we present recent advances in mechanobiological lab-on-a-chip systems and report on lessons learned from these current mechanobiological models. Additionally, future developments needed to engineer next-generation physiological and pathological organ-on-a-chip models are discussed.

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Microfluidic co-culture platform for investigating osteocyte-osteoclast signalling during fluid shear stress mechanostimulation

Cited by 60 publications

References 53 publications

Primary Human Osteoblasts Cultured in a 3D Microenvironment Create a Unique Representative Model of Their Differentiation Into Osteocytes

Primary Human Osteoblasts Cultured in a 3D Microenvironment Create a Unique Representative Model of Their Differentiation Into Osteocytes

Microfluidics approach to investigate the role of dynamic similitude in osteocyte mechanobiology

Small Force, Big Impact: Next Generation Organ-on-a-Chip Systems Incorporating Biomechanical Cues

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