Mimicking bone microenvironment: 2D and 3D in vitro models of human osteoblasts

Yuste, Iván; Luciano, Francis C.; González-Burgos, Elena; Lalatsa, Aikaterini; Serrano, Dolores R.

doi:10.1016/j.phrs.2021.105626

Cited by 38 publications

(39 citation statements)

References 124 publications

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“…To make a good proximation to the real in vivo microenvironment, we examined the biological activities of SOOME in a 3D bioprinted module [ 46 ]. The SOOME 3D module was fabricated by our established PCL and cell-integrated 3D printing (PCI3D) system.…”

Section: Resultsmentioning

confidence: 99%

The Osteocyte Stimulated by Wnt Agonist SKL2001 Is a Safe Osteogenic Niche Improving Bioactivities in a Polycaprolactone and Cell Integrated 3D Module

Liu

Ruan

et al. 2022

Cells

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Finding and constructing an osteogenic microenvironment similar to natural bone tissue has always been a frontier topic in orthopedics. We found that osteocytes are targeting cells controlling bone anabolism produced by PTH (JBMR 2017, PMID: 27704638), and osteocytes with activated Wnt signaling orchestrate bone formation and resorption (PNAS 2015, PMID: 25605937). However, methods for taking advantage of the leading role of osteocytes in bone regeneration remain unexplored. Herein, we found that the osteocytes with SKL2001-activated Wnt signaling could be an osteogenic microenvironment (SOOME) which upregulates the expression of bone transcription factor Runx2 and Bglap and promotes the differentiation of bone marrow stromal cell ST2 into osteoblasts. Interestingly, 60 μM SKL2001 treatment of osteocytic MLO-Y4 for 24 h maintained Wnt signaling activation for three days after removal, which was sufficient to induce osteoblast differentiation. Triptonide, a Wnt inhibitor, could eliminate this differentiation. Moreover, on day 5, the Wnt signaling naturally decreased to the level of the control group, indicating that this method of Wnt-signaling induction is safe to use. We quickly verified in vivo function of SOOME to a good proximation in 3D bioprinted modules composed of reciprocally printed polycaprolactone bundles (for support) and cell bundles (for bioactivity). In the cell bundles, SOOME stably supported the growth and development of ST2 cells, the 7-day survival rate was as high as 91.6%, and proliferation ability increased linearly. Similarly, SOOME greatly promoted ST2 differentiation and mineralization for 28 days. In addition, SOOME upregulated the expression of angiopoietin 1, promoted endothelial cell migration and angiogenesis, and increased node number and total length of tubes and branches. Finally, we found that the function of SOOME could be realized through the paracrine pathway. This study reveals that osteocytes with Wnt signaling activated by SKL2001 are a safe osteogenic microenvironment. Both SOOME itself and its cell-free culture supernatant can improve bioactivity for osteoblast differentiation, with composite scaffolds especially bearing application value.

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Section: Resultsmentioning

confidence: 99%

The Osteocyte Stimulated by Wnt Agonist SKL2001 Is a Safe Osteogenic Niche Improving Bioactivities in a Polycaprolactone and Cell Integrated 3D Module

Liu

Ruan

et al. 2022

Cells

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“…The ECM allows the transmission of physical forces to the cell cytoskeleton via physical mechanotransduction, activating a signalling cascade, which affects cellular functions such as proliferation, migration, differentiation, and apoptosis ( Sikavitsas et al, 2001 ; Mccoy and O’brien, 2010 ; Wittkowske et al, 2016 ). Among these mechanical strains, fluid shear stress is induced by interstitial perfusion, which, in turn, results from pressure gradients produced by vascular and hydrostatic pressure, and ca be induced by mechanical loading ( Hillsley and Frangos, 1994 ; Mccoy and O’brien, 2010 ; Yao et al, 2012 ; Wittkowske et al, 2016 ; Yuste et al, 2021 ). These are all crucial players of mechanical stimulation of physiological tissue microenvironment ( Mccoy and O’brien, 2010 ; Wittkowske et al, 2016 ; Yuste et al, 2021 ) but also allow for increased diffusion of nutrients, metabolites, and oxygen, removal of toxic products or inhibitors of cellular metabolism, thereby preventing the formation of necrotic core areas ( Rouwkema et al, 2010 ; Place et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

“…Among these mechanical strains, fluid shear stress is induced by interstitial perfusion, which, in turn, results from pressure gradients produced by vascular and hydrostatic pressure, and ca be induced by mechanical loading ( Hillsley and Frangos, 1994 ; Mccoy and O’brien, 2010 ; Yao et al, 2012 ; Wittkowske et al, 2016 ; Yuste et al, 2021 ). These are all crucial players of mechanical stimulation of physiological tissue microenvironment ( Mccoy and O’brien, 2010 ; Wittkowske et al, 2016 ; Yuste et al, 2021 ) but also allow for increased diffusion of nutrients, metabolites, and oxygen, removal of toxic products or inhibitors of cellular metabolism, thereby preventing the formation of necrotic core areas ( Rouwkema et al, 2010 ; Place et al, 2017 ). Therefore, static in vitro cell cultures are highly limited by the lack of vasculature, which results in a scarce perfusion of cellular nutrients and dispersion of waste cellular products (200 µm).…”

Section: Introductionmentioning

confidence: 99%

“…To better resemble the bone microenvironment, several examples of 3D in vitro bone models are available, including the use of spheroids, 3D scaffolds, cell sheets, hydrogels, bioreactors, and microfluidics ( Wittkowske et al, 2016 ; Yuste et al, 2021 ). Among these, 3D in vitro fluidic macroscale (i.e., spinner flasks, rotating wall vessels), milliscale (customized perfusion bioreactors), and microscale systems (i.e., microfluidic devices) appear very promising to overcome the limitations of 2D cultures.…”

Section: Introductionmentioning

confidence: 99%

“…Among these, 3D in vitro fluidic macroscale (i.e., spinner flasks, rotating wall vessels), milliscale (customized perfusion bioreactors), and microscale systems (i.e., microfluidic devices) appear very promising to overcome the limitations of 2D cultures. These devices allow a fine tuning of dynamic interstitial perfusion ( Wittkowske et al, 2016 ; Yuste et al, 2021 ), a full understanding of cell-cell and cells-ECM interactions and, overall, a better comprehension of in vivo biological mechanisms ( Kim et al, 2007 ; Esch et al, 2015 ; Arrigoni et al, 2017 ; Carvalho et al, 2018 ; Wang et al, 2018 ; Nokhbatolfoghahaei et al, 2020a ). So far, good outcomes have been achieved in reproducing structural, functional, and mechanical properties of tissues using perfused platforms, including lung alveoli and bronchioles ( Huh et al, 2010 ; Ott et al, 2010 ; Price et al, 2010 ; Stucki et al, 2015 ), renal tubules and glomeruli ( Humes et al, 1999 ; Humes et al, 2004 ; Jang and Suh, 2010 ; Wilmer et al, 2016 ), small intestine ( Kimura et al, 2008 ; Imura et al, 2009 ; Pusch et al, 2011 ; Schweinlin et al, 2016 ), liver ( Kane et al, 2006 ; Tsang et al, 2007 ; Yamada et al, 2007 ; Domansky et al, 2010 ; Elbakary and Badhan, 2020 ) and the blood-brain barrier ( Booth and Kim, 2012 ; Griep et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

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Perfused Platforms to Mimic Bone Microenvironment at the Macro/Milli/Microscale: Pros and Cons

Lipreri

Baldini

Graziani

et al. 2022

Front. Cell Dev. Biol.

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As life expectancy increases, the population experiences progressive ageing. Ageing, in turn, is connected to an increase in bone-related diseases (i.e., osteoporosis and increased risk of fractures). Hence, the search for new approaches to study the occurrence of bone-related diseases and to develop new drugs for their prevention and treatment becomes more pressing. However, to date, a reliable in vitro model that can fully recapitulate the characteristics of bone tissue, either in physiological or altered conditions, is not available. Indeed, current methods for modelling normal and pathological bone are poor predictors of treatment outcomes in humans, as they fail to mimic the in vivo cellular microenvironment and tissue complexity. Bone, in fact, is a dynamic network including differently specialized cells and the extracellular matrix, constantly subjected to external and internal stimuli. To this regard, perfused vascularized models are a novel field of investigation that can offer a new technological approach to overcome the limitations of traditional cell culture methods. It allows the combination of perfusion, mechanical and biochemical stimuli, biological cues, biomaterials (mimicking the extracellular matrix of bone), and multiple cell types. This review will discuss macro, milli, and microscale perfused devices designed to model bone structure and microenvironment, focusing on the role of perfusion and encompassing different degrees of complexity. These devices are a very first, though promising, step for the development of 3D in vitro platforms for preclinical screening of novel anabolic or anti-catabolic therapeutic approaches to improve bone health.

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Levitational 3D Bioassembly and Density‐Based Spatial Coding of Levitoids

Moncal

Yaman

Durmus

2022

Adv Funct Materials

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Self‐assembly of cells into functional bioarchitectures with microscale spatial topographies are prevalent in nature. Despite the advances to recapitulate the native‐like microenvironment in vitro,fabrication of soft living materials with a deterministic control on the composition, functionality, and geometry, is still challenging. Here, a versatile, paramagnetically tunable, levitational biofabrication technique within a ring‐magnet system, RM‐LEV, is developed to assemble single cells or heterogeneous multicellular living architectures in a scaffold‐free manner. The self‐assembly and contactless biofabrication of multiple cell types into 3D multicellular “levitospheres” is demonstrated, where cellular positions are guided by levitation and density, simultaneously. Inherent density and diamagnetism of different cell types are leveraged to manipulate the geometry and self‐assembly of levitospheres into larger complex 3D structures, termed as “levitoids”. This approach is further applied to manipulate, position, and culture levitoids within a 3D hydrogel matrix under levitation, which preserves their viability and functionality. Thus, this study provides a foundation to create spatially heterogeneous 3D cellular assemblies with density‐coded bio‐architectures—which is not previously realized using magnetic levitation. This density‐based coding approach can be beneficial to study the cross‐talk between different cell types within spheroids and organoids, and be broadly applied to 3D bioprinting, tissue engineering, cancer, and neuroscience research.

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Mimicking bone microenvironment: 2D and 3D in vitro models of human osteoblasts

Cited by 38 publications

References 124 publications

The Osteocyte Stimulated by Wnt Agonist SKL2001 Is a Safe Osteogenic Niche Improving Bioactivities in a Polycaprolactone and Cell Integrated 3D Module

The Osteocyte Stimulated by Wnt Agonist SKL2001 Is a Safe Osteogenic Niche Improving Bioactivities in a Polycaprolactone and Cell Integrated 3D Module

Perfused Platforms to Mimic Bone Microenvironment at the Macro/Milli/Microscale: Pros and Cons

Levitational 3D Bioassembly and Density‐Based Spatial Coding of Levitoids

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