A Three-Dimensional Mechanical Loading Model of Human Osteocytes in Their Native Matrix

Zhang, Chen; Farré-Guasch, Elisabet; Jin, Jianfeng; Essen, H.W. van; Klein‐Nulend, Jenneke; Bravenboer, Nathalie

doi:10.1007/s00223-021-00919-z

Cited by 4 publications

(14 citation statements)

References 44 publications

(82 reference statements)

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“…Osteocyte viability and sclerostin expression are similar immediately after isolation and culture up to 2 days. ( 48 ) Thus, osteocyte mechanoresponsiveness is unlikely to be changed by simply putting the bone pieces in culture. Explants were either not loaded or mechanically loaded by three‐point bending with a rounded contact point and a span length of 8 mm, at a magnitude of 2000 or 8000 μɛ at 1 Hz frequency for 5 minutes, as described.…”

Section: Methodsmentioning

confidence: 99%

“…Explants were either not loaded or mechanically loaded by three‐point bending with a rounded contact point and a span length of 8 mm, at a magnitude of 2000 or 8000 μɛ at 1 Hz frequency for 5 minutes, as described. ( 48 ) In short, mechanical loading was applied using a custom‐made loading apparatus, which generated sinusoidal displacement using a computer‐driven voice coil linear microactuator (type NCM04‐25250‐2LVE; H2W Technologies, Valencia, CA, USA). ( 48 ) Mechanical loading at 2000 μɛ mimics physiological loading, ( 49 , 50 ) and 8000 μɛ mimics pathological overloading.…”

Section: Methodsmentioning

confidence: 99%

“…( 48 ) In short, mechanical loading was applied using a custom‐made loading apparatus, which generated sinusoidal displacement using a computer‐driven voice coil linear microactuator (type NCM04‐25250‐2LVE; H2W Technologies, Valencia, CA, USA). ( 48 ) Mechanical loading at 2000 μɛ mimics physiological loading, ( 49 , 50 ) and 8000 μɛ mimics pathological overloading. ( 51 ) The osteocytes in bone explants exposed to mechanical loading at 1600 μɛ are highly viable at 24 hours post‐culture, as shown in our previous study.…”

Section: Methodsmentioning

confidence: 99%

“…( 48 ) The osteocytes in the loaded bone explants keep their specific phenotype, and are able to respond to mechanical loading. ( 48 ) This 3D‐mechanical loading model of bone explants in vitro mimics osteocytes in their native microenvironment in vivo. ( 48 ) To date, the mechanism of osteocyte‐orchestrated adaptation of bone to mechanical loading is still not fully understood.…”

Section: Introductionmentioning

confidence: 99%

“…( 48 ) This 3D‐mechanical loading model of bone explants in vitro mimics osteocytes in their native microenvironment in vivo. ( 48 ) To date, the mechanism of osteocyte‐orchestrated adaptation of bone to mechanical loading is still not fully understood. Therefore, more insight into target gene expression by osteocytes in their native matrix in response to mechanical loading is necessary.…”

Section: Introductionmentioning

confidence: 99%

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Mapping the Response of Human Osteocytes in Native Matrix to Mechanical Loading Using RNA Sequencing

et al. 2023

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Osteocytes sense mechanical loads and transduce mechanical signals into a chemical response. They are the most abundant bone cells deeply embedded in mineralized bone matrix, which affects their regulatory activity in the mechanical adaptation of bone. The specific location in the calcified bone matrix hinders studies on osteocytes in the in vivo setting. Recently, we developed a three‐dimensional mechanical loading model of human osteocytes in their native matrix, allowing to study osteocyte mechanoresponsive target gene expression in vitro. Here we aimed to identify differentially expressed genes by mapping the response of human primary osteocytes in their native matrix to mechanical loading using RNA sequencing. Human fibular bone was retrieved from 10 donors (age: 32–82 years, 5 female, 5 male). Cortical bone explants (8.0 × 3.0 × 1.5 mm; length × width × height) were either not loaded or mechanically loaded by 2000 or 8000 μɛ for 5 minutes, followed by 0, 6, or 24 hours post‐culture without loading. High‐quality RNA was isolated, and differential gene expression analysis performed by R2 platform. Real‐time PCR was used to confirm differentially expressed genes. Twenty‐eight genes were differentially expressed between unloaded and loaded (2000 or 8000 μɛ) bone at 6 hours post‐culture, and 19 genes at 24 hours post‐culture. Eleven of these genes were related to bone metabolism, ie, EGR1 , FAF1 , H3F3B , PAN2 , RNF213 , SAMD4A , and TBC1D24 at 6 hours post‐culture, and EGFEM1P , HOXD4 , SNORD91B , and SNX9 at 24 hours post‐culture. Mechanical loading significantly decreased RNF213 gene expression, which was confirmed by real‐time PCR. In conclusion, mechanically loaded osteocytes differentially expressed 47 genes, of which 11 genes were related to bone metabolism. RNF213 might play a role in mechanical adaptation of bone by regulating angiogenesis, which is a prerequisite for successful bone formation. The functional aspects of the differentially expressed genes in bone mechanical adaptation requires future investigation. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mapping the Response of Human Osteocytes in Native Matrix to Mechanical Loading Using RNA Sequencing

et al. 2023

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The Impact of Culture Variables on a 3D Human In Vitro Bone Remodeling Model: A Design of Experiments Approach

Wildt

Cuypers

Cramer

et al. 2023

Adv Healthcare Materials

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Human in vitro bone remodeling models, using osteoclast–osteoblast cocultures, can facilitate the investigation of human bone remodeling while reducing the need for animal experiments. Although current in vitro osteoclast–osteoblast cocultures have improved the understanding of bone remodeling, it is still unknown which culture conditions support both cell types. Therefore, in vitro bone remodeling models can benefit from a thorough evaluation of the impact of culture variables on bone turnover outcomes, with the aim to reach balanced osteoclast and osteoblast activity, mimicking healthy bone remodeling. Using a resolution III fractional factorial design, the main effects of commonly used culture variables on bone turnover markers in an in vitro human bone remodeling model are identified. This model is able to capture physiological quantitative resorption–formation coupling along all conditions. Culture conditions of two runs show promising results: conditions of one run can be used as a high bone turnover system and conditions of another run as a self‐regulating system as the addition of osteoclastic and osteogenic differentiation factors is not required for remodeling. The results generated with this in vitro model allow for better translation between in vitro studies and in vivo studies, toward improved preclinical bone remodeling drug development.

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Titanium Nanotopography Improves Mechano-Response of Osteocyte Three-Dimensional Network Enhancing Bone Turnover Capability

He,

Yamada,

Watanabe

et al. 2023

Preprint

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A Three-Dimensional Mechanical Loading Model of Human Osteocytes in Their Native Matrix

Cited by 4 publications

References 44 publications

Mapping the Response of Human Osteocytes in Native Matrix to Mechanical Loading Using RNA Sequencing

Mapping the Response of Human Osteocytes in Native Matrix to Mechanical Loading Using RNA Sequencing

The Impact of Culture Variables on a 3D Human In Vitro Bone Remodeling Model: A Design of Experiments Approach

Titanium Nanotopography Improves Mechano-Response of Osteocyte Three-Dimensional Network Enhancing Bone Turnover Capability

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