TY - JOUR
T1 - Mapping the Response of Human Osteocytes in Native Matrix to Mechanical Loading Using RNA Sequencing
AU - Zhang, Chen
AU - van Essen, Huib W.
AU - Sie, Daoud
AU - Micha, Dimitra
AU - Pals, Gerard
AU - Klein-Nulend, Jenneke
AU - Bravenboer, Nathalie
N1 - Funding Information: We thank Behrouz Zandieh‐Doulabi for designing the primers, and thank Richard Volckmann for advice on the RNA‐seq data analysis using R2 platform. This work was granted by the China Scholarship Council (CSC, No. 201706320330), Amsterdam Movement Sciences (AMS Innovation Grant 2017), and Health‐Holland (Project No. LSHM19016, “BB”). Funding Information: We thank Behrouz Zandieh-Doulabi for designing the primers, and thank Richard Volckmann for advice on the RNA-seq data analysis using R2 platform. This work was granted by the China Scholarship Council (CSC, No. 201706320330), Amsterdam Movement Sciences (AMS Innovation Grant 2017), and Health-Holland (Project No. LSHM19016, “BB”). Publisher Copyright: © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.
PY - 2023/4
Y1 - 2023/4
N2 - 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.
AB - 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.
KW - 3D
KW - HUMAN BONE
KW - MECHANICAL LOADING
KW - NATIVE MATRIX
KW - OSTEOCYTES
KW - RNA SEQUENCING
UR - http://www.scopus.com/inward/record.url?scp=85148506747&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85148506747&partnerID=8YFLogxK
U2 - https://doi.org/10.1002/jbm4.10721
DO - https://doi.org/10.1002/jbm4.10721
M3 - Article
C2 - 37065632
SN - 2473-4039
VL - 7
SP - 1
EP - 16
JO - JBMR Plus
JF - JBMR Plus
IS - 4
M1 - e10721
ER -