Differential behaviour and gene expression in 3D cultures of femoral‐ and calvarial‐derived human osteoblasts under a cyclic compressive mechanical load

2021 ◽  
Author(s):  
Yana Itskovich ◽  
Murray C Meikle ◽  
Richard D Cannon ◽  
Mauro Farella ◽  
Dawn E Coates ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mechanical Load ◽  
Human Osteoblasts ◽  
3D Cultures ◽  
Differential Behaviour

scholarly journals Adhesion Behaviour of Primary Human Osteoblasts and Fibroblasts on Polyether Ether Ketone Compared with Titanium under In Vitro Lipopolysaccharide Incubation

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2739 ◽  
Author(s):  
Korbinian Benz ◽  
Andreas Schöbel ◽  
Marisa Dietz ◽  
Peter Maurer ◽  
Jochen Jackowski
Keyword(s):  
Gene Expression ◽  
Protein Level ◽  
Binding Protein ◽  
Sem Images ◽  
Human Osteoblasts ◽  
Polyether Ether Ketone ◽  
Primary Human Osteoblasts ◽  
Titanium Surfaces ◽  
Ether Ketone

The aim of this in vitro pilot study was to analyse the adhesion behaviour of human osteoblasts and fibroblasts on polyether ether ketone (PEEK) when compared with titanium surfaces in an inflammatory environment under lipopolysaccharide (LPS) incubation. Scanning electron microscopy (SEM) images of primary human osteoblasts/fibroblasts on titanium/PEEK samples were created. The gene expression of the LPS-binding protein (LBP) and the LPS receptor (toll-like receptor 4; TLR4) was measured by real-time polymerase chain reaction (PCR). Immunocytochemistry was used to obtain evidence for the distribution of LBP/TLR4 at the protein level of the extra-cellular-matrix-binding protein vinculin and the actin cytoskeleton. SEM images revealed that the osteoblasts and fibroblasts on the PEEK surfaces had adhesion characteristics comparable to those of titanium. The osteoblasts contracted under LPS incubation and a significantly increased LBP gene expression were detected. This was discernible at the protein level on all the materials. Whereas no increase of TLR4 was detected with regard to mRNA concentrations, a considerable increase in the antibody reaction was detected on all the materials. As is the case with titanium, the colonisation of human osteoblasts and fibroblasts on PEEK samples is possible under pro-inflammatory environmental conditions and the cellular inflammation behaviour towards PEEK is lower than that of titanium.

The effect of shock wave therapy on gene expression in human osteoblasts isolated from hypertrophic fracture non-unions

Shock Waves ◽  
2014 ◽  
Vol 25 (1) ◽  
pp. 91-102 ◽  
Author(s):  
A. Hofmann ◽  
U. Ritz ◽  
J.-D. Rompe ◽  
A. Tresch ◽  
P. M. Rommens
Keyword(s):  
Gene Expression ◽  
Shock Wave ◽  
Shock Wave Therapy ◽  
Human Osteoblasts

Gene expression, cytoskeletal changes and extracellular matrix synthesis in human osteoblasts treated with cyclosporin A

2009 ◽  
Vol 63 (9) ◽  
pp. 619-626 ◽  
Author(s):  
Maurizio Vertemati ◽  
Ernesto Minola ◽  
Claudia Dolci ◽  
Giordano Stabellini ◽  
Furio Pezzetti ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Cyclosporin A ◽  
Matrix Synthesis ◽  
Human Osteoblasts

Integrin signaling's potential for mediating gene expression in hypertrophying skeletal muscle

2000 ◽  
Vol 88 (1) ◽  
pp. 337-343 ◽  
Author(s):  
James A. Carson ◽  
Lei Wei
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Signaling Pathway ◽  
Mechanical Load ◽  
Integrin Signaling ◽  
Mechanical Forces ◽  
Growth Factor Signaling ◽  
Muscle Gene

Overloaded skeletal muscle undergoes dramatic shifts in gene expression, which alter both the phenotype and mass. Molecular biology techniques employing both in vivo and in vitro hypertrophy models have demonstrated that mechanical forces can alter skeletal muscle gene regulation. This review's purpose is to support integrin-mediated signaling as a candidate for mechanical load-induced hypertrophy. Research quantifying components of the integrin-signaling pathway in overloaded skeletal muscle have been integrated with knowledge regarding integrins role during development and cardiac hypertrophy, with the hope of demonstrating the pathway's importance. The role of integrin signaling as an integrator of mechanical forces and growth factor signaling during hypertrophy is discussed. Specific components of integrin signaling, including focal adhesion kinase and low-molecular-weight GTPase Rho are mentioned as downstream targets of this signaling pathway. There is a need for additional mechanistic studies capable of providing a stronger linkage between integrin-mediated signaling and skeletal muscle hypertrophy; however, there appears to be abundant justification for this type of research.

Extracorporeal Shock Wave-Mediated Changes in Proliferation, Differentiation, and Gene Expression of Human Osteoblasts

2008 ◽  
Vol 65 (6) ◽  
pp. 1402-1410 ◽  
Author(s):  
Alexander Hofmann ◽  
Ulrike Ritz ◽  
Martin Henri Hessmann ◽  
Mauro Alini ◽  
Pol Maria Rommens ◽  
...  
Keyword(s):  
Gene Expression ◽  
Shock Wave ◽  
Human Osteoblasts ◽  
Extracorporeal Shock Wave

scholarly journals Role of PDK1 in skeletal muscle hypertrophy induced by mechanical load

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Naoki Kuramoto ◽  
Kazuhiro Nomura ◽  
Daisuke Kohno ◽  
Tadahiro Kitamura ◽  
Gerard Karsenty ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Muscle Mass ◽  
Skeletal Muscle Mass ◽  
Muscle Hypertrophy ◽  
Mechanical Load ◽  
Static Condition ◽  
Ribosomal Protein S6 ◽  
Pi3k Signaling Pathway ◽  
Induced Changes

AbstractPhosphatidylinositol 3-kinase (PI3K) plays an important role in protein metabolism and cell growth. We here show that mice (M-PDK1KO mice) with skeletal muscle–specific deficiency of 3′-phosphoinositide–dependent kinase 1 (PDK1), a key component of PI3K signaling pathway, manifest a reduced skeletal muscle mass under the static condition as well as impairment of mechanical load–induced muscle hypertrophy. Whereas mechanical load-induced changes in gene expression were not affected, the phosphorylation of ribosomal protein S6 kinase (S6K) and S6 induced by mechanical load was attenuated in skeletal muscle of M-PDK1KO mice, suggesting that PDK1 regulates muscle hypertrophy not through changes in gene expression but through stimulation of kinase cascades such as the S6K-S6 axis, which plays a key role in protein synthesis. Administration of the β2-adrenergic receptor (AR) agonist clenbuterol activated the S6K-S6 axis in skeletal muscle and induced muscle hypertrophy in mice. These effects of clenbuterol were attenuated in M-PDK1KO mice, and mechanical load–induced activation of the S6K-S6 axis and muscle hypertrophy were inhibited in mice with skeletal muscle–specific deficiency of β2-AR. Our results suggest that PDK1 regulates skeletal muscle mass under the static condition and that it contributes to mechanical load–induced muscle hypertrophy, at least in part by mediating signaling from β2-AR.

scholarly journals Different Cyclic Mechanical Loading Patterns Alter Expression of Osteogenic Markers in a Fibroblast-Osteoblast Co-Culture Model Resembling the Tendon-To-Bone Interface After Autologous ACL Reconstruction

2014 ◽  
Vol 2 (11_suppl3) ◽  
pp. 2325967114S0019
Author(s):  
Philip Peter Roessler ◽  
Johannes Struewer ◽  
Jürgen Rudolph Paletta ◽  
Turgay Efe
Keyword(s):  
Gene Expression ◽  
Mechanical Loading ◽  
Mechanical Load ◽  
Cruciate Ligament ◽  
Culture Conditions ◽  
Bone Morphogenetic Protein 2 ◽  
Marker Genes ◽  
Bone Interface ◽  
Loading Patterns

Objectives: The aim of the study was to evaluate the influence of different cyclical mechanical loading patterns on co-cultures of fibroblasts and osteoblasts in vitro, simulating the conditions of the tendon-to-bone interface after anterior cruciate ligament reconstruction. Methods: Tendon-derived rodent fibroblasts (TDF) and osteoblast-like cells (OBL) were co-cultured to simulate the tendon-to-bone interface. Cyclical loading was applied for one hour twice a day for three days, with a frequency of 1 Hz and 3 % strain. Alkaline phosphatase (AP), osteocalcin (OC), collagen type 1 (COL1A1), and bone morphogenetic protein 2 (BMP-2) gene expression and protein deposition were detected by real-time polymerase chain reaction (qPCR) and immunocytochemical analysis. Results: Mechanical loading significantly decreased AP, OC, and COL1A1 gene expression in both OBL and TDF, compared to non-loaded cultures. However, mechanical load increased gene expression of the same marker genes including BMP-2 during co-culture. Immunocytochemistry demonstrated increased deposition of corresponding proteins in the same range, independent of culture conditions. Higher depositions of BMP-2 were shown under loading conditions for osteoblast and TDF monocultures. Prolongation of mechanical loading resulted in cell detachment and spheroid formation. Conclusion: Cyclical mechanical loading caused downregulation of genes involved in osteointegration and osteoinduction, such as OC, ALP, and COL1A1 in monocultures of osteoblasts and fibroblasts; co-cultures lacked this phenomenon. Immunocytochemistry and qPCR analysis showed slight upregulations of marker genes and corresponding proteins. This might be due to the potential stabilising effects of osteoblast-fibroblast cross talk in the co-culture environment, resembling fibrocartilage formation at the tendon-to-bone interface.

Microarray-based gene expression analysis of human osteoblasts in response to different biomaterials

2009 ◽  
Vol 88A (2) ◽  
pp. 401-408 ◽  
Author(s):  
Karina F. Bombonato-Prado ◽  
Larissa S. Bellesini ◽  
Cristina M. Junta ◽  
Márcia M. Marques ◽  
Geraldo A. Passos ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Human Osteoblasts
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