Osteoblast differentiation is enhanced in rotary cell culture simulated microgravity environments

Y Joon Ko; Rebecca S Zaharias; Denise A Seabold; John Lafoon; Galen B Schneider

doi:10.1111/j.1532-849X.2007.00204.x

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Osteoblast differentiation is enhanced in rotary cell culture simulated microgravity environments

Journal article

Peer reviewed

Osteoblast differentiation is enhanced in rotary cell culture simulated microgravity environments

Y Joon Ko, Rebecca S Zaharias, Denise A Seabold, John Lafoon and Galen B Schneider

Journal of prosthodontics, Vol.16(6), pp.431-438

11/2007

DOI: 10.1111/j.1532-849X.2007.00204.x

PMID: 17559538

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Abstract

As the aging population increases, more people will become reliant on regenerative dental medicine for implant therapy. The objective of this study was to test the hypothesis that 3D rotary cell culture (RCC) environments created by simulated microgravity would enhance osteogenic gene expression using integrin mediated pathways. Human embryonic palatal mesenchymal (HEPM, ATCC 1486) pre-osteoblasts were cultured in either RCC to create 3D environments or in 2D monolayers for 72 hours. Gross phenotypic analysis was performed using Alizarin Red S staining for calcium and microscopy. Real-time PCR analysis was used to detect differences in osteoblast gene expression. Aggregates developed in 3D RCC environments were treated with or without antibody to the collagen-I integrin receptor alpha2beta1 to determine whether this molecular pathway might contribute to the development of a mineralized matrix. Microscopic analysis demonstrated that RCC environments promoted 3D aggregate formation by 72 hours without any scaffold. The mass appeared osseous-like with a white, shiny, translucent surface. The center was amorphous with areas of vacuolization, tubule-like structures, and fibrous-like extensions. Real-time PCR data showed that 3D environments enhanced osteogenic gene expression as compared with 2D monolayer culturing conditions. At 72 hours, changes in levels of osteogenic gene expression were noted. Cbfa1, a necessary transcription factor for osteoblast differentiation, was expressed 33% higher (p= 0.26); Collagen 1, 69% higher (p= 0.05); Osterix, 49% higher (p= 0.001); and BSPII, 54% higher (p= 0.001) than osteoblasts cultured for 72 hours in standard 2D monolayer conditions. When cultured in the presence of collagen alpha2beta1 integrin receptor antibody, 3D aggregates had decreased levels of mineralization as compared with non-treated aggregates. RCC enhances osteoblast differentiation using integrin mediated pathways.

Calcification, Physiologic - genetics

Osteogenesis - physiology

Cell Proliferation

Integrin-Binding Sialoprotein

Coloring Agents

Humans

Osteoblasts - physiology

Cells, Cultured

Core Binding Factor Alpha 1 Subunit - analysis

Calcium - analysis

Mesoderm - cytology

Rotation

Phenotype

Integrin alpha2beta1 - analysis

Anthraquinones

Cell Adhesion - physiology

Polymerase Chain Reaction

Sp7 Transcription Factor

Cell Differentiation

Collagen Type I - analysis

Transcription Factors - analysis

Weightlessness Simulation

Sialoglycoproteins - analysis

Details

Title: Subtitle: Osteoblast differentiation is enhanced in rotary cell culture simulated microgravity environments
Creators: Y Joon Ko - Department of Prosthodontics, College of Dentistry, University of Iowa, Iowa City, IA, USA
Rebecca S Zaharias
Denise A Seabold
John Lafoon
Galen B Schneider
Resource Type: Journal article
Publication Details: Journal of prosthodontics, Vol.16(6), pp.431-438
Publisher: United States
DOI: 10.1111/j.1532-849X.2007.00204.x
PMID: 17559538
ISSN: 1059-941X
eISSN: 1532-849X
Grant note: R21-DE016677-01 / NIDCR NIH HHS P60-DE13076 / NIDCR NIH HHS
Language: English
Date published: 11/2007
Academic Unit: Anatomy and Cell Biology; Dentistry Administration; Prosthodontics
Record Identifier: 9984066089202771

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