RRC ID 59663
Author Sato K, Matsubara O, Hase E, Minamikawa T, Yasui T.
Title Quantitative in situ time-series evaluation of osteoblastic collagen synthesis under cyclic strain using second-harmonic-generation microscopy.
Journal J Biomed Opt
Abstract The aim of this study is to evaluate the osteoblastic collagen synthesis under mechanical stimulation using second-harmonic-generation (SHG) microscopy. We apply SHG microscopy to monitor the collagen fibers synthesized by osteoblast-like cells (MC3T3-E1) without the need for fixation and staining. To quantitatively evaluate the influence of mechanical stimulation on osteoblastic collagen synthesis, we compare SHG images of osteoblast-synthesized collagen fibers with and without a cyclic stretch stimulus applied using a lab-made stretching device. We acquire SHG images every 7 days for 3 weeks at different stimulus conditions (5 min/day and 3 h/day with a strain magnitude of 5% and a frequency of 0.5 Hz). Image analysis of the average SHG intensity indicates that the amount of osteoblastic collagen synthesis is significantly enhanced by the cyclic stretch compared with the nonstretched condition, while there is no significant difference between the two mechanical stimulation conditions. Furthermore, the maturity of the collagen fibers in the early stage of bone formation is not affected by the mechanical stimulation. The results can be used in bone regenerative medicine to apply feedback control of collagen synthesis by artificial stimulation.
Volume 24(3)
Pages 1-8
Published 2019-1-1
DOI 10.1117/1.JBO.24.3.031019
PII JBO-180368SSRR
PMID 30635995
PMC PMC6975189
MeSH Collagen / analysis* Collagen / physiology Humans Image Interpretation, Computer-Assisted / methods* Microscopy / methods* Stress, Mechanical
IF 2.555
Times Cited 0
Resource
Human and Animal Cells MC3T3-E1(RCB1126)