PhD Exit Seminar – Roxanne Fournier (Harrison lab)

Osteocyte Form and Function in Loading and Unloading Environments

Wednesday, October 14^th, 2020 at 1:10pm 

Join Zoom Meeting:

https://us02web.zoom.us/j/86106153621?pwd=WVhhUUJyZFJNc3JRVzAxd0d3d2JCUT09

Meeting ID: 861 0615 3621

Passcode: Bonecell

Abstract: 

Long-term human spaceflight presents unprecedented challenges to the human body, such as microgravity-induced bone loss. Without intervention, astronauts lose between 1 – 2% bone mineral density per month in weight bearing bones and the cellular mechanisms involved in this process are still poorly understood. To date, the most abundant cell of bone, the osteocyte, has rarely been subjected to spaceflight. Yet osteocytes are known to be important regulators of bone loss in immobilized and elderly people. The first aim of this thesis was to develop novel 3D culture methods to study osteocytes using the Rotary Cell Culture System (RCCS). This device is commonly used for both suspension culture and simulated microgravity by clinorotation. The second aim was to investigate morphological and gene expression changes in MLO-Y4 cells following suspension culture or clinorotation. We found that the ideal 3D scaffold consisted of 2% type I collagen mixed with 6% synthetic hydroxyapatite (collagen-HA). The scaffold was successfully formed into droplets for suspension culture, whereas it was adhered to the RCCS vessel walls for clinorotation. After 3 days of suspension culture, MLO-Y4 cells embedded in the collagen-HA droplets showed reduced expression of the mechanosensitive genes DMP1, E11, IL-6, and RANKL and the number of cells was elevated compared to the static control. We estimated that the drag force acting on the aggregate of droplets was in the range of 2.1 – 4.4 dynes/cm², speculated to provide a small but significant stimulatory effect on the cells located on the surface, which may de-sensitize them over the course of 3 days. Following clinorotation for 3 days, MLO-Y4 cells showed reduced expression of DMP1 and E11 genes while the cell number was unchanged compared to the static control. Since clinorotation and spaceflight cause the de-differentiation of other bone cells, we speculated that our MLO-Y4 cells were also de-differentiating during clinorotation, resulting in the lower expression of osteocyte-specific genes.