Dynamic cultivation with radial flow bioreactor enhances proliferation or differentiation of rat bone marrow cells by fibroblast growth factor or osteogenic differentiation factor

Abstract

Dynamic cultivation using a radial flow bioreactor (RFB) has gained increasing interest as a method of achieving bone regeneration. In order to enhance bone generation in large bone defects, it is necessary to use an RFB to expand the primary cells such as bone marrow cells derived from biotissue. The present study aimed to evaluate the cell expansion and osteogenic differentiation of rat bone marrow cells (rBMC) when added to basic fibroblast growth factor containing medium (bFGFM) or osteogenic differentiation factor containing medium (ODM) under dynamic cultivation using an RFB. Cell proliferation was evaluated with a DNA-based cell count method and histological analysis. An alkaline phosphatase (ALP) activity assay and immunohistochemistry staining of osteogenic markers including BMP-2 and osteopontin were used to assess osteogenic differentiation ability. After culture for one week, rBMC cell numbers increased significantly under dynamic cultivation compared with that under static cultivation in all culture media. For different culture media in dynamic cultivation, bFGFM had the highest increase in cell numbers. ALP activity was facilitated by dynamic cultivation with ODM. Furthermore, both BMP-2 and osteopontin were detected in the dynamic cultivation with ODM. These results suggested that bFGFM promotes cell proliferation and ODM promotes osteogenic differentiation of rBMC under dynamic cultivation using an RFB.

The roles of signaling pathways in bone repair and regeneration

“…Basic FGF (bFGF), is a potent mitogen for MSCs (Martin, Muraglia, Campanile, Cancedda, & Quarto, ), which resulted in incorporation of a higher number of osteocytes into healing bone when treated with bFGF (Kawaguchi et al, ). Kanda et al (). reported that culture of the rat bone marrow cells (rBMC) in bFGF containing medium resulted in a significant increase in cell number and cell expansion, as well as expression of the BMP‐2 and osteopontin.…”

Nanolamellar Tantalum Interfaces in the Osteoblast Adhesion

“…Alkaline phosphatase (ALP), as an early marker for osteogenic differentiation, was used as an indicator of osteoblastic mineralization . As shown in Figure a, the ALP activities of the osteoblasts were evaluated on days 7 and 14 on Ta­(B), Ta40, and Ta70 cultured with MC cells.…”

“…Alkaline phosphatase (ALP), as an early marker for osteogenic differentiation, was used as an indicator for osteoblastic mineralization. 53 As shown in Figure 7(a), the ALP activity of the osteoblasts were evaluated at days 7 and 14 on Ta(B), Ta40 and Ta70 cultured with MC cells. The ALP activity on bulk Ta(B) at day 7, was lower than those of two nanolamellar Ta surfaces, and the activity on Ta70 was the highest.…”

Perfused Platforms to Mimic Bone Microenvironment at the Macro/Milli/Microscale: Pros and Cons

“…Interstitial fluid flow modulates osteogenesis and can be recapitulated in vitro at the milliscale level through the generation of fluid shear stress or through the application of mechanical strains ( Alfieri et al, 2019 ; Ramani-Mohan et al, 2018 ), by using both commercially available and custom-made perfusion bioreactors ( Tseng et al, 2014 ; Kleinhans et al, 2015 ; Kanda et al, 2016 ; Beşkardeş et al, 2018 ; Burgio et al, 2018 ; Ramani-Mohan et al, 2018 ; De Luca et al, 2020 ). U-Cup bioreactors are the most widely used among those offered by the market, as they effectively perfuse media into the scaffold through a pump system.…”