Bioactive Glass Particles in Two-Dimensional and Three-Dimensional Osteogenic Cell Cultures

Abstract

This study aimed to investigate the influence of a three-dimensional cell culture model and bioactive glass (BG) particles on the expression of osteoblastic phenotypes in rat calvaria osteogenic cells culture. Cells were seeded on two-dimensional (2D) and threedimensional (3D) collagen with BG particles for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity was performed. Cell morphology and immunolabeling of noncollagenous bone matrix proteins were assessed by epifluorescence and confocal microscopy. The expressions of osteogenic markers were analyzed using RT-PCR. Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Experimental cultures produced a growing cell viability rate up to 14 days. Although ALP activity at 7 days was higher on BG cultures, cells on 3D and 3D+BG had an activity decrease of ALP at 14 days. Three-dimensional conditions favored the immunolabeling for OPN and BSP and the expression of ALP and COL I mRNAs. BG particles influenced positively the OC and OPN mRNAs expression and calcified nodule formation in vitro. The results indicated that the 3D cultures and BG particles contribute to the expression of osteoblastic phenotype and to differentiated and mineralized matrix formation.

Formation and Properties Polymer Nanolayers to Enhance Cell Growth in vitro

“…So it could be concluded that the stereochemistry of the polymers used for coating glasses greatly influence the cell growth and survival. This agrees with another study [13] and could be explained by the organization and maturation of the extracellular matrix surrounding cells by the created threedimensional system. An additional approach for the detection of cells viability is the determination of LDH levels.…”

Optimal Parameters of Laser Therapy to Improve Critical Calvarial Defects

“…The new bone formed into the cavity increased approximately 25%, 50%, and 55%, respectively, with 4, 7, and 11 sessions of LT and compared to referential Control values obtained in IBB-filled but not irradiated with lowlevel laser, i.e., combining the use of IBB with seven sessions of LT seems to be an optimal approach to greatly improve the recovery of calvarial defects. It would be important to verify if the optimal effects of seven sessions of LT on bone mineralization are also observed in cavities filled with other biomaterials such as the bioactive glass particles which have been used in in vitro studies from other investigators (Alves et al, 2017).…”

Bioengineering the ameloblastoma tumour to study its effect on bone nodule formation

“…Resident osteoblasts potentially clustered quicker in the stiff matrix than soft matrix, due to increased cellular and matrix density. We characterised this 3D bone stroma extensively, and did not limit the study to mineralisation assessments via ALP assay or alizarin red staining 34 . The alizarin red stain or von Kossa stain indicated mineralisation (dystrophic calcification) 65 .…”

“…We wanted to undertake extensive characterisation of 3D in vitro bone nodule formation in dense collagen matrices, to match, at a minimum, that done for 2D bone nodule formation, including mineralisation and gene markers. For confirmation of definitive in vitro bone nodule formation it was paramount to measure structure and composition as well as to investigate of specific gene panels for all stages of bone formation 34 – 36 .…”