Dynamic cell culture on calcium phosphate microcarriers for bone tissue engineering applications

Abstract

Developing appropriate cell culturing techniques to populate scaffolds has become a great challenge in tissue engineering. This work describes the use of spinner flask dynamic cell cultures to populate hydroxyapatite microcarriers for bone tissue engineering. The microcarriers were obtained through the emulsion of a self-setting aqueous α-tricalcium phosphate slurry in oil. After setting, hydroxyapatite microcarriers were obtained. The incorporation of gelatin in the liquid phase of the α-tricalcium phosphate slurry allowed obtaining hybrid gelatin/hydroxyapatite-microcarriers. Initial cell attachment on the microcarriers was strongly influenced by the speed of the dynamic culture, achieving higher attachment at low speed (40 r/min) as compared to high speed (80 r/min). Under moderate culture speeds (40 r/min), the number of cells present in the culture as well as the number of microcarrier-containing cells considerably increased after 3 days, particularly in the gelatin-containing microcarriers. At longer culture times in dynamic culture, hydroxyapatite-containing microcarriers formed aggregates containing viable and extracellular matrix proteins, with a significantly higher number of cells compared to static cultures.

Silk/Fibroin Microcarriers for Mesenchymal Stem Cell Delivery: Optimization of Cell Seeding by the Design of Experiment

“…Our results indicated that hASCs were able to adhere to LFAMs in less than 2 h, under suitable conditions. In particular, the dynamic seeding of cells provided the best outcomes in comparison with static cultures, in terms of cell adhesion and viability, confirming previously published literature findings [64,65]. Moreover, the modality of the dynamic culture—intermittent or not—deeply influences final cell adhesion and subsequent expansion and differentiation.…”

Stimulation of Odontogenesis and Angiogenesis via Bioactive Nanocomposite Calcium Phosphate Cements Through Integrin and VEGF Signaling Pathways

“…4 They possess unique properties, including good osteoconductivity, biocompatibility, and injectability, and have found increasing applications in regenerative therapies. [4][5][6][7][8][9] We previously reported that calcium hydroxyapatite (HA)-based CPCs promoted growth and differentiation in human DPSCs. 5 However, CPCs are difficult to use alone as pulp-capping agents because they have limited antibacterial activity, long curing times, and low compressive strength.…”

From the macroscale to nanostructures: can tissue engineering recreate bone features?

“…Thus, turbulent forces and shear stress could be created, inducing cell sufferance. In such a bioreactor the Saos-2 osteoblastic cell line was shown to attach to HA-incorporating microcarriers and proliferate in a more efficient way than when the microcarriers were kept in static conditions [127]. Seeded into a decellularized bone-derived scaffold, adipose tissue-derived stem cells are able to adhere, migrate, grow and differentiate into osteoblastic cells expressing ALP.…”