Droplet microfluidics as a tool for production of bioactive calcium phosphate microparticles with controllable physicochemical properties

Abstract

Affordable and therapeutically effective biomaterials are required for successful treatment of orthopaedic critical-size bone defects. Calcium phosphate (CaP) ceramics are widely used for bone repair and regeneration, however, further optimization of their properties and biological performance is still required. To improve the existing CaP bone graft substitutes, novel synthesis and production approaches are needed that provide a fine control over the chemical and physical properties and versatility in the delivery format. In this study, a microfluidic strategy for production of CaP microparticles with different sizes derived from highly monodisperse droplets is proposed for the controlled synthesis of bioactive CaP ceramics. Microfluidic droplets, that served as microreactors for CaP precipitation, allowed the production of different CaP phases, as well as strontium-substituted CaP. By varying the concentration of the precursor solution, microparticles with different porosity were obtained. The droplet microfluidic system allowed direct visualization and quantification of the reaction kinetics. Upon production and purification of the microparticles, the biocompatibility and bioactivity were tested in vitro using human mesenchymal stromal cells (hMSCs). Cell attachment was analysed by imaging of the cytoskeleton and focal adhesions Moreover, cell proliferation, metabolic activity, alkaline phosphatase activity and mRNA expression of a set of osteogenic markers were quantified. We demonstrated that droplet microfluidics is a functional technique for the synthesis of a range of bioactive CaP-based ceramics with controlled properties. STATEMENT OF SIGNIFICANCE: Calcium phosphate (CaP) ceramics are widely applied synthetic biomaterials for repair and regeneration of damaged bone; yet, CaP bone graft substitutes require further improvement to fully replace natural bone grafts in challenging clinical situations. To this end, novel synthesis and production approaches are needed that provide a fine control over the chemical and physical properties. Here, we developed a microfluidic platform for production of CaP microparticles with different size, composition and porosity, derived from monodisperse droplets. We demonstrated that CaP microparticles produced using this platform supported growth and differentiation of human mesenchymal stromal cells. This platform is a useful tool for developing a variety of CaPs in a controlled manner to study their physicochemical properties in relation to their bioactivity.

Keywords: Bone graft substitutes; Calcium phosphate ceramics; Droplet microfluidics; Mesenchymal stromal cells.