RESUMEN
Cellulose nanocrystals (CNCs) are sustainable particles that are effective at stabilizing emulsions by adsorbing at droplet interfaces and providing a steric barrier to coalescence. However, CNCs have surface charges that reduce the coverage of the emulsion droplets due to the electrostatic repulsion between CNCs. In such cases, adding salt is a typical (and straightforward) way to adjust the formulation so that the charges are screened, allowing increased coverage of the droplets. At the outset of this work, we hypothesized that characterization of the interfacial tension and interfacial shear rheology of the oil-water interface would be correlated to interfacial coverage and therefore predictive of the optimal salt concentration for emulsion stability. Included in the methods section as a useful reference to others is the presentation of a detailed derivation for the equations needed to compute interfacial shear moduli in a custom, double-gap geometry. In contrast to our hypothesis, we found that interfacial tension did not correlate well with emulsion stability and that the native surface-active compounds in corn oil overwhelmed any influence of the CNCs on the interfacial tension. Additionally, we found that interfacial shear rheology (which can be painstakingly difficult to measure) was not a useful tool for formulating these emulsions. This is because at commonly used concentrations of CNCs, the bulk rheology is increased to a much greater degree than that of the interface, making the details of the interfacial rheology unimportant. Finally, we found that at concentrations of CNCs that are typical in industrial processes, characterizing the bulk viscoelastic properties of the aqueous suspending phase without added oil (a relatively simple measurement) is sufficient to predict the influence of NaCl concentration on charge screening between the CNCs and, by extension, increased surface coverage of droplets for greater emulsion stability.
RESUMEN
Purpose: The rotary cell culture system (RCCS) is a common clinorotation device for cell culture. It is also used as a low-shear suspension culture bioreactor to form functionalized 3D tissue constructs and to model microgravity. We sought to develop a 3D scaffold composed of type I collagen and hydroxyapatite (collagen-HA) to characterize MLO-Y4 osteocytes following suspension culture or clinorotation.Materials and Methods: MLO-Y4 cells were embedded in collagen-HA. The scaffold was formed into droplets for suspension culture or wall-adhered to the RCCS for clinorotation. AFM, rheometry, immunofluorescence and qRT-PCR were employed to measure the scaffold stiffness, cell viability and gene expression of cells in collagen-HA scaffolds. Dendritic cells were visualized and quantified and gene expression after suspension culture and clinorotation was compared to static controls.Results: The optimized scaffold for the RCCS consisted of collagen with 6 mg/mL HA which had a stiffness of < 1 kPa. MLO-Y4 cell viability was higher in collagen-HA scaffolds, compared to scaffolds without HA. Collagen-HA scaffolds induced higher osteocyte-specific gene expression compared to cells cultured on 2D plastic. Cells in the scaffold downregulated DMP1, E11, IL-6, and RANKL, and had fewer dendritic cells following suspension culture whereas clinorotation downregulated DMP1 and E11 genes, compared to static controls.Conclusions: Suspension culture for 3 days in collagen-HA stimulates growth of osteocytes but may also desensitize them to mechanical cues. Clinorotation for 3 days in collagen-HA does not stimulate proliferation or expression of mechanosensitive genes, indicating that it may be an effective mechanical unloading environment.
