Halloysite nanotube-embedded microparticles for intestine-targeted co-delivery of biopharmaceuticals.

Microparticles (MPs) with pH-responding macropores have recently proved their significance for the delivery of vulnerable biomolecules for oral drug administration. The previous MP systems were proven to provide enhanced protection against the gastric environment, however, their application is hindered due to insufficient loading efficiencies and deficient penetration capabilities of encapsulated drugs across the mucus barrier. Here, we report a new co-delivery approach based on amine-functionalized halloysite nanotube (HNT)-embedded MPs (amine-HNT-MPs) with pH-responding macropores specifically designed to deal with the mucus barrier at the absorption site. The mean diameter and polydispersity index of the pored MPs were measured by a particle size analyzer to be 37.6 ± 1.3 µm and 1.15, respectively. The drug loading capacity of the co-delivery system was shown to be 50-times higher than previously reported pored MPs. Fluorescence microscopy analysis of sulforhodamine B (into a hollow interior of HNTs)/ fluorescent nanoparticles (into a hollow interior of MPs)-encapsulated MPs confirmed biphasic release behavior due to pH-dependent pore closing/opening in the simulated gastrointestinal (GI) digestive conditions. To verify the protective effect of the co-delivery system, bromelain and lactase were loaded into HNTs and MPs, respectively, and found to exhibit 94.5 ± 3.3% (bromelain) and 70 ± 14.1% (lactase) functional activity in simulated GI tract conditions. The considerable improvement in the stability of the encapsulated enzymes against gastric conditions are attributed to the efficient pore sealing of the co-delivery system after the encapsulation of enzymes and maintenance of these closed pores in the gastric environment. Furthermore, the mucolytic enzyme (i.e. bromelain)-encapsulated co-delivery system was found to enhance mucopenetration of the encapsulated drug from histological analysis using ex vivo porcine intestine tissue. Therefore, the new microencapsulation design proposed in this study provides a promising solution to the major issues hampering the wide-spread application of MPs in the development of oral drug formulations for biopharmaceuticals and vaccines.

Metabolic properties of astrocytes differentiated from rat neurospheres.

The metabolic properties of astroglia differentiated from neurospheres have not been fully assessed. In this study, the glycolytic and oxidative metabolism of glucose in astroglia differentiated from rat tertiary neurospheres (astroglia(NS)) was compared with that in astroglia prepared from the striata of embryonic day 16 rats (astroglia(ST)). In addition to the basal condition, we also investigated energy metabolism under Na+,K+-ATPase activation. Furthermore, the effects of glucose concentration in the culture medium were assessed. No significant differences in 2-deoxy-D-[1-(14)C]glucose phosphorylation were observed between astroglia(NS) and astrogliaST. The rates of L-[U-14C]lactate ([14C]lactate) and D-[U-14C]glucose ([14C]glucose) oxidation were 5.74+/-0.82 and 2.83+/-0.4 pmol/60 min/microg protein, respectively, in astrogliaNS grown in low glucose (2 mM) and 3.01+/-1.03 and 1.77+/-0.23 pmol/60 min/microg protein, respectively, in astrogliaNS grown in high glucose (22 mM). Neither the [14C]lactate nor the [14C]glucose oxidation rates in astrogliaNS were significantly different from those in astrogliaST. D-aspartate (500 microM) significantly increased the [14C]lactate and [14C]glucose oxidation rates by 127% and 62%, respectively, in astrogliaNS grown in low glucose and by 217% and 115%, respectively, in astroglia(NS) grown in high glucose. D-aspartate also increased the oxidation of [14C]lactate and [14C]glucose to 236% and 147% of the control values, respectively, in astrogliaST grown in low glucose and to 174% and 144%, respectively, in astrogliaST grown in high glucose. Rat astroglia differentiated from neurospheres might possess an equivalent capacity for utilizing energy substrates under both basal and activated conditions to that of astroglia prepared from striatum.