Enhanced antioxidant and anti-inflammatory effects of bee pollen and honey hybrid formulation by hot-melt extrusion

Modifying the complex wall structure of bee pollen is a way to enhance the uptake of bioactive compounds. Bee Pollen and Bee Honey hybrid formulation (BPHF) are made by hot melt extrusion (HME), which is known to be the best cell wall structure-breaking method for active compound extraction because it can break the fibrous matrix of the compounds. Honey and bee pollen mixtures showed increased antioxidant activity with increased phenolic and flavonoid contents and decreased inhibitory concentration 50 (IC₅₀) values compared to the control group. In addition, the content of gallic acid, one of the phenolic acids, increased about four times in the HME-F3 formulation compared to the control group, indicating that the phenol content was rich, but the HME-F3 formulation decreased in comparison. All samples had nano-sized particles. The Polydispersity index (PDI) value of the HME-F3 formulation was reduced and well distributed, and the stability of the particles was confirmed in the zeta potential results. Scanning electron microscopy (SEM) showed the image change of the sample after HME compared to the existing bee pollen image, and Transmission electron microscopy (TEM) showed that the particles of the mixed formulation were uniformly dispersed. Fourier-transform Infrared Analysis (FT-IR) results showed that there were structural changes in the mixture formulation compared to the control, and X-ray Diffraction Analysis (XRD) results showed that all samples were amorphous. It was found that the HME-F3 formulation could resist inflammation and protect the function of the intestinal epithelial wall. The mRNA levels of TJ-related proteins in the mixed formulation were reduced compared to LPS treatment and control.