Harnessing adipose-derived stem cells to release specialized secretome for the treatment of hepatitis B

Mesenchymal stem cells (MSCs) have the function of repairing damaged tissue, which is known to be mediated by the secretome, the collection of secretory materials shed from MSCs. Adjusting the culture conditions of MSCs can lead to a significant difference in the composition of the secretome. It was hypothesized that pre-sensitization of MSCs with specific disease-causing agents could harness MSCs to release the therapeutic materials specialized for the disease. To validate this hypothesis, the present study aimed to generate a 'disease-specific secretome' for hepatitis caused by hepatitis B virus using hepatitis BX antigen (HBx) as a disease-causing material. Secretary materials (HBx-IS) were collected following the stimulation of adipose-derived stem cells (ASCs) with 100-fold diluted culture media of AML12 hepatocytes that had been transfected with pcDNA-HBx for 24 h. An animal model of hepatitis B was generated by injecting HBx into mice, and the mice were subsequently intravenously administered a control secretome (CS) or HBx-IS. Compared with the CS injection, the HBx-IS injection significantly reduced the serum levels of interleukin-6 and tumor necrosis factor-α (pro-inflammatory cytokines). Western blot analysis and immunohistochemistry of the liver specimens revealed that the HBx-IS injection led to a higher expression of liver regeneration-related markers, including hepatocyte growth factor and proliferating cell nuclear antigen, a lower expression of pro-apoptotic markers, such as cleaved caspase 3 and Bim in mouse livers, and a lower expression of pro-inflammatory markers (F4/80 and CD 68) compared to the CS injection. HBx-IS exhibited higher liver regenerative, anti-inflammatory and anti-apoptotic properties, particularly in the mouse model of hepatitis B compared to CS. This suggests that the secretome obtained by stimulating ASCs with disease-causing agents may have a more prominent therapeutic effect on the specific disease than the naïve secretome.