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Tissue Engineering

Tissue Engineering - Poster Session E

Poster BB16 - Utilization of Forskolin Loaded Lipid Coated Mesoporous Silica Nanoparticles for Improved Angiogenesis in vitro and in vivo

Saturday, October 26, 2024
10:00 AM – 11:00 AM EST
Location: Exhibit Hall E, F & G

Introduction:  Obesity is a growing global health concern, often associated with metabolic disorders and cardiovascular diseases. One promising therapeutic strategy to combat obesity is the conversion of white adipose tissue to beige adipose tissue, a process that requires adequate vascularization. Forskolin (FSK), a natural compound known for activating adenylate cyclase and increasing cyclic AMP levels, has shown the potential to mediate this conversion. Recent advancements have led to the development of FSK-loaded lipid-coated mesoporous silica nanoparticles (LCMSN-FSK), which may enhance angiogenic properties. This study investigates the angiogenic properties of FSK and FSK-loaded nanoparticles both in vitro, using human umbilical vein endothelial cells (HUVECs), and in vivo. Understanding these properties could provide critical insights into how FSK contributes to the vascularization necessary for beige adipose tissue formation and its potential therapeutic applications in obesity treatment.

Materials and

Methods:  Mesoporous silica nanoparticles (MSNPs) were synthesized using a solution-based surfactant-directed self-assembly method conjugated with cy3-fluorescent dye. Liposomes were fused to MSNPs through electrostatic interactions to form the lipid-coated mesoporous silica nanoparticles (LCMSN). In vitro experiments involved plating HUVECs in a 24-well plate coated with a thin layer (250 µL) of growth factor-reduced (GFR) Matrigel. Five experimental groups were established: (G1) control (M199 media), (G2) single treatment of LCMSN-FSK (50 µg/mL M199 media), (G3) LCMSN-FSK treated at 6 and 12 hours, (G4) free FSK (5 µg/mL) only, and (G5) FSK only treated at 6 and 12 hours. Time-lapse imaging was conducted over 24 hours using a confocal microscope, with fluorescently labeled nanoparticles to monitor uptake and activity. In vivo studies were performed by injecting 12-week-old male C57BL/6J mice subcutaneously with LCMSN-FSK (1mg/mL dose), FSK (0.1mg/mL dose), and LCMSN (1mg/mL dose) as control. After 3 hours, the animals were sacrificed, and subcutaneous tissue was isolated. Angiogenesis gene markers, CD31 and VEGF, were quantified using RT-qPCR.

Results, Conclusions, and Discussions: Fluorescent images showed successful uptake of LCMSN over time. RT-qPCR results demonstrated significant elevation in CD31 expression when HUVEC were treated with two doses of LCMSN-FSK (p > 0.05) when compared to the other experimental groups. In vivo, expression of CD31 and VEGF was also significantly increased (p>0.05 and p>0.01) when compared to free FSK and LCMSN treatment groups, indicating an increase in angiogenesis factors. Thus, LCMSN-FSK has demonstrated effectiveness in enhancing pro-angiogenesis RNA expression

Acknowledgements (Optional):