One-day Treatment of Small Molecule 8-Bromo-cyclic AMP Analogue Induces Cell-based VEGF production for In Vitro Angiogenesis and Osteoblastic Differentiation
Life Sciences | Medicine and Health Sciences
Small molecule based regenerative engineering is emerging as a promising strategy for regenerating bone tissue. Small molecule cAMP analogues have been proposed as novel biofactors for bone repair and regeneration, and while promising, the effect that these small molecules have on angiogenesis, a critical requirement for successful bone regeneration, is still unclear. Our previous research demonstrated that the small molecule cAMP analogue 8-bromoadenosine-3’,5’-cyclic monophosphate (8-Br-cAMP) was able to promote initial osteoblast adhesion on a polymeric scaffold via cAMP signaling cascades. Here, we report that 8-Br-cAMP is capable of inducing in vitro cell-based VEGF production for angiogenesis promotion. We first demonstrated that treating osteoblast-like MC3T3-E1 cells with 8-Br-cAMP for one day significantly increased VEGF production and secretion. We then demonstrated that 8-Br-cAMP induced cell-secreted VEGF is biologically active and may promote angiogenesis as evidenced by increased endothelial cells (HUVECs) migration and tubule formation. In addition, treatment of MC3T3-E1 cells with 8-Br-cAMP for as short as a single day resulted in enhanced ALP activity as well as matrix mineralization, demonstrating in vitro osteoblastic differentiation. A short term 8-Br-cAMP treatment also addresses the concern of non-specific cytotoxicity, as our data indicate that a one-day 8-Br-cAMP treatment scheme supports cellular proliferation of MC3T3-E1 cells as well as HUVECs. While the major concern associated with small molecule drugs is the risk of non-specific cytotoxicity, the short exposure treatment outlined in this paper provides a very promising strategy to mitigate the risk associated with small molecules.
Lo, Kevin W.-H.; Kan, Ho Man; Gagnon, Keith A.; and Laurencin, Cato T., "One-day Treatment of Small Molecule 8-Bromo-cyclic AMP Analogue Induces Cell-based VEGF production for In Vitro Angiogenesis and Osteoblastic Differentiation" (2013). UCHC Articles - Research. 276.
J Tissue Eng Regen Med. Author manuscript; available in PMC 2015 Jul 9. Published before final editing as: J Tissue Eng Regen Med. : 10.1002/term.1839. Published online 2013 Nov 6. doi: 10.1002/term.1839 PMCID: PMC4497951 NIHMSID: NIHMS592371