Endothelial and Dental Pulp Cells Co-Culture for Bone Tissue Engineering

One of the biggest challenges in tissue engineering technique is the adequate vascularization as to prevent tissue necrosis. This problem often arises in large bone defect reconstruction. During osteogenesis, vascularization involves cross-talk regulation between endothelial and osteoblast cells and become the foundation of this study. Objective: To evaluate the effect of co-culture and conditioned medium of Human Umbilical Vein Endothelial Cells (HUVEC) on the expression of collagen I, fibronectin proteins and ALP activity released by dental pulp cells (DPC) on 3D scaffold composed of Hydoxyapatite/Tri Calcium Phosphate/Chitosan (HA/TCP/Chitosan). Methods: Three main research group were (1) DPC co-culture with various ratios of HUVEC (2) DPC cultured with various concentrations of HUVEC conditioned medium (3) DPC cultured with normal medium as the control. Results: HUVEC co-culture increased the protein expression of collagen I, Fibronectin and ALP activity. However the upregulation was higher when DPC were cultured with HUVEC conditioned medium. The increase in three different markers tested in conditioned medium compared to co-culture system might be explained by the presence of signaling molecules in the HUVEC conditioned medium that stimulate DPC. Fibronectin expression was necessary for DPC attachment to the 3D scaffold. While collagen I and ALP expression might indicated the differentiation of DPC to osteblasts. Conclusion: Conditioned medium HUVEC has a better potential to be used to stimulate the differentiation of dental pulp cells towards osteblastic lineage. The condition that is necessary for the success of bone tissue engineering. This study was financially supported by University of Indonesia Research Grant