The implantation of tissue-engineered constructs leads to hypoxic and physical stress to the seeded cells until they were reached by a functional microvascular system. Preconditioning of cells with heat shock induced heat shock proteins, which can support the cells to survive a subsequent episode of stress that would otherwise be lethal. Preconditioning of tissue-engineered constructs resulted in significantly higher number of surviving osteoblast-like cells (OLC). At the 6th and 10th day, angiogenic response was found comparative to poly(L-lactide-co-glycolide) (PLGA) scaffolds vitalized with either unconditioned or preconditioned OLC. However, they were significantly enhanced compared with the nonvitalized collagen-labeled PLGA scaffolds. This study demonstrates that vitalization of PLGA scaffolds with OLC accelerates the angiogenic response induced by the surrounding host tissue. In addition, heat shock preconditioning significantly enhances the survival rate of the OLC that are seeded on these scaffolds. Thus, vitalization of substitutes with adequately pretreated OLC may promise biologically adequate osseous restorations.