In virtue of the rapid metabolism and short half-life of adenosine (AD) in the human body, it is highly desired to explore new adenosine delivery system with controlled and steady release capacity. In this study, composite coatings consisting of polyethyleneimine (PEI), hyaluronic acid (HA) and collagen type I (COL-1) on titanium substrate surface are developed by layer-by-layer self-assembly technique, in which different concentrations of AD are encapsulated. In-vitro release study demonstrated a steady and sustained release of AD from the composite coating system. Adenosine within composite coatings depicted a concentration-dependent regulatory effect on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and osteoclastic differentiation of mouse leukemia cells of monocyte macrophage (RAW264.7) cells. All of these coatings exhibited good cell viability and promoted osteoblast cell proliferation. Among various composite coatings, the optimized coating specimen with loading of 65 μg AD (AD2) exhibits the best osteoconductive and osteoinductive activities with significantly enhanced osteoblast-associated gene expressions, and increased alkaline phosphatase (ALP) activity and calcium deposits. On the other hand, AD2 specimen shows the activity of inhibiting osteoclast differentiation and osteobclast-associated gene expressions as well as suppressing tartrate-resistant acid phosphatase (TRAP) activity. The superior performance of AD/PEI/HA/COL composite coatings indicated that this coating system could be useful for bone tissue engineering.