In this report, a novel nanocomposite based on type I collagen (Col) was prepared, which contained varying amounts (15.1, 30.2, and 75.5 ppm) of silver nanoparticles (AgNPs). The surface morphology and chemical composition pure Col and Col-AgNP nanocomposites (Col-Ag) was characterized by UV–Vis spectroscopy (UV–Vis), atomic force microscope (AFM) and Fourier transform IR spectrometer (FTIR). The biocompatibility effect and biological activity of Col-Ag culturing with mesenchymal stem cells (MSCs), as well as guiding for angiogenesis differentiation, were evaluated via in vitro assay. The Col-Ag in 30.2 ppm demonstrated better biological properties and compatibility culturing with MSCs. The biological properties could be associated with cell adhesion, proliferation, migration and differentiation. Afterwards, the induced angiogenesis and differentiation of MSCs by the expression of von Willebrand Factor (vWF) and CD31 were also investigated. Furthermore, both anti-inflammatory and endothelialization ability were also investigated in vivo assay. It was observed that Col-Ag nanocomposites not only inhibited CD86 expression, but also facilitated endothelialization capacity, the expression of CD31 when implanting Col-Ag into rats subcutaneously after 4 weeks. This current research indicates that Col-Ag nanocomposites has potential of being employed as a surface modification approach, and is better in clinical treatments with MSCs for vascular regeneration applications.