Polytetrafluoroethylene (PTFE) is a kind of fluoropolymer with many critical applications. Nonionic surfactant stabilized concentrated PTFE aqueous dispersions are widely used to prepare corresponding products via dipping, spraying, spinning, or film casting, as these surfactants exert a negligible effect on the properties of the products. Although many nonionic surfactants have been used to stabilize PTFE dispersions in industry, the effects of molecular structure and topology are not well-documented; thus, currently, there is no basis for selecting or designing suitable novel surfactants to enhance dispersion stability. In this work, five nonionic surfactants with different topological structures including Tween 20, Tween 60, Tween 80, Brij L23, and Brij O20, were used to prepare concentrated PTFE dispersions. The variation of particle morphology before and after concentration was examined by scanning electron microscopy, and the static stability of the dispersions was quantitatively and objectively characterized using a Turbiscan analyzer based on multiple light scattering. Then the effect of surfactant molecular structure and topology on dispersion stability was discussed. We found that, after the enrichment process, the PTFE microspheres were tightly packed, and some were deformed. Brij surfactants which bears a linear hydrophilic oxyethylene (EO) chain can form a thicker hydration layer on the surface of PTFE particles than those of Tween surfactants which contains a branched hydrophilic EO chain. Accordingly, PTFE dispersions with Brij surfactants show better static stability. Moreover, in comparison with the saturated hydrophobic tail, the unsaturated hydrophobic chain is more flexible, so it can adhere to the surface of PTFE particles better, improving the stability. In addition, the increase of surfactant concentration contributes to the increase of micelles density and viscosity of the bulk dispersion, which can also enhance the stability. These findings are beneficial to design or choose suitable surfactants for high stability and high-quality concentrated PTFE dispersions.