The embodiment of the invention provides a display substrate and a preparation method thereof, and a display device. The display substrate comprises an underlayment substrate. Pixel defining layers are arranged on the underlayment substrate. Pixel areas used for containing ink drops are formed between the pixel defining layers. Each pixel area comprises a center area and an edge area surrounding the center area, and the surface energy of the edge area is larger than that of the center area. According to the embodiment of the invention, the risk of de-wetting of the display substrate can be reduced, and the printing quality can be improved.