A display substrate and a manufacture method thereof, and a display device are provided. The display substrate includes a display region, the display region includes an organic functional layer, an encapsulation layer, and a first barrier wall; the display region has a first opening, the first barrier wall surrounds an outer edge of the first opening, the organic functional layer surrounds the first barrier wall, and the encapsulation layer covers the organic functional layer and the first barrier wall; the encapsulation layer includes a first portion and a second portion, the first portion is configured to cover a portion of the organic functional layer close to the first opening; a second portion is configured to cover a side of the first barrier wall adjacent to the organic functional layer, the first portion forms an obtuse angle with the second portion.