Display substrate and manufacturing method thereof, and display device

A manufacturing method of a display substrate, comprising forming an opening forming region, a display region at least partially surrounding the opening forming region, and an opening peripheral region between the opening forming region and the display region, wherein the manufacturing method further comprises:

• providing a base substrate,
• in the opening peripheral region, forming a first barrier dam on the base substrate and forming an insulating protection layer on a side of the first barrier dam away from the base substrate, wherein the first barrier dam at least partially surrounds the opening forming region, the insulating protection layer comprises at least one first material removal region, and a projection of the first material removal region on the base substrate is overlapped with a projection of the first barrier dam on the base substrate;
• and
• at least partially removing a material of the insulating protection layer in the at least one first material removal region, to form a recess portion in the insulating protection layer or a through hole penetrating the insulating protection layer.