I'm not clear if you asking about simply saving the project (PDS file), or if you're referring to packing the actual project and saving eerything, including all media clips, to a new folder.
PDS files don't store any media, and each time you open it PD will expect to find all the clips in their original folders. PD doesn't make any changes to the media at all, regardless of the additions or deletions to the project timeline and Media Room.
If you've packed a project then made changes to the media you've used, you'll need to pack the project again to retain copies of all the current clips. I highly recommend using a different folder and then deleting the original packed project folder to free up space on your hard drive.