Hotfixes are program updates. Conceivably they could change the way files are written to or maintained and could change the MDB's indirectly, but I would not expect a structure change from a hotfix. Most likely, if anything, queries would execute differently.
The database engine runs on each client. So each user that is experiencing the problem would need the update. Although you should be able to test with no one else using the database before and after the update to see if the hotfix solves your problem (from other posts I know you are referencing a performance hotfix). Then you can test it for a while before giving it to everyone else for issues.
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