In theory, no, but if you're using an AMD Ryzen CPU, there's some nasty bugs related to VME instructions (which AMD appears to have no intention of fixing) that break some DOS-based applications, including Windows 3, 9x, and probably the other DOS-based Windows versions as well but I never tried them. NT-based Windows works, though early versions do not perform as well as they do on Intel (my R7 1700 and 5700X run Windows 2000 VMs worse than my old i5-6600...I wish I was joking).
