The boys, aged 13 and 17, were both taken to hospital with stab wounds after the incident in Wednesfield Road, Heath Town, at around 2.15pm today.
One of the boys was seriously injured but West Midlands Police said neither of their injuries were believed to be life-threatening.
The A4124 was closed between Springfield Road and Woden Road while police and paramedics attended the scene.
The incident happened less than 24 hours after a man was shot in the leg in the city and came as the region’s police and crime commissioner warned that shootings and violence are on the rise as gangs "come out of lockdown".
There have been five gun attacks in the West Midlands this week.
A West Midlands Police spokesman said: "We were called to reports of disorder in Wednesfield Road, Wolverhampton, just after 2.20pm today (29 May).
"Two boys - aged 13 and 17 - have been taken to hospital with stab injuries which are not believed to be life-threatening.
"Enquiries are underway and anyone with information can contact us via Live Chat or by calling 101. Quote log number 2405 of 29/05/20."
A West Midlands Ambulance Service spokesman added: "We were called to reports of an assault on Wednesfield Road at 2.26pm and sent two ambulances, a paramedic officer and the Midlands Air Ambulance Critical Care Car to the scene.
"On arrival we discovered one male patient who was treated for serious injuries before being taken to hospital for further assessment."