Full-back David Bortolussi and winger Kaine Robertson are out of Italy's next Six Nations match against Wales in Cardiff on Saturday, February 23.
Bortolussi has a shoulder injury and Robertson a thigh strain, both sustained in Italy's 19-23 loss to England in Rome on Sunday.
Robertson will be out for a couple of weeks, doctors said.
Further tests are needed to determine the extent of Bortolussi's injury.
Paolo Buso, winning his first cap, has been called up to replace Bortolussi while Ludovico Nitoglia is in for Robertson.
Buso and Nitoglia are clubmates at Cammi Calvisano.
Italy are bottom of the Six Nations table along with Scotland but coach Nick Mallet is optimistic they can salvage something from the tournament.
A week before the four-point home loss to England they went down by five points, 16-11, to Ireland in Dublin.
On both occasions, and especially against England, Italy appeared to finish more strongly than their opponents.
After their trip to Cardiff Italy play group leaders France in Paris on March 9 and end their campaign at home to Scotland on March 15.
