The Argentina international landed heavily five minutes into Sunday’s match after hurdling goalkeeper Salvatore Sirigu when trying to reach Miralem Pjanic’s throughball.
The 30-year-old tried to continue after receiving treatment but eventually had to be replaced by Federico Bernardeschi, with it appearing that Higuain had hurt his ankle.
It will come as a concern for Juve boss Massimiliano Allegri ahead of a crucial run of fixtures in his side’s season.
Juve take on Atalanta in Serie A next Sunday before the second leg of the Coppa Italia semi-final against the same opponents three days later.
They then begin March with a trip to Lazio, four days before they take on Tottenham in the Champions League last 16, who hold the edge in the tie on away goals after a 2-2 draw in Turin.
The loss of the Argentine, who scored both of his side’s goals against Spurs on Tuesday, was a big blow for the Serie A champions as they look to overtake Napoli at the top of the table.
However, Bernardeschi proved to be a sound replacement for the former Real Madrid and Napoli star as he took on a pass from Alex Sandro and drove into the box to set the Brazilian up with the return pass and give his side the lead after 33 minutes.
