Twelve people, including some of France's most celebrated cartoonists, were killed on January 7, 2015, when brothers Said and Cherif Kouachi went on a gun rampage at the paper's offices in Paris.
In the centre of the cover is a cartoon of the prophet drawn by cartoonist Jean Cabut, known as Cabu, who lost his life in the massacre.
"All of this, just for that," the front-page headline says.
The latest Charlie Hebdo cover shows a dozen cartoons first published by the Danish daily Jyllands-Posten in 2005 -- and then reprinted by the French weekly in 2006, unleashing a storm of anger across the Muslim world.
The editorial team wrote that now was the right time to republish the cartoons and "essential" as the trial opens.
"We will never lie down. We will never give up," director Laurent "Riss" Sourisseau wrote in an editorial to go with the cartoons in the latest edition.
"The hatred that struck us is still there and, since 2015, it has taken the time to mutate, to change its appearance, to go unnoticed and to quietly continue its ruthless crusade," he said.
"We have often been asked since January 2015 to print other caricatures of Mohammed," it said.
"We have always refused to do so, not because it is prohibited -- the law allows us to do so -- but because there was a need for a good reason to do it, a reason which has meaning and which brings something to the debate."
The paper's willingness to cause offence over a range of controversial issues has made it a champion of free speech for many in France, while others argue it has crossed a line too often.
"The freedom to caricature is guaranteed for all, the freedom to love or not to love (the caricatures) as well. Nothing can justify violence," he told AFP.