Bambino Gesu Hospital had offered to treat the 11-month-old boy while his parents were trying to persuade UK courts to tell Great Ormond Street Hospital in London to release the boy for treatment in the United States.
Hospital chiefs said in a statement that they are not in a position to know what might have happened six months ago and cannot know if Charlie would have responded to the experimental therapy.
Hospital director Mariella Enoc told reporters that they carried out a clinical evaluation and offered medical assistance free of charge.
Meanwhile, Charlie Gard's parents are embroiled in a new High Court fight with doctors over the circumstances of the terminally-ill baby's death.
The little boy's mother Connie Yates was back in court on Tuesday a day after she and partner Chris Gard abandoned legal action over treatment for the baby.
A lawyer representing the couple told a judge that the couple wanted to take Charlie home to die.
But lawyers representing the hospital said there were practical problems and the couple had put forward no clear plan.
Mr Justice Francis was told that the hospital had suggested a "hospice option".
He said the dispute cried out for settlement.
Barrister Grant Armstrong, who leads the couple's legal team, suggested to Mr Justice Francis that hospital bosses were placing obstacles in Charlie's parents' way.
But barrister Katie Gollop QC, who leads Great Ormond Street's legal team, said practicalities were of the "greatest importance" but Charlie's parents had proposed no clear plan.
Ms Gollop said bosses wanted to fulfil Charlie's parents' "last desire".
But she said medics wanted to avoid hazards or mishaps and wanted to ensure Charlie was safe.
