Syrian regime air strikes hit the Sakhour hospital and a health center twice last week in an opposition-held district of the northern city of Aleppo.
The hospital suspended all of its operations after damage caused by missiles and barrels bombs rendered the facility unusable. Red crescent health-care workers were working in the facility.
According to Doctors Without Borders least 40,000 people had been treated at this hospital.
Last month Doctors Without Borders said that more than 300 trauma surgeries were performed in the hospital.
Health aid workers affirmed that the hospital has completely stopped functioning and was fully evacuated.
The same hospital was closed as well in 2014 for weeks following damages from air strikes.
"It is unclear if or when the hospital will be operative again," Doctors Without Borders, known by its French acronym MSF, said in a statement.
"These new attacks on medical infrastructures are intolerable," said Raquel Ayora, MSF's director of operations.
Another health center was closed as well after having been targeted earlier this month, the group said.
Later on Sunday a warplane also attacked a school in Aleppo, killing seven people including children.
According to the Syrian Observatory for Human rights, toxic gases were also used Monday for the eighth time in the Idlib province, which opposition forces took over last month.
Amnesty International said on Tuesday that the Syrian regime is committing "crimes against humanity" by conducting air strikes against civilians in Aleppo, and also criticizing opposition fighters for what they called "war crimes" being committed on “a daily basis”.
Philip Luther, Amnesty's Middle East and North Africa program director, said "By relentlessly and deliberately targeting civilians the Syrian government appears to have adopted a callous policy of collective punishment against the civilian population of Aleppo,"
Last year, at least 3,000 civilians were killed in Aleppo Province by Syrian regime barrel bomb attacks, according to Amnesty.
After the war flared up in Aleppo, the commercial capital of Syria, in 2012, the city was divided into two parts with Assad regime forces continually losing ground to opposition groups.
More than 220,000 people have been killed in Syria since March 2011, and more than 1 million have been wounded since the armed conflict started in 2012, according to United Nations estimates.