Speaking at a ceremony marking the removal of the metal gates separating the Spanish town of La Línea de la Concepción from the tiny British territory, Sánchez said the end of routine border checks marked a historic moment.
"For decades, the border fence was exactly that, an open wound for the thousands of workers who crossed every day," the Socialist leader said.
"Today, we are making history, good history, because today, the last wall in continental Europe falls," he added, saying the agreement opened the doors to a "new era" of shared prosperity for the region.
The ceremony, attended by Gibraltar Chief Minister Fabian Picardo, took place after the end of border checks between Spain and the territory came into effect at midnight.
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The agreement between Britain and the European Union was signed on Tuesday in Brussels after years of wrangling following the UK's exit from the bloc in 2020.
A steady stream of pedestrians and vehicles crossed freely in both directions across the frontier on Wednesday. Some people held up their phones to record the moment.
Gibraltar is home to only about 40,000 people but relies on some 15,500 workers who cross from Spain every day, nearly half its workforce.
The border was closed by Spanish dictator Francisco Franco in 1969 after Gibraltar, which relies on London for defence and foreign policy, voted overwhelmingly in a referendum to remain British.
The closure, which lasted 13 years, cut off the daily movement of workers from Spain into Gibraltar and separated families.
Since then, long queues have repeatedly formed at the border when diplomatic tensions over the territory's sovereignty have led to tighter controls by Spain.
London and Madrid have disputed control of Gibraltar since the territory was ceded to Britain in the 1713 Treaty of Utrecht.
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