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The number of planes that could take off and land at Gatwick Airport was restricted because of "short notice sickness" in the air control tower, managed by National Air Traffic Services (NATS).
EasyJet - the worst-affected airline with 42 cancellations - said staffing problems had caused 29 days of travel restrictions at Gatwick since May.
Its chief executive, Johan Lundgren, said customers were "being let down once again" and is calling for a "wide-ranging review" of NATS, which has been approached for comment.
A reduced schedule is being "imposed on airlines" because of the staffing situation, EasyJet added.
Among the other carriers forced to cancel flights on Wednesday were Lufthansa, Norwegian, Iberia Express and Vueling.
Passengers are not entitled to compensation because the disruption was out of the airlines' control.
This latest disruption follows the "one in 15 million" technical glitch which caused widespread difficulties last week.
NATS said those problems, at one of the busiest times of the year for travel on the August bank holiday, were caused by flight data which its system "didn't understand" and "couldn't interpret".
A Gatwick Airport spokeswoman said: "Due to short notice sickness in the air traffic control tower, temporary air traffic control restrictions were put in place (on Wednesday night).
"London Gatwick would like to apologise to any passengers who have been impacted by these restrictions. Please contact your airline for more information."
Read more:
'Unreliable' data caused air traffic error
How stranded passengers were delayed for hours
The Civil Aviation Authority (CAA) announced on Wednesday that an independent review will be carried out into last week's air traffic control failure.
Passengers departing from UK airports and ports are also being subjected to longer waiting times because of security checks during the search for an escaped prisoner.
Daniel Abed Khalife, 21, a former soldier accused of terrorism, went missing in his cook's uniform from HMP Wandsworth in southwest London on Wednesday.