The surprising 6-1 6-4 loss that a lacklustre Osaka suffered on Tuesday could be explained away by the fact the 23-year-old had not played any competitive tennis since pulling out of the French Open at the end of May.
All the same, it was a major upset as world number 42 Vondrousova took out the highest remaining seed in the draw – the Japanese star who lit the Olympic cauldron on Friday.
Osaka's exit, after previous shock defeats for top seed Ash Barty and number three Aryna Sabalenka, has raised the prospect of a shock champion, just as occurred five years ago at the Rio Games when Monica Puig of Puerto Rico caused a sensation.
Now at the quarter-final stage, there is one former grand slam champion left in the field and two finalists at that level, but it really looks like anyone's title.
VONDROUSOVA SENSES AN OPPORTUNITY
It was remarkably straightforward for Vondrousova at Ariake Tennis Park, as she cruised through the opening set and soon reeled in Osaka's early break in the second.
Osaka saved two match points when serving to stay in the contest, but not a third, planting a backhand wide.
Considering Vondrousova reached the French Open final two years ago, in front of packed grandstands rather than the empty seats in Tokyo, it was no surprise she hesitated when asked whether this win over Osaka was the biggest of her career. It probably doesn't have that cachet, good a win though it was.
"Of course it's one of the biggest," Vondrousova said.
"Naomi is a great player, she has so many grand slams, so I knew it would be a tough match. But I'm just very happy with my play. I played amazingly in the first set, and then the second set was really tough. I'm just happy to be through.
"I think she was struggling a bit with my serving. Also, I use drop-shots very well. I'm just very happy with my game today."
She faces Spain Paula Badosa next and said: "It's very open now. I think every girl is playing really well. Now it's the quarter-final, so we'll see."
HAS SVITOLINA'S TIME ARRIVED?
A fixture in the top 10 over recent seasons, Svitolina has been unable to transfer her regular tour form onto the major stage on a consistent basis.
Maybe the Olympics will be a platform towards success on that stage, with Svitolina now the highest seed remaining in the draw, at number four. The Ukrainian is also on a high on the personal front, having married French tennis star Gael Monfils shortly before heading to Tokyo.
Two semi-finals, at Wimbledon and the US Open in 2019, have been her deepest runs in the majors, and this season has been one of diminishing returns, with a fourth-round run in Australia followed by a third-round Roland Garros exit and a round-two loss at Wimbledon.
Svitolina beat Maria Sakkari of Greece 5-7 6-3 6-4 on Tuesday, setting up a quarter-final against Italian Camila Giorgi who won 6-4 6-2 against Wimbledon runner-up Karolina Pliskova.
"I don't think I'm a favourite because there are lots of good players here and everyone is quite equal," Svitolina said.
A MUG SHOT?
Should Spain's Garbine Muguruza be considered the favourite from this point? With French Open and Wimbledon titles in her trophy room, Muguruza has shown she has what it takes to triumph on a big stage, and a clinical 6-4 6-1 win over Belgian Alison Van Uytvanck on Tuesday was just the job.
She goes on to face Elena Rybakina of Kazakhstan, who edged past Croatian Donna Vekic.
Belinda Bencic of Switzerland caused a surprise by ousting the in-form reigning French Open champion Barbora Krejcikova, springing a 1-6 6-2 6-3 win that means there will be no repeat of the Roland Garros final in the quarter-finals.
That had been on the cards, but Bencic will be the player who takes on Anastasia Pavlyuchenkova for a place in the final four.
Pavlyuchenkova scored an impressive 6-1 6-3 victory over Sara Sorribes Tormo of Spain, the player who knocked out Barty in round one.
Russian Olympic Committee's Pavyluchenkova is looking to harness the form that took her to a maiden slam final, describing her Paris run as "a great experience to have".
"But every week is a new week and this is a new event," said the 30-year-old. "The Olympic Games is a very special event. It's different. It's nothing like the others."