Messi and Inter Miami eliminated from MLS play-off contention
DC United also missed out on a spot in the post-season and immediately announced that their English manager, former Manchester United star Wayne Rooney, was leaving his post by "mutual agreement".
Messi returned from injury as a substitute in the 55th minute but Cincinnati, who have earned top seed in the play-offs, grabbed the win with a goal from Argentine Alvaro Barreal.
Messi made little impact on the field and although he had two free-kick opportunities within his shooting range, both his efforts sailed harmlessly wide.
"I could see that he was lacking football (game time)," said Miami head coach Gerardo Martino.
"The injury is fine, he doesn't have any problems, but (his performance was) logical because in recent times he has played very little and it could happen that he is lacking in rhythm, which is also why he only played 32 or 33 minutes," he added.
With other results going against Miami, Messi's side were left next to bottom of the Eastern Conference, seven points off the last play-off spot with just two games remaining.
Messi had played just 37 minutes for his club since September 3rd, having returned with an injury from international duty with Argentina last month.
Without him Miami struggled, paying the price for their successful run to the Leagues Cup title in August when Messi lit up the tournament and collected his first trophy since joining the club in July.
Saturday's loss was the second straight for Miami following their 4-1 crushing at Chicago on Wednesday and Martino's team have won just once in their last six MLS games.
Messi had played in the only win in that stretch, coming off in the first half of their 4-0 win over bottom club Toronto.
After the Leagues Cup triumph in Nashville on August 19th, there was talk of Miami adding two more trophies this season but, without Messi, they lost their US Open Cup final to Houston and then faded out of contention in the league.
"Honestly, I expected the season to be exactly the opposite of what happened," said Martino, who took over just before Messi's arrival.
"My expectations were to put the group together, shape it football-wise, not transcend too much in the Leagues Cup, compete well in the Cup (US Open Cup) and qualify for the play-offs, but it was exactly the opposite as it usually happens when a coach thinks something," he said.
"Out of three possible tournaments we have won one, it is a very important fact for us because the league has 28 other teams. Houston and we won, whoever wins the league will be the third winner and there will be 25 others who will not have won. From that point of view, we're happy."
Miami's final two games are both against Charlotte with the last on the road on October 21st. It remains to be see if Messi, who has been called up for Argentina's upcoming World Cup qualifiers, will feature.
Rooney exits
Rooney's DC United failed to quality despite beating New York City FC 2-0 in their final game of the season and within minutes the former Manchester United forward's exit was announced.
"It's the right time. I've done everything I can to get this club into the play-offs. It's not a single thing that's happened. It's about timing," said Rooney.
In the Western Conference, five-time MLS champions Los Angeles Galaxy failed to make the play-offs after a 5-2 defeat to Minnesota United.
Finnish striker Teemu Pukki scored four second-half goals for Minnesota who had fired their English coach Adrian Heath on Friday.
Austin, beaten by Los Angeles FC in the Western Conference final last year, suffered another disappointment at the hands of LAFC with their 4-2 loss at home to the champions ending their post-season hopes.
Gabon international Denis Bouanga scored twice for LAFC to move clear as the league's leading scorer with 19 goals.
US Open Cup winners Houston boosted their hopes of a top-four finish and home-field advantage in the first round of the play-offs with Nelson Quinones grabbing a brace in a 5-1 thrashing of Colorado.