Bolton boss Phil Parkinson says he contacted the English Football League before Saturday's 5-0 loss at Tranmere over welfare concerns after playing youth players in every game this term.
Just three senior outfield players were fit for the trip to Prenton Park.
Wanderers, who are in administration, are currently awaiting the completion of a takeover by Football Ventures.
"I've spoken many times in the past couple of weeks. Asking young players to go again is not right," he said.
"We sent an email to the English Football League explaining about our concerns over the welfare of our young players playing these games in quick succession, but we were told we had to play the game."
When contacted by BBC Sport, the EFL said welfare checks are in place for any under-18s playing first-team football.
Bolton's first game of the season against Wycombe had been under threat of suspension until the EFL confirmed that Bolton had provided sufficient evidence of financial viability.
The club, who cancelled three pre-season friendlies after players claimed they went five months without payment, were allowed to sign James Weir and Josh Earl prior to their opening game.
However, Parkinson has filled their starting line-ups with youth players in each game this season, including fielding the youngest team in their history in their goalless draw against Coventry on 10 August.
Bolton's under-23 game against Colchester, which had been scheduled to be played on Monday, has since been rearranged.
Speaking to BBC Radio Manchester, Parkinson continued: "The combination of people not getting this deal to sell the club over the line and the tangled web which that is involved in, and making us play the game [against Tranmere], has hung all the staff and players out to dry.
"Honestly, it's just unacceptable. This is Bolton Wanderers Football Club, a fantastic club with a great history, and we need a bit of help. If it's not going to be sold, we need to be able to bring some players in."
The takeover by Football Ventures had been blocked by a court order, issued by former Watford owner Laurence Bassini, who had tried to buy the club earlier this year before the deal fell through.
The sale now appears ready to go through, however, after the order was adjourned in court on Wednesday.
Parkinson added: "If you go back to the Wycombe game (on 3 August), you thought 'could we put a team on the pitch with the situation we're in?' and we did. Now it's two weeks later and we're in the same situation.
"No new players in, no sign of the deal getting done, there's signs of people out there wanting to buy the club but still no sign of it going over the line."