Kew Jaliens is in danger of missing the World Cup finals after picking up a leg injury on Thursday.
The AZ Alkmaar defender was named in Marco van Basten's 28-man pre-squad and had high hopes of heading to Germany.
He was named in the staring line-up as van Basten's side took on a strong Dutch Under-21 side.
Jaliens looked composed for over 70 minutes, but was forced off 13 minutes from time.
He left the field with what looked like a serious injury and his hopes of heading to Germany are in the balance.
The contest ended 0-0, with Ruud van Nistelrooy an interested spectator.
Holland coach Van Basten opted to leave his star striker on the bench, following his turbulent week at Manchester United.
Dutch national team: Timmer (Stekelenburg/46), Jaliens (Mathijsen/77), Heitinga, Opdam (Vennegoor of Hesselink/68), De Cler, Landzaat, Sneijder, Cocu (Kuijt/46), Castelen, Huntelaar, Babel