The aim of this study was analyze the effect of playing against biological matched and un-matched opposition, on physical performance and spatial exploration behavior of youth basketball players. Thirty under-14 to 16 basketball players were assigned to different teams according to maturity status (Pre-, Mid-, and Post-Peak Height Velocity [PHV]), and participated in basketball matches against matched (same maturity status), and un-matched (different maturity status) opposition. Maturity status was estimated considering the percentage of predicted adult height. Workload data was collected via inertial devices (IMUs) and Ultra-Wide Band (UWB)-based system. Heart rate was recorded with individual HR monitors. The Pre-PHV performed significantly more accelerations and decelerations and explored more space against matched opposition. Against un-matched opposition, the Pre-PHV presented higher average speed, body impacts, and Player Load. Both Mid- and Post-PHV covered more distance against matched opposition than against Pre-PHV. Games against Pre-PHV involved lower distance covered, average speed, Player Load, and higher accelerations and decelerations, than against Mid- and Post-PHV. The Pre-PHV athletes performed a higher number of accelerations and decelerations comparing to the Mid and Post-PHV players. Also, a significant interaction effect (group x time) was found in distance covered, average speed, body impacts, and Player Load. The type of opposition influenced physical performance and spatial exploration behavior during basketball matches, particularly of less-mature players. Based on present findings, practitioners can select the most suitable game format, considering the physical, technical, tactical, and psychological development needs, individualizing training stimulus.