Purpose: To investigate the effects of anterior guidance (AG) change on the working (WCP) and non-working condylar paths (NWCP), and lower incisor path (LIP) using a splint with flat (FAG) or steep AG (SAG). Materials and Methods: The samples consisted of six young adult women (mean age=$23.5{pm}3.3$ years). Inclusion criteria were skeletal Class I and normodivergent pattern, normal overbite/overjet, minimal slide from retruded cuspal position to intercuspal position, no temporomandibular disorder signs and symptoms, mutually protected occlusion, and minimal tooth wear. After the values of natural AG (NAG) were obtained as a reference for each patient, two types of splints ($15^{circ}$ flatter and steeper than NAG) were made. After insertion of the splints with FAG or SAG, the WCP, NWCP, and LIP were recorded five times for each patient using an ultrasonic AQR (SAM, Munich, Germany) and statistical analysis was subsequently performed. Result: NAG exhibited postero-superior movement in the WCP and did not show a noticeable immediate side shift (ISS) or difference between the eccentric (EP) and returning paths (RP) in the NWCP. FAG was associated with an irregular and excessive WCP, an increase in ISS, and a difference between EP and RP in the NWCP. SAG showed minimal WCP movement and a decrease in the extent of difference between EP and RP in the NWCP. LIP showed significant differences in EP and in RP (P<0.001, all; FAG<NAG<SAG). Conclusion: Since AG change can affect the WCP, NWCP, and LIP, it is necessary to establish proper AG during orthodontic treatment.