As a theoretical basis for treatment of carpal tunnel syndrome (CTS) and expanding upon part 1 of this study, the authors investigated the effects of static loading (weights) and dynamic loading (osteopathic manipulation [OM]) on 20 cadaver limbs (10 male, 10 female). This larger study group allowed for comparative analysis of results by sex and reversal of sequencing for testing protocols. In static loading, 10-newton loads were applied to metal pins inserted into carpal bones. In dynamic loading, the OM maneuvers used were those currently used in clinical settings to treat patients with CTS. Transverse carpal ligament (TCL) response was observed by measuring changes in the width of the transverse carpal arch (TCA) with three-dimensional video analysis and precision calipers. Results demonstrated maximal TCL elongation of 13% (3.7 mm) with a residual elongation after recovery of 9% (2.6 mm) from weight loads in the female cadaver limbs, compared to less than 1 mm as noted in part 1, which used lower weight loads and combined results from both sexes. Favorable responses to all interventions were more significant among female cadaver limbs. Higher weight loads also caused more linear translatory motion through the metal pins, resulting in TCA widening equal to 63% of the increases occurring at skin level, compared to only 38% with lower loads. When OM was performed first, it led to greater widening of the TCA and lengthening of the TCL during the weight loading that followed. Both methods hold promise to favorably impact the course of management of CTS, particularly in women.