The mechanism by which cryosolvents such as alcohols modify and penetrate cell membranes as a function of their concentration and hydration state remains poorly understood. We conducted molecular dynamics simulations of 1,2-dioleoyl-sn-glycero-3-phosphocholine bilayers in the presence of aqueous solutions of four common penetrating hydroxylated cryosolvents (methanol, ethylene glycol, propylene glycol, and glycerol) at varying concentration ranges and across three different hydration states. All cryosolvents were found to preferentially replace water at the bilayer interface, and a reduction in hydration state correlates with a higher proportion of cryosolvent at the interface for relative concentrations. Minor differences in chemical structure had a profound effect on cryosolvent-membrane interactions, as the lone methyl groups of methanol and propylene glycol enhanced their membrane localization and penetration, but with increasing concentrations acted to destabilize the membrane structure in a process heightened at higher hydration states. By contrast, ethylene glycol and glycerol promoted and retained membrane structural integrity by forming hydrogen-bonded lipid bridges via distally located hydroxyl groups. Glycerol exhibited the highest capacity to cross-link lipids at relative concentrations, as well as promoted a bilayer structure consistent with a fully hydrated bilayer in the absence of cryosolvent for all hydration states investigated.