The thermodynamics of temperature-responsive polymeric layers was analyzed using a two-state coil to globule model to which the van't Hoff relationship was applied. For soluble homopolymer poly(N-isopropylacrylamide (pNIPAAm), enthalpies of transition, Delta H(vH), were calculated using varations in ultraviolet-visible (UV-vis) spectroscopy with temperature to be 8400 +/- 30 and 1652 +/- 4 kJ mol-cooperative unit(-1) for standard synthesis and initiated chemical vapor deposition (iCVD), respectively. For the insoluble surface-bound layer of cross-linked iCVD poly(N-isopropylacrylamide-co-di(ethylene glycol) divinyl ether) [p(NIPAAm-co-DEGDVE)], Delta H(vH) was determined to be 810 +/- 30 kJ mol-cooperative unit(-1) using quartz crystal microbalance with dissipation monitoring (QCM-D). Microcalorimetry measurements showed the enthalpies per mole NIPAAm monomer to be 5.8 +/- 0.2, 3.5 +/- 0.6, and 3.1 +/- 0.3 kJ mol-NIPAAm(-1), resulting in cooperative unit sizes of 1460 +/- 60, 470 +/- 80, and 260 +/- 30 monomer units for the standard pNIPAAm, iCVD pNIPAAm, and p(NIPAAm-co-DEGDVE) systems, respectively. These values indicate that both per monomer enthalpic contribution as well as cooperative unit size are primary factors contributing to the variations in van't Hoff enthalpies for the three systems studied. Diffusion of bovine serum albumin (BSA) into swollen p(NIPAAm-co-DEGDVE) films below its lower critical solution temperature was elucidated via QCM-D measurements. These data provided a calculated diffusion coefficient of (3.5 +/- 0.1) x 10(-14) cm(2) s(-1) of BSA into the swollen hydrogel film with a mesh size of 6.0 +/- 0.2 nm (compared to the hydrodynamic radius of BSA, rH = 3.36 nm).