On the basis of the predictions of statistical-thermodynamic models, it is postulated that excluded volume effects may play a significant role in the stability, interaction, and function of proteins. We studied the effects of confinement on protein un/refolding and stability. Our approach was to encapsulate a model protein, RNase A, in a mesoporous silica, MCM-48, with glasslike wall structure and with well-defined pores to create a crowded microenvironment. To the best of our knowledge, this is the first report where pressure perturbation and differential scanning calorimetric techniques are employed to evaluate the stability, hydration, and volumetric properties of the confined protein. A drastic increase in protein stability ( approximately 30 degrees C increase in unfolding temperature) is observed. The increase in stability is probably not only due to a restriction in conformational space (excluded volume effect due to nonspecific interactions) but also due to an increased strength of hydration of the protein within the narrow silica pores.