Various applications within last decades such as bacterially resistant surfaces, soft robotics, drug delivery systems, sensors and tissue engineering are poised to feature the importance of the ability to control bio-interfacial interactions. An enhanced attention is dedicated to designing smart stimuli-responsive interfaces for DNA, drug delivery, protein and cell based applications. Within this context, the thermoresponsive materials, especially poly(N-isopropylacrylamide) (pNIPAm) have been intensively used in tissue engineering applications for a controlled detachment of proteins and cells with a minimum of invasive effect on protein and cell structural conformation. The properties of smart bio-interfaces can be controlled by its composition and polymer architecture. Therefore, appropriate methods for obtaining controlled coatings are necessary. Laser methods were successfully used in the last decades for obtaining controlled organic and inorganic coatings for various types of applications, from electronics to tissue engineering. Among these, Matrix-Assisted Pulsed Laser Evaporation (MAPLE) technique bring us a step forward to other laser methods by avoiding damage and photochemical decomposition of materials. In this chapter we describe materials and approaches used for design of smart bio-interfaces aimed at controlling protein and cells behavior in vitro, focusing MAPLE method for tuning coatings characteristics in relation with biological response.