Basic principles of laser pulses interaction with solid and liquid materials using advanced pulsed laser technologies are introduced and new recent results in synthesis of biomaterial layers are reviewed. The layers are optimized based upon the results of physical-chemical investigations, while biocompatibility, bioactivity and biodegradation were assessed by dedicated in-vitro tests.  The coating of metallic implants with composite alendronate (AL)-HA by MAPLE was demonstrated to enhance human osteoblasts proliferation and differentiation, while inhibiting osteoclasts growth, with benefic effects for the treatment of bone diseases. MAPLE thin films of calcium alendronate monohydrate, octacalcium phosphate, and CaAL*H2O / OCP composite on Titanium substrates proved that the presence of calcium alendronate in the coatings dramatically inhibits proliferation, differentiation and activity of osteoporotic osteoclast. The bisphosphonate promotes osteoblast viability and activity. Osteoblast differentiation is enhanced when the coating is a composite of CaAL*H2O and OCP, most likely due to the good bioactivity of OCP.  Structures with compositional gradient are obtained by Combinatorial-MAPLE (C-MAPLE) by the simultaneous laser vaporization of two targets. Synchronized MAPLE of levan and oxidized levan cryogenic targets is applied in order to transfer under protection and assemble a two-compound biopolymer film structure. FTIR micro-spectroscopy confirmed the existence of a composition gradient along the length of the sample. In-vitro cell culture assays illustrate characteristic responses of cells to specific surface locations. Cells attached along the gradient are in direct proportion with oxidized levan concentration.  C-MAPLE is applied to synthesize crystalline gradient thin films of Sr-substituted hydroxyapatite (Sr-HA) and Zolendronate modified hydroxyapatite (ZOL-HA). The inhibitory action of ZOL on osteoclast viability and activity is more efficient than that of Sr, which has a greater beneficial role on osteoblast proliferation and viability. C-MAPLE allows to modulate the composition of the thin films and hence the promotion of bone growth and the inhibition of bone resorption.  Our conclusion is that the thin films prepared by pulsed laser techniques are identical in chemical composition, structure, morphology, and most likely functionality resembling the base material, as proved by physical-chemical characterization and in-vitro testing. C-MAPLE opens the possibility to combine and immobilize two or more organic materials on a substrate in a well defined manner by laser evaporation under protection.