The design and synthesis of biopolymer nano-and micro-formulations are a new trend with growing priority in scientific research and development in the fields of biomedicine, bioorganic/ medicinal chemistry, pharmaceutics, agrochemistry and food industry. This incorporates a vast variety of improved and newly-developed analytical and physico-chemical techniques for green and efficient synthesis. The scope of the present review was to outline the recent advances in the methods for design of novel chitosan micro-and nano-carriers and transporters. Special emphasis is laid on their functionalities and capacity for encapsulation of natural bioactive compounds and controlled in vitro/in vivo release in various biological/physiological media. Expectations for the application of chitosan formulations and chitosan-based hybrid systems are progressively increasing as the knowledge regarding their physical, chemical and biological properties constantly expands. Thus, this review proposes insights on the objective assessment of the capacity, applicability and versatility of newly-designed chitosan-based hybrid systems. A detailed integrative approach, which incorporates the innovative scientific achievements based on complex novel, precise and reliable analytical procedures and methods for qualitative and quantitative morphological, structural, spectral, chemical and biochemical analyses of the bioprecursors and the designed chitosan-carrier micro/nano-hybrid systems, is applied. Sustainable knowledge on the mechanism and methods of natural bioactive substances encapsulation and in vitro/in vivo release is reviewed and discussed.