Advanced fabrication of next-generation functional foods: integrating beneficial life forms through 5D food printing

Distinct from traditional food printing in their utilization of beneficial life forms (BLFs), this review firstly concludes and analyzes an innovative five‑dimensional food printing (5DFP) approach. In this approach, BLFs embedded in an edible printing matrix (3D) undergo time‑dependent (4D) growth, metabolism, or differentiation, actively driving the dynamic evolution of matrix structure, function, and sensory properties. It also simultaneously generating multiple signals (e.g., color, flavor, shape and nutrient changes) that can be visually identified or amplified for detection, thereby facilitating data collection and feedback‑driven printing optimization. The sources and features of BLFs for 5DFP are systematically summarized, including applied natural fermentative microorganisms and promising photosynthetic microorganisms, plant/animal functional cells, and their engineered variants, showing promise for nutrient synthesis, targeted delivery, and living label applications. Moreover, optimization strategies such as BLFs modulation, carrier material and printing parameter optimization, and multi‑nozzle systems integrated with physical field‑assisted technologies are discussed for feasibility validation. Moving beyond these technical aspects, key barriers including biosafety, regulatory approval, scalability, ethics, consumer acceptance, and long‑term stability are also critically highlighted. Accordingly, future research should focus on expanding BLF materials, advancing technical strategies, and reinforcing regulatory systems, while AI‑based approaches remain a promising yet exploratory direction in 5DFP.