Multifunctional 3D‐printed nanocomposites based on poly(lactic‐co‐glycolic acid), that is, PLGA (RESOMER® LG857S) were developed for simultaneous monitoring of cells and scaffold as a function of time and spectral responses. These were achieved by impregnating carbon quantum dots (CQDs) on PLGA using melt‐blending, plasticating extrusion, and 3D‐printing. The nanocomposites enabled enhanced bio‐affinity and cellular interactions for bone tissue engineering (TE). PLGA (control) and PLGA‐CQD scaffolds were used for growing human adipose‐derived‐stem‐cells (ADSCs) and tested for cell biocompatibility, cellular adhesion, growth, and osteogenesis. CQDs were found to enhance the hydrophilicity of nanocomposites and promote cellular nesting. MTS assays confirmed that CQDs on PLGA act as cell anchoring sites, thereby enhancing seeding efficiency and cell proliferation. Alkaline phosphate tests showed increased osteogenesis and Alizarin assays confirmed enhanced bone mineralization on PLGA‐CQD. The qPCR tests based on selected mRNA expressions showed that the incorporation of CQDs significantly enhanced osteogenesis of ADSCs during all three phases of cell differentiation. The intrinsic luminescence of the composites allowed label‐free monitoring of cell proliferation and bone mineralization on the scaffolds. Thus, the CQDs facilitated significant enhancements in composite processability with customized fabrication of 3D printed scaffolds, bone tissue osteoconductivity, and monitoring of cell‐scaffold activities, offering multifunctional benefits for bone TE.
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