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<p>Two-dimensional cultures and animal models have long been used in cancer research, but neither of the two systems can preserve tumor architecture, diffusion gradients, and microenvironmental constraints. Three-dimensional (3D) tumor spheroids overcome such shortcomings by allowing spatial analysis of drug penetration, cellular heterogeneity, and treatment resistance. This review describes significant strategies of spheroid fabrication, such as scaffold-based systems, hanging-drop techniques, liquid overlay techniques, and microfluidic platforms. Their use in theranostics is presented in the fields of nuclear medicine, nanocarrier-based delivery, photodynamic therapy, and immunotherapy. The critical analysis reveals the importance of such determinants of performance as the density of extracellular matrix, hypoxia tolerance, diffusion of a nanoparticle, and localization in the intracellular space. Chronic difficulties are inter-spheroid variability, lack of complete immune representation, and the inability to quantitatively assess penetration and dose distribution. Future directions include standardization of protocols, microfluidics and artificial intelligence interfaces, and designing patient-derived spheroid models of precision oncology. Altogether, 3D spheroids represent an accessible, biologically relevant, and scalable platform between accessible in vitro systems and clinical translation.</p>

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Theranostic nanoplatforms applications of 3D tumor spheroid models: progress, limitations, and future directions

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月31日

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  • Shalini Singh Negi,
  • Devendra Singh

摘要

Two-dimensional cultures and animal models have long been used in cancer research, but neither of the two systems can preserve tumor architecture, diffusion gradients, and microenvironmental constraints. Three-dimensional (3D) tumor spheroids overcome such shortcomings by allowing spatial analysis of drug penetration, cellular heterogeneity, and treatment resistance. This review describes significant strategies of spheroid fabrication, such as scaffold-based systems, hanging-drop techniques, liquid overlay techniques, and microfluidic platforms. Their use in theranostics is presented in the fields of nuclear medicine, nanocarrier-based delivery, photodynamic therapy, and immunotherapy. The critical analysis reveals the importance of such determinants of performance as the density of extracellular matrix, hypoxia tolerance, diffusion of a nanoparticle, and localization in the intracellular space. Chronic difficulties are inter-spheroid variability, lack of complete immune representation, and the inability to quantitatively assess penetration and dose distribution. Future directions include standardization of protocols, microfluidics and artificial intelligence interfaces, and designing patient-derived spheroid models of precision oncology. Altogether, 3D spheroids represent an accessible, biologically relevant, and scalable platform between accessible in vitro systems and clinical translation.