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Biomechanical Contrast Between Native and Decellularized Triple-Negative Breast Tumors in Mice

Authors :

Mohammad Javad Farjam¹ Saman Asadi² Ashkan Azimzadeh³ Saeid Amanpour⁴ AbdolMohammad Kajbafzadeh⁵ Mohammad Ali Nazari⁶

1- دانشگاه تهران 2- دانشگاه علوم پزشکی تهران 3- دانشگاه علوم پزشکی تهران 4- دانشگاه علوم پزشکی تهران 5- دانشگاه علوم پزشکی تهران 6- دانشگاه تهران

Keywords :

4T1 tumor،(triple-negative breast cancer (TNBC،decellularization،(extracellular matrix (ECM،atomic force microscopy (AFM)،tissue stiffness،tumor microenvironment

Abstract :

Triple-negative breast cancer (TNBC) is an aggressive subtype that does not have any targeted receptor-based therapies available, for which the 4T1 cell line–based murine model hosted in BALB/c (“Bagg Albino”) mice offers a viable syngeneic model. In this work, we provided a comparative assessment of the structural and mechanical properties of native and decellularized tumors induced by the 4T1 cell line (hereafter referred to as 4T1 tumors). Hematoxylin and eosin (H&E) images visually confirmed successful removal of nuclear content with maintenance of the architecture of the cross-linking throughout the extracellular matrix (ECM). Scanning electron microscopy (SEM) displayed hierarchical ultrastructure, with fibrous and porous morphology preserved after decellularization. Atomic force microscopy (AFM) nano-indentation testing illustrated a significant reduction in stiffness following decellularization: the native 4T1 tumor exhibited a mean Young’s modulus of 71.3 ± 19.7 kPa, whereas the decellularized ECM measured 10.5 ± 8.6 kPa. This ~7-fold decrease highlights the role of cellular components in conferring tumor rigidity while confirming the preservation of ECM scaffolds. The normality test (Lilliefors) showed p = 0.0181 for native tumor samples and p = 0.0010 for decellularized samples, indicating that data may not be normally distributed (p < 0.05). AFM indentation was performed using a quadratic pyramid probe. Tumors were generated in 8 BALB/c mice, of which 4 samples were subsequently decellularized for analysis. These findings validate the feasibility of decellularized 4T1 tumors as biomimetic scaffolds for studying tumor–stroma interactions, mechanobiology, and therapeutic modeling.