Fig. 4: Transplantation of neuromuscular tissues fabricated by acoustofluidic bioassembly method for volumetric muscle loss.

a Schematic diagram and timeline of production and implantation of 3D neuromuscular tissue generated by acoustofluidic bioassembly (AB). b Schematic of muscle fibers in the defect, regenerated region, and host region in the cross-section of skeletal muscle. In normal skeletal muscle (host), the nuclei of muscle fibers are located at the periphery of the muscle fiber. Myofibers with central nuclei are observed in the regenerated region. c Representative images of Masson’s trichrome (MT) and Hematoxylin and eosin (H&E) stained cross-sections of volumetric muscle loss (VML)-injured quadriceps muscles at 6 and 12 weeks after. Black dotted lines in gross view represent the host regions before ablation. Scale bars in gross view and microscopic image represent 500 μm and 20 μm, respectively. d–f Quantitative analysis of the number of centralized nuclei (d), fibrotic area (e), and the average cross-sectional area (CSA) of myofiber (f) based on MT staining images (n = 5 biologically independent samples). g Similarity of the histograms of CSA of myofibers of gel only, iMPC+MN R, iMPC AB, or iMPC+MN AB groups versus (vs.) those of NT or normal groups. All data are presented as mean ± S.D., and statistical differences were determined with one-way ANOVA followed by Tukey’s test. #P < 0.05 and ##P < 0.01 versus NT, $P < 0.05 and $$P < 0.01 versus gel, ^^P < 0.01 versus iMPC+MN R, and *P < 0.05 and **P < 0.01 versus Normal. Source data are provided as a Source Data file.