Metabolic-dysfunction-associated steatotic liver disease (MASLD) has emerged as a significant public health concern, attributed to its increasing prevalence and correlation with metabolic disorders, including obesity and type 2 diabetes. Recent research has highlighted that mitochondrial dysfunction can result in the accumulation of lipids in non-adipose tissues, as well as increased oxidative stress and inflammation. These factors are crucial in advancing the progression of MASLD. Despite advances in the understanding of MASLD pathophysiology, challenges remain in identifying effective therapeutic strategies targeting mitochondrial dysfunction. This review aims to consolidate current knowledge on how mitochondrial imbalance affects the development and progression of MASLD, while addressing existing research gaps and potential avenues for future research. This review was conducted after a systematic search of comprehensive academic databases such as PubMed, Embase, and Web of Science to gather information on mitochondrial dysfunction as well as mitochondrial-based treatments for MASLD.

The construction of highly biomimetic in vitro tissue barrier models is essential for new drug development, as it enables accurate evaluation of the permeability efficiency, safety, and pharmacokinetic properties of drug candidates. Animal models have long been regarded as the standard criterion for assessing drug efficacy; however, they pose increasingly severe ethical concerns. Moreover, substantial interspecies differences in transmembrane transporter expression and metabolic pathways contribute to the low rate of successful clinical translation. Traditional 2D cell culture models lack a 3D matrix microenvironment and physiological mechanical signals similar to those in vivo. Consequently, epithelial or endothelial cells are prone to phenotypic alteration. They struggle to assemble structurally intact and functionally mature tight barriers. As a result, drug permeability and toxicity results often deviate markedly from in vivo physiological conditions. In contrast, electrospinning technology offers a promising approach for constructing biomimetic barriers. Electrospun nanofiber membranes closely resemble the fibrous network structure of the natural extracellular matrix and provide several distinct advantages, including precisely controllable pore sizes, large specific surface areas, and readily modifiable functional surfaces. Furthermore, the arrangement and physicochemical properties of the fibers can be precisely tailored to provide cells with inductive cues that mimic the in vivo physiological microenvironment. This review summarizes the design strategies of electrospun nanofiber membranes for the construction of key tissue barriers, including the lung, blood–brain barrier, intestine, kidney, and skin. In addition, it discusses recent advances in the application of these membranes for drug screening. Finally, universal design principles for cross-barrier construction are proposed. Overall, this review aims to support the development of more efficient and reliable in vitro drug-screening platforms.