bims-mitrat 2026-08-02 papers

J Orthop Translat. 2026 Jul;59 101171

Hypoxia-preconditioned mitochondrial transplantation multidirectionally modulates Schwann cell functions to repair peripheral nerve injury in rats.

Jin Dong, Bing Yan, Xushen Zhao, Jinyu Bai, Huajian Shan, Xiang Gao, Lei Sheng, Jun Dai, Fengxian Jiang, Mingchao Zhang, Chaowen Bai, Xiaozhong Zhou, Shuai Wei.

Background: Peripheral nerve injury (PNI) often leads to sensory and motor dysfunction. This study investigated the promoting effect of hypoxia-preconditioned mitochondrial transplantation on the structural and functional reconstruction of peripheral nerves and the molecular mechanisms involved.
Methods: We explored the effects of hypoxia-preconditioned mitochondrial transplantation on Schwann cell (SC) phenotypes, including proliferation, migration, cellular senescence, and mitochondrial membrane potential, and explored the underlying molecular mechanism through Western blotting. We assessed axonal and myelin regeneration, as well as motor function recovery, in injured rats through behavioral tests, morphological analysis, and electrophysiological detection.
Results: Hypoxia preconditioning significantly increased the mitochondrial membrane potential in SCs without altering their ultrastructure or the normal expression of the COX IV and TOMM20 proteins. The uptake efficiency of SCs for exogenous mitochondria was significantly greater than that of neurons, endothelial cells, fibroblasts, and other cells. Hypoxia-preconditioned mitochondrial transplantation activated the MAPK (Ras-ERK-c-Fos/c-Jun) pathway; promoted SC proliferation, migration, and dedifferentiation; inhibited H2O2-induced cellular senescence; and partially restored the mitochondrial membrane potential. Hypoxia-preconditioned mitochondrial transplantation significantly accelerated axonal growth, promoted axonal remyelination, led to significant recovery of electrophysiological function, and improved motor function.
Conclusion: Hypoxia preconditioning enhances the mitochondrial membrane potential of SC-derived mitochondria. Mitochondrial transplantation modulates SC functions by activating the Ras-ERK-c-Fos/c-Jun pathway, significantly increases nerve fiber density and myelin thickness, alleviates gastrocnemius muscle atrophy, and promotes motor function recovery, providing a new strategy for the clinical treatment of PNI.
Translational potential: We established a hypoxia-preconditioned mitochondrial transplantation method that modulates SC functions, accelerates axonal and myelin regeneration, and promotes motor function recovery. This study provides a novel clinical strategy for treating peripheral nervous system injuries.

Keywords: Dedifferentiation; Hypoxia; Mitochondrial transplantation; Nerve regeneration; Peripheral nerve injury; Schwann cell

DOI: https://doi.org/10.1016/j.jot.2026.101171

Cells. 2026 Jul 16. pii: 1279. [Epub ahead of print]15(14):

Optimized Isolation and Cryopreservation of Functional Mitochondria for Transplantation and Therapeutic Applications.

Vikky Awasthi, Aasthika Das, Meriem Bkhache, Abeer Alshambky, Glenn S Gerhard, Karim Bahmed, Timothy Cashman, Rihab Bouchareb.

Mitochondria play a central role in numerous physiological and pathological processes, and mitochondrial transplantation is emerging as a promising strategy to restore cellular function and mitigate disease. The success of this approach depends critically on the methods used to isolate, preserve, and retrieve intact, functional mitochondria. Objective: To optimize an isolation strategy that preserves mitochondrial integrity, dynamics, and metabolic activity and to evaluate conditions that enable short-term storage for future organelle biobanking applications. Methods: We compared a mitochondria isolation method developed in our laboratory (Protocol A) with a commercially available kit (Protocol B). Donor mitochondria were isolated from proximal tubular cells and transplanted into HEK293T recipient cells. Mitochondrial functionality was assessed following transfer into HEK293T cells by measuring reactive oxygen species (MitoSOX Red), oxygen consumption rate (OCR) using Seahorse XF analysis, and high-resolution imaging of mitochondrial morphology and dynamics. We further evaluated mitochondrial storage at low temperature and subsequent functional recovery. Results: Protocol A enabled faster isolation (~30 min) than Protocol B (~80 min) and yielded mitochondria with higher transplantation efficiency, greater OCR, preserved dynamic morphology, and lower oxidative stress. Mitochondria isolated using Protocol A remained metabolically active after transplantation and continued to exhibit fission and fusion, whereas those isolated using Protocol B showed reduced dynamic behavior. Importantly, mitochondria isolated with Protocol A retained functional integrity after low-temperature storage, supporting their potential for standardized preservation. Conclusions: This study presents a robust, efficient, and reproducible isolation and frozen-storage protocol that yields highly functional mitochondria suitable for transplantation. The ability to preserve mitochondrial function after storage further highlights the potential for developing organelle biobanks to support future research and therapeutic applications.

Keywords: cryopreservation of mitochondria; mitochondria isolation; mitochondria transplantation

DOI: https://doi.org/10.3390/cells15141279

Adv Healthc Mater. 2026 Jul 28. e71495

Source-Dependent Mitochondrial Transplantation Drives Schwann Cell Bioenergetic Reprogramming and Peripheral Nerve Regeneration.

Xiangling Li, Yunjie Huang, Xiaojian Cao, Hongmei Luo, Yuan Zou, Fatima Aldali, Sheng Chen, Yajie Li, Qiuzhi Zhou, Hong Chen.

Following peripheral nerve injury (PNI), metabolic reprogramming of Schwann cells (SC) plays a critical role in axonal regeneration and functional recovery. Although mitochondrial transplantation (MT) has been proposed as a potential therapeutic strategy, it remains unclear whether its effect is limited to transient energy supplementation. We systematically compared the differential effects of mitochondria derived from human induced pluripotent stem cells (iPSCs, iP-MT) and induced mesenchymal stem cells (iMSCs, iM-MT) on regulating SC metabolism and promoting peripheral nerve regeneration. The results showed that exogenous mitochondria were efficiently taken up by SCs, not only increasing ATP production and mitochondrial membrane potential but also reshaping the metabolic distribution between glycolysis and oxidative phosphorylation. This remodeling promoted cell proliferation, migration, and neurotrophic function, while enhancing antioxidant capacity. In a rat sciatic nerve injury model, MT significantly improved axonal regeneration, remyelination, and the recovery of sensory and motor functions. Notably, iM-MT demonstrated stronger metabolic adaptability and therapeutic efficacy. MT reduced ROS accumulation, increased ATP production, and attenuated apoptosis under oxidative stress. This study reveals that MT promotes nerve repair by driving metabolic reprogramming rather than merely providing temporary energy, offering a new theoretical basis for optimizing the selection of mitochondrial donors.

Keywords: glycolysis; mitochondrial biogenesis; mitochondrial transplantation; oxidative stress; peripheral nerve regeneration

DOI: https://doi.org/10.1002/adhm.71495

Biology (Basel). 2026 Jul 09. pii: 1112. [Epub ahead of print]15(14):

Mitoception: A Novel Strategy to Alleviate Pulmonary Fibrosis.

Sarayu Bhogoju, Parth Patel, Neeraj Kapur, Prashant D Kunjadia, Ajoy Aloysius, Dave-Preston Esoe, Jamie L Sturgill, Christine F Brainson, Luksana Chaiswing, Patrick G Sullivan, Anthony N Gerber, Edward Castillo, Stewart F Graham, Ishanu Chattopadhyay, Girish Nair.

Pulmonary fibrosis (PF) is a progressive lung condition characterized by irreversible scarring and high mortality, with limited effective treatments. Mitochondrial dysfunction has emerged as a critical factor in fibroblast activation in PF, although approaches to restore mitochondrial function remain underexplored. The present study investigated whether mitochondrial transfer from alveolar type II epithelial cells (A549) to patient-derived fibroblasts could restore mitochondrial function and bioenergetics. Histological analysis of fibrotic lungs reveals increased collagen deposition and elevated profibrotic markers, accompanied by reduced expression of mitochondrial biogenesis and respiratory proteins compared to non-fibrotic controls, indicating mitochondrial impairment. Freshly isolated donor mitochondria were functionally validated before mitoception using Seahorse analysis and patient-derived fibroblasts were confirmed by qRT-PCR using fibroblast-specific markers. In vitro transfer of mitochondria to diseased patient-derived fibroblasts exhibited a modest, dose and time-dependent increase in mitochondrial membrane potential compared to normal fibroblasts. Gene expression analysis revealed decreased fibrosis-associated markers and increased expression of mitochondrial and antioxidant genes following mitoception. Seahorse analysis after mitoception revealed enhanced ATP-linked respiration and improved selected mitochondrial bioenergetic parameters, whereas maximal respiration and spare respiratory capacity demonstrated variable responses. In contrast, normal fibroblasts displayed minimal changes. Collectively, these findings indicate that mitochondrial transfer modulates fibroblast bioenergetics and profibrotic signaling, supporting its potential as a therapeutic strategy for pulmonary fibrosis.

Keywords: mitoception; mitochondrial dysfunction; mitochondrial transfer; profibrotic signaling; pulmonary fibrosis (PF)

DOI: https://doi.org/10.3390/biology15141112

Bio Protoc. 2026 Jul 20. 16(14): e5744

Preparation and Characterization of Neutrophil Membrane-Fused Mitochondria (nMITO).

Qing Zhang, Yuqin Yue, Xing Zhou.

Mitochondrial transplantation is an emerging strategy for cellular repair, yet its efficiency is often limited by poor targeting and environmental instability. This protocol details the fabrication and comprehensive characterization of neutrophil membrane-fused mitochondria (nMITO), a hybrid organelle platform designed to combine the metabolic vigor of natural mitochondria with the targeting and anti-inflammatory properties of neutrophil membranes. We describe an optimized workflow for mouse heart mitochondrial isolation, lipopolysaccharide (LPS)-activated neutrophil membrane (NEM) extraction, and the subsequent sonication-mediated fusion process. Characterization techniques include dynamic light scattering (DLS) for size and zeta potential, transmission electron microscopy (TEM) for ultrastructural integrity, and bioenergetic assays [ATP synthesis and tetramethylrhodamine methyl ester (TMRM)-based membrane potential] to ensure functional preservation. Key features • The protocol provides a methodology for the isolation of neutrophil membranes from mouse bone marrow and mitochondria from the heart. • The protocol provides a methodology for the fabrication of neutrophil membrane-fused mitochondria (nMITO).

Keywords: Membrane coating; Mitochondria isolation; Mitochondrial assessment; Neutrophil membranes; nMITO

DOI: https://doi.org/10.21769/BioProtoc.5744