Human skeletal muscle tissue chip autonomous payload reveals changes in fiber type and metabolic gene expression due to spaceflight.

Microphysiological systems provide the opportunity to model accelerated changes at the human tissue level in the extreme space environment. Spaceflight-induced muscle atrophy experienced by astronauts shares similar physiological changes to muscle wasting in older adults, known as sarcopenia. These shared attributes provide a rationale for investigating molecular changes in muscle cells exposed to spaceflight that may mimic the underlying pathophysiology of sarcopenia. We report the results from three-dimensional myobundles derived from muscle biopsies from young and older adults, integrated into an autonomous CubeLab™, and flown to the International Space Station (ISS) aboard SpaceX CRS-21 as part of the NIH/NASA funded Tissue Chips in Space program. Global transcriptomic RNA-Seq analyses comparing the myobundles in space and on the ground revealed downregulation of shared transcripts related to myoblast proliferation and muscle differentiation. The analyses also revealed downregulated differentially expressed gene pathways related to muscle metabolism unique to myobundles derived from the older cohort exposed to the space environment compared to ground controls. Gene classes related to inflammatory pathways were downregulated in flight samples cultured from the younger cohort compared to ground controls. Our muscle tissue chip platform provides an approach to studying the cell autonomous effects of spaceflight on muscle cell biology that may not be appreciated on the whole organ or organism level and sets the stage for continued data collection from muscle tissue chip experimentation in microgravity. We also report on the challenges and opportunities for conducting autonomous tissue-on-chip CubeLabTM payloads on the ISS.

Microphysiological system for studying contractile differences in young, active, and old, sedentary adult derived skeletal muscle cells.

Microphysiological systems (MPS), also referred to as tissue chips, incorporating 3D skeletal myobundles are a novel approach for physiological and pharmacological studies to uncover new medical treatments for sarcopenia. We characterize a MPS in which engineered skeletal muscle myobundles derived from donor-specific satellite cells that model aged phenotypes are encapsulated in a perfused tissue chip platform containing platinum electrodes. Our myobundles were derived from CD56+ myogenic cells obtained via percutaneous biopsy of the vastus lateralis from adults phenotyped by age and physical activity. Following 17 days differentiation including 5 days of a 3 V, 2 Hz electrical stimulation regime, the myobundles exhibited fused myotube alignment and upregulation of myogenic, myofiber assembly, signaling and contractile genes as demonstrated by gene array profiling and localization of key components of the sarcomere. Our results demonstrate that myobundles derived from the young, active (YA) group showed high intensity immunofluorescent staining of α-actinin proteins and responded to electrical stimuli with a ~1 µm displacement magnitude compared with non-stimulated myobundles. Myobundles derived from older sedentary group (OS) did not display a synchronous contraction response. Hypertrophic potential is increased in YA-derived myobundles in response to stimulation as shown by upregulation of insulin growth factor (IGF-1), α-actinin (ACTN3, ACTA1) and fast twitch troponin protein (TNNI2) compared with OS-derived myobundles. Our MPS mimics disease states of muscle decline and thus provides an aged system and experimental platform to investigate electrical stimulation mimicking exercise regimes and may be adapted to long duration studies of compound efficacy and toxicity for therapeutic evaluation against sarcopenia.

Resorbable plating system stabilizes tissue-engineered intervertebral discs implanted ex vivo in canine cervical spines.

Total disc replacement using tissue-engineered intervertebral discs (TE-IVDs) may offer a biological alternative to treat radiculopathy caused by disc degeneration. A composite TE-IVD was previously developed and evaluated in rat tail and beagle cervical spine models in vivo. Although cell viability and tissue integration into host tissue were promising, significant implant displacement occurred at multiple spinal levels. The goal of the present study was to assess the effects of a resorbable plating system on the stiffness of motion segments and stability of tissue-engineered implants subjected to axial compression. Canine motion segments from levels C2/C3 to C5/C6 were assessed as intact (CTRL), after discectomy (Dx), with an implanted TE-IVD only (PLATE-), and with a TE-IVD combined with an attached resorbable plate (PLATE+). Segments under PLATE+ conditions fully restored separation between endplates and showed significantly higher compressive stiffness than segments under PLATE- conditions. Plated segments partially restored more than 25% of the CTRL motion segment stiffness. Plate attachment also prevented implant extrusion from the disc space at 50% compressive strain, and this effect was more significant in segments from levels C3/C4 when compared to segments from level C5/C6. These results suggest that stabilization of motion segments via resorbable plating assists TE-IVD retention in the disc space while allowing the opportunity for implants to fully integrate into the host tissue and achieve optimal restoration of spine biomechanics.