Co-delivery of oxytocin, a p38α/β inhibitor, and fibronectin within a soft hydrogel niche synergistically reactivates...

IF a single intramuscular injection of oxytocin (1 µg/kg, local depot) co-formulated with the p38α/β MAPK inhibitor SB202190 (10 µM final tissue concentration) within a fibronectin-functionalized soft polyethylene glycol (PEG) hydrogel (~2 kPa stiffness, mimicking young basal lamina mechanics) is administered to the tibialis anterior of aged (24-month) male C57BL/6 mice 24 hours prior to cardiotoxin-induced injury,

THEN a significant restoration of satellite cell (SC) proliferative output — measured as ≥2-fold increase in EdU+ Pax7+ cells at 72 hours post-injury versus aged vehicle controls — will be observed alongside a ≥50% reduction in the proportion of p16INK4a-high geroconverted SCs, improved myofiber cross-sectional area at 21 days, and reduced collagen I deposition, all compared to aged mice receiving oxytocin alone or p38i alone,

BECAUSE the following mechanistic chain is activated:

1. Ligand restoration step: Circulating oxytocin declines with age, but OXTR remains expressed on aged satellite cells; exogenous OT binding to OXTR activates ERK1/2 phosphorylation, which is the requisite signal for aged SCs to exit G0 quiescence and enter the cell cycle. (Oxytocin administration rescues proliferation and differentiation deficits and improves regeneration in aged mice, in part via MAPK/ERK activation)[https://doi.org/10.1038/nrm.2016.7]

2. Geroconversion vulnerability step: However, when aged satellite cells are driven into proliferation in isolation, they are acutely vulnerable to geroconversion: the chronic elevation of p38α/β MAPK in aged SCs couples ERK-driven proliferative pressure to upregulation of p16INK4a, repression of pRb, and entry into irreversible senescence rather than productive myogenesis. (Geriatric satellite cells expressing high levels of p16INK4a under proliferative pressure exhibited reduced phosphorylated pRb levels)[https://doi.org/10.4161/15384101.2014.965072]

3. p38 inhibition blocks geroconversion and restores self-renewal: Co-delivery of SB202190 at the site of OT-mediated SC activation suppresses chronic p38α/β activity, preventing the p16INK4a-driven geroconversion that would otherwise consume newly activated aged SCs; p38 inhibition alone has demonstrated that it restores the self-renewal division bias in aged SCs. (p38α/β signaling has context-dependent roles in myogenesis: it suppresses expression of stem cell self-renewal genes in aged muscle)[https://doi.org/10.1038/nm.3464]

4. Biomechanical niche rescue step: The aged ECM undergoes fibronectin (FN) loss, which impairs integrin-mediated FAK signaling critical for SC quiescence maintenance and activation competence; delivering the intervention within a FN-functionalized soft hydrogel (~2 kPa) reconstitutes this lost ECM signal. (Loss of fibronectin from the regenerative niche is a critical mechanism underlying MuSC aging; reduced FN diminishes integrin-mediated FAK signaling)[https://doi.org/10.1038/nm.4126]

5. **Biophysical-pharmacolog...

SENS category: GlycoSENS

Key references: • doi.org/10.1038/nrm.2016.7] • doi.org/10.4161/15384101.2014.965072] • doi.org/10.1038/nm.3464] • doi.org/10.1038/nm.4126] • doi.org/10.3389/fgene.2015.00059].