Manipulation of Oxygen Tension in Damaged Regions via Hypoxia-Induced IPN Hydrogel Microspheres for Intervertebral Disc Regeneration

Disruption of low oxygen tension homeostasis during intervertebral disc degeneration inhibits endogenous stem cell viability and function, posing a challenge for endogenous regeneration. Here, to achieve sustained hypoxia manipulation, constructed hypoxia-inducible interpenetrating polymer network (IPN) hydrogel microspheres (HIMS) are constructed by microfluidics to integrate the hypoxic system with a stabilizing network. The IPN is synthesized through a two-step polymerization process, consisting of rapid photo-crosslinked gelatin methacrylate anhydride (GM) polymer I and slow enzyme-crosslinked vanillin-grafted gelatin (GV) polymer II. The enzymatic reaction between GV and laccase is able to create a hypoxic microenvironment to modulate oxygen tension in situ within the injured region. HIMS can reduce microenvironmental oxygen tension by 1/3 and maintain a hypoxic microenvironment for up to 5 days, thereby activating the PI3K/AKT/HIF-1α signaling pathway in endogenous stem cells to promote differentiation into nucleus pulposus-like cells. Additionally, NSC-Exos are loaded onto HIMS to trigger endogenous progenitor/stem cell recruitment and migration. Both in vitro and in vivo assays demonstrate that NSC-Exos@HIMS facilitates stem cell recruitment, targets differentiation, and stimulates extracellular matrix synthesis. Overall, the microspheres established herein provide a novel strategy for manipulating oxygen tension and enhancing endogenous tissue regeneration in injured regions during intervertebral disc degeneration.