Prostaglandin E2 produced by alginate-encapsulated mesenchymal stromal cells modulates the astrocyte inflammatory response

Astroglia are well known for their role in propagating secondary injury following brain trauma. Modulation of this injury cascade, including inflammation, is essential to repair and recovery. Mesenchymal stromal cells (MSCs) have been demonstrated as trophic mediators in several models of secondary CNS injury, however, there has been varied success with the use of direct implantation due to a failure to persist at the injury site. To achieve sustained therapeutic benefit, we have encapsulated MSCs in alginate microspheres and evaluated the ability of these encapsulated MSCs to attenuate neuro-inflammation. In this study, astroglial cultures were administered lipopolysaccharide (LPS) to induce inflammation and immediately co-cultured with encapsulated or monolayer human MSCs. Cultures were assayed for the pro-inflammatory cytokine tumor necrosis factor alpha (TNF-[Formula: see text] produced by astroglia, MSC-produced prostaglandin E2, and expression of neurotrophin-associated genes. We found that encapsulated MSCs significantly reduced TNF-[Formula: see text] produced by LPS-stimulated astrocytes more effectively than monolayer MSCs, and this enhanced benefit commences earlier than that of monolayer MSCs. Furthermore, in support of previous findings, encapsulated MSCs constitutively produced high levels of PGE2, while monolayer MSCs required the presence of inflammatory stimuli to induce PGE2production. The early, constitutive presence of PGE2significantly reduced astrocyte-produced TNF-[Formula: see text], while delayed administration had no effect. Finally, MSC-produced PGE2was not only capable of modulating inflammation, but appeared to have an additional role in stimulating astrocyte neurotrophin production. Overall, these results support the enhanced benefit of encapsulated MSC treatment, both in modulating the inflammatory response and providing neuroprotection.</jats:p

A 3D Tissue Model of Traumatic Brain Injury with Excitotoxicity That Is Inhibited by Chronic Exposure to Gabapentinoids

Injury progression associated with cerebral laceration is insidious. Following the initial trauma, brain tissues become hyperexcitable, begetting further damage that compounds the initial impact over time. Clinicians have adopted several strategies to mitigate the effects of secondary brain injury; however, higher throughput screening tools with modular flexibility are needed to expedite mechanistic studies and drug discovery that will contribute to the enhanced protection, repair, and even the regeneration of neural tissues. Here we present a novel bioengineered cortical brain model of traumatic brain injury (TBI) that displays characteristics of primary and secondary injury, including an outwardly radiating cell death phenotype and increased glutamate release with excitotoxic features. DNA content and tissue function were normalized by high-concentration, chronic administrations of gabapentinoids. Additional experiments suggested that the treatment effects were likely neuroprotective rather than regenerative, as evidenced by the drug-mediated decreases in cell excitability and an absence of drug-induced proliferation. We conclude that the present model of traumatic brain injury demonstrates validity and can serve as a customizable experimental platform to assess the individual contribution of cell types on TBI progression, as well as to screen anti-excitotoxic and pro-regenerative compounds

Evaluation of the Osteoinductive Capacity of Polydopamine-Coated Poly(ε-caprolactone) Diacrylate Shape Memory Foams

Recently, a novel shape memory polymer foam based on the photopolymerization of poly­(ε-caprolactone) diacrylate (PCLDA) has been developed. These PCLDA foams enter a temporary softened state when briefly treated with warm saline (<i>T</i>_saline > <i>T</i>_m of PCLDA), allowing them to conform to irregular bone defect “boundaries” prior to shape setting. When coated with a mechanically stable polydopamine (PD) layer, these PCLDA foams have previously been demonstrated to induce hydroxyapatite deposition. In the present study, the osteoinductivity of these “self-fitting” PD-coated PCLDA (PD–PCLDA) materials was evaluated relative to uncoated PCLDA (U-PCLDA) controls using bone marrow-derived human mesenchymal stem cells (h-MSCs). When cultured in the absence of osteogenic media supplements, PD–PCLDA scaffolds expressed similar levels of Runx2, alkaline phosphatase, and osteopontin protein as U-PCLDA scaffolds cultured in the presence of osteogenic media supplements. In addition, PD–PCLDA scaffolds cultured without osteogenic supplements did not significantly promote undesired lineage progression (e.g., adipogenesis or chondrogenesis) of h-MSCs. Cumulatively, these data indicate that PD–PCLDA materials display increased osteoinductivity relative to U-PCLDA substrates. Future studies will examine tethered osteogenic factors or peptides toward augmenting the osteoinductive properties of the PD–PCLDA foams