Evaluating the protective effects of dexamethasone and electrospun mesh combination on primary human mixed retinal cells under hyperglycemic stress.

Chronic inflammation is a leading cause of neurodegeneration and vision loss in hyperglycemia-associated conditions such as diabetic retinopathy. Corticosteroid injections are widely used for treatment but suffer from limitations such as rapid drug clearance, short drug half-lives and frequent administration. While drug release from biomaterial carriers can overcome these shortcomings, evaluating the combined effects of corticosteroids and polymeric matrices under hyperglycemic stress is an important step towards aiding translation. In this study, we investigated the effects of dexamethasone (DEX) and electrospun mesh combination on primary human mixed retinal cells under normal and hyperglycemic culture conditions. DEX-incorporated poly(lactide-co-glycolide) (PLGA) meshes were prepared and characterized for architecture, chemistry, drug distribution and in vitro release. The meshes exhibited cumulative in vitro drug release of 39.5 % over 2 months at a near constant rate. Under normal culture conditions, DEX-PLGA meshes promoted significantly higher viability of mixed retinal cells than the control groups but without adverse phenotypic activation. Under hyperglycemic conditions, DEX supplementation resulted in higher viability than the control, although the highest viability was achieved only when DEX was added to cells cultured on PLGA fibers. The combination of DEX and PLGA fibers also promoted higher mRNA expression of the antioxidant GSH under hyperglycemia. Importantly, the largest reduction in the production of pro-inflammatory cytokines viz., MMP-9, IL-6, IL-8 and VEGF-R1 was observed for the DEX and PLGA combination. Our study reveals a combined effect of DEX and electrospun fibers in combating hyperglycemia-driven pro-inflammatory responses, which can aid the development of DEX-loaded electrospun implants for diabetes-driven retinal conditions.

Electrospun fiber-based strategies for controlling early innate immune cell responses: Towards immunomodulatory mesh designs that facilitate robust tissue repair.

Electrospun fibrous meshes are widely used for tissue repair due to their ability to guide a host of cell responses including phenotypic differentiation and tissue maturation. A critical factor determining the eventual biological outcomes of mesh-based regeneration strategies is the early innate immune response following implantation. The natural healing process involves a sequence of tightly regulated, temporally varying and delicately balanced pro-/anti-inflammatory events which together promote mesh integration with host tissue. Matrix designs that do not account for the immune milieu can result in dysregulation, chronic inflammation and fibrous capsule formation, thus obliterating potential therapeutic outcomes. In this review, we provide systematic insights into the effects of specific fiber/mesh properties and mechanical stimulation on the responses of early innate immune modulators viz., neutrophils, monocytes and macrophages. We identify matrix characteristics that promote anti-inflammatory immune phenotypes, and we correlate such responses with pro-regenerative in vivo outcomes. We also discuss recent advances in 3D fabrication technologies, bioactive functionalization approaches and biomimetic/bioinspired immunomodulatory mesh design strategies for tissue repair and wound healing. The mechanobiological insights and immunoregulatory strategies discussed herein can help improve the translational outcomes of fiber-based regeneration. STATEMENT OF SIGNIFICANCE: The crucial role played by immune cells in promoting biomaterial-based tissue regeneration is being increasingly recognized. In this review focusing on the interactions of innate immune cells with electrospun fibrous meshes, we systematically elucidate the effects of the fiber microenvironment and mechanical stimulation on biological responses, and build upon these insights to inform the rational design of immunomodulatory meshes for effective tissue repair. We discuss state-of-the-art fabrication methods and mechanobiological advances that permit the orchestration of temporally controlled phenotypic switches in immune cells during different phases of healing. The design strategies discussed herein can also be leveraged to target several complex autoimmune and inflammatory diseases.

Electrospun meshes intrinsically promote M2 polarization of microglia under hypoxia and offer protection from hypoxia-driven cell death.

In this study, we offer new insights into the contrasting effects of electrospun fiber orientation on microglial polarization under normoxia and hypoxia, and establish for the first time, the intrinsically protective roles of electrospun meshes against hypoxia-induced microglial responses. First, resting microglia were cultured under normoxia on poly(caprolactone) fibers possessing two distinctly different fiber orientations. Matrix-guided differences in cell shape/orientation and differentially expressed Rho GTPases (RhoA, Rac1, Cdc42) were well-correlated with the randomly oriented fibers inducing a pro-inflammatory phenotype and the aligned fibers sustaining a resting phenotype. Upon subsequent hypoxia induction, both sets of meshes offered protection from hypoxia-induced damage by promoting a radical phenotypic switch and beneficially altering the M2/M1 ratio to different extents. Compared to 2D hypoxic controls, meshes significantly suppressed the expression of pro-inflammatory markers (IL-6, TNF-α) and induced drastically higher expression of anti-inflammatory (IL-4, IL-10, VEGF-189) and neuroprotective (Nrf-2) markers. Consistent with this M2 polarization, the expression of Rho GTPases was significantly lower in the mesh groups under hypoxia compared to normoxic culture. Moreover, meshes-particularly with aligned fibers-promoted higher cell viability, suppressed caspase 3/8 and LC-3 expression and promoted LAMP-1 and LAMP-2 expression, which suggested the mitigation of apoptotic/autophagic cell death via a lysosomal membrane-stabilization mechanism. Notably, all protective effects under hypoxia were observed in the absence of additional soluble cues. Our results offer promise for leveraging the intrinsic therapeutic potential of electrospun meshes in degenerative diseases where microglial dysfunction, hypoxia and inflammation are implicated.