Reactive oxygen-scavenging polydopamine nanoparticle coated 3D nanofibrous scaffolds for improved osteogenesis: Toward an aging in vivo bone regeneration model.

Tissue engineered scaffolds aimed at the repair of critical-sized bone defects lack adequate consideration for our aging society. Establishing an effective aged in vitro model that translates to animals is a significant unmet challenge. The in vivo aged environment is complex and highly nuanced, making it difficult to model in the context of bone repair. In this work, 3D nanofibrous scaffolds generated by the thermally-induced self-agglomeration (TISA) technique were functionalized with polydopamine nanoparticles (PD NPs) as a tool to improve drug binding capacity and scavenge reactive oxygen species (ROS), an excessive build-up that dampens the healing process in aged tissues. PD NPs were reduced by ascorbic acid (rPD) to further improve hydrogen peroxide (H2O2) scavenging capabilities, where we hypothesized that these functionalized scaffolds could rescue ROS-affected osteoblastic differentiation in vitro and improve new bone formation in an aged mouse model. rPDs demonstrated improved H2O2 scavenging activity compared to neat PD NPs, although both NP groups rescued the alkaline phosphatase activity (ALP) of MC3T3-E1 cells in presence of H2O2. Additionally, BMP2-induced osteogenic differentiation, both ALP and mineralization, was significantly improved in the presence of PD or rPD NPs on TISA scaffolds. While in vitro data showed favorable results aimed at improving osteogenic differentiation by PD or rPD NPs, in vivo studies did not note similar improvements in ectopic bone formation an aged model, suggesting that further nuance in material design is required to effectively translate to improved in vivo results in aged animal models.

An Elastic Mineralized 3D Electrospun PCL Nanofibrous Scaffold for Drug Release and Bone Tissue Engineering.

Complex shaped and critical-sized bone defects have been a clinical challenge for many years. Scaffold-based strategies such as hydrogels provide localized drug release while filling complex defect shapes, but ultimately possess weaknesses in low mechanical strength alongside a lack of macroporous and collagen-mimicking nanofibrous structures. Thus, there is a demand for mechanically strong, extracellular matrix (ECM) mimicking scaffolds that can robustly fit complex shaped critical sized defects and simultaneously provide localized, sustained, multiple growth factor release. We therefore developed a composite, bi-phasic PCL/hydroxyapatite (HA) 3D nanofibrous (NF) scaffold for bone tissue regeneration by using our innovative electrospun-based thermally induced self-agglomeration (TISA) technique. One intriguing feature of our ECM-mimicking TISA scaffolds is that they are highly elastic and porous even after evenly coated with minerals and can easily be pressed to fit different defect shapes. Furthermore, the bio-mimetic mineral deposition technique allowed us to simultaneously encapsulate different type of drugs, e.g., proteins and small molecules, on TISA scaffolds under physiologically mild conditions. Compared to scaffolds with physically surface-adsorbed phenamil, a BMP2 signaling agonist, incorporated phenamil composite scaffolds indicated less burst release and longer lasting sustained release of phenamil with subsequently improved osteogenic differentiation of cells in vitro. Overall, our study indicated that the innovative press-fit 3D NF composite scaffold may be a robust tool for multiple-drug delivery and bone tissue engineering.

Local application of MDL28170-loaded PCL film improves functional recovery by preserving survival of motor neurons after traumatic spinal cord injury.

Neuronal death and organization degeneration can happen inordinately after spinal cord injury (SCI), which lead to nerve dysfunction. We aimed to determine whether local application of a cell permeable calpain I inhibitor (MDL28170) can promote SCI recovery by increasing neuronal cell viability. MDL28170-loaded polycaprolactone (PCL) film was fabricated. Scanning electron microscopy showed the surface of PCL film was smooth with holes (diameter at µM level). The PCL film was non-toxic, biological compatibility, and had good neuron adhension and slow release characteristic. MDL28170 increased VSC4.1 motor neurons' viability under tunicamycin (an endoplasmic reticulum stress) induced injury. In a traumatic SCI rat model, MDL28170-loaded PCL film reduced the area of lesion cavity, and promoted recovery of locomotor behavior. Moreover, the expression of GAP-43 was upregulated after MDL28170-loaded PCL film treatment. Thus, our findings demonstrated that localized delivery of MDL28170 could promote SCI recovery by inhibiting endoplasmic reticulum stress, preserving survival of the motor neurons, which may point out a promising therapeutic target for treating SCI patient.

[Effect of nano-liposome sustained elemene in inducing cell apoptosis of C6 glioma].

OBJECTIVE: To study the effect of nano-liposome sustained elemene in inducing cell apoptosis of C6 glioma and to explore its influence on the expression of caspase-3 gene. METHODS: C6 glioma cells were cultured in medium with the same amount of nano-liposome sustained elemene and common elemene respectively, also in blank medium for control. The status of cell apoptosis was determined by flow cytometry at 0, 48 h and 72 h, and the expression of Caspase-3 protein was measured simultaneously by immunohistochemistry assay. RESULTS: Marked apoptosis presented in cells cultured in the medium with nano-liposome sustained elemene or common elemene at 48 h and 72 h, with the apoptotic rate significantly higher than that in the control. At the same time, Caspase-3 protein expression raised significantly in cells cultured in medium with either kinds of elemene, showing significant difference when compared with that in the control. CONCLUSION: Elemene has significant apoptosis promoting and Caspase-3 protein expression inducing effect on C6 glioma cells, which could be facilitated by nano-liposome bearing.

Electrospinning of poly(glycerol sebacate)-based nanofibers for nerve tissue engineering.

Nerve tissue engineering (TE) requires biomimetic scaffolds providing essential chemical and topographical cues for nerve regeneration. Poly(glycerol sebacate) (PGS) is a biodegradable and elastic polymer that has gained great interest as a TE scaffolding biomaterial. However, uncured PGS is difficult to be electrospun into nanofibers. PGS would, therefore, require the addition of electrospinning agents. In this study, we modified PGS by using atom transfer radical polymerization (ATRP) to synthesize PGS-based copolymers with methyl methacrylate (MMA). The synthesized PGS-PMMA copolymer showed a molecular weight of 82kDa and a glass transition temperature of 115°C. More importantly, the PGS-PMMA could be easily electrospun into nanofiber with a fiber diameter of 167±33nm. Blending gelatin into PGS-PMMA nanofibers was found to increase its hydrophilicity and biocompatibility. Rat PC12 cells were seeded onto the PGS-PMMA/gelatin nanofibers to investigate their potential for nerve regeneration. It was found that gelatin-containing PGS-based nanofibers promoted cell proliferation. The elongated cell morphology observed on such nanofibers indicated that the scaffolds could induce the neurite outgrowth of the nerve stem cells. Overall, our study suggested that the synthesis of PGS-based copolymers might be a promising approach to enhance their processability, and therefore advancing bioscaffold engineering for various TE applications.

Synergistic effect of topography, surface chemistry and conductivity of the electrospun nanofibrous scaffold on cellular response of PC12 cells.

Electrospun nanofibrous nerve implants is a promising therapy for peripheral nerve injury, and its performance can be tailored by chemical cues, topographical features as well as electrical properties. In this paper, a surface modified, electrically conductive, aligned nanofibrous scaffold composed of poly (lactic acid) (PLA) and polypyrrole (Ppy), referred to as o-PLAPpy_A, was fabricated for nerve regeneration. The morphology, surface chemistry and hydrophilicity of nanofibers were characterized by Scanning Electron Microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and water contact angle, respectively. The effects of these nanofibers on neuronal differentiation using PC12 cells were evaluated. A hydrophilic surface was created by Poly-ornithine coating, which was able to provide a better environment for cell attachment, and furthermore aligned fibers were proved to be able to guide PC12 cells grow along the fiber direction and be beneficial for neurite outgrowth. The cellular response of PC12 cells to pulsed electrical stimulation was evaluated by NF 200 and alpha tubulin expression, indicating that electrical stimulation with a voltage of 40mV could enhance the neurite outgrowth. The PC12 cells stimulated with electrical shock showed greater level of neurite outgrowth and smaller cell body size. Moreover, the PC12 cells under electrical stimulation showed better viability. In summary, the o-PLAPpy_A nanofibrous scaffold supported the attachment, proliferation and differentiation of PC12 cells in the absence of electrical stimulation, which could be potential candidate for nerve regeneration applications.

Emulsion electrospinning of polycaprolactone: influence of surfactant type towards the scaffold properties.

Producing uniform nanofibers in high quality by electrospinning remains a huge challenge, especially using low concentrated polymer solutions. However, emulsion electrospinning assists to produce nanofibers from less concentrated polymer solutions compared to the traditional electrospinning process. The influence of individual surfactants towards the morphology of the emulsion electrospun poly (ɛ-caprolactone)/bovine serum albumin (PCL/BSA) nanofibers were investigated by using (i) non-ionic surfactant sorbitane monooleate (Span80); (ii) anionic sodium dodecyl sulfate (SDS); and (iii) cationic benzyltriethylammonium chloride, and poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer Pluronic F108 of different concentrations. The morphology, along with the chemical and mechanical properties of the fibers, was evaluated by field emission scanning electron microscopy, attenuated total reflectance Fourier transform infrared spectroscopy, differential scanning calorimetry, water contact angle, and tensile tester. With the addition of surfactants, the electrospinnability of dilute PCL solution was enhanced, with either branched or uniform fibers were obtained. Electrospinning of an emulsion containing 0.4% (w/v) SDS produced the smallest and the most uniform nanofibers (167 ± 39 nm), which was attributed to the high conductivity of the solution. Analysis revealed that the emulsion electrospun nanofibers containing different surfactants and surfactant concentrations differ in fiber morphology and mechanical properties. Results suggest that surfactants have the ability to modulate the fiber morphology via electrostatic and hydrogen bonding, depending on their chemical structure.