ABSTRACT
Diamond-like carbon (DLC) was modified using a UV functionalization method to introduce surface-bound amine and aldehyde groups. The functionalization process rendered the DLC more hydrophilic and significantly increased the viability of neurons seeded to the surface. The amine functionalized DLC promoted adhesion of neurons and fostered neurite outgrowth to a degree indistinguishable from positive control substrates (glass coated with poly-L-lysine). The aldehyde-functionalized surfaces performed comparably to the amine functionalized surfaces and both additionally supported the adhesion and growth of primary rat Schwann cells. DLC has many properties that are desirable in biomaterials. With the UV functionalization method demonstrated here it may be possible to harness these properties for the development of implantable devices to interface with the nervous system.
Subject(s)
Biocompatible Materials/chemistry , Diamond/chemistry , Schwann Cells/drug effects , Aldehydes/chemistry , Amines/chemistry , Animals , Biocompatible Materials/toxicity , Cell Differentiation/drug effects , Cell Line , Cell Survival/drug effects , Cells, Cultured , Diamond/toxicity , Male , Mice , Neural Prostheses , Photochemical Processes , Prosthesis Design , Rats , Rats, WistarABSTRACT
In this study, we report the production of amine functionalized nanodiamond. The amine functionalized nanodiamond forms a conformal monolayer on a negatively charged surface produced via plasma polymerization of acrylic acid. Nanodiamond terminated surfaces were studied as substrates for neuronal cell culture. NG108-15 neuroblastoma-glioma hybrid cells were successfully cultured upon amine functionalized nanodiamond coated surfaces for between 1 and 7 d. Additionally, primary dorsal root ganglion (DRG) neurons and Schwann cells isolated from Wistar rats were also successfully cultured over a period of 21 d illustrating the potential of the coating for applications in the treatment of peripheral nerve injury.
Subject(s)
Amines/chemistry , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Nanodiamonds/chemistry , Neurites/drug effects , Actins/chemistry , Animals , Fluoroacetates/chemistry , Ganglia, Spinal/metabolism , Glioma/drug therapy , Humans , Male , Nanotechnology , Neurites/metabolism , Neuroblastoma/drug therapy , Neurons/metabolism , Photoelectron Spectroscopy , Rats , Rats, Wistar , Surface Properties , Trifluoroacetic Acid/chemistryABSTRACT
In this study, we explore the production of well-defined macroscopic scaffolds with two-photon polymerization (2PP) and their use as neural tissue engineering scaffolds. We also demonstrate that these 3D scaffolds can be replicated via soft lithography, which increases production efficiency. Photopolymerizable polylactic acid (PLA) was used to produce scaffolds by 2PP and soft lithography. We assessed the biocompatibility of these scaffolds using an SH-SY5Y human neuronal cell line and primary cultured rat Schwann cells (of direct relevance to the repair of nerve injuries). A Comet assay with SH-SY5Y human neuronal cells revealed minimal DNA damage after washing the photocured material for 7 days in ethanol. Additionally, thin films and 3D scaffolds of the photocured PLA sustained a high degree of Schwann cell purity (99%), enabled proliferation over 7 days and provided a suitable substrate for supporting Schwann cell adhesion such that bi-polar and tri-polar morphologies were observed. Evidence of orthogonally aligned and organized actin thin filaments and the formation of focal contacts were observed for the majority of Schwann cells. In summary, this work supports the use of PLA as a suitable material for supporting Schwann cell growth and in turn use of 3D soft lithography for the synthesis of neural scaffolds in nerve repair.
Subject(s)
Polymerization/radiation effects , Schwann Cells/cytology , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Biotechnology/instrumentation , Biotechnology/methods , Cell Growth Processes/physiology , Cell Line , Cell Survival/physiology , Cells, Cultured , Immunohistochemistry , Lactic Acid/chemistry , Lactic Acid/radiation effects , Male , Microscopy, Confocal , Microscopy, Electron, Scanning , Microtechnology/instrumentation , Microtechnology/methods , Photochemical Processes , Polyesters , Polymers/chemistry , Polymers/radiation effects , Rats , Rats, WistarABSTRACT
This study reports on the production of high-resolution 3D structures of polylactide-based materials via multi-photon polymerization and explores their use as neural tissue engineering scaffolds. To achieve this, a liquid polylactide resin was synthesized in house and rendered photocurable via attaching methacrylate groups to the hydroxyl end groups of the small molecular weight prepolymer. This resin cures easily under UV irradiation, using a mercury lamp, and under femtosecond IR irradiation. The results showed that the photocurable polylactide (PLA) resin can be readily structured via direct laser write (DLW) with a femtosecond Ti:sapphire laser and submicrometer structures can be produced. The maximum resolution achieved is 800 nm. Neuroblastoma cells were grown on thin films of the cured PLA material, and cell viability and proliferation assays revealed good biocompatibility of the material. Additionally, PC12 and NG108-15 neuroblastoma growth on bespoke scaffolds was studied in more detail to assess potential applications for neuronal implants of this material.
Subject(s)
Lasers , Nerve Tissue/physiology , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Cell Proliferation/drug effects , Microscopy, Fluorescence , Nerve Tissue/cytology , Nerve Tissue/drug effects , Nerve Tissue/ultrastructure , Neurons/cytology , Neurons/drug effects , Neurons/ultrastructure , PC12 Cells , Polyesters/chemical synthesis , Polyesters/chemistry , Polyesters/pharmacology , Rats , Surface Properties/drug effectsABSTRACT
This paper reports a two-year study on the methods used to evaluate 63 junior occupational therapy students in a musculo-skeletal anatomy course offered at the University of Florida. Laboratory learning processes were not significantly different compared to class learning processes as shown by test results. There was also no significant difference between laboratory and written examination scores in anatomy when compared with final grades in subsequent courses with both laboratory and lecture components. This study was undertaken because of the varying attitudes of students and faculty and their knowledge about the most efficient and productive manner in which to conduct the course. Very little research on educational methods has been completed in our profession and the validation of current methods is needed.
