Cytocompatibility Research Of Schwann Cells And Neural Stem Cells Co-seeded On Electrospinning Chitosan-PLGA

Objective: The experiment aimed to explore a quick and easy way to acquire quantities of high purity Schwann Cells(SCs) and from the SCs and Neural Stem Cells(NSCs) co-culturing point, to detect the induction of differentiation of SCs’ impact on neural stem cells and the viability and morphology of neurons.Methods: In our study, we composite 0.25% Dispase with 0.2% Collagenase NB4 enzyme digestion for age of 3 to 5 day rat sciatic nerves to the primary generation of SCs. Purified SCs were acquired by Dispase digestion while SCs identification used immunofluorescence staining with S100 specific marker. Differentiation experiments are divided into three groups:SCs group, NSCs group, SCs and NSCs group. Cells were planted respectively in the same differentiation medium condition and after 7 days culturing, GFAP and Map2 specific markers were used to identify differentiated cells( the former was for astrocytes, the latter was for neurons) by method of immunofluorescence staining.Results: Through the method of composite Dispase with Collagenase NB4 enzyme, we can obtain quantities of high purity SCs in a short period. In vitro co-culturing under the same differentiation medium condition,the co-culture group had more neurons, while NSCs group prefer to astrocytes differentiation. Under the condition of differentiation medium, SCs group had no differentiation tendency, cells properties unchanged with S100 specific markers staining positive.Conclusions: Composite enzyme digestion method can obtain numerous high purity SCs quickly; SCs can promote the differentiation of NSCs into neurons under differentiation medium condition.Objective: This study proposed a two-dimensional woven tubular electrostatic spinning braiding with a cylindrical mandrel technique with heat setting coating methods to acquire Chitosan-PLGA nerve conduits with certain elasticity and mechanical strength, in order to better meet the requirements of bridging peripheral nerve defect.Methods: In this study, the use of cylindrical mandrel braiding technique, the inner diameter of the tubular nerve conduits was fixed, which is equal to the metal mandrel diameter. The surface of the braided biological materials nerve conduits were evenly, which is not easy to deform. In this study, we choose the chitosan fiber dissolved in dilute acid solution to attain a mass concentration of 3.5% chitosan slurry. The PLGA biological material nerve conduits were coating with chitosan slurry through heat setting process, so that it has good conformal resistance and excellent mechanical properties, consequently improve its application potentiality. We also placed longitudinal axially arrangement nanofiber filaments inside Chitosan-PLGA to form a three-dimensional structure. The axially nanofibers not only have the role of supporting the tubular structure of nerve conduits, but also simulate the internal environment of regenerated nerves. Phase contrast microscope and scanning electron microscope were performed to observe physical form of the electrostatic spinning Chitosan-PLGA nerve conduits and nanofiber filaments inside the lumen as well as SCs and NSCs morphology on the conduits and nanofibers.Results: Electrostatic spinning Chitosan-PLGA nerve conduit with fixed diameter was uneasy to deform, the nanofibers filaments inside the lumen play a supporting role, and these characteristics make Chitosan-PLGA good conformality and excellent mechanical properties. The Chitosan-PLGA nerve conduits have good adhesion with SCs and NSCs under scanning electron micrope.Conclusion: Electrostatic spinning Chitosan-PLGA nerve conduits with axially nanofiber inside the lumen can simulate the three-dimensional structure for nerve growth, which makes it an ideal scaffold for peripheral nerve injury repair.Objective :To investigate cytocompatibility of the electron spinning chitosan-PLGA nerve conduits with Schwann cells(SCs)and neural stem cells(NSCs) as well as cells adhesion and migration on the arranged nanofibers inside the lumen.Methods:Experiments were divided into three groups. Cells in SCs group, NSCs group, and SCs-NSCs co-culture group were co-seeded with electrostatic spinning Chitosan-PLGA respectively. MTT assay was performed to detect the biocompatibility and survivability of cells in the three groups at 1, 3, 5, 7 days. Phase contrast microscope and scanning electron microscope(SEM) were used to detect cells adhesion and migration on the axially arranged nanofibers inside the lumen.Results: Cell survival rate of the co-culture group is the highest, less in SCs group, NSCs group was the least. SCs and NSCs have good compatibility and adhesion with electrostatic spinning Chitosan-PLGA, and the axial nanofibers in the lumen guide the growth of neurites into a particular direction.Conclusion: Co-culture of SCs and NSCs can facilitate cells compatibility and adhesion with electrostatic spinning Chitosan-PLGA,and the axially arranged nanofibers within the lumen have the effect of directional guide cell growth and migration.