The Fabrication Of An ICA-SF/PLCL Nanofibrous Membrane By Coaxial Electrospinning And Its Effect On Bone Regeneration In Vitro

Objective: In this study,we fabricated an ICA-SF/PLCL nanofibrous membrane by coaxial electrospinning.The physical and chemical properties,mechanical properties,in vitro release and degradation ability were evaluated systematically.Cell seeding experiments were performed to evaluate the biocompatibility and osteoblastic ability of the membrane in vitro.To explore the possibility of the nanofibrous membrane as a GBR barrier membrane for promoting bone regeneration.Methods:(1)The parameters during the coaxial electrospinning process were confirmed from our previous work : the voltage was 15 kV,the receiving distance was 15 cm,the advancing velocities of the core layer and shell solution were 0.1 mL·h-1 and 1.0 mL·h-1,respectively,.the relative humidity was 65% and the temperature was 25 ?.10-5 mol·L-1 ICA solution was used as the core layer solution and 8% SF/PLCL solution(m/m=30:70)was used as the shell layer solution,the ICA-SF/PLCL nanofibrous membrane was fabricated by coaxial electrospinning.(2)The surface morphology of the membrane was observed by scanning electron microscopy(SEM)and the structure of the composite nanofiber was observed by transmission electron microscopy(TEM);The hydrophobicity of the membrane was tested by contact angle test;Using universal mechanical testing machine to test the mechanical properties;To detect the ICA released from the membrane,a UV-Vis was used;The morphological changes of the membranes under the action of the degradation medium were observed by SEM and the mechanical properties changes were studied by the universal testing machine.(3)In the bone marrow mesenchymal stem cells(BMSCs)seeded experiments,the cells adhesion and proliferation were observed by SEM and MTT assay.Furthermore,alizarin red staining and ALP activity assay were used to evaluate the ability of the membrane to promote bone formation.Results:(1)An ICA-SF/PLCL nanofibrous membrane was fabricated by coaxial electrospinning technology.(2)SEM showed the morphology of the membrane surface was good,the diameter and distribution of the fibers were uniform,which showed a three-dimensional network structure.TEM observed that there was a core-shell structure along the long axis of the fiber.The hydrophilic angle of the membrane was about 125.72±4.93 °,which was lower than that of pure PLCL spinning membrane,the hydrophilicity of the membrane was improved.The mechanical property tests showed the tensile strength of the membrane was about 5.54±0.46 MPa,which could meet the requirement of clinical GBR barrier membrane strength.The drug-release study showed that ICA released from the membrane could be divided into the initial short release phase(0 ~ 5 d,the cumulative release rate was 41.06±0.06%)and the slow and sustained-release stage(6 ~ 30 d,the final cumulative release rate reaching 82.09±1.86%),the drug release time was effectively prolonged.In vitro,degradation experiments showed that the surface of the membrane gradually became rough,and even some of the sizes of hole,some of these fibers fracture,thus,the tensile strength of the membrane was gradually reduced.(3)The cells seeding experiments showed that BMSCs adhered on the membrane surface and grew well.Alizarin red staining showed obvious calcified nodule formation,and intracellular ALP activity was increased significantly.Conclusion: An ICA-SF/PLCL nanofibrous membrane was successfully fabricated by coaxial electrospinning technology.Its surface morphology was good,the hydrophilicity and mechanical property were excellent,had the ability of biodegradability and drug release effect,excellent biocompatibility,especially had the ability to promote bone formation in vitro,so this kind of drug-loaded nanofiber membrane prepared by the combination of GBR and growth factor and drug release technology is expected to be a new and effective GBR barrier membrane for bone regeneration..