Behaviour Of HBMSC Affected By Electrospun Fibrous Membranes Containing Nanoparticles Encapsulated With MGF

Biological carrier, seed cells and biological factors have become recognized as the threemost important factors in tissue engineering. Our topics used the mechano-growth factorwhich was discovered and named by Yang, generated by cells in response to mechanicalstimulation, then prepared scaffold based on poly caprolactone containing dextrannanospheres encapsulated with MGF using the technology of electrospinning and phaseseparation. Then, from the difference in the biological behavior of hBMSC to study theprotective effect of dextran microspheres on MGF biological activity. This article containsdetailed contents as follows:①The isolation, cultivation and identification of primary human mesenchymal stemcellsIsolating and culturing hBMSC by density gradient centrifugation, then observed the cellmorphology and proliferation rate. CD14, CD34, CD73, CD90and CD105antibodies wereused to stained, following to detect cell cycle status by flow cytometry to determine the purityof the cells. Last，we used the induction medium to induce cell differentiated into fat cell andosteogenic cell to study the multipotential differentiation of hBMSC which laying thefoundation for the next experiment.②Preparation and characterization ofnanoparticlesencapsulatedwithMGFWe prepared empty microspheres and MGF-Loaded dextran glassy nanoparticles（MGF-DGNs）for the next electrospun nanofiber though the method of phase separation,then observed the shape of nanospheres by field emission scanning electron microscopy（FETEM）.③Preparation and characterization of electrospun scaffoldsUsing electrospinning technique to prepare four fibrous scaffolds, which were pure PCLsolution (P); admixturing PCL solution and200μg MGF solution(PM); blending PCLsolution and100mg empty dextran microspheres(PD); mixing PCL solution and100mgdextran microspheres encapsulated with200μg MGF(PDM). Then the nanofibers werecharacterized for various physicochemical properties, such as FESEM, Transmission electronmicroscopy（TEM）, Fourier-transform infrared spectroscopy(FTIR) and ASAP2020analyzer.④ThebiologicaleffectsofMGF on hBMSC behaviorP, PM, PD, PDM scaffolds were planted with hBMSC, using spectrophotometric microplate reader and confocal laser scanning microscopy(CLSM) to observe theproliferation and migration of hBMSC, detecting the impact of the release MGF frommaterial on hBMSC biological behavior.Real-time PCR detected MGF is highly expressed in hBMSC seeded on PDM. Theenhanced cell proliferation and migration show that the MGF-DGNs-loaded PCL fibrousmembrane can release MGF sustainably and secure the bioactivity of MGF better than otherPCL fibrous membranes, which may help find MGF-based fibrous membranes application ofMSC in tissue engineering.