Preparation Of PTH-Fc/SF/PLCL Three-Dimensional Nanofibrous Barrier Membrane By Coaxial Electrospinning And Its Physico-Chemical And Osteogenic Properties

Objective: This experiment intends to use coaxial electrospinning method to prepare PTH-Fc/SF/PLCL three-dimensional nanofiber barrier film,The physicochemical properties of the film were evaluated,the drug release and degradation properties were evaluated in vitro,and the biocompatibility and its ability to promote bone formation in vitro were assessed by cytological experiments.In order to explore the feasibility of the drug-loaded nanofiber barrier membrane as a kind of barrier membrane for promoting bone regeneration.Methods:(1)Based on the previous research of this group,the voltage is 15 kV and the receiving distance is 15 cm,the spinning speeds of the core and shell spinning solutions were 0.1 mL/h and 1.0 mL/h respectively,and the relative ambient humidity was 65%.The ambient temperature was 25°C under the spinning parameters.8% SF/PLCL mixture(30:70 SF/PLCL mass ratio)was used as the shell spinning solution,and PTH-Fc solution was used as the core spinning solution to prepare PTH-Fc./SF/PLCL three-dimensional nanofiber barrier film.(2)Scanning electron microscopy(SEM)was used to observe the surface morphology of the film;Transmission electron microscopy(TEM)was used to further detect the core-shell structure of the composite fiber;The contact angle was used to detect the hydrophilicity and hydrophobicity of the barrier film;The amount of PTH-Fc accumulated in the membrane was measured by UV-visible spectrophotometer,and the drug release performance of the membrane was investigated;The morphology of the membrane surface was measured by SEM and the degradation process was evaluated to evaluate the in vitro degradation capability of the membrane.(3)In vitro experiments,the adhesion and proliferation of bone marrow mesenchymal stem cells(BMSCs)on the membrane surface were observed by scanning electron microscope;Analyze and evaluate the biocompatibility of the membrane by MTT assay;Alkaline phosphatase activity assay and alizarin red staining assay were used to evaluate the ability of the membrane to promote bone formation in vitro.Results:(1)In this experiment,PTH-Fc/SF/PLCL three-dimensional nanofiber barrier films were successfully prepared using coaxial electrospinning technology.(2)It was found by SEM that the surface morphology of the barrier film was excellent,and a three-dimensional three-dimensional network structure was formed by nanofibers with uniform diameter and uniform distribution;The core-shell structure along the long axis of the fiber can be observed by TEM;The contact angle test results show that the contact angle of the barrier film is 125.72 ± 4.73 °,which is smaller than that of pure PLCL spinning film;The drug release results show that the release of core PTH-Fc can be divided into rapid release in the early short time(0-5 d,up to 41.06% ± 0.06%)and subsequent slower release at a constant rate(6 After 30 days,it reached 70.09% ± 1.86%),and the release time of PTH-Fc in the membrane became longer;In vitro degradation experiments showed that with the prolongation of time,the surface of the film gradually became rough,and even some holes with uneven sizes appeared,and the fibers on the surface of the film were broken.(3)In vitro experiments showed that rat bone marrow mesenchymal stem cells(BMSCs)adhered to the membrane surface and grew well;Alizarin red staining showed obvious orange-calcified deposits;The results of the ALP activity assay showed that the ALP activity was significantly increased.Conclusion: The PTH-Fc/SF/PLCL three-dimensional nanofiber barrier film can be prepared by using the coaxial electrospinning technology.Its morphology is good,its mechanical properties are excellent,it has the ability of slow degradation and the slow release of the drug,and its biocompatibility is excellent.Has the ability to promote bone formation in vitro.This drug-guided three-dimensional nanofiber barrier film,which will be used to guide bone tissue regeneration technology combined with growth factors and drug release technology,is expected to become a new type of effective GBR barrier membrane for bone regeneration,and it is expected to be applied Bone tissue regeneration.