Research Of The Application Of CGMs Scaffold Filled With Osteoblasts In Skull Repair

Bone is one of the important tissues that constitute human body structure.Bone tissue disorders and injuries caused by various reasons in life can seriously affect the quality of life of individuals.In the natural state,when the bone tissue defect is more serious,the natural bone is difficult to complete the self-repair.In clinical practice,many postoperative neurosurgery patients are faced with a large area of skull defects,which will cause great adverse effects on the patients’ physiology and psychology.At present,the main clinical methods for the treatment of skull defects are autologous skull flap repair and artificial titanium mesh repair.Autologous bone repair is limited by whether the autologous bone flap is well preserved,and the bone flap will be absorbed by the body to some extent after the repair,which has a great negative impact on the surgical effect.However,artificial titanium mesh repair has some disadvantages such as high cost and relatively large inflammatory response.Fortunately,bone tissue engineering has developed rapidly in recent years.One of the major goals of bone tissue engineering is to create scaffolds that mimic the natural extracellular matrix(ECM)and provide a three-dimensional structure for cell adhesion and proliferation.At present,the materials required for scaffold synthesis are mainly some natural derivative materials and some synthetic materials.The use of natural materials in the preparation of scaffolds can significantly reduce the inflammatory reaction and facilitate the natural degradation of the body.Among many natural materials,gelatin,chitosan and other natural materials are widely used.Object:Gelatin,as a natural biological material with the highest similarity to collagen,is widely used in tissue engineering.However,gelatins have poor spatial stability,fast dissolution in vivo,and poor cell affinity,which results in the inability of cell proliferation and proliferation in gelatins.Chitosan is a kind of natural polysaccharide.Because of its strong porous structure,natural antibacterial properties and the promotion of osteoblast matrix mineralization,it has become an ideal material for bone tissue engineering.Therefore,the Chitosan gelatin microspheres(CGMs)are being prepared by mixing Chitosan and Gelatin,and their biocompatibility has been tested by various means.Methods : Bone marrow mesenchymal stem cells(BMSCs)were extracted and differentiated into osteoblasts in vitro.Fabricating the Chitosan gelatin microspheres(CGMs).Cell and CGMs structure were observed under inverted phase contrast microscope.MTT assay,hemolysis test,dead/living cell staining and DAPI staining were used to detect the biocompatibility between CGMs scaffolds and osteoblasts.PCR was performed to evaluate the osteogenic effect of osteoblasts in scaffolds.Results : Osteoblasts were observed to grow well in the extract under microscope.Inverted phase contrast microscope showed that the prepared CGMs stents were about200 um in diameter and orange in color.The hemolysis test results showed that the hemolysis rate of CGMs scaffold extract was 2.08%(< 5%).MTT assay showed that there was no significant difference in absorbance between the control group and the extract group(P >0.05),and the cytotoxicity level was level 1.DAPI staining and dead/living cell staining showed that osteoblasts proliferated well on the scaffold.PCR results showed that osteoblasts survived well on the CGMs scaffold,and the osteogenesis was normal.Conclusion:CGMs scaffold has good biocompatibility and is expected to be a new type of bone tissue engineering material.