The Effect Of 3D Printed β-tricalcium Phosphate Combined With Low Frequency Pulsed Electromagnetic Field On The Repair Of Rat Skull Defect

Trauma,infection,cancer,and congenital diseases can lead to bone defects.The treatment of bone defects often requires autologous bone transplantation or allogeneic bone transplantation,and these treatment methods have certain defects.Bone tissue engineering that contains 3D printing technology and biological materials can make up for these defects to a certain extent,which is of great significance to the treatment of bone defects.In addition,pulse electromagnetic fields(PEMFs)can promote bone regeneration.However,there is currently a lack of research on PEMF for the treatment of bone defects.The combination of the two may provide a new approach for the treatment of bone defects.Purpose:1.The effects of 3D-printed scaffold and PEMF on proliferation and osteogenic differentiation of rat adipose-derived stem cells(rADSCs)were studied2.To investigate the effect of combination of β-TCP scaffold and PEMF irradiation on critical bone defect repair in rats.Method:1.Using 3D printing technology to customize theβ-TCP scaffold according to the design scheme.2.3D-printedβ-TCP scaffolds were used to observe the surface morphology by scanning electron microscopy.3.Live/dead cell staining experiments were performed to evaluate the biocompatibility and cytotoxicity of theβ-TCP scaffold after the cultivation of rADSCs onβ-TCP scaffolds for 24,48,72 h.4.The proliferation of rADSCs was accessed by using the Cell Counting Kit-8(CCK-8)method after cultured for 1,4,7 days under four different culture conditions,respectively.5.After 14 days of incubation of rADSCs under four different culture conditions,the activity of alkaline phosphatase(ALP)and ALP staining were used to evaluate the osteogenic differentiation process of rADSCs.6.The expression of osteogenesis-related genes in rADSCs was accessed by RT-qPCR after 7,14,21 days of culture under four different conditions.7.Micro-computed tomography(mCT),BMD analysis and hematoxylin-eosin(H-E)staining were used to evaluate the repair of rat cranial bone after modeling the skull defect model of SD rats with implanted β-TCP scaffolds and feeding 4,8,12 w in two ways.Results:1.The surface morphology,size and pore diameter of the β-TCP scaffold were observed by scanning electron microscope,which also showed its high porosity2.The results of the Live/Dead cell staining experiment suggest that the ratio of live cells to dead cells of rADSCs on theβ-TCP scaffold is high andβ-TCP has good biocompatibility.3.CCK-8 showed thatβ-TCP scaffolds could significantly increase the proliferation of rADSCs,while PEMF significantly increased the proliferation rate of rADSCs in the middle and late stages of proliferation.4.ALP activity detection and ALP staining suggested that bothβ-TCP and PEMF could promote osteogenic differentiation of rADSCs5.Osteogenic-related gene expression detection suggested that β-TCP and PEMF could improve the expression of ALP,Runx2 and OPN genes during osteogenic differentiation of rADSCs6.The results of 3D reconstruction of Micro CT and BMD analysis suggest that bothβ-TCP and PEMF can promote the repair of skull defects in SD rats.The combination of the two can achieve the best results.7.The results of HE staining suggested that the defect of transplantedβ-TCP scaffold was well grown,and PEMF also promoted the repair of skull defect.Conclusions1.The high-porosityβ-TCP scaffolds manufactured by the 3D printing process have good cytocompatibility.2.β-TCP scaffold and PEMF treatment could promote the proliferation of rADSCs.3.β-TCP scaffold and PEMF treatment could promote the osteogenic differentiation process of rADSCs.4.β-TCP scaffolds and PEMF treatment can promote the repair of skull defects in SD rats.