Comparison Of Osteogenesis Ability Between Canine Alveolarb-One-derived Stem Cell And Bone Marrow Mesenchymal Stromal Cells In Vivo

Objective To compare the osteogenic potential of iliac bone marrow mesenchymal stem cells(I-BMSCs)with alveolar bone marrow MSCs(Al-BMSCs)when combined with P-TCP in bone defect around implants as well as to investigate the osseointegration between tissue-engineered bone and dental implants in dogs.Methods The primary alveolar bone stem cells(Al-BMSC)and iliac stem cells(I-BMSC)were obtained and cultured,and the cell morphology was observed under the common inverted phase contrast microscope.Cells were cultured in DMEM containing 10%FBS,the third generation of Al-BMSCs and I-BMSCs were used to construct complexes with ?-TCP.Scanning electron microscopy(SEM)was used to observe the cells on the surface.Six Labrador dogs were extracted bilateral mandibular premolars,three implants were implanted.Bone defects(5mm × 5mm × 6mm,mesiodistal dimension × buccolingual dimension × depth)were constructed near the implant.The complexes were implanted.The experiments were divided into three groups:1.?-TCP ?Al-BMSC group,2.p-TCP ? I-BMSC group,3.simple p-TCP group.Antibiotics were injected once every 3 days after surgery.The fluorescent marker tetracycline(25 mg/kg)was intramuscularly injected at the 4th week,the fluorescent marker calcein(20 mg/kg)was injected at the 8th week,and fluorescent marker alizarin red(30 mg/kg)was injected at the 10th week after surgery.12 weeks after the surgery,the specimen was harvested and analyzed histologically.Statistical analysis was performed,P<0.05 was considered statistically significant.Results The common inverted phase contrast microscope showed that Al-BMSCs and I-BMSCs had good growth state,and the cell morphology was uniform,which was long spindle shape.Scanning electron microscopy(SEM)was used to observe the surface of the complexes,showed the surface of the material had cells adhesion,which were in good condition.The laser scanning confocal microscopy showed that the fluorescence expression of Al-BMSC+?-TCP group and I-BMSC+?-TCP group was significantly higher than that of ?-TCP at the different time points after bone defect repair(P<0.05).From the 4th to the 8th week after surgery,the expression of tetracycline in the Al-BMSC+P-TCP group was significantly higher than that in the other two groups(P<0.05).From the 8th week to 12th week after surgery,there was no statistically significant difference in the expression of fluorescent between Al-BMSC+?-TCP group and I-BMSC+?-TCP group(P>0.05).Histological analysis of VG staining showed that the area of new bone tissue in Al-BMSC+?-TCP group and I-BMSC+?-TCP group was significantly higher than that in ?-TCP group(P<0.05).The area of new bone formation in I-BMSC+?-TCP group was higher than that in Al-BMSC+P-TCP group,but there was no significant difference(P>0.05);while BIC with obvious statistical difference was higher in the ?-TCP+ I-BMSCs group than that in the p-TCP+ Al-BMSCs group(P<0.05).Conclusions Both Al-BMSCs and I-BMSCs have the ability of osteogenesis.More BIC could be obtained by using I-BMSCs.