The Research About Platelet-rich Fibrin To Construct Bone Tissue Engineering

Objective:(1)Explore the structure characteristics of platelet-rich fibrin from the beagles.(2)Evaluate the possibility of platelet-rich fibrin to be bone tissue engineering scaffold.(3)Evaluate the effect of platelet-rich fibrin combine BMSCs to construct bone tissueengineering in repairing bone defect.(4)Screening bone tissue engineering seed cells which more suitable in early osteogenicafter platelet-rich fibrin acting on in vitro.Methods:(1) Rapid extraction20ml blood from the beagle neck vein then centrifugal at3000rpmto get platelet-rich fibrin, after using the scanning electron microscope and HE staining toobserve its structure.(2) Extracting5ml bone marrow from Anterior superior iliac spine to culture and expand.Combined platelet-rich fibrin and measured the proliferation of bone marrowmesenchymal stem cells, and early osteogenesis ability.(3) Nine Beagles unplug bilateral lateral incisor to build buccal bone wall missing toothextraction sites were randomized groups: the experimental group A: blood clots group (n=6); experimental group B: separate PRF group (n=6); experimental group C:tissue-engineered bone group (PRF combined bone marrow mesenchymal stem cells)(n=6). All Beagles no abnormality after surgery. After surgery, six weeks and twelve weeksobserved by cone-beam CT to evaluate osteogenesis height and bone density. (4) Using a modified tissue culture techniques to isolate and expand beagles deciduousdental pulp cells and permanent dental pulp cells and to identify the multi-directionalthrough differentiation. The platelet-rich fibrin (PRF) acting on deciduous dental pulpcells, permanent dental pulp cells, at1,4,7,11days measured cell proliferation, alkalinephosphatase semi-quantitative, real-time quantitative PCR-related osteogenic genes,21days evaluation of the number of calcium nodules.Results:(1)Platelet-rich fibrin can be easily obtain from beagles, In the performance of itsmicroscopic structure of the porous mesh stent and contains a large number of whiteblood cells.(2)Platelet-rich fibrin in vitro with bone marrow mesenchymal stem cells can promote theproliferation and early osteogenesis.(3)Beagles buccal bone wall deficient bone repair experiments, after six weeks and twelveweeks, cone-beam CT evaluate the bone height and bone mineral density, PRFcombined bone marrow mesenchymal tissue engineered bone showed most significant.There are significant differences in six weeks, missing buccal bone wall is almostcompletely repaired. Bone height (2.63±0.57) and bone mineral density (765.19±59.70)compared blood clots group (5.65±2.91,599.08±53.88) and PRF group (4.14±1.16,644.76±65.39) was significantly higher (p <0.05).(4) Deciduous dental pulp cells and permanent dental pulp cells growth vigorous,multi-differentiation identified two cells own the ability of into bone, cartilage andadipocyte. PRF in vitro effects on deciduous dental pulp stem cells, permanent dentalpulp stem cells and bone marrow mesenchymal stem cells, can promote the proliferationcalcification and osteogenic differentiation of three cells, compare the control group hadsignificant difference (P <0.05). Real-time quantitative PCR-related osteogenic geneexpression in the role of PRF express in earlier periods, and the difference was significant (P <0.05). In the dental pulp cells, deciduous dental pulp stem cells comparedto permanent dental pulp stem cells are more sensitive to the role of the PRF, owing thestronger ability to self-mineralized.Conclusions:(1) Platelet-rich fibrin is a fiber mesh scaffolds without toxic to cells and its contains lotsof leukocyte.(2) Platelet-rich fibrin can promote cell proliferation and osteogenic differentiation whichbe very useful as a scaffold for bone tissue engineering.(3) PRF can be used as scaffold materials combined BMSCs in early repairing of buccalalveolar wall defects.(4) Deciduous dental pulp cells can be used as a better seed cells to compositeplatelet-rich fibrin.