| Objective:As the most common oral diseases in the world, periodontal disease is the main factor of inducing tooth loss among adults. It is considerably detrimental to oral and general soundness of human and it is of a serious incidence around the world. The main pathological change can be attributed to the absorption of periodontal tissue. How to achieve the regeneration of periodontal support loss is becoming a hot spot in periodontal tissue engineering field.Core binding factor al (Cbfal) is a very important transcription factor in bone development. It is not only the key factor in osteoblast differentiation and osteoblast function, but also plays an key role in tooth development which is the specific transcription factor of the odontogenic cell lines. Cbfal operates by binding to the osteoblast-specifics-acting element2(OSE2), which was founded in the regulatory region of all main osteoblast-related genes controlling their expression during bone formation. Special AT-rich sequence binding protein (Satb)2is a nuclear matrix protein, which can cooperate with significant transcription factors of osteogenic differentiation such as Cbfal. Osx, Atf4and so on. It shows the profound influence in the differentiation of osteoblasts as well as dental and craniofacial development. Bone formation is a complicated process, regulated by a variety of transcription factors and bone formation gene. So combining with two transcription factors in various ways which plays a different regulating role to bone formation and osteoblast differentiation is conceived to study the influence on signaling molecules in bone tissue engineering then thereby provides technical support for bone tissue engineering during the choice of signaling molecules.Specifically, lentivirus vector is capable of efficient and lasting expression of target genes, and our research team has successfully obtained the high titer lentivirus of co-expression and respective expression vectors such as Plenti6.3-Cbfal-IRES-Satb2(pL-cs)、Plenti6.3-Cbfal-IRES-Egfp (pL-c)、Plenti6.3-Satb2-IRES-Egfp (pL-s) and Plenti6.3-IRES-Egfp (pL-e) in the early stage. This experiment study aims to establish a Wistar rat mandible defect model, investigate the effect of Cbfal and Satb2gene when repairing the defect by lentivirus vector.Methods:Forty male Wistar rats of weighting160-180g were used in this experiment. They were divided into five groups:control group, pL-c group, pL-s group, pL-cs group and pL-e group randomly. The time interval was prescribed as two weeks and four weeks during which five groups with4rats in each are dispatched. The bilateral mandibular periodontal defect model was used in the experiment. This model size was a5mm X4mm X lmm which defect was directly to the root surface.25μl physiological saline, containing the same amount of virus pL-c, pL-s, pL-cs, pL e with a final volume of25μl lentivirus suspension gelation were composited with sponge. Then, they were transplanted into the animal mandibular defect respectivly.After2and4weeks, the specimens were obtained. Cbfal and Satb2immune histochemical staining and statistical analysis were used to detect the over-expression of gene by decalcified tissue sections.HE staining, Masson staining, bone sialoprotein (bone sialoprotein, BSP), osteopontin (osteopontin, OPN) immune histochemical staining and histological measurement were used to evaluate the bone defects and repair of periodontal defects. Meanwhile, Masson staining of hard tissue sections were used to observe the repair of bone defects and periodontal defects.Results:The results of Cbfal and Satb2immune histochemical staining and statistical analysis show that third-generation lentiviral system is capable of infecting cells of the defect. The genes are over expressed successfully. The expression of exogenous gene Cbfal and Satb2increase obviously, which is2to6times as much as infected empty vector group and control group (P<0.05).HE staining, Masson staining, BSP, OPN immunohistochemical staining and statistical analysis results show that periodontal repair and regeneration capacity of pL-c, pL-s and pL-cs groups is all stronger than the control group and the pL-e group (P<0.05). The osteogenic ability of pL-c and pL-s groups is stronger than pL cs group after2weeks, and pL-c group has the strongest osteogenic ability (P<0.05). Though the bone formation ability of pL-c is still stronger than the pL s group after4weeks, but both of them are weaker than the pL-cs group (P<0.05).Conclusions:Lentivirus expression systerm of Cbfal and Satb2genes both have strong periodontal repair and regeneration ability. The bone repair capacity of Cbfal is more obvious than Satb2after2and4weeks. The bone repair capacity of Cbfal-Satb2is more obvious than Cbfal and Satb2gene after4weeks. |
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