OBJECTIVETo investigate the green fluorescent protein (GFP) expression and the bionomics of skeletal muscles satellite cells (SMSCs) in vitro in GFP transgenic mouse.METHODSThe newborn transgenic mice were acquired to separate skeletal muscles satellite cells with enzyme digestion method. Cells were cultured and subcultured in vitro. Morphological observation, growth curve were investigated to evaluate the proliferation and differentiation characteristics of skeletal muscles satellite cells, fluorescence microscope was used to observe the GFP expression. The cells were identified by immunocytochemical stain. In the basis of identification of anti-sarcometric actin anti-body, the combination of anti-desmin antibody and DAPI (4, 6-diamidino-2-phenylindole) were used to detect the purification of skeletal muscles satellite cells.RESULTSImmunocytofluorescence suggested the good retain of GFP fluorescence in skeletal muscles satellite cells. The cells showed strong proliferative ability and they were positive with immunocytochemical stain of anti-sarcometric actin antibody and anti-desmin antibody. The combination of anti-desmin and DAPI stain can be used to determine the purification of SMSCs.CONCLUSIONSkeletal muscles satellite cells cultured in vitro showed strong proliferation and differentiation ability. They are fit to construct the cell bank of tissure engineering and to be a useful tool to explore cells fate after transplantation since these cells retain the expression of GFP.

OBJECTIVETo observe the ability of induced ectopic bone using skeletal muscles satellite cells (SMSCs) from newborn green fluorescence protein (GFP) transgenic mice mediated by Ad-BMP2.METHODSTransplantation of SMSCs transduced with Ad-BMP2 into back lamb muscles of subfascia in wildtype 129sv mice with a complex of collagen scaffords, then the tissue histologic examination, X ray plain film, fluorescence microscopy were used.RESULTSTransplantation of SMSCs transfected with Ad-BMP2 into back lamb muscles of subfascia generated ectopic bone formation involving GFP-positive osteoblasts and osteocytes 2 weeks and mature bone formation 4 weeks after transplantation. SMSCs non-transfected with Ad-BMP2 failed to induce ectopic bone formation.CONCLUSIONSMSCs retain differentiation potentitality into osteoblasts in response to Ad-BMP2. They are useful tools for analyzing the process of osteoblast differentiation in vivo after transplantation.

Objective: To assess the ability of inducing skeletal muscle satellite cells(SMSCs) into osteoblast-like cells,and to approach the possibility of treating skeletal muscle satellite cells as a new seed cell for bone tissue engineering.Method: The skeletal muscles of newborn Wistar rats were acquired to separate skeletal muscles satellite cells with enzyme digestion.Cells are cultured and subcultured in vitro.After being transfected by Ad-BMP2 gene,cells were examined by osteoblastic marker's activity test,cytochemical staining and mineralized capability in vitro.Results: After the cells were transfected,there was a significant increase in ALP activity(P0.01),the cells were faint positive with immunocytochemical stain of osteocalcin and type I collagen.Calcium nodules were seen after cells were cultured for 21 days.Conclusion: SMSCs have the capacibility to be differentiated into osteoblast and to be regarded as the seed cells for bone tissure engineering.

2001年8月～2005年8月,我们采取不用手术方式对58例颌骨缺损患者进行了修复重建,效果良好.现报告如下.临床资料:本组58例患者中,男38例,女20例;年龄11～66岁.恶性肿瘤42例(牙龈癌21例,颌骨肉瘤10例,中央性颌骨癌11例),良性肿瘤16例(成釉细胞瘤13例,颌骨黏液瘤2例,角化囊肿1例).肿瘤切除术后上颌骨缺损6例,下颌骨缺损52例.

Objective: To investigate the culture method in vitro and the bionomics of masticatory muscle satellite cells.Methods: The masticatory muscle satellite cells were obtained from newborn green fluorescence protein(GFP) transgenic mice by enzyme digestion,then they were cultured and sub-cultured in vitro.The proliferation and differentiation characteristics of masticatory muscles satellite cells were observed by inverted microscope and fluorescence one,and they were detected by growth curves,and the cells were identified by immunocytochemical staining using anti-sarcometric actin monoclonal antibody.Results: The masticatory muscle satellite cells were successfully separated in vitro by enzyme digestion and strongly proliferated,and they were faint positive with immunocytochemical stain of α-sarcometric actin.Conclusion: Masticatory muscle satellite cells cultured in vitro strongly proliferate and differentiate,and can be used to construct the cell bank of tissue engineering.

OBJECTIVE:To investigate the possibility of adipose-derived stromal cells (ADSCs) transfeced by adenovirus containing human bone morphogenetic protein-2 (Ad-hBMP-2) gene and their osteogenic potential.METHODS:ADSCs were obtained from inguinal fat tissue of 4 weeks old SD rats. After exposure to adenovirus containing green fluorescent protein(Ad-GFP), fluorescent microscope was used to observe gene transfection effect once 12 hours. After transfected with Ad-hBMP-2, cytochemistry, immmucytochemistry and Western blot were used to examine the expression of alkaline phosphatase (ALP), osteocalcin (OC) and hBMP-2.RESULTS:After exposed to Ad-GFP 12 hours, 52% ADSCs were observed being transfected and 48 hours later reached 95%. The double number time belonged after transfecting with Ad-hBMP-2, and cytochemistry, immucytochemistry and Western blot examines indicated positive results of ALP, OC, hBMP-2 after 48 hours.CONCLUSION:Adipose tissue contains abundant ADSCs which could be transfected as gene vectors by adenovirus, ADSCs transfected with Ad-hBMP-2 can convert to ostoeblasts, and can act as a kind of seed cells for osteo-tissue engineering.

Objective:To investigate the effect of different inner crown materials on periodontal tissue by means of measuring the amount level of AST and ALP in gingival crevicular fluid(GCF)after PFM restoration.Methods:The anterior teeth of 26 patients by random were restored by Ni-Cr alloy PFM,Ti alloy PFM and Au-Pt alloy PFM.GCF was collected from labial and lingual of mesial site and distal site before the tooth preparation by filter paper.At the point of 1 month and 3 months after cementation,GCF was collected at the same sites.The level of AST and ALP were detected by automated chemical analyzer,and analyzed the variation between the pre-restoration and post-restoration.Results:Compared to pre-restoration,all indexes have no conspicuous difference among three groups after 1 month;Ni-Cr alloy group have no difference with Ti alloy group after 3 months.However,Au-Pt alloy group has conspicuous difference with Ni-Cr alloy group and Ti alloy group respectively.Conclusion:Non-noble metal has badnon-favorable effect on the periodontal tissue.It canis easily and correctly to observe the effect of periodontal tissue dynamicallyin the dynamic state after PFM restoration by means of measuring the amount of AST and ALP in GCF.

富血小板血浆(PRP)是全血经过离心分离而得到的血制品,含有全血中70%以上的血小板,现已知PRP是自体生长因子库,富含多种生长因子,如血小板源性生长因子(PDGF)、转化生长因子(TGF)、成纤维细胞生长因子(FGF)和胰岛素生长因子(IGF)等[1,2].

观察种植界面微区元素在种植界面形成中的作用.用电子探针X射线能谱分析方法对界面进行百分含量测定.结果表明,钛合金种植体植入后,12周内,钙元素与磷元素逐渐增加并达到峰值,以后趋于平稳,钛元素含量植入后2周较高,以后逐渐减少.种植体与机体组织之间只有极薄的一层纤维膜,已达到骨结合标准.钙元素与磷元素在骨结合界面形成过程中起主导作用,钛合金种植体植入后,短期内会释放游离离子,但不足以影响骨结合界面的形成.