自体颏部骨移植并种植体修复前牙槽骨缺损的应用研究

目的:评价自体颏部骨移植方法在前牙槽骨缺损植骨及种植的效果。方法:对16例前牙骨缺损的患者进行自体颏部骨移植,分别于植骨12～16周后植入种植体,共植入18枚种植体,术后16～20周完成修复治疗。结果:16例自体骨移植患者中,1例出现术后感染,经局部冲洗换药后后愈合良好,其余患者移植骨均已成活,修复后种植体至今均无脱落,局部外形良好。结论:自体颏部骨移植能有效恢复局部牙槽外形,为前牙美学修复提供良好的基础。     

无机诱导成骨材料的研究进展

临床上骨缺损十分常见,常需要进行骨移植来修复。在牙种植术中,由于多种原因造成牙槽嵴萎缩,骨量不足,限制了牙种植术的应用。对于骨量不足或拔牙后即刻种植的情况可以采取骨增量技术增加牙槽嵴的高度和丰满度,提高种植成功率。本文主要阐述近年来无机诱导成骨材料的研究进展,并对其将来的研究方向予以展望。     

拔牙位点保存技术的现状与发展

随着当代医学的发展,口腔种植技术被越来越多的患者所接受。但是牙齿拔除后牙槽骨会发生吸收,周围缺乏固定和支持种植体的骨组织,会对种植体的稳定性产生重大的影响,同时牙槽骨的不足对种植后期的美容修复也会产生不利的影响。牙槽骨的不足是我们当前所面临的最大问题。拔牙位点保存技术,是减少拔牙后牙槽骨吸收,促进牙槽窝内新骨形成的一系列临床治疗方法。常用的拔牙位点保存技术有即刻种植、植骨等。本综述就拔牙位点保存技术的概念、常用的方法、现状与未来的发展等做一综述,为临床上拔牙位点保存技术的应用提供理论依据。     

OSFE植骨与不植骨种植修复的临床效果对比研究

目的:比较上颌窦挤压内提升(OSFE)植骨与不植骨种植修复的临床效果。方法:选择上颌后牙区种植修复的35例患者,其剩余牙槽嵴高度(RBH)为4~8 mm,共植入43颗种植体。A组16例患者为植骨组,20个种植位点,牙槽骨可用骨高度平均(5.87±1.19)mm,植入人工骨粉后植入种植体;B组19例患者为不植骨组,23个种植位点,缺牙区牙槽骨可用骨高度平均(5.67±1.10)mm,上颌窦提升后直接植入种植体。6个月后行二期手术完成修复。采用临床检查、X线检查及视觉模拟评分法(visual analogue scale,VAS)进行效果评价。结果:两组病例的牙槽嵴高度差异比较无统计学意义。在平均约36.7个月的随访期内,A组种植体的存留率为100%(20/20),B组中有1枚种植体因咬合力过大及口腔卫生较差脱落,种植体的存留率为95.6%(22/23),两组病例的存留率比较无统计学差异。两组患者的VAS值比较亦相当。所有种植体骨结合良好,种植体周围软组织无炎症,种植义齿咀嚼功能良好。结论:在严格控制适应症、准确掌握种植技巧的前提下,RBH在4~8 mm之间的病例无需额外植入骨代替材料即可取得理想的修复效果,简化了手术的操作,减少了手术的风险和创伤,节省了手术的时间和费用,易被患者接受。     

前牙美学区实施即刻种植及延期种植对患者唇侧骨量变化的对比研究

目的：研究对比前牙美学区实施即刻种植及延期种植对患者唇侧骨量变化。方法：选择2012 年4 月至2013 年4 月在我院 接受上颌前牙区的单颗种植手术治疗的患者96 例作为研究对象。根据数字法随机分成即刻种植组48 例（48 颗ITI种植体）与延 期种植组48 例（48 颗ITI种植体）。对比两组术后唇侧骨量情况,唇侧骨的改建情况以及术后1 年的PES评分。结果：即刻种植组 术后1 年的唇侧骨高度较术后即刻显著更高,差异有统计学意义（P＜0.05）。即刻种植组与延期种植组术后1 年水平向骨改建的 种植体冠方C 及中段M 较术后即刻显著更低,差异均有统计学意义（均P＜0.05）。即刻种植组在术后1 年的唇侧牙槽嵴的高度 降低水平明显大于延期种植组,且对于水平向骨改建方面,两组种植体中段M的差异具有统计学意义（P＜0.05）。两组在术后1 年的PES 评分上对比,差异无统计学意义（P＞ 0.05）。结论：即刻种植并未能阻止患者牙槽嵴生理吸收,但种植的美学效果亦较 好。临床运用时应加以关注,掌握好适应证,从而获得最佳疗效。值得重视。     

PRP 与CPC 应用于拔牙后种植位点保存的实验研究

目的:探讨磷酸钙骨水泥(CPC)与富血小板血浆(PRP)混合物应用于拔牙后种植位点保存的可行性。方法:拔除6只犬下颌双侧第三切牙与一侧第一切牙,并于每只犬的3个拔牙窝颊舌侧骨壁分别制造2 mm×2 mm×3 mm缺损,随机在两个牙槽窝内分别植入CPC或CPC与PRP的混合物,第三个不加处置作对照,在术后4、8、12周各处死两只动物,行大体、X线、显微镜观察,比较各组的新骨生成情况。结果:术后4周时CPC/PRP混合物组骨形成早于其他组,8、12周时CPC/PRP混合物组新骨生成明显优于CPC组和对照组。结论:CPC混合PRP可促牙槽窝新骨生成并能保证成骨的高度和颊舌向宽度,从而为后期种植义齿修复提供了必要的条件。     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

人工骨表面覆盖口腔修复膜对颌骨囊性缺损人工骨植入术患者植入腔感染的预防作用

目的:探讨人工骨表面覆盖口腔修复膜对颌骨囊性缺损人工骨植入术患者植入腔感染的预防作用。方法:将我院2012年4月～2016年2月收治的78例颌骨囊性缺损患者按治疗时间分为对照组38例与观察组40例,均行开窗人工骨植入术,其中对照组采取常规抗感染,观察组采取人工骨表面覆盖口腔修复膜。比较两组的囊腔体积、面积缩减率、人工骨植入量、骨厚度及植入腔感染率。结果:两组的囊腔体积、面积缩减率及人工骨植入量比较差异无统计学意义(P0.05)。但观察组的骨厚度明显大于对照组(P0.05),植入腔感染率明显低于对照组(P0.05)。结论:在开窗人工骨植入术的基础上使用口腔修复膜不仅能引导骨组织再生,而且能够显著降低植入腔的感染率。     

Shh信号参与调控BMP9诱导的间充质干细胞成骨分化

目的：观察sonic hedgehog（Shh）信号通路在骨形态发生蛋白9（BMP9）诱导的小鼠间充质干细胞（MSCs）C3H10T1/2和C2C12成骨分化中的作用,并初步探讨其作用机制。方法：Shh信号通路抑制剂Cyclopamine和激活剂Purmorphamine以及过表达Shh腺病毒分别作用于BMP9处理的C3H10T1/2和C2C12细胞,碱性磷酸酶（ALP）检测早期成骨指标ALP,茜素红S染色检测晚期成骨指标钙盐沉积,RT-PCR检测Shh信号相关基因以及成骨关键转录因子的表达,Western blot检测Shh的表达,荧光素酶报告基因检测Smad1/5/8的转录调控活性。结果：BMP9促进Shh信号相关基因的表达,激活Shh信号可增强BMP9诱导的C3H10T1/2和C2C12细胞早晚期成骨分化并促进了BMP9诱导的Smad荧光素酶活性,抑制Shh信号后作用相反。结论：激活Shh信号通路可促进BMP9诱导的小鼠MSCs成骨分化,抑制其活性后作用相反。     

BMP2 Genetically Engineered MSCs and EPCs Promote Vascularized Bone Regeneration in Rat Critical-Sized Calvarial Bone Defects

Current clinical therapies for critical-sized bone defects (CSBDs) remain far from ideal. Previous studies have demonstrated that engineering bone tissue using mesenchymal stem cells (MSCs) is feasible. However, this approach is not effective for CSBDs due to inadequate vascularization. In our previous study, we have developed an injectable and porous nano calcium sulfate/alginate (nCS/A) scaffold and demonstrated that nCS/A composition is biocompatible and has proper biodegradability for bone regeneration. Here, we hypothesized that the combination of an injectable and porous nCS/A with bone morphogenetic protein 2 (BMP2) gene-modified MSCs and endothelial progenitor cells (EPCs) could significantly enhance vascularized bone regeneration. Our results demonstrated that delivery of MSCs and EPCs with the injectable nCS/A scaffold did not affect cell viability. Moreover, co-culture of BMP2 gene-modified MSCs and EPCs dramatically increased osteoblast differentiation of MSCs and endothelial differentiation of EPCs in vitro. We further tested the multifunctional bone reconstruction system consisting of an injectable and porous nCS/A scaffold (mimicking the nano-calcium matrix of bone) and BMP2 genetically-engineered MSCs and EPCs in a rat critical-sized (8 mm) caviarial bone defect model. Our in vivo results showed that, compared to the groups of nCS/A, nCS/A+MSCs, nCS/A+MSCs+EPCs and nCS/A+BMP2 gene-modified MSCs, the combination of BMP2 gene -modified MSCs and EPCs in nCS/A dramatically increased the new bone and vascular formation. These results demonstrated that EPCs increase new vascular growth, and that BMP2 gene modification for MSCs and EPCs dramatically promotes bone regeneration. This system could ultimately enable clinicians to better reconstruct the craniofacial bone and avoid donor site morbidity for CSBDs.     

胰岛素对糖尿病兔牙槽骨缺损修复的影响

目的：探讨胰岛素对糖尿病兔牙槽骨缺损修复治疗的效果,为糖尿病所致的牙周炎提供临床治疗的依据,方法：40只大耳白兔随机分为4组：A组制备健康兔的牙槽骨缺损;B组为胰岛素组,制备健康兔牙槽骨缺损后,用胰岛素治疗;C组为糖尿病组,制备糖尿病兔牙槽骨缺损;D组为糖尿病胰岛素治疗组,制备糖尿病兔牙槽骨缺损后,用胰岛素治疗。每组10只,缺损制备后4、8周各处死5只,对各组成骨情况进行组织学观察及测定。结果：组织学观察A、B、D组修复区可见大量新骨形成,以B组为显著;C组仅见少许成骨,多为纤维组织。新生骨面积比和成骨细胞数在4、8周时均为D组大于c组,B组大于A组,组间差异有统计学意义（P〈0．05）。证明应用胰岛素促进糖尿病兔缺损牙槽骨形成新骨的效果明显。结论：胰岛素能够促进糖尿病兔牙槽骨缺损的戍骨,为,临床上治疗糖尿病并发牙周炎提供一种新的手段。     

Bone morphogenetic protein 2 promotes osteogenesis of bone marrow stromal cells in type 2 diabetic rats via the Wnt signaling pathway

Type 2 diabetes mellitus impairs osteogenesis in bone marrow stromal cells (BMSCs). Bone morphogenetic protein 2 (BMP2) has been extensively applied for bone defect restoration and has been shown to activate the Wnt signaling pathway. The objective of this study was to investigate the effects of BMP2 on the cell proliferation and osteogenesis of type 2 diabetic BMSCs in rats and explore whether BMP2 induced osteogenesis via the stimulation of Wnt signaling pathway. The cell experiments were divided into DM (diabetic BMSCs), BMP25 (induced with 25 ng/ml BMP2), BMP100 (induced with 100 ng/ml BMP2) and BMP25  + XAV groups. All cells with or without the different concentrations of BMP2 were cultured under the same experimental conditions. The in vitro results indicated that BMP2 enhanced cell proliferation by 130%–157% and osteogenic differentiation by approximately two-fold in type 2 diabetic BMSCs. The expression levels of β-catenin, cyclin D1, Runx2 and c-myc related to the Wnt signaling pathway were also upregulated from 180% to 212% in BMP2-induced type 2 diabetic rat BMSCs, while the level of GSK3β decreased to 43%. In BMP2-induced type 2 diabetic BMSCs with calcium phosphate cement (CPC) scaffolds for osteoblast study in vivo, the appearance of newly formed bone dramatically increased to 175% compared with type 2 diabetic BMSCs. These data demonstrated that BMP2 enhanced bone regeneration in diabetic BMSCs by stimulating the Wnt signaling pathway with the accumulation of β-catenin and the depressed expression of GSK3β. Diabetic BMSCs associated with BMP2 might be a potential tissue-engineered construct for bone defects in type 2 diabetes mellitus.     

The Angiopoietin-1 Variant COMP-Ang1 Enhances BMP2-Induced Bone Regeneration with Recruiting Pericytes in Critical Sized Calvarial Defects

Craniofacial bone defects are observed in a variety of clinical situations, and their reconstructions require coordinated coupling between angiogenesis and osteogenesis. In this study, we explored the effects of cartilage oligomeric matrix protein-angiopoietin 1 (COMP-Ang1), a synthetic and soluble variant of angiopoietin 1, on bone morphogenetic protein 2 (BMP2)-induced cranial bone regeneration, and recruitment and osteogenic differentiation of perivascular pericytes. A critical-size calvarial defect was created in the C57BL/6 mouse and COMP-Ang1 and/or BMP2 proteins were delivered into the defects with absorbable collagen sponges. After 3 weeks, bone regeneration was evaluated using micro-computed tomography and histologic examination. Pericyte recruitment into the defects was examined using immunofluorescence staining with anti-NG2 and anti-CD31 antibodies. In vitro recruitment and osteoblastic differentiation of pericyte cells were assessed with Boyden chamber assay, staining of calcified nodules, RT-PCR and Western blot analyses. Combined administration of COMP-Ang1 and BMP2 synergistically enhanced bone repair along with the increased population of CD31 (an endothelial cell marker) and NG2 (a specific marker of pericyte) positive cells. In vitro cultures of pericytes consistently showed that pericyte infiltration into the membrane pore of Boyden chamber was more enhanced by the combination treatment. In addition, the combination further increased the osteoblast-specific gene expression, including bone sialoprotein (BSP), osteocalcin (OCN) and osterix (OSX), phosphorylation of Smad/1/5/8, and mineralized nodule formation. COMP-Ang1 can enhance BMP2-induced cranial bone regeneration with increased pericyte recruitment. Combined delivery of the proteins might be a therapeutic strategy to repair cranial bone damage.     

Antagonistic and synergistic effects of bone morphogenetic protein 2/7 and all‐trans retinoic acid on the osteogenic differentiation of rat bone marrow stromal cells

The osteogenesis of bone marrow stromal cells (BMSCs) is of paramount importance for the repair of large‐size bone defects, which may be compromised by the dietary‐accumulated all‐trans retinoic acid (ATRA). We have shown that heterodimeric bone morphogenetic protein 2/7 (BMP2/7) could induce bone regeneration in a significantly higher dose‐efficiency in comparison with homodimeric BMPs. In this study, we evaluated the effects of ATRA and BMP2/7 on the proliferation, differentiation, mineralization and osteogenic genes. ATRA and BMP2/7 exhibited both antagonistic and synergistic effects on the osteogenesis of BMSCs. ATRA significantly inhibited proliferation and expression of osteocalcin but enhanced the activity of alkaline phosphatase of BMSCs. On day 21, 50 ng/mL BMP2/7 could antagonize the inhibitive effects of ATRA and significantly enhance osteogenesis of BMSCs. These findings suggested a promising application potential of heterodimeric BMP2/7 in clinic to promote bone regeneration for the cases with dietary accumulated ATRA.     

Adenovirus-mediated BMP2 expression in human bone marrow stromal cells

Recombinant adenoviral vectors have been shown to be potential new tools for a variety of musculoskeletal defects. Much emphasis in the field of orthopedic research has been placed on developing systems for the production of bone. This study aims to determine the necessary conditions for sustained production of high levels of active bone morphogenetic protein 2 (BMP2) using a recombinant adenovirus type 5 (Ad5BMP2) capable of eliciting BMP2 synthesis upon infection and to evaluate the consequences for osteoprogenitor cells. The results indicate that high levels (144 ng/ml) of BMP2 can be produced in non-osteoprogenitor cells (A549 cell line) by this method and the resultant protein appears to be three times more biologically active than the recombinant protein. Surprisingly, similar levels of BMP2 expression could not be achieved after transduction with Ad5BMP2 of either human bone marrow stromal cells or the mouse bone marrow stromal cell line W20-17. However, human bone marrow stromal cells cultured with 1 microM dexamethasone for four days, or further stimulated to become osteoblast-like cells with 50 microg/ml ascorbic acid, produced high levels of BMP2 upon Ad5BMP2 infection as compared to the undifferentiated cells. The increased production of BMP2 in adenovirus transduced cells following exposure to 1 microM dexamethasone was reduced if the cells were not given 50 microg/ml ascorbic acid. When bone marrow stromal cells were allowed to become confluent in culture prior to differentiation, BMP2 production in response to Ad5BMP2 infection was lost entirely. Furthermore, the increase in BMP2 synthesis seen during differentiation was greatly decreased when Ad5BMP2 was administered prior to dexamethasone treatment. In short, the efficiency of adenovirus mediated expression of BMP2 in bone marrow stromal cells appears to be dependent on the differentiation state of these cells.