The effect of demineralized intramembranous bone matrix and basic fibroblast growth factor on the healing of allogeneic intramembranous bone grafts in the rabbit

The aim here was to explore a new graft material that excludes the need to harvest autogenous bone from patients. Forty-two critical-size (10×15 mm) defects were created in rabbit mandibles bilaterally. Five groups of six defects each were grafted with autogenous endochondral (EC) bone, autogenous intramembranous (IM) bone, fresh-frozen allogeneic IM bone only, fresh-frozen allogeneic IM bone and demineralized bone matrix powder prepared from intramembranous bone (DBM_IM) only, and fresh-frozen allogeneic IM bone and basic fibroblast growth factor (bFGF) mixed with DBM_IM powder. The remaining defects were used as controls. Three weeks after surgery, the defects were retrieved for histological analysis. The amount of new bone formation was quantified by image analysis. No bone formed across the defect in the controls; 224% more new bone formed in defects grafted with composite allogeneic IM bone/DBM_IM than in those grafted with allogeneic IM bone alone (p<0.001); 550% more new bone was formed in defects grafted with composite allogeneic IM bone/DBM_IM/bFGF than in those grafted with allogeneic IM bone alone (p<0.001). The amount of new bone in the group receiving composite allogeneic IM bone/bFGF/DBM_IM was more than that in autogenous EC bone group, and very close to that in autogenous IM group. The results show that a composite of fresh-frozen allogeneic IM bone and bFGF in DBM_IM powder is a good graft material that warrants further clinical investigation.     

Zoledronic acid decreases gene expression of vascular endothelial growth factor and basic fibroblast growth factor by human epithelial cells

Delayed wound healing in patients taking bisphosphonates could result from decreased expression of growth factors, which are directly related to cell proliferation and migration. In this study, we evaluated the gene expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) by epithelial cells exposed to zoledronic acid 5 μmol for 48 h using real-time polymerase chain reaction. The gene expression of VEGF and bFGF by epithelial cells exposed to zoledronic acid decreased by 34% and 51%, respectively (p = 0.0001 and p = 0.0001). We conclude that zoledronic acid can decrease the expression of growth factors by epithelial cells.     

Giant cell granuloma of the jawbones – a proliferative vascular lesion? Immunohistochemical study with vascular endothelial growth factor and basic fibroblast growth factor

AIM: To estimate the angiogenic activity in central giant cell granuloma (CGCG) by immunohistochemical stains for vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). VEGF and bFGF immunoreactivity of the lesional mononuclear (MC) and giant (GC) cells was also investigated. METHOD: The study consisted of 41 cases of CGCG. Vascularity was quantified by microvascular volume (MVV) as determined by point counting. In five cases of CGCG, regions at the surrounding border, which demonstrated reactive vascular-rich inflammatory areas, served as control. Immunoreactivity of the MC and GC was assessed as the percentage of VEGF- and bFGF-positive cells from the total number of the respective cell type. RESULTS: Within CGCG lesions the extent of angiogenesis was low; MVV did not exceed 5% for either VEGF (88% of lesions) or bFGF (78% of lesions). The mean MVV of VEGF- and bFGF-positive blood vessels was 2.9% +/- 2.4% and 3.46% +/- 2.35%, respectively, significantly lower than in the control areas (27.5% +/- 7.3% and 28.08% +/- 5.5%, respectively) (P = 0.043). VEGF-positive and bFGF-positive MC and GC were found in nearly all lesions and in less than half of the lesions, respectively. CONCLUSION: The low mean MVV of VEGF- and bFGF-positive blood vessels implies low angiogenic activity, which does not support the designation of CGCG as a true proliferative vascular lesion. MC and GC immunoreactivity for the angiogenic factors is assumed to play an important role in the osteoclastogenesis process, thus contributing to additional growth of the CGCG lesions.     

1例同种异体骨片在唇侧骨板缺损位点即刻种植中的应用

目的 探讨使用同种异体骨片在唇侧骨板缺损位点进行不翻瓣即刻种植的临床效果，以期为该术式的临床应用提供参考。方法 对1例左上前牙残根伴慢性根尖周炎的病例进行不翻瓣即刻种植，应用隧道技术植入同种异体骨片重塑唇侧骨板，联合同期跳跃间隙内植骨和游离结缔组织移植完成软硬组织处理。结果 在种植体植入6个月后完成延期永久修复，CBCT显示植入同种异体骨片在位，间隙内成骨良好，种植体骨结合良好，完成个性化角度螺丝固位全瓷修复体安装。最终牙龈形态协调，PES评分为12分，获得了可接受的临床疗效。结论 同种异体骨片结合不翻瓣种植手术为唇侧骨板缺损位点的即刻种植提供了可预期的临床效果，扩大了即刻种植的临床适应证，减少了手术的创伤，有助于获得理想的临床疗效。     

Basic fibroblast growth factor and epidermal growth factor reverse impaired ulcer healing of the rabbit oral mucosa

BACKGROUND: The therapies for refractory ulcers on the oral mucosa are symptomatic and very unsatisfactory. We hypothesized that application of growth factors might be able to achieve successful remission of the lesion. We evaluated the effects of systemic administration and topical application of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on impaired wound healing of ulcers in the rabbit gingiva. METHODS: Almost uniform round ulcers could be created on the gingiva of the rabbits by chemical injury with acetic acid. When the submandibular glands were removed or i.v. injection of cisplatin (CDDP) and peplomycin sulfate was performed before ulcer formation, healing of the ulcers took longer than in untreated rabbits. To ascertain whether or not human EGF and bFGF affect rabbit cells, we first examined the effects of EGF and bFGF on the proliferation of the cells derived from rabbit gingiva. We then applied EGF or bFGF in these impaired healing models. RESULTS: EGF and bFGF promoted proliferation of the fibroblasts, and EGF also promoted proliferation of the keratinocytes isolated from gingival tissue of rabbits in vitro. Systemic injections of EGF and bFGF in rabbits, which had their submandibular glands removed, and topical application of bFGF accelerated healing of ulcers created in rabbits injected with CDDP and peplomycin sulfate. The ability of bFGF to promote the healing of ulcers was much greater than that of EGF. CONCLUSION: Basic FGF may be effective for refractory oral mucosal lesions.     

Enhanced bone regeneration around dental implant with bone morphogenetic protein 2 gene and vascular endothelial growth factor protein delivery

Objective: To evaluate the synergistic effect of bone morphogenetic protein 2 (BMP‐2) and vascular endothelial growth factor (VEGF) on the repair of bone defects around dental implants. Material and methods: Five groups of scaffold were fabricated by a freeze‐drying method, including pure chitosan/collagen scaffold; scaffold loaded with adenoviruses expressing BMP‐2, adenoviruses expressing VEGF, both adenoviruses expressing BMP‐2 and VEGF, VEGF protein and adenovirus expressing BMP‐2. In vitro studies examined whether bone marrow stromal cells were responsive to these scaffolds over time. Bone formation capacity, bone‐to‐implant contact, as well as removal torque values were investigated in vivo. Differences between the various groups were statistically analyzed using the one‐way analysis of variance test. Results: The in vitro study revealed a burst and rapid release of VEGF with a sustained high‐level expression of BMP‐2 in scaffold combined with VEGF protein and adenoviruses expressing BMP‐2. Histomorphometry demonstrated that scaffolds expressing BMP‐2 enhanced more bone formation compared with other groups; VEGF alone is insufficient to promote bone formation. New bone formation in the bone defects around dental implants, bone‐to‐implant contact and mean peak removal torque showed statistically significant difference for the adenoviral vector encoding human bone morphogenetic protein 2 (Ad‐BMP‐2) and VEGF protein and adenovirus expressing BMP‐2 groups. Furthermore, scaffold combined with VEGF protein and Ad‐BMP‐2 represented the best outcomes in this model. Conclusions: A combination of BMP‐2 gene and VEGF protein could have a synergistic effect in promoting bone healing. To cite this article:
Luo T, Zhang W, Shi B, Cheng X, Zhang Y. Enhanced bone regeneration around dental implant with bone morphogenetic protein 2 gene and vascular endothelial growth factor protein delivery.
Clin. Oral Impl. Res. 23 , 2012 467–474.
doi: 10.1111/j.1600‐0501.2011.02164.x     

婴幼儿脉管性疾病外周血和瘤内血bFGF的表达及意义

目的:通过检测婴幼儿血管性疾病外周静脉血(简称外周血)、瘤内血中的碱性成纤维细胞生长因子(bFGF)的表达,探讨对血管瘤和血管畸形进行鉴别诊断的可能.方法:应用酶联免疫吸附法(ELISA)检测资料完整的14例静脉畸形患儿瘤内血和外周血bFGF,检测49例血管畸形患儿、32例血管瘤患儿和23例对照婴幼儿外周血清中bFGF的浓度,采用SPSS11.5软件包对数据进行t检验和方差分析.结果:静脉畸形瘤内、外周血清中bFGF浓度有显著差异(P＜0.05),瘤内血清bFGF浓度高于外周血清:血管瘤、血管畸形和对照组外周血清bFGF浓度无统计学差异(P＞0.05).结论:静脉畸形患儿瘤内血清的bFGF浓度高于外周血清,检测外周血bFGF,不能鉴别血管瘤和血管畸形.     

bFGF对异种脱钙骨愈合过程中成骨细胞VEGF表达的影响

目的:了解不同浓度碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)与异种脱钙骨复合修复骨缺损时,成骨量和血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达水平之间的相关性。方法:在33只成年新西兰兔双侧下颌骨下缘形成15mm×5mm矩形骨缺损,每一缺损作为一个实验单位,术后4w取材作组织学检查,并分析成骨量和VEGF蛋白表达水平及其相互关系。结果:bFGF对于VEGF的表达与其诱导成骨作用一样存在双相剂量效应,VEGF表达和新骨形成呈密切关系。结论:bFGF可能是通过上调VEGF的表达,以促进血管化来实现增强异种脱钙骨愈合的。     

Time- and dose-dependent mitogenic effect of basic fibroblast growth factor combined with different bone graft materials: an in vitro study

OBJECTIVES: In periodontal regeneration, the growth factor concentrations and the delivery system used are of great importance. In an attempt to assess the mitogenic effect of basic fibroblast growth factor (bFGF) on periodontal ligament (PDL) cells combined with different bone replacement materials, two allografts of cortical (DFDBA) and cancellous (DFBA) bone and an anorganic bovine material with a synthetic peptide (ABM P-15) were used. The purpose of this study was to evaluate the in vitro mitogenic effect of different doses of bFGF alone or in combination with DFDBA, DFBA and ABM P-15 on human PDL cells in a time-dependent mode. MATERIAL AND METHODS: PDL cell cultures were derived from the mid-root of four maxillary premolars. Cells were grown and reached confluence. On day 2 of quiescence, new medium was added along with (1) 1, 5, 10 and 25 ng/ml of bFGF alone, (2) 10 mg of DFDBA, DFBA and ABM P-15 alone and (3) their combination. The mitogenic effect was determined at 24 and 48 h of culture by using a hemocytometer chamber. The cells were counted under a phase contrast microscope. RESULTS: The results revealed that bFGF at the highest concentrations and after 48 h exerted a significant mitogenic effect on PDL cells, and also DFDBA and DFBA supported cell proliferation. Furthermore, DFDBA and DFBA enriched with bFGF had a significant mitogenic effect after 48 h of culture. ABM P-15 with 10 and 25 ng/ml of bFGF up-regulated PDL cell proliferation after 48 h of incubation. CONCLUSIONS: The findings of this study demonstrate the beneficial role of bFGF combined with DFDBA and DFBA as carriers in periodontal repair.     

Temporospatial expression of vascular endothelial growth factor and basic fibroblast growth factor during mandibular distraction osteogenesis.

OBJECTIVE: Distraction osteogenesis is a vascular-dependent process. This study investigated expression patterns of two major angiogenic factors, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), in the distracted calluses following mandibular lengthening in a goat model. MATERIAL AND METHODS: Bilateral mandibular osteotomies were performed in 15 young adult goats. After a latency of 7 days, the mandibles were elongated using custom-made distractors with a rate of 1 mm/day for 10 days. Three animals each were sacrificed at the end of the delay phase, at 0, 7, 14, and 28 days after completion of distraction, respectively. The lengthened mandibles were harvested and processed for histological and immunohistochemical examinations. RESULTS: Elevated cellular expression of VEGF and bFGF, with neovascularization in the distraction gap, was observed following mandibular lengthening. VEGF staining was noted in the endothelial cells and osteoblasts. bFGF staining was seen in the fibroblast-like cells, osteoblasts and immature osteocytes. Their strongest expression was found 0-7 days after the end of distraction, and declined with maturation of the newly formed bone. CONCLUSION: A temporal and spatial expression pattern of VEGF and bFGF was found during distraction osteogenesis in goat mandibles. It suggests that distraction forces can stimulate the production of VEGF and bFGF, which contribute to neovascularization and new bone formation during gradual distraction of the mandible. Application of angiogenic factors may be considered as a potential method to enhance angiogenesis and osteogenesis in osteodistraction, especially in sites without enough vascularization.     

腺相关病毒介导VEGF促进新骨形成的实验研究

目的探讨腺相关病毒介导血管内皮生长因子165（VEGF165）在引导骨再生过程中对新骨形成的影响。方法体外构建承载VEGF基因的腺相关病毒。成年大耳白兔16只,随机分为对照组和实验组。在兔颅骨上建立引导骨再生模型,实验组充填血凝块及VEGF165腺相关病毒（rAAV-VEGF165）复合物,对照组充填血凝块。术后2周、4周,对充填部位进行组织学观察和组织形态测量学分析,比较实验组与对照组之间的异同。结果手术2周、4周后,实验组和对照组充填处均有新生骨和新生血管形成,实验组的新生骨高度和新生骨面积均显著高于对照组（P〈0.05）;实验组比对照组新生血管数量多,骨成熟度更高。结论 VEGF在引导骨组织再生中具有重要的作用,在引导骨再生早期应用rAAV-VEGF165可以有效地促进新骨形成。     

血清bFGF、E2及尿bFGF对婴幼儿血管瘤和脉管畸形的诊断意义

目的:探讨血清碱性成纤维细胞生长因子(bFGF)、雌二醇(E2)及尿bFGF在血管瘤和脉管畸形患者中的表达差异,为血管瘤和脉管畸形的鉴别诊断提供客观依据.方法:研究对象为78例增殖期血管瘤患者、18例消退期血管瘤患者及25例脉管畸形患者的血清和尿上清液,阴性对照为48例单纯唇腭裂患者,所有患者年龄均≤30个月.应用酶联免疫吸附分析法(ELISA)检测各组血清和尿bFGF浓度,并对尿bFGF浓度进行尿肌酐平衡;应用放射免疫方法(RlA)检测各组血清E2浓度.采用SPSS11.5软件包对数据进行统计学处理.结果:血管瘤患者血清bFGF浓度>脉管畸形患者血清bFGF浓度>对照组血清bFGF浓度,3组之间浓度有显著差异(P=0.027);血管瘤患者尿bFGF浓度>脉管畸形患者尿bFGF浓度>对照组尿bFGF浓度,3组之间浓度有显著差异(P=0.000);增殖期血管瘤患者尿bFGF浓度显著高于消退期血管瘤患者(P=0.04);血管瘤、脉管畸形患者和对照组3组之间血清E2浓度有显著差异(P=0.000);脉管畸形患者和对照组的血清E2浓度无显著差异(P>0.05).结论:血清、尿bFGF浓度可作为血管瘤鉴别诊断的指标,血清E2浓度可用于诊断血管瘤和脉管畸形并判断血管瘤是否处于增殖期.     

碱性成纤维细胞生长因子促进颌骨缺损修复的研究

目的 :评价碱性成纤维细胞生长因子 (bFGF)对成骨的促进作用 ,以期为颌骨缺损提供一种较为理想的替代材料。方法 :18只成年兔双侧下颌骨下缘各造成 15mm× 6mm的全层骨膜骨质缺损 ,每一缺损作为一个实验单位 ,按自身同期配对设计 ,分别植入复合骨和天然型无机骨 (NNB)。术后 3、6、12周取下标本 ,进行甲苯胺蓝染色观察、四环素荧光检查以及组织学成骨面积定量分析。结果 :复合骨新骨形成及钙化早 ,同期成骨面积大于NNB(P <0 .0 1)。结论 :合适的复合条件下bFGF可有效地促进软骨及骨的生长 ,复合骨新骨形成及钙化早 ,同期成骨量多 ,修复效果优于NNB。     

上颌快速扩弓腭中缝血管内皮生长因子的时空表达与新骨形成的相关性研究

目的 探讨上颌快速扩弓扩张期和固定期兔腭中缝组织血管内皮生长因子（VEGF）的时空表达模式以及新骨形成情况。方法 将44只新西兰大白兔随机分为11组：实验组（共5组）、对照组（共5组）、对照0组,每组4只。用螺旋分裂基托扩大矫治器（Haas矫正器）扩张兔上牙弓,快速扩张2周,固定4周。在安装扩张器当天（对照0组）、快速扩张第1、2周、固定第1、2、4周（实验组和对照组）取兔上颌骨腭中缝组织块,采用免疫组织化学法检测VEGF在腭中缝组织中的分布和表达变化,采用过碘酸-Schiff染色法观测新骨形成。结果 快速扩张的腭中缝可见高水平的VEGF表达,VEGF阳性信号主要定位于血管内皮细胞胞浆和增殖活跃的成骨细胞胞浆。对照组VEGF在整个实验过程中均呈弱阳性表达。实验组快速扩张第1周、快速扩张第2周、固定第1周、固定第2周VEGF蛋白表达量均高于对照组。随着机械应力快速扩张,实验组VEGF蛋白表达量逐渐上升,固定1周达峰值后逐渐下降。实验组新骨形成的量均高于对照组,新骨形成的量逐渐上升,固定2周达峰值后逐渐下降。结论 上颌快速扩弓产生的机械牵张力可以导致VEGF生成增加从而促进血管及新骨的生成。     

毛细血管瘤组织中成纤维细胞生长因子（bFGF）的表达水平

成纤维细胞生长因子（bFGF）是一组结构相关的多肽类血管形成因子，它能诱导或促进新生毛细血管的形成，对血管瘤的发展有关键作用。本文通过免疫组织化学方法，观察bFGF在颌面部毛细血管瘤不同分期的表达变化，初步探讨它在肿瘤增殖退化病理演变过程中的作用。     

大鼠正畸牙齿移动过程中牙周组织VEGF的表达

目的 :观察血管内皮生长因子 (vascularendothelialgrowthfactor ,VEGF )在大鼠正畸牙齿移动中牙周组织内的表达和分布 ,探讨VEGF在正畸牙齿移动中的作用机制。方法 :3 0只雄性SD大鼠 ,随机分为 6组 ,每组 5只 ,分 1、3、7、14、2 1d正畸加力组和正常对照组。采用免疫组织化学方法检测牙周组织VEGF的表达 ,并采用计算机图像分析方法对各组VEGF的表达强度进行半定量分析。结果 :VEGF在正畸加力组大鼠牙周组织中的表达明显强于正常对照组 ,加力 3d组的压力侧VEGF表达增强 ,其张力侧也有VEGF的表达 (略低于压力侧 ) ,7、14d为表达高峰期 (P <0 .0 1) ,2 1d组 ,VEGF表达有所减弱。VEGF的表达位于血管内皮细胞、成纤维细胞、成骨细胞和破骨细胞胞浆内。结论 :正畸牙齿移动中VEGF表达的变化 ,说明VEGF参与了牙齿移动过程中的牙周组织改建 ,可能对正畸牙齿移动修复具有重要意义。     

Microvessel density and vascular endothelial growth factor expression in sinus augmentation using Bio-Oss

AIM: The aim of this study was to evaluate microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression in sinus augmentation with Bio-Oss. METHODS: Twenty patients participated in this study. The sinuses were filled with 100% Bio-Oss. Implants were inserted after 3 months in group A, and 6 months in group B. A trephine was used to harvest bone cores. As control, the pre-existing subantral bone was used. RESULTS: The mean MVD in control bone was 23.6 +/- 1.8. In the sites augmented with Bio-Oss, at 3 months, the MVD was 23.3 +/- 2.1, while in the sites retrieved at 6 months the MVD was 29.5 +/- 2.4. The difference in MVD between the control bone and group A was not statistically significant. The difference between the control bone and group B was statistically significant (P < 0.05). The statistical analysis showed that the difference in MVD between group A and group B was statistically significant (P < 0.05). CONCLUSIONS: Bio-Oss seemed to induce an increase in MVD that reached a higher value after 6 months. The percentage of vessels positive to VEGF was higher in group A than in group B. Our data also showed a higher percentage of vessel and stromal cells positive to VEGF and higher MVD values in areas where there was newly formed bone compared with areas where maturation processes were occuring, and this fact could point to a close spatial relationship between angiogenesis and osteogenesis.