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.     

Basic fibroblast growth factor up-regulates the expression of vascular endothelial growth factor during healing of allogeneic bone graft

Recently we reported that basic fibroblast growth factor (bFGF) improved the healing of allogeneic bone grafts. However, the mechanism of action of the bFGF was not known. Therefore, the present study was designed to identify the expression pattern of vascular endothelial growth factor (VEGF) in the presence of bFGF reconstituted in demineralized intramembranous bone matrix (DBM_IM) during the healing of allogeneic bone grafts. Eighteen critical size (15 mm × 10 mm) defects were created on rabbit mandibles bilaterally. Three groups of six defects each were grafted with allogeneic bone alone, allogeneic bone and DBM_IM, and allogeneic bone and bFGF reconstituted in DBM_IM. Three weeks later, the defects were retrieved for immunohistochemistry and in situ hybridization for VEGF. The percentage of positive staining area was quantified by using image analyzer. The increase (517%) in the expression of VEGF mRNA was accompanied by an increase (492%) of immunoreactive VEGF protein in allogeneic bone graft augmented by bFGF reconstituted in DBM_IM. A close correlation existed between levels of VEGF production and the amount of newly formed bone. The results show that bFGF reconstituted in DBM_IM markedly up-regulated the expression of VEGF in the grafted area. Basic FGF augments the healing of allogeneic bone grafts by enhancing vascularization through the up-regulation of VEGF.     

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.     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10-¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodiflerentiation of PDL cells into mineralized tissue forming cells.     

Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells

Basic fibroblast growth factor (FGF-2; bFGF) is a major mitogen for connective tissue cells, and participates in the healing process. It has already been reported that FGF-2 could be applicable to enhance periodontal regeneration. In the present study, we examined FGF receptor (FGFR) expression on human periodontal ligament (PDL) cells. The binding of [¹²⁵I]-labeled FGF-2 to human PDL cells was studied by radioreceptor assay. The binding of [¹²⁵I]-FGF-2 to PDL cells reached a plateau after 2.5 h incubation at 4°C and was inhibited by the addition of unlabeled FGF-2 and acidic FGF (FGF-1; aFGF), but not insulin-like growth factor-I, platelet-derived growth factor and transforming growth factor-β₁. Scatchard analysis revealed the presence of approximately 1.0 × 10⁵ FGF-2 binding sites per cell with an apparent Kd of 1.2 × 10⁻¹⁰ M. Interestingly, the binding of [¹²⁵I]-FGF-2 on PDL cells reached its maximum at d 6 of the culture and then gradually decreased. Scatchard analysis also demonstrated that the number of FGFRs on a PDL cell was altered during the course of the culture, while the affinity between FGF-2 and its receptor was not. The responsiveness of PDL cells to FGF-2, which was monitored by the inhibitory effect on alkaline phosphatase activity, was reduced in proportion to the decrease in the number of FGFRs on the PDL cells. The present study suggests that PDL cells alter the responsiveness to FGF-2 during the course of the culture by changing the density of its receptor, and that the density of FGFR expression might be a marker of the cytodifferentiation of PDL cells into mineralized tissue forming cells.     

碱性成纤维细胞生长因子在人牙胚中表达的免疫组化定位

目的：观察碱性成纤维细胞生长因子（ｂＦＧＦ）表达与人牙胚发育和分化的关系。方法：采用免疫组化方法观察ｂＦＧＦ在人牙胚发育中的定位。结果：钟状期牙胚中前成釉细胞为ｂＦＧＦ强阳性，星网层可疑；成牙本质细胞为强阳性，牙乳头组织中阳性分布不均，靠近成牙本质细胞的牙乳头细胞为阳性；牙囊细胞阳性，血管内皮细胞呈强阳性；骨组织为阴性。结论：ｂＦＧＦ与成釉细胞、成牙本质细胞的分化和成熟有关。     

NGF、bFGF对体外培养人牙髓细胞增殖与分化的作用

目的:研究NGF、bFGF及两者联合应用对体外培养人牙髓细胞(HDPC)的增殖与分化作用的影响。方法:用MTT和ALP活性检测法,观察对照组(10ml/L FBS的DMEM培养液)与实验组(10U/ml NGF、10μg/L bFGF、10U/ml NGF 10μg/L bFGF、5U/ml NGF 5μg/L bFGF、1U/ml NGF 1μg/L bFGF)对体外培养的第5~8代HDPC增殖与分化作用的影响。对实验数据行Dunnett-t检验。结果:与对照组相比,10U/ml的NGF可显著促进HDPC的增殖(P<0.05);对ALP的活性没有明显的增强作用(P>0.05);10μg/LbFGF可显著促进HDPC的增殖(P<0.05),但对HDPC的ALP活性,却有一定的抑制作用(P>0.05);10U/ml NGF 10μg/L bFGF和5U/ml NGF 5μg/L bFGF既可显著地促进HDPC增殖(P<0.05),又可增强HDPC的ALP活性(P<0.01)。结论:一定浓度的NGF与bFGF组合既可促进HDPC增殖,又可促进其分化,两者对HDPC有显著的功能放大性协同增强作用。     

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。     

乳铁蛋白对口腔癌细胞血管内皮生长因子和碱性成纤维细胞生长因子表达的影响

目的 观察乳铁蛋白对口腔癌细胞株Tca8113细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)表达的影响.方法 实验分0.006、0.013、0.025、0.050 g/L乳铁蛋白实验组和空白对照组.反转录聚合酶链反应(RT-PCR)和蛋白质印迹法分别检测VEGF、bFGFmRNA和蛋白表达的改变.结果 0.050 g/L乳铁蛋白实验组VEGFmRNA的表达(0.31±0.08)和蛋白的表达(0.68±0.11)均显著低于空白对照组[分别为(1.07±0.13)、(0.97±0.07)],P<0.05;0.050 g/L乳铁蛋白实验组bFGFmRNA的表达(0.27±0.10)和蛋白的表达(0.68±0.07)均显著低于空白对照组[分别为(0.91±0.12)、(1.01±0.09)],P<0.05.结论 乳铁蛋白能抑制口腔癌细胞株Tca8113细胞中VEGF、bFGF的转录和表达,抑制肿瘤的血管发生可能是乳铁蛋白抗口腔癌的机制之一.     

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.     

碱性成纤维细胞生长因子对牙周膜细胞表皮生长因子受体基因表达的影响

目的研究碱性成纤维细胞生长因子（bFGF）对人牙周膜细胞（PDLC）表达表皮生长因子受体（EGFR）的影响,探讨bFGF在牙周组织分化再生中的意义。方法体外原代培养人PDLC,有限稀释法形成单细胞克隆,用外源性bFGF刺激单细胞克隆,采用逆转录聚合酶链反应（RT-PCR）检测克隆细胞内EGFR基因表达的变化。结果bFGF促进人PDLC内EGFR mRNA的合成,并且随着质量浓度的增加促进作用增强。结论bFGF对EGFR的促进作用很可能是牙周炎损伤修复过程中一个重要的调节因素,为牙周组织分化再生提供部分理论基础。     

bFGF对人牙髓、牙周膜成纤维细胞生物学效应的研究

目的：了解碱性成纤维细胞生长因子（bFGF）对人牙髓、牙周膜成纤维细胞的生物学效应,为bFGF在牙髓、牙周病中的治疗研究提供新的实验依据。方法：利用3H-TdR掺入法观察在bFGF作用下人牙髓、牙周膜成纤维细胞DNA和胶原蛋白合成的情况。结果：20ng/mL～60ng/mLbFGF可明显促进人牙髓、牙周膜成纤维细胞DNA的合成（P〈0.01）,在40ng/mL浓度时牙髓,牙周膜成纤维细胞DNA合成最高,40ng/mLbFGF作用于牙髓,牙周膜成纤维细胞,24～48h可使细胞DAN合成显著增多,牙髓成纤维细胞在36h时DNA合成达最高峰,牙周膜成纤维细胞在24h时DNA合成达最高峰;bFGF对牙髓,牙周膜成纤维细胞胶原蛋白的合成无明显促进作用（P〉0.05）。结论：人牙髓、牙周膜成纤维细胞是bFGF的靶细胞,其胞膜上可能有bFGF特异性受体的存在,也表明bFGF在牙髓、牙周组织的创伤愈合中可能起重要作用。     

bFGF和rhBMP对牙周膜细胞胶原酶水平的影响

目的:研究bFGF、rhBMP分别及联合作用对人牙周膜细胞(periodontalligamentcells ,PDLCs)胶原酶Ⅱ、Ⅳ(CollagenaseⅡ、Ⅳ)水平的影响并探讨其内在相关性;优选最佳显效浓度。方法:选取因正畸拔除的第一前磨牙,刮取根中1/3牙周膜组织作为标本,采用牙周膜细胞体外培养技术,免疫组化方法和图像分析技术动态观察bFGF、rhBMP梯度含量分别及联合作用对人PDLCs胶原酶Ⅱ、Ⅳ水平的影响。结果:①免疫组化定量分析,bFGF(10 )浓度组能显著抑制PDLCs胶原酶Ⅳ的表达(P <0 .0 1) ;rhBMP(2 0 0 )浓度组能诱导PDLCs胶原酶Ⅳ的表达(P<0 .0 5 ) ;bFGF(10 )、rhBMP(2 5 )浓度组均能显著诱导PDLCs胶原酶Ⅱ的表达(P <0 .0 1) ;动态观察bFGF(10 ) rhBMP(2 0 0 )连续7d对胶原酶Ⅱ、Ⅳ水平的抑制效应呈现连续的递增趋势(P <0 .0 5 )。②rhBMP与人PDLCs胶原酶Ⅱ水平呈显著负相关性(P <0 .0 1) ;rhBMP与人PDLCs胶原酶Ⅳ水平呈显著正相关性(P <0 .0 1)。结论:bFGF、rhBMP联合作用能更显著地抑制人PDLCs胶原酶Ⅱ、Ⅳ的表达,两者具有协同作用;其中bFGF(10 ) rhBMP(2 0 0 )为最佳显效浓度。     

动态观察bFGF和rhBMP对牙周膜细胞增殖的影响

目的 探讨碱性成纤维细胞生长因子(bFGF)和重组人骨形成蛋白(rhBMP)分别及联合作用对人牙周膜细胞(PDLCs)增殖的影响；优选其最佳显效浓度。方法 选取因正畸治疗需要拔除的第一前磨牙，刮取根中1／3牙周膜组织作为标本，采用牙周膜细胞的体外培养技术，用四唑盐比色法(MTT)和酶动力学方法，动态观察bFGF、rhBMP梯度含量分别及联合作用对人PDLCs增殖的影响。结果 bFGF、rhBMP分别作用均能促进PDLCs的增殖；bFGF(10) rhBMP(200)(最佳浓度之和)促PDLCs增殖作用均较其分别作用更加显著；动态观察bFGF(10) rhBMP(200)作用于PDLCs 1～7d，显示出连续递增的促增殖趋势。结论 bFGF、rhBMP分别作用均能促进人PDLCs的增殖，联合应用则更具有协同作用；其中bFGF(10) rhBMP(200)为最佳显效浓度     

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.     

PTTG和bFGF在口腔鳞癌中的表达及意义

目的：探讨垂体肿瘤转化基因（pituitary tumor transforming gene，PTFG）和碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）在口腔鳞癌中的表达及相互关系，研究它们的表达与肿瘤临床病理指标的联系。方法：应用SP染色法检测PTTG蛋白和bFGF在55例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率。结果：在口腔鳞癌中PTTG和bFGF的阳性表达率分别为78．2％和67．3％，其阳性率及表达等级均显著高于正常对照组（P〈0．05）。PTTG在中一低分化组和有淋巴结转移组中的表达显著高于高分化组和无淋巴结转移组（P〈0．05）。PTTG表达与bFGF表达成等级正相关（r=0．382，P〈0．05）。结论：PTFG或bFGF与口腔鳞癌生物学行为及预后有密切关系，二者的联合检测，有助于口腔鳞癌恶性程度和预后的判断。     

Microvessel density and expression of vascular endothelial growth factor, basic fibroblast growth factor, and platelet-derived endothelial growth factor in oral squamous cell carcinomas

Angiogenesis, the growth of capillary vessels, plays an important role in the metabolic functions of malignant tissues. Tumor growth and malignant transformation are considered to be dominated by uncontrolled angiogenesis. To understand the mechanism of increased vascularity associated with malignant tissues, we immunohistochemically evaluated microvessel density (MVD) and the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and platelet-derived endothelial growth factor (PDGF) in oral cancers. Microvessel density did not differ significantly between normal oral mucosa and epithelial dysplasia, but was significantly increased in tumor tissues. Expression of angiogenic factors was not found in normal oral mucosa, but increased in association with increasing vascularity in OSCC tissue. In tumor tissue, angiogenic factor expression correlated with MVD. MVD in OSCC was related to T stage, tumor differentiation, and stage of invasion. VEGF expression also correlated with tumor differentiation and the stage of invasion. These findings suggest that VEGF might play an important role in tumor angiogenesis of OSCC.