基质金属蛋白酶-8和基质金属蛋白酶-13在人根尖周病组织肥大细胞上的表达

目的:观察基质金属蛋白酶-8(matrixmetalloproteinase-8,MMP-8)和基质金属蛋白酶-13(matrixmetalloproteinase-13,MMP-13)在人慢性根尖周炎组织肥大细胞(mastcells,MCs)上的表达情况,探讨肥大细胞在根尖周病发病机制中的作用。方法:将90例样本分为3组:1)根尖肉芽肿组30例;2)根尖囊肿组30例;3)正常牙周膜组30例。组织标本置于10%中性福尔马林固定液中浸泡48h以上,制作连续组织切片。HE染色,光学显微镜下观察各组的组织学变化;免疫荧光双染色(doubleimmunofluorescence,DIF),荧光显微镜下观察MMP-8和MMP-13在根尖周组织MCs上表达的情况。结果:与正常对照组相比,两组慢性根尖周炎组织中MMP-8和MMP-13双阳性MCs数目均显著增多(P<0.01);根尖囊肿组与根尖肉芽肿组MMP-8和MMP-13双阳性MCs密度无显著性差异。结论:慢性根尖周炎病变组织中MCs数目明显增多,同时MMP-8和MMP-13双阳性MCs密度显著增高,提示MMP-8和MMP-13阳性MCs可能在慢性根尖周炎的致病机制中发挥作用。     

基质金属蛋白酶在牙髓炎症中的活性及表达

目的探讨来源于宿主的基质金属蛋白酶在牙髓炎发病机制中的作用。方法利用免疫组化和明胶酶谱法,检测健康和炎症牙髓中MMP-2、MMP-9的表达定位及酶活性。结果MMP-2、MMP-9在正常和炎症牙髓中均有表达,定位于成牙本质细胞及牙髓成纤维细胞,炎症浸润区表达强阳性。明胶酶谱分析,MMP-9酶原在正常和炎症牙髓均被检测到,且炎症组表达高于正常组。结论MMP-9可能参与牙髓组织破坏过程,在炎症牙髓组织降解过程中起重要作用。     

口腔癌相关成纤维细胞对人工基底膜的降解作用及其机制

目的观察口腔癌相关成纤维细胞（CAFs）降解人工基底膜matrigel的能力,并探讨其可能的作用机制。方法matrigel凝胶与CAFs和NFs共同培养24、48、72 h后,收集培养上清液,按照羟脯氨酸检测试剂盒说明测定羟脯氨酸含量;用考马斯亮兰微孔板法行微量蛋白定量;用明胶酶谱分析对基质金属蛋白酶- 2（MMP- 2）、基质金属蛋白酶- 9（MMP- 9）含量进行测定。结果在各时间段,口腔CAFs上清超滤液中羟脯氨酸含量、上清液总蛋白浓度及MMP- 2酶原含量明显高于NFs,且CAFs组表达大量MMP- 2活性酶。口腔CAFs和NFs皆不表达MMP- 9。结论CAFs具有较强的降解细胞外基质的能力,提示其与肿瘤- 宿主微环境中基质重建及肿瘤侵袭有关。     

基质金属蛋白酶及其组织抑制剂表达失衡与涎腺恶性肿瘤浸润生长的关系

目的　探讨基质金属蛋白酶2、9(MMP-2、9),膜型基质金属蛋白酶1(MT1-MMP)及基质金属蛋白酶组织抑 制剂1、2(TIMP-1、2)与涎腺恶性肿瘤浸润生长的关系。方法　应用免疫组化SP法和明胶酶谱法检测28例涎腺良 性肿瘤、26例涎腺恶性肿瘤中MMP-2、9,MT1-MMP及TIMP-1、2的表达及细胞定位,分析其中酶原与活性酶的含量 比例。结果　涎腺恶性肿瘤中MMP-2、9的表达强于良性肿瘤(P<0·05),TIMP-1、2的表达低于良性肿瘤 (P<0·05)。MMP-2、MT1-MMP、TIMP-2的表达有相关性。MMP-9酶原及活性MMP-9在恶性肿瘤中的表达均高于良 性肿瘤(P<0·05)。结论　MMP-2、9在涎腺恶性肿瘤浸润性生长中扮演了重要角色。涎腺恶性肿瘤中,TIMP-1、2 抑制作用的降低导致了MMP-2、9合成、激活的增多,令MMP-2、9与其天然组织抑制剂TIMP-1、2之间的平衡失调, 从而基底膜加速降解。     

基质金属蛋白酶诱导因子和亲环素A在人根尖囊肿和根尖肉芽肿中的表达及临床意义

目的: 检测细胞外基质金属蛋白酶诱导因子(CD147)和亲环素A(CypA)在根尖肉芽肿和根尖囊肿中的表达,探讨CD147和CyPA在人慢性根尖周炎发生发展中的作用。方法: 收集经根尖手术切除所获得的根尖肉芽肿35例和根尖囊肿30例作为实验组,同时收集埋伏阻生智齿拔除术或行牙槽骨修整术凿下的8例健康牙槽骨作为正常对照组。CBCT图像记录病例的病损大小。运用免疫组织化学法检测所有样本中CD147和CypA的蛋白表达,分析CD147和CypA的蛋白表达水平。结果: 根尖囊肿组两种蛋白的表达水平均高于肉芽肿组(P<0.05)。CD147和CypA 的蛋白表达水平在根尖囊肿和根尖肉芽肿中呈正相关(P<0.05);CD147和CypA 的表达水平均与慢性根尖周炎的病变大小呈正相关(P<0.05)。结论: CD147和CypA可能参与根尖周病损的炎性反应和骨质吸收。CD147-CypA相互作用在人慢性根尖周病的发生发展过程中可能发挥了某种协同作用。     

核因子кB受体激动子配体在慢性根尖周炎病损组织中的表达

目的:探讨核因子кB受体激动子配体(receptor activator of nuclear factorκB ligand,RANKL)在慢性根尖周炎发病中的作用。方法:采用RT-PCR法,从mRNA水平检测RANKL在10例慢性根尖肉芽肿及3例根尖囊肿中的表达,分析RANKLmRNA的表达与慢性根尖周炎病损的病理学表现(包括病损的病理分型、炎症细胞浸润程度)之间的关系,采用免疫组化的方法,对RANKL的表达进行细胞定位。结果:RANKLmRNA在根尖肉芽肿病损组和根尖囊肿组明显增高,但根尖肉芽肿和根尖囊肿组之间差异无显著性。RANKL mRNA在轻、重度炎症细胞浸润组较中度炎症细胞浸润组明显增高(P<0.05)。RANKL mRNA水平与病损大小无关。免疫组化结果显示,RANKL主要表达于浸润的炎症细胞中及增生的上皮细胞中。结论:RANKL在慢性根尖周炎的发病中具有重要的作用,但也提示存在其它细胞因子的协同作用。     

核因子κB受体激动子配体在慢性根尖周炎病损组织中的表达

目的:探讨核因子κB受体激动子配体(receptor activator of nuclear factor κB liganol,RANKL)在慢性根尖周炎发病中的作用.方法:采用RT-PCR法,从mRNA水平检测RANKL在10例慢性根尖肉芽肿及3例根尖囊肿中的表达,分析RANKLmRNA的表达与慢性根尖周炎病损的病理学表现(包括病损的病理分型、炎症细胞浸润程度)之间的关系,采用免疫组化的方法,对RANKL的表达进行细胞定位.结果:RANKLmRNA在根尖肉芽肿病损组和根尖囊肿组明显增高,但根尖肉芽肿和根尖囊肿组之间差异无显著性.RANKLmRNA在轻、重度炎症细胞浸润组较中度炎症细胞浸润组明显增高(P＜0.05).RANKL mRNA水平与病损大小无关.免疫组化结果显示,RANKL主要表达于浸润的炎症细胞中及增生的上皮细胞中.结论:RANKL在慢性根尖周炎的发病中具有重要的作用,但也提示存在其它细胞因子的协同作用.     

基质金属蛋白酶与根尖周病

基质金属蛋白酶（matrix metalloproteinases,MMPs）属于锌依赖性蛋白水解酶的一种,是由多种蛋白酶组成的多功能内肽酶,主要降解细胞外基质（extracellularmatrix,ECM）。在正常稳定状态组织中MMP表达量极少,而在炎性细胞因子、激素、生长因子刺激下和细胞转化过程其表达上升。病理上根尖周病通常分为根尖周炎和根尖囊肿,根尖周炎多是由根管内微生物感染引起的局部炎症性疾病,常累及邻近牙槽骨和根尖部的牙骨质,使其吸收破坏。     

基质金属蛋白酶及其组织抑制剂在牙本质生成性影细胞瘤和牙源性影细胞癌中的表达

目的 对牙本质生成性影细胞瘤(dentinogenic ghost cell tumor,DGCT)和牙源性影细胞癌(ghost cell odontogenic carcinoma,GCOC)中基质金属蛋白酶(matrix metallopmteinases,MMP)和金属蛋白酶组织抑制剂(tissue inhibitors of metalloproteinase,TIMP)的表达特点进行研究.方法 免疫组织化学分析15例DGCT和9例GCOC中MMP-2、MMP-9、MMP-14、TIMP-1和TIMP-2的表达.对DGCT和GCOC各一例分析MMP-2、MMP-9、MMP-14、TIMP-1和TIMP-2 mRNA的表达;明胶酶谱分析MMP-2和MMP-9酶原和活化蛋白的表达.结果 MMP-9和TIMP-1在GCOC中高表达(7/9,8/9);TIMP-1在GCOC中的高表达,与在DGCT中相比差异具有统计学意义(P＜0.05).对一例DGCT和一例GCOC病例研究发现,MMP-9酶原和活酶在GCOC中高表达,并且MMP-9和TIMP-1 mRNA在GCOC中高表达.结论 MMP-9和TIMP-1可能与GCOC的性质和生物学行为有关.     

干细胞因子在人慢性根尖周炎组织中表达的研究

目的:观察干细胞因子(stem cell factor, SCF)在人慢性根尖周炎病变组织中成纤维细胞(fibroblasts, FBs)、内皮细胞(endothelial cells, ECs)及巨噬细胞(macrophages, m )上的表达,探讨SCF阳性的FBs、ECs及m 在人慢性根尖周炎发病机制中的作用。方法:本实验共收集50例根尖周组织标本,其中根尖周肉芽肿组15例,根尖周囊肿组15例,正常对照组20例。组织标本经福尔马林液固定,石蜡包埋、制作连续切片。HE染色,光学显微镜下观察各组标本的组织学变化;经免疫荧光双染色(double immunofluorescence, DIF),在荧光显微镜下观察SCF在根尖周组织FBs、ECs及m 上表达的情况。结果:DIF结果显示,两组慢性根尖周炎组织中的CD334-SCF双阳性FBs、CD31-SCF双阳性ECs及CD14-SCF双阳性m 密度较正常对照组显著增多(P＜0.01);根尖周囊肿组与根尖周肉芽肿组的CD334-SCF双阳性FBs及CD31-SCF双阳性ECs密度无显著性差异;根尖周肉芽肿组CD14-SCF双阳性m 密度明显高于根尖周囊肿组(P＜0.01)。结论:结果提示CD334-SCF双阳性FBs、CD31-SCF双阳性ECs及CD14-SCF双阳性m 与慢性根尖周炎的发生、发展及致病机制有关。     

大鼠牙移动过程中牙周组织MMP-2、9/TIMP-2水平变化的研究

目的 观察大鼠牙正畸移动过程中MMP-2、9及其抑制剂TIMP-2在牙周组织中的表达和分布。方法 ASD大鼠30只,分为空白对照组、牙齿移动1、2、3、和4周组。加力将上颌第一磨牙向近中移动,于1、2、3、4周将动物处死,制作石蜡包埋切片,用免疫组化染色观察并比较各组中第一磨牙牙周组织中MMP-2、9及其抑制剂TIMP-2的表达和分布。进行统计学分析。结果 MMP-2、9在受力牙齿压力侧牙周组织的破骨细胞和张力侧牙周组织的成骨细胞以及成纤维细胞中的阳性表达上调,并随受力时间变化而波动。TIMP-2随着时间变化逐渐轻度上升。结论 牙齿移动过程中MMP-2、9上调可造成MMPs/TIMPs的失衡,并且这种上调可能参与了正畸牙齿移动过程中牙周组织的改建。?     

Matrix metalloproteinase-8 (MMP-8) in pulpal and periapical inflammation and periapical root-canal exudates

AIM: To study the presence, levels and molecular forms of matrix metalloproteinase (MMP) -8 (collage-nase-2) in pulpal and periapical inflammation, and the changes in MMP-8 levels in root-canal exudates during root-canal treatment. METHODOLOGY: Periapical exudate samples were collected from 11 necrotic teeth with radiographically verified periapical periodontitis during three root-canal treatment visits with interappointment calcium hydroxide (Ca(OH)2) medication. MMP-8 levels and molecular forms were analyzed with immunofluorescent assay (IFMA) and Western immunoblot. Inflamed pulp tissue and periapical granuloma tissue (n = 10 for both) were obtained from other patients and used for MMP-8 immunohistochemical (IHC) staining. RESULTS: The periapical exudate samples demonstrated marked differences in MMP-8 levels between the teeth in the first visit and significant decrease in MMP-8 levels during the root-canal treatment (P = 0.0107). One specimen failed to show a decrease in MMP-8 below 1000 ng mL(-1) a vertical root fracture was later diagnosed and the tooth extracted. IHC staining showed that in addition to PMN-leucocytes, macrophages and plasma cells produced MMP-8 in pulp and periapical granulomas. CONCLUSIONS: The findings demonstrate the presence of MMP-8 in the inflamed pulp and periapical tissue, indicating that MMP-8 has a role in pulpal and periapical inflammation, most likely participating in tissue extracellular matrix degradation. They further indicate that MMP analysis from periapical exudate could be used to indicate and monitor inflammatory activity and the success of treatment in teeth with periapical lesions.     

Immunolocalization of matrix metalloproteinases-2 and -9 during apical periodontitis development

Objective

The objective of this study was to determine the expression of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) during apical periodontitis development.

Methods

Using an experimental design of induced periapical lesions in rats and immunohistochemistry assay as investigative tool, the MMP-2 and MMP-9 expression and distribution were evaluated at 3, 7, 14, 21, 30, 60 and 90 days after coronary access and pulp exposure of the first left mandibular molar to the oral environment. Two blind observers scored the immunoreactivity. A semi-quantitative analysis was performed.

Results

Except at day 3, MMP-2 and MMP-9 immunostaining was observed in all experimental periods. The MMP-2 (p = 0.004) and MMP-9 (p = 0.005) immunostaining was higher in the period between 7 and 21 days. They were mainly observed in cells surrounding the apical foramen and adjacent periapical areas. Cells into the hypercementosis areas were strongly stained while both osteoblasts and osteoclasts presented discrete staining along of this study. No staining was observed on epithelial walls. At 30, 60 and 90 days, the subjacent connective tissue presented intense MMP-2 and MMP-9 immunostaining in mononuclear cells (suggestive of fibroblasts, macrophages, infiltrating neutrophils and lymphocytes).

Conclusion

The results observed in this study suggest that MMP-2 and MMP-9 play a critical role in the development of inflammatory periapical lesions, probably involved in the extracellular matrix (ECM) degradation during the initial phase of the lesion development.     

Association of matrix metalloproteinase inducer (EMMPRIN) with the expression of matrix metalloproteinases-1, -2 and -9 during periapical lesion development

Objective

To evaluate the expression of matrix metalloproteinase inducer (EMMPRIN) and its correlation with the expression of matrix metalloproteinases (MMPs)-1, -2 and -9 during the development of periapical lesion in mice.

Methods

Periapical lesions were induced in the lower first molars of mice and after 7, 14, 21 and 42 days the mandibles were removed. The periapical lesions were measured by micro-computed tomography. The expression of EMMPRIN, MMPs-1, -2, and -9 genes were determined by real-time RT-PCR. The location and expression of EMMPRIN and MMPs were evaluated by immunohistochemistry.

Results

At 14 days, the periapical lesion area was higher than at 7 days. At 21 and 42 days no statistically significant bone loss was observed in comparison to 14 days. The control group showed discrete and occasional EMMPRIM, MMP-1, -2 and -9 immunostaining in the periodontal ligament fibroblasts. At 7, 14, 21 and 42 days intense immunoexpression was observed for EMMPRIN, MMPs-1, -2 and -9 in the region adjacent to the apical foramen. The EMMPRIN immunoexpression was higher at 7, 14, 21 and 42 days compared with the control. There was a positive correlation between gene expression of EMMPRIN and MMPs in the active phase of periapical lesion development.

Conclusion

There is a high expression of EMMPRIM mainly by the inflammatory infiltrate in the region adjacent to the apical foramen during periapical lesion development. Furthermore, the positive correlation with MMP-1, -2, and -9 during the first days after periapical lesion induction indicates that EMMPRIM may be involved in the active phase of periapical lesions development.     

Collagenase-3 (MMP-13) is expressed in periapical lesions: an immunohistochemical study

AIM: To determine whether or not matrix metalloproteinase 13 (MMP-13) is present in periapical granulomas with and without epithelium. METHODOLOGY: Seventeen open periapical granulomas of pulpal origin (seven lesions without epithelium and 10 with proliferating epithelium) were fixed in formalin and then embedded in paraffin prior to being processed for immunohistochemical analysis. A monoclonal antibody against human MMP-13 was used to evaluate MMP-13 expression. Immunocomplexes were subsequently treated with the secondary antibody and then detected by means of streptavidin peroxidase. Immunoreactivity was visualized by development with 3,3'-diaminobenzidine. RESULTS: An immunopositive cytoplasmatic reaction for MMP-13 was observed in all the specimens, although the immunostaining by anti-MMP-13 antibody was heterogeneous and its levels varied according to histopathological findings. In periapical lesions without epithelium MMP-13 immunolabelling was detected in a few fibroblast-like cells, and in some plasma cells within the granulomatous tissue. A clear upregulation of MMP-13 expression was detected in periapical lesions with epithelium, especially in small island and thin strands of epithelium. CONCLUSIONS: The expression pattern of MMP-13 demonstrates that it is involved in the conversion of a periapical granuloma with epithelium into a radicular cyst. This property is related to the ability of MMP-13 to influence not only the migration of epithelial cell but also the invasion of granulomatous tissue.     

Healing of extraction sockets in collagenase-2 (matrix metalloproteinase-8)-deficient mice

Matrix metalloproteinase-8 (MMP-8) participates in skin wound healing and inflammation. We hypothesized that MMP-8 plays a role in wound healing after tooth extraction and in periapical inflammation. Bone formation, collagen metabolism, and inflammation in tooth extraction socket and in periapical lesions were analyzed in wild-type mice and in MMP-8-deficient (MMP-8^−/−) mice. New trabecular bone area in the extraction sockets and in periapical lesions were similar in both groups. In extraction sockets significantly more type III procollagen was synthesized, and the neutrophil and MMP-9 levels were lower in MMP-8^−/− mice. The amount of Fas ligand, identified as a substrate for MMP-8, was lower in alveolar mucosa but higher in alveolar bone of MMP-8^−/− mice. These results indicate that MMP-8 can modulate inflammation and collagen metabolism of alveolar bone and mucosa.     

胶原膜与羟基磷灰石修复牙槽骨缺损的远期疗效观察

目的：观察国产BME-10X型医用胶原膜与羟基磷灰石(HA)联合引导组织再生治疗牙周炎、根尖囊肿造成骨缺损的I临床疗效。方法：选择18例患者的9颗Ⅱ度根分叉病变牙的9个位点、11颗近远中二、三壁骨下袋牙的16个位点、6颗根尖囊肿牙，在骨缺损区填入HA后，用胶原膜覆盖，观察1年以上的愈合情况。结果：骨下袋病变成功率75％，根分叉病变成功率77.8％，根尖囊肿成功率100％。结论：胶原膜／HA引导再生术在牙槽骨缺损修复中有一定应用价值。