尼古丁对大鼠不同部位骨钙磷含量和碱性磷酸酶活性的差异影响

目的 分析尼古丁对大鼠牙槽骨和颌骨内钙磷含量以及碱性磷酸酶（ALP）活性的影响。方法 将20只5周龄健康雄性Wistar大鼠随机分为实验组和对照组,实验组每天腹腔注射尼古丁0.73 mg•kg-1,对照组腹腔注射等量生理盐水,3个月后处死取材。浓酸消化法检测牙槽骨和颌骨内钙磷含量,改良Reddi法检测牙槽骨和颌骨内ALP活性。采用SPSS 16.0统计软件分别行独立样本的t检验。结果 与对照组相比较,实验组牙槽骨内和颌骨内钙磷含量均明显减少,差异均有统计学意义（P<0.05）;ALP活性无明显变化（P>0.05）。实验组中牙槽骨与颌骨比较,骨钙磷含量和ALP活性差异无统计学意义（P>0.05）。结论 尼古丁下调大鼠牙槽骨和颌骨内钙磷含量,对ALP活性的影响不明显。     

被动吸烟对大鼠牙槽骨内DSP和OPN表达的影响

目的:观察被动吸烟对大鼠牙槽骨内牙本质涎蛋白(dentin sialoprotein,DSP)及骨桥蛋白(osteopontin,OPN)表达的影响。方法:20只6周龄健康雄性Wistar大鼠随机等分为对照组和实验组,每个大组再等分为30d和60d两个亚组。采用单纯烟熏法建立大鼠被动吸烟模型,免疫组织化学法检测DSP和OPN在大鼠牙槽骨内的表达。结果:与对照组大鼠相比较,实验组大鼠牙槽骨内DSP和OPN表达均明显上调;在本实验周期内,上述变化随实验时间的延长而加重。结论:被动吸烟上调大鼠牙槽骨内矿化相关非胶原蛋白DSP及OPN的表达,推测DSP和OPN在被动吸烟抑制牙槽骨矿化的发病机制中发挥一定的作用。     

高脂饮食对大鼠下颌骨钙、磷及羟脯氨酸含量以及对牙槽骨骨量的影响

目的:探讨高脂饮食与大鼠下颌骨钙、磷及羟脯氨酸含量以及牙槽骨骨量的关系。方法:将20只SD大鼠随机分为正常对照组和脂肪乳剂组,正常对照组予普通饲料及生理盐1mL/100g体重灌胃,脂肪乳剂组予普通饮食加脂肪乳剂1mL/100g体重灌胃,两组均连续灌胃16周后取大鼠的下颌骨下颌支部分检测骨钙、磷及羟脯氨酸含量,取牙槽骨进行脱钙、切片、苏木精-伊红染色,然后用骨形态计量学方法测量牙槽骨松质骨的骨量并计算牙槽骨的骨静态参数。结果:高脂饮食引起大鼠血清中总胆固醇(TC)和低密度脂蛋白胆固醇(LDL-c)明显升高,而高密度脂蛋白胆固醇(HDL-c)则明显降低;脂肪乳剂灌胃使大鼠下颌骨钙?磷及羟脯氨酸含量均下降(P<0.05)。与对照组相比,模型组牙槽骨骨小梁面积百分数(Tb.Ar)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)和骨小梁分离度(Tb.Sp)差异均无统计学意义。结论:长期高脂乳剂灌胃可以造成大鼠下颌骨骨矿物质和骨有机质的丢失,但未能引起大鼠牙槽骨骨量发生明显变化。     

高脂饮食对大鼠下颌骨钙﹑磷及羟脯氨酸含量以及对牙槽骨骨量的影响

目的:探讨高脂饮食与大鼠下颌骨钙、磷及羟脯氨酸含量以及牙槽骨骨量的关系.方法:将20只SD大鼠随机分为正常对照组和脂肪乳剂组,正常对照组予普通饲料及生理盐1 mL/100 g体重灌胃,脂肪乳剂组予普通饮食加脂肪乳剂1 mL/100 g体重灌胃,两组均连续灌胃16周后取大鼠的下颌骨下颌支部分检测骨钙、磷及羟脯氨酸含量,取牙槽骨进行脱钙、切片、苏木精-伊红染色,然后用骨形态计量学方法测量牙槽骨松质骨的骨量并计算牙槽骨的骨静态参数.结果:高脂饮食引起大鼠血清中总胆固醇(TC)和低密度脂蛋白胆固醇(LDL-c)明显升高,而高密度脂蛋白胆固醇(HDL-c)则明显降低;脂肪乳剂灌胃使大鼠下颌骨钙﹑磷及羟脯氨酸含量均下降(P<0.05).与对照组相比,模型组牙槽骨骨小梁面积百分数(Tb.Ar)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)和骨小梁分离度(Tb.Sp)差异均无统计学意义.结论:长期高脂乳剂灌胃可以造成大鼠下颌骨骨矿物质和骨有机质的丢失,但未能引起大鼠牙槽骨骨量发生明显变化.     

矿化液对体外培养的人牙囊细胞碱性磷酸酶活性的影响

目的：探讨矿化液对体外培养的人牙囊细胞中碱性磷酸酶表达的影响。方法：取12～13岁患者因正畸原因拔除的下颌第三磨牙牙囊，原代培养人牙囊细胞，将第5代细胞加入矿化液培养，应用碱性磷酸酶组织化学方法染色、碱性磷酸酶试剂盒检测人牙囊细胞中碱性磷酸酶的活性。结果：培养的牙囊细胞呈长梭形、不规则多角形。免疫细胞化学染色显示抗波形丝蛋白染色阳性。矿化液诱导7d，碱性磷酸酶染色呈阳性，碱性磷酸酶活性检测第3、5、7d明显升高，与对照组均有显著性差异。结论：矿化液能增强体外培养的人牙囊细胞中碱性磷酸酶的表达。     

碱性成纤维细胞生长因子和胰岛素样生长因子-1对人牙髓细胞碱性磷酸酶活性及矿化能力的影响

目的探讨碱性成纤维细胞生长因子(bFGF)和胰岛素样生长因子(IGF)-1对人牙髓细胞碱性磷酸酶活性及矿化能力的影响。方法经10ng/mlbFGF(或)和100ng/mlIGF-1刺激4d后,在410nm波长,检测牙髓细胞的碱性磷酸酶活性值;经矿化液诱导后,采用VonKossa染色法检测不同组细胞的矿化能力。结果培养4d后,IGF-1组及bFGF加IGF-1组均高于对照组(P<0.05),而且bFGF加IGF-1组高于IGF-1组(P<0.05),而bFGF组吸光度值与对照组差异无显著性(P>0.05)。结论IGF-1可促进牙髓细胞向具有矿化能力的细胞分化,而bFGF可能起协同促进作用。     

骨形成蛋白对牙周膜细胞增殖和碱性磷酸酶活性的影响

目的：了解骨形成蛋白（ＢＭＰ）剂量变化对人牙周膜细胞（ＰＤＬＣ）增殖和碱性磷酸酶（ＡＬＰａｓｅ）活性的影响。方法：应用细胞培养技术、噻唑盐比色测定（ＭＴＴ）和酶动力学方法。结果：ＢＭＰ作用后的实验组ＰＤＬＣ的增殖和ＡＬＰａｓｅ活性较对照组均有明显的升高；且有一个合适的剂量范围。ＰＤＬＣ的增殖和ＡＬＰａｓｅ活性受ＢＭＰ浓度梯度影响的变化幅度并不完全一致，表现在所需ＢＭＰ用量不同。结论：可以认为ＢＭＰ能够促进人ＰＤＬＣ的增殖和ＡＬＰａｓｅ活性功能，但各自的影响机制可能还有所差异。     

羟基磷灰石,氢氧化钙对鼠牙髓细胞碱性磷酶活性的影响

检测羟基磷灰石和氢氧化钙颗粒对鼠原代牙髓细胞碱性磷酸酶活性的影响。方法：采用０．５％胰酶－０．１％ＥＤＴＡ消化法获取ＳＤ鼠原代牙髓细胞。细胞接种后，分别加入氢氧化钙和羟基磷灰石颗粒，在７ｄ或１４ｄ分别测定鼠原代牙髓细胞的碱性磷酸酶活性。     

体外培养的成骨细胞中碱性磷酸酶活性的检测方法探讨

检测细胞内碱性磷酸酶（ＡＬＰａｓｅ）活性是体外研究细胞分化及功能的重要手段。实验根据ＡＬＰａｓｅ活性检测的基本原理，探讨了体外培养的成骨细胞中ＡＬＰａｓｅ活性的检测方法。对９６孔细胞培养成骨细胞，以２００μｌ０．０５％ＴｒｉｔｏｎＸ－１００＋超声破碎细胞，以氢氧化钠终止ＡＬＰａｓｅ分解底物的反应。对人胚成骨细胞而言，细胞裂解产物与底物及０．４ｍｏｌ／ＬＮａＯＨ的配比比例以４０∶１００∶１００（μｌ）为宜，有较好的灵敏性和可靠性。     

氢氧化钙对牙髓碱性磷酸酶活性的作用

氢氧化钙可促进牙髓、牙本质修复,但作用机制不清楚。本文观察了氢氧化钙、氯化钙和氢氧化钡对提取的人牙髓组织ALP的体外水解作用和对牙髓细胞ALP活性的作用,结果显示,氢氧化钙可激活ALP的体外水解和牙髓细胞ALP活性,氯化钙对两种反应均表现抑制作用,氢氧化钡激活ALP的体外水解作用,但抑制牙髓细胞ALP活性。     

Serum alkaline phosphatase, calcium, and phosphate levels following clinical use of natural coral

Measurement of alkaline phosphatase enzymatic activity is the most commonly used serum marker to assess bone formation. In this present study serum alkaline phosphatase, calcium, and phosphate were measured in 12 patients where natural coral was implanted in surgical bony defects. Blood samples were obtained preoperatively which served as control, and at 1, 7, 15, and 30 days. No statistically significant increase in serum alkaline phosphatase, calcium, and phosphate was observed.     

羟基磷灰石、氢氧化钙对鼠牙髓细胞碱性磷酸酶活性的影响

目的：检测羟基磷灰石和氢氧化钙颗粒对鼠原代牙髓细胞碱性磷酸酶活性的影响。方法：采用０．５％胰酶－０．１％ＥＤＴＡ消化法获取ＳＤ鼠原代牙髓细胞。细胞接种后，分别加入氢氧化钙和羟基磷灰石颗粒（＜５０μｍ），在７ｄ或１４ｄ分别测定鼠原代牙髓细胞的碱性磷酸酶活性。结果：氢氧化钙抑制牙髓细胞碱性磷酸酶活性与其高浓度抑制细胞生长有关；羟基磷灰石具有良好的生物相容性，但其能够显著抑制牙髓细胞碱性磷酸酶的活性，抑制率（０．６ｍｇ／ｍｌ）在７ｄ时为５４．６％，１４ｄ时为４９．０％。结论：羟基磷灰石颗粒的作用可能与牙髓细胞去分化过程有关，两种材料对牙髓细胞的活性存在着不同的作有机制。     

Cell-bound and extracellular matrix-associated alkaline phosphatase activity in rat periodontal ligament

In previous studies it was noted that alkaline phosphatase (ALP) activity in periodontal ligament does not only seem to be related to cells but may also be associated with the extracellular matrix. In an attempt to clarify this we studied the distribution of the enzyme at the electron microscopic level. In addition, ALPactivity was assessed biochemically following extraction of the ligament with (i) agents dissolving the membrane or splitting the phosphatidylinositol anchor (Triton X-100 or phosphatidylinositol-phospholipase C, respectively), and (ii) a matrix-degrading enzyme cocktail (collagenase, hyaluronidase and elastase). Histochemical observations revealed (a) a heterogeneous distribution of ALP-activity, with highest activity adjacent to the alveolar bone and (b) two pools of activity; one bound to cells and one associated with the collagenous extracelluJar matrix. In line with this were the biochemical data indicating that approximately 10% of the enzyme activity was firmly bound to the extracellular matrix and 90% to plasma membranes. Isoelectric focusing did not reveal differences between the two fractions, both samples yielding a single broad band corresponding with an isoelectric point of about 4.4.     

Effects of ovariectomy on trabecular structures of rat alveolar bone

An association between postmenopausal osteoporosis and tooth loss has been proposed. However, histomorphometrical changes in alveolar bone following estrogen deficiency are rarely reported with data on microtrabecular structural changes. To clarify the relationship between estrogen deficiency and tooth loss, we histomorphometrically analyzed the trabecular structural changes of mandibular alveolar bone in ovariectomized rats. Twenty-four adult female Fischer rats were used. Eight rats were sacrificed on day 0 (baseline). The remaining 16 rats were divided into two groups. One group was ovariectomized bilaterally (OVX) and the other group was subjected to sham surgery (Sham). After administration of tetracycline and calcein, the animals were sacrificed 60 days after surgery. Bone histomorphometry, node-strut analysis and measurement of thickness of alveolar bone proper were performed on the interradicular septum of the first molar on the sagittal surface. The trabecular bone volume and trabecular number of the OVX group were significantly lower than those of the baseline and Sham groups. All of the bone resorptive and formative parameters of the OVX group were significantly higher (about one-and-a-half times) than those of the Sham group. Several osteoclasts were seen lining the irregular, eroded surface facing the bone marrow in the OVX group. Furthermore, the OVX group tended to have low microtrabecular stiffness and showed significantly thinner distal alveolar bone proper than in the baseline and Sham groups. In summary, estrogen deficiency caused osteoporotic changes and thin alveolar bone proper in the interradicular septum of rat first molar. This phenomenon might accelerate destruction of alveolar bone and tooth loss, especially in elderly women affected by periodontal disease.     

牙周优势菌内毒素对人牙周膜细胞增殖和碱性磷酸酶活性的影响

目的：研究牙周优势菌牙龈卟啉菌、中间普氏菌、具核梭杆菌的内毒素对人牙周膜细胞（PDL细胞）增殖和碱性磷酸酶活性（ALP）的影响。方法：采用MTT比色试验及酶动力学方法，测定PDL细胞的增殖和ALP活性。结果：内毒素在10μg/mL、100μg/mL高浓度时，可显著抑制PDL细胞增殖，而在0.01μg/mL、0.1μg/mL低浓度时，则促进PDL细胞增殖；在10μg/mL、100μg/mL可呈浓度依赖性方式抑制PDL细胞ALP活性。结论：内毒素对牙周膜细胞的抑制作用主要和其浓度有关，不同来源内毒素差异并不显著；内毒素可能通过影响PDL细胞功能而影响牙周组织的代谢和修复过程。     

Temporal and local appearance of alkaline phosphatase activity in early stages of guided bone regeneration. A descriptive histochemical study in humans

Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters and it seems to be a prerequisite for normal skeletal mineralization. Also, ALP is the most widely recognized marker of osteoblast phenotypes. By a tissue regenerative technique called Guided Bone Regeneration (GBR), it is possible nowadays to regenerate small bony defects. The aim of the present study was to investigate early events in bone healing and neogenesis by studying histochemically the temporal and local appearance of the marker Alkaline Phosphatase (ALP) in a GBR model system. Nine healthy volunteers (5 males, 4 females, mean age 31.7 years) participated in the experiment. After raising a mucoperiosteal flap from the mandibular second molar to the retromolar area in each volunteer, a hollow titanium test cylinder was placed into a congruent bony bed and the coronal end of the cylinder was closed with an ePTFE-membrane. Then the flap was adapted and sutured to obtain primary wound closure. After 2, 7 and 12 weeks, the regenerated tissue within the cylinders was harvested. Histologically, ALP activity was observed associated with the osteoid seams in the very basal part of the regenerate where new bone trabeculae were in the process of being formed. More coronally, large round cells seemed to secrete an ALP-positive substance since in the center of such cell clusters strong ALP activity located extracellularly was detected. In the present experiment, ALP seemed to have been an early sign of osteoblast secretion of a matrix which subsequently was determined to become osteoid. ALP activity was never seen isolated within connective tissue and away from bone. This is an indication that its source is linked to existing bone. The present study has documented for the first time the appearance of ALP activity in guided bone regenerations in humans. It has revealed that: 1) Osteogenesis in guided bone regeneration is preceded by localized, marked expression of ALP in an organized connective tissue environment. 2) Bone neogenesis is an early event in this experimental setup and may be detected already 2 weeks after wounding. 3) Expression of ALP and subsequent bone neogenesis is originating from and topographically linked to pre-existing bone structures.     

烟草烟雾对大鼠牙槽骨内牙本质基质蛋白1C 末端表达的影响

目的 观察烟草烟雾对大鼠牙槽骨内牙本质基质蛋白（DMP）1 C末端表达的影响。方法 建立试验大鼠吸入烟草烟雾模型,免疫组织化学法检测DMP1 C 末端在大鼠牙槽骨内的表达。结果 阳性对照组：DMP1 C末端深棕黄色连续线形表达于固有牙槽骨的钙化前缘,形成明显的矿化条带;棕黄色颗粒状密布于骨细胞周围,浅棕黄色颗粒状散布于矿化区域;上述表达由牙槽嵴顶至根尖方向逐渐上调。试验组：DMP1 C 末端表达部位同对照组,表达强度下调且随试验时间的延长下调加重。结论 吸入烟草烟雾可下调大鼠牙槽骨内DMP1C 末端的表达;在试验周期内,DMP1 C 末端的表达下调呈时间依赖性。