正畸牙周组织改建中细胞外基质代谢调节机制的研究
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摘要
正畸牙齿移动过程中，牙周组织的改建是牙周组织细胞对机械力产生
生物学反应的结果。具体表现为牙周组织细胞外基质（extracellur matrix，
ECM）的降解与合成，通过ECM降解与合成的动态平衡实现牙齿矫正
的目的。但到目前为止，有关机械力作用下，牙周组织ECM代谢的分子
基础及调节机制人们还并不清楚。基质金属蛋白酶/金属蛋白酶组织抑制
因子（matrix metalloproteinases/tissue inhibitor of metalloproteinases，
MMPs/TIMPs）系统对ECM的代谢进行直接的调控，而这种调控作用又
受局部细胞因子的影响。MMP-3属MMPs家族中基质溶解素类的一种，
又称基质溶解素—1（stromelysin-1），其作用底物广泛，不仅可以单独降
解构成ECM的大部分成份，同时可以激活MMP-1、MMP-8及MMP-9
等基质金属蛋白酶的活性，共同参与ECM的代谢过程。因此，围绕着
MMP-3的表达与调控，本研究通过动物实验和细胞培养，结合细胞力学
实验方法和分子生物学测试技术，首次观察了在机械力及白细胞介素—6
作用下，牙周膜成纤维细胞和破骨细胞MMP-3表达变化的情况，以进一
步阐明正畸牙周组织改建过程中细胞外基质代谢调节的分子机制，为揭
示正畸矫治机理，探索临床干预的可能性和方法提供一条新思路。
本研究主要作了以下几方面的工作：
一：MMP-3及TIMP-1在大鼠正畸牙周组织改建过程中的表达
目的：观察大鼠正畸牙周组织改建过程中基质金属蛋白酶—3
（MMP-3）和金属蛋白酶组织抑制因子-1（TIMP-1）表达的变化，探讨
MMP-3及TIMP-1与正畸牙齿移动的关系。方法：在SD成年大鼠上颌
第口旱巴大学馋士学优饵文
右侧第一磨牙与上颌切牙之间安置正畸装置，建立大鼠磨牙移动实验模
型。于牙齿移动1、3、5、7、14天后取材分别进行免疫组化染色、图像
分析。结果：抛移动1天后，牙周组织细胞MMP3表达增强，5天后
MMP－3表达达到高峰，此时破骨细胞胞浆亦呈强阳性表达。以后MMP．3
表达有所下降，但仍高与对照组。而TIMP－l于牙齿移动3天后表达开始
增强，7天后显著表达。结论：MMP习及11MP－l参与了正畸牙周组织
改建过程，MMPS在破骨细胞性骨吸收中起着重要作用。
二：周期性牵张力对人牙周膜细胞MMI，3及TIMP4 的影响
目的：探服期性牵张力又屯厂牙周膜细胞 M3及 11：MP蛋白表
达的影响，进一步阐明正畸牙周组织改酣程中ECM代猢节的肝机
制，e：通过体外细胞培养加载系统，选择频率为 6周／b，弹性基底
膜发生12％形变率的周期性牵张力，利用免疫组化及夹心EtoA检测技
术，观察HPDLC表达MMP－3及TIMP－l的相对强度。结果：HPDLC在
体外表达MMP－3及TIMP－l。加载3天后，MMP－3表达明显增强，与对
照组相比具有显著性差异。而TIMP－l对机械力无应答。结论：在一定条
件下，周期性牵张力可显著影响HPDLC MMP3蛋白的表达。为机枷
．作用下W3参与牙周组织ECM代谢提供了直接证据。
三：白娜介盆一6…膜细胞M3 InRNA及蛋白秘的影响
目的：观察 IL—6对人牙周膜细胞W3蛋白及 W表达的影
响，探讨IL—6介导 M对牙周组织 ECM代狮响的机制。施：
在体外培养耕下，鹏不同浓度IL—6作用腿是否对人牙周膜细胞
MMP3表达的情况产生影响。采用原位杂交、赃组化及蛋白印迹分析
技术对HPDLC表达WP习的情况进行检测。结果：当IL—6的浓度为
邑
10w加及11＊M时，可显著删厨周膜细胞MM卜31lllUvA及蛋
白的表达，且熊明显的浓度喇舰。结论：IL—6的刺激作用，可使
HPDLC表达 MMP3阳性信号增强，表明 IL一6介导 MMP－3参与调节牙
周膜细胞外基质代谢。
四：IL一6体外诱导成年大鼠骨瞩破骨细胞形成的实验观奈
．3．
第口罩巨大钞博士学让怕工
目的：探讨几一6对成年大鼠骨髓破骨细胞生成诱导的影响。施：
采用12周龄SD大鼠无菌条件下取出股骨，剪去两肪端，以a—MEM
将骨髓细胞冲出，然后将细胞悬液接种于预置盖玻片或牙本质片的24 IL
培赧内，次日实验鞭含 IL—6 （ OUhlil）的 Q—MEM培养液继续培
养2周，对照组培养液不含IL—6。结果：培养一周左右光镜下观察实验
组可见明显单核细胞融合成多核细胞现象，多核细胞数量增多。经抗酒
石酸酸性磷酸酶（TRAP）染色呈阳性反应。电镜下可观察到典型的破骨
细胞骨吸收现象。结论 几一6门）可明显诱导成年大鼠?
Periodontal tissue remodeling during orthodontic tooth movement result
from the cellular biological responses of connective tissue to exogenous
mechanical forces. Among these responses, the degradation and synthesis of
extracellular matrix (ECM) take place. But, the identification of the molecular
basis as well as the mechanisms on the modification of ECM in these
processes are poorly understood. Previous studies indicate that matrix
metalloproteinases/tissue inhibitor of metalloproteinases (MMPs/TIMPs)
system played a key role in ECM metabolism. Dynamic equilibrium of MMPs
and TIMPs makes it possible to keep normal tissue remodelling
physiologically, that is at least in part regulated by local cytokines. MIMP-3
(Stromelysin-l), one prominent member of the family of MMPs, can not
only degrade solely the most components of ECM, but also can activate
MMP-l, MMP-8 and MMP-9. The regulation of their activation by MMP-3
may be important in the overall regulation of the connective tissue degradation
in both physiologic and pathologic conditions. Thus, this study centers on
the effects of mechanical forces and interleukin-6 (IL-6) on the expression of
MMP-3 by periodontal ligament cells and osteoclasts in vivo and in vitro, to
elucidate the mechanisms on the metabolism of ECM associated with tooth
movement and enhance our understanding of the biological mechanisms
involved in tissue remodelling that occurs during orthodontic treatment.
.5.
This study mainly contains the following works:
1: The expression of MMP-3 and TIMP-1 during orthodontic tooth
movement in rats.
Objective: To observe the change of matrix metalloproteinase-3
(MMP-3) and tissue inhibitor of metalloproteinase- 1 (TIMP- 1) expression
during experimental tooth movement in rats ,learn about the relationship
between MMP-3, TIMP-l and orthodontic periodontal tissue remodeling.
Methods: Orthodontic appliance was placed between the maxillary right first
molar and maxillary central incisors of adult SD rat. Established rat molar
movement model. Immunohistochemistry and image analyses were performed
at 1-. 3?5~. 7~. 14 days after orthodontic force application Results: The
expression of MMP-3 and TIMP-l were weak immunopostive in normal
periodotium. At ld after tooth movement, the immunoreactivities of MMP-3
were increased, at 5d , reached the maximum. Osteoclasts were also observed
strong positive. The expression of TIIvIP- 1 was increased at 3 days. At 7 days,
a marked increase in TIMP- I -positive firblasts and osteoblasts was found not
only on the pressue side but also on the tension side of the periodontium.
Conclusion: MMP-3 and TIMP-l participate in orthodontic periodontal tissue
remodeling during tooth movement MMP-3 may play an important role in
osteoclastic bone resorption.
2:Effects of cyclic-tension force on the expression of MMP-3 and TIMP-1
by human periodontal ligament celis.
Objective: To investigate the effects of the mechanical stretching on the
expression of MMP-3 and TIMP- I in human periodontal ligament cells
(HPDLC), and to unveil the molecular mechanism of ECM metabolism during
orthodontic periodontal tissue remodeling. Methods: HPDLC were subjected
to 12% elongation by strain unit at 6cycles/min (i.e. 5-s elongation and 5-s
relaxation) for 1, 3and 5days in this experiment. Expression and secretion of
.6.
生物学反应的结果。具体表现为牙周组织细胞外基质（extracellur matrix，
ECM）的降解与合成，通过ECM降解与合成的动态平衡实现牙齿矫正
的目的。但到目前为止，有关机械力作用下，牙周组织ECM代谢的分子
基础及调节机制人们还并不清楚。基质金属蛋白酶/金属蛋白酶组织抑制
因子（matrix metalloproteinases/tissue inhibitor of metalloproteinases，
MMPs/TIMPs）系统对ECM的代谢进行直接的调控，而这种调控作用又
受局部细胞因子的影响。MMP-3属MMPs家族中基质溶解素类的一种，
又称基质溶解素—1（stromelysin-1），其作用底物广泛，不仅可以单独降
解构成ECM的大部分成份，同时可以激活MMP-1、MMP-8及MMP-9
等基质金属蛋白酶的活性，共同参与ECM的代谢过程。因此，围绕着
MMP-3的表达与调控，本研究通过动物实验和细胞培养，结合细胞力学
实验方法和分子生物学测试技术，首次观察了在机械力及白细胞介素—6
作用下，牙周膜成纤维细胞和破骨细胞MMP-3表达变化的情况，以进一
步阐明正畸牙周组织改建过程中细胞外基质代谢调节的分子机制，为揭
示正畸矫治机理，探索临床干预的可能性和方法提供一条新思路。
本研究主要作了以下几方面的工作：
一：MMP-3及TIMP-1在大鼠正畸牙周组织改建过程中的表达
目的：观察大鼠正畸牙周组织改建过程中基质金属蛋白酶—3
（MMP-3）和金属蛋白酶组织抑制因子-1（TIMP-1）表达的变化，探讨
MMP-3及TIMP-1与正畸牙齿移动的关系。方法：在SD成年大鼠上颌
第口旱巴大学馋士学优饵文
右侧第一磨牙与上颌切牙之间安置正畸装置，建立大鼠磨牙移动实验模
型。于牙齿移动1、3、5、7、14天后取材分别进行免疫组化染色、图像
分析。结果：抛移动1天后，牙周组织细胞MMP3表达增强，5天后
MMP－3表达达到高峰，此时破骨细胞胞浆亦呈强阳性表达。以后MMP．3
表达有所下降，但仍高与对照组。而TIMP－l于牙齿移动3天后表达开始
增强，7天后显著表达。结论：MMP习及11MP－l参与了正畸牙周组织
改建过程，MMPS在破骨细胞性骨吸收中起着重要作用。
二：周期性牵张力对人牙周膜细胞MMI，3及TIMP4 的影响
目的：探服期性牵张力又屯厂牙周膜细胞 M3及 11：MP蛋白表
达的影响，进一步阐明正畸牙周组织改酣程中ECM代猢节的肝机
制，e：通过体外细胞培养加载系统，选择频率为 6周／b，弹性基底
膜发生12％形变率的周期性牵张力，利用免疫组化及夹心EtoA检测技
术，观察HPDLC表达MMP－3及TIMP－l的相对强度。结果：HPDLC在
体外表达MMP－3及TIMP－l。加载3天后，MMP－3表达明显增强，与对
照组相比具有显著性差异。而TIMP－l对机械力无应答。结论：在一定条
件下，周期性牵张力可显著影响HPDLC MMP3蛋白的表达。为机枷
．作用下W3参与牙周组织ECM代谢提供了直接证据。
三：白娜介盆一6…膜细胞M3 InRNA及蛋白秘的影响
目的：观察 IL—6对人牙周膜细胞W3蛋白及 W表达的影
响，探讨IL—6介导 M对牙周组织 ECM代狮响的机制。施：
在体外培养耕下，鹏不同浓度IL—6作用腿是否对人牙周膜细胞
MMP3表达的情况产生影响。采用原位杂交、赃组化及蛋白印迹分析
技术对HPDLC表达WP习的情况进行检测。结果：当IL—6的浓度为
邑
10w加及11＊M时，可显著删厨周膜细胞MM卜31lllUvA及蛋
白的表达，且熊明显的浓度喇舰。结论：IL—6的刺激作用，可使
HPDLC表达 MMP3阳性信号增强，表明 IL一6介导 MMP－3参与调节牙
周膜细胞外基质代谢。
四：IL一6体外诱导成年大鼠骨瞩破骨细胞形成的实验观奈
．3．
第口罩巨大钞博士学让怕工
目的：探讨几一6对成年大鼠骨髓破骨细胞生成诱导的影响。施：
采用12周龄SD大鼠无菌条件下取出股骨，剪去两肪端，以a—MEM
将骨髓细胞冲出，然后将细胞悬液接种于预置盖玻片或牙本质片的24 IL
培赧内，次日实验鞭含 IL—6 （ OUhlil）的 Q—MEM培养液继续培
养2周，对照组培养液不含IL—6。结果：培养一周左右光镜下观察实验
组可见明显单核细胞融合成多核细胞现象，多核细胞数量增多。经抗酒
石酸酸性磷酸酶（TRAP）染色呈阳性反应。电镜下可观察到典型的破骨
细胞骨吸收现象。结论 几一6门）可明显诱导成年大鼠?
Periodontal tissue remodeling during orthodontic tooth movement result
from the cellular biological responses of connective tissue to exogenous
mechanical forces. Among these responses, the degradation and synthesis of
extracellular matrix (ECM) take place. But, the identification of the molecular
basis as well as the mechanisms on the modification of ECM in these
processes are poorly understood. Previous studies indicate that matrix
metalloproteinases/tissue inhibitor of metalloproteinases (MMPs/TIMPs)
system played a key role in ECM metabolism. Dynamic equilibrium of MMPs
and TIMPs makes it possible to keep normal tissue remodelling
physiologically, that is at least in part regulated by local cytokines. MIMP-3
(Stromelysin-l), one prominent member of the family of MMPs, can not
only degrade solely the most components of ECM, but also can activate
MMP-l, MMP-8 and MMP-9. The regulation of their activation by MMP-3
may be important in the overall regulation of the connective tissue degradation
in both physiologic and pathologic conditions. Thus, this study centers on
the effects of mechanical forces and interleukin-6 (IL-6) on the expression of
MMP-3 by periodontal ligament cells and osteoclasts in vivo and in vitro, to
elucidate the mechanisms on the metabolism of ECM associated with tooth
movement and enhance our understanding of the biological mechanisms
involved in tissue remodelling that occurs during orthodontic treatment.
.5.
This study mainly contains the following works:
1: The expression of MMP-3 and TIMP-1 during orthodontic tooth
movement in rats.
Objective: To observe the change of matrix metalloproteinase-3
(MMP-3) and tissue inhibitor of metalloproteinase- 1 (TIMP- 1) expression
during experimental tooth movement in rats ,learn about the relationship
between MMP-3, TIMP-l and orthodontic periodontal tissue remodeling.
Methods: Orthodontic appliance was placed between the maxillary right first
molar and maxillary central incisors of adult SD rat. Established rat molar
movement model. Immunohistochemistry and image analyses were performed
at 1-. 3?5~. 7~. 14 days after orthodontic force application Results: The
expression of MMP-3 and TIMP-l were weak immunopostive in normal
periodotium. At ld after tooth movement, the immunoreactivities of MMP-3
were increased, at 5d , reached the maximum. Osteoclasts were also observed
strong positive. The expression of TIIvIP- 1 was increased at 3 days. At 7 days,
a marked increase in TIMP- I -positive firblasts and osteoblasts was found not
only on the pressue side but also on the tension side of the periodontium.
Conclusion: MMP-3 and TIMP-l participate in orthodontic periodontal tissue
remodeling during tooth movement MMP-3 may play an important role in
osteoclastic bone resorption.
2:Effects of cyclic-tension force on the expression of MMP-3 and TIMP-1
by human periodontal ligament celis.
Objective: To investigate the effects of the mechanical stretching on the
expression of MMP-3 and TIMP- I in human periodontal ligament cells
(HPDLC), and to unveil the molecular mechanism of ECM metabolism during
orthodontic periodontal tissue remodeling. Methods: HPDLC were subjected
to 12% elongation by strain unit at 6cycles/min (i.e. 5-s elongation and 5-s
relaxation) for 1, 3and 5days in this experiment. Expression and secretion of
.6.
引文
1． Somerman MJ, Young MF, Foster RA, etal. Characteristics of human periodontal ligament cells in vitro. Arch Oral Biol,1990;35:241-247
2． Carnes DL, Maeder CL, Graves DT. Cells with Osteoblastic phenotypes can be explanted from human gingiva and periodontal ligament. J Periodontal, 1997;68:70-707
3． Somerman MJ, Archer SY, Imm GR, etal. A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res 1998;67:66-70
4． Mcculloch CAG, Lekic P, Mckee MD. Role of physical forces in regulating the form and function of the periodontal ligament. Periodontology 2000, 2000;24;56-72
5． Karimbux NY, Ramamurthy NS, Golub LM, etal. The expression of collagen and mRNA in porphyromonas gingivalis-induced periodontitis in rats: The effect of doxycycline and chemically modified tetracycline.
J periodontal, 1998,69(1):34-40
6、方石岗、杨继震。基质金属蛋白酶与组织创伤修复。中华创伤杂志，1999：15(5)：395-396
7、高庆、吴秉铨。基质金属蛋白酶与肿瘤侵袭和转移。中华病理学杂志，1998：27(2)：155-156
8、Stetler-Stevenson WG, Hewitt R, Corcoran M. Matrix metalloprateinases and tumor invasion:from correlation and causality to clinic. Scm in Cancer Bio, 1996;7:147-154
9、李成章、任铁冠。基质金属蛋白酶家族。口腔医学纵横杂志，1998；14(1)：50-52。
10、Bolcato-Bellemin AL, Elkaim R, Abehsera A, etal. Expression of mRNAs encoding for α and β integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts. J Dent Res 2000;79(9):1712-1716
11、王宇、黄宇琦。Ito细胞及金属蛋白酶与肝内基质降解。医学综述，1999；5(2)：78-80
12、邱明、张伟。金属蛋白酶及其抑制因子与肿瘤袭转移。医学综述，1999：5(7)：293-294
13、Yamaguchi M, Shimizu N, Ozawa Y, etal. Effect of tension-force on plasminogen activator activity from human periodontal ligament cells, J Periodont Res1997;32:308-314
14、Ogata Y, Enghild JJ, Nagase H. Matrix metalloproteinase3 (stromelysin) activates the precursors for human matrix metalloproteinase 9. J Bio Chem 1992;267:3581-3584
15、Mohd H, Said S, Sander L, etal. GCF levels of MMP-3 and MMP-8 following placement of bioresorbable membranes, J Clin Periodontol 1999;26:757-763
16、高庆、吴秉铨、郑杰等。Ⅰ型膜型基质金属蛋白酶在癌细胞系中的表达及其与明胶酶A活化关系的研究。中华病理学杂志，1999；28(3)：191-193
17、Lukinmaa PL, Vaahtokari A,Vainio S,etal. Expression of type Ⅰ collagen pro- α_2 chain Mrna in adult human permanent teeth as revealed by in situ hybridization. J Dent Res 1992;71(1):36-42
18、Ryan EM,Golub LM.Modulation of matrix metalloproteinase activities in periodontitis as a treatment strategy. Periodontology 2000, 2000;24:226-236
19、Meikle MC, Hembry RM, Holley J, etal. Immunolocalization of matrix metalloproteinases and TIMP-1 (tissue inhibitor of metalloproteinases) in human gingival tissues from periodontitis patients. J Periodont Res 1994;29:118-126
20、Ryan M,Ramamurthy N,Golub L. Matrix metalloproteinases and their inhibition in periodontal treatment. Curr Opin Periodontol, 1996;3:85-96
21、付云。金属蛋白酶组织抑制因子对牙周病的抑制作用。国外医学口腔医学分册，1999；26(1)：33-35
22、Richards D, Rutherford RB, Intefieukin-1 regulation of procollagenase mRNA and protein in periodontal fibroblasts in vitro. J Periodont Res 1996;25:222-229
23、Nakaya H, Oates TW, Hoang AM, etal .Effect of interleukin-1 on matrix metalloproteinase-3 levels in human periodontal ligament cells. J Periodontol 1997;68:517-523
24、Nakaya H,Osawa G,Iwasaki N,etal. Effect of bisphonate on matrix metalloproteinase enzymes in human periodontal ligament cells.J Periodontol,2000;71:1158-1161
25、Golub L, Sorsa T, Lee HM, etal. Doxycyclinein hibits neutrophil(PMN)-type matrix metalloproteinases in human adult periodontitis gingiva. J Clin Periodontol, 1995;21:1-9
26、Yamaguchi M, Shimizu N, Shibata Y, etal. Effects of different magnitudes of tension-force on alkaline phosphatase activity in periodontal ligament cells. J Dent Res 1996;75(3):889-894
27、Nakago-Matsuo C, Matsuo T, Nakago T. Intracellular calcium response to hy draulic pressure in human periodontal ligament fibroblasts. Am J Orthod Dentofac Orthop 1996;109:244-248
28、Yamaguchi M, Shimizu N, Goseki T, etal. Effect of different magnitudes of tension force on prostaglandin E2 production by human periodontal ligament cells. Archs Oral Biol 1994;39:877-884
29、Shimizu N, Yamaguchi M, Goseld T, etal. Cyclic-tension force stimulates interleukin-1 production by human periodontal ligament cells. J Periodont Res 1994;29:328-333
30、任蕾、程祥荣、沈真祥。牙周膜胶原蛋白的更新与牙移动。国外医学口腔医学分册 2000；27(3):168-171
31、Howard PS, Kucich U, Taliwal R, etal. Mechanical forces alter extracellular matrix synthesis by human periodontal ligament fibroblasts. J Periodontol Res, 1998;33:500-508
2． Carnes DL, Maeder CL, Graves DT. Cells with Osteoblastic phenotypes can be explanted from human gingiva and periodontal ligament. J Periodontal, 1997;68:70-707
3． Somerman MJ, Archer SY, Imm GR, etal. A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res 1998;67:66-70
4． Mcculloch CAG, Lekic P, Mckee MD. Role of physical forces in regulating the form and function of the periodontal ligament. Periodontology 2000, 2000;24;56-72
5． Karimbux NY, Ramamurthy NS, Golub LM, etal. The expression of collagen and mRNA in porphyromonas gingivalis-induced periodontitis in rats: The effect of doxycycline and chemically modified tetracycline.
J periodontal, 1998,69(1):34-40
6、方石岗、杨继震。基质金属蛋白酶与组织创伤修复。中华创伤杂志，1999：15(5)：395-396
7、高庆、吴秉铨。基质金属蛋白酶与肿瘤侵袭和转移。中华病理学杂志，1998：27(2)：155-156
8、Stetler-Stevenson WG, Hewitt R, Corcoran M. Matrix metalloprateinases and tumor invasion:from correlation and causality to clinic. Scm in Cancer Bio, 1996;7:147-154
9、李成章、任铁冠。基质金属蛋白酶家族。口腔医学纵横杂志，1998；14(1)：50-52。
10、Bolcato-Bellemin AL, Elkaim R, Abehsera A, etal. Expression of mRNAs encoding for α and β integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts. J Dent Res 2000;79(9):1712-1716
11、王宇、黄宇琦。Ito细胞及金属蛋白酶与肝内基质降解。医学综述，1999；5(2)：78-80
12、邱明、张伟。金属蛋白酶及其抑制因子与肿瘤袭转移。医学综述，1999：5(7)：293-294
13、Yamaguchi M, Shimizu N, Ozawa Y, etal. Effect of tension-force on plasminogen activator activity from human periodontal ligament cells, J Periodont Res1997;32:308-314
14、Ogata Y, Enghild JJ, Nagase H. Matrix metalloproteinase3 (stromelysin) activates the precursors for human matrix metalloproteinase 9. J Bio Chem 1992;267:3581-3584
15、Mohd H, Said S, Sander L, etal. GCF levels of MMP-3 and MMP-8 following placement of bioresorbable membranes, J Clin Periodontol 1999;26:757-763
16、高庆、吴秉铨、郑杰等。Ⅰ型膜型基质金属蛋白酶在癌细胞系中的表达及其与明胶酶A活化关系的研究。中华病理学杂志，1999；28(3)：191-193
17、Lukinmaa PL, Vaahtokari A,Vainio S,etal. Expression of type Ⅰ collagen pro- α_2 chain Mrna in adult human permanent teeth as revealed by in situ hybridization. J Dent Res 1992;71(1):36-42
18、Ryan EM,Golub LM.Modulation of matrix metalloproteinase activities in periodontitis as a treatment strategy. Periodontology 2000, 2000;24:226-236
19、Meikle MC, Hembry RM, Holley J, etal. Immunolocalization of matrix metalloproteinases and TIMP-1 (tissue inhibitor of metalloproteinases) in human gingival tissues from periodontitis patients. J Periodont Res 1994;29:118-126
20、Ryan M,Ramamurthy N,Golub L. Matrix metalloproteinases and their inhibition in periodontal treatment. Curr Opin Periodontol, 1996;3:85-96
21、付云。金属蛋白酶组织抑制因子对牙周病的抑制作用。国外医学口腔医学分册，1999；26(1)：33-35
22、Richards D, Rutherford RB, Intefieukin-1 regulation of procollagenase mRNA and protein in periodontal fibroblasts in vitro. J Periodont Res 1996;25:222-229
23、Nakaya H, Oates TW, Hoang AM, etal .Effect of interleukin-1 on matrix metalloproteinase-3 levels in human periodontal ligament cells. J Periodontol 1997;68:517-523
24、Nakaya H,Osawa G,Iwasaki N,etal. Effect of bisphonate on matrix metalloproteinase enzymes in human periodontal ligament cells.J Periodontol,2000;71:1158-1161
25、Golub L, Sorsa T, Lee HM, etal. Doxycyclinein hibits neutrophil(PMN)-type matrix metalloproteinases in human adult periodontitis gingiva. J Clin Periodontol, 1995;21:1-9
26、Yamaguchi M, Shimizu N, Shibata Y, etal. Effects of different magnitudes of tension-force on alkaline phosphatase activity in periodontal ligament cells. J Dent Res 1996;75(3):889-894
27、Nakago-Matsuo C, Matsuo T, Nakago T. Intracellular calcium response to hy draulic pressure in human periodontal ligament fibroblasts. Am J Orthod Dentofac Orthop 1996;109:244-248
28、Yamaguchi M, Shimizu N, Goseki T, etal. Effect of different magnitudes of tension force on prostaglandin E2 production by human periodontal ligament cells. Archs Oral Biol 1994;39:877-884
29、Shimizu N, Yamaguchi M, Goseld T, etal. Cyclic-tension force stimulates interleukin-1 production by human periodontal ligament cells. J Periodont Res 1994;29:328-333
30、任蕾、程祥荣、沈真祥。牙周膜胶原蛋白的更新与牙移动。国外医学口腔医学分册 2000；27(3):168-171
31、Howard PS, Kucich U, Taliwal R, etal. Mechanical forces alter extracellular matrix synthesis by human periodontal ligament fibroblasts. J Periodontol Res, 1998;33:500-508