牙本質黏著劑藉由形成混合層，以提供牙本質與樹脂之間的黏著。目前所用的牙科黏著劑¬–酸蝕沖洗或自體酸蝕系統，被認為可以活化金屬基質蛋白酶(MMPs)，破壞混合層的膠原蛋白，而影響黏著。目前已證實黏著力喪失與膠原蛋白的破壞程度有關。而最近的實驗發現自體酸蝕黏著劑還會促使牙本質牙髓系統產生更多的MMP-2。為了去證實MMP-2會於混合層膠原蛋白的影響，本實驗會使用不同黏著系統測試MMP-2在人體內的反應。因此，本實驗目的是希望藉由人體試驗去檢驗兩種黏著系統對MM-2活性的影響與反應。本實驗招募將拔除完整或蛀牙智齒的志願者。除了控制組外，所有牙齒在口內都會修形成一級窩洞，並補上樹脂。在填補前，所有牙齒會經過蒸餾水或 chlohexidine (CHX)前處理一分鐘，並隨機使用酸蝕沖洗(Single bond, 簡稱SB)或自酸蝕系統(SE bond, 簡稱SE)做黏著。一週或一個月後，拔除牙齒、並縱切成片狀，以做zymography、IHC分析使用，分別定性或定量偵測MMP-2。根據實驗變數，所有填補牙齒可分為八組: SB-7, SBc-7, SE-7, SEc-7, SB-30, SBc-30, SE-30, 和 SEc-30。在zymography分析的結果方面，牙髓腔附近活化的MMP-2表現的較黏著區域明顯的多。比較此兩區域組間差異性；黏著區附近MMP-2各組間無明顯差異，但牙髓腔附近的MMP-2表現則具有組間差異。此差異為-- SB黏著的牙齒較SE黏著的牙齒有更多活化的MMP-2，尤其是SB-7這組。而隨時間的影響下:無論有無使用CHX， SB填補的牙齒在30天後，MMP-2表現會逐漸減少；而SE的牙齒則逐漸增加。當使用CHX時，CHX可以有效抑制MMP-2表現，尤其在SB填補的牙齒上。隨著時間抑制的效果: 在SB填補的牙齒上，抑制效果可達30天；而SE填補牙齒上，當至30天時則不見其影響力。 在免疫組織染色的部分，得到與zymography類似的結果。牙髓腔附近牙本質有較多活化的MMP-2，且MMP-2集中於牙本前質與牙本前質附近的牙本質基質。牙本前質的部分—SB-7最明顯，其次是SE-7、SB-30、SBc-30，其餘沒有明顯差異；牙本質基質部分—SB-7最明顯，其次是SBc-7，其餘沒顯著差異。使用CHX、在組織學表現上，只有在7天的組別可以看到抑制效果。
不同牙本質黏著系統以不同模式影響牙本質內MMP-2表現；30天內，SB的影響又較SE大。而CHX在7天時，可以抑制MMP-2形成，但30天後，只對SB有效。相較於SE，SB抑制效果較大。

Adhesive systems provide resin-dentin adhesion by formation of a hybrid layer. Etch-and-rinse and self-etch systems, have been reported to activate matrix metalloproteinases (MMPs), which may degrade collagen in the hybrid layer and compromise the adhesive quality. The correlation between the collagen degradation and loss of bond strength has been ensured. Recently, more MMP-2 expression in dentin pulp complex responding to self-etch systems has been reported. To confirm theoretical collagen degradation by MMP-2, the effect of different adhesive systems on MMP activation should be evaluated under functional condition. The objective of this in vivo study was to examine the degree of MMP-2 activation by different adhesive systems. With study protocol approved by the ethical committee, volunteers with non-carious third molars were enrolled in this study. Class I cavities were prepared, treated with one of different combined protocols (Single bond vs. SE bond, pretreatment with distilled water vs. chlorhexidine), and then restored with resin composite. After 7-day or 30-day oral function, teeth were extracted and sectioned into several slices. For zymography, coronal dentin from sliced teeth was divided into bond and pulp areas. Active MMP-2 of both areas was examined individually. The other slices were prepared for immunohistochemistry (IHC) to reveal the localization of MMP-2. Based on adhesive systems, use of chlorhexidine, and retained days, restored teeth were assigned to groups SB-7, SBc-7, SE-7, SEc-7, SB-30, SBc-30, SE-30, and SEc-30.From results of zymography, active MMP-2 of pulp area was more relevant than that of bond area. No significant difference was found among bond areas of all groups. On pulp areas, SB bonded teeth had more MMP-2 expression than that of SE bonded teeth, especially at Day 7. Abundant MMP-2 of SB treated teeth was detected at Day 7 and decreased at Day 30 while MMP-2 in SE bonded teeth rose gradually with time. Chlorhexidine-treated groups also exhibited less active MMP-2 at Day 7 regardless of adhesive systems. The effect of chlorhexidine was found up to 30 day in SBc-30. In the result of IHC, intense MMP-2 immunoreactivity was localized in predentin and dentinal matrix adjacent to pulp area. SB-7 showed the most immunoreactivity while SE-7, SBc-7, SB-30, and SBc-30 showed intermediate immunoreactivity. The others revealed no significant difference. Two adhesive systems affected in vivo active MMP-2 expression of pulp area in different degrees. Chlorhexidine appeared to inhibit active MMP-2 expression of pulp areas in group SBc-7, SEc-7, and SEc-30. Extended effect of CHX in SBc-30 may arise from CHX retention. The IHC finding almost responds to result of zymogarphy and demonstrate that most MMP-2 detected in pulp area may come from predentin.

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