Comparative study of demineralized collagen degradation determined by hydroxyproline assay and microscopic depth measurement

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• 作者：Md. Sofiqul Islam^a ; ^b ; ^{soheltmd@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; ^{drsohel_bds2000@yahoo.com" class="auth_mail" title="E-mail the corresponding author} ; Sahar Jameel Khunkar^a ; Syozi Nakashima^a ; Alireza Sadr^c ; Toru Nikaido^a ; Junji Tagami^a
• 关键词：Dentin ; Demineralization ; Collagen-degradation ; Hydroxyproline ; Microscopy
• 刊名：Journal of Dentistry
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：47
• 期：Complete
• 页码：94-97
• 全文大小：733 K

文摘

Quantification of collagen degradation is an important parameter to evaluate dentin caries progression or the efficacy of caries prevention aid. The aim of this study was to validate the simple light microscopic technique (LM) to evaluate collagen degradation by comparing with hydroxyproline assay technique (HPN).

Materials and methods

Bovine root dentin blocks were embedded in acrylic resin, polished and covered with nail varnish except a 1.5 × 2.5 mm window. The specimens were demineralized in acetate buffer (pH 4.3) for 3 days to create incipient lesions and were exposed to collagenase enzyme for 6, 9 and 16 h. The specimens were sectioned into thin sections (200–220 μm) to measure the degraded depth of collagen matrix by LM. The enzyme solutions were allocated to HPN assay using the simplified chloramines-T method. Correlation between LM and HPN was evaluated by Pearson correlation analysis. Anti-collagen degradation efficacy of 0.12% chlorhexidine (CHX) was evaluated by LM.

Result

The depths of the degraded collagen and amount of hydroxyproline in 3 exposure periods were 27.8 ± 3.8 μm and 28.7 ± 4.2 μg for 6 h, 48.1 ± 8.6 μm and 45.3 ± 6.1 μg for 9 h, and 74.2 ± 9.7 μm and 71.3 ± 8.0 μg for 16 h, respectively. A significantly positive correlation (r = 0.94, CI: 0.88–0.97, p < 0.0001) was observed between LM and HPN and incubation time showed a linear correlation with amount of collagen degradation (R² = 0.92). The CHX group (28.6 ± 3.3 μm) showed significantly lower collagen degradation than that of control group (53.1 ± 7.8 μm: p < 0.01).

Conclusion

The LM might be a reliable and simplified method to evaluate collagen degradation.