Phytomedicine 20 (2013) 1203–1210 Contents lists available at ScienceDirect Phytomedicine j ourna l h o mepage: www.elsevier.de/phymed Berberine’s effect on periodontal tissue degradation by matrix metalloproteinases: an in vitro and in vivo experiment Hsiao-Pei Tu a,b , Martin M.J. Fu c , Po-Jan Kuo a , Yu-Tang Chin a , Cheng-Yang Chiang a , Cheng-Long Chung a , Earl Fu a,∗ a Department of Periodontology, School of Dentistry, National Defense Medical Center and Tri-Service General Hospital, Taipei, Taiwan, ROC b Department of Dental Hygiene, China Medical University, Taichung, Taiwan, ROC c Resident, Division of Periodontology, Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, USA a r t i c l e i n f o Keywords: berberine periodontitis matrix metalloproteinase fibroblast macrophage a b s t r a c t Periodontal disease involves tissue destruction caused by interactions among bacterial antigens and inflammatory mediators including matrix metalloproteinases (MMPs). Berberine, an isoquinoline alka- loid isolated from medicinal herbs, can inhibit the degradative action of extracellular MMPs. The effect of berberine on the periodontal expression of MMPs was examined in vitro and in vivo. Gelatinolytic activity of pro-MMP-2, MMP-2, and MMP-9 in the human gingival fibroblast and/or U-937 was compared after treatment with Porphyromonas gingivalis lipopolysaccharide (P.g. LPS) in four medias containing 0, 1, 10 and 100 M of berberine each. Twelve animals were divided into three groups for the study: (A) non- ligation, (B) ligation, and (C) ligation-plus-berberine (75 mg/kg berberine by gastric lavage daily); and the effect of berberine on periodontal destruction was evaluated in the ligature-induced periodontitis in rats for 8 days by micro computerized tomography (micro-CT), histology and immunohistochem- istry (IHC). An enhancing effect of P.g. LPS on MMP activities was identified, with a greater effect on fibroblasts/U937 co-culture than on either culture alone. When berberine was added to the LPS-treated cultures, the activities of MMPs were significantly reduced in dose-dependent manner. In the animals, the trends of the following parameters were compared. 1. Micro-CT distances between cemento-enamel junction (CEJ) and dental alveolar bone crest: B > C > A. 2. Histometrically measured crest bone levels: B > C > A. 3. Amount of collagen deposited in tissue areas: A > C > B. 4. Attachment loss: B > C ≈ A. 5. Con- nective tissue (CT) attachment: B > either A or C. 6. Expression of cells stained positive for MMP-2 and -9 by IHC: B > C > A. In conclusion, berberine demonstrated in vitro an inhibitory effect on P.g. LPS-enhanced MMP activities of HGF and U937 macrophages, reducing in vivo gingival tissue degradation in periodon- titic rats. We thus propose that berberine may slow periodontal degradation through the regulation of MMPs in periodontitis induced by bacterial plaque. © 2013 Elsevier GmbH. All rights reserved. Introduction Periodontal disease is an inflammatory disorder in which inflammatory reaction may damage surrounding cells and tis- sue structures including the alveolar bone, causing tooth loss. In the inflamed periodontal tissues, dental plaque bacteria and their Abbreviations: MMP, Matrix metalloproteinase; P.g., porphyromonas gingivalis; LPS, lipopolysaccharide; micro-CT, Micro Computed Tomography; CEJ, cemento- enamel junction; IHC, immunohistochemistry; CT, connective tissue; HGF, human gingival fibroblasts. ∗ Corresponding author at: Department of Periodontology, School of Dentistry, National Defense Medical Center, PO Box 90048-507, Taipei, Taiwan, ROC. Tel.: +886 2 87927150; fax: +886 2 87927145. E-mail address: dentalab@tpts5.seed.net.tw (E. Fu). products induce polymorphonuclear leukocyte infiltration, edema and vascular dilatation (Page, 1991). Complex interactions among various inflammatory mediators and tissue degradation have been carefully examined in vitro and in vivo and suggested to be involved in the pathogenic mechanisms of periodontitis (Kuo et al., 2012; Sundararaj et al., 2009; Fiehn et al., 1992; Di Paola et al., 2004). Berberine, an isoquinoline alkaloid, can be isolated from many medicinal herbs, such as Rhizoma coptidis (Huanglian), Hydrastis canadensis (goldenseal) and Cortex phellodendri (Huangbai) (Ikram, 1975). Berberine-containing plants are used medicinally in many traditional medical systems, including Chinese herbal and Ayurvedic herbal medicine (Imanshahidi and Hosseinzadeh, 2008). Studies have indicated that berberine has multiple pharmacolog- ical activities including anti-inflammatory, anti-cyclooxygenase, and anti-inducible nitric oxide synthases effects (Jiang et al., 2011; Lee et al., 2007). Recently, the effect of berberine against 0944-7113/$ – see front matter © 2013 Elsevier GmbH. All rights reserved. http://dx.doi.org/10.1016/j.phymed.2013.06.001