In vitro-activity of oily calcium hydroxide suspension on microorganisms as well as on human alveolar osteoblasts and periodontal ligament fibroblasts

BMC Oral Health

Anton Sculean¹  Richard J Miron¹  Tatjana Strugar¹  Sigrun Eick¹ 

[1] Department of Periodontology, School of Dental Medicine, University of Bern, CH-3010, Bern, Switzerland

关键词: Periodontopathogens;    Periodontal ligament fibroblasts;    Human alveolar osteoblasts;    Oily calcium hydroxide suspension;   

Others  :  1121475
DOI  :  10.1186/1472-6831-14-9

 received in 2013-10-14, accepted in 2014-01-20,  发布年份 2014

【 摘 要 】

Background

Findings from animal and human studies have indicated that an oily calcium hydroxide suspension (OCHS) may improve early wound healing in the treatment of periodontitis. Calcium hydroxide as the main component is well known for its antimicrobial activity, however at present the effect of OCHS on the influence of periodontal wound healing/regeneration is still very limited. The purpose of this in vitro study was to investigate the effect of OCHS on periodontopathogenic bacteria as well as on the attachment and proliferation of osteoblasts and periodontal ligament fibroblasts.

Methods

Human alveolar osteoblasts (HAO) and periodontal ligament (PDL) fibroblasts were cultured on 3 concentrations of OCHS (2.5, 5 and 7.5 mg). Adhesion and proliferation were counted up to 48 h and mineralization was assayed after 1 and 2 weeks. Furthermore potential growth inhibitory activity on microorganisms associated with periodontal disease (e.g. Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans) as well as the influence of periodontopathogens and OCHS on the HAO and PDL fibroblasts counts were determined.

Results

More than a 2-fold increase in adherent HAO cells was observed at 4 h following application of OCHS when compared to the control group (p = 0.007 for 2.5 mg). Proliferation of HAO cells at 48 h was stimulated by moderate concentrations (2.5 mg; 5 mg) of OCHS (each p < 0.001), whereas a high concentration (7.5 mg) of OCHS was inhibitory (p = 0.009). Mineralization was observed only for HAO cells treated with OCHS. OCHS did not exert any positive effect on attachment or proliferation of PDL fibroblasts. Although OCHS did not have an antibacterial effect, it did positively influence attachment and proliferation of HAO cells and PDL fibroblasts in the presence of periodontopathogens.

Conclusions

The present data suggests that OCHS promotes osteoblast attachment, proliferation and mineralization in a concentration-dependent manner and results are maintained in the presence of periodontal pathogens.

【 授权许可】

   
2014 Eick et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Darveau RP: Periodontitis: a polymicrobial disruption of host homeostasis. Nat Rev Microbiol 2010, 8(7):481-490.
• [2]Report C: Periodontal diseases: pathogenesis and microbial factors. Ann Periodontol 1996, 1:926-932.
• [3]Socransky SS, Haffajee AD, Cugini MA, Smith C, Kent RL Jr: Microbial complexes in subgingival plaque. J Clin Periodontol 1998, 25(2):134-144.
• [4]Hernandez M, Dutzan N, Garcia-Sesnich J, Abusleme L, Dezerega A, Silva N, Gonzalez FE, Vernal R, Sorsa T, Gamonal J: Host-pathogen interactions in progressive chronic periodontitis. J Dent Res 2011, 90(10):1164-1170.
• [5]Lindhe J, Westfelt E, Nyman S, Socransky SS, Haffajee AD: Long-term effect of surgical/non-surgical treatment of periodontal disease. J Clin Periodontol 1984, 11(7):448-458.
• [6]Wikesjo UM, Selvig KA: Periodontal wound healing and regeneration. Periodontol 2000 1999, 19:21-39.
• [7]Tu YK, Needleman I, Chambrone L, Lu HK, Faggion CM Jr: A Bayesian network meta-analysis on comparisons of enamel matrix derivatives, guided tissue regeneration and their combination therapies. J Clin Periodontol 2012, 39(3):303-314.
• [8]Newman SA, Coscia SA, Jotwani R, Iacono VJ, Cutler CW: Effects of enamel matrix derivative on Porphyromonas gingivalis. J Periodontol 2003, 74(8):1191-1195.
• [9]Kasaj A, Willershausen B, Junker R, Callaway A, Krahn U, Kraft B, Pietsch M: Influence of different biomaterials on the viability of Aggregatibacter actinomycetemcomitans. Arch Oral Biol 2011, 56(9):917-923.
• [10]Polat HB, Yeler H, Gumus C, Bulut HE, Kucuk D: Effect of oil-based calcium hydroxide (Osteoinductal) on distraction osteogenesis in rabbit mandible. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009, 107(6):e30-e36.
• [11]Schwarz F, Stratul SI, Herten M, Beck B, Becker J, Sculean A: Effect of an oily calcium hydroxide suspension (Osteoinductal) on healing of intrabony periodontal defects. A pilot study in dogs. Clin Oral Investig 2006, 10(1):29-34.
• [12]Kasaj A, Willershausen B, Berakdar M, Tekyatan H, Sculean A: Effect of an oily calcium hydroxide suspension on early wound healing after nonsurgical periodontal therapy. Clin Oral Investig 2006, 10(1):72-76.
• [13]Stratul SI, Schwarz F, Becker J, Willershausen B, Sculean A: Healing of intrabony defects following treatment with an oily calcium hydroxide suspension (Osteoinductal). A controlled clinical study. Clin Oral Investig 2006, 10(1):55-60.
• [14]Kasaj A, Klein MO, Dupont J, Willershausen B, Krahn U, Gotz H, Zeiler J, Brullmann D, Duschner H: Early root surface colonization by human periodontal ligament fibroblasts following treatment with different biomaterials. Acta Odontol Scand 2013, 71(6):1579-1587.
• [15]Mohammadi Z, Dummer PM: Properties and applications of calcium hydroxide in endodontics and dental traumatology. Int Endod J 2011, 44(8):697-730.
• [16]Willershausen B, Willershausen I, Ross A, Velikonja S, Kasaj A, Blettner M: Retrospective study on direct pulp capping with calcium hydroxide. Quintessence Int 2011, 42(2):165-171.
• [17]Palmer NO, Ahmed M, Grieveson B: An investigation of current endodontic practice and training needs in primary care in the north west of England. Br Dent J 2009, 206(11):E22. discussion 584–585
• [18]Chala S, Abouqal R, Rida S: Apexification of immature teeth with calcium hydroxide or mineral trioxide aggregate: systematic review and meta-analysis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011, 112(4):e36-e42.
• [19]Higashi T, Okamoto H: Characteristics and effects of calcified degenerative zones on the formation of hard tissue barriers in amputated canine dental pulp. J Endod 1996, 22(4):168-172.
• [20]Stavropoulos A, Geenen C, Nyengaard JR, Karring T, Sculean A: Oily calcium hydroxide suspension (Osteoinductal) used as an adjunct to guided bone regeneration: an experimental study in rats. Clin Oral Implants Res 2007, 18(6):761-767.
• [21]Kohal RJ, Hurzeler MB, Schneider SR, Riede UN, Caffesse RG: The effect of a calcium hydroxide paste on wound healing and osseointegration of dental implants. A pilot study in beagle dogs. Clin Oral Implants Res 1997, 8(5):375-385.
• [22]Busenlechner D, Tangl S, Mair B, Fugger G, Gruber R, Redl H, Watzek G: Simultaneous in vivo comparison of bone substitutes in a guided bone regeneration model. Biomaterials 2008, 29(22):3195-3200.
• [23]Aparna S, Setty S, Thakur S: Oily calcium hydroxide suspension in the treatment of infrabony periodontal defects: a randomized controlled clinical trial. Quintessence Int 2011, 42(10):835-842.
• [24]Feres M, Haffajee AD, Allard K, Som S, Goodson JM, Socransky SS: Antibiotic resistance of subgingival species during and after antibiotic therapy. J Clin Periodontol 2002, 29(8):724-735.
• [25]Bennett JH, Carter DH, Alavi AL, Beresford JN, Walsh S: Patterns of integrin expression in a human mandibular explant model of osteoblast differentiation. Arch Oral Biol 2001, 46(3):229-238.
• [26]Miron RJ, Bosshardt DD, Zhang Y, Buser D, Sculean A: Gene array of primary human osteoblasts exposed to enamel matrix derivative in combination with a natural bone mineral. Clin Oral Investig 2012, 17(2):405-410.
• [27]Miron RJ, Bosshardt DD, Hedbom E, Zhang Y, Haenni B, Buser D, Sculean A: Adsorption of enamel matrix proteins to a bovine-derived bone grafting material and its regulation of cell adhesion, proliferation, and differentiation. J Periodontol 2012, 83(7):936-947.
• [28]Ferguson JW, Hatton JF, Gillespie MJ: Effectiveness of intracanal irrigants and medications against the yeast Candida albicans. J Endod 2002, 28(2):68-71.
• [29]Rosa OP, Torres SA, Ferreira CM, Ferreira FB: In vitro effect of intracanal medicaments on strict anaerobes by means of the broth dilution method. Pesqui Odontol Bras 2002, 16(1):31-36.
• [30]de Souza CA, Teles RP, Souto R, Chaves MA, Colombo AP: Endodontic therapy associated with calcium hydroxide as an intracanal dressing: microbiologic evaluation by the checkerboard DNA-DNA hybridization technique. J Endod 2005, 31(2):79-83.
• [31]Sathorn C, Parashos P, Messer H: Antibacterial efficacy of calcium hydroxide intracanal dressing: a systematic review and meta-analysis. Int Endod J 2007, 40(1):2-10.
• [32]da Silva RA, Leonardo MR, da Silva LA, de Castro LM, Rosa AL, de Oliveira PT: Effects of the association between a calcium hydroxide paste and 0.4% chlorhexidine on the development of the osteogenic phenotype in vitro. J Endod 2008, 34(12):1485-1489.
• [33]Wang S, Sasaki Y, Ogata Y: Calcium hydroxide regulates bone sialoprotein gene transcription in human osteoblast-like Saos2 cells. J Oral Sci 2011, 53(1):77-86.
• [34]Franceschi RT, Ge C, Xiao G, Roca H, Jiang D: Transcriptional regulation of osteoblasts. Ann N Y Acad Sci 2007, 1116:196-207.
• [35]Ganss B, Kim RH, Sodek J: Bone sialoprotein. Crit Rev Oral Biol Med 1999, 10(1):79-98.
• [36]Kasaj A, Willershausen B, Jewszyk N, Schmidt M: Effect of an oily calcium hydroxide suspension (Osteoinductal) on human periodontal fibroblasts. An in vitro study. Eur J Med Res 2007, 12(6):268-272.
• [37]Potempa J, Pike R, Travis J: Titration and mapping of the active site of cysteine proteinases from Porphyromonas gingivalis (gingipains) using peptidyl chloromethanes. Biol Chem 1997, 378(3–4):223-230.
• [38]Kato T, Tsuda T, Inaba H, Kawai S, Okahashi N, Shibata Y, Abiko Y, Amano A: Porphyromonas gingivalis gingipains cause G(1) arrest in osteoblastic/stromal cells. Oral Microbiol Immunol 2008, 23(2):158-164.
• [39]Yamamoto S, Mogi M, Kinpara K, Ishihara Y, Ueda N, Amano K, Nishihara T, Noguchi T, Togari A: Anti-proliferative capsular-like polysaccharide antigen from Actinobacillus actinomycetemcomitans induces apoptotic cell death in mouse osteoblastic MC3T3-E1 cells. J Dent Res 1999, 78(6):1230-1237.
• [40]Li X, Zhou L, Takai H, Sasaki Y, Mezawa M, Li Z, Wang Z, Yang L, Wang S, Matsumura H, et al.: Aggregatibacter actinomycetemcomitans lipopolysaccharide regulates bone sialoprotein gene transcription. J Cell Biochem 2012, 113(9):2822-2834.