Pascon EA  Spangberg LS   In vitro cytotoxicity of root canal filling materials: 1. Gutta-    percha.    In: J Endod (1990 Sep) 16(9):429-33    ISSN: 0099-2399   

Gutta-percha (GP) has been the most widely used root canal filling    material because of its well-known low toxicity. The inertness of GP,    however, was challenged recently. The purpose of this study was to    evaluate the toxicity of marketed endodontic GP using the    radiochromium release test. …The raw materials and barium sulfate were not toxic, whereas zinc oxide and zinc ions showed marked toxicity. All GP points tested were toxic at longer observation periods, and the toxicity was attributed to leakage of zinc ions into the fluids.

Takahara K  Onodera A  Matsumoto K   Toxicity of root canal sealers on rat bone cells in primary culture. Endod Dent Traumatol (1990 Oct) 6(5):200-7

The cytotoxic effects on cultured rat bone cells of newly-developed   root canal sealers and commercially available sealers were compared.   Various root canal sealers were applied to cultured bone cells   obtained from rat calvaria by the enzyme digestion method.   Measurement of [3H]-thymidine incorporation, alkaline phosphatase   activity, and calcium release were performed after 24 and 48 h. No   significant difference was found in cellular DNA synthesis and   alkaline phosphatase activity between cells exposed to New B-1, New B- 5 and controls after exposure for 24 and 48 h. Cells in contact with   Tubliseal, Diaket and AH-26 demonstrated a significant difference   from controls in DNA synthesis and alkaline phosphatase activity.   Calcium release at 24 h was significantly different in the cells   treated with New B-1 and New B-5 than in controls. No appreciable   difference was found, however, between New B-6, Sealapex and   controls. At 48 h, cells treated with New B-1, New B-5 and New B-6   showed differed significantly difference from controls, but the cells   exposed to Sealapex did not. The newly-developed root canal sealers   had lower toxicity in vitro than five types of commercially available   root canal sealers.

Arenholt-Bindslev D  Horsted-Bindslev P   A simple model for evaluating relative toxicity of root filling   materials in cultures of human oral fibroblasts. Endod Dent Traumatol (1989 Oct) 5(5):219-26

 Standardized test tubes filled with freshly mixed root filling   materials (AH26, CRCS, N2, Kloroperka NO, ZOE cement and 2   experimental cements, ECI and ECII) were transferred into tissue   culture flasks. Normal human oral fibroblasts were seeded in the   flasks. Morphological cell changes were studied up to 15 days after   seeding. The size of cell-free zones around the test tubes and the   total cell number per culture flask were calculated after 5, 10 and   15 days. The findings showed N2 cement to be by far the most toxic material at all observation periods, whereas no toxic reactions could be seen in relation to tubes filled with Kloroperka NO. Compared with the 5-day observation period, some cell recovery was observed around test tubes with AH26 and ECII, whereas almost full cell recovery was found around test tubes with CRCS, ZOE and ECI. It was concluded that the present model, which allows long-term observations of human cellular reactions to dental materials, can be used as a simple and relatively cheap screening test for initial toxicity testing of   dental materials.

Pissiotis E  Spangberg LS   Toxicity of Pulpispad using four different cell types. Int Endod J (1991 Sep) 24(5):249-57

The cytotoxic effect of a zinc oxide-eugenol-based paste (Pulpispad)   was evaluated in vitro after setting for 1 day and 1 week. Target   cells were L929 cells, gingival, periodontal ligament and pulpal fibroblasts. The material was incubated with the cells for 4 and 24 hours, and its toxicity was evaluated with the 51Cr-release method.   Pulpispad was highly cytotoxic to all cell lines even after setting for 1 week. The use of Pulpispad is not recommended for future clinical application. The various responses among the four cell lines indicated that diploid cell lines can, under certain circumstances,   be less sensitive than aneuploid cell lines. It is therefore suggested that in the evaluation of biomaterials the choice of cell lines should be carefully considered, as they can display varying   sensitivities.

Lewis BB  Chestner SB   Formaldehyde In Dentistry: A Review Of Mutagenic And Carcinogenic   Potential J Am Dent Assoc (1981) 103(3):429-434

 A review of the carcinogenic and mutagenic potential of formaldehyde   products used in dentistry is presented. Dentists performing   endodontic therapy may use formocresol and formaldehyde paste. Tissue   contact results either directly or by contact with the gaseous   formaldehyde phase. While various short-term tests have indicated the   mutagenicity of formaldehyde, they are not conclusive for the   mammalian species. Clinical studies involving chronic topical   application of formalin have demonstrated the induction of   leukoplakia and lesions resembling carcinoma in situ. The respiratory   tract may be the area for the greatest risk of the development of   tumors. Animal studies demonstrate that formaldehyde may affect   reproduction potential. Formaldehyde should be considered a potential   carcinogen in humans. Formaldehyde poses problems to systemic health   via ingestion routes, interaction in air with other aldehydes, and in   final breakdown products of formalin in the body. If formaldehyde is   clearly not necessary, why is it used at all, particularly in light   of its deleterious effects. (115 Refs)

Stea S  Savarino L  Ciapetti G  Cenni E  Stea S  Trotta F  Morozzi G     Pizzoferrato A   Mutagenic potential of root canal sealers: evaluation through Ames   testing. J Biomed Mater Res (1994 Mar) 28(3):319-28

The mutagenic potential of 12 commercially available dental cements   and of two ‘pure substances’ (zinc oxide and eugenol) used in root   canal filling were examined. The cements were prepared according to   the manufacturers’ indications and set for defined times. Ames tests   were performed in their extracts by using Salmonella typhimurium   strains TA 98 and TA 100. The results showed that most cements   present strong bactericidal activity that disappears or decreases   remarkably in time. One of the tested cements showed mutagenicity   with both Salmonella strains. Two cements yielded doubtful results.   The remaining cements and the two ‘pure substances’ showed no   mutagenic potential. The authors conclude that it is convenient to   examine endodontic cements with the Ames test and to eliminate those   that present mutagenicity in time.

Al-Nazhan S  Spangberg L   Morphological cell changes due to chemical toxicity of a dental   material: an electron microscopic study on human periodontal ligament   fibroblasts and L929 cells. J Endod (1990 Mar) 16(3):129-34

New endodontic materials with polymer bases may be more difficult to   evaluate in cell cultures in vitro than conventional zinc oxide-   eugenol cements. In order to study the morphological changes taking   place in cells exposed to such materials, L929 cells and human   periodontal fibroblasts were observed using scanning electron   microscopic and transmission electron microscopic techniques. The   morphological changes of the cells were correlated to the   quantitative results observed simultaneously in cytotoxicity studies   using the radiochromium release method. Results showed there was a   relationship between the chromium release and the degree of   individual cell damage. The periodontal ligament fibroblasts were   more resistant to this kind of chemical injury than the L929 cells.   Consequently, it may be proper to use periodontally derived cells for   the study of cytotoxic mechanisms of polymer endodontic filling   materials.

Safavi KE  Spangberg LS  Costa NS Jr  Sapounas G   An in vitro method for longitudinal evaluation of toxicity of   endodontic sealers. J Endod (1989 Oct) 15(10):484-6

An in vitro method for longitudinal evaluation of root canal sealers   was developed and applied. A newly introduced cell culture chamber   was used to evaluate the cytotoxic effects of test samples   immediately after mixing and for an extended period of time   thereafter. A ranking of the test materials, based on their   cytotoxicity, was allowed by the method.

Chutich MJ  Kaminski EJ  Miller DA  Lautenschlager EP   Risk assessment of the toxicity of solvents of gutta-percha used in   endodontic retreatment. J Endod (1998 Apr) 24(4):213-6

Three randomly assigned groups of single-canaled extracted teeth   obturated with gutta-percha were retreated using controlled   application of one of three organic solvents: chloroform, xylene, or   halothane. Two additional groups of teeth served as positive and   negative controls. Residual volume of solvent expressed through the   apical foramen during retreatment was determined by the difference of   pretreatment and posttreatment weights of hermetically sealed   receptacles attached to the root surface of the teeth. Results   indicate that the amount of solvent that has been determined to have   leached out through the apical foramen is several orders of magnitude   below the permissible toxic dose. Thus, it is proposed that the use   of any of the aforementioned solvents used in the retreatment of root   canals would pose negligible risk to the patient.

Mittal M  Chandra S  Chandra S   Comparative tissue toxicity evaluation of four endodontic sealers. Endod (1995 Dec) 21(12):622-4

The tissue toxicity of zinc oxide-eugenol, Tubli seal, Sealapex, and   Endoflas F.S. was investigated by injecting them into the   subcutaneous connective tissue of the dorsal surface of rats and   studying the tissue response histologically. Animals were killed   after time intervals of 48 h, 7 days, 14 days, 1 month, and 3 months;   tissue sections were taken from the injection sites. Histological   examination of the tissue sections revealed that all of the sealers   caused some inflammation that decreased with time, except in the case   of zinc oxide-eugenol where it increased from the 48th hour to the   7th day and after that showed a decreasing trend. Overall, Sealapex   showed the least inflammatory reaction compared with other sealers   used, because it showed moderate inflammation at 48 h that became   mild in later periods. Zinc oxide-eugenol, Tubli seal, and Endoflas   F.S. were severely toxic at 48 h and 7 days. This toxicity decreased   gradually in later time periods. No inflammatory reaction was seen at   3 months with any of the sealers used.

Barbosa SV  Burkard DH  Spangberg LS   Cytotoxic effects of gutta-percha solvents. Endod (1994 Jan) 20(1):6-8

Cytotoxicity of commonly used gutta-percha solvents was evaluated.   Gutta-percha dissolved by chloroform, halothane, or turpentine was   evaluated with the radiochromium release method using L929 mouse   fibroblast cells. All solvents were toxic. Turpentine was most toxic   followed by halothane and chloroform, which caused similar levels of   cell injury.

Helseth DL Jr  Tolwin TM  Kaminski EJ  Osetek EM   Incomplete polymerization of Cavalite with the use of recommended   photopolymerization times: a warning of possible cytotoxic effects.Oral Surg Oral Med Oral Pathol (1989 Aug) 68(2):223-5

As part of a study of the suitability of new materials for use as a   retrofilling material, we examined the polymerization properties of   Cavalite, a light-cured, hydroxyapatite and glass ionomer-containing   cavity liner. By varying the time of photopolymerization, it was   found that polymerization for 20 to 30 seconds according to the   manufacturer’s recommendations is not sufficient to ensure complete   polymerization. The implications of this incomplete polymerization   are discussed in terms of possible cytotoxic effects on tissues   exposed to unpolymerized Cavalite, both when used in retrofilling   situations and as a deep cavity liner.

Yesilsoy C  Koren LZ  Morse DR  Kobayashi C   A comparative tissue toxicity evaluation of established and newer   root canal sealers.Oral Surg Oral Med Oral Pathol (1988 Apr) 65(4):459-67

Grossman’s sealer, eucapercha, Endo-Fill, CRCS, Sealapex, Hypocal,   and sterile saline solution (0.3 ml of each) were injected into  specific dorsal subdermal tissue sites of 12 guinea pigs. The animals were killed after 6 days, 15 days, and 80 days (four per time   period). Analysis of tissue response showed that, overall, Sealapex   and Endo-Fill had less severe inflammatory reactions than any of the other test materials. Grossman’s sealer, CRCS, and Hypocal showed   principally severe inflammatory responses at both 6 and 15 days, but   mild reactions at 80 days. Overall, eucapercha showed less severe   inflammatory responses than Grossman’s sealer, CRCS, and Hypocal.   Diffuse calcification was induced by the three calcium hydroxide preparations (CRCS, Sealapex, and Hypocal). Eucapercha and Endo-Fill   had minute local areas of calcification. Both Grossman’s sealer and CRCS did not have overall favorable histologic reactions; however,  Grossman’s sealer and CRCS have been used successfully clinically.   Further clinical studies are needed.

Wright KJ  Barbosa SV  Araki K  Spangberg LS   In vitro antimicrobial and cytotoxic effects of Kri 1 paste and zinc   oxide-eugenol used in primary tooth pulpectomies. Pediatr Dent (1994 Mar-Apr) 16(2):102-6

The antimicrobial and cytotoxic effects of Kri 1 paste, an iodoform-   based primary tooth filling material, were compared with zinc oxide-   eugenol (ZOE), using in vitro techniques. Antimicrobial evaluation   involved measuring inhibition zones of Streptococcus faecalis on   brain heart agar. Cytotoxicity evaluation involved direct cell-   medicament contact experiments of 4-hr and 24-hr duration using fresh   and set medicaments, and indirect cell-medicament contact experiments   of 24-hr duration using fresh and set medicaments. ZOE produced a   greater zone of bacterial inhibition than Kri 1 paste. Kri 1 paste   cytotoxicity remained high regardless of the amount of setting time   in the 4-hr direct contact experiment, while ZOE cytotoxicity   decreased with setting time. Both Kri 1 paste and ZOE had high   cytotoxicity regardless of setting time in the 24-hr direct cell-   medicament contact test. ZOE cytotoxicity decreased to control levels   after only 1 day of setting in the indirect contact experiments,   compared with greater than 7 days for Kri 1 paste. The results   suggest ZOE has better antimicrobial activity than Kri 1 paste. ZOE   also has lower cytotoxicity, although prolonged cell-medicament   contact may result in both medicaments having similarly high   cytotoxicity.

Lambrecht JT  Panzer G   [The toxicity of root-canal filling materials in primary osteoclast   cell cultures (see comments)]Die Toxizitat von Wurzelfullmaterialien in primaren Osteoklasten-   Zellkulturen.Schweiz Monatsschr Zahnmed (1995) 105(7):899-906

Endodontic root-filling material is brought into direct contact with   apical tissues when finishing endodontic therapy. Endodontic two-   component materials develop cytotoxic effects during the phase, as   shown in different in-vitro studies. Insufficient specificity of   these tests could be eliminated by using bone cell cultures. The   toxicity of seven endodontic root-filling materials towards   osteoclasts in primary cell culture were investigated. Osteoclasts   initially reacted to freshly mixed endodontic materials by losing   their physiological properties (surface adherence), longer exposition   on the toxic agents led to cell lysis. To quantify these light-   microscopical phenomena, the content of lactate dehydrogenase (LDH)   in the adherent cells as well as in the medium of the cultures were   determined photometrically. All the examined two-component materials   induced a distinct loss of intracellular LDH activity with a   proportional uprise in the media. Calibration showed a positive   correlation of LDH content and cell number. The results could be   interpreted as the survival rate of osteoclasts after incubation with   endodontic filling materials. All tested materials–except for gutta-   percha–showed a distinct toxicity towards osteoclasts in primary   cell culture during the first 24 hours. Osteoclasts proved to be   sensitive indicators for cell-adverse toxicity during the in-vitro   tests of endodontic root-filling materials.