DPSCs were treated with MTA, Ca(OH)2, Biodentine or Emdogain. capping. One of these limitations is that we have relied on the MTT assay to examine cell viability and not fully examined the cellular toxicity from apoptosis and necrotic cell death pathways, that remain to be studied. Cell viability assays are often used to determine the effects of test compound on cell proliferation or cytotoxicity. J Endod 36(5): 806-813. defect vacuole J Dent Res 92(7 Suppl): 16S-22S. 
velocity Dominguez MS, Witherspoon DE, Gutmann JL, Opperman LA. In the item Other Pulpal Findings, MTA as direct pulp capping materials. J Dent Res 74(9): 1602-1606. cDNA synthesis was performed by using high capacity cDNA reverse transcription kit according to manufacturers instruction (Applied Biosystems, Thermo Fisher Scientific, USA). respectively) according to Vural UK et al. hydration reaction, and polycarboxylate reduces the amount of be affected by the particle size and distribution as well as by the Hammarstrm L, Heijl L, Gestrelius S. Periodontal regeneration in a buccal dehiscence model in monkeys after application of enamel matrix proteins. M, Parirokh M (2011) A comparative study on dental pulp response to Stem Cells Int. Int Dent J 62(1): 33-39. procedure is not recommended [1]. Dental pulp was extirpated and DPSCs were isolated and characterized by flowcytometry. DPSCs are characterized by expression of common differentiation markers including CD29, CD44, CD73, CD90, CD105, CD146 and absence of CD14, CD34, CD45 [54]. The use of dental pulp stem cells (DPSCs) in research to understand the various mechanisms, gained momentum in many dental research laboratories. The solution was transferred to 96-well plate at 100l/well and optical density was read at 570nm by a spectrophotometric Microplate Reader (SpectroStar Nano, BMG Lab). The pulp capping materials form a protective layer over the nearly exposed or exposed vital pulp in direct and indirect pulp capping or in pulpotomy procedures [50]. pulp-capping materials for permanent teeth. Analyzing the role of different biomaterials in the osteogenic gene expression and cell viability of stem cells is important for elucidating the molecular mechanisms underlying tissue engineering and regenerative medicine. Calcium Hydroxide. capping systematic 2016;24(5):4816. inconvenient to both dentist and patient, because it requires direct Asgary S, Eghbal MJ, Parirokh M, Ghanavati F, Rahimi H. A comparative study of histologic response to different pulp capping materials and a novel endodontic cement. 2012;3(1):2041731412467998. Schmalz G, Schweikl H, Esch J, Hiller KA (1996) Evaluation of a dentin avoided through the conservative approach of direct pulp capping. with Dycal than with Propolis and MTA on the 15th as well as on the Further studies to assess the inflammatory mediators resulting from interaction of DPSCs with pulp capping materials may explain cytotoxicity of these materials. DPSCs were cultured in -MEM growth medium and characterized by flow cytometry using CD90 and CD45 surface markers. After solubilization, chloroform was added and the upper phase containing the RNA was separated to which equal volume 70% ethanol was added. and its ability to aid in the release of bioactive dentin matrix Therefore, the aims of this study was to evaluate and compare the in vitro osteogenic, odontogenic and angiogenic effects of mineral trioxide aggregate (MTA), calcium hydroxide [Ca(OH)2], Biodentine and Emdogain on dental pulp stem cells (DPSCs) and examine the effects of the tested materials on cell viability. Several angiogenesis markers such as VEGF have been investigated. 
[59] have found that Emdogain enhanced proliferation of DPSCs in osteogenic induction medium and Arajo et al. Inflammation and General State of the Pulp (p>0.05), Calcium Firm necrosis causes many tooth extractions and root canal treatments could have been 349-353. capping: long-term results. The effect of different pulp capping materials on cell viability of dental pulp stem cell after 3days. Gandolfi MG, Ciapetti G, Taddei P, Perut F, Tinti A, Cardoso MV, et al. MTA is primarily calcium oxide in the form of tricalcium silicate, They found the reparative dentin formed in the calcium hydroxide, Kemoun P, Gronthos S, Snead ML, Rue J, Courtois B, Vaysse F, et al. 2003;74(8):11915. MTA, Ca(OH)2, Biodentine and Emdogain increased DSPP expression considerably on day 7 (Fig. effectiveness of mineral trioxide aggregate (mta) and calcium hydroxide Nair PN et al. area to facilitate both the formation of protective barrier and the 
bridge; furthermore, significantly greater immunostaining was The relative gene expression by real time PCR was normalized against the internal control gene (GAPDH) and relative to the untreated control. dental dam isolation of the tooth; (5) maintenance of adequate seal Cox CF, Sbay RK, Ostro E, Suzuki S, Suzuki SH (1996) Tunnel defects Baume L, Holz J (1981) Long-term clinical assessment of direct pulp the odontoblast-like cells and pulp fibroblasts beneath the dentin The clinical and historical findings Hydroxide was the purpose of Swarup SJ et al. Nakamura Y, Slaby I, Matsumoto K, Ritchie HH, Lyngstadaas SP. The cells were adherent to plastic plate and expressed CD90 stem cell marker and lack expression of the hematopoietic lineage marker CD45. when used as a direct pulp capping agent: an in vivo study. J Endod. Calcium Hydroxide were evaluated by Iwamoto C et al. Biomaterials 27(14): 2865-2873. (2007) Ramy Emara isolated the dental pulp, prepared the dental materials and helped in the development of the main research idea. J Dent Educ 67(6): 678-689. In mineral trioxide aggregate (WMTA, GMTA) and Dycal as a pulp it can prevent the diffusion of potentially toxic materials through formation, less inflammatory response and less necrosis of pulpal Am J Dent 13(2): 81-87. The cumulative success rate of both materials was not Histological evaluation of mineral trioxide aggregate and calcium However, Ca(OH)2 was the only capping material that reduced VEGF expression on day 7 and day 14. After 7 and 14days of treatment, DPSCs were rinsed three times in PBS and TRIzol Reagent (Invitrogen, Thermo Fisher Scientific, USA) was added directly to the attached cell to lyse them. barrier test by cytotoxicity testing of various dental cements. eugenol in human osteoblastic cells in vitro. exposure. 2007;33:12314. Parolia A stimulate pulp repair [10,21,22]. Torabinejad M, Hong CU, McDonald F, Pitt Ford TR (1995) Physical and than teeth treated with Dycal on the 45th day. Bismuth oxide is added Vital pulp therapy maintains dental pulp vitality and preserves teeth [32]. Enamel matrix derivative (EMD, Emdogain) is composed of a mixture of hydrophobic enamel matrix proteins derived from 6-month-old porcine tooth buds containing amelogenin, enamelin, tuftelin, amelin, and ameloblastin [11, 52], in a propylene glycol alginate (PGA). Winer J, Jung CK, Shackel I, Williams PM. Dent Traumatol Bio Zhao Y et al. non-significant. direct pulp capping material, showed higher success rate than dycal. [53], We would like to thank Deanship of scientific research at Umm Al-Qura University, Makkah, KSA for funding this project (Code number 43509026) to Dr. Abdelrahman Youssef. J Endo trial. ultrastructural and quantitative investigations on the response of In endodontic regeneration various studies have examined the effect of biomaterials on vascularization and angiogenesis [7, 24, 25]. 2010;36(11):183943. J Endod. Angiogenic factor VEGF expression was upregulated by Biodentine and Emdogain on day 7 (Fig. The mechanisms of observed cytotoxicity are not clearly understood. pulp-capping with MTA in two visits: application of MTA in the first of the strictly anaerobic bacteria. Calcium Hydroxide group. the second to sixth week after operation (P<0.05). On J Dent & Oral Health. Emdogain Gel 30mg/ml (BIORA AB/Straumann, Switzerland) was diluted with alpha-modified Eagle medium (-MEM) to a final working concentration of 100g/ml. Mente J, Hufnagel S, Leo M, Michel A, Gehrig H, et al. For for direct pulp capping is questionable, however. J Endod 41(8): 1234-1240. agents on pulp cell migration, proliferation, and differentiation. the effectiveness of MTA and a conventional Calcium Hydroxide comparative histological analysis of human pulp following direct pulp Results shown here may be applied in future studies to modify the surface of the materials to promote better cell viability and sprouting of cells in tissue regeneration protocols. DPSCs were treated with MTA, Ca(OH)2, Biodentine or Emdogain. capping. involved tomographic evaluations of reparative dentin bridge 2016;20(2):23746. hard tissue barrier that had numerous tunnel defects. capped with white ProRoot Mineral Trioxide Aggregate (WMTA) endodontic use. process and more predictable hard tissue barrier formation. Nawras Maher Mostafa1 and Shady Ahmed Moussa2*. Calcium carbonate J Am Dent Assoc 139(3): 305- to Calcium Hydroxide make it the material of choice in direct pulp approximately the same as 24 grams of calcium hydroxide base/ 4a). Hydroxide. necrosis was more with Nano-HA and calcium Hydroxide which 1995;21:228. Youssef, AR., Emara, R., Taher, M.M. The percentage of stem cell viability was compared to control (100%). Matsuo T, Nakanishi T, Shimizu H, Ebisu S (1996) A clinical study of the two materials. with white MTA was also described in case reports in endodontic Dental stem cell and dental tissue regeneration. Int Endod J 46(2): 137-144. Oral Health, Zagazig University, Egypt & Consultant of Pediatric Dentistry and Dental catalyst paste, making MTA much less cost effective per use [10]. Mustafa S, Jamil, Sajjid I, Munir B, Kouser R (2016) Comparison of the products showed comparable results at day 60. 
There are three causes J Clin Periodontol. Nakamura Y, Hammarstrom L, Matsumoto K, Lyngstadaas SP. Biomed Res Int. https://doi.org/10.1186/s12903-019-0827-0, DOI: https://doi.org/10.1186/s12903-019-0827-0. [50] to assess the effectiveness Stem cell properties of human dental pulp stem cells. influence on the outcome of direct pulp capping (p>0.05). the mouth. DPSCs at the third passage were cultured in -MEM growth medium at 2105/well into 6-well plates (Costar, Corning Life Sciences, USA). 2014;42(1):539. 1BDS, MSc, Specialist of Restorative and Advanced Aesthetic Dentistry, PHCC, Qatar. 2011;32(30):737588. Bio dentine (Septodont, Saint Maur des Fosses, France), and Single or Calcium Hydroxide and they found no statistically significant Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, et al. The method of DPSCs isolation was first reported in 2000 by Gronthos et al. Growth factors as key mediators. Similar results were reported by other studies, for ex., osteogenesis markers were upregulated by Emdogain [26, 30, 59], MTA and Biodentine [9]. There are a number of We used the MTT assay to compare the cytotoxic effect of MTA, Ca(OH)2, Biodentine and Emdogain on DPSC. the event of exposure in vital pulp, direct pulp capping, pulpotomy ZOE also demonstrates high Untreated cells were used as control. The initial effect of Calcium Hydroxide applied to exposed pulp (2nd edn), Williams & Wilkins, Philadelphia, USA, pp. avoided through the conservative approach of direct pulp-capping 197. Costa CA, Vaerten MA, Edwards CA, Hanks CT (1999) Cytotoxic effects of None of the clinical variables had a considerable statistically different (P=0.282). formed in the Single Bond Universal group in terms of thickness Clin Exp Med 8(10): 17055-17060. pH causes irritation of the pulp tissue, which stimulates repair via MTA is more effective and superior comparing the Calcium Hydroxide as a Although DPSCs isolated from inflamed pulp have altered stem cell properties, they hold the potential to regenerate tissues [4]. Histogram representing the cell count (Y-axis) and fluorescence intensity on X-axis. approximately 2 hours and 45 minutes. Regarding to the performance and success of the MTA compared using as a direct pulp capping compared to Calcium Hydroxide. However, both MTA and Calcium Hydroxide were clinically Propolis and MTA showed bridge formation in more teeth, and pulp inflammation associated with various restorative materials. treatments. on the erosion properties of calcium hydroxide cements. They found One hundred and fifty-two teeth of 172-capped teeth influence of mineral trioxide aggregate and calcium hydroxide on dental The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. eugenol release, it must be kept in mind that eugenol release drops Complete dentinal The effect of pulp capping materials on cell viability of DPSCs was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction assay according to manufacturer instructions (Abcam, UK). pulpotomy evaluating clinical pulp conditions, the most important test is pulp Another frequently used assay is the luminogenic ATP assay in which the amount of ATP is directly proportional to the number of cells. Provided by the Springer Nature SharedIt content-sharing initiative. J Endod. (2014) Treatment J Mater Sci Mater Med. has no inherent adhesive qualities and provides a poor seal [1,10] times (from 10 to 12 min). dentinogenic process and more predictable hard tissue barrier formation. Also, we would like to thank Dr. Ahmed Tarek, Dr. Ashraf Abdeljalil, and Mrs. Rania El-Nahas for technical help. applicable at the end of 12 months follow up period and there was bridge formation, an absence of inflammatory pulp response and layers of well-arranged odontoblasts and odontoblast-like cells Nair PN, Duncan HF, Pitt Ford TR, Luder HU (2008) Histological, Hydroxide showed a tendency towards presenting a higher Hydroxide group (P=0.006). Murray PE, Hafez AA, Windsor LJ, Smith AJ, Cox CF (2002) Comparison LPS. sialoprotein and heme oxygenase-1 in human dental pulp. 1997;24(9 Pt 2):66977. 3d). 1(2): 2018. However, further future research is required to assess the clinical outcomes and compare it with the in vitro findings. At the sixth week, there was The authors declare that they have no competing interests. Moreover, it may increase the 2003;29(3):1769. [Updated 2016. asymptomatic pulp; (2) controlled bleeding at the exposure site; 2018. https://doi.org/10.1007/s11684-018-0628-x. should show normal apical tissue. of Glass ionomers ability to chemically bond to tooth structure, reduced with time. VEGF stimulates the endothelial cells close to microvessels to proliferate, migrate and change their pattern of gene expression [12]. Additional studies are needed to assess the clinical outcomes and compare it with the in vitro findings. trioxide aggregate: a review from the biological and physicochemical J Endod 40(3): 436-440. risk of bacterial contamination [10,30,31]. contamination following dentinbridging beneath hard-setting calcium bioactive dentin matrix molecules as liners in deep cavities prepared in Our data are in agreement with Bortoluzzi et al. (1985) The Biodentine is used for endodontic repair and pulp capping [38] and has good biocompatibility with DPSCs [9]. nano hydroxyapatite, mineral trioxide aggregate and calcium hydroxide of ZOE in excluding bacteria is reduced the longer it is in place in maintaining long-term tooth vitality. 2012;347(2):41927. Dentin bridge was not observed in Dycal group for the dentin to the pulp. and are subject to dissolution over time [10-19] Calcium Hydroxide Regenerative medicine is the process of restoring or regenerating human cells, tissues or organs for therapeutic applications [40]. Hilton TJ, Ferracane JL, Mancl L; Northwest Practice-based Research 2BDS, MSc, DDS, MRACDS (DPH), Lecturer and consultant in Pediatric Dentistry and Oral Health Zagazig University, Egypt. If pulp exposure occurs during J Endod 40(11): 1746-1751. However, TheraCal decreased pulp fibroblast proliferation and induced proinflammatory IL-8 secretion by pulp fibroblasts [29]. cement, and glass-ionomer cement against three fibroblastic The mean thickness of the dentin bridges observed MTA provided better long term results more effective than Calcium Hydroxide in maintaining long-term vitality. 2012;56:63949. Pakistan Oral & Dental J 36(2): 319- However, the main drawbacks of MTA include long setting time, difficulty in removal after setting and its application must be in an area free from any infections [8, 10, 14, 15, 17]. OJDOH.MS.ID.000508. Real Time-PCR was performed on day 7 and 14 (Figs. The The study was done by Nowicka A et al. results were positive after a direct coverage of the pulp using Effects of mineral trioxide aggregate, calcium hydroxide, biodentine and Emdogain on osteogenesis, Odontogenesis, angiogenesis and cell viability of dental pulp stem cells, https://doi.org/10.1186/s12903-019-0827-0, https://doi.org/10.1007/s11684-018-0628-x, http://creativecommons.org/licenses/by/4.0/, http://creativecommons.org/publicdomain/zero/1.0/. In: Gilbert D, Friedrich O, editors. The cell viability was measured by MTT assay on day 3. and white MTA. FASEB J. Immunohistochemical characterization of rapid dentin formation induced by enamel matrix derivative. current dental adhesive systems on immortalized odontoblast cell line Our results suggest MTA, Biodentine and Emdogain exhibit similar attributes and may score better thanCa(OH)2. Camilleri J, Montesin FE, Di Silvio L, Pitt Ford TR (2005) The chemical Brown LF, Detmar M, Claffey K, Nagy JA, Feng D, Dvorak AM, Dvorak HF. There was non-significant range (P New York, NY: Humana Press; 2017. p. 1601. dentin bridges was the highest in the MTA group and the lowest in and P=0.002, respectively and the WMTA calcified bridge formed Its use The study of Leye Benoist F et al.
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