Home | About Us | Editorial Board | Current Issue | Archives | Search | Instructions | Subscription | Feedback | e-Alerts | Login 
Journal of Indian Society of Pedodontics and Preventive Dentistry Official publication of Indian Society of Pedodontics and Preventive Dentistry
 Users Online: 329  
  Print this page Email this page   Small font sizeDefault font sizeIncrease font size

  Table of Contents    
Year : 2016  |  Volume : 34  |  Issue : 3  |  Page : 269-272

An in vitro evaluation of cytotoxicity of curcumin against human dental pulp fibroblasts

1 Department of Pedodontics and Preventive Dentistry, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India
2 Department of Microbiology and Molecular Biology, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka, India
3 Department of Pedodontics and Preventive Dentistry, Bapuji Dental College and Hospital, Davangere, Karnataka, India

Date of Web Publication25-Jul-2016

Correspondence Address:
Praveenkumar S Mandrol
Department of Pedodontics and Preventive Dentistry, Mandal's NGH Institute of Dental Sciences and Research Centre, Belagavi, Karnataka
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0970-4388.186757

Rights and Permissions



Objective: The objective of this study was to evaluate the cytotoxicity of curcumin to primary dental pulp fibroblasts in vitro. Materials and Methods: Dental pulp fibroblasts from primary maxillary central incisors were cultured and used for cytotoxicity tests after the fourth passage. Ninety-five percent curcumin was diluted with dimethylsulfoxide to prepare 100%, 50%, and 25% concentrations. Each concentration of curcumin was added in triplicate into 96-well microtiter plate containing the fibroblast culture at 104/well. Cells without treatment served as a control group. The number of viable cells after 48 hrs incubation at 37°C in a humidified atmosphere of 5 % CO2 and 95 % air was determined by the 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The relative viability of pulp cells was expressed as color intensity of the number in the experimental wells relative to that of the control group. Absorbances were read at 492 nm on a microplate reader with a background subtraction at 620 nm. Results: Cell viability of primary dental pulp fibroblasts to 25%, 50%, and 100% curcumin concentration was 174%, 310%, and 317%, respectively. Conclusions: Curcumin promotes cell viability and induces proliferation of primary dental pulp fibroblasts and has the potential to be developed into an economical and reliable medicament for vital pulp therapy.

Keywords: Curcumin, cytotoxicity, pulp fibroblasts

How to cite this article:
Mandrol PS, Bhat K, Prabhakar A R. An in vitro evaluation of cytotoxicity of curcumin against human dental pulp fibroblasts. J Indian Soc Pedod Prev Dent 2016;34:269-72

How to cite this URL:
Mandrol PS, Bhat K, Prabhakar A R. An in vitro evaluation of cytotoxicity of curcumin against human dental pulp fibroblasts. J Indian Soc Pedod Prev Dent [serial online] 2016 [cited 2023 Jan 29];34:269-72. Available from: http://www.jisppd.com/text.asp?2016/34/3/269/186757

   Introduction Top

Management of the grossly carious primary molar is a common but sometimes challenging aspect of dental care for young children. Pulpotomy is performed in a primary tooth with extensive caries, but without evidence of radicular pathology when caries removal results in a carious or mechanical pulp exposure. The coronal pulp is amputated, and the remaining vital radicular pulp tissue surface is treated with a long-term clinically successful medicament.[1],[2] An optimum wound healing dressing or agent protects the wound tissue from bacterial infection, reduces inflammation, and induces cell proliferation to aid in the reconstruction of damaged tissue.[3] It would ideally also act as an anti-oxidant, as free radicals are considered the major cause of inflammation during wound healing process.[4] The objective is that radicular pulp should remain healthy without adverse clinical signs or symptoms such as sensitivity, pain or swelling, and also with no postoperative radiographic evidence of pathologic external or internal root resorption and no harm to succedaneous tooth [1],[5] so that the primary tooth fulfills its role in the dentition.

Traditional medicine is known to be fertile ground for the source of modern medicines. One medicine in that category is turmeric (Curcuma longa) and belongs to the ginger (Zingiberaceae) family. Components of turmeric are named curcuminoids, which include mainly curcumin (diferuloyl methane), demethoxycurcumin, and bisdemethoxycurcumin. Curcumin, a yellow phenolic pigment, is the most important fraction which is responsible for the biological activities of turmeric.[6] Extensive research on curcumin has demonstrated a wide spectrum of therapeutic actions such as anti-inflammatory, antibacterial, antiviral, antifungal, anti-diabetic, anti-coagulant, hepato-protective, anti-ulcer, hypo-tensive, and hypo-cholesteremic.[6],[7] How a single agent could exhibit all these effects is an enigma under intense analysis. Pharmacological safety and small cost make curcumin an attractive agent to investigate further.

Many types of biomaterials have been utilized for the restorative and endodontic treatment procedures. The biological compatibility of dental materials is of vital importance to avoid or limit pulp tissue irritation or degeneration. Cytotoxicity screening assays provide a measure of cell death caused by materials or their extracts.[8] Primary cell cultures derived from dental pulp, i.e., pulp fibroblasts, may arguably be more relevant for testing the biocompatibility of materials for use in restorative dentistry and endodontics. Moreover, pulp fibroblast is highly sensitive to toxic substances, indicating that pulp cells could be a sensitive barometer to reveal the possible adverse effects of dental materials.[9],[10] As the rationale for the development of new therapeutic materials is to enhance successful clinical applications, trials must be carried out to evaluate their cytotoxicity. Hence, the aim of this study was to evaluate the cytotoxicity of curcumin on cultured human primary pulp fibroblasts.

   Materials and Methods Top

Source of explants

Human dental pulp fibroblasts were obtained from healthy patients presented to the Department of Pedodontics and Preventive Dentistry for orthodontic extractions. Included teeth were primary maxillary incisors with more than half root resorbed, which were devoid of any caries, restorations, and periodontal disease. Teeth were extracted aseptically after informed consent of parents. After extraction of teeth, pulp tissue was carefully extirpated from the root canal with sterile broach and sharp spoon excavator and was transported to the laboratory in Dulbecco's Modified Eagle's Medium (Hi-Media Laboratories, Mumbai, India) for fibroblast culture.

Fibroblast isolation and harvesting from dental pulp

Pulp tissue was placed in a sterile  Petri dish More Details containing a solution of 3 mg/ml collagenase and 4 mg/ml dispase for 1 h, till the tissue underwent enzymatic dissociation. Small pieces of pulp tissues were removed using a micropipette, re-suspended in 5–10 ml phosphate buffered saline, and centrifuged at 1800 rpm for 5 min to obtain pellet-containing cells. The supernatant was discarded and the pellet was re-suspended in 5 ml of Dulbecco's modified Eagle's medium. Single-cell suspensions of dental pulp were cultured in 24-well microtiter plate with Dulbecco's modified Eagle's medium and then incubated at 37°C in 5% CO2. The culture medium was changed for every 3 days until the cell confluence was achieved. Pulp tissue was minced into 1–2 mm fragments, and each piece was placed in 24-well microtiter plate with Dulbecco's Modified Eagle's Medium and then incubated at 37°C in 5% CO2. It should be considered that the total volume of the Dulbecco's Modified Eagle's Medium for outgrowth of cell must support the attachment of pulp pieces for further cell outgrowth (2–3 ml/well). Medium was changed after outgrowth was observed. The outgrown cells at confluence were sub-cultured at a ratio of 1:4. Cells were used for cytotoxic tests after the fourth passage.

Cytotoxicity assay

Ninety-five percent of curcumin (Hi-Media Laboratories, Mumbai, India) [Figure 1] was diluted with dimethylsulfoxide (Hi-Media Laboratories, Mumbai, India) in 100, 50, and 25 weight/volume %. Each concentration of curcumin was added in triplicate into 96-well microtiter plate containing the fibroblast culture. Cells without treatment served as a control group.
Figure 1: Crystalline curcumin (Hi-Media Laboratories, Mumbai, India)

Click here to view

The number of viable cells after 48 h incubation at 37°C in a humidified atmosphere of 5% CO2 and 95% air was determined by the methyl-thiazol-diphenyl-tetrazolium (MTT) assay. For this, 20 µL of 5 mg/ml MTT was added to all the 12 wells (10 mg in 1000 µL, 5 mg in 1000 µL, 2.5 mg in 1000 µL, and control, in triplicate) in 96-well microtiter plate and incubated at 37°C, 5% CO2 with 98% humidity for 4 h. At the end of the incubation period, the medium with MTT was removed, and 100 μL of dimethylsulphoxide was added to each well. The plate was shaken on the microplate shaker to dissolve the purple MTT-formazan. The relative viability of dental pulp fibroblasts was expressed with color intensity of the number in the experimental wells relative to that of control. Absorbance was recorded at 492 nm on a microplate reader with background subtraction at 620 nm (Lisaplus, Aspen Diagnostics Pvt., Ltd., Mumbai, India). The percentage of viable cells was determined by using the following equation: cell viability % = (mean absorbance of experimental wells/mean absorbance of control wells) × 100%.

   Results Top

All the tests were done in triplicate. The mean absorbance (optical density [OD]) and percentage values of cell viability at various concentrations of curcumin to human primary dental pulp fibroblasts are shown in [Table 1].
Table 1: Mean survival of human primary dental pulp fibroblasts as measured by MTT assay after exposure to various dilutions of curcumin for 48 h

Click here to view

   Discussion Top

The biologic and toxicologic properties of biomaterials are important for their clinical usage.In vitro cytotoxic screening as a primary factor of biocompatibility is determined by cell culture. In addition, in vitro tests are simple, reproducible, cost-effective, relevant, and suitable for evaluating the basic biological properties of dental materials.[10] Data on cell viability have long been obtained from in vitro cytotoxicity assays, and the MTT cell survival assay is widely used for measuring the cytotoxic potential of a compound.[11] The present study investigated the in vitro cytotoxicity of curcumin against primary dental pulp fibroblasts by MTT assay. No cytotoxicity was detected for curcumin at any of the concentrations used (25%, 50%, and 100%). The results revealed that the viability of primary dental pulp fibroblasts increased with an increasing concentration of curcumin [Figure 2]. It was observed that curcumin increased the cell proliferation of primary dental pulp fibroblasts, resulting in almost three times more cell viability than control in 50% and 100% groups.
Figure 2: Viability of primary dental pulp fibroblast at 25%, 50%, and 100% concentrations of curcumin

Click here to view

The choice of cell line for in vitro cytotoxicity screening assays remains debatable, as the apparent cytotoxicity of a material can be significantly affected by the cell line selected for the test. Nevertheless, primary cell cultures derived from the tissues of ultimate concern may arguably be more relevant to the clinical situation.[10]

The removal of the coronal portion of the pulp is an accepted procedure for treating both primary and permanent teeth with carious pulp exposures. The justification for this procedure is that the coronal pulp tissue, which is adjacent to the carious exposure, usually contains microorganisms and shows evidence of inflammation and degenerative change. The abnormal tissue can be removed, and the healing can be allowed to take place at the entrance of the pulp canal in an area of essentially normal pulp.[5] Placement of a medicament directly over the exposed pulp tissue has been suggested as a way to promote pulp healing and generate reparative dentin. If successful, this procedure precludes the need for more invasive, more extensive, and more expensive treatment.[12]

A therapeutic agent selected for the treatment of wounds should ideally improve one or more phases of healing without producing deleterious side effects.[7] Curcumin has been shown to have significant wound healing properties. Because of its low water-solubility, curcumin makes a suitable candidate for topical applications.[4] The wound healing potential of curcumin is attributed to its biochemical effects such as its anti-inflammatory,[13] anti-infectious,[14],[15] and antioxidant [16],[17] activities. The effects of curcumin topical treatment on different stages of cutaneous wound healing can be summarized as follows: (1) Stage of inflammation - inflammation is the crucial phase of the wound healing process. Uncontrolled inflammatory responses may lead to undesirable effects and subsequently tissue damage. Considering that tissue injury causes almost an immediate onset of acute inflammation, controlling inflammation is, therefore, desirable and can optimize the wound repair process. Curcumin has been shown to inhibit the production of tumor necrosis factor alpha and interleukin-1, two main cytokines released from monocytes and macrophages that play important roles in the regulation of inflammatory responses. Of equal importance is curcumin's ability to inhibit the activity of nuclear factor kappa-light-chain-enhancer of activated B-cells, a transcription factor that regulates many genes implicated in the initiation of inflammatory responses. (2) Stage of proliferation - the infiltration of fibroblasts into wound site is essential for granulation tissue formation/tissue remodeling, collagen production, and deposition of curcumin enhances fibroblast migration, granulation tissue formation, collagen deposition, and in general, re-epithelialization. Being apoptotic in the early phase of wound healing, curcumin eliminates unwanted inflammatory cells from the wound site. (3) Stage of remodeling - the re-organization and remodeling of the extracellular matrix are a prerequisite for wounds to heal completely. The extracellular matrix provides support to cells and it is composed of various proteins and polysaccharides including granulation tissue and collagen. Transforming growth factor beta (TGF-β) is an important cytokine that is involved in the repair, chemotaxis, and deposition of collagen in a wound site. Curcumin improves wound contraction by increasing the production of TGF-β and therefore increasing fibroblast proliferation.[7]

In this study, the cell viability values were consistently above 100% for all the test concentrations of curcumin. Apart from the action of curcumin, this could also be because the color of curcumin will add on to the OD values obtained.

The action of curcumin has never been reported for therapeutic uses in pediatric endodontic therapies. Possession of desired biological actions, pharmacological safety, and negligible cost makes curcumin an attractive agent to explore further for its potential uses in vital pulp therapy procedures in primary and permanent teeth and also for regenerative endodontic procedures in permanent teeth.

   Conclusion Top

Curcumin promotes cell viability and induces proliferation of primary dental pulp fibroblasts and has the potential to be developed into an economical and reliable medicament for vital pulp therapy, and it needs to be evaluated further in animal models for usage tests.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

   References Top

American Academy of Pediatric Dentistry. Clinical guidelines on pulp therapy for primary and young permanent teeth: Reference manual 2006-07. Pediatr Dent 2006;28:144-8.  Back to cited text no. 1
Fuks AB. Vital pulp therapy with new materials for primary teeth: New directions and treatment perspectives. J Endod 2008;34 7 Suppl: S18-24.  Back to cited text no. 2
Kulac M, Aktas C, Tulubas F, Uygur R, Kanter M, Erboga M, et al. The effects of topical treatment with curcumin on burn wound healing in rats. J Mol Histol 2013;44:83-90.  Back to cited text no. 3
Mohanty C, Das M, Sahoo SK. Sustained wound healing activity of curcumin loaded oleic acid based polymeric bandage in a rat model. Mol Pharm 2012;9:2801-11.  Back to cited text no. 4
McDonald RE, Avery DR, Jeffrey AD. Treatment of deep caries, vital pulp exposure, and pulpless teeth. In: McDonald RE, Avery DR, editors, Dentistry for the Child and Adolescent. 9th ed. Missouri, UK: Mosby, Inc. (an affiliate of Elsevier Inc.); 2011. p. 350.  Back to cited text no. 5
Chattopadhyay I, Biswas K, Bandyopadhyay U, Banerjee RK. Turmeric and curcumin: Biological actions and medicinal applications. Curr Sci 2004;87:44-53.  Back to cited text no. 6
Akbik D, Ghadiri M, Chrzanowski W, Rohanizadeh R. Curcumin as a wound healing agent. Life Sci 2014;116:1-7.  Back to cited text no. 7
Murray PE, García Godoy C, García Godoy F. How is the biocompatibilty of dental biomaterials evaluated? Med Oral Patol Oral Cir Bucal 2007;12:E258-66.  Back to cited text no. 8
van Wyk CW, Olivier A, Maritz JS. Cultured pulp fibroblasts: Are they suitable for in vitro cytotoxicity testing? J Oral Pathol Med 2001;30:168-77.  Back to cited text no. 9
Odabas ME, Ertürk M, Çinar Ç, Tüzüner T, Tulunoglu Ö. Cytotoxicity of a new hemostatic agent on human pulp fibroblasts in vitro. Med Oral Patol Oral Cir Bucal 2011;16:e584-7.  Back to cited text no. 10
Sumantran VN. Cellular chemosensitivity assays: An overview. Methods Mol Biol 2011;731:219-36.  Back to cited text no. 11
Hilton TJ. Keys to clinical success with pulp capping: A review of the literature. Oper Dent 2009;34:615-25.  Back to cited text no. 12
Liang G, Yang S, Zhou H, Shao L, Huang K, Xiao J, et al. Synthesis, crystal structure and anti-inflammatory properties of curcumin analogues. Eur J Med Chem 2009;44:915-9.  Back to cited text no. 13
Mun SH, Joung DK, Kim YS, Kang OH, Kim SB, Seo YS, et al. Synergistic antibacterial effect of curcumin against methicillin-resistant Staphylococcus aureus. Phytomedicine 2013;20:714-8.  Back to cited text no. 14
Singh RK, Rai D, Yadav D, Bhargava A, Balzarini J, De Clercq E. Synthesis, antibacterial and antiviral properties of curcumin bioconjugates bearing dipeptide, fatty acids and folic acid. Eur J Med Chem 2010;45:1078-86.  Back to cited text no. 15
Ak T, Gülçin I. Antioxidant and radical scavenging properties of curcumin. Chem Biol Interact 2008;174:27-37.  Back to cited text no. 16
Meng B, Li J, Cao H. Antioxidant and antiinflammatory activities of curcumin on diabetes mellitus and its complications. Curr Pharm Des 2013;19:2101-13.  Back to cited text no. 17


  [Figure 1], [Figure 2]

  [Table 1]

This article has been cited by
1 Fibroblast-targeting polymeric nanovehicles to enhance topical wound healing through promotion of PAR-2 receptor-mediated endocytosis
Yousong Lee, Seulgi Kim, Jihye Seo, Hyo Keun Kim, Yeong Pin Han, Eun Ju Park, Jin Oh Park, Chul-Su Yang, Jin Woong Kim
Biomaterials Science. 2023;
[Pubmed] | [DOI]
2 Gelatin–Curcumin Nanocomposites as a Coating for Implant Healing Abutment: In Vitro Stability Investigation
Solmaz Maleki Dizaj, Ali Torab, Shadi Kouhkani, Simin Sharifi, Ramin Negahdari, Sepideh Bohlouli, Shirin Fattahi, Sara Salatin
Clinics and Practice. 2023; 13(1): 88
[Pubmed] | [DOI]
3 MicroRNA and their implications in dental pulp inflammation: current trends and future perspectives
Manahil Maqbool, Nazmul Huda Syed, Giampiero Rossi-Fedele, Ismail Shatriah, Tahir Yusuf Noorani
Odontology. 2022;
[Pubmed] | [DOI]
4 Expression of immunomodulatory and tissue regenerative biomarkers in human dental pulp derived-mesenchymal stem cells treated with curcumin: an in vitro study
Malaksima Ayadilord, Mansoore Saharkhiz, Mohsen Naseri, Fariba Emadian Razavi
Molecular Biology Reports. 2022;
[Pubmed] | [DOI]
5 Dental pulp fibroblast: A star Cell
José Luis Álvarez-Vásquez, Cristina Paola Castañeda-Alvarado
Journal of Endodontics. 2022;
[Pubmed] | [DOI]
6 Impact of curcumin loading on the physicochemical, mechanical and antimicrobial properties of a methacrylate-based experimental dental resin
Patricia Comeau, Beatriz Panariello, Simone Duarte, Adriana Manso
Scientific Reports. 2022; 12(1)
[Pubmed] | [DOI]
7 Evaluating the Effect of Tideglusib-Loaded Bioactive Glass Nanoparticles as a Potential Dentine Regenerative Material
Akhil C. Rao, K. Vijay Venkatesh, Vidyashree Nandini, Dhanasekaran Sihivahanan, Ahmed Alamoudi, Hammam Ahmed Bahammam, Sarah Ahmed Bahammam, Bassam Zidane, Maha A. Bahammam, Hitesh Chohan, Nassreen H. Albar, Pradeep Kumar Yadalam, Shankargouda Patil
Materials. 2022; 15(13): 4567
[Pubmed] | [DOI]
8 The Potential Application of Natural Photosensitizers Used in Antimicrobial Photodynamic Therapy against Oral Infections
Shima Afrasiabi, Alireza Partoazar, Nasim Chiniforush, Ramin Goudarzi
Pharmaceuticals. 2022; 15(6): 767
[Pubmed] | [DOI]
9 Clinical and Radiographic Evaluation of Curcumin as an Obturation Material in Deciduous Teeth: A Randomized Controlled Trial
Shankar Paulindraraj, Hemalatha Ramkumar, Karthika Shankar, Charishma Sai Bommareddy, Senthil Dakshinamurthy, Trophimus Gnanabagyan Jayakaran
International Journal of Clinical Pediatric Dentistry. 2022; 15(S1): S35
[Pubmed] | [DOI]
10 A Systematic Review of Curcumin and its Derivatives as Valuable Sources of Antileishmanial Agents
Aishah E. Albalawi, Abdullah D. Alanazi, Iraj Sharifi, Fatemeh Ezzatkhah
Acta Parasitologica. 2021; 66(3): 797
[Pubmed] | [DOI]
11 Assessment and Characterization of Some New Photosensitizers for Antimicrobial Photodynamic Therapy (aPDT)
Laura Monica Dascalu (Rusu), Marioara Moldovan, Doina Prodan, Irina Ciotlaus, Violeta Popescu, Ioana Baldea, Rahela Carpa, Sorina Sava, Radu Chifor, Mindra Eugenia Badea
Materials. 2020; 13(13): 3012
[Pubmed] | [DOI]
12 A comparative study of the E. faecalis antibiofilm efficacy of photoactivated curcumin, chlorophyll and riboflavin
Riski Setyo Avianti, Sri Kunarti, Ari Subiyanto
Dental Journal (Majalah Kedokteran Gigi). 2020; 53(2): 62
[Pubmed] | [DOI]

Antiviral Activity of Chitosan Nanoparticles Encapsulating Curcumin Against Hepatitis C Virus Genotype 4a in Human Hepatoma Cell Lines

Samah A Loutfy, Mostafa H Elberry, Khaled Yehia Farroh, Hossam Taha Mohamed, Aya A Mohamed, ElChaimaa B Mohamed, Ahmed Hassan Ibrahim Faraag, Shaker A Mousa
International Journal of Nanomedicine. 2020; Volume 15: 2699
[Pubmed] | [DOI]


Print this article  Email this article


    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Article in PDF (604 KB)
    Citation Manager
    Access Statistics
    Reader Comments
    Email Alert *
    Add to My List *
* Registration required (free)  

    Materials and Me...
    Article Figures
    Article Tables

 Article Access Statistics
    PDF Downloaded518    
    Comments [Add]    
    Cited by others 13    

Recommend this journal

Contact us | Sitemap | Advertise | What's New | Copyright and Disclaimer | Privacy Notice
 © 2005 - Journal of Indian Society of Pedodontics and Preventive Dentistry | Published by Wolters Kluwer - Medknow 
Online since 1st May '05