Nano-Mechanical Evaluation of Dental Hard Tissues Using Indentation Technique
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1 World Applied Sciences Journal 28 (10): , 2013 ISSN IDOSI Publications, 2013 DOI: /idosi.wasj Nano-Mechanical Evaluation of Dental Hard Tissues Using Indentation Technique 1 1 Muhammad Sohail Zafar and Naseer Ahmed 1 Dental Biomaterials, College of Dentistry, Taibah University, Madinah Al Munawwarah Saudi Arabia 2 Department of Mechanical Engineering, College of Engineering, Taibah University, Madinah Al Munawwarah Saudi Arabia Absract: Nanoindentation is a unique approach at can be used to measure e absolute hardness and elastic modulus of dental hard tissue. Enamel is e hardest materials however it changes from one point to e oer. The objective of is study was to calculate e elastic modulus and hardness profile of dental hard tissues from various anatomical locations using nano-indentation technique. First permanent mandibular premolars extracted for orodontic reason were used. Selected tee were dissected transversely into disc shape sections using a 2.3 mm standard grit cutting diamond disc (ISO806104). Prepared samples were stored in deionised distilled water and tested using Hysitron [TI 725 Ubi] nanomechanical testing instrument. Data collected was used to calculate e absolute hardness and elastic modulus. Key words: Nanoindentation Predentin Hardness Elastic modulus Enamel INTRODUCTION Dentin is an elastic, pale colored, slightly harder bonelike connective tissue at forms bulk of e too Tee are required for chewing and incising of food, and providing support to overlying brittle enamel [1, 7]. proper speech and aesetics [1]. The too is composed Dentin is composed of inorganics (70 wt. %) mainly of hard tissues (enamel, dentin and cementum) and act as hydroxyapatite [HA], 20 wt. % organic and 10 wt. % water a single mechanical unit to survive in a variety of stresses [1, 4, 5, 7, 8]. At dentino-enamel junction, ere is an and forces of mastication (up to ~ 700 N) [2]. Enamel is e extensive terminal branching of dentinal tubules hardest tissue of e body covering e crown dentin of producing many canaliculi and a highly interconnected e too [3-5]. This is a highly mineralized tissue wi 96 network system [1, 9]. Cementum is a bone like tissue at wt. % inorganic contents and 4 wt. % organic contents surrounds e root dentin of too and provides and water [4, 6]. The inorganic component is composed of attachment to e fibres of e periodontal ligament [2, 3, mainly calcium phosphate in e form of crystalline 7]. The ickness of e cementum layer is µm near hydroxyapatite [HA] and tiny amount of oer minerals e coronal dentin and increased up to 200 µm in e such as carbonate, magnesium, strontium, lead and apical area of root [1,7]. The biochemical composition of fluoride [7]. The presence of high inorganic content makes cementum is very similar to bone containing % enamel a very hard tissue to wistand e heavy forces inorganic in e form of hydroxyapatite and e rest is of mastication. However enamel is always supported by collagen and organic matrix [1,7]. Cementum can be dentin and becomes very brittle when unsupported [4, 7]. classified in mainly two types (cellular and acellular) Dentin is a unique dental tissue and is considered a vital depending on e presence or absence of cementum tissue even wiout e presence of a blood supply or forming cells [7]. innervations and has e ability to respond to stimuli like Enamel, dentin and cementum are classified as hard tactile, ermal or chemical changes [5]. materials however hardness significantly varies from each Corresponding Auor: Dr. Muhammad Sohail Zafar, Dental Biomaterials, College of Dentistry, Taibah University, Madinah Al Munawwarah Saudi Arabia. Tel:
2 oer and even hardness of one anatomical site may differ MATERIALS AND METHODS to oer site wiin e same tissue. For e purpose of repair, maintenance and replacement of ese natural Three permanent mandibular premolars extracted for tissues (in case of decay or disease), it is crucial to orodontic reasons were selected for is study. All tee understand e structural and mechanical properties in were examined carefully in order to make sure at roots relation to functional capabilities. Ignoring e properties are fully developed and ere is no caries, wear or any and parameters of natural too materials can result in oer paology. Samples having any visible crack were disaster or failure of man-made materials. The ideal also excluded. Tee were kept in 1 % hydrogen peroxide material to replace e lost enamel or dentin tissue will be solution for one day; followed by storage in deionised e one having matching physical and mechanical distilled water at 6 C for not more an one week. In order properties (such as hardness elastic modulus) to e too to explore mineralised dental tissues (DEJ, CDJ. structure. Predentin), all tee were dissected transversely into disc Hardness is simply defined as e resistance to shape sections using a 2.3 mm standard grit cutting permanent penetration. Hardness is an important property diamond disc (ISO806104). All samples were polished of dental materials at can influence on e processing using 1.0 and 0.5 micron diamond paste. Prepared sample such as cutting, finishing and polishing as well as were washed oroughly using running distilled water to functioning such as mastication. Conventional hardness remove any debris. Prepared samples were stored in testing is performed by applying a standardized force on deionised distilled water at 6 C and ph of an indenter to produce an indentation on e materials The sample to be tested was removed from e storage surface. The surface area, wid or dep of e medium and nanoindentation was performed at room indentation is calculated using a microscope to determine temperature. The testing of each sample was completed e hardness [10]. Nanohardness is calculated using e wiin 15 minutes considering prolonged testing may same principle however an indentation of submicron size cause moisture loss and affecting on e test results. is produced on e testing surface. A fixed quantity of force [usually 0.1mg to 5 g] is applied on e materials Nanoindentation Machine: Hysitron [TI 725 Ubi] using standard sized indenter. The indentation pa and nanomechanical testing instrument was used to perform dep is continuously monitored to calculate e e indentation (Figure 1). The indenter was equipped nanohardness and elastic modulus [10]. wi a 3-sided pyramidal Berkovich degree diamond In anisotropic composite materials such as too, probe. The system was placed on an anti-vibration table where we expect hardness to alter significantly from one to avoid any vibration during e test and any possible point to e adjacent; nanoindentation can be a very errors. For indentation, force (F) vs diaplacement (d) of useful tool. It can be helpful to study e hardness profile e load unload cycle was measured. The system was of too tissues (enamel, dentin, cementum and predentin) at submicron level at was not possible using conventional hardness testing meods. Furermore, e interface between ese tissues such as dentinoenamel junction (DEJ) and cementodentinal junction (CDJ) can be better studied to understand e functional transition from one tissue to e oer at submicron level. Similarly, cementum inness ranges form microns [11] and nanohardness will be a more reliable tool for measuring hardness profile for ese tissues. The objective of is study was to calculate e elastic modulus and hardness profile of dental hard tissues from various anatomical locations using nanoindentation technique. The data obtained can help clinicians to determine e materials of choice [at s having similar properties to e tissue to be replaced] hence preventing restoration failure due to mismatch of Fig. 1: Hysitron [TI 725 Ubi] nanomechanical testing mechanical properties. instrument 1394
3 Fig. 2: Indentaion curves produced using Hysitron [TI 725 Ubi] nanomechanical testing instrument; Ideal curve (A), Defective curve (B), Enamel (C) and Dentin (D) capable of calculating absolute hardness and reduced procedure. A load of 1N was used to carry out all e modulus (E). r The reduced modulus can be related wi indentations. The machine had an automated X-Y stage e modulus of elasticity of e specimen by e following wi a Z-Axis vertical motion control for e indenter tip. relation. A 10X optical camera is used to focus on e exact location of e indentation, after which e machine vsample 1 v = + indenter performs e indent. Examples of data curves are shown Er Esample Eindenter (Figure 2) to explain e data. For an ideal indentation, loading and unloading curve For a standard diamond indenter probe, E indenter is 1140 are continuous and smoo (Figure 2-A). However if e GPa and v indenter is Whereas e v sample is approximated indentation resulted in formation of crack, a step as 0.3 for e dental hard tissues [10]. Using ese values, formation will appear in e curve (Figure 2-B) leading to e E sample can be calculated false results. Such indentation results were excluded. Examples of enamel and dentin indention curves are The hardness has a nominal definition given by: represented (Figure 2-C and D). H = P max A RESULTS where P max is e maximum indentation force and A is e The mapping of hardness values have been shown resultant projected contact area at at load. The medium using color coding in e representative sections of e of indentation was air and longer exposures to air were mandibular premolar too crown and root (Figure 3). avoided to prevent e too from drying. The hardness of premolar too ranges from 0.01 to 7 GPa depending on e anatomical areas. The enamel at forms Testing Protocol: The prepared too sample was e outer shell of e crown has e highest value of mounted to e sample holder using cyanoacrylate hardness (5.70 to 7.18 GPa) at cusp tips, (4.20 to 5.39 GPa) adhesive. The sample holder was tightly screwed to e at occlussal surface and (4.02 to 5.03 GPa) on buccal and sample table so at ere is no rocking during e test lingual smoo surface (Figure 4). 1395
4 Fig. 3: Mapping of hardness zones rough various cross section of premolar too [Unit; GPa] Fig. 4: Nanoindentation of mineralized dental tissues and eir interfaces; Hardness (A) and Elastic modulus (B) The enamel hardness was furer reduced to (3.21 to tissues are significantly lower an enamel; ± GPa) in e cervical area and (2.23 to 2.89 GPa) near GPa, ±3.579 GPa and ±3,132 GPa for mantle, dentinoenamel junction (DEJ). The hardness of DEJ and primary and secondary dentin respectively. mantle dentin is very close as 1.24±0.15 GPa and 1.18±0.19 Cementum being softer material having elastic GPa respectively. Dentin at forms e bulk of e too modulus significantly lower an dentin and enamel is observed to be significantly softer an enamel. The (Figure 5). Coronal cementum elastic modulus is hardness of primary dentin is 0.819±0.101 GPa and 5.985± GPa, mid-root is 4.276± GPa, apical root secondary dentine is 0.691± GPa. Predentin is 3.468± Gpa and root tip is 2.965±0.285 GPa. represents organic matrix of dentin was observed to have The elastic moduli of interface tissues such as e lowest value of hardness (0.011±0.002 GPa). The dentinocementum junction and Predentin were found hardness of cementum varies roughout e root surface; lower (3.137±0.498 GPa and 1.394± GPa around too apex hardness remain 0.193± GPa, respectively). The elastic modulus of DEJ (32.569±5.569 apical ird remain 0.223± GPa and gradually GPa) was significantly lower an enamel however higher increases in e mid-root to (0.252± GPa) and an primary and secondary dentin. coronal ird of root (0.302± GPa). In terms of hardness, DEJ appears to be a transition The elastic modulus of dental tissues is directly zone (between enamel and dentin). The hardness of related to hardness and followed e similar pattern enamel was reduced near DEJ and furer reduces at DEJ (Figure 4). Enamel has e highest elastic modulus gradually to dentin (Figure 6). A different pattern was ( ± GPa) followed by occlusal enamel observed in case of cementum-dentin interface (CDJ). The (89.498± GPa) and bucco- lingual smoo surface hardness and elastic modulus of CDJ was recorded enamel (82.98± GPa). The modulus was furer 0.120±0.009 GPa and 3.139± GPa at are reduced for cervical enamel (77.673± GPa) and near significantly lower an e hardness and moduli of dentin DEJ (46.096±9.156 GPa). The elastic moduli of dentinal and enamel. 1396
5 Fig. 5: Nanoindentation of premolar root surface; Hardness (A) Elastic modulus (B) Fig. 6: Nanoindentation of dental tissue interfaces for hardness; Dentinoenamel junction (A), Cementodentinal junction (B) DISCUSSION inorganic contents of enamel are 96 % [11], making it e hardest material. The reason for e variation of mean This is e first study to describe e complete hardness values may be related to anisotropic nature of mapping of mandibular premolar too for hardness and enamel composites, fluctuation in mineral contents, elastic moduli using nanoindentation techniques. The orientation of HA crystal or enamel rods. These physical mean values of hardness and elastic moduli were properties make enamel a strong but brittle materials and significantly different among e ree tissues (enamel, is well supported by underlying softer and more elastic dentin and cementum), eir interfaces as well as at dentin [11]. Through e scalloped DEJ, heavy forces of various anatomical points wi in e same tissue. For mastication are transmitted across e dentin wiout example, enamel of e cusp tips and occlusal surface is mechanical failure. e hardest tissue, however a reduction of % in The variation of hardness and elastic moduli in hardness and elastic moduli was observed around e cementum is directly related to e structural differences. cervical areas of e premolars. Nature has adjusted e At root apex and apical part, cementum is mainly cellular hardness and elastic modulus as required for e having more cells, organic contents and fewer minerals applications. For example, hardness of enamel making it compared to e coronal part [11]. Coronal cementum has wear resistant and suitable to bear e forces of more mineral contents hence increasing e hardness and mastication. elastic modulus of e same tissue in is region. Hardness and elastic moduli are directly related to e Cementum is a diverse at exhibits continuous mineral contents in dental hard tissue [12, 13]. The remodeling roughout e life of a too [11], us 1397
6 mechanical properties may alter and should not Conflict of Interest: The auors declared no conflict of considered absolute. The cementum and dentin interface interest for conducting is research. in root is a distinct zone [ µm] at helps structural transition from harder dentin to cementum [14]. Similarly, REFERENCES CDJ facilitates e smoo transfer of masticatory stresses from periodontal ligament and cementum to e root 1. Ten Cate AR. Oral histology: Development, structure dentin wiout any mechanical failure. and function. 4 ed. St. Louis; London: Mosby, According to our search of scientific literature, ere 2. Jones, F.H., Tee and bones: applications of is no published research about e nano indentation of surface science to dental materials and related predentin. The ickness of predentin is microns biomaterials. Surface Science Reports, 42: at forms an interface between soft pulp tissue and 3. Fairpo, J.E.H., Heinemann dental dictionary. dentin [15]. The hardness and elastic modulus of 4 ed. Oxford: Butterwor-Heinemann. predentin has been reported here for e first time. Being 4. Restorative dental materials. 11 ed. St. Louis, Mo.; non-mineralized, predentin is an extremely soft material London: Mosby, [hardness; 0.011±0.002 GPa] at help in structural 5. Fundamentals of operative dentistry: A transition from soft connective tissues to hard connective nd contemporary approach. 2 ed. Chicago, Ill.; London: tissue. In addition, softer predentin also act as a cushion Quintessence Publishing, between hard dentin and delicate pulpal tissues. 6. nd Oral development and histology. 2 ed. New York: What is e hardness and elastic modulus of a Thieme Medical, premolar too? Interestingly, considering too as a 7. Nanci, A., Ten cate's oral histology: single unit, its hardness ranges from 0.01 to 7.18 GPa and Development, structure and function. 7 ed. St. elastic modulus ranges from 1.33 to GPa. This range Louis, Mo.; London: Mosby. might be widened furer by certain factors such as 8. Linde, A., Goldberg M. Dentinogenesis. Critical structural and individual variations, age, race and Reviews in Oral Biology & Medicine, 4: chemistry. These versatile mechanical properties of 9. United States. Public Heal Service. National enamel and dentin are due to significant anisotropic Institutes of Heal. National Institute of Dental nature of ese tissues [16]. Furer work is required to Research. An electron microscopic study of e early understand e biomimetic process involved and to stages of dentinogenesis. : pp. iv. 55. Washington, design new biomaterials to mimic e mechanical features similar to natural tissues. 10. Sakaguchi, R.L. and J.M. Powers, Craig's restorative dental materials. Philadelphia, PA: CONCLUSION Elsevier/Mosby. 11. Nanci, A., Ten cate's oral histology: Nanoindentation is a unique approach at can be Development, structure and function. 8 ed. St. used to measure e absolute hardness and elastic Louis, Mo.; London: Mosby. modulus of dental hard tissue. This is a reliable technique 12. Mahoney, E., A. Holt, M. Swain and N. Kilpatrick, to assess any alterations in tissue properties at e nano The hardness and modulus of elasticity of scale such as enamel-dentin interface or cementum-dentin primary molar tee:an ultra-micro-indentation study. interface. Enamel is e hardest materials; however J. Dent., 28: hardness reduces from cusp tip to cervical margin and 13. Malek, S., M.A. Darendeliler and M.V. Swain, from cusp tip to DEJ. Hardness and elastic modulus of Physical properties of root cementum: Part I. A new cementum gradually increases from apex towards meod for 3-dimensional evaluation. American cemento-enamel junction. Dentin did not show a lot of Journal of Orodontics and Dentofacial variation in hardness and elastic modulus except Oropedics, 120: predentin at has e lowest value. Biomaterials point of 14. Ho, S.P., M. Balooch, S.J. Marshall and G.W. view, ere is no artificial material at can be used for all Marshall, Local properties of a functionally mineralized tissues. The materials properties have to be graded interphase between cementum and dentin. modified to match wi e properties of e tissue to be Journal of Biomedical Materials Research Part A, replaced. 70A:
7 15. Garg, Nisha, S. Chandra and Garg, Amit, Bar-On, B. and H. Daniel Wagner, Enamel and Textbook of operative dentistry. New Delhi; London: dentin as multi-scale bio-composites. Journal of Jaypee Medical. e Mechanical Behavior of Biomedical Materials, 12:
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