INTRODUCTION

Recently conservative treatment of defective carious teeth is a mandatory.

Adhesive restorations facilitate the chemical bonding with tooth structure and
minimum reduction. Glass ionomer is one of unique adhesive and aesthetic
restoration. .

Glass ionomer are being used widely because of its excellent properties such
as chemical bonding to the tooth structure, biocompatibility and fluoride release.
On the other hand, there are a number of shortcomings for this group of materials
such as poor aesthetic, prolonged setting reaction, compromised mechanical
properties and weaker bond strength. Marginal deficiencies, wear, and secondary
caries are other considerations that jeopardize the long term performance of GIC
restorations in teeth(1).

In order to improve the properties and to overcome these shortcomings, active

research is in progress, such as in the addition of cellulose fibres, Hydroxyapatite
and fluoroapatite and nanotechnologies .(2) More recently, nanotechnologies have
been applied to the resin modified glass ionomer in the form of nanoparticles
(nanomers) and nanoclusters in fluoroaluminosilicate.

It was reported in 2007 that the flexural strength of a commercial GIC was
significantly improved by the addition of chitosan. Moreover, in the presence of
chitosan, it was found that release of fluoride ions from GIC was catalyzed.
Researchers have demonstrated its great potential for a wide range of uses due to its
versatile chemical and physical properties like biodegradability, biocompatibility,
antimicrobial activity, no toxicity. (3, 4)

1
 Hence, adding chitosan to glass ionomer adhesives might be improving the
physical and mechanical properties. Adding to that, it might enhance the antimicrobial
effect of glass ionomer adhesives.

2
 REVIEW OF LITERATURE

Saito et al 1999; reported that Glass ionomer cements (GIC) are unique
restorative materials with many uses in clinical practice. GIC are classified
according to their chemical formulation into two categories: conventional
( traditional) and resin-modified.(5)

Burgess, 2008; reported that Conventional glass ionomer cements undergo a

chemical self-setting acid-base reaction created by mixing an ion-leachable
(fluoride ion) fluoroaluminosilicate glass (powder) with an aqueous polyacrylic
acid or polycarboxylate acid (liquid). This advancement combined the advantages
seen with early silicate cements (chemically leachable (fluoride ion)
fluoroaluminosilicate glass [powder] and phosphoric acid [liquid]) and the
adhesive properties of zinc polycarboxylate cements (zinc-oxide [powder] and
polyacrylic acid [liquid]). This first type of GIC was developed by Wilson and
Kent(3). These changes and developing a higher-viscosity, thicker mix with the
chemistry of conventional GIC resulted in improvements in tensile strength,
compressive strength and fracture toughness; greater wear resistance and higher
fluoride release were also achieved .(6)

Conventional GIC used as tooth-colored restorative materials also have poorer

esthetics compared to composite resin. In order to expand the clinical uses of GIC,
resin was added to the formulation.

Croll and Berg, 2010; reported that resin-modified glass ionomer cement
(RMGI) chemistry was enhanced with the addition of water-soluble
photopolymerizable resin monomers, 2-hydroxyethylmethacrylate (HEMA) to the

3
acidic cement liquid, and for powder-liquid RMGI some manufacturers use
proprietary resin formulations .(7)

Mitra, 1991 and Uno et al 1996 ; The change in formulation of RMGI

allowed them to be dual-cured: self-setting and light-cured. When compared to
conventional GIC, resin modified glass ionomers provide for improved
physicomechanical properties, resistance to early contamination by moisture, less
microleakage, and improved adhesion to enamel and dentin combined with
significant improvement in esthetic properties.(8)

Leinfelder, 1993; One recent modified formulation includes more resin as

well as nanoparticles (Ketac Nano, 3M ESPE). In an effort to improve physical
properties for GIC as a posterior restorative, manufacturers also developed metal-
reinforced glass ionomers by adding silver amalgam alloy powder to GIC.(9)

The potential of remineralization of carious lesions in dentin in vitro by glass-

ionomer cements (GIC) has been related to the release of fluoride ions from the
restorative material. Inhibition of bacterial growth related to the amounts of fluoride
ion released. Reduction in bacterial adherence could be due to the release of fluoride
or silver or to an initially low material pH.(10)

Palenik, et al, 1992; studied in vitro the effects of various glass-ionomer

restorative materials and cements on the growth and plaque-forming abilities of oral
bacteria believed to be responsible for recurrent caries in humans. They found that
the glass ionomer had an inhibitory effect on growth and on adherence of oral
bacteria thought to be often involved with recurrent dental caries. (10)

4
 Chitosan a natural linear bio-polyaminosaccaride is a hydrolyzed
(deacetylated) derivative of chitin. Chitosan possesses reactive amino group,
reactive primary and secondary hydroxyl groups at C2, C3 and C6 positions
(11)
respectively. It has rigid crystalline structure through inter and intra molecular
hydrogen bonding. Chitosan is a weak base, insoluble in water and organic solvents,
but soluble in dilute aqueous acidic solution (pH < 6.5) .(12)

Petri et al 2007; reported that Chitosan carries a positive charge due to

easy availability of the free amino group and so reacts with surfaces/polymers with
negative charge, providing possibilities for covalent and ionic modifications,
thereby allowing improvement of properties. Due to its high biocompatibility and
hydrophilicity, Chitosan has been added to GIC.(13)

Petri, et al, 2006; studied the flexural strength and release of fluoride of
added chitosan to glass ionomer restoration ions , they found that tiny amount of
Chitosan improve flexural strength and more over release of fluoride Ions from
glass ionomer catalyzed.( 14)

Elsaka and Elnaghy, 2012; concluded that the modified self-etching primer
incorporating chitosan is a promising antibacterial primer which does not adversely
affect the bond strength of the Real Seal system to radicular dentin. (15)

Zhao, et al 2011; recommended addition of nanoparticles and nanofibers

chitosan, due to its significant biological and chemical properties such as
biodegradability, biocompatibility, bioactivity, and polycationicity. (16)

Diolosà, et al 2013; reported that Adding Chitosan to the experimental primer

enhanced the bond strength and interfacial nanoleakage . (17)

5
 Abraham, et al 2014; compared microleakage of glass ionomer cement
(GIC) and chitosan modified glass ionomer cement .The microleakage has been
decreased with Chitosan modified GIC. (18)

Rejane et al, 2009; were found that the antimicrobial Activity of Chitosan
has been dependant on polymeric molecular weight (MW) and degree of acetylation
(DA). They concluded that the lower the MW and the DA, the higher will be the
effectiveness on reducing microorganism growth and multiplication. (19)

Hence, the present study is a trial to add Chitosan to conventional glass ionomer
adhesive restoration to enhance its antimicrobial effect without adverse effect on
bond strength.

6
 AIM OF THE STUDY

This study aimed at:

1. Evaluation of anti bacterial effect of add chitosan to glass ionomer cement.

2. Evaluation of mechanical properties include compressive stress, surface
hardness, shear bond strength test.

3. Evaluation of physical properties include working time, setting time, water

sorption, water solubility, ph, fluoride release antibacterial release test, and test
of bacteriology.

4. Optimizing the proper ratio to enhance the antimicrobial without adverse

effect on bond strength.

7
 MATERIALS & METHODS

Materials

1-Conventional Glass ionomer cement.

2-Additative chitosan highly MW, (2-amino-2deoxy-(1->4)-B –d glucopyranan , sol.

Acetic acid 1%).

:Methods

Different proportions (0.5/1, 1/1, and 1.5/1 by volume chitosan / GI liquid)

from conventional glass- ionomer and chitosan will be prepared. The samples will
be shaped in the form of discs and will be divided into groups as follow:

Grouping of samples:

Group I: Conventional glass ionomer cement

Group II: Conventional glass ionomer cement and Chitosan (ratio a)

Group III: Conventional glass ionomer cement and Chitosan (ratio b)

Group IV: Conventional glass ionomer cement and Chitosan (ratio c)

The different prepared discs will be chemically and physically and

morphologically characterized as follow:

8
I. chemical Characterization:

1- Analyzing the surface chemistry by using x-ray diffractometer.

2- The organic chemical will be identified by Attenuated total reflectance Fourier
transform infrared spectroscopy (ATR).
3- Chemical compound of newly formed cement will be detected by using Nuclear
magnetic resonance (NMR).

II- Morphology:

The morphology of the surfaces will be characterized by the use of

scanning electron microscope.

III- Mechanical properties:

1- Surface hardness:

The surface micro-hardness of different prepared samples will be measured by

using Vickers's Hardness tester (Shimadzu, micro-hardness tester HMV-2E (344-
04109-22), V~100-120,220-240, 50-60 Hz, 300 VA). The indenter will be applied
with load 9.8 N for 10s. Three indentation readings will be made across the center
of each specimen.

2- Compressive stress test:

Specimens will be prepared in cylindrical Teflon mould of 8 mm height and

4mm diameter. By using universal testing machine compressive strength will be
determined.

3. Shear bond strength assessment:

9
 A universal testing machine will be used to test the strength of the glass
ionomer tooth interfacial bond.

IV- Physical properties characterization:

1- Setting time measurements:

P/l will be mixed and placed in Teflon mould has diameter 20 mm and thickness
1.5 ± 0.3mm. Setting time measurement will be performed by using indenter of
mass 400 ±5gm. Indenter have flat end of diameter 1.0 ±0.01mm will be applied to
the mix surface.

2- Working time measurement:

Samples will be prepared as mentioned previously. Working time will be

measured by using indenter of mass 28gm and diameter 2mm. flat ended needle tip
is cylindrical for distance approximately 5mm and perpendicular to long axis of
needle

3 -pH- measurements.

pH- measurements will be determined by using pH-meter. Sample are

prepared and placed in well sealed glass container definite amount of distilled water
to cover specimen. The pH of specimen will be monitored at different intervals.

4- Water solubility and water sorption test.

The split- Teflon mould of 20mm in diameter and 1.5mm of thickness will be
used. Testing will be done according to ISO 4049.The difference in weight will be
calculated before and after immersion in distilled water at 37°C for 24 hours. The

10
 values of water sorption (Wso) and solubility (Wsl) were calculated using the
following equations according to (ISO 4049:2000):

m1-m2

Wso =

　　　　 m0－m2

　Wsl =

where m0 is the specimen mass before immersion(mg), m 1 is the specimen mass

after immersion, m2 is the specimen mass after desiccation (mg), and V is the
specimen volume before immersion (mm3).

5- Test of bacteriology:

The antibacterial activity of the fabricated samples will be assessed using

Gram-negative bacteria Escherichia coli (E. coli) and Gram-positive
staphylococcus aureus by the microplate method as described by Li, et al ,
2013.(20)

6- Antibacterial release test:

All the prepared samples will be immersed in saline in well sealed plastic vials;
each sample in a vial. Each sample will be immersed in 50 ml of saline and kept at
37± 1ºC in an incubator for five days. Then, the samples will be rinsed by deionized

11
water and will be dried at 37 ºC± 1ºC. Release rate of antibacterial agent (chitosan) at
different time intervals (6, 12, 24, 48, 96 hrs) will be monitored by using ultraviolet
spectroscopy method, according to methodology described by Lv, et al,2014. (21)

7-Monitoring Fluoride release:

The samples are will be immersed in a well sealed plastic vials and incubated;
as previously mentioned test (IV-6). Then Fluoride release will be monitored at
different time intervals 1hr, 6hr, 12hr, 24hr, and 48hr

V- Statistical analysis:

Data will be collected, tabulated and then statistically analyzed using an

appropriate test.

12
 VI- Factorial designation of the study:

sted XRD FTIR NMR SEM St Wt PH Ws M-hard Shear Compressiv Release Fl-release Bacterial No.of
gps bond e stress rate culture sample

Gp1 1 1 1 1 5 5 5 5 5 5 5 5x5 5x5 5x2 99

R2 1 1 1 1 5 5 5 5 5 5 5 5x5 5x5 5x2 99

Gp3 1 1 1 1 5 5 5 5 5 5 5 5x5 5x5 5x2 99

Gp4 1 1 1 1 5 5 5 5 5 5 5 5x5 5x5 5x2 99

otal 4 4 4 4 20 20 20 20 20 20 20 100 100 40 396

mber

13
 REFERENCES.

1. Hatrick, C.D.; Eakle, W.S.; Bird, W.F. Dental Materials: Clinical Applications
for Dental Assistants and Dental Hygienists; Saunders: St. Louis, MO, USA,
2003. 48. Anusavice, K.; Phillips, R. Phillips’ Science of Dental Materials;
Saunders: St. Louis, MO, USA; 2003.

2. Moshaverinia, A.; Ansari, S.; Movasaghi, Z.; Billington, R.W.; Darr, J.A.;
Rehman, I.U. Modification of conventional glass-ionomer cements with
vinylpyrrolidone containing polyacids, nano-hydroxy and fluoroapatite to improve
mechanical properties. Dent. Mater. 2008, 24, 1381–1390. 4. Ito, M., 1991. In vitro
properties of a chitosan-bonded hydroxyapatite bone-filling paste. Biomaterials.
12(1): 41-45.

3. Kean T (Case Western Reserve University, Orthopaedics Department, Cleveland,

Ohio 44106, USA. tom.kean@case. edu), Thanou M. Biodegradation,
biodistribution and toxicity of chitosan. Adv Drug Deliv Rev 2010 Jan 31;62(1):3-
11.

4. Limapornvanich A (Dental Unit, Yala Regional Hospital, Yala 95000, Thailand),

Jitpukdeehodintra S, Hengtrakool C, Kedjarune-Leggat U. Bovine serum albumin
release from novel chitosan-fluoroaluminosilicate glass ionomer cement: Stability

14
and cytotoxicity studies. J Dent 2009 Sep;37(9):686-690 A.

5- Saito S, Tosake S, Hirota K. Characteristics of glass-ionomer cements. in

Advances in Glass-Ionomer Cements. Eds. Davidson CL, Mjor I. Quintessence
Publishing. 1999; pp. 15-50

6- Burgess JO. Fluoride-releasing materials and their adhesive characteristics.

Compend Contin Educ Dent. 2008; 29:82-94.

7- Croll TP. Berg JH. Glass-ionomer cement systems. Inside Dent. 2010; 6(8):82-
84.

8- Uno S, Finger WJ, Fritz U. Long-term mechanical characteristics of resin-

modified glass ionomer restorative materials. Dent Mater. 1996; 12:64-69.

Mitra SB. Adhesion to dentin and physical properties of a light-cured glass-

ionomer liner/base. J Dent Res. 1991; 70:72-74.

9- Leinfelder KF. Glass ionomers: current clinical developments. J Am Dent Assoc.

1993; 124:62-64. 9-Ito, M., 1991. In vitro properties of a chitosan-bonded
hydroxyapatite bone-filling paste. Biomaterials. 12(1): 41-45.

10-Palenik, C. J., Behnen, M. J., Setcos, J. C., & Miller, C. H. (1992). Inhibition of
microbial adherence and growth by various glass ionomers in vitro, 16–20 .

11- In-Yong Kim, Seog-Jin Seo, Hyun-Seuk MoonMi-Kyong Yoo, In-Young

Park, Bom-Chol Kim andChong-Su Cho, 2008. Chitosan and its derivatives for
tissue engineering applications. Biotechnology; 26: 1-21.

12- Hejazi, R. and M. Amiji, 2003. Chitosan-based gastrointestinal delivery systems

Journal ofControlled Release, 89: 151-165

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13. Petri DF, Donega J, Benassi AM, Bocangel JA (Instituto de Química,
Universidade de São Paulo, São Paulo, SP, Brazil. dfsp@usp.br). Preliminary study
on chitosan modified glass ionomer restoratives. Dent Mater 2007 Aug;23(8):1004-
1010.

14-Petri, D. F. S., Doneg, J., Benassi, M.& Bocangel, J. A. J. S. (2006). Preliminary

study on chitosan modified glass ionomer restoratives ´ a , Andr ´, 3, 1004–
1010. http://doi.org/10.1016/j.dental.2006.06.038

15- Elsaka, S., & Elnaghy, A. (2012). Effect of addition of chitosan to self - etching
primer: antibacterial activity and push - out bond strength to radicular dentin.
http://doi.org/10.7555/JBR.26.20120042.

16- Zhao, L. M., Shi, L. E., Zhang, Z. L., Chen, J. M., Shi, D. D., Yang, J., & Tang,
Z. X. (2011). Preparation and application of chitosan nanoparticles and
nanofibers. Brazilian Journal of Chemical Engineering, 28(3), 353–362. 17-
Forsten, L. (1977). Fluoride release from a glass ionomer cement, 504(Forsten
1976), 503–504.

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 17-Diolosà, M., Donati, I., Paoletti, S., Lenarda, R. Di, Breschi, L., & Cadenaro,
M. (n.d.). Bond stability of a chitosan-containing experimental adhesive.
Dental Materials, 29, e92. http://doi.org/10.1016/j.dental.2013.08.189.

18- Perchyonok, V. T., Grobler, S., Zhang, S., Olivier, A., & Oberholzer, T. (2013).
Insights into chitosan hydrogels on dentine bond strength and cytotoxicity,
2013(March), 75–82. 19- Goy, R. C., Britto, D. De, & Assis, O. B. G. (2009). A
Review of the Antimicrobial Activity of Chitosan, 19, 241–247.

19 Sasanaluckit, P., Albustany, K. R., Doherty, P. J., & Williams, D. F. (1993).

Biocompatibility of glass ionomer cements.

20- Li P, Zhang X, Xu R, et al. Electrochemically deposited chitosan/Ag complex

coatings on biomedical NiTialloy for antibacterial
application.SurfCoatingsTechnol.2013;232:370375.doi:10.1016/j.surfcoat.2013.05.03
7

21- Lv, H., Chen, Z., Yang, X., Cen, L., Zhang, X., &Gao, P. (2014). ScienceDirect
loaded chitosan / alginate multilayer on titanium substrates to inhibit biofilm
formation. Journal of Dentistry, 1–9. doi:10.1016/j.jdent.2014.06.003

17
 ‫‪.‬‬

‫المواد المستخدمة‬

‫االسمنت الزجاجى التقليدى ‪1.‬‬

‫اضافات مختلفه التركيز من الشيتوزان(‪0.5‬مللى و‪1‬مللى و ‪1.5‬مللى عاليه الوزن الجزىء المذابه فى‪2.‬‬
‫‪%‬حمض الخليك بتركيز ‪1‬‬

‫‪18‬‬
 ‫الهدف من الدراسة‬

‫تقييم االسمنت الزجاجى المحتوى على الشيتوزان كمضاد للببكتريا ‪1 .‬‬

‫تقييم الخواص الميكانيكية وتشمل اإلجهاد ضاغطة‪ ،‬وصالبة السطح‪ ،‬و اختبار قوة االسمنت‪2.‬‬

‫تقييم الخصائص الفيزيائية تشمل وقت العمل ووقت اإلعداد وامتصاص المياه‪ ،‬وقابلية الذوبان في الماء‪3.،‬‬
‫األس الهيدروجيني‪ ،‬اطالق الفلوريد ومضاد البكتريا‬

‫تحسين النسب المعدله لتحسين مضادات الميكروبات فى االسيمنت الزجاجى دون تأثير سلبي على قوة ‪4‬‬
‫‪.‬السيمنت‬

‫‪19‬‬
 ‫المقدمة‬

‫اصبح العالج التحفظى اساس‪¥¥‬يا فى معالج‪¥¥‬ه التس‪¥¥‬وس والحش‪¥¥‬و االص‪¥¥‬ق برابط‪¥¥‬ه كيم‪¥¥‬ا يئ‪¥¥‬ه م‪¥¥‬ع االس‪¥¥‬نان يقل‪¥¥‬ل‬
‫التسوس االسمنت الزجاجى هي مادة الصقة فريدة من نوعها و استعادة جمالية‪ .‬ويج‪¥¥‬ري اس‪¥¥‬تخدام االس‪¥¥‬منت‬
‫الزجاجي على نطاق واسع بسبب خصائصه الممتازة مثل الترابط الكيميائي لبنية السنه ‪ ،‬توافق مع لون الس‪¥¥‬نه‬
‫وإطالق الفلوريد‪.‬‬

‫‪  ‬‬

‫من ناحية أخرى هناك عدد من أوجه القصور لهذه المجموعة من الم‪¥¥‬واد مث‪¥¥‬ل س‪¥¥‬وء الجمالي‪¥¥‬ة لف‪¥¥‬ترة طويل‪¥¥‬ة ‪.‬‬
‫والذوبان الذى يضعف قوة االسمنت ‪ .‬موخرا تم اضافه الشيتوزان الذى يتميز بانه له ت اثير مض اد للبكتري ا‬
‫الى االسمنت الزجاجى لزياده مقاومه البكتريا ودراسه تاثيره على قوه رابطه االس منت م ع الس نه ومع دل‬
‫اطالق الفلوريد‪.‬‬

‫ومن ثم إضافة الشيتوزان السمنت الزجاجى قد يكون له تاثير على تحسين الخصائص الفيزيائية والميكانيكية‪.‬‬
‫إضافة إلى ذلك‪ ،‬قد تعزز تأثير االسمنت الزجاجى كمضاد للبكتريا ‪.‬‬

‫‪20‬‬
‫الالصق الزجاجى تحت التجربة المحتوى على الشيتوزان‬
‫‪.‬تقييم االختبارات الميكانيكية والفيزيائية والمضادة للبكتيريا ‪:‬‬

‫خطة بحث‬

‫مقدمــــــة إلي كلية طب األسنان‬

‫جامعة المنيا‬

‫لتسجيل درجة الماجستير‬

‫في خواص المواد الحيوية لألسنان‬

‫مقدمة من‬

‫الطبيبة‪ /‬رحاب صالح الدين سيد عبد اللطيف‬

‫بكالوريوس ‪2012‬‬
‫كلية طب األسنان‬

‫‪21‬‬
 ‫جامعه النهضة‬

‫قسم خواص المواد الحيوية‬

‫كلية طب األسنان‬

‫جامعة النهضة‬

‫‪2015‬‬

‫المشرفون‬

‫أ‪.‬م‪.‬د‪ .‬ياسر فتحي حسين‬

‫أستاذ مساعد خواص المواد الحيوية لألسنان‬

‫رئيس قسم خواص المواد الحيوية‬

‫كلية طب األسنان‬

‫جامعة المنيا‬

‫د‪.‬هبه عبد الحميد شلبى‬

‫مدرس خواص المواد الحيوية لألسنان‬

‫كلية طب األسنان‬

‫‪22‬‬
‫جامعة الفيوم‬

‫‪23‬‬