Release characterization and biological effect of Glass Ionomer Functionalized with two different chlorohexidine derivatives: an in vitro study

HASSAN, Shaimaa M. Abdellah; KANDIL, Mohamed M.; El-REFAIE, Dina

doi:10.1590/1981-86372022001920200193

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ORIGINAL • Rev Gaúch. Odontol. 70 • 2022 • https://doi.org/10.1590/1981-86372022001920200193 copy

Release characterization and biological effect of Glass Ionomer Functionalized with two different chlorohexidine derivatives: an in vitro study

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Objective:

To evaluate the effect of adding two different chlorohexidine derivatives; chlorhexidine hexametaphosphate and chlorhexidine digluconate to glass ionomer at three different concentrations (0.25%, 0.75%and 1.5%) regarding the antibacterial effect, chlorhexidine release and fluoride release.

Methods:

A total of 405 specimens were prepared and tested after 7 days, 3 months and 6 months of storage in distilled water (n=5).For testing antibacterial effect, chlorhexidine release and fluoride release, the mix was packed in a ready-made Split Teflon molds to obtain disc-shaped specimen with dimensions 10 mm in diameter and 2 mm thickness according to ISO standardizations. One-way ANOVA and One-way repeated measure ANOVA test were used for statistical analysis of data.

Results:

The incorporation of chlorhexidine into ChemFil Superior glass ionomer cement in both derivatives has high significance ability to provide a long-term antimicrobial effect on Streptococcus mutans and Lactobacillius acidophilus. The chlorhexidine release was increased by adding chlorhexidine in both derivatives to GIC than the unmodified Glass-ionomer cement for study duration. However, the fluoride release was decreased in the modified specimens than the original one.

Conclusion:

Addition of chlorhexidine enhanced the antibacterial effect of the glass ionomer and chlorhexidine release. However, fluoride release was reduced than original

Indexing terms
Chlorhexidine; Chlorhexidine digluconate; Glass ionomer cement; Lactobacillius acidophilus; Streptococcus mutans

Brand name	Description	Composition	Manufacturer	Lot no.
Glass ionomer cement GIC	ChemFil^® Superior Powder/ liquid GIC	1 g powder contains: 1.(0.84 g) Aluminium-sodium-calcium-fluoro-phosphoro-silicate (18: 9: 8: 16: 3: 46) 0.84 g 2.(0.15 g) Polyacrylic acid (MW 30000-45000) 0.15 g Liquid: 10 ml demineralized water	DENTSPLY DeTrey GmbH, Konstonz, Germany	1709000428
Chlorhexidine digluconate 20% aqueous solution	Colorless or pale yellowish liquid. It is miscible with water, soluble in acetone and in alcohol. The structural formula: C22H30Cl2N10, 2C6H12O7	Aqueous sol of chlorhexidine which cannot be isolated as a solid, Soluble in water to at least 50%(W/V), with PH range of 5-8, density 1.06 to 1.07 Melting range between 132 °C TO 136 °C	BAJAJ Healthcare LTD, Gujarat, India	CS-00400216
Sodium hexametaphosphate HMP powder	White crystals, odorless, hexamer of composition (NaPO₃)₆. Sodium hexametaphosphate, mixture of polymeric metaphosphates, formula: (NaPO₃)₆	65-70% P2O5 basis Natrium hexametaphoshate, calgon, phosphate glass, water soluble, polyphosphate sodium salt, Soluble in water, Melting range 628 °C	Sigma-Aldrich Co., St. Louis, MO, USA)	BCBN0343v
Distilled water	Purified water USP35	50 ml purified water for oral use	FIPCO, Borg Elarab, Alexandria, Egypt	29050/2013

Type of material	Method of specimen’s preparation	Type of chlorhexidine	Method of chlorhexidine application
Conventional glass ionomer cement specimens	- Ratio of 1 scoop powder: 1 drop of liquid was mixed with P/L ratio (wt/wt) of 7.4: 1 - Half the powder was incorporated first into the liquid as quickly as possible (5seconds) and then the remainder was added and spatulated for 20 second to form a thick putty-like consistency. - Total working time was 2 minutes. - Total setting time was 2-3 minutes.	- no chlorhexidine added	- no application done.
Glass ionomer containing chlorhexidine digluconate specimens	Ratio of 1 scoop powder: 1 drop of liquid containing chlorhexidine digluconate was mixed with P/L ratio (wt/wt) of 7.4: 1 - Half the powder was incorporated first into the liquid as quickly as possible (5seconds) and then the remainder was added and spatulated for 20 second to form a thick putty-like consistency. - Total working time was 2 minutes. - Total setting time was 2-3 minutes.	- chlorhexidine digluconate liquid	- C hlorhexidine digluconate was added to the distilled water at concentrations 0.25 %, 0.75% and 1.5% of CHX digluconate.
- Glass ionomer containing chlorhexidine hexametaphosp-hates specimens	- Ratio of 1 scoop powder: 1 drop of liquid containing chlorhexidine hexametaphosphate was mixed with P/L ratio (wt/wt) of 7.4: 1 - Half the powder was incorporated first into the liquid as quickly as possible (5seconds) and then the remainder was added and spatulated for 20 second to form a thick putty-like consistency. - Total working time was 2 minutes. - Total setting time was 2-3 minutes.	- chlorhexidine hexametaphosphate powder	- chlorhexidine hexametaphosphate (HMP) was prepared by mixing aqueous 10 ml solutions of CHX digluconate and sodium HMP in a glass beaker and vigorous stirring for approximately 1 min, then the preparation was allowed to settle for 24 h to produce a precipitate. - The precipitate was filtered from the flask and discarded leaving a concentrated suspension. The suspension was centrifuged at 4760 g for 30 min, and then filtered again to discard the new precipitate. The remaining paste was removed from the centrifuge tubes using a spatula. The paste was added to distilled water to achieve concentrations of 0.25 %, 0.75% and 1.5% of CHX hexametaphospahte liquid.

Concentration of different chlorohexidine derivatives	Time of measurement (mean±SD)			p-value
Concentration of different chlorohexidine derivatives	7 days	3 months	6 months	p-value
p-value	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC)	24.00±5.48^Ba	4.40±6.02^Db	0.00±0.00^Db	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC HMP0.25%)	46.00±4.18^Aa	24.00±4.18^Bb	13.20±1.30^Cc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC HMP0.75%)	47.00±4.47^Aa	28.00±2.74^Bb	16.20±0.84^Bc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC HMP 1.5%)	54.00±5.48^Aa	38.00±2.74^Ab	17.60±0.55^Ac	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 0.25%)	28.00±2.74^Ba	6.60±6.02^Db	0.00±0.00^Db	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 0.75%)	46.00±4.18^Aa	14.60±0.55^Cb	0.00±0.00^Dc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 1.5%)	50.00±6.12^Aa	14.60±0.55^Cb	0.00±0.00^Dc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**

Concentration of different chlorohexidine derivatives	Time of measurement (mean±SD)			p-value
Concentration of different chlorohexidine derivatives	7 days	3 months	6 months	p-value
p-value	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC)	25.00±7.07^Da	4.40±6.02^Cb	0.00±0.00^Db	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC HMP 0.25%)	39.00±7.42^BCa	22.00±4.47^Bb	11.40±0.55^Cc	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC HMP 0.75%)	44.00±5.48^Aba	25.00±3.54^ABb	14.40±0.55^Bc	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC HMP 1.5%)	54.00±5.48^Aa	35.00±5.00^Ab	16.40±0.55^Ac	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC DI 0.25%)	28.00±2.74^CDa	6.60±6.02^Cb	0.00±0.00^Db	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC DI 0.75%)	46.00±4.18^Aba	7.00±6.40^Cb	0.00±0.00^Db	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC DI 1.5%)	52.00±7.58^Aa	7.00±6.40^Cb	0.00±0.00^Db	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).

Concentration of different chlorohexidine derivatives	Time of measurement (mean±SD)			p-value
Concentration of different chlorohexidine derivatives	7 days	3 months	6 months	p-value
p-value	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).	<0.001^* * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).
(GIC)	0.06±0.03^Da	0.01±0.00^Ea	0.01±0.00^Da	0.156ns
(GIC HMP 0.25%)	2.85±0.47^Ca	1.87±0.58^BCab	0.83±0.09^Cb	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC HMP 0.75%)	4.09±0.59^Ba	2.50±0.36^Bb	1.25±0.21^Bc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC HMP 1.5%)	5.91±0.34^Aa	3.41±0.43^Ab	1.84±0.41^Ac	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 0.25%)	3.14±0.36^Ca	0.89±0.08^Db	0.01±0.01^Dc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 0.75%)	4.07±0.64^Ba	1.90±0.61^BCb	0.01±0.01^Dc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**
(GIC DI 1.5%)	4.68±0.38^Ba	1.25±0.23^CDb	0.04±0.03^Dc	*<0.001^ * ; significant (p ≤ 0.05) ns; non-significant (p>0.05).**

Concentration of different chlorohexidine derivatives	Time of measurement (mean±SD)			p-value
Concentration of different chlorohexidine derivatives	7 days	3 months	6 months	p-value
p-value	<0.001^* * ; significant (p ≤ 0.05)	<0.001^* * ; significant (p ≤ 0.05)	<0.001^* * ; significant (p ≤ 0.05)
(GIC)	7.90±0.72^Aa	1.96±0.45^Ab	0.66±0.35^Ac	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC HMP 0.25%)	4.32±1.42^CDa	1.31±0.18^Bb	0.45±0.27^ABc	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC HMP 0.75%)	3.09±0.62^Da	1.06±0.05^Bb	0.18±0.11^Bc	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC HMP 1.5%)	4.11±0.56^CDa	1.08±0.07^Bb	0.17±0.05^Bc	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC DI 0.25%)	6.35±0.80^ABa	1.91±0.37^Ab	0.56±0.23^ABc	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC DI 0.75%)	4.93±1.11^BCa	1.22±0.12^Bb	0.44±0.08^ABc	*<0.001^ * ; significant (p ≤ 0.05)**
(GIC DI 1.5%)	4.20±0.76^CDa	1.10±0.09^Bb	0.18±0.13^Bc	*<0.001^ * ; significant (p ≤ 0.05)**

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[1] Correspondence to: SMA Hassan. Email: shaimaamortada13@gmail.com.