1014 D3000 new imprint Word template Vol 13, No 1 (2025) ISSN 2167-8677 (online) DOI 10.5195/d3000.2025.1014 http://dentistry3000.pitt.edu Effect of Strontium Titanate Addition on Antifungal, Physical and Me- chanical Properties of Soft-Liners Zakaria Alabed, Aseel Mohammed Al-Khafaji College of Den*stry, University of Baghdad, Baghdad, Iraq Abstract Objec8ve: So? denture liners are essenEal in prosthodonEcs due to their ability to cushion masEcatory forces and provide comfort to paEents with compromised oral Essues. Despite their clinical benefits, convenEonal so? liners suffer from significant limitaEons, including poor bond strength with the denture base, surface degradaEon, microbial colonizaEon, and limited mechanical resilience. The goal was to assess improvements in shear bond strength and other related surface properEes such as surface hardness, and a reducEon in C. albicans adherence. Material and Methods: StronEum Titanate (SrTiO₃) nanoparEcles were obtained from, and a self-cured so? denture liner was sourced from. All procedures were performed under condiEons simulaEng intraoral temperature, following the manufacturer’s instruc- Eons. A dry-heat oven was used for sample condiEoning. A total of 45 specimens were fabri- cated and equally allocated into three experimental groups containing 0%, 1%, and 1.5% con- centraEons of stronEum Etanate (SrTiO₃), respecEvely. Results: The results indicated that SrTiO₃ can play a valuable role in the development of advanced so? lining materials with enhanced durability, biocompaEbility, and clinical effecEveness. Conclusion: The incorporaEon of StronEum Titanate (SrTiO₃) nanoparEcles at concentraEons of 1% and 1.5% significantly improves the mechanical properEes, surface rough- ness, and reducEon in anEfungal behav- ior of acrylic-based so? denture liners. Open Access Cita%on: Alabed Z, et al. (2025) Effect of Stron%um Titan- ate Addi%on on An%fungal, Physical and Mechanical Pro- por%es of SoH-Liners. Den%stry 3000. 1:a001 doi:10.5195/d3000.2025.1014 Received: August 6, 2025 Accepted: August 20, 2025 Published: September 10, 2025 Copyright: ©2025 Alabed Z, et al. This is an open access ar%cle licensed under a Crea%ve Commons ATribu%on Work 4.0 United States License. Email: Zakariya.Azam2401m@codental.uobagh- dad.edu.iq Introduc)on Complete and partial removable dentures have used soft denture liner materials for over a century to disperse functional loads on the supporting tissues of the denture. This has allowed for material manipulation, improving the materials' physical, chemical, biological, and electrical qualities in compar- ison to their larger-scale counterparts [1]. Function, comfort, and appearance all affect how effective full or partial dentures are. A precise denture foundation that appropri- ately adapts to the underlying tissue is cru- cial. When building complete dentures, this is a crucial goal [2]. A complete denture is used when a person has lost all of their teeth, while a partial denture is used if they still have some teeth [3]. The fundamental rea- son removable dentures may retain their support, stability, and retention is because their bases are Fine-suited to the oral mucosa [4]. Strontium titanate (SrTiO₃) is a ceramic oxide with remarkable mechanical and ther- mal properties, making it valuable in various dental applications. Its incorporation en- hances strength, hardness, and durability of dental materials without affecting their es- sential Flexibility. This contributes to im- proved performance and longevity of resto- rations in the oral environment [5]. Progres- sive bone resorption compromises denture Fit, resulting in discomfort and even signiFi- cant pain, ultimately affecting the patient’s comfort and quality of life [6]. Material and Methods A total of 45 specimens of a self-cured, room- temperature polymerizing denture liner were prepared for evaluating Candida albi- cans adherence, surface roughness, and shear bond strength. The specimens were di- vided equally into three groups containing 0%, 1%, and 1.5% concentrations of stron- tium titanate (SrTiO₃), as shown in the table 1. Surface roughness was assessed using a proFilometer, while C. albicans adherence was quantiFied via optical density measure- ments using a spectrophotometer. Shear bond strength was evaluated using a testing machine (WDW-50). Effect of StronEum Titanate AddiEon on AnEfungal, Physical and Mechanical ProperEes of So?-Liners Vol 13, No 1 (2025) DOI 10.5195/d3000.2025.1014 http://dentistry3000.pitt.edu 2 Statistical analysis Statistical comparisons between groups were performed using one-way ANOVA in the SPSS software, with a signiFicance level set at α = 0.05. The analysis revealed statistically signiFicant differences across all tested parameters, in- dicating that the incorporation of SrTiO₃ had a substantial effect on the mechanical and bi- ological properties of liner material as shown in Table 1. Shear bond strength test Sample design Using acrylic block dimensions, the shear bonding strength between the soft lining ma- terial and the acrylic denture base was as- sessed. Two heat-cured acrylic blocks meas- uring 75*25*5 mm in diameter, length, width, and Thickness, as well as a stopper that was roughly 3 mm deep, made up the shear bond sample, as shown in Figure 1 [7]. To apply reline material, the two acrylic blocks are positioned one on top of the other, leaving a 25 mm*25 mm* 3 mm space be- tween them. The handle needs to be pre- cisely 13 mm thick to guarantee that the ap- plied forces are parallel and that the testing machine is gripping the sample well [8]. Preparation of the heat-cure acrylic sam- ples Mold preparation It is possible to create acrylic blocks using an aluminum template. It was manufactured by employing substances mold and a machine that cuts by laser, in addition to other meas- uring tools that it is essential to the Inferior portion includes both the pattern and the mold, it belongs to the dental flask of course. However, it includes a dental stone of the third type which originally had been de- signed to comply with the instructions of the producing party (p/L) ratio of 25ml/100q). As soon as the stone is set, both the stone and the Aluminum model would be covered with an isolating material and thus be liable to dry. The higher section of the flask would later be placed over the lower part. Next, a stone that was recently mixed would be added up. The aluminum sculpture was extracted from the mold. Consequently, we can open the flask when we had solidified the stone. Proportioning and mixing of heat cure acrylic MAARC DENTAL rapid heat-cure acrylic was used to create the acrylic sample. The mix- ture was combined in a dry, clean glass con- tainer with a powder-to-liquid monomer ra- tio of 10 cc of powder per 5 mL, as directed by the manufacturer. After being covered, the mixture was let to solidify into dough. Packing We covered the two parts of a flask with an isolating material that will be dried up later. Next, we erased the sculpture and locate the dough like acrylic resin into a hollow room. Later, we cover the acrylic with a polythene layer. We close thus the flask. A hydraulic pressure is employed to scatter the sub- stance equally within the mold. So, it can get pressure step by step. Curing We used a digital water bath, as shown by the producer, in order to cure the material of the acrylic resin. The resin is then placed in a water bath at 74°C for approximately 120 minutes. Subsequently, the temperature is increased to 100 °C to complete the curing process. Since cooling is achieved at normal heat of room. For 30 minutes, we soak the metal flask in a faucet water, then we can get it from the flask. Preparation of final samples for shear bond strength test Mold preparation One sample was created by using two acrylic blocks to test the shear bonding strength. When the two blocks are brought together, they make a space of 25mm × 25mm × 3mm (length, width, and depth accordingly), as shown in Figure 1. This space is then filled with material. Then, the samples were en- tirely covered with laboratory silicone and allowed to set properly the acrylic blocks and silicone mold are placed together after the silicon has been set in a custom-made flask filled with freshly mixed type III dental stone The custom flask was made from and consisted of upper and lower parts with a di- mension of (33cm length, 23cm width, and 4cm height) for each part. The parts came to- gether and tightly screwed until edge-to- edge contact was achieved. Incorporation of SRTIO3 In a dry, clean glass container, the strontium titanate nanoparticles were weighed with an electronic balance. After being measured us- ing a medical syringe, the soft-liner mono- mer was applied to the nanoparticles. After that, the mixture was put through a probe sonication device. For 3 minutes at 120W and 60 KHz to break up the nanoparticles into separate particles [9]. To avoid particle aggregation, the resultant SRTIO3-soft liner monomer dispersion was combined right away with soft liner powder. By deducting their weight from the soft-liner polymer powder, it was thought that adding SRTIO3s to the monomer would result in the proper P/L ratio. Soft liner application We weigh the (SrTiO₃) within a spotless, dry glass container using an electronic balance. Next, we measure employing a medical sy- ringe, thus later we expose the soft-liner monomer dispersion suddenly with soft liner powder. It was believed, through reduc- ing the weight from the soft-liner polymer powder, that if we add SrTio3 soft liner mon- omer would cause the cause the suitable P/L percentage, as shown in Table. Curing According to the manufacturer's instruc- tions, the material should be placed at a tem- perature like that of the mouth. The samples were placed in the designated places in the mold and transferred to the oven for 4 to 5 minutes at a temperature of 37 degrees Cel- sius. The samples were then removed from the oven and removed from the mold. Testing procedure The models underwent testing at, utilizing an Instron testing machine (WDW-50 from Laryee Technology Co., a computer-con- trolled electronic universal testing machine) with a load cell possessing a capacity of 50 KN and the crosshead speed set to 0.5 milli- meters per minute, as shown in Figure 1. Ac- cording to ASTM specification D-638m, the shear bond values were determined by di- viding the ultimate load needed to cause sample failure by the samples' cross-sec- tional area [10]. Surface Hardness Test Sample design and preparation The disc like plastic pattern was manufac- tured to comply with ISO 10139, 2, 2016, it had a diameter of 35 millimeter and 6 thick- ness. We employ the plastic models to create laboratory soft denture liner manipulation samples as templates. Those soft lining sub- stances of samples were created, mixed, stuffed in and dried in compliance with the manual [11]. Testing Surface hardness testing was carried out in accordance with ISO 10139-2:2016, which is the International Organisation for Standard- isation (ISO) guideline for permanent soft denture liners. Before being measured, all samples were incubated for five minutes in a perfect oven setting that about matched the manufacturer's recommended oral tempera- ture [12]. The hardness was measured using the Shore A durometer (produced by Zwick, Germany); it was calibrated in compliance with ASTM D2240. We reported the data us- ing shore units. The device is comprised of a 1.6 mm cylinder that gradually shrinks to a blunted indenter with a diameter of 0.8 mm. An indentation point-holding lever is con- nected to a scale with gradations from 0 to 100 units. Zero signifies that the indenter has completely pierced the sample, whereas 100 denotes no penetration at all. To test the discs, five locations were selected. The sam- ple was loaded evenly across its surface, away from its edges, to determine the aver- age shore hardness [13]. Fabrication of soft-liner specimens to evaluate C. albicans adherence ability Specimen preparation Effect of StronEum Titanate AddiEon on AnEfungal, Physical and Mechanical ProperEes of So?-Liners Vol 13, No 1 (2025) DOI 10.5195/d3000.2025.1014 http://dentistry3000.pitt.edu 3 Disk-shaped plastic patterns, each measur- ing 10 mm in diameter and 2 mm in thick- ness, were used to prepare the soft-liner specimens [14]. These plastic disks were po- sitioned in the lower half of a dental flask al- ready filled with dental stone. Once the stone had fully set, the surface—including the plas- tic patterns—was coated with a thin layer of separating medium and allowed to dry. Sub- sequently, the upper half of the flask was as- sembled and filled with dental stone using vibration to eliminate any air bubbles, then sealed with its cover. After the second layer of stone had completely set, the flask was opened, and the plastic disks were carefully removed [15]. Proportioning and mixing of room tem- perature Acrylic Based Soft liner Control negative specimens’ preparation According to the manufacturer's instructions (powder-to-liquid ratio of 2.2 g:1.8 ml), the required amounts of soft-liner powder and liquid, as shown in Figure 3 were measured and mixed in a clean, dry glass jar, then cov- ered with a lid. Both the upper and lower halves of the dental flask were coated with a separating medium to prevent the soft liner from adhering to the stone. Once the mixture reached the dough stage, it was hand- kneaded and adapted onto the lower half of the flask in preparation for packing. Positive Control Specimen preparation To comply with the producer’s recommen- dation, powder to liquid percentage of 8.6, 8.1ml.wt% of Nystatin powder was precisely calculated and taken out of the total weight of the sift liner powder. We add the left soft liner powder with Nystatin and mix all with the soft liquid by a tiny electric manual mixer for 60 seconds. By doing so, the targeted quantity of monomer will be added up to the mixture [16]. Incorporation of SRTIO3 nanoparticles We weigh the SrTiO3 nano particles within a spotless dry glass container then add up the soft-liner monomer. The total mixture will be exposed to probe sonification for almost 3 minutes at 120 and 60 Khz to scatter. Those nano particles by dividing up any agglomer- ates and attaining uniform distribution [10]. To prevent particle aggregation, the result- ing SrTiO₃–soft liner monomer suspension was immediately mixed with the soft-liner powder. To maintain the manufacturer's specified powder-to-liquid (P/L) ratio, the weight of the SrTiO₃ nanoparticles was sub- tracted from the total weight of the soft-liner powder. A medical syringe was used to measure the volume of the monomer, while an electronic balance was employed to accu- rately weigh the SrTiO₃ nanoparticles and soft-liner powder. Packing Since the soft liner gets into the dough pe- riod. We knead it by hand and used this for the ready mold. We put a polyethylene sheet over the substance. The lid will be placed on the top part of the flask. To guarantee the uniform scattering of the soft lining material in the mold to get rid if any extra value we use hydraulic press to maintain permanent pressure of 100kg/cm2. Fir about 5 minutes. The mixture will be positioned inside an oven with a temperature that’s close to the oral environment. This will be achieved to comply with the manual recommendations. When cutting is over, we opened the flask af- ter I was totally cold, and we took the speci- men out of the mold [10]. After curing com- pletion, the flask was opened when became completely cold and the specimens were re- moved from the mold. Finishing and Sterilization After trimming the excess material from the specimens using a sharp blade, they were finished using a 240-grit silicon polishing bur followed by fine-grit sandpaper. The specimens were then rinsed with distilled water, properly positioned, and sterilized under UV light for 20 minutes on each side at a wavelength of 254 nm. Finally, the speci- mens were stored in test tubes containing 250 ml of distilled water for 24 hours [10]. Isolation of C. albicans Candida albicans was collected from the oral cavities of patients showing clinical signs of denture stomatitis who visited the Prostho- dontics Clinic at the College of Dentistry, Kufa University, for treatment. Sterile cotton swabs were used to gently collect samples from oral lesions, which were then cultured on Sabouraud Dextrose Agar (SDA), a stand- ard medium for fungal growth. The culture plates were incubated at 37°C under aerobic conditions for 48 hours. Following incuba- tion, the fungal isolates were preserved at 4°C for future testing [17,18]. Preparation of Sabouraud dextrose agar According to the manufacturer’s instruc- tions, 62 g of Sabouraud Dextrose Agar (SDA) was weighed and fully dissolved in 1000 ml of distilled water. The mixture was then sterilized using an autoclave at 121°C and 15 psi for 15 minutes. After sterilization, the medium was allowed to cool to approxi- mately 47 °C to prevent damage to the Petri dishes. To inhibit bacterial growth, 0.05 g of the broad-spectrum antibiotic chloramphen- icol was added per 1000 ml of prepared me- dium. The SDA was then poured into Petri dishes, allowed to cool and solidify, and sub- sequently stored at 4 °C until use [19]. Identification of C. albicans Morphological examination A smooth, creamy, and pasty Candida colo- nies appear in Sabouraud dextrose agar me- dium. Microscopical examination A small amount from a single isolated Can- dida colony was picked and mixed with a drop of normal saline on a clean glass slide to create a suspension. This suspension was evenly spread across the slide, left to air dry at room temperature, and then heat-fixed by passing the slide several times through the flame of a Bunsen burner. Gram staining was performed according to the method outlined by Marler (2001) [23], as follows: 1. The slide was stained with crystal violet for one minute, then rinsed with distilled wa- ter. 2. Gram’s iodine was applied for one mi- nute and rinsed off. 3. Decolorization was carried out using ac- etone-alcohol until the dark violet color lightened, followed by a rinse with distilled water. 4. The slide was counterstained with safra- nin for one minute, rinsed again, and left to dry. Microscopic examination under a light microscope revealed Candida as round or oval cells. The prepared slide was examined under a light microscope, where Candida ap- peared as round or oval-shaped cells [26]. Germ tube formation A loopful of yeast cells was taken from a sin- gle colony and suspended in tubes contain- ing 0.5 ml of serum. The tubes were then in- cubated at 37 °C for 3 hours. After incuba- tion, a drop of the suspension was placed on a clean glass slide and examined under a light microscope to observe the presence of germ tubes [22]. Biochemical IdentiJication The VITEK 2 system is a fully automated de- vice that utilizes sensitive Fluorescence- based technology for the identiFication of mi- croorganisms. Before testing, a yeast suspen- sion was prepared in sterile saline, adjusted to a turbidity equivalent to a 2.0 McFarland standard, as veriFied using the DensiChek in- strument. The VITEK ID-YST card was then automatically Filled with the suspension, sealed, and incubated at 35.5°C for 18 hours within the VITEK 2 system. Optical density readings were taken automatically every 15 minutes throughout incubation. After com- pletion, the results were compared with the device’s database, enabling identiFication of the unknown microorganism. Final identiFi- cations labeled in the laboratory report as “very good “were considered accurate [23]. Preparation of Sabouraud dextrose broth (SDB) According to the manufacturer’s instruc- tions, 30 g of broth powder was dissolved in 1000 ml of distilled water and sterilized by autoclaving at 121°C and 15 psi for 15minutes. After sterilization, the broth was cooled to 47°C, and to prevent bacterial Effect of StronEum Titanate AddiEon on AnEfungal, Physical and Mechanical ProperEes of So?-Liners Vol 13, No 1 (2025) DOI 10.5195/d3000.2025.1014 http://dentistry3000.pitt.edu 4 growth, 0.05 g of chloramphenicol antibiotic was added [24]. Evaluating the effect of Chitosan / soft denture liner specimens on adherence of C. albicans adherence Sterile tubes were Filled with freshly pre- pared Sabouraud Dextrose Broth (SDB), into which a small amount of the isolated yeast culture was introduced. The yeast suspen- sion was then adjusted to a 0.5 McFarland standard using a McFarland densitometer. Pre-sterilized soft lining material specimens were immersed in the inoculated broth and incubated at room temperature for one hour. After incubation, the specimens were care- fully removed and gently rinsed with phos- phate-buffered saline (PBS) for one minute with mild rocking to remove loosely at- tached yeast cells, then dried using Filter pa- per. The adherent Candida cells were Fixed on the lining surface using methanol, stained with crystal violet for 60 seconds, rinsed again with PBS for 30 seconds, and Finally dried with Filter paper. Microscopic exami- nation was carried out using an inverted light microscope [25]. For each specimen, the attached Candida cells were counted in two standardized microscopic Fields, and the mean value of the two counts was calculated and recorded. Results and Discussion Scanning electron microscope (SEM) (Fig- ure 1) Results demonstrated that the incorporation of SrTiO₃ at concentrations of 1% and 1.5% contributed to improved surface morphol- ogy and reduced porosity. The nanoparticles exhibited good dispersion at the lower con- centration, while slight agglomeration was observed at the higher concentration. These surface modiFications may enhance the bond strength between the soft liner and the den- ture base, as well as improve mechanical properties such as hardness and resistance to fungal adhesion. Figure 1. SEM images of soft liner (con- trol,1%1.5%). Candida albicans Adherence The results of the multiple comparison table using the Games-Howell test designated sig- niFicant differences between group means (p < 0.05), conFirming the heterogeneous vari- ances. This demonstrated a clear effect of Strontium Titanate addition on the soft liner in comparison with the control group. Among the tested groups, Nystatin exhibited the greatest antifungal efFicacy, followed by 1.5% and then 1% Strontium Titanate, all of which signiFicantly outperformed the con- trol in reducing Candida growth (Figure 2). Figure 2. Candida albicans adherence. Shore A Hardness The source of the difference was more inves- tigated by the analysis of the data, tukey (Fig- ure 3). Through the mean differences, we noted the superiority of the group followed by 1.5 and then 1% over the control group in increasing hardness. Figure 3. Differences in hardness. Shear Bond Strength Statistically signiFicant changes in shear strength of bonding between the control group and the groups treated with either 1% or 1.5% Strontium Titanate was observed, according to Tukey's post hoc analysis. The Findings indicate that both concentrations improved shear bond strength compared to the control, with the 1% concentration demonstrating the highest mean value, out- performing even the 1.5% group (Figure 4). Figure 4. Differences in shear bond strength. Conclusion Within the parameters of this study, it was shown that addition silicone impression ma- terial can be safely disinfected by immersion in TTO for ten minutes without affecting the details of the reproduction and dimensional accuracy of the impressions. References 1. Elmwafy DA, Abdelghany AM. Stron- tium titanate nanoparticles modiEied denture base materials. Egypt Dent J. 2024 Jan;70(1):403–11. doi:10.21608/edj.2023.238541.2725 2. Mohammed Ali B. Preparation and eval- uation of some properties of heat-cured, acrylic- based soft denture liner. [PhD thesis]. Baghdad: University of Baghdad; 2006. 3. Shankargouda S. To evaluate different soft tissue liners, as functional impression materi- als, by determining its effect on surface roughness of dental stone over a time period: An in-vitro study. Mod Appr Dent Oral Health Care. 2018;3(2):247–55. 4. Hussein BM, Salem SA, Aliwi SM. Prepa- ration and evaluation of some properties of heat cured acrylic based denture soft liner. J Bag Coll Dent. 2015;27(4):32–6. CONTROL OL 1 % 1.5 % 0.1252 0.0321 0.0665 0.0642 0 0.05 0.1 0.15 0.2 control nystatin 1.00% 1.50% Candida albicans adherence test groups 15.28 21.21 22.42 0 10 20 30 control 1% 1.50% Shore A hardness test groups 0.7959 1.1418 1.0195 0 0.5 1 1.5 control 1% 1.50% Shear bond strength test groups Effect of StronEum Titanate AddiEon on AnEfungal, Physical and Mechanical ProperEes of So?-Liners Vol 13, No 1 (2025) DOI 10.5195/d3000.2025.1014 http://dentistry3000.pitt.edu 5 5. Kreve S, Dos Reis AC. Denture liners: A systematic review relative to adhesion and me- chanical properties. 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Philadelphia: Lip- pincott Williams & Wilkins; 2001 24. Forbes BA, Sahm DF, Weissfeld AS, Bai- ley SS. Diagnostic microbiology. 12th ed. St. Louis: Mosby Elsevier; 2007. 25. Chander J. Textbook of medical mycol- ogy. London: JP Medical Ltd; 2017. 26. Byadarahally RS, Rajappa S. Isolation and identiEication of Candida from the oral cavity. Int Scholarly Res Netw Dent. 2011;2011:1-6. Table 1. The mixing ratios of SrTio3 nanoparticles, Soft Liner(PMMA)(MMA), Nystatin. Test Selected concen- tration % Amount of SRTI O3 powder (g) Amount of polymer powder (g) Amount of mono- mer liquid (ml) Shear Control 0 10 8.1 1 0.1 9.9 8.1 1.5 0.15 9.85 8.1 Hardness Control 0 10 8.1 1 0.1 9.9 8.1 1.5 0.15 9.85 8.1 Candida albicans Control 0 10 8.1 Control + 1.wt.% nystatin 1.4 8.6 8.1 1% 0.1 9.9 8.1 1.5% 0.15 9.85 8.1