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ORIGINAL ARTICLE
Year : 2020  |  Volume : 15  |  Issue : 1  |  Page : 63-67

Evaluation of antifungal effect of maxillofacial silicone after incorporation of chitosan nanoparticles: Evidence in pharmaceutical therapeutics


Department of Prosthodontics, Sharad Pawar Dental College & Hospital, Datta Meghe Institute of Medical Sciences (Deemed to be University), Sawangi (M), Wardha, Maharashtra, India

Date of Submission23-Feb-2020
Date of Decision10-Mar-2020
Date of Acceptance23-Mar-2020
Date of Web Publication13-Oct-2020

Correspondence Address:
Dr. Tanvi Jaiswal
Department of Prosthodontics, SPDC, Sawangi (M), Wardha - 442 001, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jdmimsu.jdmimsu_51_20

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  Abstract 


Background: The incorporation of nanotechnology in dentistry is evident, especially in the past decade. The pharmaceutical research in nanotechnology has been a most important invention while considering its application in dental materials. The evidence gathered from the various pharmacotherapeutic researches on the incorporation of chitosan nanoparticles (NPs) to improve the antimicrobial activity of various dental materials are proved, although there is very less evidence of use of functionalized materials to improve its efficacy in clinical situations. Aim: The aim of the study was to evaluate the antifungal effect of maxillofacial silicone after incorporation of chitosan NPs in patients with maxillary defect. Objectives: The objective of the study was to compare the effectiveness of antifungal activity of silicone by incorporating chitosan NPs to that of the control group at various time intervals. Materials and Methods: Chitosan NPs were incorporated in silicone material for relining of interim obturator to check its efficacy with that of conventional interim obturator. Results: Experimental group of patients in which obturator is relined with silicone material consists chitosan NPs has shown to be more effective against fungal colonization growth as compared to that of control group. Conclusion: Chitosan NP with silicone maxillofacial material proved its efficacy against candida growth in patients of maxillary defects at various time intervals.

Keywords: Antifungal agent, chitosan nanoparticles, pharmacotherapeutics


How to cite this article:
Jaiswal T, Pisulkar SK, Kambala SS. Evaluation of antifungal effect of maxillofacial silicone after incorporation of chitosan nanoparticles: Evidence in pharmaceutical therapeutics. J Datta Meghe Inst Med Sci Univ 2020;15:63-7

How to cite this URL:
Jaiswal T, Pisulkar SK, Kambala SS. Evaluation of antifungal effect of maxillofacial silicone after incorporation of chitosan nanoparticles: Evidence in pharmaceutical therapeutics. J Datta Meghe Inst Med Sci Univ [serial online] 2020 [cited 2020 Oct 28];15:63-7. Available from: http://www.journaldmims.com/text.asp?2020/15/1/63/297994




  Introduction Top


The research in the field of nanotechnology has been a boon to various pharmaceutical drug delivery issues and therapeutic evidences. Nanoparticles (NPs) are the most useful modalities in modern pharmaceutical and biomedical fields. Pharmaceutical nanotechnology embraces the application of nanoscience to the pharmacy as a nanomaterial and as devices such as drug delivery, diagnostic, imaging, and biosensor.[1] Oral cancer is one of the most dreadful and life-threatening diseases in today's world. The various factors affecting the prevalence of oral cancer differ from country to country and also vary depending on age, lifestyle, and the socioeconomic status of an individual.[2] The postsurgical malformations in the head and neck resulting postsurgical treatment of either benign or malignant neoplasm or trauma except for patients may be amenable to surgical closure.[3] Patients with such defects are rehabilitated by prosthodontic means or artificial repair with maxillofacial prosthesis. The obturators provided in the form of maxillofacial prosthesis are fabricated with silicone that is always in contact with soft tissues, saliva, and nasal secretions. Constant exposure to these body fluids might further lead to colonization of various pathogenic microorganisms over their surfaces causing elastomer degradation or infection,[4] which ultimately results in an increased percentage of consumption of drugs, thereby leading to formation of drug resistance by particular microorganisms. There are various forms of generic drugs that have been proved very efficient in controlling the fungal infections of the oral cavity. Although invariably being considered the medicine of choice against fungal growth, particularly Candida albicans, various antifungal agents are increasingly reported as inefficient on account of number of cases reported with resistances, particularly to fluconazole. This endemic thereby evolves a need for invention for alternative modalities incorporated for the management of C. albicans infections, especially in the oral cavity.[5],[6] In the recent era, nanoscience and nanotechnology are gaining tremendous popularity. The various applications of NPs in various fields have proved as a novel alternative to overcome resistance toward bacterial and fungal drugs reported globally due to the misuse of antibiotics.[7] In recent years, chitosan is considered as effective biopolymers for biomedical applications. Chitosan is cellulose-like naturally derived polysaccharide (biopolymer), whereby various alkaline substances such as sodium hydroxide treat the chitin shells of crustaceans, which is also a biodegradable, biocompatible, nontoxic, and hydrating agent consisting of antimicrobial properties. Chitosan by virtue of its amino groups is the only natural polysaccharide with cationic character. Chitosan is protonated and can interact with negatively charged compounds, such as proteins, anionic polysaccharides (e.g., alginates, carraghenates, and pectins), fatty acids, bile acids, and phospholipids at low pH. This property, along with its biocompatibility, biodegradability, and lack of toxicity, has led to the usage of chitosan in diverse fields, such as technology, food, cosmetics, medicine, biotechnology, agriculture, and the paper industry.[5] Various evidence-based literature and reports suggest that chitosan is active on C. albicans, especially when incorporated in prosthetic materials, although all such studies carried out are in vitro studies and evidence toward the implication of chitosan in maxillofacial prosthetic materials and its activity in clinical situations are still lacking, which may have a wide future scope.

Thereby considering this aspect, the present research is an attempt toward developing a new silicone material with chitosan NPs through the functionalization of medical-grade silicone material (Factor II-2186) in the patients with maxillary defect and also evaluated its efficacy in various time intervals.

Aim

The aim of the study was to evaluate the antifungal effect of maxillofacial silicone after incorporation of chitosan NPs in patients with maxillary defect.

Objectives

  • To evaluate the antifungal activity of interim obturator without any intervention (control group), in interval of 2 weeks, 4 weeks postinsertion of the interim obturator.
  • To evaluate the antifungal activity of silicone after incorporating chitosan NPs, in interval of 2 weeks, 4 weeks postinsertion of the interim obturator
  • To compare the effectiveness of antifungal activity of silicone by incorporating chitosan NPs to that of the control group at various time intervals.



  Materials and Methods Top


Materials

Room-temperature-vulcanized silicone elastomer A-2186 (Factor II) was used in this study.

Chitosan NPs were obtained from Nanoshel, Intelligent Materials Pvt., Ltd.

Methodology

An interventional pilot study was carried out in the Department of Prosthodontics, Sharad Pawar Dental College, Sawangi (Meghe), DMIMS (DU), Wardha, after the ethical clearance from the Institutional Ethical Committee (DMIMS (DU)/IEC/January 2019/8105). Six patients undergoing a surgical intervention of maxilla (developmental or acquired defect), after which obturator as rehabilitation prosthesis was the treatment of choice, were selected for the study. The present study specifically focused on interim obturators. A total number of patients randomly were selected in two groups of which Group A is the control group and Group B is an Experimental group with chitosan NPs.

In Group A that was the control group of patients, after 1 month of surgery, interim obturator will be fabricated in conventional manner relined with silicone and patients were recalled every after 2 weeks of insertion for an alteration of prosthesis, and at every visits (i.e.,: 0, 2, and 4 weeks), swabs were made from defect site and from intaglio surface of the obturator with paper points to check microbial colonization growth.

Time log for swab: Group A

Time log for swab in Group A included AT0 – at the time of insertion, AT1 – after 2 weeks, and AT2 – after 4 weeks.

In Group B that was the experimental group of patients, a new impression of defect site was made and the obturator was fabricated; obturator was relined with silicone material consisting of chitosan NPs. Patients were recalled every after 2 weeks of insertion for follow-up and alteration of prosthesis, and at every visits (i.e.,: 0, 2, and 4 weeks), swabs were made from defect site and from intaglio surface of obturator with paper points to check microbial colonization growth.

Time log for swab: For Group B

Time log for swab for Group B included BT0 – at the time of insertion, BT1 – after 2 weeks, and BT2 – after 4 weeks.

For each patient, sabouraud dextrose agar (SDA) was inoculated to check for the fungal growth and the medium was incubated at 37°C for 48 h. After inoculation and time of incubation period, the specimens were removed using sterile forceps to avoid any cross-contamination. Growth on SDA within 24–48 h at 37°C was identified and confirmed as Candida species.

  • Growth of colonies on each  Petri dish More Details was counted using colony counter to identify the number of colonies and the growth on control [Figure 1], [Figure 2], [Figure 3] and experimental groups [Figure 4], [Figure 5], [Figure 6]
  • To perform Gram staining, new slides were selected for each patient. Each slide was made grease-free by washing with soap and water. Growth of colonies was collected from the medium rolled on slides with inoculating loop; they were rapidly passed over a flame of Bunsen burner 2–3 times for heat fixing
  • Gram-positive approximately 4–8 μm dark purple yeast cells with budding were considered positive for C. albicans [Figure 7].



  Observation and Results Top


Statistical analysis was carried out using descriptive and inferential statistics applying student's paired and unpaired t-test. Software used in the analysis was SPSS24.0 version (Chicago, Illinois, USA)and P < 0.05 was considered as a level of significance. Comparative evaluation of the results of both the groups shows a statistically significant difference between the colony-forming units of control and experimental groups at various time intervals. Experimental group of patients where the obturator is relined with silicone material consisting of chitosan NPs has shown to be more effective against fungal colonization growth as compared to that of the control group [Table 1] and [Graph 1].
Figure 1: Group A: AT0 – At time of insertion

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Figure 2: Group A: AT1 – After 2 weeks

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Figure 3: Group A: AT2 – After 4 weeks

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Figure 4: Group B: BT0 – At time of insertion

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Figure 5: Group B: BT1 – After 2 weeks

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Figure 6: Group B: BT2 – After 4 weeks

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Figure 7: Gram-positive dark purple yeast cells with budding, suggestive of Candida albicans

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Table 1: Table of comparison of growth of colonies in two groups (Group A and Group B)

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  Discussion Top


Oral cancer is considered a serious public health problem that causes a great deal of morbidity and mortality in the population. It is often painless and asymptomatic, and thus, the patient usually reports at the advanced stage of the disease for which surgical excision is the only treatment option remains followed by the prosthesis.[8]

Maxillofacial prosthesis is designed to replace lost facial and intraoral parts of the defect site. Retention, stability, and functioning of prosthesis are quite questionable in patients with large defects. There are many options for providing retention, but all of them causing somewhat discomfort to patient while retrieving the prosthesis or in other ways during function. Natarajan et al. in 2013 in their case reported utilization of silicone as a relining material to engage the undercut and it relieves pressure during function. It aids in relatively better retention and stability, which a rigid obturator fails to provide. Based on the evidence of utilization in this study, we have used silicone material to reline interim obturator, and our study supports the thought of Natarajan et al.[9] in maintaining its function. Prosthesis designed in this manner provides a better functional outcome to a compromised clinical situation. Adequate wettability is one significant feature that a maxillofacial prosthetic material which contacts the skin or mucosa should possess. A uniform wetted surface will lead to a superior lubricating layer between supporting tissues, thereby reducing friction and patient discomfort. Another prominent quality is the surface energy of a maxillofacial prosthetic material that provides an indication of amount of energy available for adhesion and of susceptibility of the material to bacterial adhesion.[10] Contamination and infection by virtue of continuous salivary contact and exposure to nasal fluids is more prone in maxillofacial prosthetic materials. A significant challenge for prostheses fabricated with silicone material, especially in such clinical scenarios, is the prevalence of fungal infections leading to candida colonization. Exposure to various medical risks resulting as a sequel of fungal infections is higher in maxillofacial prosthesis wearer due to candida's adherence on the material surface. Candida causes degradation of the material and infection of the surrounding tissue resulting in a complex biofilm formation on the prosthesis.[11] The present study favored the same facts as suggests by Al-Hakam J Ibrahim that maxillofacial silicone prosthesis causes more adherence to fungal or candida infections.

Recently, as NPs have emerged as a breakthrough in the form of a novel alternative in the field of pharmacological advances to overcome multidrug resistance toward microbial infections encountered on a global level due to misuse of antibiotics. Incorporation and addition of nanomaterials have time and again proven their efficacy in broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria, mycobacteria, and fungi. Chitosan and its derivatives have attracted considerable interest due to their antimicrobial and antifungal activity as proven in vitro. Chitosan exhibits its activity only in an acidic medium because of its poor solubility >pH 6.5, although there is a lack of clinical research in a combination of chitosan with maxillofacial silicone materials carried out in the past.[12] By considering above all facts and evidences of earlier in vitro studies, the present study is carried out in which chitosan is incorporated in maxillofacial silicone material to formulate it as antifungal material for fabricating maxillofacial prosthesis. The results of the present study conclude the fact that Group B that is experimental group of patients in which obturator is relined with silicone material consist chitosan NPs has shown to be more effective against fungal colonization growth as compared to that of Group A that is control group of patients in which interim obturator was fabricated in a conventional manner [Table 1] and Graph 1]. The justification for this observation can be credited to the wide range of antimicrobial properties of chitosan NPs by virtue of its incorporation in the silicone. These results of the study formulated a new silicone maxillofacial material, which would be more effective for the fabrication of obturator with antifungal activity in future.


  Conclusion Top


After comparatively evaluating the results of this study for the growth of surface colonization of candida after incorporation of chitosan NPs, it was concluded that the results of viable count proved a highly significant reduction in colony-forming unit of C. albicans in experimental group in comparison with control group, which increases the efficacy of this functionalized medical-grade silicone that is Factor II A 2186. Chitosan and its derivatives possess very attractive biological activities. Countless researches have been carried out and have commonly reported excellent activities without toxicity. Their unique and attractive bioactivities have been proved.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Gadad AP, Kumar SV, Dandagi PM, Bolmol UB, Pallavi NP. Nanoparticles and their therapeutic applications in pharmacy. Int J Pharm Sci Nanotechnol 2014;7:2509-019.  Back to cited text no. 1
    
2.
Kadashetti V, Shivakumar KM, Chaudhary M, Patil S, Gawande M, Hande A. Influence of risk factors on patients suffering from potentially malignant disorders and oral cancer: A case-control study. J Oral Maxillofac Pathol 2017;21:455-6.  Back to cited text no. 2
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Al-Hakam JB, Al-Judy HJ. Mechanical properties of chitosan incorporated in maxillofacial silicone and its anti candidal activity in vitro. J Res Med Dent Sci 2018;6:101-7.  Back to cited text no. 3
    
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Kurunmäki H, Kantola R, Hatamleh MM, Watts DC, Vallittu PK. A fiber-reinforced composite prosthesis restoring a lateral midfacial defect: A clinical report. J Prosthet Dent 2008;100:348-52.  Back to cited text no. 4
    
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Costa E, Silva S, Tavaria F, Pintado M. Antimicrobial and antibiofilm activity of chitosan on the oral pathogen Candida albicans. Pathogens 2014;3:908-19.  Back to cited text no. 5
    
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Seyfarth F, Schliemann S, Elsner P, Hipler UC. Antifungal effect of high- and low-molecular-weight chitosan hydrochloride, carboxymethyl chitosan, chitosan oligosaccharide and N-acetyl-D-glucosamine against Candida albicans, Candida krusei and Candida glabrata. Int J Pharm 2008;353:139-48.  Back to cited text no. 6
    
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Fernando SS, Gunasekara TD, Holton J. Antimicrobial nanoparticles: Applications and mechanisms of action. Sri lankan J infect Dis 2018;8:2-11.  Back to cited text no. 7
    
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Dangore-Khasbage AS. Clinical aspects of oral cancer: A case report series Kliniczne uwarunkowania raka jamy ustnejopis przypadków. 2017, vol. 54, nr 1, January-March, p. 85-9. doi: 10.17219/dmp/67499.  Back to cited text no. 8
    
9.
Natarajan P, Ramasamy C, Padmanabhan TV. Rehabilitation of a partial maxillectomy patient with silicone relined hollow bulb obturator–Case report. IOSR Journal of Dental and Medical Sciences (IOSR-JDMS). 2013;11:1-5.  Back to cited text no. 9
    
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Waters MG, Jagger RG, Polyzois GL. Wettability of silicone rubber maxillofacial prosthetic materials. J Prosthet Dent 1999;81:439-43.  Back to cited text no. 10
    
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Jadhav RD, Bhide SV, Prasad BL, Shimpi J. Applications of silver nanoparticles in prosthodontics: An overview. Europ J Prosthod 2016;4:45.  Back to cited text no. 11
    
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Qi L, Xu Z, Jiang X, Hu C, Zou X. Preparation and antibacterial activity of chitosan nanoparticles. Carbohydr Res 2004;339:2693-700.  Back to cited text no. 12
    


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  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]
 
 
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