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 Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 13  |  Issue : 5  |  Page : 124-127  

Antibacterial and antifungal efficacy of platelet-rich fibrin and platelet-rich fibrin matrix against root canal microflora


1 Department of Dentistry, Patna Medical College and Hospital, Patna, Bihar, India
2 Department of Conservative Dentistry and Endodontics, Narayan Swami Hospital and Dental College, Dehradun, Uttarakhand, India
3 Department of Conservative Dentistry and Endodontics, Maharana Pratap Dental College, Kanpur, Uttar Pradesh, India
4 Department of Public Health Dentistry, Dr. Rajesh Ramdasji Kambe Dental College and Hospital, Akola, Maharashtra, India
5 Private Practitioner, Patna, Bihar, India
6 Department of Conservative Dentistry and Endodontics, RKDF Dental College and Research Center, Bhopal, Madhya Pradesh, India

Date of Submission27-Sep-2020
Date of Acceptance28-Sep-2020
Date of Web Publication05-Jun-2021

Correspondence Address:
Pinky Singh
Patna Medical College and Hospital, Patna, Bihar
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jpbs.JPBS_601_20

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   Abstract 


Background: Binding of such proteins with a developing fibrin mesh or to the extracellular matrix has the ability to create chemotactic gradients aiding the recruitment of stem cells, thereby, stimulating cell migration and differentiation this may promotes repair and regeneration. There is a unique concept in platelet concentrates called platelet-rich fibrin matrix (PRFM) which is processed using higher attraction without the utilization of bovine thrombin. Aim and Objectives: The aim of this study is to evaluate the antibacterial and antifungal effects of platelet-rich fibrin (PRF) and PRFM against root canal microflora. Materials and Methods: Blood samples were taken from 20 adults, age ranging from 20 to 40 years who were systemically healthy, nonsmokers with no symptoms of infection, or on antibiotics at least 3 months before experiments. The procedures and the protocols involved in the study were accepted by the Institutional Ethical Committee. Written informed consent was obtained from the donors and patients. Approximately 10 ml of blood was collected from each donor. 5 ml of blood was used for the preparation of PRF and 5 ml for PRFM. Results: The antifungal scores among the groups. The highest antifungal scores were seen in metapex followed by PRF and PRFM group. Kruskal–Wallis test was applied to check the statistical difference among the groups, and there was a statistically significant difference seen among the groups (P = 0.00). The antibacterial scores among the groups. The highest antibacterial scores were seen in Metapex followed by PRF group and PRFM group Kruskal–Wallis test was applied to check the statistical difference among the groups, and there was a statistically significant difference seen among the groups. Conclusion: PRF demonstrated antibacterial activity against passage isolates but had no antifungal efficacy. This antibacterial property may be a valuable adjunct when using PRF for regenerative procedures in endodontics. Both PRF and PRFM have to be analyzed for antimicrobial properties for extended duration and various other clinical strains with larger sample size.

Keywords: Aerobic, anaerobic, Gram-positive, immunologic response, platelet concentrates, regenerative procedures


How to cite this article:
Singh P, Dey S, Pandey V, Abhas A, Sharan S, Kharat N. Antibacterial and antifungal efficacy of platelet-rich fibrin and platelet-rich fibrin matrix against root canal microflora. J Pharm Bioall Sci 2021;13, Suppl S1:124-7

How to cite this URL:
Singh P, Dey S, Pandey V, Abhas A, Sharan S, Kharat N. Antibacterial and antifungal efficacy of platelet-rich fibrin and platelet-rich fibrin matrix against root canal microflora. J Pharm Bioall Sci [serial online] 2021 [cited 2021 Oct 27];13, Suppl S1:124-7. Available from: https://www.jpbsonline.org/text.asp?2021/13/5/124/317575




   Introduction Top


Endodontic regenerative procedures are biologically based procedures which have regeneration of pulp-like tissue, more idealistically the dentin-pulp complex, affected coronal dentin following a carious exposure or trauma, and regeneration of pathological root resorption in cervical, middle, or apical areas.[1] An increased understanding of the physiological therapeutic role of platelets in wound healing has led to the employment of platelet concentrates (PCs) as remedial tools for regenerative protocols.[2] It contains platelets and growth factors (GFs) within the kind of fibrin membrane. It is prepared from the patient's own blood and is free from any anticoagulant. These PCs contain biologically active protein that accelerates the wound healing, and additionally, promotes a angiogenesis, tissue repair, however, causes moderate inflammation and an immunologic response.[3] The binding of such proteins with a developing fibrin mesh or to the extracellular matrix has the ability to create chemotactic gradients aiding the recruitment of stem cells, thereby, stimulating cell migration and differentiation this may promotes repair and regeneration. There is a unique concept in PCs called platelet-rich fibrin matrix (PRFM) which is processed using higher attraction without the utilization of bovine thrombin.[4] It is obtained after two sets of centrifugation and is polymerized using calcium chloride.

Microorganisms that ends up in endodontic infections are mainly of low virulence. Their pathogenesis and endurance are influenced by the discharge of lipopolysaccharide, toxins, and therefore, the synthesis of enzymes. Pulpal and periapical lesions are related to a mixed microbiota, that consists of aerobic, anaerobic, Gram-positive, and negative microorganisms.[5] In the literature, the application of platelet-rich fibrin (PRF) as a potential scaffold for regenerative endodontic protocol has been documented. Literature survey reveals limited studies assessing antibacterial and antifungal properties of PRF and PRFM for regenerative endodontics. Hence, the aim of the study was to evaluate the antibacterial and antifungal effects of PRF and PRFM against root canal microflora.


   Materials and Methods Top


Blood samples were taken from 20 adults, age ranging from 20 to 40 years who were systemically healthy, nonsmokers with no symptoms of infection or on antibiotics at least 3 months before experiments. The procedures and the protocols involved in the study were accepted by the Institutional Ethical Committee. Written informed consent was obtained from the donors and patients. Approximately 10 ml of blood was collected from each donor. 5 ml of blood was used for the preparation of PRF and 5 ml for PRFM.

Samples of PRF and PRFM divided into two groups, and they are as follows:

  • Anti-bacterial
  • Antifungal.


Ten samples of PRF and PRFM were used antibacterial group and 10 samples of PRF and PRFM were used in anti-fungal group. Metapex was used in the control group.

Statistical analysis

The mean value of zone of inhibition was observed in relation to PRF, PRFM, and Metapex, and statistical analysis was done using the SPSS software version 20 (IBM Co. , California, USA). The mean values were analyzed using the Kruskal–Wallis test, and Mann–Whitney test was applied to check the statistical difference between the groups.


   Results Top


[Table 1] shows the antifungal scores among the groups. The highest antifungal scores were seen in metapex followed by PRF and PRFM group. Kruskal–Wallis test was applied to check the statistical difference among the groups, and there was a statistically significant difference seen among the groups (P = 0.00) [Graph 1].
Table 1: Comparison of the mean distribution of the antifungal scores among the groups

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[Table 2] shows the antibacterial scores among the groups. The highest antibacterial scores were seen in metapex followed by PRF group and PRFM group. Kruskal–Wallis test was applied to check the statistical difference among the groups, and there was a statistically significant difference seen among the groups (P = 0.00).
Table 2: Comparison of the mean distribution of the antibacterial scores among the groups

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For post hoc, Mann–Whitney test was applied to check the statistical difference between the groups, as shown in [Table 3]. There was a statistical significant difference seen between PRF and PRFM, PRF and metapex, PRFM and metapex with respect to antifungal and antibacterial scores (P ≤ 0.05).
Table 3: Post hoc

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   Discussion Top


Platelets also participate in antibody-dependent cell cytotoxicity functions to destroy protozoal pathogens by which platelets release an array of potent antimicrobial peptides.[6],[7] Activated platelets release various GFs and platelet microbicidal proteins,[8] which may be a mixture of platelets, leukocytes, and plasma. The plasma contains complement, which is an integral variable of the system, plays role within the antimicrobial activity. The leukocyte concentration and handling characteristics of PRF and PRFM differ and PRFM lacks leukocytes. Leukocytes have an intrinsic role in antimicrobial potential and reaction. Insignificant antimicrobial activity of PRFM may attribute to the absence of leukocytes. Bielecki et al. did a study on the impact of leukocyte in PCs and their role in immune reaction and wound healing. Elements of neutrophils such as polymorphonuclear neutrophilic granulocytes granule proteins, cathepsin G, heparin-binding protein, calprotectin, defensins, phospholipase A2, and eosinophils are effective immune mediators.[9] The leukocyte concentration and handling characteristics of PRF and PRFM differs, PRFM lacks leukocytes. Leukocytes have an intrinsic role in antimicrobial potential and reaction. Insignificant antimicrobial activity of PRFM may attribute to the absence of leukocytes. Bielecki et al. did a study on the impact of leukocyte in PCs and their role in immune reaction and wound healing. The components of neutrophils such as polymorphonuclear neutrophilic granulocytes, cathepsin G, defensins, phospholipase A2, heparin-binding protein, Calprotectin, and eosinophils are effective immune mediators.[9] The increased rate of protease activity of candida compared with other candida species also suggests its higher virulence characteristics. Candida albicans has the potential to grow on the dentinal surfaces even within the absence of oral tissue fluids and penetrates into the dentinal tubules by its own various growth forms.[10] Sen et al. suggested that Candida is taken into consideration as dentinophilic microorganism.[11] It's for this reason that the sample within this study was obtained by filing the dentinal walls of the canals using K file. The results of our study showed that caustic lime was more efficient on C. albicans as compared to the PRF and PRFM [shown in [Graph 2]]. These results can be attributed to its ability to inactivate the cytotoxic effects of the endotoxin released by the microorganisms. The second reason could due to the usage of hydrated lime; however, PRF and PRFM failed to show any antifungal properties.




   Conclusion Top


PRF demonstrated antibacterial activity against passage isolates but had no antifungal efficacy. This antibacterial property may be a valuable adjunct when using PRF for regenerative procedures in endodontics. Both PRF and PRFM have to be analyzed for antimicrobial properties for extended duration and various other clinical strains with larger sample size.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Murray PE, Garcia-Godoy F, Hargreaves KM. Regenerative endodontics: A review of current status and a call for action. J Endod 2007;33:377-90.  Back to cited text no. 1
    
2.
Naik B, Karunakar P, Jayadev M, Marshal VR. Role of platelet rich fibrin in wound healing: A critical review. J Conserv Dent 2013;16:284-93.  Back to cited text no. 2
[PUBMED]  [Full text]  
3.
Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part IV: Clinical effects on tissue healing. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006;101:e56-60.  Back to cited text no. 3
    
4.
Fouad AF, Verma P. Healing after regenerative procedures with and without pulpal infection. J Endod 2014;40:S58-64.  Back to cited text no. 4
    
5.
de Mendonça Cavalcante A, Soares NM, Santos IC, de Azevedo Ximenes EA, da Silva MA, Junior KA. Assessment of microbiota in root canals with pulp necrosis by means of Gram test. Afr J Microbiol Res 2018;12:508-11.  Back to cited text no. 5
    
6.
Klinger MH, Jelkmann W. Role of blood platelets in infection and inflammation. J Interferon Cytokine Res 2002;22:913-22.  Back to cited text no. 6
    
7.
Tang YQ, Yeaman MR, Selsted ME. Antimicrobial peptides from human platelets. Infect Immun 2002;70:6524-33.  Back to cited text no. 7
    
8.
Yeaman MR, Tang YQ, Shen AJ, Bayer AS, Selsted ME. Purification and in vitro activities of rabbit platelet microbicidal proteins. Infect Immun 1997;65:1023-31.  Back to cited text no. 8
    
9.
Bielecki T, Dohan Ehrenfest DM, Everts PA, Wiczkowski A. The role of leukocytes from L-PRP/L-PRF in wound healing and immune defense: New perspectives. Curr Pharm Biotechnol 2012;13:1153-62.  Back to cited text no. 9
    
10.
Chandra SS, Miglani R, Srinivasan MR, Indira R. Antifungal efficacy of 5.25% sodium hypochlorite, 2% chlorhexidine gluconate, and 17% EDTA with and without an antifungal agent. J Endod 2010;36:675-8.  Back to cited text no. 10
    
11.
Sen BH, Safavi KE, Spångberg LS. Growth patterns of Candida albicans in relation to radicular dentin. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1997;84:68-73.  Back to cited text no. 11
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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