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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 12  |  Issue : 2  |  Page : 59-62

Silver-impregnated platelet-rich fibrin as a barrier membrane


1 Department of Periodontology, MGVM K. B. H. Dental College and Hospital, Nashik, Maharashtra, India
2 Vinay Heart Clinic, Nashik, Maharashtra, India
3 Department of Periodontology, Y. M. T. Dental College and Hospital, Navi Mumbai, Maharashtra, India

Date of Submission30-Aug-2019
Date of Acceptance09-Jan-2020
Date of Web Publication22-Jul-2020

Correspondence Address:
Dipali Chaudhari
Department of Periodontology, MGVM K. B. H. Dental College and Hospital, Nashik, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jorr.jorr_32_19

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  Abstract 


Introduction: Platelet-rich fibrin (PRF) was developed to eliminate xenofactors form of platelet-rich plasma to be used as a source of growth factor for tissue regeneration. PRF has been used as an autologous grafting material because of its ability to accelerate physiologic wound healing and new bone formation.
Aim: To compare mechanical and histologic characteristics of the PRF membrane and silver-impregnated PRF.
Materials and Methods: Venous blood was taken from the subjects for PRF preparation. Then, 2 normal PRFs were prepared, and for silver-impregnated PRF, 9 ml of blood in addition to 1 ml of silver nanoparticle suspension was poured into another tube and gently shook with hand to achieve a uniform 1% concentration. The tubes were centrifuged at 3000 rpm for 10 min. The tensile test was done by the universal testing machine:

  1. The remaining pieces of the membranes were fixed in 10% formalin for 24 h to be subjected to hematoxylin and eosin staining and evaluated with the light microscope
  2. Degradation time.

Results: Silver-impregnated showed improved mechanical properties and dense fibrin network than PRF.
Conclusion: Silver-impregnated PRF membrane demonstrated properties to be used as a barrier membrane for periodontal reconstruction.

Keywords: Guided tissue regenaration, platelet-rich fibrin, silver nanoparticles


How to cite this article:
Chaudhari D, Mahale S, Mahale A, Warang A, Kalekar L, Shimpi S. Silver-impregnated platelet-rich fibrin as a barrier membrane. J Oral Res Rev 2020;12:59-62

How to cite this URL:
Chaudhari D, Mahale S, Mahale A, Warang A, Kalekar L, Shimpi S. Silver-impregnated platelet-rich fibrin as a barrier membrane. J Oral Res Rev [serial online] 2020 [cited 2020 Aug 13];12:59-62. Available from: http://www.jorr.org/text.asp?2020/12/2/59/290504




  Introduction Top


Guided tissue regeneration (GTR) and guided bone regeneration (GBR) are surgical techniques that aim to reconstruct the damaged periodontal tissues, which are lost due to periodontal lesions, and to regain the alveolar bone, which are lost due to tooth extraction or periodontal disease.[1] These methods employ various membranes to cover the bone and periodontal ligament and temporarily separate them from the epithelium and gingival connective tissue.[2] Regenerative potential of platelets was first introduced in the 1970s, just when they were found to contain growth factors responsible for increasing the collagen production, cell mitosis, blood vessels growth, and induction of cell differentiation.[3] The platelets were increasingly used in tissue regeneration over time. The platelet-rich fibrin (PRF) can be used in various regenerative treatments to accelerate the healing and improve the regeneration procedure.[4] It can also be used as a scaffold in tissue engineering.[5] At present, there are several techniques to obtain the high concentration of platelets, each of which results in a specific product that is unique in terms of biology and performance. These methods are generally classified into four groups based on their fibrin and leukocyte content: pure PRP, leukocyte- and PRP, pure P-PRF, and leukocyte- and PRF.[6]

The chemical and physical properties of the membrane can influence the ultimate outcome of GBR and GTR.[7] The tensile strength of the tissue or the material, which is sutured, affects the success of suturing and the clinical results of wound healing.[8] Meanwhile, the membrane stiffness and the presence of stiff material influence the distribution of mechanical forces over the surrounding tissues.[9] Generally, the better the mechanical properties of the membrane provide the better support for regenerative treatments.

The most frequent postoperative complication of different regenerative techniques is the membrane exposure to the oral cavity, in which case, oral cavity microorganisms can colonize on the membrane and jeopardize the success of treatment.[10] It results in higher risk of infection and poor bone healing even in healthy individuals. Reinforcing the membranes' antimicrobial properties with inorganic materials can improve the treatment results. Inorganic antimicrobial materials have been more appreciated recently due to their safety and stability. One of the substances widely used today in various medical fields is silver nanoparticle (SNP). SNPs released up to 90% of their weight into human tissue, but the dissolution was never completed. SNPs have the ability to anchor to the bacterial cell wall and penetrate in it and cause structural changes in the cell membrane affecting its permeability and cell death. It causes formation of free radicals which lead to cell death.

It has been shown that these particles have high biocompatibility and also have favorable properties, including antimicrobial properties.[11] Studies reported the effect of these materials on a wide spectrum of Gram-negative and Gram-positive bacterial as well as antibiotic-resistant species. In addition, their antifungal and antiviral effects were proven.[10] The purpose of this study was to compare the mechanical and histologic characteristics of the PRF membrane before and after the addition of SNPs.


  Materials and Methods Top


Platelet-rich fibrin preparation

Details of the study design and consent form were approved by the Ethical Committee (Institutional ethical committee MGVM KBH dental college MGV/KBHDC/790/2017-18). Nine milliliters of the blood was collected from healthy, nonsmoking volunteer aged 28 years (male) using Vacutainer™ tubes and immediately centrifuged by a Medifuge centrifugation system at 3000 rpm for 10 min After the red thrombus (fraction of red blood cells) was eliminated from the PRF preparations, the resulting PRF was compressed with dry gauze for 15 s.

Preparing silver nanoparticle suspension[12]

To obtain a uniform suspension, 0.1 g nanosilver powder [Figure 1] with particles sized <100 nm (Sigma Aldrich, USA), along with 1 cc normal saline, was poured into the tube and sonicated at 200 W for 2 min in a sonicator device.
Figure 1: Silver nanoparticles

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Silver nanoparticle-impregnated platelet-rich fibrin membrane[12]

The remaining 9 ml in addition to 1 ml of SNP suspension was poured into another tube and gently shook with hand to achieve a uniform 1% concentration and then centrifuged by a Medifuge centrifugation system at 3000 rpm for 10 min. After the red thrombus (fraction of red blood cells) was eliminated from the PRF preparations, the resulting SNP-impregnated PRF was obtained [Figure 2].
Figure 2: Silver-impregnated platelet-rich fibrin

Click here to view


Histological processing[13]

Both PRFs were fixed in 10% neutralized formalin, dehydrated, and embedded in paraffin block sectioned sagittally. Sections were stained with hematoxylin and eosin for the demonstration of nucleus and cytoplasmic inclusions in specimens.

Physical properties[12]

Tensile strength was measured using universal testing machine [Figure 3]. Sufficient sustained tensile strength to avoid membrane collapse and function as a barrier. The ideal membrane should be sufficiently rigid to withstand the compression of overlying soft tissues and possess the required degree of plasticity for being easily contoured and molded into the desired shape to conform to the defect. The “universal” part of the name reflects that it can perform many standard tensile and compression tests on materials, components, and structures. The specimen was placed in the machine between the grips and an extensometer. Once the machine was started, it begins to apply an increasing load on specimen. Throughout the tests, the control system and its associated software recorded the load and extension or compression of the specimen.
Figure 3: Universal tensile machine

Click here to view


Accelerated degradation in vitro test[13]

Two types of PRF membrane disks were freshly prepared, were inserted in 24-well plates, and were incubated in a CO2 incubator with Hank's balanced salt solution (HBSS) supplemented with human plasmin (2.l g/ml).[1] HBSS was added to each well (24-well plate) by 0.5 ml and changed every 2 days.


  Results Top


Histological characteristics

Evaluating the microscopic sections, the SNPs were observed all over the membrane, but in the outer layers, they were more densely attached to the fibrin strands compared with the inner layers. Precipitation of the SNPs was patchy in the outer layers and quite homogeneous in the inner layers. Moreover, the leukocytes were denser in the outer layers than in the inner layers.

Physical properties

The tensile strength of SNP-impregnated PRF was greater than PRF.

Degradation time

The control PRF was degraded in a time-dependent manner and virtually completely digested at 6 days of incubation. In contrast, initially, PRF initially turned the HBSS cloudy (8 days), but thereafter, the SNP-impregnated PRF did not show appreciable degradation for at least 15 days of incubation.


  Discussion Top


In the present study, the tensile strength was significantly higher in the SNP-impregnated PRF membranes than the PRF group. The membrane's mechanical properties were improved as a result of adding SNPs to the PRF and its array in the fibrin matrix. The microscopic assessment of the samples showed that the SNPs were more densely mixed with the fibrin strands in the outer layers than the inner layers. Furthermore, their precipitation was patchy in the outer layer but quite homogenous in the inner layers. This may justify the improved mechanical properties of the PRF membrane impregnated by SNPs. Yet, further studies by an electron microscope are suggested to investigate more details.

It is generally thought that a barrier membrane should be preserved at the implantation site for 3–4 weeks to enhance periodontal tissue regeneration and integration.[13] Among absorbable membranes, those made of synthetic polymers such as polyglycolic acid and polylactic acid copolymer demonstrate a slow degradation rate (=12 months), whereas collagen-based membranes degrade faster and have been reported to remain stable for 16–38 weeks without significant degradation.[15] However, noncrosslinked collagen membranes lose their structural integrity in 7 days. Walker KA et al.'s in vivo animal implantation study demonstrated that the PRF could degrade as fast as noncrosslinked collagen-based membranes.[13]

Therefore, it is possible that increased crosslinking density among individual fibrin fibers within a PRF could prolong the preservation of the PRF at the implantation site and allow it to serve as a more clinically optimal GTR membrane.

On the other side, the most common postoperative complication of regenerative surgeries is the membrane exposure, in which case, the oral cavity microorganisms can be colonized on the membrane and jeopardize the success of treatment.[10] Certainly, it results in increased infection risk and poor bone repair. Thus, reinforcing the antimicrobial feature of membranes can contribute to the improvement of the treatment outcome. The SNPs are highly biocompatible and have favorable properties such as antimicrobial property.[11] Despite the several proposed theories, controversy exists regarding the mechanism of action of SNPs on microbes.[14] SNPs can adhere to the cell membrane of the bacteria and make it porous, which consequently changes the permeability of the cell membrane and causes cell death.[15] A number of studies claimed that the contact between the SNPs and bacteria forms free radicals which can damage the bacterial cell membrane and cause bacterial death through increasing the permeability.[10] It was also announced that the Ag ions released from the nanoparticles could interfere with the thiol group of enzymes and deactivate them.[14] Several studies showed that these materials can influence a wide spectrum of Gram-negative and Gram-positive bacteria and also antibiotic-resistant species. They have also been proven to have antifungal and antiviral effects.[10] Bone cement reinforced with nanosilver can destroy different bacteria such as Staphylococcus epidermidis, methicillin-resistant, in vitro. Adding SNPs to oral mouthwashes considerably reduced the growth of Streptococcus mutans compared with antibiotics and chlorhexidine.


  Conclusion Top


In the present study, SNP-impregnated PRF yielded a product which can help prevent the growth of a great family of bacteria viridans group streptococci on the surgical sites and its consequences. This membrane not only has biological advantages but also offers better mechanical properties, including higher tensile strength and degradation time compared with the traditional membrane.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Pellegrini G, Pagni G, Rasperini G. Surgical approaches based on biological objectives: GTR versus GBR techniques. Int J Dent 2013;2013:521-47.  Back to cited text no. 1
    
2.
Sculean A, Nikolidakis D, Schwarz F. Regeneration of periodontal tissues: Combinations of barrier membranes and grafting materials – Biological foundation and preclinical evidence: A systematic review. J Clin Periodontol 2008;35:106-16.  Back to cited text no. 2
    
3.
Kumar VR, Gangadharan G. Platelet rich fibrin in dentistry: A review of literature. Int J Med 2015;3:72-6.  Back to cited text no. 3
    
4.
Castro AB, Meschi N, Temmerman A, Pinto N, Lambrechts P, Teughels W, et al. Regenerative potential of leucocyte – And platelet-rich fibrin. Part A: Intra-bony defects, furcation defects and periodontal plastic surgery. A systematic review and meta-analysis. J Clin Periodontol 2017;44:67-82.  Back to cited text no. 4
    
5.
Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part II: Platelet-related biologic features. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006;101:e45-50.  Back to cited text no. 5
    
6.
Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: From pure platelet-rich plasma (P-PRP) to leucocyte-and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009;27:158-67.  Back to cited text no. 6
    
7.
Lee SB, Kwon JS, Lee YK, Kim KM, Kim KN. Bioactivity and mechanical properties of collagen composite membranes reinforced by chitosan and β-tricalcium phosphate. J Biomed Mater Res B Appl Biomater 2012;100:1935-42.  Back to cited text no. 7
    
8.
Marturello DM, McFadden MS, Bennett RA, Ragetly GR, Horn G. Knot security and tensile strength of suture materials. Vet Surg 2014;43:73-9.  Back to cited text no. 8
    
9.
Feinberg SE, Aghaloo TL, Cunningham LL Jr. Role of tissue engineering in oral and maxillofacial reconstruction: Findings of the 2005 AAOMS research summit. J Oral Maxillofac Surg 2005;63:1418-25.  Back to cited text no. 9
    
10.
Zhang J, Xu Q, Huang C, Mo A, Li J, Zuo Y. Biological properties of an anti-bacterial membrane for guided bone regeneration: An experimental study in rats. Clin Oral Implants Res 2010;21:321-7.  Back to cited text no. 10
    
11.
Ge L, Li Q, Wang M, Ouyang J, Li X, Xing MM. Nanosilver particles in medical applications: Synthesis, performance, and toxicity. Int J Nanomedicine 2014;9:2399-407.  Back to cited text no. 11
    
12.
Khorshidi H, Haddadi P, Raoofi S, Badiee P, Dehghani Nazhvani A. Does adding silver nanoparticles to leukocyte- and platelet-rich Fibrin improve its properties? Biomed Res Int 2018;2018:8515829.  Back to cited text no. 12
    
13.
Chaudhari D, et al. The Heat-compressed platelet-rich fibrin preparation as a barrier membrane. Int J Curr Adv Res 2018;7:11978-80.  Back to cited text no. 13
    
14.
Gunasekaran T, Nigusse T, Dhanaraju MD. Silver nanoparticles as real topical bullets for wound healing. J Am Coll Clin Wound Spec 2011;3:82-96.  Back to cited text no. 14
    
15.
Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: A case study on E. coli as a model for gram-negative bacteria. J Colloid Interface Sci 2004;275:177-82.  Back to cited text no. 15
    


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  [Figure 1], [Figure 2], [Figure 3]



 

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