PRP as a New Approach to Prevent Infection: Preparation and In vitro Antimicrobial Properties of PRP

Summary

Implant-associated infection is a significant clinical complication. This study describes an approach using platelet-rich plasma (PRP) to prevent implant-associated infections, presents the protocol for preparing PRP with constant platelet concentration, and reports the newly identified antimicrobial properties of PRP and related protocols for examining such antimicrobial properties in vitro.

Abstract

Implant-associated infection is becoming more and more challenging to the healthcare industry worldwide due to increasing antibiotic resistance, transmission of antibiotic resistant bacteria between animals and humans, and the high cost of treating infections.

In this study, we disclose a new strategy that may be effective in preventing implant-associated infection based on the potential antimicrobial properties of platelet-rich plasma (PRP). Due to its well-studied properties for promoting healing, PRP (a biological product) has been increasingly used for clinical applications including orthopaedic surgeries, periodontal and oral surgeries, maxillofacial surgeries, plastic surgeries, sports medicine, etc.

PRP could be an advanced alternative to conventional antibiotic treatments in preventing implant-associated infections. The use of PRP may be advantageous compared to conventional antibiotic treatments since PRP is less likely to induce antibiotic resistance and PRP's antimicrobial and healing-promoting properties may have a synergistic effect on infection prevention. It is well known that pathogens and human cells are racing for implant surfaces, and PRP's properties of promoting healing could improve human cell attachment thereby reducing the odds for infection. In addition, PRP is inherently biocompatible, and safe and free from the risk of transmissible diseases.

For our study, we have selected several clinical bacterial strains that are commonly found in orthopaedic infections and examined whether PRP has in vitro antimicrobial properties against these bacteria. We have prepared PRP using a twice centrifugation approach which allows the same platelet concentration to be obtained for all samples. We have achieved consistent antimicrobial findings and found that PRP has strong in vitro antimicrobial properties against bacteria like methicillin-sensitive and methicillin-resistant Staphylococcus aureus, Group A Streptococcus, and Neisseria gonorrhoeae. Therefore, the use of PRP may have the potential to prevent infection and to reduce the need for costly post-operative treatment of implant-associated infections.

Introduction

Implant-associated infection is a significant clinical complication. Staphylococcus aureus (S. aureus) is one of the most common microorganisms isolated from implant-associated infections. It is capable of producing a biofilm that covers the surfaces of implants and may lead to antibiotic-resistant infection ^1,2. Treatment of implant-associated infection frequently requires long-term hospitalization for repeated debridements and prolonged parenteral antibiotic therapy. In antibiotic resistant cases, removal of the implant may be necessary. The rising resistance of bacteria to antibiotics has also been referred to by the Centers for Disease....

Protocol

1. Preparation and Activation of PRP

1.1 Blood draw

1. Anesthetize rabbit by inhalation of isoflurane (2% in O₂ for induction and 1% for maintenance).
2. Draw 2 ml 0.129 M tri-sodium citrate (an anticoagulent solution) into a 20 ml syringe. The tri-sodium citrate solution is prepared by dissolving 1.897 g tri-sodium citrate in 50 ml distilled H₂O and filtering with a 0.22 μm sterile filter.
3. Sterilize the rabbit ear using 70% ethanol.
4. Draw blood (e.g. 5 ml) from the rabbit ear vein via a butterfly needle (25 G) connected to the syringe.
5. Mix the blood with the tri-so....

Results

PRP is reproducibly prepared using a twice centrifugation approach (Figure 1). PRP is found to present strong (up to 100-fold reduction in CFUs) in vitro antimicrobial properties against methicillin resistant S. aureus (MRSA) (Figure 3), which is commonly found in hospitals worldwide ¹⁴. Similarly, PRP has strong antimicrobial properties against methicillin sensitive S. aureus (MSSA), Group A Streptococcus, and Neisseria gonorrhoeae.......

Discussion

Platelet-rich plasma has been increasingly used for clinical applications due to its healing-promoting properties ^15-17. In the present study, PRP was presented as a new approach for infection prevention. PRP was found to have strong antimicrobial properties against MRSA, MSSA, Group A Streptococcus, and Neisseria gonorrhoeae. The major advantages of PRP, compared to conventional antibiotic treatments, for infection prevention include: (1) Current antibiotic therapies are facing challenges in.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Bovine thrombin	King Pharmaceuticals, Inc	60793-215-05	Thrombin (bovine origin)
Calcium chloride	King Pharmaceuticals, Inc	60793-215-05	10% calcium chloride
Ethanol	Sigma-Aldrich	E7023
Isoflurane	Baxter	1001936060
Mueller Hinton broth	Becton, Dickinson and Company	275710
Phosphate-buffered saline	Sigma-Aldrich	D8662
Tri-sodium citrate	Sigma-Aldrich	W302600
Tryptic soy agar	Fisher Scientific	R01202
Centrifuge	Kendro Laboratory Products	750043077
Syringe filter	Millipore	SLGP033RS