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Review

Optimized centrifugation preparation of the platelet rich plasma:

Literature review
B. Croisé a,b,*, A. Paré a,b, A. Joly a, A. Louisy a, B. Laure a,b, D. Goga a,b
a
Department of Maxillofacial and Plastic Facial Surgery, Trousseau Hospital, CHU Trousseau, Avenvue de la république, 37170 Chambray-lès-Tours, France
b
School of medicine, University of François Rabelais, 370 Tours, France

A R T I C L E I N F O A B S T R A C T

Article history: Nowadays, the Platelet Rich Plasma (PRP) is frequently used for its therapeutic effects on wound healing,
Received 12 April 2019 and this due to secretion of many growth factors. However, no standardized procedure has been set up.
Accepted 2 July 2019 The aim of this article is to check the various preparations (centrifugation time and speed). This review
recorded all the international articles published between 2007 and 2018, for which the assessment
Keywords: criteria were the platelet concentration and/or the growth factor release rate. A multitude of protocols
Platelet Rich Plasma has been looked at with a simple or double centrifugation. All of them have shown an increase in the
Centrifugation
platelet concentration allowing a therapeutic effect. However, when the centrifugation force is extended,
Preparation
Platelet concentration
platelets can possibly be altered. The diversity of methods can be linked to the use of various centrifuges.
Maxillofacial surgery A procedure with simple centrifugation would be a good compromise for the day-to-day use of the PRP in
surgery.
C 2019 Elsevier Masson SAS. All rights reserved.

1. Introduction
The Platelet Rich Plasma (PRP) is a plasma rich in platelets,
obtained from venous blood.
In order to get a therapeutic aimed PRP, the concentration must
be between 200  103 and 1000  103 platelets/mL. A higher
concentration would be unfavourable [1].
Many studies showed that, during contact with collagen, PRP
released growth factors such as Platelet Derived Growth Factor
(PDGF), Vascular Endothelial Growth Factor (VEGF), Epidermal
Growth Factor (EGF), Platelet Factor 4 (PF-4), Insulin like Growth
Factor-1 (IGF-1) and, Transforming Growth Factor Beta (TGF b) by
their a-granules [2].
These growth factors play a key role in the early stage of wound
healing by allowing a stem cell proliferation and, angiogenesis.
Many medical specialities such as orthopedic, ophthalmic and
maxillofacial surgery are looking for this tissue engineering effect.
In our day-to-day practice in maxillo facial surgery, we use PRP
in preimplant surgery during bone grafts, in order to get a good
* Corresponding author at: Department of Maxillofacial and Plastic Facial
Surgery, Trousseau Hospital, CHU Trousseau, Avenvue de la république,
37170 Chambray-lès-Tours, France.
E-mail address: benjamin.croise.med@gmail.com (B. Croisé).
mucosal healing: an important and essential step to avoid the main
complication linked to this surgery: infection. Indeed, a bad
mucosal healing is a source for higher infection probability, due to
the fact that the graft is exposed in a so-called dirty environment
[3]. Moreover, PRP use allows a quicker haemostasis and a better
bone regeneration [4].
However, in spite of PRP substantial use in our specialities, no
centrifugation protocol based on time, speed and centrifugation
numbers is defined to obtain a good quality PRP, leading to an
optimal effect on healing.
It was a topical issue, since the first articles on the question
dating back to the seventies disagreed.
Slitcher et al. [5] drew up a double centrifugation protocol with
a first centrifugation at 1000 G during 9 min (min), then 3000 G
during 20 min, while Kahn et al. [6], during the same year, set up a
protocol with only one centrifugation at 3800 RPM during 4 min.
Therefore, many questions arise concerning the PRP prepara-
tion: Is only one centrifugation enough to obtain a clinical effect on
the healing? What are centrifugation time and speed to obtain a
good quality PRP?
The aim of this article is to draw up a literature review to define
a simple and reproducible protocol (centrifugation optimal time
and speed) in PRP preparation to get an ideal platelet yield, whilst
avoiding its alteration.

https://doi.org/10.1016/j.jormas.2019.07.001
2468-7855/ C 2019 Elsevier Masson SAS. All rights reserved.

Please cite this article in press as: Croisé B, et al. Optimized centrifugation preparation of the platelet rich plasma: Literature review. J
Stomatol Oral Maxillofac Surg (2019), https://doi.org/10.1016/j.jormas.2019.07.001
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2. Materials and Methods PRP preparation:

2.1. 1Selection  whole, non-anticoagulated blood collection in a citrated tube;

This systematic literature review looked into all international
articles published between 2007 and 2018 on Medline (PubMed).
The used key words combination was ‘‘Platelet Rich Plasma’’ and
‘‘centrifugation’’ and ‘‘platelet concentration’’. All the titles and
abstracts of the identified articles on the database have been
analyzed in order to evaluate their eligibility. The review should
study a precise protocol of centrifugation to obtain a platelet
concentration. The sought judging criteria are the platelet yield
and/or the release rate of growth factors. The articles on studies
carried out on animals have been excluded from the study, as well
as articles dealing with other protocols than centrifugation.
Moreover only the English and French articles have been analyzed.
Among the 95 articles in the database searching, 21 articles were
eligible and after full-text analysis, 14 articles were included in this
systematic review (Fig. 1).
 first centrifugation (soft spin) in order to separate the red blood
cells (lower layer 45%) from the rich in platelets plasma (upper
layer 55%). There is a middle leucocyte layer or buffy coat (<1%);
 sample of the upper layer;
 then, there are two possible choices:
 either we use this rich in platelets plasma,
 or we carry out a second centrifugation (hard spin) with a
higher speed to obtain a platelet concentrated plasma or pure
PRP, made of granules, which is represented by the lower
layer. This coat is homogenized with plasma.
2.2. Various centrifuges
The most used centrifuges during these studies are presented in
this chapter:

Fig. 1. Systematic review study selection process: flow diagram of articles identified and excluded for the study.

Please cite this article in press as: Croisé B, et al. Optimized centrifugation preparation of the platelet rich plasma: Literature review. J
Stomatol Oral Maxillofac Surg (2019), https://doi.org/10.1016/j.jormas.2019.07.001
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 Eppendorf 5810 R [4] with a maximum speed of 14,000 RPM;
 Rotina 380 R [7] with a maximum speed of 15,000 RPM;
 Jouan BR4i [8] with a maximum speed of 14,000 RPM;
 Allegra X15R [9] with a maximum speed of 10,200 RPM;
 Kubota 5900 [2] with a maximum speed of 8500 RPM.
Formulae
In our opinion, a formula must be defined, due to the language
difference noticed in these articles. Indeed, some authors draw up
their protocols by using the Gay (G) measure unit, while others use
the rotation/min unit (RPM) to speak of the centrifugation speed;
G = 11,18  10 6  r (cm)  N2 5RPM) in which r is the rotor
rotation radius
N= (G/(1.118  10 5  r))
Results
It has been decided that it is easier to use a chronological order
in this literature review to allow the study of practice develop-
ments.
Indeed, Tamimi et al. [10] in 2007 showed in his study that a
double centrifugation enabled to obtain a PRP with higher platelet
concentrations than during a simple centrifugation. However, the
double centrifugation system caused modifications in the platelet
ultrastructure. He drew up a protocol of 160 G for 10 min, then
400 G for 10 min.
During the same year, Rutkowski et al. [11] carried out a study
to obtain a simple and reproducible method for PRP preparation.
He based it on 7 subjects and his judging criteria were the platelet
concentration and whether the utilization time plays a lead role on
its growth factors release. The set up protocol for a good quality
PRP without platelet alteration was 1350 G for 10 min. His study
also showed that a longer time (> 3 h) caused a decrease of TGF b
concentration but without alteration of the other growth factors.
Mazzocca et al. [1] set up a study with 8 healthy subjects to
identify the best protocol among three standards, a low speed
centrifugation (1500 RPM during 5 min), a high speed centrifuga-
tion (3200 RPM during 15 min) and a double centrifugation with a
soft pin of 1500 RPM during 5 min and a hard spin of 6300 RPM
during 20 min. A significant difference has been noted for the high
speed centrifugation with a platelet concentration of
873,8  207,2  103/mL (P < 0,05) compared with the low speed or
the double centrifugation. This centrifugation protocol significantly
increases the growth factors production with a sole exception, the
VEGF-A production. Then, Mazzocca [1] concluded that the appro-
priate protocol for a PRP production with respect to the required
clinical effect is 3200 RPM for 15 min.
Araki et al. [2] studied several double centrifugation protocols. At
first, he varied different speeds from 30 to 2330 G during 10 min to
carry out the « soft » spin. He obtained the best platelet concentration
(3 times higher than whole blood concentration) with a protocol
from 230 to 270 G for 10 min. He noticed that platelet concentration
decreased after 320 G and for a speed higher than 840 G, the
removed blood cells precipitated. The obtained protocol for the
« hard » spin was 2330 G for 10 min, which allowed a platelet
concentration 7.4 times higher than the PRP with a one tenth PCP
(Platelet Concentrated Plasma) reduced volume, i.e. 20 times more
than in whole blood. He also noticed that the addition of
Ethylenediaminetetraacetic anticoagulant (EDTA) allowed to avoid
the platelet aggregation, then a better platelet concentration.
In 2012, Bausset et al. [12] set up a double concentration
protocol to obtain a PRP with the best platelet concentration. He
carried out his study with 54 healthy subjects. The first
Please cite this article in press as: Croisé B, et al. Optimized centrifugation preparation of the platelet rich plasma: Literature review. J
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3
centrifugation at 130 G for 15 min was decided in advance. Then,
he studied several speeds 130, 250, 400 and 1000 G for the second
step. The speed allowing to obtain the best platelet concentration
without platelet aggregation was 250 G for 15 min with a
significant difference (P = 0.02). The set up protocol consisted of
two serial 15-minute centrifugations of anticoagulated whole
blood at 130 G for the « soft » spin and, 250 G for the « hard » spin.
Ogundipe et al. [13] studied a double centrifugation with
30 healthy volunteers. The first centrifugation was 1000 RPM for
10 min and the second one 1200 RPM for 10 min. The whole  -
platelet concentration averaged 268,667 platelets/mL3. After this
double centrifugation, it increased to 3,157,667 platelets/mL3,
11.8 times more and, a growth factor enrichment of 1.086%.
In his study, Franco et al. [14] carried out a first centrifugation at
400 G for 10 min. Then, he centrifuged the buffy coat at 800 G for
10 min and reduced the volume by two-thirds to obtain the PRP.
This protocol resulted in a platelet concentration 8.5 times higher
(6.03  108 platelets/mL).
Jo et al. [9] also studied the best protocol with 39 samples of
healthy subjects. The platelet concentration of these samples was
152.8  103  28.9 mL. A method of double centrifugation was
carried out. The first centrifugation was evaluated from 500 G to
1900 G for 5 min and, from 100 G to 1300 G for 10 min. Then, the most
efficient centrifugation was centrifuged a second time at different
protocols (1000 G and, 1500 G for 15 min and, 10 min, 2000 G for
5 min or 3000 G for 5 min).
The first centrifugation allowing a high platelet concentration is
900 G for 5 min, namely an increase of 92% for a platelet
concentration of 310.7  103  78.5 mL. The second centrifugation is
then carried out on these samples and the obtained protocol is 1500 G
for 15 min, allowing to get a platelet concentration of
633.2  91.6  103/mL, namely 4.2 times more.
Thus, the Jo protocol consisted in a double centrifugation,
namely 900 G for 5 min, then 1500 G for 15 min.
In his study, Amable et al. [8] wished to set up a repeatable and
optimal method for PRP preparation. Therefore, he checked up a
double centrifugation PRP preparation on 22 healthy subjects. The
best result for platelet concentration was 300 G for 5 min in the
first centrifugation. The second centrifugation allowing a lower
platelet loss was 700 G for 17 min.
In the Perez survey [7], the best double centrifugation protocol
has been searched. The first centrifugation was made from 50 to
820 G for 10 min. After this first centrifugation, the upper coat has
been taken out and centrifuged at various speeds (200, 400, 800,
1200 and 1600 G) for 10 min.
The first centrifugation speed selected was 100 G for 10 min;
the selected speed for the second centrifugation was 400G for
10 min. This protocol, after removal of two thirds of plasma after
the second centrifugation allowed to get a 5 times higher platelet
concentration and to keep the platelet vitality.
Sabarish et al. [4] studied 3 double centrifugation methods with
20 healthy subjects. The first protocol was set up by Marx in 1999,
namely a first centrifugation at 1000 RPM for 4 min and, a second
one at 800 RPM for 9 min. The second method by comparison was
the Okuda one in 2003, with 2400 RPM for 10 min, then 3600 RPM
for 15 min. The last method was the Landesberg one in 2000 with
1400 RPM for 10 min, then 1600 RPM for 10 min.
The best results to have the highest platelet concentration were
obtained with 1000 RPM for 4 min and, then 800 RPM for 9 min.
This method allows an increase of 1,6 times (2,18  108 to
3,5  108).
In his article, Arora et al. [15] wanted to study the platelet
concentration and the release rate of the various growth factors
through different double centrifugation protocols. He set up
9 groups for the first centrifugation in terms of force (A: 100 G, B:
208 G, C: 440 G) and time (1: 5 min, 2: 10 min, 3: 20 min).
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The highest platelet concentration was found in the C3 group.
He also noticed that, when the centrifugation time increased, the
platelet concentration became higher. Likewise, when the centri-
fugation force increased at 10 min, the platelet concentration also
increased, but this causal link was not found, when the
centrifugation time was 20 min.
In order to study the growth factor release, the various tubes
were centrifuged at 172 G for 3 min and, after a night of incubation,
at 1552 G for 23 min. The study highlighted that there was no
significant difference in the PDGF-AB release. The TGF B secretion
significantly increased in all the groups, except when time was
20 min. The VEGF release increased in all the groups, without any
significant difference between them.
Arora et al. [15] concluded that the best protocol was 208 G for
20 min, because the increasing centrifugation force leaded to a
platelet aggregation and a bad release of growth factors.
The aim of Eren’s [16] article was to study the centrifugation
time effect on the growth factors release. He checked the platelet
concentration and the growth factors rate at different rest times on
20 healthy subjects (1 h, 24 h and 72 h), using 2 protocols, one at
2660 RPM for 10 min (Group A) or 12 min (Group B). The
referenced platelet concentration was 235,3  52,3/mL  103, the
result for the group A was 218,7  50,1/mL  103 and for the group B
214,7  52,1/mL  103: there is no significant difference.
As the growth factor rate, the VEGF rate was significantly higher
after centrifugation during 12 min.
This study showed that centrifugation time had no effect on
platelet concentration and growth factors other than VEGF.
However VEGF being very important in the tissue healing process,
it would be preferable to increase the centrifugation time.
Yin et al. [17] in this study wanted to find an optimal of a double
centrifugation in 80 volunteers patients.
For the first centrifugation he studied 6 different protocols
(110 G for 15 mn (110  15), 130  10, 130  15, 160  10,
160  15, 180  10).
The second centrifugation was also studied by 6 various
protocols (180  10, 180  15, 250  10, 250  15, 450  10,
450  15).
The platelet concentration of PCP obtained using 160  10 was
significantly higher than that of PCP obtained using 110  15
(P < 0.001), 130  10 (P < 0.001), 160  15 (P < 0.001) and
180  10 (P < 0.001), and comparable with that obtained using
130  15 (P > 0.999). The result was similar for leukocytes and
growth factor concentration. Therefore, 160  10 was designated
as the optimal centrifugation conditions for the first spin.
The second spin was realized in the sample where the first spin
was 160  10. The best result was obtained by the protocol
250  15. In fact, the platelet enrichment factor of P-PRP obtained
using 250  15 was significantly higher than that obtained using
180  10 (P < 0.001), 180  15 (P < 0.001) and 250  10
(P < 0.001). However, the platelet enrichment factor did not
further increase by preparation using 450  10 (P > 0.999) and
450  15 (P > 0.999) compared with 250  15. But
250  15 allowed to have a concentration of growth factor more
important. P-PRP obtained by centrifugation at 250  15 had a
higher level of PDGF-AB compared to that obtained at 180  10
(P = 0.001), 180  15 (P = 0.028), 250  10 (P = 0.008), 450  10
(P = 0.011) and 450  15 (P = 0.004) (Table 1).
3. Discussion
Many protocols were studied, allowing an increase of the PRP
platelet concentration. However, it is important to distinguish the
methods of simple and double centrifugation; the second one
allowing to get a platelets concentrated plasma with a longer
preparation.
The platelet concentration allowing a therapeutic effect is
obtained in all the simple centrifugation protocols.
Moreover these articles also show that the platelets are fragile
cells and whether the centrifugation force is too strong, there is a
risk of alteration and then inefficiency. However, the VEGF
concentration increases according to the centrifugation time.
A method of simple centrifugation would then be a good
compromise.
In our day-to-day practice, we use a Sigma centrifuge and a
protocol of simple centrifugation at 3000 RPM (or 1350 G) for
3 min.
A new method described by Wu [18] would allow a more
standardized protocol, with a shorter preparation time. The use of
ultrasounds on the whole blood would result in a concentration
comparable to that of centrifugation.
PRP indication in maxillofacial surgery is also contested. Indeed,
Betega et al. [19] shows in his article that the PRP clinical benefit is
not proven. However, in 2013, Albanese et al. [20] studied PRP use
in several maxillofacial indications. This review points out that PRP
use after a tooth extraction allows to improve healing but does not
contribute to bone regeneration. As to indication in periodontal
surgery, the results are heterogeneous. The review also shows
satisfactory results in pre-implant and implant surgery, depending
on the type of material being used. This interesting outcome is also
noted in osteonecrosis mandibular surgery. Fornaini et al. [21]
states a case with PRP treatment in osteonecrosis mandibular
surgery.

Table 1
Summary table of various protocols.

Study/Year 1ST centrifugation 2ND centrifugation Platelet concentration

Rutkowski/2007 1350 G for 10 min 6 times more concentrated

Tamimi/2007 160 G for 10 min 400 G for 10 min 630,2  103/mL
Mazzocca/2012 3200 RPM for 15 min 873,8  207,2  103/mL
Araki/2012 230 to 270 G for 10 min 2330 G for 10 min 7,4 higher
Bausset/2012 130 G for 15 min 250 G for 15 min 3,96 times higher
Ogundipe/2012 1000 RPM for 10 min 1200 RPM for 10 min 11,8 times higher
Franco/2012 400 G for 10 min 800 G for 10 min 8,5 times higher
Jo/2013 900 G for 5 min 1500 G for 15 min 633,2  91,6  103/mL
4,2 times higher
Amable/2013 300G for 5 min 700 G for 17 min 140 to 190  103/mL
Perez/2014 100 G for 10 min 400 G for 10 min 5 times higher
Sabarish/2015 1000 RPM for 4 min 800 RPM for 9 min 3,5  108
1,6 times higher
Arora/2016 208 G for 20 min 1552 G for 23 min 87% relative platelet concentration
Eren/2016 2660 RPM for 12 min 214,7  52,1/mL  103
Yin/2017 160 G for 10 min 250 G for 15 min More 1250  109/L

Please cite this article in press as: Croisé B, et al. Optimized centrifugation preparation of the platelet rich plasma: Literature review. J
Stomatol Oral Maxillofac Surg (2019), https://doi.org/10.1016/j.jormas.2019.07.001
G Model
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Picard et al. [22] conducted a literature review showing PRP
benefit in alopecia treatment.
Hersant et al. [23] goes even further in using PRP in free flap.
The PRP use area remains contested, and neither protocols nor
indications have been set up but PRP can still be used in a lot of
maxillofacial indications.
4. Conclusion
This literature review shows the complexity of implementing a
standardized protocol for the PRP preparation. The use of various
centrifuges and units of measure (Gay or RPM) is a drag for results
repeatability. It is then necessary to convert the units to fit to the
used centrifuge.
Disclosure of interest
The authors declare that they have no competing interest.
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