2013 pelletier prp constituents

74 Clinical Sciences

Pelletier MH et al. Platelet Function and Constituents … Int J Sports Med 2013; 34: 74–80

accepted after revision

May 10 , 2012

Bibliography

DOI http://dx.doi.org/

10.1055/s-0032-1316319

Published online:

August 14, 2012

Int J Sports Med 2013; 34:

74–80 © Georg Thieme

Verlag KG Stuttgart · New York

ISSN 0172-4622

Correspondence

Dr. Matthew H Pelletier, PhD

Surgical & Orthopaedic

Research Labs

University of New South Wales

Level 1

Clinical Sciences Bld

2031 Randwick

Australia

Tel.: +61/293/822 687

Fax: +61/293/822 660

[email protected]

Key words

● ▶ platelet rich plasma

● ▶ blood

● ▶ autologous

● ▶ therapy

● ▶ p-selectin

● ▶ viable

Platelet Function and Constituents of Platelet Rich Plasma

dermal growth factor (EGF). These secreted pro-

teins have important roles in healing and tissue

regeneration [ 1 , 5 , 27 , 40 ] . Although there are

many proteins that may promote wound healing

in PRP, such as fi brinogen, the basic premise for

PRP remains the delivery of a soup of platelet-

derived proteins at concentrations above base-

line [ 13 – 15 , 50 ] .

PRP can be generated by centrifugation of antico-

agulated whole blood and collected with pipette

or syringe, but interest in PRP therapies has led to

the development of several commercial systems

aimed at improving ergonomics, decreasing vari-

ability, and improving platelet recovery in a

closed system. While these devices do produce

PRP with a concentration of platelets above cir-

culating baseline blood levels [ 29 ] , the product

varies between devices, leading Ehrenfest et al.

[ 11 ] to propose a classifi cation to highlight their

properties. Specifi cally, the classifi cation takes

account of diff erences in production including

whether or not the buff y coat (which contains a

near 7-fold increase in leukocytes [ 7 ] ) is col-

lected, fi brin and fi brinogen characteristics,

collection effi ciency, as well as practical consid-

Introduction

▼ Clinical use of Platelet Rich Plasma (PRP) has

been reported since the early 1990s for oral and

maxillofacial surgery as a source of growth fac-

tors to increase the rate and degree of bone for-

mation at early time points [ 31 ] . Since then PRP

has found an increasing use in a variety of clinical

applications. Primarily PRP has been used to sup-

port hard and soft tissue healing and formation

[ 5 , 9 , 35 , 44 ] , but has also extended to other uses

such as on nervous tissue [ 10 , 42 ] , in facial plastic

and cosmetic surgery [ 6 , 28 , 39 ] , burns [ 36 ] and

chronic leg ulcers [ 46 ] . The autologous nature of

PRP also alleviates concerns of transmissible dis-

eases or immunogenic reactions [ 34 ] .

The potential clinical benefi t of PRP relies on the

maintenance of intact platelet -granules. Upon

activation, platelets degranulate and secrete an

array of mitogenic and chemotactic growth factors, which include platelet derived growth

factor (PDGF- , PDGF- , PDGF- ), transform-

ing growth factor (TGF- 1, TGF- 2), vascular

endothelial growth factor (VEGF), fi broblast

growth factor (FGF), interleukin-1 (IL-1), and epi-

Authors M. H. Pelletier 1 , A. Malhotra 1 , T. Brighton 2 , W. R. Walsh 1 , R. Lindeman 2

Affi liations 1 Surgical & Orthopaedic Research Labs, University of New South Wales, Randwick, Australia 2 Department of Haematology, Prince of Wales Hospital, Randwick, Australia

Abstract

▼ Platelet Rich Plasma (PRP) therapies require

blood to be processed prior to application, how-

ever, the full assessment of the output of platelet

sequestration devices is lacking. In this study the

products of the Autologous Fluid Concentrator

(Circle Biologics TM , Minneapolis, MN) and the

Gravitational Platelet Separation System (GPS,

Biomet, Warsaw, IN, USA) were evaluated in

terms of platelet viability and PRP constituents.

The AFC and GPS produced 6.4 ( ± 1.0) ml and

6.3 ( ± 0.4) ml of PRP, with platelet recovery of

46.4 % ( ± 14.7 %) and 59.8 % ( ± 24.2 %) produc-

ing fold increases of platelets of 4.19 ( ± 1.62)

and 5.19 ( ± 1.62), respectively. Fibrinogen con-

centration was increased above baseline PPP

produced with the AFC. pH was lower for both

of the processed samples than for whole blood.

White Blood Cell count was increased around 5

fold. Functional tests showed preserved viabil-

ity with both devices. This represents essential

knowledge that every treating physician should

have before they can confi dently administer PRP

therapy produced by any method. These are the

fi rst published results of platelet function for the

GPS system and the fi rst performance results of

the AFC system. The PRP produced is classifi ed

according to broad classifi cations as Leukocyte-

PRP (L-PRP) for both devices.

75Clinical Sciences


erations such as size, weight of the centrifuge, and ease of use.

The development of this classifi cation will hopefully resolve

variable and sometimes confl icting results regarding PRP thera-

pies in the literature [ 19 , 28 , 30 , 37 , 38 , 44 , 47 ] .

Here we present a fi rst evaluation of the Autologous Fluid Con-

centrator (AFC) (Circle Biologics TM , Minneapolis, MN), a platelet

sequestration device which has recently been granted approval

by the Food and Drug Administration for use in the United States,

and a paired comparison of an established device (GPS, Biomet,

Warsaw, IN, USA).

Materials and Methods

▼ Both devices yield PRP, with the GPS system additionally produc-

ing Platelet Poor Plasma (PPP) and the AFC off ering the option of

PPP or Concentrated Platelet Poor Plasma (c-PPP). An evaluation of

c-PPP, PRP, and whole blood included blood count, platelet count,

pH, white cell count, platelet recovery, and fi brinogen concentra-

tion. Platelet function testing on obtained PRP was performed in a

smaller subset of samples at 0 and 4 h to investigate platelet reac-

tivity and short-term stability. These tests included assessments

of p-selectin levels while resting and upon activation with Adeno-

sine Diphosphate (ADP), reaction to hypotonic shock and agonist-

induced light transmission aggregometry.

Subjects This study conforms to ethical standards in sport and exercise

research as described by Harriss, et al. [ 21 ] . Volunteers were

recruited at the University of New South Wales and Prince of

Wales Hospital following approval of the Human Research Ethics

Committee. A total of 64 subjects were recruited for the study.

Volunteers were excluded if taking anticoagulant, anti-platelet,

or anti-infl ammatory medication, or aspirin. Subjects were de-

identifi ed by assigning a number relating to all subsequent blood

test results.

Blood collection Blood was collected from the medial cubital vein with a 19 G

needle by trained phlebotomists. Blood was collected in a 4 ml

EDTA Vacutainer for Full Blood Count (FBC), a 4.5 ml Li-Heparin

Vacutainer for standard biochemistry, a 3.5 ml Citrate Vacutainer

for fi brinogen concentration, 1 ml heparin syringe for pH, and for

each device 2 × 30 ml syringes preloaded with acid-citrate-dex-

trose formula A (ACD-A) anticoagulant, 10 % ( ● ▶ Fig. 1 ).

Platelet sequestration The centrifuge and AFC devices arrived in sterile packages ready

for use. The AFC is a 2 chamber modular system. The devices

tested in this study utilised a plasma concentration chamber at

the rear and a chamber that, when viewed from the front, resem-

bled an hourglass. Inside the hourglass chamber there is a col-

lection tube with a moveable collection area. The GPS system

uses a buoy of specifi c density within a cylindrical chamber

( ● ▶ Fig. 2 ).

Blood and ACD-A fi lled syringes were emptied into the devices

which were centrifuged at 3 200RPM for 15 min at room tem-

perature. Following centrifugation the plasma, red cell and buff y

coat layers were clearly evident ( ● ▶ Fig. 3 ).

PPP was drawn off with syringes connected to respective ports

and was further processed by 3 fi lter passes for the AFC resulting

in (c-PPP). PRP was drawn off with a moveable collection win-

dow between 2 O-ring seals with the AFC and via a separate port

with the GPS device.

For the collection of PRP, the selector valve was turned back to

“draw”, the collection window was moved so that the top seal

was level with the top of the buff y coat and all available fl uid was

drawn off using a 10 ml syringe connected to port B. A portion of

both PRP, PPP and, where processed, c-PPP was placed in capped

test tubes for FBC (n = 59) and fi brinogen concentration (n = 59)

and a Li Heparin syringe for pH (n = 59). A portion of PRP was

also placed in a capped tube for tests of platelet function (n = 12

for each test).

Blood assays All blood assays were performed at the South Eastern Area Labo-

ratory Services (SEALS) which is accredited by the National

Association of Testing Authorities (NATA), Australia’s govern-

ment-endorsed, international laboratory accreditation body.

59 matched samples of whole blood, PRP and PPP were evalu-

ated for pH, fi brinogen concentration and FBC. pH was measured

on an ABL800 FLEX Radiometer Blood Gas Analyser (Radiometer,

Copenhagen). Full Blood Count was performed on a Roche Sys-

mex Model XE-2 100 (Sysmex Corp., Kobe, Japan). Biochemistry

assays were performed on a Beckman Coulter UniCel DXC880

blood analyzer (Beckman Coulter, USA). Fibrinogen assays were

performed on an STA R Evolution Coagulation Analyzer (Stago,

USA). Platelet recovery was calculated from platelet counts as

100 % *(platelet count of PRP * volume of collected)/(Platelet

count of whole blood * volume of whole blood). Platelet concen-

tration factor was calculated as Platelet count of PRP/whole

blood platelet count.

Surface P-selectin expression (CD62p antigen), as a measure of

platelet activation, was assessed by fl ow cytometry in resting

and activated state samples. Approximately 1 ml of PRP from

each device was set aside for p-selectin assays. Phosphate buff -ered saline (2.92 μl) or ADP agonist (40 μM) was added to 70 ml

subsamples of PRP. These samples were incubated for 15 min at

Fig. 1 Allocation of collected blood. Whole Blood in 2 x30ml syringes loaded

w (ACD-A) 10% foreach device

1 ml: Whole Blood(Heparin syringe)

4 ml: Whole Blood(EDTA vacutainer)

3.5 ml: Whole Blood(Citrate vacutainer)

4.5 ml: Whole Blood(Li-Heparin vacutainer)

AFC or GPSProcessing pH

PRP

Full Blood Count Biochemistry Fibrinogen

PPS; GPSc-PPP; AFC

pHFibrinogen

Full Blood Count

0h p-selectin resting0h p-selectin ADP0h Hypotonic Stress0h Aggregation




pH

Fibrinogen

Full Blood Count



37 °C. 4 × 5 μl aliquots were taken and incubated with CD41a-

PerCP antibody and either 1μl CD62-PE or 1 μl mouse IgG1-PE

(isotope control) (DB Biosciences, San Jose, CA). These samples

were incubated in the dark at room temperature for 20 min.

600 ml of Ringer’s solution was added following incubation.

Samples were analyzed in a BD FACS Canto II fl ow cytometer

(Becton, Dickinson and Company, USA). For determination of %

p-selectin expression, forward and light scatter and fl uorescence

were acquired using logarithmic scale. A total of 20 000 platelet

events were gated. The isotope matched control antibody was

used to set threshold for CD62P positivity.

Hypotonic shock response (HSR) measures the platelet’s ability

to recover its normal volume after swelling when exposed to a

hypotonic environment. HSR is an optical method and PRP

required an additional soft spin (900RPM/10 min) to remove

intervening red cells for output from the AFC device. The result-

ing PRP was removed from the red cells with a pipette and plate-

let concentrations adjusted to < 500 × 10 9 /ml and allowed to rest

Fig. 2 Left to Right; Circle Biologics and Biomet

devices, before use (top row) and after centrifuga-

tion (bottom row).

Fig. 3 Close up of buff y coat interface, Circle

Biologics (Left) and Biomet (Right). Black arrows

indicate the PPP portion, the White arrows indicate

the buff y coat, and the double white arrows indi-

cate the red cell pack.

77Clinical Sciences


for 30 min. 6 × 150 μl of PRP from each device was transferred

into the wells of a microtitre plate. 3 × 150 μl of saline was added

to PRP samples and the microtitre plate was placed in a BioTeck

EL808 Ultra Microplate Reader (BioTek, Instruments, Inc.,

Winooski, VT). 150 μl of H 2 O were added with a micropipette to

the remaining 3 samples simultaneously, the lid was closed and

reading commenced. Readings of light transmission was per-

formed at 405 nm for 10 min at 15 s intervals. Results were

recorded with this protocol via Gen5 Data Analysis Software

(Generation 5, Toronto, Canada). Light transmission (T) was con-

verted to optical density according to OD = − log 10 T or T = lO − OD .

The optical density (OD) of the saline samples acted as the base-

line for measurements. The diff erence in the peak OD and the

baseline divided by the diff erence in OD at 10 min and baseline

produced a percent recovery.

Aggregation was performed following preparation to remove

red cells as above. 240 μl samples of PRP, PPP and c-PPP were

tested in a 4 station AggRAM aggregometer (Helena Laborato-

ries, Gateshead, UK). All samples were stirred with magnetic stir

bars at 600 rpm. Following a quality check, the OD was meas-

ured in the matched samples of PPP, which acted as a control.

The PRP was then placed in the aggregometer. Continuous read-

ing of OD continued for 10 min while 10 ul of collagen aggregant

was added to achieve a fi nal concentration of 20 μg/ml and fi nal

volume of 250 μl. Results of all tests were compared with paired

t-tests using Tukey’s criteria and a signifi cance level of 0.05.

Results

▼ Of the 64 blood samples collected for the study, none were

excluded based on irregular biochemistry results, one was lost

to human error, and post processing assays were precluded by

insuffi cient volume in 4 samples resulting in a sample size of

n = 59 for fi brinogen, cell count and pH tests and n = 12 for all

platelet function assays. 54.1 ( ± 0.2) ml (AFC) and 55.7 ( ± 1.5) ml

(GPS) of whole blood plus ACD-A was loaded into the device and

6.4 ( ± 1.0) ml for the AFC and 6.3 ( ± 0.4) ml for the GPS of PRP

was collected. Platelet recovery from whole blood samples for

the AFC and GPS was 46.4 % ( ± 14.7 %) and 59.8 % ( ± 24.2 %),

respectively, representing concentration factors (fold increase)

of 4.19 ( ± 1.62) and 5.19 ( ± 1.62). Fibrinogen concentration was

increased above baseline for c-PPP (AFC only) but not PRP. pH

was lower for both of the processed samples than for whole

blood. Results are summarized in ● ▶ Table 1 .

The results of platelet function assays are summarized in ● ▶ Table 2 .

HSR tests at 0 h and 4 h showed platelet recovery of around 54 %

with no diff erences detected between time points or device.

Tests of the aggregation of PRP revealed total aggregation at

10 min was above 95 % at both time points with no diff erences

found between time or device. Upon in vitro activation with col-

lagen there was a signifi cant increase in surface expression of

CD62p (P selectin) confi rming the collected platelets are func-

tional and capable of activation at 0 and 4 h for both devices.

Discussion

▼ The current study demonstrates that both the AFC and GPS

devices produces platelet and leukocyte enriched plasma in a

closed system with functional viable platelets in volumes suita-

ble for clinical use. Although platelet concentration has been

previously reported, platelet viability has not. Additionally the

AFC performance has not been reported.

Platelet function Damage to platelets during processing can lead to the premature

release and subsequent loss of growth factors [ 1 ] . Viable plate-

lets exhibit several behaviours when activated. Some of these

behaviours can be monitored such as shape change, aggregation

and surface marker exposure. Platelet aggregation is considered

the gold standard in evaluating platelet function [ 41 ] . Mean

platelet aggregation above 95 % in this study represents the

retention of activity. Likewise HSR has been indicated as a very

sensitive marker for viability [ 22 ] , and predictor of in vivo sur-

vival [ 23 ] . During these tests platelet shape recovery was appar-

ent. Granular release is closely related to p-selectin expression

[ 18 ] . Identifying p-selectin on the surface of activated platelets

allows them to be quantitatively assessed as a percentage of all

platelets present. While absolute numbers of p-selectin assays

are shown to vary a great deal between laboratories [ 12 ] , it is a

useful measure to determine activation for matched samples

within a study centre. The results of the current study showed

that platelets could be activated beyond the resting state, indi-

cating preserved function.

The broad classifi cation of PRP encompasses a range of constitu-

ents at varying concentrations. The basic defi nition dictates that

it contains a platelet concentration that is increased above

circulating levels. Marx et al. [ 29 ] describes PRP as a platelet

concentration in a 5 mL volume with 1 000 000 platelets/μL

AFC GPS

Whole Bld c-PPP PRP PPP PRP

fi brinogen (g/L) 2.96 (0.73) 3.30 (0.73) + * 2.97 (0.65) 2.95 (0.65)* 2.94 (0.64)

pH 7.34 (0.03) 7.13 (0.03) + * 7.02 (0.06) + * 7.11 (0.04) + * 7.08 (0.05) + *

platelet concentration ( × 10^9/L) 222 (45.7) 18.7 (12.7) + * 926 (378) + * 11.0 (6.2) + * 1149 (497.7) + *

WBC ( × 10^9/L) 6.36 (1.42) 0.04 (0.135) + 32.2 (11.7) + * 0.01 (0.027) + 31.0 (8.9) + *

*diff erence compared to whole blood value, + diff erence between devices, p < 0.05

Table 1 Blood Assays. Data

summary for whole and processed

blood samples showing mean

(standard deviation).

AFC 0 h AFC 4 h GPS 0 h GPS 4 h

HSR ( %) 54.0 (5.0) 54.1 (6.6) 50.6 (6.8) 55.6 (6.1)

aggregation ( %) 95.2 (7.3) 96.5 (7.1) 97.4 (3.4) 98.5 (2.7)

p-selectin resting ( %) 7.7 (4.2) 6.4 (3.0) + 12.4 (6.5) 9.8 (2.5) +

p-selectin activated ( %) 25.4 (8.1)* 24.3 (9.2)* + 26.5 (6.1)* ̂ 31.9 (3.6)* +^

*diff erence between resting value, + diff erence between devices, ^ diff erence between time points

Table 2 Platelet Function Test-

ing. Data summary for platelet

function assays showing mean

(standard deviation).



(approximately a 4–5 fold increase). Gociman et al. [ 17 ] sum-

marized the reported platelet enrichments of 11 platelet-con-

centrating devices. These results showed fold increases ranging

from 1.34 to 6.07. In Gociman’s study the GPS produced a 4.86

fold increase on platelet count, which aligns well with the 5.19

increase seen here. The AFC was capable of increasing platelet

concentrations by a factor of 4.19 which places it roughly in the

middle of Gociman’s results. While many devices report fold

increases of 3–7 fold [ 3 , 6 , 43 ] , it is important to note that highly

concentrated platelet preparations may have an inhibitory eff ect

on healing [ 20 , 47 ] .

The devices were capable of recovering 46.4 % (AFC) and 59.8 %

(GPS) of all available platelets which is in line with previous pub-

lications. Leitner [ 26 ] compared 4 devices, and reported results

of 17 % for the Vivostat PRF Preparation Kit (Vivosolution A/S, Bir-

keroed, Denmark), 70 % for the Harvest SmartPReP 2 APC 60 Proc-

ess (Harvest Technologies Corp., Plymouth MA, USA) 67 % for the

PCCS Platelet Concentrate Collection System (3i PCCS, 3i Implant

Innovations, Palm Beach Gardens FL, USA) and 66 % for the Fibri-

net Autologous Fibrin & Platelet System (Cascade Medical Enter-

prises Ltd, Plymouth, UK). Everts et al. [ 14 ] reported results for

the Autologous Growth Factor Filter (Interpore Cross, Irvine CA,

USA) at 32 %, the Electa Cell-Separator (Sorin Group, Irandola,

Italy) at 48 % but reports a lower value for the GPS Platelet Separa-

tion System (Biomet Biologincs, Inc., Warsaw IN) at 36 %.

Although the ideal concentration of platelets within PRP remains

unknown, the eff ect of PRP is likely to be linked closely to plate-

let concentration. Leitner et al. [ 26 ] has shown that PDGF release

is closely related to platelet count. Likewise Everts et al. [ 14 ]

demonstrated the need for devices to maintain platelet viability

during preparation to maximise growth factor release upon acti-

vation when required. Regardless of the device or method used,

platelet recovery and viability are 2 important factors when

evaluating the PRP product.

Fibrin While platelet concentration and growth factor potential are

obviously essential aspects of PRP, fi brin (the activated form of

fi brinogen) is thought to directly induce angiogenesis by provid-

ing a matrix scaff old supporting cell migration and providing

chemotactic activity. The binding of growth factors such as

PDGF and VEGF to fi brin also supports wound healing [ 8 , 25 ] .

The fi brinogen concentration could therefore be an important

feature not always addressed when comparing diff erent PRP

products.

Passing the PPP repeated through the fi lter increased the con-

centration of fi brinogen, with each successive pass decreasing

the overall volume as water was removed. Our conservative

processing with the AFC showed an increased concentration

compared to whole blood. This process can be repeated any

desired number of times further increasing fi brinogen concen-

tration producing a product that could be used as a scaff old and

when added to PRP may alter the velocity of aggregation [ 24 ] ,

however, this was not examined in the current study.

pH Although there was a diff erence between the whole blood pH

and that of the processed samples, all groups were greater than

6.9 which is well above the level of 6.2 that would indicate a loss

of platelet function [ 22 , 33 ] . The increase in the acidity is likely

due to the addition of citrate to the blood from the ACD-A.

Leukocytes Platelets have a known direct and signifi cant role in immunity

and host defence against pathogenic microorganisms [ 4 , 45 ] .

Moojen, Everts et al. [ 32 ] demonstrated the antimicrobial poten-

tial of PRP in vitro. In addition to platelets, the PRP product col-

lected from the buff y coat layer has been reported to contain

around a 7-fold increase in leukocytes [ 7 ] . The output of both

the devices produced around a 5 fold increase above circulating

levels.

The collection of leukocytes with the PRP product is an area of

uncertainty. Anitua et al. [ 2 ] studied a concentrated platelet

product, Preparation Rich in Growth Factors (PRGF), which

avoids leukocyte content with the intent of avoiding the proin-

fl ammatory eff ects of leukocytes. An increase in leukocytes,

combined with the view of platelets themselves as innate

infl ammatory cells with acute host defence functions [ 16 ] ,

would suggest the PRP product may also be useful against post-

operative infections. The inclusion of leukocytes during process-

ing may also have benefi cial eff ects. Neutrophils have been

suggested to produce additional VEGF [ 48 ] , a protein known to

promote angiogenesis [ 2 , 50 ] . Autologous Conditioned Serum

(ACS) is created by incubating whole blood with glass beads, and

has been shown to increase the concentration of relevant growth

factors considerably without unwanted side eff ects in human

and animal tests [ 49 , 50 ] . A portion of the increase in growth

factor concentration may be attributable to the inclusion of leu-

kocytes during the activation process. Understanding whether

or not to include, and the extent to which WBCs are concen-

trated in the PRP is likely to be relevant for future studies. It has

been suggested that PRP products be described as Leukocyte

Platelet Rich Plasma (L-PRP), and Pure Platelet Rich Plasma

(P-PRP) to diff erentiate between the 2 PRP variations [ 11 ] .

Classifi cation It is commonly understood that not all PRPs are produced equal

[ 30 ] , and as such, PRP from diff erent methods and devices need

to be well characterised in order for the fi nal product to have the

anticipated eff ect. According to the specifi cations laid out by

Ehrenfest et al. [ 11 ] the product of the ACP device is classifi ed as

L-PRP as it contains leukocytes, just as the GPS output is. The

volume collected would be classifi ed as small ( < 25 % blood vol-

ume) except that it can be varied by adding fi brin-rich PPP in the

case of the AFC. Platelet and Leukocyte concentration were good

(40–80 % of all available) and platelets were healthy after collec-

tion. The systems we tested could be activated with any agonist

so fi brin is delivered unaltered, however the ability to concen-

trate the fi brin in the PPP with the option to add this to PRP may

alter the classifi cation.

One aspect that should be noted regarding the output of the AFC

is that the output can be purposefully modifi ed by the user by

changing the height of the collection window while drawing. The

protocol used in the present study collected a portion of the red

cell pack, the buff y coat and the plasma portion resulting from a

single spin. The PRP included some red cells which, while bio-

logically may have little impact in common use, but may be of

interest for future studies depending on the application. It is pos-

sible that the protocol could be altered to allow collections

excluding red cells, and possibly the leukocyte layer, although

this was not studied in the present study. The location of the

drawing window also dictates that a full blood draw be used with

the device so that the buff y coat layer is in the drawing area.

79Clinical Sciences


It is anticipated that the full characterisation of PRP produced by

the untested device and correlation with the existing device will

allow future studies to accurately assess what is being implanted.

Having this information widely available will also aid the practi-

tioner in determining if the output is appropriate for the needs

of their patients.

Acknowledgements

▼ The work presented in this study was supported by funding

from Circle Biologics, Minneapolis, MN. No individual author

received funding from Circle Biologics or other sources in rela-

tion to this study.

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