Beruflich Dokumente
Kultur Dokumente
Schattauer 2011
Original Research
of Small Animal Medicine, University of Leipzig, Leipzig, Germany; 2Institute of Physiological Chemistry,
University of Leipzig, Leipzig, Germany; 3Large Animal Clinic for Surgery, University of Leipzig, Leipzig, Germany
Keywords
Platelet-rich plasma, dog, thrombocyte,
growth factor
Summary
Objectives: To report the concentration of
blood cells and selected growth factors in canine autologous conditioned plasma (ACP).
Methods: The density of blood cells in whole
blood (WB), ACP and standard plasma preparation (SP) of 10 healthy mature dogs was
determined. In both ACP and SP, the concentration of insulin-like growth factor-1 (IGF-1),
epidermal growth factor, vascular endothelial
growth factor, platelet-derived growth factorAA, platelet-derived growth factor-AB, platelet-derived growth factor-BB, transforming
growth factor-1 (TGF-1), and transforming
growth factor-2 was measured using the
ELISA technique. In another ten dogs, ACP was
prepared using an ultra-soft spinning protocol, and again blood cell density was compared to that obtained in WB.
Results: The density of platelets in ACP was
significantly higher than that in SP (p =
Correspondence to:
Dr. Peter Bttcher, Dr. med. vet., Dipl. ECVS
Klinik fr Kleintiere
Universitt Leipzig
An den Tierkliniken 23
D-04103 Leipzig
Germany
E-mail: boettcher@kleintierklinik.uni-leipzig.de
Phone: +49 341 97 38 700
Fax: +49 341 97 38 799
Introduction
For treatment of various acute and chronic
sport injuries, platelet-rich concentrates
(PRC) have incrementally gained in importance in both humans and animals (14).
0.0002), but there was not any significant difference between ACP and WB, nor between
WB and ACP prepared using softer centrifugations. Interestingly, only for IGF-1, PDGFBB, and TGF-1 could reliable measurements
be obtained, showing a significant increase in
PDGF-BB and TGF-1 concentrations in ACP
compared to SP (p = 0.001, p = 0.0028). Regarding IGF-1 content, there was not any significant difference between ACP and SP.
Clinical significance: Canine ACP prepared
according to the manufacturers recommendations, or by using a softer spin does not
show the same specifications as human ACP,
which shows a doubling in platelet count
compared to WB. Even though canine ACP has
a similar number of platelets per injected volume and consequently, probably the same
amount of injected growth factors than WB,
application of canine ACP would not be associated with the proinflammatory potential
reported for WB, as it is almost free of erythrocytes and nucleated cells.
Financial support:
Arthrex provided financial support for the ELISA.
Vet Comp Orthop Traumatol 2011; 24: 122125
doi:10.3415/VCOT-10-04-0064
Received: April 20, 2010
Accepted: September 6, 2010
Pre-published online: January 11, 2011
Data analysis
Descriptive statistics were performed using
commercial softwaref testing for normaldistribution of data using the D'Agostinoc
d
e
Results
Autologous conditioned plasma of group 1
prepared at 350 g showed only marginal
concentrations of leukocytes (median 0.10
G/l [G = Giga]; IQR: 0.03 0.25) and erythrocytes (median 0.01 T/l [T = Tera]; IQR:
0.00 0.01) (!Table 1). Manually counted
concentrations of platelets highly correlated with the results obtained using the
cell counter (Spearmans rho = 0.948; p
<0.001 ; R2 = 0.9754), thus only platelet
concentrations obtained using the cell
counter were finally analysed and reported
hereby.
Median platelet count in group 1 for
WB, ACP, and SP are reported in !Table 1.
Calculation of the Wilcoxon rank sum test
did not show any significant increase in
platelet count in ACP (293.50 G/l) compared to WB (271.50 G/l) (p = 0.3594).
With a more than 10-fold increase, concentration of platelets in ACP was significantly
higher than in SP (p = 0.002) in group 1.
In group 2, lowering the spin speed at
which ACP was centrifuged while increasing the duration of centrifugation (700
rpm [75 g] for 15 minutes) concurrently
did not result in any significant change in
the median platelet concentration of ACP
(277.00 G/l; IQR: 197.75 395.00) or
WB (255.50 G/l; IQR: 205.50 273.50)
Schattauer 2011
123
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Cell density within whole blood, autologous conditioned plasma (ACP), and standard plasma for group 1 (n = 10) and within whole blood and
ACP for group 2 (n = 10). All values are expressed as median and associated interquartile range (IQR).
Whole blood
PLT (G/l)
Standard plasma#
WBC (G/l)
PLT
(G/l)
WBC
(G/l)
RBC
(T/l)
PLT
(G/l)
WBC
(G/l)
RBC
(T/l)
Group 1
271.50a
9.35
(244.75-338.75) (8.45-9.95)
6.78
(6.65-7.17)
293.50a
0.10
(221.00-321.75) (0.03-0.25)
0.01
29.00b
0.00
(0.00-0.01) (23.00-55.25) (0.00-0.00)
0.00
(0.00-0.00)
Group 2
10.25
255.50a
(205.50-273.50) (8.70-11.50)
6.08
(5.91-6.54)
277.00a
0.00
(197.75-395.00) (0.00-0.35)
0.02
(0.01-0.05)
Key: PLT = platelets; WBC = white blood cells; RBC = red blood cells; G = Giga; T = Tera; *= Autologous conditioned plasma was centrifuged at 350 g
for five minutes for group 1, and at 75 g for 15 minutes for group 2; # = Standard plasma was centrifuged at 2000 g for 10 minutes; a, b = Concentrations
of platelets having different superscripts are significantly different (p 0.05).
Discussion
This is the first study as far as we know which
evaluates platelet count and growth factor
concentration in canine ACP. Even though
the presented findings have to be regarded as
preliminary due to the limited sample size
and unexpected shortcomings during
Table 2 Concentration of growth factors measured in standard citrate plasma and autologous
conditioned plasma using Quantikine ELISA kitsd. Values are expressed as mean standard deviation.
Autologous
conditioned
plasma*
Standard
plasma#
Factor of
enrichment
68.5 35.6
67.9 32.1
1.0
No signal
No signal
No signal
No signal
No signal
No signal
No signal
No signal
251.0 132.8
1239.9 590.8
>2000.0
(not available)
>1.6
No signal
No signal
measurement of growth factor concentrations, the study may serve as a guide for
clinicians, allowing them to appreciate the
potential biological character of ACP. When
prepared as currently recommended by the
manufacturer using 350 g for five minutes,
ACP did not reach significantly higher platelet counts than unprocessed whole blood.
This contrasts with the results obtained
when using human blood samples, in which
a two-fold increase in platelet count could be
detected (11). When comparing conventionally prepared canine plasma to canine
ACP, platelet count in ACP was about
10-fold higher. This significant enrichment
in platelets is the reason for the elevated concentration of -granule-derived growth factors observed in canine ACP compared to SP,
supporting the previously reported correlation between platelet count and concentration of platelet-derived growth factors
(8). Insulin-like growth factor-1 was not
elevated in ACP compared to SP, which is an
expected finding when preparing PRC (11,
20). Insulin-like growth factor-1 is primarily
excreted by the liver into the blood plasma,
being independent from the present number
of platelets (21). The inclusion of IGF-1 into
to the study protocol was thought to serve as
a negative control, differentiating platelet related and unrelated growth factor enrichment.
The problems encountered during
growth factor assay, which resulted in unreliable measurements for EGF, VEGFcanine, PDGF-AA, PDGF-AB and TGF-2
both in SP and ACP, may have been related
to species differences when using human
ELISA kits as well as when using kits not
Schattauer 2011
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