5 1016NelsonTatsui Effect Prp on Apoptosis

8/14/2019 5 1016NelsonTatsui Effect Prp on Apoptosis

1/12

EFFECT OF PLATELET-RICH PLASMA ON IMPACT-INDUCED

CHONDROCYTE APOPTOSIS: EXPERIMENTAL STUDY

Mrcia Uchoa de Rezende

Ronald Bispo Barreto Silva

Ana Cristina Ferreira Bassit

Nelson Hidekazu Tatsui

David Sadigursky

Moyses Sadigurky

Medical Investigation Laboratory-41

Institute of Orthopedics and Traumatology

Medical School of Sao Paulo University


2/12

Introduo

[email protected]

Articular cartilage has poor

reparative potential, but the

reasons for this are not fully

understood

Osteoarthritis Cartilage 2001;9:712-9

J Orthop Sports Phys Ther 1998;28:192-202

Chondrocyte apoptosis has been

shown to occur in osteoarthritis

and in response to mechanical

injury in vitro studiesJ Bone Joint Surg 2001:83-A Suppl 2: 19-21

Arthritis Rheum 1998;41:284-9

J Orthop Res 2001;19:703-11


3/12

Introduo

[email protected]

Several studies establish direct or indirect evidence that

platelet-derived products have a substantial role in tissue

regenerationTransfusion 2005;45:1759-67

Curr Opin Hematol 2005;12:473-9

Recent investigations have demonstrated that blockade of

apoptosis by pharmacologic agents decreases cell death and

improves survival

Arthritis Rheum 2006;54:1814-21

Ann Rheum Dis 2005;64:89-94

Int Orthop 2007;31:773-81


4/12

Objetivo

[email protected]

The objective of this study is to evaluate if intra-

articular (IA) injection of platelet rich plasma(PRP) reduces impact-induced chondrocyte apoptosis

following a blunt trauma


5/12

Materiais e Mtodos

ethical principles of the:

COBEA (Brazilian College of Animal Experimentation) American Veterinary Medical

Association (AVMA)

Institute of Animal Care and Use Committee (IACUC)

02 Mature New Zealand white rabbits were used in the present

study

Mazires contusion model of osteoarthritis was reproduced 3

times in each kneeMazieres B, Berdah L, Thiechart M, Viguier G. Diacetylrhein on a postcontusion model of

experimental osteoarthritis in the rabbit. Rev Rhum Ed Fr 1993;60:77S-81S

each rabbit received 1ml injection of human-derived PRP

(Apheresis Technology Haemonetics Corporation) in the right

knees (now-called knees 1 and 3) and 1ml injection of normal

saline solution (NaCl 0.9%) in the left knees (now-called

knees 2 and 4)


6/12

Materiais e Mtodos

[email protected]

Ten days after knee contusion the animals were euthanized.

This time point was selected to focus on the early phase of

the posttraumatic apoptosisJ Bone Joint Surg Am 2001;83-A Suppl 2:19-21

Fresh cartilage fragments were harvested from patella, and

both femoral condyles

Ultrathin sections (60 nm) were cut and stained with uranyl

acetate and lead citrate and examined on a TEM (transmission

electron microscope (JEOL JEM-1010,Olympus,Japan)


7/12

Materiais e Mtodos

[email protected]

Statistical Analysis

The statistical significance of the differences inapoptosis was analyzed by qui-square test. The

estimation of the sample size resulted in 142 cells for

each group.

The technician responsible to prepare the cartilagespecimens and the pathologist who was responsible to

count cells were blind to the groups they were studying

(double-blind methodology). For the histological study,

each cartilage specimen was divided in random TEM fields.

In these fields,normals and apoptotics chondrocytes were

counted


8/12

Resultados

[email protected]

Seven to ten fields (average 8,75)of TEM wereobtained for each knee

Figure 1. Transmission Electron Microscopy of knee cartilage specimens. White

arrow, normal chondrocyte with abundant cytoplasm and normal cellular and nuclear

morphology. Black arrow, chondrocyte demonstrating changes consistent with

apoptosis including nuclear fragmentation, cell shrinkage and cytoplasmic membrane

blebbing.


9/12

Resultados

[email protected]

70,4012537884

48,3612263593

56,673013172

47,6210555501

% apoptosisTotal of CellsNormalApoptoticKnee

Table 1. Number of chondrocytes observed in each knee of the study

Knees treated with PRP (Knees 1 and 3), showed 47.62%

(50/105) and 48.36% (59/122) of apoptosis, respectively. In

contrast, in the control group (Knees 2 and 4), the rates of

apoptosis were 56.67% (17/30) and 70.40% (88/125),

respectively (Table 1)


10/12

Resultados

[email protected]

PRP intra-articular injection was associated with a

significantly decreased number of chondrocyte death 48.02% in

respect to 67.74% in the control group (p


11/12

Discusso

[email protected]

In vitro studies showed that PDGF are number of platelets/ml

dependent. ONeill et al. showed that the method by which

the PRP is collected affects the volume of PRP and the

concentration of platelets achieved. Because of this number

related efficacy we used 1ml of PRP obtained by apheresistechnology (MCS plus 9000)Bone 2004;34:665-71

Vox Sang 2001,81172-5

DLima et al. have shown the progressive increase in the

levels of apoptotic cells after injuries, offering a

potentially therapeutic windowOsteoarthritis Cartilage 2001;9:712-9

J Bone Joint Surg Am 2001;83-A Suppl 2:19-21

J Bone Joint Surg Am 2001;83-A Suppl 2:25-6


12/12

Discusso

[email protected]

If one single injection of PRP can significantly reduce

the number of apoptotic cells, increased dose and

frequency should be studied to verify a more expressive

chondroprotective action of PDGF post-trauma

Concluso

Immediately post-traumatic intra-articular injection ofPRP reduces impact-induced chondrocyte apoptosis in

rabbits.

Documents

5 1016NelsonTatsui Effect Prp on Apoptosis