STUDY PROTOCOL FOR JOINT PRP KIT SYSTEM VALIDATION

Transfusion Medicine Center ASO SS Antonio e Biagio - Alessandria (Italy)
Regenerative Medicine and Blood components production Laboratory

" The system has demonstrated a good platelet recovery performance (average concentration equal to 5.4 times the baseline), 99% of the tests satisfy the requirements of current DM about platelets concentration (Italian DM 02 November 2015: 800 -1,200 for 103 / mm3). Platelets are recovered quickly and easily from buffy coat isolation after centrifugation, this also include the collection of leukocytes (on average 13.7 x 103 / mm3, an average concentration equal to 2.4 times the basaline) with greater presence of mononuclear cells (61% lymphocytes-monocytes) compared to baseline, turning this product into a Leuco-PRP (L-PRP), according with current literature classifications. "

Blood cells count

VIEW THE TABLE
BASELINE L-PRP PLASMA
PLT
103/mm3 		WBC
103/mm3 		PLT*
103/mm3 		PLT
recovery conc fold 		WBC
103/mm3 		WBCrecovery
conc fold 		PLT
103/mm3 		WBC
103/mm3
group A 160
168
166 		5,6
5,4
3,9 		1034
1079
700 		6,4
6,4
4,2 		8,3
11,3
14 		1,5
2,1
3,5 		70
66
137 		0,4
0,3
0,4
group B 186
186
197 		5,0
4,7
4,6 		1615
1040
914 		8,6
5,6
4,6 		13
12,6
12,1 		2,3
2,7
2,6 		118
148
122 		0,1
0,1
0,1
group C 296
257
251 		3,7
5,4
5,0 		1486
1146
1123 		5
4,5
4,5 		17
19
16 		4,5
3,5
3,2 		97
170
118 		0,1
0,2
0

* Benchmark value of Italian DM 02.11.2015: 800 -1.200 per 103/mm3

" The IL-1 ra protein is the natural receptor antagonist of interleukin 1 (classified as a primary inflammatory cytokine) and is mainly produced by polymorphonuclear cells. The presence of leukocytes into this final PRP product, in particular the monocyte-macrophage and lymphocyte component (mononuclear cells - PMN), increase the IL-1 ra concentration. Normally, in the humoral environment, it is present in low quantities (normal value in plasma: 105-1193 pg/mL), however its quantity increases as the concentration of mononuclear cells (PMN) increases. Therefore IL-1 ra (pg/mL) was studied as probable anti-inflammatory activity index of PRP produced with JOINT PRP KIT system.

IL-1 ra dosage

VIEW THE TABLE
WBC
x 103/mm3		 PMN % PRP
IL-1 ra pg/mL		 Plasma
IL-1 ra pg/mL		 V.N.
group A 8,3
11,3
14 		50% 3691
3738
4075 		401
423
596 		105 - 1193
pg/mL
group B 13
12,6
12,1 		61% 3514
3625
3611 		391
457
426
group C 17
19
16 		61% 3681
3758
3622 		402
487
418

* Benchmark value

In addition to the right platelet concentration, leukocytes provide several inflammation antagonist molecules in the final product, in particular the mononuclear component releases an abundant quantity of IL-1 ra (at least 6 times greater than the plasma value). This exogenous quantity of autologous IL-1 ra administered in local site could strongly prevent the excess of IL-1 produced in a damaged tissue. The presence in final JOINT PRP KIT product of molecules with both anti-inflammatory (leukocytes) and repairing/regenerative (platelets) activity have a 360 ° biological action to induce lesions healing at local level."