Tag Archives: platelet rich plasma

Administration of platelet-rich plasma or concentrated bone marrow aspirate after mechanically induced ischemia improves biochemical parameters in skeletal muscle

А. Т. Pidlisetsky1, G. V. Kosiakova2, T. M. Goridko2, A. G. Berdyschev2,
O. F. Meged2, S. I. Savosko3*, О. V. Dolgopolov4

1Lviv Regional Hospital for War Veterans and Repressed named after Yu. Lypa, Ukraine;
2Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
3Bogomolets National Medical University, Kyiv, Ukraine;
4SI “Institute of Traumatology and Orthopedics of NAMS of Ukraine”, Kyiv;
*e-mail: savosko_s@ukr.net

Received: 16 September 2020; Accepted: 17 May 2021

Ischemia is known to cause structural and functional damage to skeletal muscle. In this research platelet-rich plasma (PRP) and cell  bone marrow aspirate (CBMA) which are considered to be promising in  regenerative medicine were used to improve the animal muscles condition after ischemia. Mechanically induced ischemia (MII) of the rabbit skeletal muscles was caused by applying a tourniquet to the hind limb for 6 hours.  At 6 h after tourniquet was removed the rabbit PRP or CBMA solutions were injected into the tibia muscle and animals were divided into experimental groups (n = 15): 1 – ischemia + saline, 2 – ischemia + PRP, 3 – ischemia + CBMA. Intact animals were the control group (n = 5). Animals were removed from the experiment on 5, 15, and 30th days of reperfusion  after ischemia. A progressive hypotrophy, muscle fibers destruction and leukocyte infiltration was observed after tibia muscle MII. The ischemic state was accompanied by catalase and superoxide dismutase activation, persistent NO2 overproduction and a significant decrease in glutathione peroxidase activity in the muscle homogenates. It was shown that after PRP or CBMA local injection into the ischemic muscle catalase activity remained at the elevated level, SOD activity approached the control level and the most pronounced effects were NO2 level decrease and glutathioperoxidase activity restoration. After CBMA injection angiogenesis activation and the increase in the number of nuclei in the muscle fibrils were observed. The obtained data allow to expand the range of biochemical parameters for assessing the muscles condition after ischemia and indicate a positive effect of platelet-rich plasma and cell bone marrow aspirate injection on skeletal muscle regeneration.

Preparation of highly-concentrated autologous platelet-rich plasma for biomedical use

V. Chernyshenko1, K. Shteinberg2, N. Lugovska1, M. Ryzhykova1,
T. Platonova1, D. Korolova1, E. Lugovskoy1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: bio.cherv@gmail.com;
2‘Dr. Zapolska Clinic’, Kyiv, Ukraine

Received: 21 December 2018; Accepted: 20 March 2019

Cell therapy with platelets is a widely accepted approach for wound healing and tissue regeneration in medicine. However, with most available methods poorly concentrated platelet suspensions (up to 0.3∙106 1/µl) or suspensions of mostly inactivated or lost platelets are obtained. In this study, we aimed to develop a simple and effective method for preparing a suspension of native and resting platelets with over 1∙106 1/µl. Platelet-rich plasma (PRP) was obtained from fresh blood of healthy donors (n = 5) collected using different amounts of heparin as the anticoagulant. Samples of PRP were spun down and re-suspended in auto­logous blood plasma. Count and vitality of platelets in each sample were determined by aggregation study on the Solar AP2110 aggregometer. Platelet shape and cytoplasmic granularity that indicate the nativity of platelets were monitored on the COULTER EPICS XL Flow Cytometer. This study of aggregation of platelets in PRP obtained using various amounts of heparin allowed us to reduce final concentrations to the amount that effectively prevented clotting and did not affect platelet reactivi­ty (5 U/ml). PRP concentrated 5 times with a total concentration of cells of 1∙106 1/µl was able to be activated by adenosine diphosphate (ADP) (aggregation rate 54 ± 7%). The amount of cells with altered shape and granularity in concentrated suspension was not higher than 20%. This finding means that the platelets would still be able to release a number of growth factors and other biologically active compounds after stimulation or injection into tissue during cell therapy. The decrease in heparin concentrations also minimizes haemorrhage in the injection site supporting biomedical use of the suspension. A simple and effective method for preparation of highly-concentrated PRP (1.2∙106 1/µl) for biomedical use was developed. Aggregometry and flow cytometry proved that obtained platelets were resting and able to be activated. Being autologous, the preparation can be widely used for cell therapy without additional precautions.