The inhibitory effect of platelet rich plasma mesotheraphy on botulinum toxin type-A induced skeletal muscle paralysis

The Inhibitory Effect of Platelet Rich Plasma Mesotheraphy on Botulinum Toxin Type-A induced Skeletal Muscle Paralysis Platelet Rich Plasma (PRP) is a platelet concentrate that is produced from autologous blood by centrifugation. PRP is used in various areas of medicine because of its wound healing and cell proliferation effects. These effects depend on growth factors in ? granules of platelets. They also have various coagulation factors, proteins and enzymes. Botulinum toxin type-A (BTxA) is a neurotoxin which have a metalloproteinase activity. BTxA causes muscle paralysis as it breaks SNAP-25 perotein in nerve terminal and prevents acetyl-choline release to neuromuscular juction. Aim of this study is to investigate the effect of PRP on BTxA induced skeletal muscle paralysis. Anterior auricular muscle (AAM) model was used in twenty-four New Zeland Rabbit which had been studied in 4 groups. Right and left AAM and skin over the AAM?s were used for injection procedures. Autologous PRP was prepared with ACR Regenkit® for all rabbits. BOTOX was used for botulinum toxin type-A injections. 2,5U BTxA was injected to bilateral AAM intramuscularly and 1cc autologous PRP intradermally (PRP mesotheraphy) in Group I (n=6). Group II (n=6) was designed as control group of group I and 2,5U BTxA was injected to bilateral AAM intramuscularly and 1cc %0.9 NaCl that was centrifuged in ACR Regenkit was injected intradermally. Group III (n=6) was designed as in-vitro mixture group. 2,5 U BTxA (0,05ml) and 0,05ml PRP was mixed in laboratory tube and then was given intramuscularly to right AAM of rabbits. 2,5 UBTxA (0,05ml) and equal volume %0.9 NaCl was mixed and injected to left AAM. Same procedure was done in Group IV (n=6) except 2,5 U BTxA and 0,05ml %0.9 NaCl that was centrifuged in ACR Regenkit was mixed and used on right side. Electroneuromyolographic (ENMG) study was done before all procedures. Right and left AAM action potantial amplitude was noted. Fourteen days after the procedures visual study according to position of rabbits was done. Then ENGM study was repeated. Also genetical evaluation was done in muscle samples on 14th day. for SNAP-25 protein m-RNA was evaluated with Quantitative Real-Time PCR. In visual and ENGM studies it is observed that there was less BTxA activity on right side muscles of Group I (PRP mesotherapy group). These differences were statistically significant. No significant differences were found in Group II. When the findings of Group III (PRP in-vitro mixture group) were evaluated it was seen that the reduction rate of action potential amplitude of right AAM was significantly lower than left side. There was no significant differences in Group IV. SNAP-25 mRNA expression was observed in all muscle samples. No significant difference was found between groups and right/left side AAM?s of rabbits. It is tought that maybe there was a difference in posttranslational modification level. But we could not use the technique that unbroken SNAP-25 proteins could be detected. In the light of these findings we found that PRP have an inhibitory effect on BTxA activity. We observed this effect after PRP mesotheraphy and also in-vitro PRP/BTxA mixture. We could not explain the exact mechanism underlying this interaction but we assumed that the ?-2 macroglobuline in platelet?s ?-granüle and serum may have some inhibitory effect on BTxA because ?-2 macroglobuline had a general protease inhibitör activity. New studies are designed to investigate this interaction between PRP and BTxA.