Abstract
The aim: to develop, substantiate an effective and safe technology for producing PRP (platelet rich plasma). To quantify the substrate based on the recommended centrifugation protocols. Materials and methods: the effectiveness of the original harvesting protocol was evaluated by quantifying the number of platelets. The proposed technique is formed basing on the basic principles of double centrifugation of whole blood in test tubes with anticoagulant, separation with the release of a plasma layer with a high content of platelets. The centrifuging mode for quantifying the effectiveness of the substrate was selected according to recommendations based on a study confirming maximum efficiency (160g×10min + 250g×15min). For quantitative evaluation, blood was collected from 10 healthy volunteers (7 men, 3 women) with an average age of 26.0±2.6, and centrifuged in standard mode. Quantitative evaluation of platelets of whole blood and the obtained PRP substrate was carried out with a semi-automatic analyzer. Results: the proposed technique is based on the use as a container for centrifuging a syringe with a LuerLock design, which is hermetically sealed with a congruent plug, adapted by the external size of the centrifuge rotor bowl. Phase selection after centrifugation was performed by aspiration of the syringe contents after centrifugation is performed through a three-way valve. The substrate was obtained by repeated centrifugation of the contents, which allows obtaining a variable volume and platelet concentration in PRP. The amount of platelets (PLT) of whole blood is 227.0±57.0 thousand per ml. PLT PRP 945.0±279.0 thousand per ml. Conclusions: the proposed method of separation of whole blood with the release of the platelet rich plasma demonstrates high efficiency, which corresponds to the level of increasing the number of platelets in reducing the volume at the level of the best ready-made solutions. The equipment is economical and does not require highly specialized equipment and consumables. The proposed technique provides a wide choice to the performer in the received volume of the substrate.
Highlights
At the present stage of development of medicine, platelet rich plasma (PRP) – based plasma products, that have found application and have become firmly established in clinical practice in many medical fields [1, 2]
In a patient by venipuncture of the peripheral or central vein, blood is collected in a syringe with a LuerLock attachment mechanism filled in 10 % of the volume with a solution of the anticoagulant CITRAT glucose solution A (ACD-A) and filled to the full volume with the patient’s blood after which the syringe is sealed under aseptic conditions tightly and congruently to LuerLock stub CombiStopper
Formulated protocol: In a patient by venipuncture of the peripheral or central vein, blood is collected in a syringe with a LuerLock attachment mechanism filled in 10 % of the volume with a solution of the anticoagulant CITRAT glucose solution A (ACD-A) and filled to the full volume with the patient’s blood after which the syringe is sealed under aseptic conditions tightly and congruently to LuerLock stub CombiStoper
Summary
At the present stage of development of medicine, platelet rich plasma (PRP) – based plasma products, that have found application and have become firmly established in clinical practice in many medical fields [1, 2]. The effectiveness according to different authors varies significantly and depends on the quantitative characteristics, the main and affordable rapid assessment of which is the number of platelets per unit volume [10, 11]. This indicator determines the compliance of PRP with standardized criteria as well as the expected clinical effect. Other systems for obtaining PRP differing in the design of the container with a more complex device, allowing the selection of separated phases to be performed according to the standard procedure are less demanding on the skills of the contractor, but they are expensive [15, 16]. It is difficult to isolate the optimal variant of the PRP production system taking into account the need for volume, as well as the substantial cost of centrifuges and consumable sets for most systems [15, 17]
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