Production of Platelet-Rich Gel Using Allogeneic Platelets Isolated from Donor Whole Blood
The aim of the study was to perform a morphofunctional analysis of allogeneic platelets isolated from donor whole blood and to assess the possibility of producing platelet-rich gel based on them.
Materials and Methods. The study investigated donor blood platelets, platelets collected from donors via apheresis, and platelets isolated from donor whole blood (platelet-leukocyte concentrates, PLC). Platelet quality was assessed before and after centrifugation at 2500–4000 g, as well as the feasibility of producing platelet gel from platelet-rich plasma isolated from whole blood. Morphofunctional analysis of platelets was performed using an original method based on the examination of vitally stained cells by fluorescence microscopy. The cytokine profile in the platelet concentrates isolated from whole blood was evaluated using multiplex analysis.
Results. Following centrifugation at 2500–4000 g, the platelet population exhibited an increased number of platelets with damaged membranes, procoagulant platelets, and platelets prone to spontaneous activation. Centrifugation at 2500–2700 g was less damaging than centrifugation at 2701–4000 g and allowed to preserve a significant volume of growth factors within the platelets. Platelet-rich plasma isolated from PLC can be used to produce platelet gel and thrombofibrin clot in vitro at 20–22°C without the use of platelet activation inducers. The most effective PLC samples for producing platelet gel were those without platelet conglomerates, containing over 20% platelets with granules and less than 20% platelets with damaged membranes, with a level of procoagulant platelets below 5%, and with an increased or high adhesion rate of platelets with granules to glass. The use of platelets from allogeneic PLC enables the production of platelet gel within 20–30 min.
Conclusion. There was developed a method for producing platelet gel and a thrombofibrin clot at 20–22°C based on allogeneic platelets isolated from donor whole blood. These materials can be used as wound dressings, applicative biological constructs for treating tissue defects of various origins, and in the creation of composite biological designs intended for use in regenerative medicine.
- Golebiewska E.M., Poole AW. Secrets of platelet exocytosis — what do we really know about platelet secretion mechanisms? Br J Haematol 2013; 165(2): 204–216, https://doi.org/10.1111/bjh.12682.
- Amable P.R., Carias R.B., Teixeira M.V., da Cruz Pacheco I., Corrêa do Amaral R.J., Granjeiro J.M., Borojevic R. Platelet-rich plasma preparation for regenerative medicine: optimization and quantification of cytokines and growth factors. Stem Cell Res Ther 2013; 4(3): 67, https://doi.org/10.1186/scrt218.
- Palumbo V.D., Rizzuto S., Damiano G., Fazzotta S., Gottardo A., Mazzola G., Lo Monte A.I. Use of platelet concentrate gel in second-intention wound healing: a case report. J Med Case Rep 2021; 15(1): 85, https://doi.org/10.1186/s13256-020-02649-6.
- Everts P.A., Lana J.F., Alexander R.W., Dallo I., Kon E., Ambach M.A., van Zundert A., Podesta L. Profound properties of protein-rich, platelet-rich plasma matrices as novel, multi-purpose biological platforms in tissue repair, regeneration, and wound healing. Int J Mol Sci 2024; 25(14): 7914, https://doi.org/10.3390/ijms25147914.
- Makarov M.S. Physiological and prognostic value of platelets without granules. Medicinskij alfavit 2018; 3(26): 32–36.
- Rozhkov E.V., Kozhemyako O.V., Ponasenko M.A., Karaseva I.A., Rozhkova N.S., Madzaev S.R., Zhiburt E.B. Improvement of pooled pathogen-reduced platelets concentrate production. Transfuziologiya 2022; 23(1): 16–21.
- Akbarzadeh S., McKenzie M.B., Rahman M.M., Cleland H. Аllogeneic platelet-rich plasma: is it safe and effective for wound repair? Eur Surg Res 2021; 62(1): 1–9, https://doi.org/10.1159/000514223.
- Asadi M., Alamdari D.H., Rahimi H.R., Aliakbarian M., Jangjoo A., Abdollahi A., Bahar M.M., Azadmand A., Forghani N., Sadegh M.N., Khayamy M.E., Seifalian A. Treatment of life-threatening wounds with a combination of allogenic platelet-rich plasma, fibrin glue and collagen matrix, and a literature review. Exp Ther Med 2014; 8(2): 423–429, https://doi.org/10.3892/etm.2014.1747.
- Wang S., Ding W., Du Y., Qi Q., Luo K., Luan J., Shen Y., Chen B. Allogeneic platelet gel therapy for refractory abdominal wound healing: a preliminary study. Adv Clin Exp Med 2023; 32(8): 865–872, https://doi.org/10.17219/acem/159088.
- Fujioka-Kobayashi M., Schaller B., Mourão C.F.A.B., Zhang Y., Sculean A., Miron R.J. Biological characterization of an injectable platelet-rich fibrin mixture consisting of autologous albumin gel and liquid platelet-rich fibrin (Alb-PRF). Platelets 2021; 32(1): 74–81, https://doi.org/10.1080/09537104.2020.1717455.
- Fan Y., Perez K., Dym H. Сlinical uses of platelet-rich fibrin in oral and maxillofacial surgery. Dent Clin North Am 2020; 64(2): 291–303, https://doi.org/10.1016/j.cden.2019.12.012.
- Makarov M.S., Khubutia M.Sh., Khvatov V.B., Vysochin I.V., Kobzeva E.N., Konyushko O.I. A method for assessing the morphofunctional status of human platelets. Patent RU 2485502. 2013.
- Podoplelova N.A., Sveshnikova A.N., Kotova Y.N., Eckly A., Receveur N., Nechipurenko D.Y., Obydennyi S.I., Kireev I.I., Gachet C., Ataullakhanov F.I., Mangin P.H., Panteleev M.A. Coagulation factors bound to procoagulant platelets concentrate in cap structures to promote clotting. Blood 2016; 128(13): 1745–1755, https://doi.org/10.1182/blood-2016-02-696898.
- Makarov M.S., Borovkova N.V., Khvatov V.B., Kobzeva E.N. The effect of centrifugation on the biological usefulness of human platelets. Vestnik sluzby krovi Rossii 2015; 1: 41–44.
- Denorme F., Campbell R.A. Procoagulant platelets: novel players in thromboinflammation. Am J Physiol Cell Physiol 2022; 323(4): C951–C958, https://doi.org/10.1152/ajpcell.00252.2022.
- Italiano J.E. Jr, Mairuhu A.T., Flaumenhaft R. Clinical relevance of microparticles from platelets and megakaryocytes. Curr Opin Hematol 2010; 17(6): 578–584, https://doi.org/10.1097/MOH.0b013e32833e77ee.
- Makarov M.S., Storozheva M.V., Borovkova N.V., Ponomarev I.N. A method for preparing a thrombofibrin clot with growth-stimulating properties. Patent RU 2679616. 2019.
- Santhakumar M., Yayathi S., Retnakumari N. A clinicoradiographic comparison of the effects of platelet-rich fibrin gel and platelet-rich fibrin membrane as scaffolds in the apexification treatment of young permanent teeth. J Indian Soc Pedod Prev Dent 2018; 36(1): 65–70, https://doi.org/10.4103/JISPPD.JISPPD_180_17.
- Nadra M., Niu W., Kurisawa M., Rousson D., Spector M. Platelet-rich plasma lysate-incorporating gelatin hydrogel as a scaffold for bone reconstruction. Bioengineering (Basel) 2022; 9(10): 513, https://doi.org/10.3390/bioengineering9100513.
- Tang S., Wang L., Zhang Y., Zhang F. A biomimetic platelet-rich plasma-based interpenetrating network printable hydrogel for bone regeneration. Front Bioeng Biotechnol 2022; 10: 887454, https://doi.org/10.3389/fbioe.2022.887454.
- Sitkova E.S., Dragunova M.A., Ogurkova O.N., Smorgon A.V., Moskovskikh T.V., Batalov R.E., Suslova T.E. Spontaneous and stimulated platelet aggregation activity in patients with atrial fibrillation and thrombotic complications. Siberian Journal of Clinical and Experimental Medicine 2023; 38(4): 116–124, https://doi.org/10.29001/2073-8552-2023-38-4-116-124.
- Moskalensky A.E., Litvinenko A.L. The platelet shape change: biophysical basis and physiological consequences. Platelets 2019; 30(5): 543–548, https://doi.org/10.1080/09537104.2018.1514109.
- Murphy D.D., Reddy E.C., Moran N., O'Neill S. Regulation of platelet activity in a changing redox environment. Antioxid Redox Signal 2014; 20(13): 2074–2089, https://doi.org/10.1089/ars.2013.5698.









