Leukocyte concentration and composition in platelet-rich plasma (PRP) influences the growth factor and protease concentrations

Abstract: Recently, platelet rich plasma has been popular and its use has begin on human in developed countries. Platelet rich plasma is defined as autologus blood with concentration of platelets three to five times above baseline level, which contains at least seven growth factors like Platelet Derived Growth Factor (PDGF), Platelet Derived Angiogenesis Factor (PDAF), Platelet Derived Endothelial Growth Factor (PDEGF), Transforming Growth Factor Beta (TGF- β), Insulin like Growth Factor (IGF), Fibroblast Growth Factor (FGF), and Vascular Endothelial Growth Factor (VEGF). The golden standard for reconstruction of cranial bone defects demonstrates osteoconduction scaffold, osteoinduction like growth factors, and osteogenesis. Alloplastic biomaterials have revolutionalized craniofacial reconstruction. Carbonated hydroxyapatite (CHA) has been studied for years as implant material due to its similarity with the mineral component of bone. In this study we investigated and compare the effects of PRP and CHA on bone regeneration in rat cranial defects. This was an experimental study with a true experimental design on white male rats (Rattus norvegicus). Cranial deffects of 3 mm diameter were created in rat cranium and grafted with CHA and PRP combination, CHA alone, and control. The relationships among them were analyzed by using Mann Whitney and SPSS Statistics Program Package Version 22.0. The results showed that the experimental group of 2 weeks had no different between inflammatory reaction (P = 0.119), woven bone (P = 0.094) and lamellar bone (P = 0.130). At 4 weeks,a combination of PRP and CHA showed a superior growth of lamellar bone compared to CHA (P = 0.009). Conclusion: A combination of PRP and CHA in bone regeneration showed a histological tendency toward increased bone formation. However, future investigations should be conducted in different period times.Keywords: platelet rich plasma, carbonated hydroxyapatite, cranial defectAbstrak: Plasma kaya trombosit makin banyak digunakan dalam dunia kedokteran. Di negara maju pengunaannya sudah mulai diteliti pada manusia. Plasma kaya trombosit adalah fraksi plasma darah dengan konsentrasi platelet 3-5 kali diatas nilai normal yang mengandung sekurang-kurangnya 7 faktor pertumbuhan, diantaranya Platelet Derived Growth Factor (PDGF), Platelet Derived Angiogenesis Factor (PDAF), Platelet Derived Endothelial Growth Factor (PDEGF), Transforming Growth Factor Beta (TGF- β), Insulin like Growth Factor (IGF), Fibroblast Growth Factor (FGF), dan Vascular Endothelial Growth Factor (VEGF) yang dapat meningkatkan proses osteogenesis. Karbonat hidroksiapatit adalah material pengganti tulang yang dapat mempercepat regenerasi jaringan tulang serta memiliki kandungan kalsium,fosfat dan karbonat yang mirip dengan tulang manusia. Tulang yang tumbuh pada awal berupa tulang muda yang memiliki serat kolagen yang tidak teratur dan banyak osteosit disebut tulang imatur. Tulang imatur kemudian akan diganti oleh tulang matur yang memiliki serabut kolagen yang teratur. Jenis penelitian ini ialah eksperimental pada 36 hewan coba tikus putih wistar (Rattus norvegicus). Defek kalvaria pada tikus dengan diameter 3 mm diisi sesuai perlakuan: plasma kaya trombosit dengan karbonat hidroksiapatit, karbonat apatit tunggal, dan kontrol. Plasma kaya trombosit dibuat dari autologus darah tikus yang diberi perlakuan plasma kaya trombosit serta karbonat hidroksiapatit dan karbonat apatit tunggal. Data dianalisis dengan uji Mann Whitney dan diolah dengan SPSS. Hasil penelitian memperlihatkan pada minggu ke-2, tidak terdapat perbedaan bermakna reaksi inflamasi (P = 0,119), tulang imatur (P = 0,094), dan tulang matur (P = 0,130) diantara ketiga perlakuan. Pada minggu ke-4, tulang matur yang terbentuk lebih banyak pada perlakuan plasma kaya trombosit dan karbonat hidroksiapatit (P = 0,009). Simpulan: Pemberian plasma kaya trombosit dan karbonat hidroksiapatit dapat meningkatkan proses penutupan defek tulang kepala hewan percobaan tikus.Kata kunci : plasma kaya trombosit, karbonat hidroksiapatit, defek tulang kepala.

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Platelet-Rich Plasma Powder: A New Preparation Method for the Standardization of Growth Factor Concentrations

The American Journal of Sports Medicine ◽

10.1177/0363546516674475 ◽

2016 ◽

Vol 45 (4) ◽

pp. 954-960 ◽

Cited By ~ 26

Author(s):

Matthias Kieb ◽

Frank Sander ◽

Cornelia Prinz ◽

Stefanie Adam ◽

Anett Mau-Möller ◽

...

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Clinical Application ◽

Whole Blood ◽

Sports Medicine ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Transforming Growth Factor Β1 ◽

Enzyme Linked Immunosorbent Assay ◽

Tgf Β1

Background: Platelet-rich plasma (PRP) is widely used in sports medicine. Available PRP preparations differ in white blood cell, platelet, and growth factor concentrations, making standardized research and clinical application challenging. Purpose: To characterize a newly standardized procedure for pooled PRP that provides defined growth factor concentrations. Study Design: Controlled laboratory study. Methods: A standardized growth factor preparation (lyophilized PRP powder) was prepared using 12 pooled platelet concentrates (PCs) derived from different donors via apheresis. Blood samples and commercially available PRP (SmartPrep-2) served as controls (n = 5). Baseline blood counts were analyzed. Additionally, single PCs (n = 5) were produced by standard platelet apheresis. The concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor AB (PDGF-AB), transforming growth factor β1 (TGF-β1), insulin-like growth factor 1 (IGF-1), interleukin (IL)–1α, IL-1β, and IL-1 receptor agonist (IL-1RA) were analyzed by enzyme-linked immunosorbent assay, and statistical analyses were performed using descriptive statistics, mean differences, 95% CIs, and P values (analysis of variance). Results: All growth factor preparation methods showed elevated concentrations of the growth factors VEGF, bFGF, PDGF-AB, and TGF-β1 compared with those of whole blood. Large interindividual differences were found in VEGF and bFGF concentrations. Respective values (mean ± SD in pg/mL) for whole blood, SmartPrep-2, PC, and PRP powder were as follows: VEGF (574 ± 147, 528 ± 233, 1087 ± 535, and 1722), bFGF (198 ± 164, 410 ± 259, 151 ± 99, and 542), PDGF-AB (2394 ± 451, 17,846 ± 3087, 18,461 ± 4455, and 23,023), and TGF-β1 (14,356 ± 4527, 77,533 ± 13,918, 68,582 ± 7388, and 87,495). IGF-1 was found in SmartPrep-2 (1539 ± 348 pg/mL). For PC (2266 ± 485 pg/mL), IGF-1 was measured at the same levels of whole blood (2317 ± 711 pg/mL) but was not detectable in PRP powder. IL-1α was detectable in whole blood (111 ± 35 pg/mL) and SmartPrep-2 (119 ± 44 pg/mL). Conclusion: Problems with PRP such as absent standardization, lack of consistency among studies, and black box dosage could be solved by using characterized PRP powder made by pooling and lyophilizing multiple PCs. The new PRP powder opens up new possibilities for PRP research as well as for the treatment of patients. Clinical Relevance: The preparation of pooled PRP by means of lyophilization may allow physicians to apply a defined amount of growth factors by using a defined amount of PRP powder. Moreover, PRP powder as a dry substance with no need for centrifugation could become ubiquitously available, thus saving time and staff resources in clinical practice. However, before transferring the results of this basic science study to clinical application, regulatory issues have to be cleared.

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