Platelet-rich plasma: quantification of growth factor levels and the effect on growth and differentiation of rat bone marrow cells.
Platelet-rich plasma (PRP) is a new application of tissue engineering and a developing area for clinicians and researchers. It is a storage vehicle of growth factors (GFs) such as platelet-derived growth factor (PDGF)- AA, -BB, -AB; transforming growth factor (TGF)-beta1 and -2; platelet-derived epidermal growth factor (PDEGF); platelet-derived angiogenesis factor (PDAF); insulin growth factor-1 (IGF-1); and platelet factor- 4 (PF-4), which are known to influence bone regeneration. However, animal and clinical studies reveal different results with the use of PRP and its effect on bone healing. This could be due to the differences between species, that is, differences between species in GF concentrations or variation in presence of GFs between the various PRPs. In this study, rat bone marrow cells were cultured in PRP-coated wells or in uncoated wells for 16 days in osteogenic medium, and analyzed on cell growth (DNA content) and cell differentiation (alkaline phosphatase [ALP] activity, calcium content, scanning electron microscopy, and QPCR). The concentrations of TGF-beta1, PDGF-AA, PDGF-AB, and PDGF-BB in rat, goat, and human PRP were subsequently determined. The results showed that PRP stimulated initial cell growth and had no effect on ALP activity. The calcium measurements showed a significant increase in calcium at days 8, 12, and 16. The real-time PCR results showed that PRP upregulated osteocalcin at day 1 and collagen type I at day 8. Overall, the immunoassays revealed that human PRP contained higher concentrations of growth factors per platelet compared to rat and goat PRP. Goat PRP showed higher concentrations of growth factors per platelet as compared to rat PRP except for PDGF-BB, which had a higher concentration in rat PRP. TGF-beta1 was the most abundant growth factor in all 3 PRPs. On the basis of our results, we conclude that platelet-rich plasma contains osteo-inductive growth factors, which are probably species related. However, we cannot generalize the results because of large intraspecies variations. Further, we conclude that rat PRP gel stimulates initial growth and differentiation of rat bone marrow cells in vitro.
van den Dolder
Juliette
J
Department of Periodontology and Biomaterials, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands. j.vandendolder@dent.umcn.nl
Mooren
Rob
R
Vloon
Annelies P G
AP
Stoelinga
Paul J W
PJ
Jansen
John A
JA
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D001792:Blood Platelets / Q000502:physiology*; D001854:Bone Marrow Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D002118:Calcium / Q000737:chemistry; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000378:metabolism; D024042:Collagen Type I / Q000502:physiology; D003470:Culture Media / Q000737:chemistry; D004247:DNA / Q000032:analysis; D006041:Goats; D006801:Humans; D007118:Immunoassay; D007334:Insulin-Like Growth Factor I / Q000032:analysis / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis* / Q000494:pharmacology*; D008297:Male; D015675:Osteocalcin / Q000502:physiology; D010982:Platelet-Derived Growth Factor / Q000032:analysis / Q000494:pharmacology; D053657:Platelet-Rich Plasma / Q000378:metabolism*; D020574:Proto-Oncogene Proteins c-sis; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000494:pharmacology; D053781:Transforming Growth Factor beta2 / Q000032:analysis / Q000494:pharmacology; D015854:Up-Regulation
D020099:Coated Materials, Biocompatible; D024042:Collagen Type I; D003470:Culture Media; D036341:Intercellular Signaling Peptides and Proteins; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; C095477:platelet-derived growth factor AB; D015675:Osteocalcin; D000077214:Becaplermin; D007334:Insulin-Like Growth Factor I; D004247:DNA; D000469:Alkaline Phosphatase; D002118:Calcium