Platelet‐Poor and Platelet‐Rich Plasma Stimulate Bone Lineage Differentiation in Periodontal Ligament Stem Cells

Martínez, Constanza; González, Sergio; Palma, Verónica; Ph, Smith

doi:10.1902/jop.2015.150360

Cited by 29 publications

(27 citation statements)

References 46 publications

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“…Such aprotocol results in a three phase-composed suspension,from which the PRF comprises the middle fraction and the red blood cells the lower portion ( 16 ). The supernatant, known as PPP, comprises the upper fraction, which is discarded after the PRF has been obtained, despitethe literature reporting that the PPP possessesa growth factor profile similar to PRF ( 13 , 14 ). In this context, this study was the first one to obtain the PRF used as natural scaffold and the use of PPP as nutritional supplement to cells culture in PRF in a single protocol.…”

Section: Discussionmentioning

confidence: 99%

“…During blood centrifugation, in order to obtainPRF, platelets are activated with a significant growth factor release - Transforming Growth Factor Beta I (TGF-β1), Insulin-Like Growth Factor I (IGF-I), Platelet-Derived Growth Factor Beta (PDGF-β) - which are sequestered in the fibrin network during the polymerization process. Recent studies ( 13 , 14 ) have shown that the supernatant resulting from the obtainedPRF, the Platelet-Poor Plasma (PPP), could contain some grow factors in its composition. In addition, the PPP is able to induce the osteoblastic differentiation of stem cells from periodontal ligament in vitro ( 14 ) and the repair of bone defects in vivo ( 13 ).…”

Section: Introductionmentioning

confidence: 99%

“…Recent studies ( 13 , 14 ) have shown that the supernatant resulting from the obtainedPRF, the Platelet-Poor Plasma (PPP), could contain some grow factors in its composition. In addition, the PPP is able to induce the osteoblastic differentiation of stem cells from periodontal ligament in vitro ( 14 ) and the repair of bone defects in vivo ( 13 ). To the best of our knowledge, there have been no reports evaluating PPPas DMEM supplementationas an alternative to FBS.…”

Section: Introductionmentioning

confidence: 99%

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Platelet-Poor Plasma as a Supplement for Fibroblasts Cultured in Platelet-Rich Fibrin

Chisini¹,

Karam²,

Noronha³

et al. 2017

Acta Stomatol Croat

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The aim of this study was to evaluate the proliferation and adhesion of mesenchymal cells (3T3/NIH) in Dulbecco’s Modified Eagle Medium(DMEM) supplemented with Platelet-Poor Plasma (PPP) in aPlatelet-Rich Fibrin (PRF) scaffold. Human blood was obtained and processed in a centrifuge considering the equation G=1.12xRx(RPM/1000)2 to obtain PRF and PPP.Cell adhesion and maintenance analyses were performed by MTTassays in a 96 well plate withsupplemented DMEM: PPP (90:10) for 24 hours. Besides, the PRF was deposited in a 48 well plate and 10x104 cells were seeded above each PRF (n=3) with 800µl of DMEM: PPP (90:10) and cultured for 7 days. Histological analysis and the immunohistochemical staining for Vimentin were performed. Results were analyzed by one-way ANOVA in Stata12®. A significant decrease (p<0.05) of cells adhesion in relationship to FBSwas observed. However, a similar ability of cell-maintenance for PPP 10% was observed (P>0.05). Fibroblasts culture for 7 days in PRF supplemented with PPP 10% was possible, showing positive staining for Vimentin. Therefore, PPP cell supplementation decreased the initial adhesion of cells but was able to maintain the proliferation of adhered cells and able to support their viability in PRF.It seems that this method has many clinical advantagessince it provides an autologous and natural scaffold with their respective supplement for cell culture by only one process, without using xenogeneic compounds. This could improve the potential of clinical translational therapies based on the use of PRF cultured cells, promoting the regenerative potential for future use in medicine and dentistry.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Platelet-Poor Plasma as a Supplement for Fibroblasts Cultured in Platelet-Rich Fibrin

Chisini¹,

Karam²,

Noronha³

et al. 2017

Acta Stomatol Croat

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“…There are many possible ways to activate PRP, which entails inducing the release of bioactive molecules from platelets and the cleavage of fibrinogen. Most commonly, thrombin and/or calcium chloride are used for this purpose [7]. Once activated, PRP will eventually form a hydrogel, which has been reported to secret growth factors [8] and may be applied as delivery vehicle for cells [8,9], as is discussed below.…”

Section: Introductionmentioning

confidence: 99%

Platelet-Rich Plasma in Tissue Engineering: Hype and Hope

Lang

Loibl²,

Herrmann³

2018

Eur Surg Res

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Background: Platelet-rich plasma (PRP) refers to an enriched platelet suspension in plasma. In addition to the clinical application of PRP in the context of various orthopedic diseases and beyond, PRP and platelet lysate (PL) have been in focus in the field of tissue engineering. In this review, we discuss the application of PRP as a cell culture supplement and as part of tissue engineering strategies, particularly emphasizing current hurdles and ambiguities regarding the efficacy of PRP in these approaches. Summary: As a putative autologous replacement for animal-derived supplements such as fetal calf serum (FCS), PRP has been applied as cell culture supplement for the expansion of stem and progenitor cells for tissue engineering applications and cell therapies. Attributed to the high content of growth factors in platelets, PRP has been shown to promote cell growth, which was mostly superior to standard cultures supplemented with FCS, while the differentiation capacity of progenitor cells seems not to be affected. However, it was also suggested that cultivation of cells with PRP significantly alters the protein expression profile in cells in comparison to FCS, indicating that the influence of PRP on cell behavior should be thoroughly investigated. Moreover, different PRP preparation methods and donor variations have to be considered for the use of PRP under good manufacturing practice conditions. PRP has been used for various tissue engineering applications in the context of bone, cartilage, skin, and soft tissue repair, where most studies were conducted in the field of bone tissue engineering. These approaches take either advantage of the release of chemoattractive, angiogenic, proliferative, and putatively pro-regenerative growth factors from PRP, and/or the hydrogel properties of activated PRP, making it suitable as a cell delivery vehicle. In many of these studies, PRP is combined with biomaterials, cells, and in some cases recombinant growth factors. Although the experimental design often does not allow conclusions on the pro-regenerative effect of PRP itself, most publications report beneficial effects if PRP is added to the tissue-engineered construct. Furthermore, it was demonstrated that the release of growth factors from PRP may be tailored and controlled when PRP is combined with materials able to capture growth factors. Key Messages: Platelet-derived preparations such as PRP and PL represent a promising source of autologous growth factors, which may be applied as cell culture supplement or to promote regeneration in tissue-engineered constructs. Furthermore, activated PRP is a promising candidate as an autologous cell carrier. However, the studies investigating PRP in these contexts often show conflicting results, which most likely can be attributed to the lack of standardized preparation methods, particularly with regard to the platelet content and donor variation of PRP. Ultimately, the use of PRP has to be tailored for the individual application.

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“…A positive effect of PPP at low concentration (2%) on myogenic differentiation of C2C12 myoblasts was observed in our study. Observations provided by Martínez et al (2016) suggest a positive effect of PPP on bone differentiation of human periodontal ligament stem cells even at low concentrations (2.5%) of this platelet-derived fraction. Recently published results by Miroshnychenko et al (2017) indicate the in vitro dose range of PPP (i.e.…”

Section: Discussionmentioning

confidence: 92%

Platelet-poor plasma of athletes is a potent inducer of myogenic differentiation of C2C12 myoblasts

et al. 2020

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Introduction. Blood products, i.e. platelet rich plasma (PRP), leukocyte-poor plasma (PRP) and platelet poor plasma (PPP), have previously been used to improve muscle regeneration. In this study, six months? frozen-stored PPP of individuals who practiced different types of physical exercise was analysed; it could steer mouse C2C12 myoblast cells towards proliferation, migration and myogenic differentiation, and it could affect the morphology/shape of myotubes. Materials and Methods. PPP of male Olympic weightlifters, football players and professional folk dancers, aged 15-19, was collected 12 h post-training and stored for 6 months at -20?C. C2C12 cell proliferation was assessed by MTT test, motility by scratch assay, myogenic differentiation by myotube formation and gelatinase activity by gel-zymography. Results and Conclusions. PPP induced proliferation and migration of C2C12 cells. Proliferative capacity was as follows: weightlifters > dancers > football players; mean migratory capacity was: weightlifters = dancers > football players. PPP induced formation of myotubes; significant inter-individual variations were detected: PPP from weightlifters induced formation of round myotubes, and PPP from football players and dancers induced formation of elongated myotubes. The mean myotube area was as follows: football players > dancers > weightlifters. PPP gelatinolytic activity was observed; it was negatively correlated with C2C12 myoblast proliferation. These results provide general but distinct evidence that PPP of individuals practicing certain types of exercise can specifically modify myoblast morphology/function. This is significant for explaining physiological responses and adaptations to exercise. In conclusion, long-term, frozen-stored PPP preserves its potential to modify myoblast morphology and function. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 175062]

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Platelet‐Poor and Platelet‐Rich Plasma Stimulate Bone Lineage Differentiation in Periodontal Ligament Stem Cells

Abstract: PRP and PPP showed a similar protein profile and exerted comparable effects on bone differentiation. Further studies are needed to characterize and compare the effects of PPP and PRP on bone healing in vivo.

Cited by 29 publications

References 46 publications

Platelet-Poor Plasma as a Supplement for Fibroblasts Cultured in Platelet-Rich Fibrin

Platelet-Poor Plasma as a Supplement for Fibroblasts Cultured in Platelet-Rich Fibrin

Platelet-Rich Plasma in Tissue Engineering: Hype and Hope

Platelet-poor plasma of athletes is a potent inducer of myogenic differentiation of C2C12 myoblasts

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