2016~2017 최신 A-PRF i-PRF 관련 논문

최신 2016-1017 Low speed concept, A-PRF I-PRF 논문

Eur J Trauma Emerg Surg. 2017 Mar 21. doi: 10.1007/s00068-017-0785-7. [Epub ahead of print]

Reduction of relative centrifugal forces increases growth factor release within solid platelet-rich-fibrin (PRF)-based matrices: a proof of concept of LSCC (low speed centrifugation concept).

El Bagdadi K¹, Kubesch A¹, Yu X², Al-Maawi S¹, Orlowska A¹, Dias A¹, Booms P¹, Dohle E¹, Sader R¹, Kirkpatrick CJ¹, Choukroun J^1,3, Ghanaati S⁴.

∙FORM (Frankfurt Orofacial Regenerative Medicine) Lab, Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, University Hospital Frankfurt Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany.

∙Department of Orthopedics, West China Hospital/West China School of Medicine, Sichuan University, Chengdu, Sichuan, People's Republic of China.

∙Private Practice, Pain Therapy Center, Nice, France.

∙FORM (Frankfurt Orofacial Regenerative Medicine) Lab, Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, University Hospital Frankfurt Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany. shahram.ghanaati@kgu.de.

Abstract

Purpose The present study evaluated the platelet distribution pattern and growth factor release (VEGF, TGF-β1 and EGF) within three PRF (platelet-rich-fibrin) matrices (PRF, A-PRF and A-PRF+) that were prepared using different relative centrifugation forces (RCF) and centrifugation times. Materials and methods immunohistochemistry was conducted to assess the platelet distribution pattern within three PRF matrices. The growth factor release was measured over 10 days using ELISA. Results The VEGF protein content showed the highest release on day 7; A-PRF+ showed a significantly higher rate than A-PRF and PRF. The accumulated release on day 10 was significantly higher in A-PRF+ compared with A-PRF and PRF. TGF-β1 release in A-PRF and A-PRF+ showed significantly higher values on days 7 and 10 compared with PRF. EGF release revealed a maximum at 24 h in all groups. Toward the end of the study, A-PRF+ demonstrated significantly higher EGF release than PRF. The accumulated growth factor releases of TGF-β1 and EGF on day 10 were significantly higher in A-PRF+ and A-PRF than in PRF. Moreover, platelets were located homogenously throughout the matrix in the A-PRF and A-PRF+ groups, whereas platelets in PRF were primarily observed within the lower portion. ​Discussion the present results show an increase growthfactor release by decreased RCF. However, further studies must be conducted to examine the extent to which enhancing the amount and the rate of released growth factors influence wound healing and biomaterial-based tissue regeneration. ​Conclusion These outcomes accentuate the fact that with a reduction of RCF according to the previously LSCC (described low speed centrifugation concept), growth factor release can be increased in leukocytes and platelets within the solid PRF matrices.

Eur J Trauma Emerg Surg. 2017 Mar 10. doi: 10.1007/s00068-017-0767-9. [Epub ahead of print]

Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients' own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept.

Choukroun J^1,2, Ghanaati S³.

∙Private Practice, Pain Therapy Center, Nice, France. joseph@a-prf.com.

∙Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, FORM (Frankfurt Orofacial Regenerative Medicine) Laboratory, University Hospital Frankfurt Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany. joseph@a-prf.com.

∙Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, FORM (Frankfurt Orofacial Regenerative Medicine) Laboratory, University Hospital Frankfurt Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany. shahram.ghanaati@kgu.de

Abstract

PURPOSE:

The aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF).

MATERIALS AND METHODS:

Systematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I-III) within the spectrum (710-44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA.

RESULTS:

Reducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors' concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h.

DISCUSSION:

Based on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine.

CONCLUSIONS:

We postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.

 

Platelets. 2017 Mar 29:1-8. doi: 10.1080/09537104.2017.1293807. [Epub ahead of print]

Effects of an injectable platelet-rich fibrin on osteoblast behavior and bone tissue formation in comparison to platelet-rich plasma.

Wang X¹, Zhang Y¹, Choukroun J², Ghanaati S³, Miron RJ^4,5.

a Department of Oral Implantology , University of Wuhan , Wuhan, China.

b Pain Clinic , Nice , France.

c FORM, Frankfurt Oral Regenerative Medicine, Clinic for Maxillofacial and Plastic Surgery , Johann Wolfgang Goethe University , Frankfurt Am Main , Germany.

d Department of Periodontology, College of Dental Medicine , Nova Southeastern University , Fort Lauderdale , Florida , USA.

e Cell Therapy Institute, Centre for Collaborative Research , Nova Southeastern University , Fort Lauderdale , Florida , USA.

Abstract

Platelet-rich plasma (PRP) has been utilized for many years as a regenerative agent capable of inducing vascularization of various tissues using blood-derived growth factors. Despite this, drawbacks mostly related to the additional use of anti-coagulants found in PRP have been shown to inhibit the wound healing process. For these reasons, a novel platelet concentrate has recently been developed with no additives by utilizing lower centrifugation speeds. The purpose of this study was therefore to investigate osteoblast behavior of this novel therapy (injectable-platelet-rich fibrin; i-PRF, 100% natural with no additives) when compared to traditional PRP. Human primary osteoblasts were cultured with either i-PRF or PRP and compared to control tissue culture plastic. A live/dead assay, migration assay as well as a cell adhesion/proliferation assay were investigated. Furthermore, osteoblast differentiation was assessed by alkaline phosphatase (ALP), alizarin red and osteocalcin staining, as well as real-time PCR for genes encoding Runx2, ALP, collagen1 and osteocalcin. The results showed that all cells had high survival rates throughout the entire study period irrespective of culture-conditions. While PRP induced a significant 2-fold increase in osteoblast migration, i-PRF demonstrated a 3-fold increase in migration when compared to control tissue-culture plastic and PRP. While no differences were observed for cell attachment, i-PRF induced a significantly higher proliferation rate at three and five days when compared to PRP. Furthermore, i-PRF induced significantly greater ALP staining at 7 days and alizarin red staining at 14 days. A significant increase in mRNA levels of ALP, Runx2 and osteocalcin, as well as immunofluorescent staining of osteocalcin was also observed in the i-PRF group when compared to PRP. In conclusion, the results from the present study favored the use of the naturally-formulated i-PRF when compared to traditional PRP with anti-coagulants. Further investigation into the direct role of fibrin and leukocytes contained within i-PRF are therefore warranted to better elucidate their positive role in i-PRF on tissue wound healing.

 최신 2016-2017 A-PRF, i-PRF 논문

Int J Mol Sci. 2017 Feb 4;18(2). pii: E331. doi: 10.3390/ijms18020331.

Behavior of Gingival Fibroblasts on Titanium Implant Surfaces in Combination with either Injectable-PRF or PRP.

Wang X^1,2, Zhang Y^3,4, Choukroun J⁵, Ghanaati S⁶, Miron RJ^7,8,9.

∙The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan ∙University, Wuhan 430079, China. wangxuzhu@whu.edu.cn.

∙Department of Oral Implantology, School and Hospital of Stomatology, Wuhan University, Wuhan 430079, China. wangxuzhu@whu.edu.cn.

∙The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan ∙University, Wuhan 430079, China. zyf@whu.edu.cn.

∙Department of Oral Implantology, School and Hospital of Stomatology, Wuhan University, Wuhan 430079, China. zyf@whu.edu.cn.

∙Pain Clinic, 06000 Nice, France. joseph@a-prf.com.

∙FORM, Frankfurt Oral Regenerative Medicine, Clinic for Maxillofacial and Plastic Surgery, Johann Wolfgang Goethe University, 60596 Frankfurt Am Main, Germany. s.ghanaati@med.uni-frankfurt.de.

∙Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL 33328, USA. rmiron@nova.edu.

∙Cell Therapy Institute, Collaborative Centre for Research, Nova Southeastern University, Fort Lauderdale, FL 33328, USA. rmiron@nova.edu.

 

∙Department of Periodontics and Oral Surgery, University of Ann Arbor, Ann Arbor, MI 48109, USA. rmiron@nova.edu.

Abstract

Various strategies have been employed to speed tissue regeneration using bioactive molecules. Interestingly, platelet concentrates derived from a patient's own blood have been utilized as a regenerative strategy in recent years. In the present study, a novel liquid platelet formulation prepared without the use of anti-coagulants (injectable-platelet-rich fibrin, i-PRF) was compared to standard platelet-rich plasma (PRP) with gingival fibroblasts cultured on smooth and roughened titanium implant surfaces. Standard PRP and i-PRF (centrifuged at 700 rpm (60× g) for 3 min) were compared by assays for fibroblast biocompatibility, migration, adhesion, proliferation, as well as ｅxpression of platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), collagen1 (COL1) and fibronectin (FN). The results demonstrate that i-PRF induced significantly higher cell migration, as well as higher messenger RNA (mRNA) levels of PDGF, TGF-β, collagen1 and fibronectin when compared to PRP. Furthermore, collagen1 synthesis was highest in the i-PRF group. These findings demonstrate that liquid platelet concentrates can be formulated without the use of anticoagulants and present much translational potential for future research. Future animal and clinical trials are now necessary to further investigate the potential of utilizing i-PRF for soft tissue regenerative protocols in combination with various biomaterials.

Clin Oral Investig. 2017 Feb 2. doi: 10.1007/s00784-017-2063-9. [Epub ahead of print]

 

Injectable platelet rich fibrin (i-PRF): opportunities in regenerative dentistry?

Miron RJ^1,2,3, Fujioka-Kobayashi M^4,5,6, Hernandez M⁴, Kandalam U⁷, Zhang Y⁸, Ghanaati S⁹, Choukroun J¹⁰.

∙Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA. rmiron@nova.edu.

∙Cell Therapy Institute, Center for Collaborative Research, Nova Southeastern University, Fort Lauderdale, FL, USA. rmiron@nova.edu.

∙Department of Periodontics and Oral Medicine, University of Michigan, Ann Arbor, MI, USA. rmiron@nova.edu.

∙Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA.

∙Department of Cranio-Maxillofacial Surgery, University of Bern, Bern, Switzerland.

∙Department of Oral Surgery, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, Japan.

∙Department of Pediatric Dentistry, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA.

∙Department of Oral Implantology, University of Wuhan, Wuhan, China.

∙FORM, Frankfurt Oral Regenerative Medicine, Clinic for Maxillofacial and Plastic Surgery, Johann Wolfgang Goethe University, Frankfurt Am Main, Germany.

∙Pain Clinic, Nice, France

Abstract

OBJECTIVES:

Platelet rich plasma (PRP) has been utilized in regenerative dentistry as a supra-physiological concentrate of autologous growth factors capable of stimulating tissue regeneration. Despite this, concerns have been expressed regarding the use of anti-coagulants, agents known to inhibit wound healing. In this study, a liquid formulation of platelet rich fibrin (PRF) termed injectable-PRF (i-PRF) without the use of anti-coagulants was investigated.

MATERIALS AND METHODS:

Standard PRP and i-PRF (centrifuged at 700 rpm (60G) for 3 min) were compared for growth factor release up to 10 days (8 donor samples). Furthermore, fibroblast biocompatibility at 24 h (live/dead assay); migration at 24 h; proliferation at 1, 3, and 5 days, and ｅxpression of PDGF, TGF-β, and collagen1 at 3 and 7 days were investigated.

RESULTS:

Growth factor release demonstrated that in general PRP had higher early release of growth factors whereas i-PRF showed significantly higher levels of total long-term release of PDGF-AA, PDGF-AB, EGF, and IGF-1 after 10 days. PRP showed higher levels of TGF-β1 and VEGF at 10 days. While both formulations exhibited high biocompatibility and higher fibroblast migration and proliferation when compared to control tissue-culture plastic, i-PRF induced significantly highest migration whereas PRP demonstrated significantly highest cellular proliferation. Furthermore, i-PRF showed significantly highest mRNA levels of TGF-β at 7 days, PDGF at 3 days, and collagen1 ｅxpression at both 3 and 7 days when compared to PRP.

CONCLUSIONS:

i-PRF demonstrated the ability to release higher concentrations of various growth factors and induced higher fibroblast migration and ｅxpression of PDGF, TGF-β, and collagen1. Future animal research is now necessary to further validate the use of i-PRF as a bioactive agent capable of stimulating tissue regeneration.

CLINICAL RELEVANCE:

The findings from the present study demonstrate that a potent formulation of liquid platelet concentrates could be obtained without use of anti-coagulants.

 

 

 

 

Rev Col Bras Cir. 2015 Nov-Dec;42(6):421-3. doi: 10.1590/0100-69912015006013.

Obtention of injectable platelets rich-fibrin (i-PRF) and its polymerization with bone graft: technical note.

[Article in English, Portuguese]

Mourão CF¹, Valiense H¹, Melo ER², Mourão NB³, Maia MD¹.

∙Faculdade de Odontologia, Universidade Federal Fluminense, Rio de Janeiro, RJ, Brasil.

∙Marinha do Brasil, Rio de Janeiro, RJ, Brasil.

∙Universidade Federal Fluminense, Rio de Janeiro, RJ, Brasil.

 

Abstract

The use of autologous platelet concentrates, represent a promising and innovator tools in the medicine and dentistry today. The goal is to accelerate hard and soft tissue healing. Among them, the platelet-rich plasma (PRP) is the main alternative for use in liquid form (injectable). These injectable form of platelet concentrates are often used in regenerative procedures and demonstrate good results. The aim of this study is to present an alternative to these platelet concentrates using the platelet-rich fibrin in liquid form (injectable) and its use with particulated bone graft materials in the polymerized form.

 

Int J Implant Dent. 2016 Dec;2(1):19. Epub 2016 Aug 22.

Growth factor and pro-inflammatory cytokine contents in platelet-rich plasma (PRP), plasma rich in growth factors (PRGF), advanced platelet-rich fibrin (A-PRF), and concentrated growth factors (CGF).

Masuki H¹, Okudera T¹, Watanebe T¹, Suzuki M¹, Nishiyama K¹, Okudera H¹, Nakata K², Uematsu K³, Su CY⁴, Kawase T⁵.

∙Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan.

∙Bioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata, Japan.

∙Division of Oral Bioengineering, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan.

∙Department of Dentistry, National Yang-Ming University, Taipei, Taiwan.

∙Division of Oral Bioengineering, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan. kawase@dent.niigata-u.ac.jp.

 

Abstract

BACKGROUND:

The development of platelet-rich fibrin (PRF) drastically simplified the preparation procedure of platelet-concentrated biomaterials, such as platelet-rich plasma (PRP), and facilitated their clinical application. PRF's clinical effectiveness has often been demonstrated in pre-clinical and clinical studies; however, it is still controversial whether growth factors are significantly concentrated in PRF preparations to facilitate wound healing and tissue regeneration. To address this matter, we performed a comparative study of growth factor contents in PRP and its derivatives, such as advanced PRF (A-PRF) and concentrated growth factors (CGF).

METHODS:

PRP and its derivatives were prepared from the same peripheral blood samples collected from healthy donors. A-PRF and CGF preparations were homogenized and centrifuged to produce extracts. Platelet and white blood cell counts in A-PRF and CGF preparations were determined by subtracting those counts in red blood cell fractions, supernatant acellular serum fractions, and A-PRF/CGF exudate fractions from those counts of whole blood samples. Concentrations of growth factors (TGF-β1, PDGF-BB, VEGF) and pro-inflammatory cytokines (IL-1β, IL-6) were determined using ELISA kits.

RESULTS:

Compared to PRP preparations, both A-PRF and CGF extracts contained compatible or higher levels of platelets and platelet-derived growth factors. In a cell proliferation assay, both A-PRF and CGF extracts significantly stimulated the proliferation of human periosteal cells without significant reduction at higher doses.

CONCLUSIONS:

These data clearly demonstrate that both A-PRF and CGF preparations contain significant amounts of growth factors capable of stimulating periosteal cell proliferation, suggesting that A-PRF and CGF preparations function not only as a scaffolding material but also as a reservoir to deliver certain growth factors at the site of application.

 

J Periodontol. 2017 Jan;88(1):112-121. Epub 2016 Sep 2.

Optimized Platelet-Rich Fibrin With the Low-Speed Concept: Growth Factor Release, Biocompatibility, and Cellular Response.

Fujioka-Kobayashi M^1,2,3, Miron RJ¹, Hernandez M¹, Kandalam U⁴, Zhang Y⁵, Choukroun J⁶.

∙Department of Periodontology, College of Dental Medicine, Nova Southeastern University, Fort Lauderdale, FL.

∙Department of Cranio-Maxillofacial Surgery, University of Bern, Bern, Switzerland.

∙Department of Oral Surgery, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima, Japan.

∙Department of Pediatric Dentistry, College of Dental Medicine, Nova Southeastern University.

∙Department of Oral Implantology, University of Wuhan, Wuhan, China.

∙Pain Clinic, Nice, France.

Abstract

BACKGROUND:

Over the past decade, use of leukocyte platelet-rich fibrin (L-PRF) has gained tremendous momentum in regenerative dentistry as a low-cost fibrin matrix used for tissue regeneration. This study characterizes how centrifugation speed (G-force) along with centrifugation time influence growth factor release from fibrin clots, as well as the cellular activity of gingival fibroblasts exposed to each PRF matrix.

METHODS:

Standard L-PRF served as a control (2,700 revolutions per minute [rpm]-12 minutes). Two test groups using low-speed (1,300 rpm-14 minutes, termed advanced PRF [A-PRF]) and low-speed + time (1,300 rpm-8 minutes; A-PRF+) were investigated. Each PRF matrix was tested for growth factor release up to 10 days (eight donor samples) as well as biocompatibility and cellular activity.

RESULTS:

The low-speed concept (A-PRF, A-PRF+) demonstrated a significant increase in growth factor release of platelet-derived growth factor (PDGF), transforming growth factor (TGF)-β1, epidermal growth factor, and insulin-like growth factor, with A-PRF+ being highest of all groups. Although all PRF formulations were extremely biocompatible due to their autogenous sources, both A-PRF and A-PRF+ demonstrated significantly higher levels of human fibroblast migration and proliferation compared with L-PRF. Furthermore, gingival fibroblasts cultured with A-PRF+ demonstrated significantly higher messenger RNA (mRNA) levels of PDGF, TGF-β, and collagen1 at either 3 or 7 days.

CONCLUSIONS:

The findings from the present study demonstrate modifications to centrifugation speed and time with the low-speed concept favor an increase in growth factor release from PRF clots. This, in turn, may directly influence tissue regeneration by increasing fibroblast migration, proliferation, and collagen mRNA levels. Future animal and clinical studies are now necessary.