Fig. 3. Peri-implant bone level at baseline, 3 months post-operative, and 6 months post-operative. a Mesial defect depth. b Mesial defect width. c Distal defect depth. d Distal defect width
Fig. 3. Peri-implant bone level at baseline, 3 months post-operative, and 6 months post-operative. a Mesial defect depth. b Mesial defect width. c Distal defect depth. d Distal defect width
Fig. 2. Test group (PRF augmentation)
Fig. 2. Test group (PRF augmentation)
Fig. 1. Control group (no PRF augmentation)
Fig. 1. Control group (no PRF augmentation)
Illustration 11. ive (a), 1 week post-operative (b), 1 month post-operative (c), and 3 months post-operative (d)
Illustration 11. a–d Post-operative healing process at 3 days post-operative (a), 1 week post-operative (b), 1 month post-operative (c), and 3 months post-operative (d)
Illustration 10. a–d Second measuring, reentry, and insertion of a screwed, full ceramic crown
Illustration 10. a–d Second measuring, reentry, and insertion of a screwed, full ceramic crown
Illustration 9. a–c Radiographic control a at time of implant placement (a), 3 months post-operative (b), and 6 months post-operative (c)
Illustration 8. Fixation of the flap with Seralene® 6.0
Illustration 8. Fixation of the flap with Seralene® 6.0
Illustration 7. Insertion of PRF membranes in a double-layered technique for tissue augmentation
Illustration 7. Insertion of PRF membranes in a double-layered technique for tissue augmentation
Illustration 6. PRF membrane made by centrifugating and pressing the patient’s blood
Illustration 6. PRF membrane made by centrifugating and pressing the patient’s blood
Illustration 5. Implant placed with a split-flap technique
Illustration 5. Implant placed with a split-flap technique
Illustration 4. NobelSpeedy Replace® (source: https://www.nobelbiocare.com/de/de/home/products-and-solutions/implant-systems/nobelspeedy.html)
Illustration 4. NobelSpeedy Replace® (source: https://www.nobelbiocare.com/de/de/home/products-and-solutions/implant-systems/nobelspeedy.html)
Illustration 3. Insertion of the implant
Illustration 3. Insertion of the implant
Illustration 2. Crestal incision and preparation of a split-flap
Illustration 2. Crestal incision and preparation of a split-flap
Illustration 1. Measurement of tissue thickness with an endodontic micro-opener
Illustration 1. Measurement of tissue thickness with an endodontic micro-opener
Hehn, J., Schwenk, T., Striegel, M. et al. The effect of PRF (platelet-rich fibrin) inserted with a split-flap technique on soft tissue thickening and initial marginal bone loss around implants: results of a randomized, controlled clinical trial.
Int J Implant Dent 2, 13 (2016). https://doi.org/10.1186/s40729-016-0044-4
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Received: 30 July 2015
Accepted: 23 ...
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
Julia Hehn, Thomas Schwenk, Marcus Striegel, and Markus Schlee declare that they have no competing interests. This study was self-supported.
JH was responsible for the study design and recruitment of the patients, carried out the implant surgery and follow-up examinations, and drafted the manuscript. TS participated in the surgical treatment and follow-up examinations. MS performed the digital me...
Periodontology, Edel & Weiß Clinic, Ludwigsplatz 1a, 90403, Nuremberg, Germany
Julia Hehn
Esthetic Dentistry, Edel & Weiß Clinic, Nuremberg, Germany
Thomas Schwenk & Markus Striegel
Private Clinic for Periodontology, Forchheim and Department of Maxillofacial Surgery, Goethe University, Frankfurt, Germany
Markus Schlee
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The authors thank Dr. Wolfgang Reimers of MedCommTools for performing the statistics.
Abrahamsson I, Berglundh T. Effects of different implant surfaces and designs on marginal bone-level alterations: a review. Clin Oral Implants Res. 2009;20 Suppl 4:207–15.
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Dohan DM et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part II: platelet-related biologic features. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101(3):e45–50.
Choukroun J, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part IV: clinical effects on tissue healing. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101(3)...
Cardaropoli G et al. Tissue alterations at implant-supported single-tooth replacements: a 1-year prospective clinical study. Clin Oral Implants Res. 2006;17(2):165–71.
Fickl S et al. Dimensional changes of the alveolar ridge contour after different socket preservation techniques. J Clin Periodontol. 2008;35(10):906–13.
Vela-Nebot X et al. Benefits of an implant platform modification techniqu...
platelet-derived growth factor
platelet-rich fibrin
standard deviation
transforming growth factor-b
Soft tissue augmentation with PRF using a split-flap technique cannot be recommended to alter thin gingiva types. Future experimental and clinical studies will be necessary to evaluate whether augmentation with PRF is suitable for mucosa thickening.
With respect to marginal bone loss, it could be shown that there were no significant differences when comparing dimension of bone loss between test and control groups. The study of present research data shows, to the best knowledge of the authors, no other RCTs about tissue thickening with PRF and peri-implant bone loss. However, several studies focused on mucosa thickening with tissue grafts and ...
In this study over a period of 6 months, it could be demonstrated that mucosal tissue thickening above implants with PRF led to reduced tissue thickness when performed in a split-flap technique.
The initial post-operative dehiscence and the associated complete loss of mucosal and augmented tissue above the implant were observed in all test patients.
PRF is supposed to be a good healing aid in v...
The mean marginal bone level alterations are displayed in Fig. 3.
Six months after surgery, both groups showed a statistically significant peri-implant bone loss.
The mesial resp. distal defect depth was 0.70 mm ± 0.72 SD resp. 0.64 mm ± 0.66 SD at baseline. Six months later, the marginal bone defect increased to 1.47 mm ± 0.65 SD on the mesial side resp. 1.46 mm ± 0.54 ...
At time of surgery, the patients ranged in age from 33 to 79 years (mean age 53.8 years).
The first surgeries for implant placement in the test group were carried out as described above. Two layers of a PRF matrix were placed on top of the implant. Though surgical flaps were all sutured completely free of tension, a post-operative dehiscence above the implant could be observed in all test pati...
Comparison of bone level alterations between test and control groups.
Data were expressed as means ± standard deviation. Comparisons were made using the Wilcoxon test, the Mann–Whitney U test, and the multiple comparisons test by Schaich-Hamerle (p = 0.05).
After the implantation, standardized digital X-rays were taken with parallel technique (baseline) (illustration 9). For each patient, an individual customized digital film holder was fabricated to ensure a reproducible radiographic analysis. Patients were instructed to avoid chewing hard nutrition in the treated area and to use chlorhexidine mouthwash and a soft brush twice a day for the first 2 ...
Patients aged 18+ who required an implant in the posterior mandible were eligible for this study.
Exclusion criteria were the following:
general contraindications to implant surgery
insufficient oral hygiene and periodontitis
patients with a history of severe periodontitis
bone augmentation required
smokers
substance abuse
uncontrolled diabetes
severe cardiovascular problems
treated or u...
As a consequence, recent research focused on soft tissue augmentation of thin gingiva types prior to or simultaneous to implant insertion. Wiesner et al. published a significant gain of soft tissue by thickening the gingiva with a connective soft tissue graft harvested from the palate [12]. Soft tissues at augmented sites were 1.3 mm thicker than on control sites and had a better pink esthetic sc...
The initial bone modeling around implants within the first year after insertion presents a challenging topic in current research. Previous studies have shown that this process is characterized by a remodeling of the horizontal and vertical bone dimension with a range of 0.7 to 3 mm within the first year [1].
First attempts to reduce this loss of tissue focused on changes of implant shapes, impla...
Previous studies have shown that adequate thickness or initial augmentation of soft tissue has a positive effect on the stability of peri-implant bone. This randomized, controlled trial aimed to evaluate the influence of augmenting soft tissue with platelet-rich fibrin (PRF) on crestal bone and soft tissue around implants.
After randomization, 31 fully threaded titanium implants were inserted in ...
Fig. 4. Effects of PRP, PRGF, A-PRF, and CGF on the proliferation of human periosteal cells. Cells were treated with PRP preparations, PRGF preparations, A-PRF extracts, or CGF extracts at the indicated doses for 48 h in 1 % FBS-containing medium. *P
Fig. 3. Concentrations of the pro-inflammatory cytokines (IL-1β, IL-6) in PRP, PRGF, A-PRF, and CGF preparations (n = 20)
Fig. 3. Concentrations of the pro-inflammatory cytokines (IL-1β, IL-6) in PRP, PRGF, A-PRF, and CGF preparations (n = 20)
Preparation type
Models
Rotor
Radius
(mm)
Rotational speed
...
Masuki, H., Okudera, T., Watanebe, T. et al. 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).
Int J Implant Dent 2, 19 (2016). https://doi.org/10.1186/s40729-016-0052-4
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Received: 21 June 2016
Accepted: 18 Au...
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
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Correspondence to
Tomoyuki Kawase.
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Hideo Masuki, Toshimitsu Okudera, Taisuke Watanebe, Masashi Suzuki, Kazuhiko Nishiyama & Hajime Okudera
Bioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata, Japan
Koh Nakata
Department of Dentistry, National Yang-Ming University, Taipei, Taiwan
Chen-Yao Su
Division of Oral Bioengineering, Instit...
Kobayashi M, Kawase T, Okuda K, Wolff LF, Yoshie H. In vitro immunological and biological evaluations of the angiogenic potential of platelet-rich fibrin preparations: a standardized comparison with PRP preparations. Int J Implant Dent. 2015;1:31.
Sonnenschein SK, Meyle J. Local inflammatory reactions in patients with diabetes and periodontitis. Periodontol 2000. 2015;69:221–54.
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Clipet F, Tricot S, Alno N, Massot M, Solhi H, Cathelineau G, Perez F, De Mello G, Pellen-Mussi P. In vitro effects of Choukroun’s platelet-rich fibrin conditioned medium on 3 different cell lines implicated in dental implantology. Implant Dent. 2012;21:51–6.
Gassling VL, Acil Y, Springer IN, Hubert N, Wiltfang J. Platelet-rich plasma and platelet-rich fibrin in human cell culture. Oral Surg ...
Kawase T. Platelet-rich plasma and its derivatives as promising bioactive materials for regenerative medicine: basic principles and concepts underlying recent advances. Odontology. 2015;103:126–35.
Anitua E. The use of plasma-rich growth factors (PRGF) in oral surgery. Pract Proced Aesthet Dent. 2001;13:487–93.
Choukroun J. Advanced PRF, &i-PRF: platelet concentrates or blood concentrates? J...
Acid citrate dextrose solution
Analysis of variance
Advanced platelet-rich fibrin
A-PRF extract
Concentrated growth factors
CGF extract
Enzyme-linked immunosorbent assay
Interleukin-1β
Interleukin-6
Platelet-derived growth factor-BB
Platelet
Plasma rich in growth factors
Platelet-rich plasma
Red blood cell
Transforming growth factor-β1
Vascular endothelial growth factor
White blo...
The present study clearly demonstrated that both A-PRF and CGF preparations contained significant amounts of growth factors, which makes us to believe that A-PRF and CGF preparations would not only function as a scaffolding material but also as a reservoir to deliver certain growth factors at the site of application. Accordingly, it is expected that these two preparations are more potently capable...
In the previous study [12], we found that PRP and A-PRF preparations exert distinguishable actions on periosteal cell proliferation. Because both IL-1β and IL-6 are known to be produced by WBCs [23], and because WBCs are not included in PRGF preparations, we thought that the bi-phasic effects of PRP preparations may be attributed to WBCs. Furthermore, if WBCs are highly concentrated in A-PRF, it ...
Although the growth factor contents in PRF and CGF preparations and their bioactivities have been demonstrated in in vitro studies by several independent groups [8–11, 13–20], many clinicians still believe that the regenerative effects of PRF/CGF are solely due to fibrin clots. We speculate that this discrepancy may be caused by two major factors. First, the initial report on PRF by Choukroun ...
Numbers of platelets in PRP and PRGF preparations are shown in Fig. 1 (upper panel). Platelets were significantly concentrated both in the PRP and PRGF preparations, and the concentration rate of PRP preparations was substantially higher than that of PRGF preparations (8.79-fold vs. 2.84-fold). Numbers of platelets in A-PRF and CGF preparations calculated by the indirect subtraction method are al...
Because alveolar periosteum is closely contributed to periodontal skeletal tissue regeneration, we used human alveolar bone-derived periosteal cells for evaluation of efficacy of the PRP derivatives. The periosteal cells were obtained and expanded as described below. With informed consent, human periosteum tissue segments were aseptically dissected from the periodontal tissues of the healthy bucca...
As described previously [7, 9], blood samples (~9.5 mL) collected without anticoagulants using vacuum plain glass tubes (A-PRF+: Jiangxi Fenglin Medical Technology Co. Ltd, Fengcheng, China) or conventional vacuum plain glass tube (Plain BD Vacutainer Tube; Becton, Dickinson and Company, Franklin Lakes, NJ, USA) from the same donors were immediately centrifuged by an A-PRF centrifugation system (...
Based on their characteristics and fractionation, the differences among PRP and PRP derivatives are concisely described in our previous article [1].
As previously described [5, 6], blood samples (11.5 mL) were collected using syringes or vacuum blood collection tubes equipped with 18G needles from seven non-smoking, healthy, middle-aged, male volunteers (37 to 68 years old) three times with a 2...
Platelet-rich plasma (PRP) was originally demonstrated to be effective in the operation of alveolar ridge augmentation and immediately spread to the fields of periodontal and oral maxillofacial surgery [1]. This clinical application was endorsed by evidence that several major growth factors are contained at high levels in PRP preparations. However, for some reasons, such as low handling efficiency...
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 ...
Fig. 4. Bioactivities and PDGF-BB concentrations in PRF extracts and the supernatant serum fraction. a PRF extracts were added to periosteal cell cultures and incubated for 3 days to evaluate their effects on cell proliferation. No significant differences were observed among three groups. b PRF extracts were subjected to measurement of PDGF-BB levels using an ELISA kit. No significant differenc...
Fig. 3. SEM examination of fibrin fibers formed in self-clotted PRF and thrombin-stimulated PPP clots. PRF was prepared from fresh and 2-day-old WB samples. Similar observations were obtained from WB samples collected from three other donors. Scale bars = 10 μm
Fig. 3. SEM examination of fibrin fibers formed in self-clotted PRF and thrombin-stimulated PPP clots. PRF was prepared from fresh ...
Fig. 2. Appearance of PRF clots prepared from WB samples stored for 2 days. These observations are representative of WB samples obtained from four donors
Fig. 2. Appearance of PRF clots prepared from WB samples stored for 2 days. These observations are representative of WB samples obtained from four donors
Fig. 1. Glucose levels (a), calcium levels (b), and pH (c) of stored WB samples. Supernatant serum fractions were examined. Plasma fractions prepared by quick centrifugation were used to determine calcium levels in fresh and stored WB samples that were not added CaCl2. N = 6
Fig. 1. Glucose levels (a), calcium levels (b), and pH (c) of stored WB samples. Supernatant serum fractions were...
Isobe, K., Suzuki, M., Watanabe, T. et al. Platelet-rich fibrin prepared from stored whole-blood samples. Int J Implant Dent 3, 6 (2017). https://doi.org/10.1186/s40729-017-0068-4
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Received: 21 December 2016
Accepted: 15 February 2017
Published: 01 March 2017
DOI: https://doi.org/10.1186/s40729-017-0068-4
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
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Correspondence to Tomo...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Kazushige Isobe, Masashi Suzuki, Taisuke Watanabe, Yutaka Kitamura, Taiji Suzuki, Hideo Kawabata, Masayuki Nakamura, Toshimitsu Okudera & Hajime Okudera
Division of Oral Implantology, Niigata University Medical and Dental Hospital, Niigata, Japan
Kohya Uematsu
Bioscience Medical Research Center, Niigata University Medical and Dental ...
Duan X, Bradbury SR, Olsen BR, Berendsen AD. Matrix Biol. 2016;52-54:127–40.
Kawase T, Okuda K, Kogami H, Nakayama H, Nagata M, Nakata K, Yoshie H. Characterization of human cultured periosteal sheets expressing bone-forming potential: in vitro and in vivo animal studies. J Tissue Eng Regen Med. 2009;3:218–29.
Kawase T, Hayama K, Tsuchimochi M, Nagata M, Okuda K, Yoshie H, Burns DM, Nakata K...
Hess J. Conventional blood banking and blood component storage regulation: opportunities for improvement. Blood Transfus. 2010;8 Suppl 3:s9–s15.
World Health Organization. Manual on the management, maintenance and use of blood cold chain equipment. http://www.who.int/bloodsafety/Manual_on_Management,Maintenance_and_Use_of_Blood_Cold_Chain_Equipment.pdf. Accessed 26 Nov 2016.
van der Meer PF, d...
Acid citrate dextrose solution
Concentrated growth factors
Citrate phosphate and dextrose
Ethylenediaminetetraacetic acid
Platelet-derived growth factor
Platelet-poor plasma
Platelet-rich fibrin
Platelet-rich plasma
Scanning electron microscope
Whole blood
The self-clotted types of platelet concentrates (PRF) can be prepared from ACD-containing stored WB by addition of CaCl2 without a significant reduction in their bioactivity and without other specific reagents or devices. This approach should contribute to dissemination of PRF therapy.
Considering the current status of clinical use of platelet concentrates in the fields of periodontology and oral surgery, in this study, we used 10-mL glass tubes that are not oxygen-permeable instead of oxygen-permeable plastic bags for storage of large volumes of WB or platelets. We advanced a working hypothesis that the storage of WB samples in glass tubes would result in a more rapid and subst...
Platelet preservation is restricted to 3 and 5 days in Japan and worldwide, respectively. This limit is based on the fact that platelets are sensitive to changes in temperature and pH: when samples are stored at 2 to 6 °C, platelets become unsuitable for production of platelet concentrates [3]. Preservation of platelet concentrates results in a drop of pH below 6.0 depending on the platelet cou...
Glucose and calcium contents and pH of WB or serum samples after centrifugation are shown in Fig. 1. Because glucose is contained in the ACD-A solution, glucose levels in the stored WB and serum samples (see Fig. 4c) after centrifugation were significantly greater than those of freshly collected WB samples. Total free calcium levels, including calcium chelated by citrate, in WB samples decreased...
PRF extracts prepared as described above were subjected to measurement of PDGF-BB levels using the Human PDGF-BB Quantikine ELISA Kit (R&D Systems, Inc., Minneapolis, MN, USA) as described previously [8].
The PRF clots that were compressed in a stainless-steel compressor were fixed with 2.5% neutralized glutaraldehyde, dehydrated with a series of ethanol solutions and t-butanol, freeze-dried, and...
Stored WB samples were then mixed intermittently with 200 μL (20 μL × 10 times) of 10% CaCl2 solution and centrifuged by a Medifuge centrifugation system to prepare PRF. When lower amounts of CaCl2 were added, PRF clots were less reproducibly prepared. When higher amounts of CaCl2 were added intermittently, or when the optimal amount of CaCl2 were added at once, PRF clots were never prepa...
The study design and consent forms for all procedures performed on the study subjects were approved by the ethics committee for human subjects at Niigata University School of Medicine in accordance with the Helsinki Declaration of 1975 as revised in 2008.
With informed consent, blood samples (~9.0 mL per tube) were collected from six non-smoking, healthy, male volunteers (27 to 67 years old) us...
Blood preservation is generally and widely used in the fields of blood transfusion and surgery for either autologous or allogeneic blood [1–3]. In case of small lots of blood-derived materials used in regenerative therapy, such as platelet concentrates, it is generally accepted that autologous blood samples should be collected on-site and immediately centrifuged for processing [4]. Accordingly, ...
In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics.
Blood samples were collected ...
Fig. 3. Enzymatic degradability of A-PRF, CGF, and PPTF membranes. Each membrane disk (φ8 mm, 1 mm thick) was immersed in PBS containing trypsin and incubated in a CO2 incubator. N = 4. The asterisks represent significant differences (P
Fig. 2. Representative stress-strain curves for A-PRF and CGF membranes and mechanical properties (Young’s modulus, strain at break, and maximum stress) of A-PRF, CGF, and PPTF membranes. N = 3–9
Fig. 2. Representative stress-strain curves for A-PRF and CGF membranes and mechanical properties (Young’s modulus, strain at break, and maximum stress) of A-PRF, CGF, and PPTF membranes. N...
Fig. 1. Surface microstructures of A-PRF, CGF, and fibrin clots prepared by PPP + CaCl2 and PPTF (fibrin clots prepared by PPP and thrombin). Similar observations were obtained from other three independent blood samples. Scale bar = 10 μm. Note: the same magnification (×9000) was used in all the SEM images shown here
Fig. 1. Surface microstructures of A-PRF, CGF, and fibrin clots p...
A-PRF
CGF
PPTF
Centrifugal conditions
198 g × 8 min
692 g × 2 mina
...
Wet weight (g)
Dry weight (g)
Water content (%)
A-PRF
1.905 ± 0.416
0.043 ± 0.014*
...
Size (W × L mm)
Stretching (times longer)
Number
A-PRF
8.6 ± 1.2 × 27.5 ± 3.5
...
Isobe, K., Watanebe, T., Kawabata, H. et al. Mechanical and degradation properties of advanced platelet-rich fibrin (A-PRF), concentrated growth factors (CGF), and platelet-poor plasma-derived fibrin (PPTF).
Int J Implant Dent 3, 17 (2017). https://doi.org/10.1186/s40729-017-0081-7
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Received: 29 January 2017
Accepted: 25 April 2017
Published: 02 May 2017
...
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
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Correspondence to
Tomoyuki Kawa...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Kazushige Isobe, Taisuke Watanebe, Hideo Kawabata, Yutaka Kitamura, Toshimitsu Okudera & Hajime Okudera
Division of Dental Implantology, Niigata University Medical and Dental Hospital, Niigata, Japan
Kohya Uematsu
Division of Periodontology, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan
Kazuhiro Okuda
Bioscie...
Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost. 2005;3:1894–904.
Kawase T, Kamiya M, Kobayashi M, Tanaka T, Okuda K, Wolff LF, Yoshie H. The heat-compression technique for the conversion of platelet-rich fibrin preparation to a barrier membrane with a reduced rate of biodegradation. J Biomed Mater Res B Appl Biomater. 2015;103:825–31.
Hartshorne J, Gluckman H. A...
Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, Dohan AJ, Mouhyi J, Dohan DM. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part V: histologic evaluations of PRF effects on bone allograft maturation in sinus lift. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101:299–303.
Kawase T. Platelet-rich plasma and its derivatives as promising bioac...
Acid citrate dextrose solution
Advanced platelet-rich fibrin
Concentrated growth factors
Platelet-poor plasma-derived, thrombin-activated fibrin
Platelet-rich plasma
In the mechanical parameters and degradability we tested, CGF membranes were almost identical to A-PRF membranes. In contrast, PPTF membranes were mechanically weaker and highly degradable. Therefore, we conclude that all of these fibrin membranes are tough enough to serve as barrier membranes; however, we should pay attention to their degradability and choose an appropriate membrane type dependin...
Growth factor release is a key function of these fibrin clots for tissue regeneration. Our previous study [16] demonstrated that CGF membranes compressed by the stainless steel compression device contain significantly higher levels of growth factors even after releasing approximately 85% of exudate. Repeated rinsing with PBS failed to completely remove the growth factors from CGF membranes. The ri...
In this study, we found no apparent differences between A-PRF and CGF clot microstructures, especially in fibrin fiber thickness or crosslink density. However, in PPTF clots, which were prepared through direct conversion of fibrinogen by thrombin, fibrin fiber thickness and their crosslink density were substantially thinner and higher, respectively, than those of either A-PTF or CGF clots. This fi...
The main purpose of this study was to compare A-PRF with CGF preparations to find possible differences in mechanical properties. As shown in Table 1, the sizes of A-PRF clots compressed to membranes were 8.6 ± 1.2 mm (W) × 27.5 ± 3.5 mm (L) and very similar to those of CGF clots (8.4 ± 0.8 mm × 27.6 ± 2.5 mm). As reference, PPTF membranes were also prepared by ad...
After pipetting the digestion solution, 50 μL of the digestion solution was collected every 20 min and was stored at −20 °C until protein measurement. Protein levels, which can be considered primarily as levels of digested fibrin fiber, were then determined by a BCA protein assay kit (Takara Bio, Kusatsu, Japan). The protein levels at the time point when the initial fibrin disks were comple...
The mechanical properties of gel sheets were measured at a stretching speed of 1 mm/min with a desktop universal testing machine (EZ test; Shimadzu, Kyoto, Japan), of which maximum load cell capacity was 500 N under standard ambient conditions at 25 ± 3 °C and 50 ± 25% RH. The samples were gripped by clamps at each end (using slip-proof rubber sheets to prevent slippage) such that th...
Blood samples were collected from four non-smoking, healthy, male volunteers with ages ranging from 27 to 56 years. Although having lifestyle-related diseases and receiving medication, these donors had no hindrance in daily life. The study design and consent forms for all procedures performed with the study subjects were approved by the ethical committee for human subjects at Niigata University S...
In this study, we hypothesized that the mechanical properties of the fibrin membrane are closely related to its degradability. We compared these characteristics among A-PRF, CGF, and PPTF membranes through tensile and digestion tests.
Platelet-rich fibrin (PRF), a self-clotted preparation of platelet-concentrated, blood-derived biomaterials, is prepared solely by contact activation of intrinsic coagulation pathways through centrifugation without addition of coagulation factors [1, 2]. Therefore, the preparation protocol is drastically simplified, and the resulting clot can be handled easily with forceps. PRF is further modified...
Fibrin clot membranes prepared from advanced platelet-rich fibrin (A-PRF) or concentrated growth factors (CGF), despite their relatively rapid biodegradability, have been used as bioactive barrier membranes for alveolar bone tissue regeneration. As the membranes degrade, it is thought that the growth factors are gradually released. However, the mechanical and degradable properties of these membran...
Isobe, K., Suzuki, M., Watanabe, T. et al. Platelet-rich fibrin prepared from stored whole-blood samples.
Int J Implant Dent 3, 6 (2017). https://doi.org/10.1186/s40729-017-0068-4
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Received: 21 December 2016
Accepted: 15 February 2017
Published: 01 March 2017
DOI: https://doi.org/10.1186/s40729-017-0068-4
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
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...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Kazushige Isobe, Masashi Suzuki, Taisuke Watanabe, Yutaka Kitamura, Taiji Suzuki, Hideo Kawabata, Masayuki Nakamura, Toshimitsu Okudera & Hajime Okudera
Division of Oral Implantology, Niigata University Medical and Dental Hospital, Niigata, Japan
Kohya Uematsu
Bioscience Medical Research Center, Niigata University Medical and Dental ...
Duan X, Bradbury SR, Olsen BR, Berendsen AD. Matrix Biol. 2016;52-54:127–40.
Kawase T, Okuda K, Kogami H, Nakayama H, Nagata M, Nakata K, Yoshie H. Characterization of human cultured periosteal sheets expressing bone-forming potential: in vitro and in vivo animal studies. J Tissue Eng Regen Med. 2009;3:218–29.
Kawase T, Hayama K, Tsuchimochi M, Nagata M, Okuda K, Yoshie H, Burns DM, Nakata K...
Hess J. Conventional blood banking and blood component storage regulation: opportunities for improvement. Blood Transfus. 2010;8 Suppl 3:s9–s15.
World Health Organization. Manual on the management, maintenance and use of blood cold chain equipment. http://www.who.int/bloodsafety/Manual_on_Management,Maintenance_and_Use_of_Blood_Cold_Chain_Equipment.pdf. Accessed 26 Nov 2016.
van der Meer PF, d...
Acid citrate dextrose solution
Concentrated growth factors
Citrate phosphate and dextrose
Ethylenediaminetetraacetic acid
Platelet-derived growth factor
Platelet-poor plasma
Platelet-rich fibrin
Platelet-rich plasma
Scanning electron microscope
Whole blood
The self-clotted types of platelet concentrates (PRF) can be prepared from ACD-containing stored WB by addition of CaCl2 without a significant reduction in their bioactivity and without other specific reagents or devices. This approach should contribute to dissemination of PRF therapy.
Considering the current status of clinical use of platelet concentrates in the fields of periodontology and oral surgery, in this study, we used 10-mL glass tubes that are not oxygen-permeable instead of oxygen-permeable plastic bags for storage of large volumes of WB or platelets. We advanced a working hypothesis that the storage of WB samples in glass tubes would result in a more rapid and subst...
Platelet preservation is restricted to 3 and 5 days in Japan and worldwide, respectively. This limit is based on the fact that platelets are sensitive to changes in temperature and pH: when samples are stored at 2 to 6 °C, platelets become unsuitable for production of platelet concentrates [3]. Preservation of platelet concentrates results in a drop of pH below 6.0 depending on the platelet cou...
Glucose and calcium contents and pH of WB or serum samples after centrifugation are shown in Fig. 1. Because glucose is contained in the ACD-A solution, glucose levels in the stored WB and serum samples (see Fig. 4c) after centrifugation were significantly greater than those of freshly collected WB samples. Total free calcium levels, including calcium chelated by citrate, in WB samples decreased...
PRF extracts prepared as described above were subjected to measurement of PDGF-BB levels using the Human PDGF-BB Quantikine ELISA Kit (R&D Systems, Inc., Minneapolis, MN, USA) as described previously [8].
The PRF clots that were compressed in a stainless-steel compressor were fixed with 2.5% neutralized glutaraldehyde, dehydrated with a series of ethanol solutions and t-butanol, freeze-dried, and...
Stored WB samples were then mixed intermittently with 200 μL (20 μL × 10 times) of 10% CaCl2 solution and centrifuged by a Medifuge centrifugation system to prepare PRF. When lower amounts of CaCl2 were added, PRF clots were less reproducibly prepared. When higher amounts of CaCl2 were added intermittently, or when the optimal amount of CaCl2 were added at once, PRF clots were never prepa...
The study design and consent forms for all procedures performed on the study subjects were approved by the ethics committee for human subjects at Niigata University School of Medicine in accordance with the Helsinki Declaration of 1975 as revised in 2008.
With informed consent, blood samples (~9.0 mL per tube) were collected from six non-smoking, healthy, male volunteers (27 to 67 years old) us...
Blood preservation is generally and widely used in the fields of blood transfusion and surgery for either autologous or allogeneic blood [1–3]. In case of small lots of blood-derived materials used in regenerative therapy, such as platelet concentrates, it is generally accepted that autologous blood samples should be collected on-site and immediately centrifuged for processing [4]. Accordingly, ...
In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics.
Blood samples were collected ...
Fig. 3. Enzymatic degradability of A-PRF, CGF, and PPTF membranes. Each membrane disk (φ8 mm, 1 mm thick) was immersed in PBS containing trypsin and incubated in a CO2 incubator. N = 4. The asterisks represent significant differences (P
Fig. 2. Representative stress-strain curves for A-PRF and CGF membranes and mechanical properties (Young’s modulus, strain at break, and maximum stress) of A-PRF, CGF, and PPTF membranes. N = 3–9
Fig. 2. Representative stress-strain curves for A-PRF and CGF membranes and mechanical properties (Young’s modulus, strain at break, and maximum stress) of A-PRF, CGF, and PPTF membranes. N...
Fig. 1. Surface microstructures of A-PRF, CGF, and fibrin clots prepared by PPP + CaCl2 and PPTF (fibrin clots prepared by PPP and thrombin). Similar observations were obtained from other three independent blood samples. Scale bar = 10 μm. Note: the same magnification (×9000) was used in all the SEM images shown here
Fig. 1. Surface microstructures of A-PRF, CGF, and fibrin clots p...
A-PRF
CGF
PPTF
Centrifugal conditions
198 g × 8 min
692 g × 2 mina
...
Wet weight (g)
Dry weight (g)
Water content (%)
A-PRF
1.905 ± 0.416
0.043 ± 0.014*
...
Size (W × L mm)
Stretching (times longer)
Number
A-PRF
8.6 ± 1.2 × 27.5 ± 3.5
...
Isobe, K., Watanebe, T., Kawabata, H. et al. Mechanical and degradation properties of advanced platelet-rich fibrin (A-PRF), concentrated growth factors (CGF), and platelet-poor plasma-derived fibrin (PPTF).
Int J Implant Dent 3, 17 (2017). https://doi.org/10.1186/s40729-017-0081-7
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Received: 29 January 2017
Accepted: 25 April 2017
Published: 02 May 2017
...
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were...
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Correspondence to
Tomoyuki Kawa...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Kazushige Isobe, Taisuke Watanebe, Hideo Kawabata, Yutaka Kitamura, Toshimitsu Okudera & Hajime Okudera
Division of Dental Implantology, Niigata University Medical and Dental Hospital, Niigata, Japan
Kohya Uematsu
Division of Periodontology, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan
Kazuhiro Okuda
Bioscie...
Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost. 2005;3:1894–904.
Kawase T, Kamiya M, Kobayashi M, Tanaka T, Okuda K, Wolff LF, Yoshie H. The heat-compression technique for the conversion of platelet-rich fibrin preparation to a barrier membrane with a reduced rate of biodegradation. J Biomed Mater Res B Appl Biomater. 2015;103:825–31.
Hartshorne J, Gluckman H. A...
Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, Dohan AJ, Mouhyi J, Dohan DM. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part V: histologic evaluations of PRF effects on bone allograft maturation in sinus lift. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101:299–303.
Kawase T. Platelet-rich plasma and its derivatives as promising bioac...
Acid citrate dextrose solution
Advanced platelet-rich fibrin
Concentrated growth factors
Platelet-poor plasma-derived, thrombin-activated fibrin
Platelet-rich plasma
In the mechanical parameters and degradability we tested, CGF membranes were almost identical to A-PRF membranes. In contrast, PPTF membranes were mechanically weaker and highly degradable. Therefore, we conclude that all of these fibrin membranes are tough enough to serve as barrier membranes; however, we should pay attention to their degradability and choose an appropriate membrane type dependin...
Growth factor release is a key function of these fibrin clots for tissue regeneration. Our previous study [16] demonstrated that CGF membranes compressed by the stainless steel compression device contain significantly higher levels of growth factors even after releasing approximately 85% of exudate. Repeated rinsing with PBS failed to completely remove the growth factors from CGF membranes. The ri...
In this study, we found no apparent differences between A-PRF and CGF clot microstructures, especially in fibrin fiber thickness or crosslink density. However, in PPTF clots, which were prepared through direct conversion of fibrinogen by thrombin, fibrin fiber thickness and their crosslink density were substantially thinner and higher, respectively, than those of either A-PTF or CGF clots. This fi...
The main purpose of this study was to compare A-PRF with CGF preparations to find possible differences in mechanical properties. As shown in Table 1, the sizes of A-PRF clots compressed to membranes were 8.6 ± 1.2 mm (W) × 27.5 ± 3.5 mm (L) and very similar to those of CGF clots (8.4 ± 0.8 mm × 27.6 ± 2.5 mm). As reference, PPTF membranes were also prepared by ad...
After pipetting the digestion solution, 50 μL of the digestion solution was collected every 20 min and was stored at −20 °C until protein measurement. Protein levels, which can be considered primarily as levels of digested fibrin fiber, were then determined by a BCA protein assay kit (Takara Bio, Kusatsu, Japan). The protein levels at the time point when the initial fibrin disks were comple...
The mechanical properties of gel sheets were measured at a stretching speed of 1 mm/min with a desktop universal testing machine (EZ test; Shimadzu, Kyoto, Japan), of which maximum load cell capacity was 500 N under standard ambient conditions at 25 ± 3 °C and 50 ± 25% RH. The samples were gripped by clamps at each end (using slip-proof rubber sheets to prevent slippage) such that th...
Blood samples were collected from four non-smoking, healthy, male volunteers with ages ranging from 27 to 56 years. Although having lifestyle-related diseases and receiving medication, these donors had no hindrance in daily life. The study design and consent forms for all procedures performed with the study subjects were approved by the ethical committee for human subjects at Niigata University S...
In this study, we hypothesized that the mechanical properties of the fibrin membrane are closely related to its degradability. We compared these characteristics among A-PRF, CGF, and PPTF membranes through tensile and digestion tests.
Platelet-rich fibrin (PRF), a self-clotted preparation of platelet-concentrated, blood-derived biomaterials, is prepared solely by contact activation of intrinsic coagulation pathways through centrifugation without addition of coagulation factors [1, 2]. Therefore, the preparation protocol is drastically simplified, and the resulting clot can be handled easily with forceps. PRF is further modified...
Fibrin clot membranes prepared from advanced platelet-rich fibrin (A-PRF) or concentrated growth factors (CGF), despite their relatively rapid biodegradability, have been used as bioactive barrier membranes for alveolar bone tissue regeneration. As the membranes degrade, it is thought that the growth factors are gradually released. However, the mechanical and degradable properties of these membran...
Toyoda, T., Isobe, K., Tsujino, T. et al. Direct activation of platelets by addition of CaCl2 leads coagulation of platelet-rich plasma.
Int J Implant Dent 4, 23 (2018). https://doi.org/10.1186/s40729-018-0134-6
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Received: 13 March 2018
Accepted: 11 May 2018
Published: 01 August 2018
DOI: https://doi.org/10.1186/s40729-018-0134-6
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were ...
The study design and consent forms of all the procedures performed were approved by the ethics committee for human participants of the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013. Written informed consents to participate in the study were obtained from all the participants.
Written informed consent for the publicati...
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TT, KI, TT, and TK conceived and designed the study, performed the experiments and data analysis, and wrote the manuscript. YK, FO, TW, ...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Toshihisa Toyoda, Kazushige Isobe, Tetsuhiro Tsujino, Yasuo Koyata, Fumitaka Ohyagi, Taisuke Watanabe, Masayuki Nakamura, Yutaka Kitamura & Hajime Okudera
Bioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata, Japan
Koh Nakata
Division of Oral Bioengineering, Institute of Medicine and Dentistry, ...
The authors appreciate Shoshin EM (Okazaki, Japan) for giving an opportunity to use DHM.
Because an article performed and prepared in parallel is now submitted elsewhere for publication, the authors do not wish to share their data at present time.
Blitz A. Pump thrombosis—a riddle wrapped in a mystery inside an enigma. Ann Cardiothor Surg. 2014;3:450–71.
Kawase T, Watanabe T, Okuda K. Platelet-rich plasma and its derived platelet concentrates: what dentists involved in cell-based regenerative therapy should know. Nihon Shishubyou Gakkai Kaishi. 2017;59:68–76. (in Japanese)
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Kobayashi M, Kawase T, Horimizu M, Okuda K, Wolff LF, Yoshie H. A proposed protocol for the standardized preparation of PRF membranes for clinical use. Biologicals. 2012;40:323–9.
Kawase T, Okuda K, Nagata M, Tsuchimochi M, Yoshie H, Nakata K. Non-invasive, quantitative assessment of the morphology of gamma-irradiated human mesenchymal stem cells and periosteal cells using digital holographic m...
Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85:638–46.
Anitua E. Plasma rich in growth factors: preliminary results of use in the preparation of future sites for implants. Int J Oral Maxillofac Implants. 1999;14:529–35.
Masuki H, Okudera T, Wa...
Acid citrate dextrose solution
Adenosine diphosphate
Calcium-sensing receptor
Digital holographic microscopy
Phosphate-buffered saline
Platelet-poor plasma
Platelet-rich fibrin
Platelet-rich plasma
Scanning electron microscope
Tris-buffered saline
Thromboxane A2
In addition to the well-known intrinsic coagulation pathway, which activates platelets via thrombin conversion, added Ca2+ may directly activate washed platelets and promote clot formation alone and in cooperation with the coagulation pathway as illustrated in Fig. 8. Therefore, it is possible to control platelet activation levels and the consequent growth factor release by modulating not only in...
On the other hand, in PRF prepared from fresh whole-blood without anticoagulants, it is thought that the centrifugal force increases the contact of factor XII with a glass surface, thereby primarily forming fibrin clots. Although platelets can also be activated by a glass surface, the centrifugal force accumulates platelets at and just below the interface between the red blood cell fraction and pl...
An alternative possibility may be related to the status of platelets stored in a low-Ca2+ environment: platelets constantly increase thromboxane A2 (TXA2) production and tend to easily aggregate under the influence of the TXA2 autocrine loop [19]. Therefore, it is thought that platelets prepared from citrated whole-blood samples continuously repeat Ca2+ discharge from intracellular Ca2+ stores and...
It is well known that platelets are activated by adenosine diphosphate (ADP), thrombin, epinephrine, thromboxane A2, collagen, and many other compounds and thus aggregate through binding of fibrinogen and glycoprotein IIb/IIIa receptors and upregulate surface antigens known as “platelet activation markers”: CD62P and CD63 [13, 14]. During activation, platelet morphology generally changes from ...
The surface microstructures of the formed fibrin clots are depicted in Fig. 6. The loop-like substances that initially formed in PRP on watch glasses were composed of abundant aggregated platelets, and relatively smaller amounts of fibrin fibers were deposited around platelet aggregates (Fig. 6a). By contrast, in the final version of fibrin clots, platelet aggregates were hardly detectable and m...
Morphological changes of (and microparticle release and fibrin formation by) Ca2+-stimulated platelets are shown in Fig. 1. When washed platelets were treated with 0.1% CaCl2 in sample tubes and plated in culture dishes for the last 5 min of treatment (control, 2 min), platelets’ ability to adhere to the dish bottom surface increased with the duration of Ca2+ treatment (Fig. 1a). On the othe...
Imaging by DHM (HoloMonitor M4; Phase Holographic Imaging AB, Lund, Sweden) was performed as described elsewhere [12]. The data were analyzed using specialized software, HoloStudio M4 (Phase Holographic Imaging AB). For surface roughness and area analysis, after a series of images were captured, the grayscale images were converted to the black-and-white format by the Otsu method, and the cell iden...
As described in the subsection above, the platelet fractions were prepared and treated with CaCl2 in polypropylene sample tubes. At the end of incubation, the platelets were fixed with an equal volume of a commercial fixative, ThromboFix (Beckman-Coulter, Brea, CA, USA), for 30 min, washed twice with PBS, and probed simultaneously with phycoerythrin (PE)-conjugated mouse monoclonal anti-CD41 and ...
Blood samples were collected from eight nonsmoking healthy male volunteers at ages from 32 to 68 years. The study design and consent forms of all the procedures performed were approved by the ethics committee for human participants of the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013.
Peripheral blood (~ 9 mL) was...
In this study, we attempted to dissociate platelets from the coagulation pathway and to evaluate possible direct action of Ca2+ on platelet functions in citrated whole-blood samples. In addition, in response to recent increasingly frequent requests for scheduled or outsourced, but not immediate on-site, preparation of various platelet concentrate types [4, 5], we examined time course changes in pl...
Since Marx’s report [1], platelet-rich plasma (PRP) and subsequently modified PRP derivatives have been widely applied in regenerative dentistry. Unlike self-clotted platelet-rich fibrin (PRF), for better handling efficiency and minimizing the loss of growth factors to diffusion, PRP and some other derivatives in liquid form are usually clotted by addition of exogenous coagulation factors, such ...
Based on the notion that full activation of platelets is required for a growth factor release, in regenerative dentistry, platelet-rich plasma (PRP) in liquid form is usually clotted by addition of CaCl2 in glassware before topical implantation. However, there has been no evidence as to which is better, full or partial activation of platelets, for minimizing the loss of growth factors and improvin...
Fig. 7. Scatter plots representing correlations between measured and calculated platelet counts in P-PRP and L-PRP preparations. Note: a strong correlation was observed only in P-PRP. N = 32 and 50 for P-PRP and L-PRP, respectively
Fig. 7. Scatter plots representing correlations between measured and calculated platelet counts in P-PRP and L-PRP preparations. Note: a strong correlation was ...
Fig. 6. Scatter plots representing possible correlations between platelet (PLT) and WBC counts and between platelet and RBC counts in P-PRP and L-PRP preparations. Note: strong positive correlations were observed between platelets and RBC in both PRP types. N = 32 and 50 for P-PRP and L-PRP, respectively
Fig. 6. Scatter plots representing possible correlations between platelet (PLT) and WB...
Fig. 5. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for validation testing. N = 32 and 50 for P-PRP and L-PRP, respectively
Fig. 5. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for validation testing. N = 32 and 50 for P-PRP and L-PRP, respectively
Fig. 4. Calibration curves of measured platelet counts versus absorbance in P-PRP and L-PRP preparations. The samples were serially diluted by PPP, and the platelet counts were determined using an AHA and SPM. N = 14 for each type of PRP
Fig. 4. Calibration curves of measured platelet counts versus absorbance in P-PRP and L-PRP preparations. The samples were serially diluted by PPP, and th...
Fig. 3. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for calibration curves. N = 14 for each type of PRP
Fig. 3. Counts of platelets (PLT), WBCs, and RBCs in P-PRP and L-PRP preparations prepared for calibration curves. N = 14 for each type of PRP
Fig. 2. The appearance of blood sampled after gravity fractionation and the resulting P-PRP and L-PRP. In the first low-speed spin, samples were centrifuged for 10 min at 533×g. For P-PRP preparation, the upper plasma fraction, which was 2 mm beyond the interface between plasma and RBC fractions, was transferred into sample tubes for the second high-speed spin (2656×g, 5 min). In contrast, ...
Fig. 1. A compact SPM with its remote controller installed on an iPad Air. iPhones and other Android devices can be used instead of the iPad Air
Fig. 1. A compact SPM with its remote controller installed on an iPad Air. iPhones and other Android devices can be used instead of the iPad Air
Kitamura, Y., Suzuki, M., Tsukioka, T. et al. Spectrophotometric determination of platelet counts in platelet-rich plasma.
Int J Implant Dent 4, 29 (2018). https://doi.org/10.1186/s40729-018-0140-8
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Received: 13 April 2018
Accepted: 29 June 2018
Published: 02 October 2018
DOI: https://doi.org/10.1186/s40729-018-0140-8
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were m...
The study design and consent forms of all the procedures performed were approved by the ethics committee for human participants of the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013. Written informed consents to participate in the study were obtained from all the participants.
Written informed consents for the publicat...
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YK, MS, TyTo, and TK conceived and designed the study, performed the experiments and data analysis, and wrote the manuscript. KI, TaTn, ...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Yutaka Kitamura, Masashi Suzuki, Tsuneyuki Tsukioka, Kazushige Isobe, Tetsuhiro Tsujino, Taisuke Watanabe, Takao Watanabe & Hajime Okudera
Bioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata, Japan
Koh Nakata
Department of Materials Science and Technology, Niigata University, Niigata, Japan
Tak...
Because an article performed and prepared in parallel is now submitted elsewhere for publication, the authors do not wish to share their data at present time.
Xu Z, Yin W, Zhang Y, Qi X, Chen Y, Xie X, Zhang C. Comparative evaluation of leukocyte- and platelet-rich plasma and pure platelet-rich plasma for cartilage regeneration. Sci Rep. 2017;7:43301.
Yin W, Qi X, Zhang Y, Sheng J, Xu Z, Tao S, Xie X, Li X, Zhang C. Advantages of pure platelet-rich plasma compared with leukocyte- and platelet-rich plasma in promoting repair of bone defects. J Transl Me...
Guder WG, da Fonseca-Wollheim F, Heil W, Schmitt YM, Töpfer G, Wisser H, Zawta B. The Haemolytic, Icteric and Lipemic Sample Recommendations Regarding their Recognition and Prevention of Clinically Relevant Interferences. Recommendations of the Working Group on Preanalytical Variables of the German Society for Clinical Chemistry and the German Society for Laboratory Medicine. LaboratoriumsMedizin...
Hazan R, Que YA, Maura D, Rahme LG. A method for high throughput determination of viable bacteria cell counts in 96-well plates. BMC Microbiol. 2012;12:259.
Lee VS, Tarassenko L. An optical method for the determination of platelet count in platelet samples contaminated with red blood cells. J Biochem Biophys Methods. 1992;24:215–23.
Davis VL, Abukabda AB, Radio NM, Witt-Enderby PA, Clafshenkel...
Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: Growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85:638–46.
Etulain J. Platelets in wound healing and regenerative medicine. Platelets. 2018:1–13.
Hou X, Yuan J, Aisaiti A, Liu Y, Zhao J. The effect of platelet-rich plasma on clinical outcomes of t...
Acid-citrate-dextrose solution
Automated hematology analyzer
Leukocyte-rich PRP
Prostaglandin E1
Platelet-poor plasma
Platelet-rich fibrin
Platelet-rich plasma
Pure-PRP
Red blood cell
Standard deviation
Spectrophotometer
Leukocyte
In normal blood samples composed of light yellow plasma, spectrophotometric determination of platelet counts would be useful for quality assurance of individual PRP preparations. For accurate determination, however, operators should handle samples with care to minimize the inclusion of WBCs and RBCs in PRP preparations.
Consistent with the clinical significance of platelet counting, several studies have reported that the platelet concentration is the most reliable criterion for the regenerative ability of PRP [27, 28] because platelets increase the number of anabolic signaling molecules. Conversely, as WBCs increase the number of catabolic signaling molecules, the quality of PRP can, perhaps, be considerably alte...
Another limitation is the color of plasma. In terms of color, blood samples obtained from the donors participating in this study were light yellow and could be evaluated as “normal.” However, we have sometimes encountered colored plasma samples in clinical practice. For example, when blood triglyceride levels are high, the plasma turns milky white or turbid [22,23,24]. Hemolytic plasma looks r...
Since determination of bacterial cell number is a fundamental procedure in the field of microbiology, several methods have been developed and widely employed depending on the purpose of cell counting. SPM is one of the common methods used to estimate bacterial load [12]. The advantage of SPM is speed and convenience without additional preparation steps. On the other hand, the limitations are the i...
For validation of these calibration curves, P-PRP and L-PRP preparations prepared by three independent operators were employed. Blood cell counts are shown in Fig. 5. As observed in the calibration curves for the samples, significant differences were found in WBC and RBC counts, but not in platelet counts, between the P-PRP and L-PRP preparations. Correlations between platelet counts and WBC or R...
The appearance of the blood-collection tube after the first low-speed spin and representative P-PRP and L-PRP preparations after the second high-speed spin and subsequent re-suspension are shown in Fig. 2. Although low-speed spinning did not result in the formation of a clear buffy coat in the interface between the plasma and RBC fractions, the buffy coat corresponding to the plasma was not inclu...
Using the data obtained with both the AHA and SPM, scattered plots were created to examine correlations and obtain formulas to calculate platelet counts.
P-PRP and L-PRP preparations were independently prepared from the 11 donors by three well-trained operators. Platelet counts were first determined using the AHA and aliquots of the PRP preparations were measured using the SPM. Platelet counts we...
Blood samples were collected from 11 non-smoking healthy male volunteers aged 33 to 69 years. The study design and consent forms for all the procedures were approved by the ethics committee for human participants at the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013.
Peripheral blood (~ 9 mL) was collected into pla...
In this study, we focused on the possibility of spectrophotometric determination and validated the applicability of the proposed method on platelet counts in PRP preparations. This idea was based on bacterial cell counting [12] and a similar challenge was reported in 1992 [13]. However, this optical method has not been further modified for PRP as a grafting material for regenerative therapy in acc...
Almost two decades have passed since platelet concentrates, such as platelet-rich plasma (PRP), were first introduced to the field of regenerative medicine by Marx et al. [1]. To date, PRP has been modified to create different variations and has increasingly been used in various fields of regenerative therapy around the world. However, negative data obtained from clinical applications of PRP have ...
Platelet-rich plasma (PRP) is widely used in regenerative dentistry and other medical fields. However, its effectiveness has often been questioned. For better evaluation, the quality of individual PRP preparations should be assured prior to use. We proposed a spectrophotometric method for determination of platelet counts and validated its applicability using two types of PRP preparations.
Blood s...
Figure 3. Data from the VAS of patient-related outcome measures at the time of mounting of the implant-supported crown and at the final follow-up of the PRF and control group
Figure 2. Box plot of the radiographic peri-implant marginal bone level at different time points in millimeter. Baseline: the time of implant placement; abutment: the time of abutment operation; impression: the time of impression taking; follow-up: the time of the final follow-up
Figure 1. Intraoperative photos illustrating bone harvesting and lateral bone augmentation in the PRF group. Initially, an incision is made at the lateral aspect of the posterior part of the mandibular corpus (a) followed by exposing the mucoperiosteal flap (b), before making the osteotomy line (c). The bone block (d) is then retrieved before adjusted to the contour at the recipient site and...
Table 4 Patient-related outcome measures at baseline and at the final follow-up
Test group
Control group
Difference
p value
Mean (95% CI)
Mean (95% CI)
Mean (95% CI)
Baseline
9.44 (9.09 to 9.78)
9.57 (9.20 to 9.95)
0.13 (− 0.40 to 0.66)
0.61
Follow-up
9.66 (9.30 to 10.02)
9.55 (9.15 to 9.96)
− 0.10 (-0.66 to 0.46)
0.71
Difference
0.22 (...
Table 3 Radiographic marginal bone level and clinical recession on neighbouring tooth surface
Group
Baseline (mean, 95% CI)
Follow-up (mean, 95% CI)
Difference (mean, 95% CI)
p value
Radiographic marginal bone level in mm
Test
1.94 (1.50 to 2.38)
2.07 (1.64 to 2.51)
− 0.14 (− 0.25 to − 0.02)
p = 0.03
Control
2.34 (1.62 to 3.08)
2.49 (1.73...
Table 2 Radiographic peri-implant marginal bone level in mm
Test group
Control group
Mean difference
95% CI
p value
Obs
Mean
95% CI
Obs
Mean
95% CI
Baseline
14
− 0.24
− 0.48 to 0.00
13
− 0.28
− 0.52 to 0.03
0.04
− 0.314 to 0.39
p = 0.82
Abutment
14
0.07
− 0.17 to 0.30
13
− 0.01
− 0.26 to 0.25
0.08
− 0.278...
Table 1 Demographics and survival rates of implants and implant crowns
Test group (PRF)
Control group
Number of implants
14
13
Mean age, years (range)
47.9 (23–66)
52.3 (24–72)
Gender
Female
6
6
Male
8
7
Smokers
Total
2
1
20 cigarettes per day
1
1
Number of implants
14
13
Implant length (mm) and imp...
Abbreviations
ASC:
Angulated screw channel
BOP:
Bleeding on probing
DBBM:
Deproteinised bovine bone mineral
GBR:
Guided bone regeneration
ICC:
Intraclass correlation coefficient
KT:
Keratinised peri-implant tissue
PCR:
Plaque control record
PD:
Probing depth
PRF:
Platelet-rich fibrin
PROM:
Patient-related outcome measures
RCF:
...
A minor, but statistically significant, radiographic bone loss occurred from baseline to the final follow-up at the neighbouring tooth surfaces in both groups. Moreover, both groups experienced a minor recession of the marginal gingiva from baseline to the final follow-up, but the change was not significant. Recession and the bone level of the neighbouring tooth surfaces to implants placed in ...
The combination of the NobelParallel CC implant launched in 2015 and an abutment with ASC is relatively new and has so far been lined to only few mechanical problems [39, 40], among which rotation of the crown when torqueing the abutment screw was not stated. In both patients, a new implant was placed without any need for additional bone augmentation and without further complications. ...
Discussion
The present study focused on clinical and radiographic characteristics of staged implants placed in autogenous bone grafts covered by either a PRF membrane (PRF group) or a standard procedure (gold standard) involving coverage of the autogenous bone graft using a deproteinised bovine bone mineral and a resorbable collagen membrane (control group).
The PRF group demonstrated a hi...
One patient (control group) expressed minimally changed extraoral sensation in the chin region at both the 1- and 2-week follow-up. However, the extra- and intraoral clinical examination revealed no sensory disturbances. The patient was not affected by this and described the same changed sensation at the final clinical follow-up after 29 months.
Another patient (PRF group) experienced sensory d...
Radiographic peri-implant marginal bone change
The mean peri-implant marginal bone level at the different time points is shown in Table 2 and Fig. 2. The mean marginal bone level at follow-up was 0.26 mm (95% CI: 0.01–0.50 mm) in the PRF group and 0.68 mm (95% CI: 0.41–0.96 mm) in the control group. The difference between the groups was − 0.43 mm (95% CI: − 0.80 to − 0....
Bleeding on probing
The estimated probability or observed proportion of BOP for implants was 0.31 (95% CI: 0.14–0.70) in the PRF group and 0.30 (95% CI: 0.12–0.77) in the control group. The ratio of the probability of observing BOP was 1.046 (95% CI: 0.91–1.20), indicating that the probability of observing BOP is 4.6% higher in the PRF group than in the control group. No statistical differe...
Results
Implant survival
Two of the 27 initially placed implants were lost in the control group (Table 1). Twenty months after placement of the implant-supported crown, one implant (first premolar, regular platform (4.3 mm), length: 13 mm) was lost due to failed osseointegration. No periodontitis or peri-implant marginal bone resorption was obvious at the time of implant removal. A second...
The distance from the implant-abutment connection to the peri-implant marginal bone level was measured mesially and distally in parallel with the long axis of the implant using open-source software (ImageJ, National Institutes of Health, Bethesda, MD, USA). The distance from the cemento-enamel junction to the marginal bone level at the neighbouring tooth surfaces was also measured in parallel with...
Prosthodontic treatment
Forty-nine days (range: 27–113 days) after placement of the healing abutment, the abutment was removed and the implant position was registered by an impression coping on the implant.
The final implant-supported restoration was fabricated by using an individually designed angulated screw channel (ASC) zirconium abutment (Nobel Biocare®, Zürich, Switzerland) and venee...
Methylprednisolone was prescribed the following morning (16 mg) and evening (16 mg). Additionally, postoperative ibuprofen (400 mg, four times daily) and paracetamol (1000 mg, four times daily) were prescribed for 1 week. The patients were instructed to rinse with 0.12% chlorhexidine digluconate twice daily and discontinue the use of their prostheses (if any). Patients were seen for ...
The bone graft was retrieved by making a continuous osteotomy line with a cylindrical and a round bur at the lateral part of the mandible, with a uniform size of approximately 15 × 25 mm (Fig. 1c, d). The bone block containing mainly cortical bone was then gently separated from the mandible using a raspartorium. The block graft was covered with a saline-moistened gauze until used. In the PRF g...
At the time of the bone augmentation procedure, two patients (14%) in the PRF group and one (8%) patient in the control group were smokers. Patients were partially edentulous due to trauma (n = 22), agenesis (n = 3) or marginal periodontitis [2]. Two patients were unavailable for the final follow-up. The referring dentist followed the non-attenders, and telephone interview revealed no subjective o...
Material and methods
The study was performed according to the Declaration of Helsinki and internationally accepted guidelines for RCT, including the CONSORT statement (www.consort-statement.org). The volumetric changes of the augmented bone [26], the histological composition of the augmented bone [27] and pain after the primary bone augmentation procedure [28] were previously described in d...
Despite the shape of a membrane, the PRF membrane does not have the properties of a resorbable barrier membrane [21, 22], due to its fast degradation in the same manner as a natural blood clot (1–2 weeks) [23]. Therefore, the PRF membrane is not believed to replace a barrier membrane in the classic understanding of guide bone regeneration (GBR), but rather to enhance the healing capacity of t...
Background
Implant-supported single crowns are characterised by high long-term survival and few biological and technical complications, which typically includes peri-implant marginal bone loss, screw-loosening and fracture of veneering material complications [1,2,3]. To achieve a successful treatment outcome, the implants must be inserted in sufficient bone volume of an adequate quality to obta...
A clinical and radiographic study of implants placed in autogenous bone grafts covered by either a platelet-rich fibrin membrane or deproteinised bovine bone mineral and a collagen membrane: a pilot randomised controlled clinical trial with a 2-year follow-up
Abstract
Purpose
To compare the survival and clinical performance of implants placed in sites previously augmented with autogenous ...
Wedharan
Konsep-konsep rekayasa jaringan yang kompleks harus dievaluasi sehubungan dengan penerapan klinisnya. Dengan demikian, tujuan keseluruhan hendaklah untuk menetapkan metode yang idealnya dapat diselesaikan dalam rentang kala yang singkat sebelum atau selama prosedur bedah regeneratif sineja. Selama beberapa tahun pungkasan, PRF Choukroun sudah terbukti jadi metode yang mendekati konsep id...
Weton
Penelitian histokimia (H&E, Mason-Goldner, and Giemsa)
S-PRF
Dalam bagian longitudinal gumpalan S-PRF yang diproduksi menurut protokol sentrifugasi standar (2700 rpm, 12 menit), gumpalan fibrin padat terlihat dengan ruang interfibrosa minimal. Dengan metode pewarnaan histokimia standar, sel teramati di seluruh gumpalan tapi jumlahnya menurun menuju bagian distal dari gumpalan PRF (data ti...
Produksi PRF Choukroun
Perancah PRF disiapkan sesuai protokol yang dipublikasikan sebelumnya.11,26 Empat sukarelawan saras (yaitu, tanpa sejarah penggunaan antikoagulan) dalam rentang yuswa antara 18 sampai 60 tahun ikut serta dalam penelitian ini. Untuk setiap peserta penelitian, 4 tabung darah perifer dikumpulkan dan segera ditempatkan dalam centrifuge yang telah diprogram sebelumnya (PC-O2, PR...
Tujuan utama penelitian biomaterial dan rekayasa jaringan adalah untuk mempromosikan reaksi jaringan yang diinduksi oleh bahan dan yang mengarah ke regenerasi dan proses penyembuhan luka yang efektif di derah yang rusak. Dengan demikian, biomaterial harus berfungsi sebagai penghalang sementara untuk menutupi cacat dan meningkatkan regenerasi jaringan seraya kompatibel dengan jaringan dan, yang pal...
Fibrin Kaya Platelet Lanjutan: Konsep baru untuk rekayasa jaringan berbasis sel dengan sarana sel inflamasi
Shahram Ghanaati, MD, DMD; Patrick Booms, PhD; Anna Orlowska, BSc, DVM; Alica Kubesch; Jonas Lorenz, DDS; Jim Rutkowski, DMD, PhD; Constantin Landes, MD, DMD, PhD; Robert Sader, MD, DDS, PhD; CJ Kirkpatrick, MD, PhD, DSc; Joseph Choukroun, MD
J Oral Implant...
Discussion
Complex tissue engineering concepts have to be evaluated in terms of their clinical applicability. Thus, the overall goal should be to establish a method that could ideally be completed within a short time span before or during intended regenerative surgical procedures. Over the last few years, Choukroun's PRF has proven to be a method that comes close to the ideal concept of guide...
Results
Histochemical studies (H&E, Mason-Goldner, and Giemsa)
S-PRF
In the longitudinal section of the S-PRF clot, produced according to the standard centrifugation protocol (2700 rpm, 12 minutes), a dense fibrin clot was seen with minimal interfibrous space. With the standard histochemical staining methods, cells were observed throughout the clot, albeit decreasing toward the more distal ...
Production of Choukroun's PRF
The PRF scaffolds were prepared according to a previously published protocol.11,26 Four healthy (ie, with no history of anticoagulant usage) volunteers in an age range between 18 to 60 years participated in this study. For each individual, 4 tubes of peripheral blood were collected and immediately placed in a preprogrammed centrifuge (PC-O2, PROCESS for PRF, N...
A major objective of biomaterial research and tissue engineering is to promote a material-induced tissue reaction that leads to regeneration and an effective wound-healing process in the defective area. Thus, a biomaterial should serve as a temporary barrier to cover defects and promote tissue regeneration while being tissue compatible and, most importantly, clinically applicable. In the field o...
Advanced Platelet-Rich Fibrin: A New Concept for Cell-Based Tissue Engineering by Means of Inflammatory Cells
Shahram Ghanaati, MD, DMD; Patrick Booms, PhD; Anna Orlowska, BSc, DVM; Alica Kubesch; Jonas Lorenz, DDS; Jim Rutkowski, DMD, PhD; Constantin Landes, MD, DMD, PhD; Robert Sader, MD, DDS, PhD; CJ Kirkpatrick, MD, PhD, DSc; Joseph Choukroun, MD
J Oral Impla...