Spectrophotometric determination of platelet counts in platelet-rich plasma

International Journal of Implant Dentistry, Oct 2018

Background 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. Methods Blood samples were obtained from healthy male volunteers and pure PRP (P-PRP) and leukocytes-rich PRP (L-PRP) were prepared using the double-spin method. In serial dilutions, platelet counts in P-PRP and L-PRP were determined using an automated hematology analyzer and a compact spectrophotometer. For validation, P-PRP and L-PRP independently prepared by three well-trained operators were used for comparison of the calculated and measured platelet counts. Results In the two types of PRP samples evaluated, platelet counts were almost equal and greater amount of both white blood cells (WBCs) and red blood cells (RBCs) were included in L-PRP preparations. The calibration curve obtained from serially diluted P-PRP showed a strong correlation (R2 = 0.995), whereas that of L-PRP was relatively weaker (R2 = 0.975). In validation testing, the scatter plot of the calculated platelet counts versus the measured values showed a strong correlation in P-PRP (R2 = 0.671), whereas that of L-PRP showed a much weaker correlation (R2 = 0.0605). Conclusions This method can precisely determine platelet counts in PRP preparations when the inclusion of WBCs or RBCs is minimized. Therefore, we recommend that clinicians use this method for quality assurance of individual PRP preparations.

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Spectrophotometric determination of platelet counts in platelet-rich plasma

International Journal of Implant Dentistry December 2018, 4:29 | Cite as Spectrophotometric determination of platelet counts in platelet-rich plasma AuthorsAuthors and affiliations Yutaka KitamuraMasashi SuzukiTsuneyuki TsukiokaKazushige IsobeTetsuhiro TsujinoTaisuke WatanabeTakao WatanabeHajime OkuderaKoh NakataTakaaki TanakaTomoyuki Kawase Open Access Research First Online: 02 October 2018 401 Downloads Abstract Background 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. Methods Blood samples were obtained from healthy male volunteers and pure PRP (P-PRP) and leukocytes-rich PRP (L-PRP) were prepared using the double-spin method. In serial dilutions, platelet counts in P-PRP and L-PRP were determined using an automated hematology analyzer and a compact spectrophotometer. For validation, P-PRP and L-PRP independently prepared by three well-trained operators were used for comparison of the calculated and measured platelet counts. Results In the two types of PRP samples evaluated, platelet counts were almost equal and greater amount of both white blood cells (WBCs) and red blood cells (RBCs) were included in L-PRP preparations. The calibration curve obtained from serially diluted P-PRP showed a strong correlation (R2 = 0.995), whereas that of L-PRP was relatively weaker (R2 = 0.975). In validation testing, the scatter plot of the calculated platelet counts versus the measured values showed a strong correlation in P-PRP (R2 = 0.671), whereas that of L-PRP showed a much weaker correlation (R2 = 0.0605). Conclusions This method can precisely determine platelet counts in PRP preparations when the inclusion of WBCs or RBCs is minimized. Therefore, we recommend that clinicians use this method for quality assurance of individual PRP preparations. KeywordsPlatelet Count Spectrophotometry Leukocytes Red blood cells Quality assurance  Abbreviations ACD Acid-citrate-dextrose solution AHA Automated hematology analyzer L-PRP Leukocyte-rich PRP PGE1 Prostaglandin E1 PPP Platelet-poor plasma PRF Platelet-rich fibrin PRP Platelet-rich plasma P-PRP Pure-PRP RBC Red blood cell SD Standard deviation SPM Spectrophotometer WBC Leukocyte Background 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 often been reported, leading to controversy regarding the predictability of PRP therapy [2]. Especially in cases of skeletal regeneration, the efficacy of PRP has been controversial [3, 4, 5, 6, 7, 8, 9]. One possible major reason behind this debate is the lack of large controlled clinical trials [2] or randomized clinical trials. Because there is no consensus regarding the indications and contraindications for PRP therapy, it is theoretically difficult to design appropriate experiments. In addition, there are no generally accepted guidelines on how to evaluate the condition of application sites. The second major reason, which has frequently been used as a possible explanation (actually, an “excuse”) for unexpected clinical results in many clinical case reports, is individual difference. This is highly conceivable, but not convincingly supported by scientific evidence in individual cases. The third major reason is the lack of consensus regarding PRP preparation protocols [2]. Recent advances in the development of various automated preparation devices and kits are expected to reduce not only the labor of the operator but also technique-dependent variation of PRP quality. However, it should be noted that these devices cannot standardize PRP quality. In other words, it is not guaranteed that the quality of individual PRP preparations depends specifically on individual preparation devices. In fact, it is well-known that PRP and its derivatives prepared using the same devices do not necessarily induce similar clinical results. In Japan, a new regulatory framework for PRP therapy was established in 2014. However, no evaluation indexes for PRP quality, except for aseptic handling to ensure sterility, are indicated in the regulations. In our recent review article [10], we highlighted the necessity of PRP quality indexes. The primary index is platelet counts. Specifically, it is best to check platelet counts prior to use. To assess PRP quality in clotted PRP derivatives, such as platelet-rich fibrin (PRF), we recently develope (...truncated)


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Yutaka Kitamura, Masashi Suzuki, Tsuneyuki Tsukioka, Kazushige Isobe, Tetsuhiro Tsujino, Taisuke Watanabe, Takao Watanabe, Hajime Okudera, Koh Nakata, Takaaki Tanaka, Tomoyuki Kawase. Spectrophotometric determination of platelet counts in platelet-rich plasma, International Journal of Implant Dentistry, 2018, pp. 29, Volume 4, Issue 1, DOI: 10.1186/s40729-018-0140-8