Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control
Although platelet-rich plasma (PRP) is now widely used in regenerative medicine and dentistry, contradictory clinical outcomes have often been obtained. To minimize such differences and to obtain high quality evidence from clinical studies, the PRP preparation protocol needs to be standardized. In a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-06-01
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| Series: | Dentistry Journal |
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| Online Access: | https://www.mdpi.com/2304-6767/7/2/61 |
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| author | Tetsuhiro Tsujino Kazushige Isobe Hideo Kawabata Hachidai Aizawa Sadahiro Yamaguchi Yutaka Kitamura Hideo Masuki Taisuke Watanabe Hajime Okudera Koh Nakata Tomoyuki Kawase |
| author_facet | Tetsuhiro Tsujino Kazushige Isobe Hideo Kawabata Hachidai Aizawa Sadahiro Yamaguchi Yutaka Kitamura Hideo Masuki Taisuke Watanabe Hajime Okudera Koh Nakata Tomoyuki Kawase |
| author_sort | Tetsuhiro Tsujino |
| collection | DOAJ |
| description | Although platelet-rich plasma (PRP) is now widely used in regenerative medicine and dentistry, contradictory clinical outcomes have often been obtained. To minimize such differences and to obtain high quality evidence from clinical studies, the PRP preparation protocol needs to be standardized. In addition, emphasis must be placed on quality control. Following our previous spectrophotometric method of platelet counting, in this study, another simple and convenient spectrophotometric method to determine platelet aggregation activity has been developed. Citrated blood samples were collected from healthy donors and used. After centrifugation twice, platelets were suspended in phosphate buffered saline (PBS) and adenosine diphosphate (ADP)-induced aggregation was determined using a spectrophotometer at 615 nm. For validation, platelets pretreated with aspirin, an antiplatelet agent, or hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), an oxidative stress-inducing agent, were also analyzed. Optimal platelet concentration, assay buffer solution, and representative time point for determination of aggregation were found to be 50−100 × 10<sup>4</sup>/μL, PBS, and 3 min after stimulation, respectively. Suppressed or injured platelets showed a significantly lower aggregation response to ADP. Therefore, it suggests that this spectrophotometric method may be useful in quick chair-side evaluation of individual PRP quality. |
| first_indexed | 2024-04-13T09:05:11Z |
| format | Article |
| id | doaj.art-12b47fdb07384108a38e862583890644 |
| institution | Directory Open Access Journal |
| issn | 2304-6767 |
| language | English |
| last_indexed | 2024-04-13T09:05:11Z |
| publishDate | 2019-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Dentistry Journal |
| spelling | doaj.art-12b47fdb07384108a38e8625838906442022-12-22T02:53:00ZengMDPI AGDentistry Journal2304-67672019-06-01726110.3390/dj7020061dj7020061Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality ControlTetsuhiro Tsujino0Kazushige Isobe1Hideo Kawabata2Hachidai Aizawa3Sadahiro Yamaguchi4Yutaka Kitamura5Hideo Masuki6Taisuke Watanabe7Hajime Okudera8Koh Nakata9Tomoyuki Kawase10Tokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanTokyo Plastic Dental Society, Kita-ku, Tokyo 114-0002, JapanBioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata 951-8520, JapanDivision of Oral Bioengineering, Institute of Medicine and Dentistry, Niigata University, Niigata 951-8514, JapanAlthough platelet-rich plasma (PRP) is now widely used in regenerative medicine and dentistry, contradictory clinical outcomes have often been obtained. To minimize such differences and to obtain high quality evidence from clinical studies, the PRP preparation protocol needs to be standardized. In addition, emphasis must be placed on quality control. Following our previous spectrophotometric method of platelet counting, in this study, another simple and convenient spectrophotometric method to determine platelet aggregation activity has been developed. Citrated blood samples were collected from healthy donors and used. After centrifugation twice, platelets were suspended in phosphate buffered saline (PBS) and adenosine diphosphate (ADP)-induced aggregation was determined using a spectrophotometer at 615 nm. For validation, platelets pretreated with aspirin, an antiplatelet agent, or hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), an oxidative stress-inducing agent, were also analyzed. Optimal platelet concentration, assay buffer solution, and representative time point for determination of aggregation were found to be 50−100 × 10<sup>4</sup>/μL, PBS, and 3 min after stimulation, respectively. Suppressed or injured platelets showed a significantly lower aggregation response to ADP. Therefore, it suggests that this spectrophotometric method may be useful in quick chair-side evaluation of individual PRP quality.https://www.mdpi.com/2304-6767/7/2/61platelet-rich plasmaplateletsaggregationspectrophotometerquality assurance |
| spellingShingle | Tetsuhiro Tsujino Kazushige Isobe Hideo Kawabata Hachidai Aizawa Sadahiro Yamaguchi Yutaka Kitamura Hideo Masuki Taisuke Watanabe Hajime Okudera Koh Nakata Tomoyuki Kawase Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control Dentistry Journal platelet-rich plasma platelets aggregation spectrophotometer quality assurance |
| title | Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control |
| title_full | Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control |
| title_fullStr | Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control |
| title_full_unstemmed | Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control |
| title_short | Spectrophotometric Determination of the Aggregation Activity of Platelets in Platelet-Rich Plasma for Better Quality Control |
| title_sort | spectrophotometric determination of the aggregation activity of platelets in platelet rich plasma for better quality control |
| topic | platelet-rich plasma platelets aggregation spectrophotometer quality assurance |
| url | https://www.mdpi.com/2304-6767/7/2/61 |
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