Isobe Kazushige, Suzuki Masashi, Watanabe Taisuke, Kitamura Yutaka, Suzuki Taiji, Kawabata Hideo, Nakamura Masayuki, Okudera Toshimitsu, Okudera Hajime, Uematsu Kohya, Nakata Koh, Tanaka Takaaki, Kawase Tomoyuki
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan.
Division of Oral Implantology, Niigata University Medical and Dental Hospital, Niigata, Japan.
Int J Implant Dent. 2017 Dec;3(1):6. doi: 10.1186/s40729-017-0068-4. Epub 2017 Mar 1.
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 from non-smoking, healthy male donors (aged 27-67 years, N = 6), and PRF clots were prepared immediately or after storage for 1-2 days. Fibrin fiber was examined by scanning electron microscopy. Bioactivity was evaluated by means of a bioassay system involving human periosteal cells, whereas PDGF-BB concentrations were determined by an enzyme-linked immunosorbent assay.
Addition of optimal amounts of a 10% CaCl solution restored the coagulative ability of whole-blood samples that contained an anticoagulant (acid citrate dextrose) and were stored for up to 2 days at ambient temperature. In PRF clots prepared from the stored whole-blood samples, the thickness and cross-links of fibrin fibers were almost identical to those of freshly prepared PRF clots. PDGF-BB concentrations in the PRF extract were significantly lower in stored whole-blood samples than in fresh samples; however, both extracts had similar stimulatory effects on periosteal-cell proliferation.
Quality of PRF clots prepared from stored whole-blood samples is not reduced significantly and can be ensured for use in regenerative therapy. Therefore, the proposed method enables a more flexible treatment schedule and choice of a more suitable platelet concentrate immediately before treatment, not after blood collection.
在再生治疗中，自体凝血血小板浓缩物，如富血小板纤维蛋白（PRF），通常在现场制备并立即用于治疗。如果血液样本或制备好的凝块能够保存数天，其临床适用性将会扩大。在此，我们从储存的全血样本中制备了PRF，并检测了其特性。
从不吸烟的健康男性供者（年龄27 - 67岁，N = 6）采集血液样本，立即或储存1 - 2天后制备PRF凝块。通过扫描电子显微镜检查纤维蛋白纤维。通过涉及人骨膜细胞的生物测定系统评估生物活性，而血小板衍生生长因子 - BB（PDGF - BB）浓度通过酶联免疫吸附测定法测定。
添加适量的10%氯化钙溶液可恢复含有抗凝剂（酸性枸橼酸盐葡萄糖）并在室温下储存长达2天的全血样本的凝血能力。在由储存的全血样本制备的PRF凝块中，纤维蛋白纤维的厚度和交联与新鲜制备的PRF凝块几乎相同。储存的全血样本中PRF提取物中的PDGF - BB浓度显著低于新鲜样本；然而，两种提取物对骨膜细胞增殖具有相似的刺激作用。
从储存的全血样本制备的PRF凝块质量没有显著降低，可确保用于再生治疗。因此，所提出的方法能够实现更灵活的治疗方案，并能在治疗前而非采血后立即选择更合适的血小板浓缩物。
