Platelet-Rich Fibrin Extract: A Promising Fetal Bovine Serum Alternative in Explant Cultures of Human Periosteal Sheets for Regenerative Therapy
Platelet-Rich Fibrin Extract: A Promising Fetal Bovine Serum Alternative in Explant Cultures of Human Periosteal Sheets for Regenerative Therapy
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DOI：
10.3390/ijms20051053
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发表时间：
2019-03-01
影响因子：
5.6
通讯作者：
Nakata, Koh
中科院分区：
文献类型：
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作者：
Kawase, Tomoyuki;Nagata, Masaki;Nakata, Koh
2004年，我们开发了用于治疗牙周骨缺损的自体骨膜片。这种再生疗法已经成功地再生了牙周骨，并增加了牙槽骨，用于种植。然而，6周培养的必要性是一个限制。在这里，我们研究了富含人血小板的纤维蛋白提取物(PRFext)作为胎牛血清(FBS)的替代品在一种最初为维持间充质干细胞而开发的新型培养液(MSC-PCM)中用于骨膜片外植体培养的适用性。将小段骨膜在MSC-PCM+2%PRFext中扩增4周，并与传统方法制备的骨膜片进行生长速度、细胞多层形成、碱性磷酸酶(ALP)活性和表面抗原(CD73、CD90、CD105)表达的比较。骨膜片在新型培养基中的生长速度快于常规培养基中。然而，细胞形态和碱性磷酸酶活性的评估表明，在新的培养液中生长的骨膜细胞相对不成熟。这些发现表明，含有PRFext的新型培养液具有缩短培养周期和排除与异种因子相关的可能的风险而不会对骨膜活性产生负面影响的优点。
In 2004, we developed autologous periosteal sheets for the treatment of periodontal bone defects. This regenerative therapy has successfully regenerated periodontal bone and augmented alveolar ridge for implant placement. However, the necessity for 6-week culture is a limitation. Here, we examined the applicability of a human platelet-rich fibrin extract (PRFext) as an alternative to fetal bovine serum (FBS) for the explant culture of periosteal sheets in a novel culture medium (MSC-PCM) originally developed for maintaining mesenchymal stem cells. Small periosteum tissue segments were expanded in MSC-PCM + 2% PRFext for 4 weeks, and the resulting periosteal sheets were compared with those prepared by the conventional method using Medium199 + 10% FBS for their growth rate, cell multilayer formation, alkaline phosphatase (ALP) activity, and surface antigen expression (CD73, CD90, and CD105). Periosteal sheets grew faster in the novel culture medium than in the conventional medium. However, assessment of cell shape and ALP activity revealed that the periosteal cells growing in the novel medium were relatively immature. These findings suggest that the novel culture medium featuring PRFext offers advantages by shortening the culture period and excluding possible risks associated with xeno-factors without negatively altering the activity of periosteal sheets.