Allogeneic Fibrin Clot for Odontogenic/Cementogenic Differentiation of Human Dental Mesenchymal Stem Cells
Allogeneic Fibrin Clot for Odontogenic/Cementogenic Differentiation of Human Dental Mesenchymal Stem Cells
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DOI：
10.1007/s13770-020-00279-z
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发表时间：
2020-06-26
影响因子：
3.6
通讯作者：
Choung, Pill-Hoon
中科院分区：
文献类型：
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作者：
Cao, Hua-Lian;Chung, Jong Hoon;Choung, Pill-Hoon
背景技术背景：胎牛血清被广泛用作细胞培养基的生长补充剂;然而，动物源性病原体增加了传播传染性病原体的风险。富含血小板的纤维蛋白最近被认为是一种成功的替代品，但白细胞对其同种异体可行性存在限制。本研究旨在探讨不含白细胞的同种异体纤维蛋白凝块（AFC）在体外和体内对人牙髓干细胞（hDPSCs）和人牙周膜干细胞（hPDLSCs）向牙本质/牙骨质分化的诱导作用。方法：采用高速离心法制备AFC，去除大部分白细胞，经3次冻融循环后获得AFC血清。用AFC血清处理hDPSC和hPDLSC，通过实时聚合酶链反应研究牙本质或牙骨质形成相关标志物。用AFC血清处理hDPSCs，将其置于牙本质管内，用AFC血清处理hPDLSCs，将其包裹于牙本质外，然后将混合物移植到裸鼠皮下，持续12周。结果：AFC血清具有足够的生长因子和细胞因子诱导hDPSCs和hPDLSCs向牙本质/牙骨质分化。此外，AFC seurum可诱导hDPSCs分化为成牙本质细胞样细胞和牙髓样组织，hPDLSCs再生为牙骨质样组织。结论：AFC可作为一种安全的人牙源性间充质干细胞（hDMSCs）的来源。
BACKGROUND: Fetal bovine serum is widely used as a growth supplement for cell culture medium; however, animal-borne pathogens increase the risk of transmitting infectious agents. Platelet-rich fibrin is recently considered as a successful alternative but leukocytes present limits to its allogeneic feasibility. The aim of this study was to explore the effects of allogeneic fibrin clot (AFC) without leukocytes on inducing odontogenic/cementogenic differentiation of human dental pulp stem cells (hDPSCs) and human periodontal ligament stem cells (hPDLSCs)in vitroandin vivo. METHODS: AFC was prepared by high-speed centrifugation and leukocytes were almost removed, and AFC serum was obtained through three freeze-thaw cycles. hDPSCs and hPDLSCs were treated with AFC serum to investigate the odontogenic or cementogenic associated markers by real-time polymerase chain reaction. hDPSCs were treated with AFC serum and placed inside of dentin canal, hPDLSCs were treated with AFC serum to wrap outside of dentin, the mixture was then transplanted into the subcutaneous of nude mice for 12 weeks. RESULTS: AFC serum exhibited enough growth factors and cytokines to induce odontogenic/cementogenic differentiation of hDPSCs and hPDLSCsin vitro. Furthermore, AFC seurum could induce hDPSCs to differentiate into odontoblasts-like cells and pulp-like tissues, and hPDLSCs to regenerate cementum-like tissues. CONCLUSION: AFC could be an alternative safe source with growth factors for the expansion of human dental mesenchymal stem cells (hDMSCs).