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血小板浓缩物释放的自体生长因子对牙周膜成纤维细胞成骨分化的影响:一项比较研究。

Effect of autogenous growth factors released from platelet concentrates on the osteogenic differentiation of periodontal ligament fibroblasts: a comparative study.

作者信息

Zhang Zheng, Li Xinyue, Zhao Jing, Jia Wenjun, Wang Zuomin

机构信息

Department of Periodontology, Tianjin Stomatological Hospital, Hospital of Stomatology, Nankai University, Tianjin, China.

Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.

出版信息

PeerJ. 2019 Oct 31;7:e7984. doi: 10.7717/peerj.7984. eCollection 2019.

Abstract

BACKGROUND

Platelet concentrates have been used in tissue regeneration. The purpose of this study was to examine effects of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF) and concentrated growth factor (CGF) on the osteogenic differentiation of periodontal ligament fibroblasts (PDLFs).

METHODS

Leukocyte- and platelet-rich fibrins, CGFs and PDLFs were obtained from New Zealand rabbits. The release of basic fibroblast growth factor (bFGF), bone morphogenetic protein 2 (BMP-2) and transforming growth factor β1 (TGF-β1) from L-PRFs and CGFs was measured at 5 h and 1, 3, 5, 7 days, using the enzyme linked immunosorbent assay. The PDLFs were treated with exudates of L-PRF or CGF. After the treatment, cell counting kit-8 assay was performed at day 1, 3, 5 and 7. Alkaline phosphatase (ALP) assay and Western blotting were applied at day 7. Three blocking antibodies were used to neutralize the proteins of bFGF, BMP-2 and TGF-β1.

RESULTS

Leukocyte- and platelet-rich fibrin and CGF showed different growth factor release pattern, but similar accumulated concentration of these growth factors. PDLFs proliferation was significantly promoted by both L-PRF and CGF at day 1, 3 and 7, and CGF group was superior to L-PRF group at day 1 and 3. Both L-PRF and CGF significantly enhanced PDLFs ALP activity and protein expression of osteogenic markers. The osteopontin level was higher in CGF group than in L-PRF group, but no significant differences were found between two groups for ALP activity. Three blocking antibodies significantly downregulated both L-PRF and CGF induced osteogenic markers expression.

CONCLUSION

Both CGF and L-PRF can promote the proliferation and osteogenic differentiation of PDLFs. The bFGF, BMP-2 and TGF-β1 are involved in both L-PRF and CGF induced osteogenic differentiation of PDLFs.

摘要

背景

血小板浓缩物已用于组织再生。本研究的目的是检测富含白细胞和血小板的纤维蛋白(L-PRF)及浓缩生长因子(CGF)释放的生长因子对牙周膜成纤维细胞(PDLFs)成骨分化的影响。

方法

从新西兰兔获取富含白细胞和血小板的纤维蛋白、CGF及PDLFs。采用酶联免疫吸附测定法在5小时以及第1、3、5、7天测量L-PRF和CGF中碱性成纤维细胞生长因子(bFGF)、骨形态发生蛋白2(BMP-2)和转化生长因子β1(TGF-β1)的释放量。用L-PRF或CGF的渗出液处理PDLFs。处理后,在第1、3、5和7天进行细胞计数试剂盒-8检测。在第7天进行碱性磷酸酶(ALP)检测和蛋白质印迹分析。使用三种阻断抗体中和bFGF、BMP-2和TGF-β1的蛋白质。

结果

富含白细胞和血小板的纤维蛋白和CGF呈现不同的生长因子释放模式,但这些生长因子的累积浓度相似。在第1、3和7天,L-PRF和CGF均显著促进PDLFs增殖,且在第1和3天CGF组优于L-PRF组。L-PRF和CGF均显著增强PDLFs的ALP活性和成骨标志物的蛋白质表达。CGF组骨桥蛋白水平高于L-PRF组,但两组间ALP活性无显著差异。三种阻断抗体均显著下调L-PRF和CGF诱导的成骨标志物表达。

结论

CGF和L-PRF均可促进PDLFs的增殖和成骨分化。bFGF、BMP-2和TGF-β1参与L-PRF和CGF诱导的PDLFs成骨分化过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f31/6825745/69a95490f066/peerj-07-7984-g001.jpg

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