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补充活化血小板分泌组的胶原屏障膜的释放动力学和促有丝分裂能力——牙周膜和成纤维细胞的体外反应

Release kinetics and mitogenic capacity of collagen barrier membranes supplemented with secretome of activated platelets - the in vitro response of fibroblasts of the periodontal ligament and the gingiva.

作者信息

Mozgan Eva-Maria, Edelmayer Michael, Janjić Klara, Pensch Manuela, Fischer Michael B, Moritz Andreas, Agis Hermann

机构信息

Department of Oral Surgery, School of Dentistry, Medical University of Vienna, Sensengasse 2a, 1090, Vienna, Austria.

Austrian Cluster for Tissue Regeneration, Donaueschingenstr. 13, 1200, Vienna, Austria.

出版信息

BMC Oral Health. 2017 Mar 21;17(1):66. doi: 10.1186/s12903-017-0357-6.

DOI:10.1186/s12903-017-0357-6
PMID:28327149
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5361806/
Abstract

BACKGROUND

Platelet preparations can stimulate the healing process and have mitogenic properties. We hypothesized that collagen barrier membranes (CBM), clinically used in guided bone regeneration and guided tissue regeneration, can serve as carriers for platelet secretome.

METHODS

Secretome was generated from washed platelets and unwashed platelets (washed/unwashed PSEC) and lyophilized onto CBM. Overall appearance of CBM was evaluated by scanning electron microscopy. The impact of PSEC on cell attachment was measured based on fluorescence microscopy with DiI-labeled cells. To assess the release kinetics, supernatants of CBM were collected and medium was replaced at hour 1-48. The mitogenic effect was evaluated with periodontal fibroblasts. Furthermore, the release of total protein, platelet-derived growth factor (PDGF)-BB, and transforming growth factor (TGF) β1 was measured.

RESULTS

CBM overall appearance and cell attachment was not modulated by PSEC. Supernatants taken after one hour induced a mitogenic response in fibroblasts and showed the highest levels of total protein, TGFβ1 and PDGF-BB. These effects decreased rapidly in subsequent supernatants. While supernatants of CBM loaded with unwashed PSEC induced a stronger mitogenic response than supernatants of CBM loaded with washed PSEC this difference between the PSEC preparations was not observed when cells were seeded on 48-hours-washed CBM.

CONCLUSIONS

CBM release platelet-derived factors in continuously declining release kinetics.

摘要

背景

血小板制剂可刺激愈合过程并具有促有丝分裂特性。我们推测，临床上用于引导骨再生和引导组织再生的胶原屏障膜（CBM）可作为血小板分泌组的载体。

方法

从洗涤血小板和未洗涤血小板（洗涤/未洗涤血小板分泌组，PSEC）中生成分泌组，并冻干到CBM上。通过扫描电子显微镜评估CBM的整体外观。基于用DiI标记细胞的荧光显微镜测量PSEC对细胞附着的影响。为评估释放动力学，收集CBM的上清液，并在第1至48小时更换培养基。用牙周成纤维细胞评估促有丝分裂作用。此外，测量总蛋白、血小板衍生生长因子（PDGF）-BB和转化生长因子（TGF）β1的释放。

结果

CBM的整体外观和细胞附着不受PSEC的调节。一小时后采集的上清液在成纤维细胞中诱导了促有丝分裂反应，并显示出总蛋白、TGFβ1和PDGF-BB的最高水平。这些作用在随后的上清液中迅速下降。虽然加载未洗涤PSEC的CBM上清液比加载洗涤PSEC的CBM上清液诱导更强的促有丝分裂反应，但当细胞接种在48小时洗涤的CBM上时，未观察到PSEC制剂之间的这种差异。

结论

CBM以持续下降的释放动力学释放血小板衍生因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/00cc856a2b43/12903_2017_357_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/5335167cd3b9/12903_2017_357_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/8e3110233261/12903_2017_357_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/ddf42229e22d/12903_2017_357_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/00cc856a2b43/12903_2017_357_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/5335167cd3b9/12903_2017_357_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/8e3110233261/12903_2017_357_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/ddf42229e22d/12903_2017_357_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/964d/5361806/00cc856a2b43/12903_2017_357_Fig4_HTML.jpg

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