Pulp Fibroblasts Synthesize Functional Complement Proteins Involved in Initiating Dentin-Pulp Regeneration
Pulp Fibroblasts Synthesize Functional Complement Proteins Involved in Initiating Dentin-Pulp Regeneration
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DOI：
10.1016/j.ajpath.2014.04.003
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发表时间：
2014-07-01
影响因子：
6
通讯作者：
About, Imad
中科院分区：
文献类型：
--
作者：
Chmilewsky, Fanny;Jeanneau, Charlotte;About, Imad
补体系统是控制组织损伤和感染的一种有效的血浆免疫监测系统。虽然肝脏是主要的循环补体蛋白合成部位，但众所周知，肝外合成可以优化局部组织的炎症反应。由于已知牙本质-牙髓再生受局部调控，我们研究了牙髓内局部补体系统的激活及其在启动再生过程中的作用。膜攻击复合体(C5b-9)的形成和革兰氏染色表明补体的激活与人类龋齿中革兰氏阳性细菌的存在有关。RT-PCR分析表明，在脂磷壁酸的刺激下，培养的人牙髓成纤维细胞能产生有效激活补体所需的所有蛋白质。在没有血浆蛋白的情况下，C5b-9的形成和C5a活性片段的产生在体外证明了这一点。最后，在Mu-Slide趋化小室中进行的动态迁移分析和使用C5aR特异性拮抗剂(W54011)表明，牙髓成纤维细胞合成的补体蛋白的激活和随后C5a的释放特异性地诱导了牙髓前体细胞的募集。我们的研究表明，人牙髓成纤维细胞是第一种能够合成所有补体蛋白的非免疫细胞类型。这些成纤维细胞通过补体激活来招募牙髓前体细胞，从而显著促进组织再生，这可能是一种潜在的靶向牙髓成纤维细胞的治疗策略。
The complement system is an efficient plasma immune surveillance system that controls tissue injury and infection. Although the liver constitutes the primary circulating complement protein synthesis site, extrahepatic synthesis is known to optimize local tissue inflammatory reaction. Because dentin-pulp regeneration is known to be regulated locally, we investigated activation of the local complement system within the dental pulp and its role in initiating the regeneration process. Membrane attack complex (C5b-9) formation and Gram's staining revealed that complement activation is correlated with the presence of Gram-positive bacteria in carious human teeth. RT-PCR analysis demonstrated that cultured human pulp fibroblasts stimulated with lipoteichoic acid produce all the proteins required for efficient complement activation. This was demonstrated in vitro by C5b-9 formation and C5a active fragment production in the absence of plasma proteins. Finally, the dynamic migration assays performed in mu-Slide chemotaxis chambers and use of a C5aR-specific antagonist (W54011) demonstrated that the activation of complement proteins synthesized by pulp fibroblasts and the subsequent release of C5a specifically induced pulp progenitor cell recruitment. Our study reveals human pulp fibroblasts as the first nonimmune cell type capable of synthesizing all complement proteins. These fibroblasts cells contribute significantly to tissue regeneration by recruiting pulp progenitors via complement activation, which suggests to a potential therapeutic strategy of targeting pulp fibroblasts in dentin-pulp regeneration.