Effects of platelet-rich and -poor plasma on the reparative response of gingival fibroblasts
Effects of platelet-rich and -poor plasma on the reparative response of gingival fibroblasts
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DOI：
10.1111/j.1600-0501.2011.02274.x
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发表时间：
2012-09-01
影响因子：
4.3
通讯作者：
Smith, Patricio C.
中科院分区：
文献类型：
--
作者：
Caceres, Monica;Martinez, Constanza;Smith, Patricio C.
目的富血小板血浆（PRP）作为一种促进创伤修复的治疗手段，其作用机制尚不完全清楚。本研究旨在表征PRP和贫血小板血浆（PPP）上清液对牙龈组织修复所涉及的细胞反应的影响。方法研究PRP和PPP组分对人牙龈成纤维细胞（HGF）基质收缩、细胞迁移、成肌纤维细胞分化、基质成分产生和蛋白水解酶的影响。使用商业试剂盒从供体获得PRP和PPP。通过在基质金属蛋白酶（MMP）和肌动蛋白聚合抑制剂存在下的胶原晶格评价基质收缩。通过Western-blot评估基质分子和蛋白酶的产生。通过下拉测定评价RhoA活性。通过免疫荧光法评估肌动蛋白分布和局灶性粘连。通过ELISA定量转化生长因子-β（TGF-β）。结果PRP和PPP均能刺激人牙龈成纤维细胞胶原凝胶收缩，而伊洛马司他和细胞松弛素D可抑制这一反应。PRP和PPP还刺激MT 1-MMP和TIMP-2的产生，RhoA活化和肌动蛋白细胞骨架重塑，细胞迁移/侵袭和成肌纤维细胞分化。PRP中的TGF-β 1浓度显著高于PPP。结论PRP和PPP均通过刺激肌动蛋白重塑、激活MMPs、促进细胞迁移和肌成纤维细胞分化，促进创面组织的重塑和收缩。PRP和PPP诱导的相似的生物学反应表明，两种血小板衍生组分可能对牙龈修复产生积极影响。
Objectives Although platelet-rich plasma (PRP) has been proposed as a therapeutic tool to enhance wound repair, the cellular and molecular mechanisms stimulated by this agent are still not completely understood. The present study was designed to characterize the effects of PRP and platelet-poor plasma (PPP) supernatants on cell responses involved in gingival tissue repair. Methods We studied the response of human gingival fibroblasts (HGF) to PRP and PPP fractions on: matrix contraction, cell migration, myofibroblastic differentiation, production of matrix components and proteolytic enzymes. PRP and PPP were obtained from donors using a commercial kit. Matrix contraction was evaluated by means of collagen lattices in the presence of matrix metalloproteinase (MMP) and actin polymerization inhibitors. The production of matrix molecules and proteinases was assessed through Western-blot. RhoA activity was evaluated through a pull-down assay. Actin distribution and focal adhesions were assessed through immunofluorescence. Transforming growth factor-beta (TGF-beta) was quantified through ELISA. Results Both PRP and PPP stimulated human gingival fibroblasts-populated collagen gel contraction and Ilomastat and cytochalasin D inhibited this response. PRP and PPP also stimulated MT1-MMP and TIMP-2 production, RhoA activation and actin cytoskeleton remodeling, cell migration/invasion and myofibroblastic differentiation. TGF-beta 1 was found at significantly higher concentrations in PRP than in PPP. Conclusions Both PRP and PPP promote wound tissue remodeling and contraction through the stimulation of actin remodeling, the activity of MMPs, promotion of cell migration, and myofibroblastic differentiation. The similar biological responses induced by PRP and PPP suggest that both platelet-derived fractions may exert a positive effect on gingival repair.