Palioto Daniela B, Coletta Ricardo D, Graner Edgard, Joly Julio Cesar, de Lima Antonio Fernando Martorelli
Department of Maxillo-Facial Surgery and Periodontics, School of Dentistry of Ribeirão Preto, University of São Paulo, Brazil.
J Periodontol. 2004 Apr;75(4):498-504. doi: 10.1902/jop.2004.75.4.498.
Enamel matrix derivative (EMD) has recently been shown to promote periodontal regeneration in vivo. Insulin-like growth factor-I (IGF-I) is a potent modulator of periodontal regeneration stimulating cell proliferation, differentiation, synthesis of type I collagen, and non-collagenous proteins. However, the biochemical effects of these factors on periodontal ligament (PDL) fibroblasts are not completely understood. The objective of the present study was to evaluate the effect of EMD, IGF-I, and the combination of these two factors on the proliferation, adhesion, migration, and expression of type I collagen in PDL fibroblasts.
The proliferation rate was measured by automated cell counting and immunohistochemical expression of proliferating cell nuclear antigen (PCNA). The cell adhesion was analyzed by a colorimetric assay and cell migration was measured in Boyden chambers. Type I collagen expression and production was determined by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively.
The results indicated that the proliferation of PDL fibroblasts was significantly stimulated by EMD and EMD plus IGF-I in a dose- and time-dependent manner. EMD, IGF-I, and the combination of both factors had no effects on cellular migration and adhesion or expression and production of type I collagen.
Our results showed that EMD, IGF-I, and the combination of both factors stimulated PDL fibroblast proliferation, whereas these factors did not affect adhesion, migration, or expression of type I collagen of these cells.
牙釉质基质衍生物（EMD）最近已被证明可在体内促进牙周组织再生。胰岛素样生长因子-I（IGF-I）是牙周组织再生的一种有效调节剂，可刺激细胞增殖、分化、I型胶原蛋白及非胶原蛋白的合成。然而，这些因子对牙周膜（PDL）成纤维细胞的生化作用尚未完全明确。本研究的目的是评估EMD、IGF-I以及这两种因子的组合对PDL成纤维细胞增殖、黏附、迁移及I型胶原蛋白表达的影响。
通过自动细胞计数和增殖细胞核抗原（PCNA）的免疫组化表达来测定增殖率。采用比色法分析细胞黏附情况，并在博伊登小室中测量细胞迁移。分别通过半定量逆转录聚合酶链反应（RT-PCR）和酶联免疫吸附测定（ELISA）来确定I型胶原蛋白的表达和产生。
结果表明，EMD以及EMD加IGF-I以剂量和时间依赖性方式显著刺激了PDL成纤维细胞的增殖。EMD、IGF-I以及这两种因子的组合对细胞迁移、黏附或I型胶原蛋白的表达和产生均无影响。
我们的结果表明，EMD、IGF-I以及这两种因子的组合刺激了PDL成纤维细胞的增殖，而这些因子并未影响这些细胞的黏附、迁移或I型胶原蛋白的表达。
