Faculty of Dentistry, University of Toronto, 124 Edward Street, Room 410, Toronto, ON, Canada.
Faculty of Dentistry, University of Toronto, 124 Edward Street, Room 410, Toronto, ON, Canada.
Acta Biomater. 2022 Jul 15;147:209-220. doi: 10.1016/j.actbio.2022.05.035. Epub 2022 May 25.
Common periodontal disease treatment procedures often fail to restore the structural integrity of the junctional epithelium (JE), the epithelial attachment of the gum to the tooth, leaving the tooth-gum interface prone to bacterial colonization. To address this issue, we introduced a novel bio-inspired protein complex comprised of a proline-rich enamel protein, SCPPPQ1, and laminin 332 (LAM332) to enhance the JE attachment. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we showed that SCPPPQ1 and LAM332 interacted and assembled into a protein complex with high-affinity adsorption of 5.9e [M] for hydroxyapatite (HA), the main component of the mineralized tooth surfaces. We then designed a unique shear device to study the adhesion strength of the oral epithelial cells to HA. The SCPPPQ1/LAM332 complex resulted in a twofold enhancement in adhesion strength of the cells to HA compared to LAM332 (from 31 dyn/cm to 63 dyn/cm). In addition, using a modified wound-healing assay, we showed that gingival epithelial cells demonstrated a significantly high migration rate of 2.7 ± 0.24 µm/min over SCPPPQ1/LAM332-coated surfaces. Our collective data show that this protein complex has the potential to be further developed in designing a bioadhesive to enhance the JE attachment and protect the underlying connective tissue from bacterial invasion. However, its efficacy for wound healing requires further testing in vivo. STATEMENT OF SIGNIFICANCE: This work is the first functional study towards understanding the combined role of the enamel protein SCPPPQ1 and laminin 332 (LAM332) in the epithelial attachment of the gum, the junctional epithelium (JE), to the tooth hydroxyapatite surfaces. Such studies are essential for developing therapeutic approaches to restore the integrity of the JE in the destructive form of gum infection. We have developed a model system that provided the first evidence of the strong interaction between SCPPPQ1 and LAM332 on hydroxyapatite surfaces that favored protein adsorption and subsequently oral epithelial cell attachment and migration. Our collective data strongly suggested using the SCPPPQ1/LAM332 complex to accelerate the reestablishment of the JE after surgical gum removal to facilitate gum regeneration.
常见的牙周病治疗程序通常无法恢复牙龈（JE）的结构完整性，JE 是牙龈与牙齿的上皮附着物，使牙齿-牙龈界面容易受到细菌定植。为了解决这个问题，我们引入了一种新型的仿生蛋白复合物，由富含脯氨酸的釉原蛋白 SCPPPQ1 和层粘连蛋白 332（LAM332）组成，以增强 JE 附着。使用带有耗散监测的石英晶体微天平（QCM-D），我们表明 SCPPPQ1 和 LAM332 相互作用并组装成一个具有高亲和力吸附羟基磷灰石（HA）的蛋白复合物，HA 是矿化牙齿表面的主要成分。然后，我们设计了一种独特的剪切装置来研究口腔上皮细胞对 HA 的粘附强度。与 LAM332 相比，SCPPPQ1/LAM332 复合物使细胞对 HA 的粘附强度提高了两倍（从 31 dyn/cm 提高到 63 dyn/cm）。此外，通过改良的划痕愈合试验，我们表明牙龈上皮细胞在 SCPPPQ1/LAM332 涂层表面上表现出显著的高迁移率，为 2.7±0.24 µm/min。我们的综合数据表明，该蛋白复合物具有进一步开发生物粘附物的潜力，以增强 JE 附着并保护下方的结缔组织免受细菌入侵。然而，其在伤口愈合方面的功效需要进一步的体内测试。 意义声明：这项工作是第一项针对理解釉原蛋白 SCPPPQ1 和层粘连蛋白 332（LAM332）在牙龈（JE）与牙齿羟基磷灰石表面的上皮附着中的联合作用的功能研究。这些研究对于开发治疗方法以恢复破坏性牙龈感染中 JE 的完整性至关重要。我们已经开发了一个模型系统，该系统首次提供了 SCPPPQ1 和 LAM332 在羟基磷灰石表面上强烈相互作用的证据，这有利于蛋白质吸附，随后是口腔上皮细胞附着和迁移。我们的综合数据强烈表明，在手术去除牙龈后，使用 SCPPPQ1/LAM332 复合物加速 JE 的重建，以促进牙龈再生。
