The present study aimed to assess the molecular profiles of subepithelial connective tissue grafts (CTGs) obtained at different locations and depths in the human palate. Sixty-four CTGs belonging to anterior deep (AD), anterior superficial (AS), posterior deep (PD), and posterior superficial (PS) groups were subjected to RNA-Sequencing and their transcriptomes were analyzed computationally. Functional correlations characterizing the CTG groups were validated by cell biological experiments using primary human palatal fibroblasts (HPFs) extracted from the CTGs. A clearly more pronounced location-dependent than depth-dependent difference between the grafts, with a minimal number of genes (4) showing no dependence on the location, was revealed. Epithelial, endothelial, and monocytic cell migration was strongly (P < 0.001) potentiated by AD- and PS-HPFs. Moreover, significantly increased expression of genes encoding C-C and C-X-C motif chemokine ligands as well as significantly (P < 0.01) activated p38 signaling suggested immunomodulatory phenotype for AD- and PS-HPFs. Increased growth factor gene expression and significantly activated (P < 0.001) Erk and Akt signaling in HPFs originating from A-CTGs implied their involvement in cell survival, proliferation, and motility. Prominent collagen-rich expression profile contributing to high mechanical stability, increased osteogenesis-related gene expression, and strongly activated (P < 0.001) Smad1/5/8 signaling characterized HPFs originating from P-CTGs. The present data indicate that in humans, differences between palatal CTGs harvested from different locations and depths appear to be location- rather than depth-dependent. Our findings provide the basis for future personalization of the therapeutic strategy by selecting an optimal graft type depending on the clinical indications.

本研究旨在评估在人类上颚不同位置和深度获得的上皮下结缔组织移植物（CTG）的分子特征。对属于前深部 (AD)、前浅部 (AS)、后深部 (PD) 和后浅部 (PS) 组的 64 个 CTG 进行了 RNA 测序，并对它们的转录组进行了计算分析。使用从 CTG 中提取的原代人腭成纤维细胞 (HPF)，通过细胞生物学实验验证了 CTG 组的功能相关性。移植物之间的位置依赖性差异明显比深度依赖性差异更显着，最小数量的基因（4）显示不依赖于位置。AD-和PS-HPF强烈增强上皮细胞、内皮细胞和单核细胞迁移（P  < 0.001）。此外，编码 CC 和 CXC 基序趋化因子配体的基因表达显着增加，并且 p38 信号显着激活（P  < 0.01），表明 AD-和 PS-HPF 具有免疫调节表型。源自 A-CTG 的 HPF 中生长因子基因表达增加并显着激活 ( P  < 0.001) Erk 和 Akt 信号传导表明它们参与细胞存活、增殖和运动。突出的富含胶原蛋白的表达谱有助于高机械稳定性、增加的成骨相关基因表达和强烈激活的 ( P  < 0.001) Smad1/5/8 信号传导是源自 P-CTG 的 HPF 的特征。目前的数据表明，在人类中，从不同位置和深度采集的腭 CTG 之间的差异似乎与位置有关，而不是与深度有关。我们的研究结果为未来个性化治疗策略提供了基础，根据临床适应症选择最佳的移植物类型。