METHODS: Cre recombinase-induced lineage tracing and cell ablation were conducted in a murine dental implant model. Scratch and transwell assays were used to assess MC3T3-E1 cell migration after paired-related homeobox 1 overexpression. Single-cell RNA sequencing were applied to identify potential genes involved in pairedrelated homeobox 1-positive cells-driven osteogenesis.
RESULTS: Paired-related homeobox 1- positive cells were observed to accumulate in the peri-implant area in a time-dependent manner. The number of these cells were found to reach its maximum on day 14. Osseointegration in mice were noticeably impaired after ablation of paired-related homeobox 1-positive cells. Further, it was discovered that paired-related homeobox 1 promotes MC3T3- E1 cell migration, a process which is indispensable for sound healing of peri-implant tissue. Finally, Semaphorin 3C was detected exclusively and abundantly expressed by paired-related homeobox 1-positive cells. Knockdown of semaphorin 3C in paired-related homeobox 1- positive cells significantly weakened their osteogenic potential.
CONCLUSIONS: Our data suggest that paired-related homeobox 1-positive cells contribute to the osseointegration process under stress stimulation and semaphorin 3C may play a critical role in paired-related homeobox 1- positive cell-driven osteogenesis. Paired-related homeobox 1 could significantly promote MC3T3-E1 cell migration.
方法:Cre重组酶诱导的谱系追踪和细胞消融在鼠牙种植体模型中进行。使用划痕和transwell测定来评估配对相关同源异型盒1过表达后的MC3T3-E1细胞迁移。单细胞RNA测序用于鉴定参与相关同源异型盒1阳性细胞驱动成骨的潜在基因。
结果:观察到配对相关的同源异型盒1-阳性细胞以时间依赖性方式在植入物周围区域积聚。发现这些细胞的数量在第14天达到最大值。切除配对相关的同源盒1阳性细胞后,小鼠的骨整合明显受损。Further,发现配对相关的同源异型盒1促进MC3T3-E1细胞迁移,对于种植体周围组织的声音愈合是必不可少的过程。最后,通过配对相关的同源异型框1阳性细胞专门检测并大量表达了信号蛋白3C。配对相关同源异型盒1-阳性细胞中信号素3C的敲低显着削弱了其成骨潜能。
结论:我们的数据表明,配对相关的同源盒1阳性细胞在应激刺激下促进骨整合过程,而信号素3C可能在配对相关的同源盒1阳性细胞驱动的骨形成中起关键作用。配对相关同源异型盒1能显著促进MC3T3-E1细胞的迁移。