Human periodontal ligament stem cells (hPDLSCs) sheets play an important role in periodontal tissue engineering. Low‐intensity pulsed ultrasound (LIPUS) has been reported as an effective stimulus to regulate cell biological behavior. The present study aims to explore the potential of LIPUS to promote the formation and function of hPDLSC sheets (hPDLSCSs). Hematoxylin–eosin (H&E) staining, western blot, real‐time PCR, alkaline phosphatase (ALP), and alizarin red staining were used to evaluate the formation and osteogenic effect of LIPUS on hPDLSCSs in vitro. Hydroxyapatite with or without hPDLSCSs was transplanted in the subcutaneous pockets on the back of nude mice and histological analysis was performed. H&E staining showed increased synthesis of extracellular matrix (ECM) and real‐time PCR detected a significant increase in ECM‐related genes after LIPUS treatment. In addition, LIPUS could promote the expression of osteogenic differentiation‐related genes and proteins. ALP and alizarin red staining also found LIPUS enhanced the osteogenesis of hPDLSCSs. After transplantation in vivo, more dense collagen fibers similar to periodontal ligament were regenerated. Collectively, these results indicate that LIPUS not only promotes the formation and osteogenic differentiation of hPDLSCSs but also is a potential treatment strategy for periodontal tissue engineering.

中文翻译：

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低强度脉冲超声促进牙周膜干细胞片的形成和异位牙周组织再生

人牙周膜干细胞 (hPDLSCs) 薄片在牙周组织工程中发挥着重要作用。据报道，低强度脉冲超声（LIPUS）是调节细胞生物学行为的有效刺激物。本研究旨在探索 LIPUS 促进 hPDLSC 片 (hPDLSCS) 形成和功能的潜力。苏木精-伊红 (H&E) 染色、蛋白质印迹、实时荧光定量 PCR、碱性磷酸酶 (ALP) 和茜素红染色用于评估 LIPUS在体外对 hPDLSCSs 的形成和成骨作用. 将带有或不带有 hPDLSCS 的羟基磷灰石移植到裸鼠背部的皮下袋中，并进行组织学分析。H&E 染色显示细胞外基质 (ECM) 合成增加，实时 PCR 检测到 LIPUS 处理后 ECM 相关基因显着增加。此外，LIPUS可以促进成骨分化相关基因和蛋白的表达。ALP 和茜素红染色还发现 LIPUS 增强了 hPDLSCS 的成骨。体内移植后，再生出更致密的类似于牙周韧带的胶原纤维。总之，这些结果表明 LIPUS 不仅促进 hPDLSCSs 的形成和成骨分化，而且是牙周组织工程的潜在治疗策略。