Bone reconstruction in congenital craniofacial differences, which affect about 2–3% of newborns, has long been the focus of intensive research in the field of bone tissue engineering. The possibility of using mesenchymal stromal cells in regenerative medicine protocols has opened a new field of investigation aimed at finding optimal sources of multipotent cells that can be isolated via non-invasive procedures. In this study, we analyzed whether levator veli palatini muscle fragments, which can be readily obtained in non-invasive manner during palatoplasty in cleft palate patients, represent a novel source of MSCs with osteogenic potential. We obtained levator veli palatini muscle fragments (3–5 mm3), during surgical repair of cleft palate in 5 unrelated patients. Mesenchymal stromal cells were isolated from the muscle using a pre-plating technique and other standard practices. The multipotent nature of the isolated stromal cells was demonstrated via flow cytometry analysis and by induction along osteogenic, adipogenic, and chondrogenic differentiation pathways. To demonstrate the osteogenic potential of these cells in vivo, they were used to reconstruct a critical-sized full-thickness calvarial defect model in immunocompetent rats. Flow cytometry analysis showed that the isolated stromal cells were positive for mesenchymal stem cell antigens (CD29, CD44, CD73, CD90, and CD105) and negative for hematopoietic (CD34 and CD45) or endothelial cell markers (CD31). The cells successfully underwent osteogenic, chondrogenic, and adipogenic cell differentiation under appropriate cell culture conditions. Calvarial defects treated with CellCeram™ scaffolds seeded with the isolated levator veli palatini muscle cells showed greater bone healing compared to defects treated with acellular scaffolds. Cells derived from levator veli palatini muscle have phenotypic characteristics similar to other mesenchymal stromal cells, both in vitro and in vivo. Our findings suggest that these cells may have clinical relevance in the surgical rehabilitation of patients with cleft palate and other craniofacial anomalies characterized by significant bone deficit.

中文翻译：

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人提肌veli palatini肌肉：间充质基质细胞的一种新来源，用于先天性颅面畸形患者的康复

早在颅骨组织工程领域，深入研究的先天性颅面差异的骨重建就一直困扰着大约2％至3％的新生儿。在再生医学方案中使用间充质基质细胞的可能性开辟了一个新的研究领域，旨在寻找可以通过非侵入性程序分离的多能细胞的最佳来源。在这项研究中，我们分析了c裂患者在plast裂术中可以容易地以无创方式获得的提肌vela palatini肌肉碎片是否代表了具有成骨潜能的MSC的新来源。在5例无关患者的surgical裂手术修复中，我们获得了提肛veli palatini肌肉碎片（3-5 mm3）。使用预铺板技术和其他标准方法从肌肉中分离间充质基质细胞。通过流式细胞仪分析和沿成骨，成脂和成软骨分化途径的诱导证明了分离的基质细胞的多能性。为了证明这些细胞在体内的成骨潜力，将它们用于在免疫活性大鼠中重建临界大小的全厚度颅盖骨缺损模型。流式细胞仪分析显示，分离的基质细胞对间充质干细胞抗原（CD29，CD44，CD73，CD90和CD105）呈阳性，对造血（CD34和CD45）或内皮细胞标记（CD31）呈阴性。在适当的细胞培养条件下，细胞成功地经历了成骨，成软骨和成脂细胞分化。与用脱细胞支架治疗的缺损相比，用分离的提肌veli palatini肌细胞播种的CellCeram™支架处理的颅骨缺损显示出更好的骨愈合。在体外和体内，源自提肛肌的细胞具有与其他间充质基质细胞相似的表型特征。我们的发现表明，这些细胞在in裂和其他以严重骨缺损为特征的颅面畸形患者的手术康复中可能具有临床意义。无论是体内还是体外。我们的发现表明，这些细胞在c裂和其他以严重骨缺损为特征的颅面畸形患者的手术康复中可能具有临床意义。无论是体内还是体外。我们的发现表明，这些细胞在c裂和其他以严重骨缺损为特征的颅面畸形患者的手术康复中可能具有临床意义。