Hospital Sírio-Libanês, Instituto de Ensino e Pesquisa, São Paulo, SP, Brazil.
Hospital Municipal Infantil Menino Jesus, São Paulo, SP, Brazil.
Stem Cell Res Ther. 2020 Nov 25;11(1):501. doi: 10.1186/s13287-020-02017-7.
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 mm), 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.
先天性颅面畸形（影响约 2-3%的新生儿）的骨重建一直是骨组织工程领域的研究重点。间充质基质细胞在再生医学方案中的可能性开辟了一个新的研究领域，旨在寻找最佳的多能细胞来源，可以通过非侵入性程序分离。在这项研究中，我们分析了提上唇肌碎片（可在腭裂患者的腭裂修复手术中通过非侵入性方式轻松获得）是否代表具有成骨潜力的 MSC 的新来源。
我们从 5 名无关患者的腭裂修复手术中获得了提上唇肌碎片（3-5mm）。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。通过流式细胞术分析和沿成骨、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，将它们用于构建免疫功能正常的大鼠全层颅骨缺损模型的临界尺寸。
流式细胞术分析表明，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）或内皮细胞标志物（CD31）呈阴性。在适当的细胞培养条件下，细胞成功地进行了成骨、软骨和成脂细胞分化。用分离的提上唇肌细胞接种 CellCeram™支架的颅骨缺损比用无细胞支架治疗的颅骨缺损显示出更大的骨愈合。
提上唇肌衍生的细胞在体外和体内均具有与其他间充质基质细胞相似的表型特征。我们的研究结果表明，这些细胞在腭裂和其他以显著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。
