PDF(Pubmed)
Abstract：
The aim of this study is to validate a minimally invasive surgical procedure to harvest palate periosteum as a source of tissue for mesenchymal stromal/stem cells. We performed a standardized procedure to harvest the palate periosteum in ten subjects, which consisted of a 3 mm disposable punch and a Molt periosteal elevator to harvest a small full-thickness fragment of soft tissue at the hard palate area, between the upper bicuspids, 3 to 4 mm apical to the cement enamel junction. The one-third inner portion was fragmented, and following standard cell culture procedures, the adherent cells were cultured for three passages, after obtaining 70-90% confluence. Cell morphology analysis, flow cytometry analysis, and viability and osteogenic differentiation assays were performed. In all 10 cases, uneventful healing was observed, with no need for analgesic intake. The evaluation of cell morphology showed elongated spindle-shaped cells distributed in woven patterns. A high viability range was verified as well as an immunophenotype compatible with mesenchymal stem cell lineage. The differentiation assay showed the potential of the cells to differentiate into the osteogenic lineage. These results demonstrate that the minimally invasive proposed surgical technique is capable of supplying enough periosteum source tissue for stem cell culture and bone tissue engineering.
摘要：
这项研究的目的是验证一种微创手术程序，以收获腭骨膜作为间充质基质/干细胞的组织来源。我们对10名受试者进行了标准化的手术来收获腭骨膜，它由一个3毫米的一次性冲头和一个Molt骨膜升降机组成，用于在硬腭区域收获一个小的全厚度软组织碎片，在上双尖牙之间，水泥牙釉质交界处顶端3～4mm。三分之一的内部是零散的，并遵循标准的细胞培养程序，将贴壁细胞培养三代，在获得70-90%汇合后。细胞形态分析,流式细胞术分析,并进行了活力和成骨分化试验。在所有10个案例中，观察到平静的愈合，不需要服用镇痛药。细胞形态的评估显示细长的纺锤形细胞以编织图案分布。验证了高活力范围以及与间充质干细胞谱系相容的免疫表型。分化测定显示细胞分化为成骨谱系的潜力。这些结果表明，提出的微创手术技术能够为干细胞培养和骨组织工程提供足够的骨膜来源组织。
