Ohnishi Fumio, Ochiai Hiroko, Kishi Kazuo
Department of Plastic and Reconstructive Surgery, Saitama Medical Center, Saitama Medical University, Kawagoe City, Saitama, Japan; Department of Plastic and Reconstructive Surgery, National Hospital Organization Tokyo Medical Center, Meguro-ward, Tokyo, Japan; and Department of Plastic and Reconstructive Surgery, Keio University School of Medicine, Shinjuku-ward, Tokyo, Japan.
Plast Reconstr Surg Glob Open. 2014 Aug 7;2(7):e184. doi: 10.1097/GOX.0000000000000098. eCollection 2014 Jul.
Bone marrow aspirates contain primarily red blood cells. To achieve efficient cell transplantation for regeneration, the red blood cells need to be removed from the aspirates. Cell isolation is typically performed using density gradient centrifugation. However, this method entails issues of clinical safety and convenience. This study describes an efficient method to concentrate bone marrow nucleated cells by hypo-osmotic hemolysis.
The optimal hemolysis conditions were determined by diluting the bone marrow suspensions with distilled water in various dilution ratios. Then, the resulting cell fractions were transplanted in a rat cranial defect model to evaluate their effects on bone formation and their angiogenic effects.
The optimal hemolysis conditions were a 3.3-fold dilution in distilled water and a hypo-osmotic exposure time of 45 seconds. Nucleated cells obtained using this method included granulocytes and mononuclear cells. These cells contain cytoplasmic angiogenic factors, including vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor. In a rat cranial defect model, callus formation and angiogenesis were significantly increased following transplantation of concentrated marrow nucleated cells in this manner.
These results suggest angiogenic and osteogenetic effects of transplanting marrow nucleated cells using this hypo-osmotic method.
骨髓抽吸物主要包含红细胞。为实现用于再生的高效细胞移植，需要从抽吸物中去除红细胞。细胞分离通常采用密度梯度离心法。然而，该方法存在临床安全性和便利性方面的问题。本研究描述了一种通过低渗溶血浓缩骨髓有核细胞的有效方法。
通过用蒸馏水以不同稀释比例稀释骨髓悬液来确定最佳溶血条件。然后，将所得细胞组分移植到大鼠颅骨缺损模型中，以评估它们对骨形成的影响及其血管生成作用。
最佳溶血条件为用蒸馏水3.3倍稀释且低渗暴露时间为45秒。使用该方法获得的有核细胞包括粒细胞和单核细胞。这些细胞含有细胞质血管生成因子，包括血管内皮生长因子、碱性成纤维细胞生长因子和肝细胞生长因子。在大鼠颅骨缺损模型中，以这种方式移植浓缩的骨髓有核细胞后，骨痂形成和血管生成显著增加。
这些结果表明使用这种低渗方法移植骨髓有核细胞具有血管生成和成骨作用。
