Bone Regeneration Using Autologous Adipose-Derived Stem Cell Spheroid Complex
Bone Regeneration Using Autologous Adipose-Derived Stem Cell Spheroid Complex
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使用自体脂肪干细胞球体复合物进行骨再生
DOI：
10.1166/jbt.2022.3014
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Toshiaki Naka
中科院分区：
文献类型：
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作者：
Katsuhiro Hayashi;Xang Fang;Hiroshi Ueda;Akihiro Miwa;Toshiaki Naka
骨缺损需要使用各种生物材料或非生物材料进行重建。干细胞球体可用于无支架入路成骨。我们建立了一种培养方法，通过测量蛋白质在一系列培养基中的表达来创造最佳的成骨脂肪源性干细胞（ADSC）球体复合物。ADSC球体在DMEM中培养24小时后，在含抗坏血酸的培养基中培养5天，然后在成骨细胞分化培养基中培养。在换入成骨细胞分化培养基1天后，收集球状体，培养4天，获得球状复合体。通过分析I型胶原、碱性磷酸酶和整合素α5的表达来确定每个培养周期，以最大限度地提高ADSC球体的活性。I型胶原蛋白在含抗坏血酸培养基中表达显著增加（p α5表达）。在成骨细胞分化培养的第一天，表达量短暂升高，随后逐渐降低。成骨细胞分化培养基增强球状体细胞黏附。一项体内研究证实了由既定方案产生的ADSC球体复合物的成骨潜力。ADSC球体复合物刺激骨再生，并将应用于大骨缺损的治疗。
Bone defects require reconstruction using various biomaterials or non-biological materials. Stem cell spheroids can be used for scaffold-free approaches for osteogenesis. We set up a culture method for creating an optimal osteogenic adipose-derived stem cell (ADSC) spheroid complex by measuring the expression of protein in a sequential series of culture media. After culturing ADSC spheroids for 24 hours in DMEM, the spheroids were cultured in ascorbic acid-containing medium for five days followed by osteoblast differentiation medium. One day after exchanging to osteoblast differentiation medium, spheroids were collected and cultured for four days to obtain a spheroid complex. Each culture period was determined by analyzing the expression of collagen type I, alkaline phosphatase and integrin α5 to maximize the activity of ADSC spheroids. The expression of collagen type I increased significantly in ascorbic acid-containing medium (p α5 expression. The expression was transiently high on the first day of osteoblast differentiation culture and then gradually decreased. Osteoblast differentiation medium enhanced cell adhesion in spheroids. An in vivo study confirmed the osteogenic potential of the ADSC spheroid complex created by the established protocol. The ADSC spheroid complex stimulated bone regeneration and will be applied to the treatment of large bone defects.