Development of periodontal regeneration using of human alveolar bone periosteal cells

我们研究了使用人牙槽骨骨膜细胞（HABPCs）组成的移植物用于牙周再生的可能性。在牙周部分厚度皮瓣手术中获得牙周组织。从同意自愿参与的患者处获得书面知情同意书，并从该机构的伦理委员会获得伦理审查。用含10%胎牛血清的HABPC培养液（α-MEM）制备细胞片.每周接种HABPCs 4次，形成多层细胞片，折叠后培养3天，形成移植物。采用组织学方法分析移植物形态和蛋白表达（苏木精和伊红，碱性磷酸酶（ALP）和von Kossa染色）和免疫组织化学（I型胶原（Col-I）、骨桥蛋白（OPN）、骨钙素（OC）和Ki-67）的研究。移植物能够容易地用镊子使用，尺寸必须为约5 X 10 mm，并且必须是大约1.5毫米厚。组织学观察表明，移植物仅由细胞和细胞外基质组成。免疫组化染色显示移植物切片呈Col-I、ALP强阳性反应，并可见OC、OPN和Ki-67阳性细胞，提示移植物可能钙化。因此，人们认为，移植物可以应用于牙周再生的骨内缺损。

We studied the possibility of using grafts composed of human alveolar bone periosteal cells (HABPCs) for periodontal regeneration. The periodontium was obtained during periodontal partial thickness flap surgery. Written informed consent was obtained from those who agreed to participate voluntarily and ethical clearance was obtained from the institution's ethical committee. We produced cell sheets using HABPCs with containing 10 % fetal calf serum medium (α-MEM). HABPCs was inoculated four times to become the multilayer cell sheet every week that was cultured for three days after folded to create the graft. The graft morphology and protein expression were analyzed using histological (hematoxylin and eosin, alkaline phosphatase (ALP) and von Kossa's staining) and immunohistochemical (type I collagen (Col-I), osteopontin (OPN), osteocalcin (OC) and Ki-67) studies on the graft sections.The graft be able to be used with tweezers easily, must be about 5 X 10 mm in size, and must be about 1.5 mm thickness. As a result of the histological observation, the graft was consisted only of the cell and the extracellular matrix. Immunohistochemical staining of the graft sections showed a strong positive reaction for Col-I, ALP and included OC, OPN and Ki-67 positive cell.It was suggested that graft might be calcified by the above-mentioned results. Therefore, it was thought that the graft could be applied to an intraosseous defect for periodontal regeneration.