Warnke Patrick H, Douglas Timothy, Sivananthan Sureshan, Wiltfang Jörg, Springer Ingo, Becker Stephan T
Department of Oral and Maxillofacial Surgery, University of Kiel, Kiel, Germany.
Clin Oral Implants Res. 2009 Aug;20(8):761-6. doi: 10.1111/j.1600-0501.2009.01709.x. Epub 2009 Mar 27.
The cultivation of bone is a major focus in tissue engineering and oral implantology. Without a periosteal layer, instant or rapid development of a substantial cortical layer is unlikely for engineered bone grafts. The aim of this study was to test the ability of four collagen membranes to support and promote the proliferation of human periosteal cells.
Human periosteum cells were cultured using an osteogenic medium consisting of Dulbecco's modified Eagle's medium supplemented with fetal calf serum, penicillin, streptomycin and ascorbic acid at 37 degrees C with 5% CO(2). Four collagen membranes served as scaffolds: Bio-Gide, Chondro-Gide, Tutodent and Ossix Plus. Cell vitality was assessed by fluorescin diacetate (FDA) and propidium iodide (PI) staining, biocompatibility with LDH and BrdU, MTT, WST tests and scanning electron microscopy (SEM).
After 24 h, all probes showed viable periosteal cells. All biocompatibility tests revealed that proliferation on all membranes after treatment with eluate from membranes after a 24-h immersion in a serum-free cell culture medium was similar to the controls. Periosteal cells formed layers covering the surfaces of all four membranes 7 days after seeding in SEM.
It can be concluded from our data that the collagen membranes can be used as scaffolds for the cultivation of periosteum layers with a view to creating cortical bone using tissue-engineering methods.
骨组织培养是组织工程学和口腔种植学的主要研究重点。缺乏骨膜层的情况下，工程化骨移植不太可能立即或快速形成大量皮质层。本研究旨在测试四种胶原膜支持和促进人骨膜细胞增殖的能力。
使用由杜氏改良 Eagle 培养基组成的成骨培养基培养人骨膜细胞，该培养基添加了胎牛血清、青霉素、链霉素和抗坏血酸，在 37℃、5%二氧化碳条件下培养。四种胶原膜用作支架：Bio-Gide、Chondro-Gide、Tutodent 和 Ossix Plus。通过荧光素二乙酸酯（FDA）和碘化丙啶（PI）染色、与乳酸脱氢酶（LDH）和溴脱氧尿苷（BrdU）的生物相容性、MTT、WST 试验以及扫描电子显微镜（SEM）评估细胞活力。
24 小时后，所有探针均显示骨膜细胞存活。所有生物相容性测试表明，在无血清细胞培养基中浸泡 24 小时后，用膜洗脱液处理后，所有膜上的细胞增殖与对照组相似。在扫描电子显微镜下，接种 7 天后，骨膜细胞在所有四种膜的表面形成了层状结构。
从我们的数据可以得出结论，胶原膜可作为培养骨膜层的支架，以期利用组织工程方法构建皮质骨。
