摘要
目的 探索人自体骨髓基质干细胞 (hBMSC)为种子细胞的组织工程骨在治疗临床颅颌面骨缺损中的可行性。方法 自 1999到 2 0 0 2年间 ,选择颅颌面骨缺损病 11例 (外伤性颅骨缺损 4例 ,先天性梨状孔周围骨凹陷畸形 7例 )进行治疗研究。从患者髂前上棘穿刺取骨髓 ,密度梯度离心法分离hBMSC ,经体外成骨诱导和扩增至第 3代。将诱导的hBMSC ,与部分脱钙骨 (partlydemineralizedbonematrix,pDBM)复合 ,并于体外培养一周后 ,手术回植骨缺损区。选择 3例梨状孔凹陷畸形患者 ,在凹陷明显侧植入hBMSC/pDBM复合物 ,对侧轻度凹陷区仅植入单纯pDBM。分别于术后 1,3,6 ,12 ,2 4 ,4 8,5 0个月进行临床外形和三维CT检查随访。 2例患者在Ⅱ期手术时 ,取少量植入物活检 ,行组织学 (HE染色 ) ,免疫组织化学 (骨桥蛋白、骨粘连蛋白 )检测。结果 患者三维CT检查结果示术后 3～ 6月能形成组织工程化骨 ,并修复骨组织缺损。术后 1～ 2 .5年的随访表明组织工程骨稳定存在 ,无明显骨吸收现象 ,临床治疗效果稳定。组织工程骨活检标本HE染色显示其组织学结构与正常松质骨相同 ,并有典型软骨内化骨现象。免疫组织化学显示有骨桥蛋白、骨粘连蛋白阳性表达。而植入的单纯部分脱钙骨于术后 3～ 6月吸收 ,组织学显示为脱钙骨降解碎片和?
Objective To explore the feasibility of tissue engineered bone formation in human being using human bone marrow stromal cells (hBMSCs) and the possibility of clinical repair of craniomaxillofacial bone defects with tissue engineered bone. Methods Total 11 patients of cranial defects and aperture piriformis bone depression were included in this study. The hBMSCs were isolated by Percoll gradient centrifugation from patient′s bone marrow aspirated from iliac crest. The hBMSCs were cultured in vitro and induced to become osteogenic cells in the DMEM medium containing 10% self-serum, beta-glycerophosphate(10 nmol/L) dexamethasone (10 -8 mol/L), L-2-ascorbic acid(50 μmol/L), and 1, 25(OH) 2VD 3(10 nmol/L). Induced hBMSCs of passage 3 were harvested and seeded onto partly demineralized allogenic bone matrix (pDBM) to form a cell-scaffold construct and in vitro co-culture for 1 week. The defects were repaired with the cell-scaffold construct. In 3 cases of aperture piriformis bone depression, one side was repaired with hBMSC / pDBM, while the other side was repaired by pDBM alone. All cases were followed up for 1, 3, 6 months post-operation as short-term evaluation and 1 to 2.5 years post-operation as long-term evaluation by three-dimensional computerized tomography (3D-CT) and clinical examination. In 2 cases who received secondary surgery, extra engineered bone tissue and control pDBM were harvested at the implantation sites for histological examination and immunohistochemistry. Results 3D-CT demonstrated that engineered bone was formed in 3 to 6 months post-operation. Additionally, formed bone maintained stable up to 1-2 years without absorption. Histologically, engineered bones revealed their structures similar to that of normal bone in HE staining. Interestingly, endochondral ossification was also observed in eng-ineered bone. Immunohistochemistry shows positive staining of osteonectin and osteocalcin in engineered and normal bones. In contrast, implanted pDBM was completely degraded in 3-6months as revealed by 3D-CT. Histologically, degraded pDBM and fibrous tissue were observed in the sites where pDBM alone was implanted. Conclusions Tissue engineered bone can be formed in human being. Engineered bone can be used to repair clinical bone defect with satisfactory result. Furthermore, the result of this study proves that tissue engineered bone is possible for clinical application.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2003年第19期1676-1681,共6页
National Medical Journal of China
基金
国家重点基础研究发展规划资助项目 (G19990 5 43 0 4)
国家高科技研究发展规划资助项目(2 0 0 2AA2 0 5 0 11)
上海市科委 (0 0DJ14 0 0 1 1)
