摘要
目的探讨富集骨髓干细胞(bonemarrowstemcells,BMSCs)移植技术促进骨修复的疗效及其影响因素。方法应用富集干细胞技术,在术中完成骨髓血采集、BMSCs富集,通过局部注射或与多孔!-磷酸三钙材料复合后回植,治疗骨缺损患者49例。对富集前后骨髓有核细胞(nucleatedcells,NCs)和碱性磷酸酶染色阳性的细胞集落(colony-formingunitswithalkalinephosphatase+,CFUs/ALP+)计数,分析年龄、性别、病种对骨髓干细胞的影响;通过扫描电镜观察多孔材料与细胞的复合情况;结合影像学检查综合评定疗效。结果平均抽取骨髓血(250±37)ml,经富集技术平均回收(50±19)ml,骨髓成骨源性干细胞回收率78%±18%。其中CFUs/ALP+的自然对数与年龄呈线性负相关(r=-0.4910,P=0.008),NCs与年龄无相关性(r=-0.0469,P=0.8126)。骨髓NCs和CFUs/ALP+的数量无性别差异(t=0.6242,P=0.5397;χ2=2.6512,P=0.1035)。骨折组患者的骨髓CFUs/ALP+数量比退变组多(t=2.2793,P=0.0327),而NCs数量的差异无统计学意义(t=0.7600,P=0.4553)。通过富集,NCs与CFUs/ALP+的增长倍数呈线性正相关(r=0.4054,P=0.0324)。复合2h后,富集BMSCs可在多孔β-磷酸三钙孔内壁贴附。全部病例随访(15.4±6.4)个月,总有效率91.8%。富集血样细菌学检测阴性率100%。术后伤口渗出或肿胀4例,均自行愈合。结论自体富集BMSCs技术是一种快速、简便、安全的临床细胞治疗手段,可与手术同步进行,促进骨修复和再生。年龄、病种因素对BMSCs数量的影响较大。
Objective To explore the clinical effects and influence factors of enriched bone marrow stem cells （BMSCs） graft technique in promoting bone repair. Methods From bone marrow harvested perioperatively, enriched BMSCs obtained by enrichment technique were injected directly or hybrided with β- TCP scaffolds for grafting. 49 patients underwent the treatment. The number of nucleated cells （NCs） was counted and the number of osteoprogenitor cells was estimated by counting the colony-forming units which express alkaline phosphatase （CFUs/ALP＋）. With the analysis of age, gender, disease-related changes in the prevalence of NCs and osteoprogenitor cells of human bone marrow, and the changes of cell number between pre- and post-enrichment, the influence and efficiency of this technique were evaluated. In addition, the adhesion of BMSCs inside scaffolds was observed by SEM. Clinical outcomes were also appraised. Results （250±37） ml bone marrow per patient from bilateral iliac crests was aspirated peri-operatively. About （50±19） ml enriched bone marrow was collected, and 78%±18% of osteoprogenitor cells were recovered. The natural log of CFUs/ALP＋ number declined significantly with age, but not for the Ln （NCs）. The number of NCs and CFUs/ALP＋ was no significantly different between men and women. However the patients with fracture had significantly more CFUs/ALP＋ than those with degenerative disease （t=2.2793, P=0.0327）, but not for the number of NCs （t=0.7600, P=-0.4553）. By enrichment technique, the increasing folds of NCs and CFUs/ALP＋ had a positive line regression （r=0.4054, P=0.0324）. On the other hand, it was identified that enriched BMSCs adhered to the wall of inside porous β-TCP with 2 hours by SEM. The follow-up was about 15.4±6.4 months, 91.8% cases got good results. None of bacteriological examination before grafting was positive. Conclusion The enrichment technique of autologous bone marrow stem cells peri-operatively provide a rapid, simple and safe method for clinical cytotherapy in orthopaedics, which can promote bone repair and regeneration. Age and disease-related changes had perhaps more effects on the number of the BMSCs.
出处
《中华骨科杂志》
CAS
CSCD
北大核心
2006年第11期721-727,共7页
Chinese Journal of Orthopaedics
基金
上海市科委国际合作重点课题(045407008)
关键词
骨髓
干细胞移植
骨再生
Bone marrow
Stem cell transplantation
Bone regeneration
