Division of Endocrinology and Centre for Research in Anabolic Skeletal Targets in Health and Illness (ASTHI), CSIR-Central Drug Research Institute, B.S. 10/1, Sector-10, Jankipuram Extension, Lucknow, India.
Stem Cell Rev Rep. 2015 Apr;11(2):309-21. doi: 10.1007/s12015-014-9573-5.
We investigated deleterious changes that take place in mesenchymal stem cells (MSC) and its fracture healing competence in ovariectomy (Ovx)-induced osteopenia. MSC from bone marrow (BM) of ovary intact (control) and Ovx rats was isolated. (99m)Tc-HMPAO (Technitium hexamethylpropylene amine oxime) labeled MSC was systemically transplanted to rats and fracture tropism assessed by SPECT/CT. PKH26 labeled MSC (PKH26-MSC) was bound in scaffold and applied to fracture site (drill-hole in femur metaphysis). Osteoinduction was quantified by calcein binding and microcomputed tomography. Estrogen receptor (ER) antagonist, fulvestrant was used to determine ER dependence of osteo-induction by MSC. BM-MSC number was strikingly reduced and doubling time increased in Ovx rats compared to control. SPECT/CT showed reduced localization of (99m)Tc-HMPAO labeled MSC to the fracture site, 3 h post-transplantation in Ovx rats as compared with controls. Post-transplantation, Ovx MSC labeled with PKH26 (Ovx PKH26-MSC) localized less to fracture site than control PKH26-MSC. Transplantation of either control or Ovx MSC enhanced calcein binding and bone volume at the callus of control rats over placebo group however Ovx MSC had lower efficacy than control MSC. Fulvestrant blocked osteoinduction by control MSC. When scaffold bound MSC was applied to fracture, osteoinduction by Ovx PKH26-MSC was less than control PKH26-MSC. In Ovx rats, control MSC/E2 treatment but not Ovx MSC showed osteoinduction. Regenerated bone was irregularly deposited in Ovx MSC group. In conclusion, Ovx is associated with diminished BM-MSC number and its growth, and Ovx MSC displays impaired engraftment to fracture and osteoinduction besides disordered bone regeneration.
我们研究了去卵巢（Ovx）诱导骨质疏松症对骨髓间充质干细胞（MSC）及其骨折愈合能力的有害变化。从卵巢完整（对照）和 Ovx 大鼠的骨髓（BM）中分离 MSC。将（99m）Tc-HMPAO（Technitium 六甲基丙二胺肟）标记的 MSC 系统移植到大鼠体内，并通过 SPECT/CT 评估骨折趋化性。将 PKH26 标记的 MSC（PKH26-MSC）结合到支架上，并应用于骨折部位（股骨干骺端的钻孔）。通过钙黄绿素结合和微计算机断层扫描来量化成骨诱导。使用雌激素受体（ER）拮抗剂氟维司群来确定 MSC 成骨诱导的 ER 依赖性。与对照组相比，Ovx 大鼠的 BM-MSC 数量明显减少，倍增时间延长。与对照组相比，Ovx 大鼠在移植后 3 小时，（99m）Tc-HMPAO 标记的 MSC 向骨折部位的定位减少。与对照组相比，移植后，Ovx PKH26-MSC 向骨折部位的定位较少。与安慰剂组相比，无论是移植对照 MSC 还是 Ovx MSC 都增强了对照大鼠骨痂的钙黄绿素结合和骨体积，但 Ovx MSC 的疗效低于对照 MSC。氟维司群阻断了对照 MSC 的成骨诱导。当支架结合的 MSC 应用于骨折时，Ovx PKH26-MSC 的成骨诱导作用低于对照 PKH26-MSC。在 Ovx 大鼠中，对照 MSC/E2 治疗而不是 Ovx MSC 显示出成骨诱导作用。在 Ovx MSC 组中，再生的骨不规则沉积。总之，Ovx 与 BM-MSC 数量及其生长减少有关，并且 Ovx MSC 显示出向骨折部位的植入受损和成骨诱导受损，此外还有骨骼再生紊乱。
