Department of Orthopaedic Surgery, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan.
Clin Orthop Relat Res. 2010 Oct;468(10):2725-33. doi: 10.1007/s11999-010-1444-z. Epub 2010 Jun 24.
Cell therapy using autologous cells has been used in the treatment of various medical conditions. The mononuclear cell (MNC) fraction of bone marrow (BM) contains stem/progenitor cells that could contribute to osteogenesis and angiogenesis.
QUESTIONS/PURPOSES: We asked whether MNCs derived from intraoperative shed blood (SB), consisting of peripheral blood and BM, have osteoinductive and angiogenic potential.
We harvested SB and BM from six patients undergoing THA. Isolated MNCs from SB and BM were analyzed by flow cytometry to evaluate the CD34(+) cell fraction and 1 × 10(6) cells were seeded on an interconnective porous calcium hydroxyapatite ceramic (IP-CHA) and transplanted in the backs of athymic rats. IP-CHAs without cells were transplanted as controls and all composites were harvested after 4 and 8 weeks. Osteoinductive potential was evaluated by histologic observation, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA) using anti-osteocalcin (OC) antibodies qualitatively and quantitatively. To evaluate angiogenic potential, capillary density was measured by immunohistochemistry using Isolectin B4 4 weeks after implantation.
We found that CD34(+) cells existed in SB-MNCs and there was a trend toward lower frequency compared with BM-MNCs. Histologic osteoinduction, OC expression, and capillary density were increased by transplantation of MNCs from SB. Similar results were achieved with MNCs from BM.
MNCs from SB have equivalent osteoinductive and angiogenic potential compared with those from BM.
SB could be an attractive source for isolation of MNCs, enhancing osteoinduction and neovascularization, to augment the reconstruction of skeletal defects.
利用自体细胞的细胞疗法已被用于治疗各种医学病症。骨髓(BM)的单核细胞(MNC)部分含有可能有助于成骨和血管生成的干细胞/祖细胞。
问题/目的:我们想知道源自术中失血(SB)的 MNC (由外周血和 BM 组成)是否具有成骨和血管生成的潜力。
我们从六名接受 THA 的患者中采集 SB 和 BM。通过流式细胞术分析从 SB 和 BM 中分离的 MNC,以评估 CD34+细胞分数,并将 1×106 个细胞接种在互连多孔钙羟基磷灰石陶瓷(IP-CHA)上,并移植到无胸腺大鼠的背部。没有细胞的 IP-CHAs 被移植作为对照,所有复合材料在 4 和 8 周后被采集。通过组织学观察、免疫组织化学和酶联免疫吸附试验(ELISA)用抗骨钙素(OC)抗体定性和定量评估成骨潜力。为了评估血管生成潜力,在植入后 4 周通过免疫组织化学用异硫氰酸荧光素 B4 测量毛细血管密度。
我们发现 CD34+细胞存在于 SB-MNC 中,与 BM-MNC 相比,其频率呈下降趋势。来自 SB 的 MNC 移植可增加组织学成骨诱导、OC 表达和毛细血管密度。来自 BM 的 MNC 也取得了类似的结果。
与 BM 相比,来自 SB 的 MNC 具有等效的成骨和血管生成潜力。
SB 可能是分离 MNC 的有吸引力的来源,可增强成骨和新血管生成,以增强骨骼缺陷的重建。