One-stage treatment with autologous red bone marrow transplantation for femoral comminuted fracture

doi:10.3969/j.issn.2095-4344.2017.25.013

Abstract

Abstract:

BACKGROUND: Whether one-stage bone healing with red bone marrow or mesenchymal stem cell transplantation is needed? Whether closed reduction with intramedullary interlocking nailing combined with autologous red bone marrow transplantation can promote osteogenesis in the one-stage treatment of femoral comminuted fracture still remains unclear.
OBJECTIVE: To explore the effects of closed reduction with intramedullary interlocking nailing and one-stage treatment with autologous red bone marrow transplantation on fracture healing.
METHODS: Twenty healthy New Zealand rabbits were selected and anesthetized, and then the models of femoral comminuted fracture were established at the hinder limbs after throwing a 5 kg of hammer from 30 cm height. All rabbits underwent intramedullary fixation using Kirschner wire (diameter: 2 mm); the right limbs served as control groups, and the left limbs as transplantation group, subjected to the injection of autologous red bone marrow (1 mL) into the fracture region. Then the rabbits were respectively killed at 14, and 28 days after modeling, the thickness of thickest callus was measured on X-ray films, and the number of chondrocytes and relative area of bone trabecula in the fracture region were observed through hematoxylin-eosin staining under light microscope.
RESULTS AND CONCLUSION: (1) The thickness of callus formation in the control group was significantly less than that in the transplantation group at 14 days after modeling (P < 0.01). (2) The number of chondrocytes in the transplantation group was significantly more than that in the control group at 14 days after modeling (P < 0.01), and the fracture healing in the transplantation group was faster that in the control group. (3) Compared with the transplantation group, the relative area bone trabecula in the fracture region in the control group was significantly reduced at 28 days after modeling (P < 0.01). (4) These finding indicate that red bone marrow transplantation promotes fracture healing in the intramedullary fixation for femoral comminuted fracture in rabbits.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Femoral Fractures, Models, Animal, Tissue Engineering

CLC Number:

R318

Pan Lin-lin, Wu Hao, Liu Guang-hui, Zhao Zhen-qun. One-stage treatment with autologous red bone marrow transplantation for femoral comminuted fracture[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(25): 4013-4019.

Figures/Tables 1

2.1 实验动物数量分析纳入健康新西兰兔20只，将兔股骨粉碎性骨折内固定造模成功的右侧股骨定义为骨折内固定组，将其左股骨定义为红骨髓移植组。全部进入结果分析，无脱失。 2.2 骨折周围骨痂比较兔股骨骨折髓内钉内固定造模后，对2组兔骨折后及骨折2周时进行X射线检查(图1)，即应用游标卡尺(精确度0.02 mm)测量X射线片上骨折周围骨痂最厚处骨痂厚度，并以影像检查标尺为换算标准，进行计算对应最宽处骨痂厚度，结果显示，骨折内固定组于术后14 d较红骨髓移植组骨折周围骨痂生成厚度明显减小(P < 0.01)，差异有显著性意义，即红骨髓移植组骨折愈合速度快于骨折内固定组，见表1。 2.3 苏木精-伊红染色组织学观察 2.3.1 骨折14 d骨折处组织学变化及软骨细胞生成情况在骨折内固定组，骨折14 d的组织标本中，200倍镜下见局部无菌性炎症反应(图2A)，骨髓腔内出血形成血肿(图2B)，可见新生毛细血管，进而形成肉芽组织，部分标本可见纤维组织(图2C)；400倍镜下见纤维组织中见少量幼小软骨细胞生成。骨折内固定组骨折14 d皆属于处于血肿激化演进期。在红骨髓移植组，骨折14 d的组织标本中，200倍镜下继续可见纤维组织，纤维组织中较多软骨细胞形成(图2D，E)。红骨髓移植组骨折14 d组正处于血肿激化演进期向原始骨痂形成期发展阶段。光镜观察2组骨折14 d骨折处组织切片中软骨细胞数量，计算对应时间点2组骨折处组织切片的软骨细胞浓度，依据前述Pareek的计数方法进行[6]，即每张切片的软骨细胞浓度=5个高倍镜视野(×400)下的软骨细胞数目总和/5。结果显示，骨折内固定组于术后14 d较红骨髓移植组骨折处软骨细胞生成浓度明显稀少(P < 0.01)，差异有显著性意义，即红骨髓移植组骨折处软骨细胞生成数量大于骨折内固定组，骨折愈合速度快于骨折内固定组(见表1)。 2.3.2 骨折28 d骨折处组织学变化及骨小梁生成情况在骨折内固定组，骨折28 d的组织标本中，400倍镜下见骨内外膜可见成骨细胞(图3A)，骨髓腔内的纤维组织及软骨细胞(图3B)，可见纤维组织及软骨组织演进(图3C)。骨折内固定组28 d组皆属于处于原始骨痂形成期初期。在红骨髓移植组，骨折28 d组织的标本中，400倍镜下可见不同成熟程度的骨小梁排列成网梁状(图3D)，与周围纤维软组织相连(图3E)，软骨细胞增生钙化而骨化——软骨内化骨(图3F)。随着软骨细胞钙化成骨，钙化点连接成网状，形成骨小梁，通过骨组织形态计量学分析，镜下观察组织切片，每例均检测新生骨的3个截面，取平均值用于数据分析。测定骨小梁相对面积，指骨小梁(包括矿化骨及类骨质)占全部松质骨(包括骨小梁及髓腔)的面积百分比，属骨量静态参数。结果显示，术后28 d骨折内固定组较红骨髓移植组骨折处骨小梁相对面积明显缩小(P < 0.01)，差异有显著性意义，即红骨髓移植组骨折处骨小梁生成较多，相对面积大于骨折内固定组，骨折愈合速度快于骨折内固定组，见表2。"

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