Original Full Length ArticleTransplantation of hypoxia preconditioned bone marrow mesenchymal stem cells enhances angiogenesis and osteogenesis in rabbit femoral head osteonecrosis
Introduction
Osteonecrosis of the femoral head (ONFH) is a difficult and complicated orthopedic disorder that often leads to femoral head collapse with activity-related hip pain [1]. Several treatment methods for ONFH have been attempted, including drug therapy, surgical core decompression, vascularized bone grafting, and osteotomy. However, these methods have barely been effective in treating the disease. Although the pathogenesis and etiology of ONFH have not yet been completely understood, the interrupted blood circulation in the femoral head that leads to osseous tissue necrosis seems to be the major factor [2], [3], [4]. Determining how to promote vascular repair and angiogenesis in ONFH becomes a crucial part of the treatment.
Bone marrow mesenchymal stem cells (BMMSCs) are multipotent adult stem cells that are able to differentiate into several cell lineages, including endothelial cells and osteocytic cells. Both animal and clinical studies have provided evidence that BMMSC transplantation has potency in the treatment of ischemic diseases, such as myocardial infarction and ONFH [5], [6]. A major limitation in stem cell therapy for ischemic diseases is the low survival rate of transplanted cells in the ischemic and peri-necrosis region [7]. Thus, improving grafted cell survival is vital for enhancing the efficacy and efficiency of stem cell therapy. However, Hernigou and Beaujean demonstrated that abnormal proliferation and differentiation of MSCs led to osteonecrosis of the femoral head [8]. Later, Wei et al. suggested that glucocorticoid has a direct effect on impairing the proliferation and function of BMMSCs [9]. It would be helpful if we discovered a new way to reverse the impairment of osteonecrosis BMMSCs for treating ONFH.
Hypoxic preconditioning (HP) by sublethal hypoxic insult stimulates endogenous mechanisms resulting in compensatory regulation including protein expressions that protect against future lethal hypoxia and other insults [10]. On the basis of the well-documented manifold benefits of HP, in the present investigation, we hypothesize that the HP approach in BMMSC transplantation therapy can lead to a set of multiple responses that increase trophic factor support, reduce cell apoptosis, stimulate endothelial and osteogenic differentiation, and finally reverse the impairment of osteonecrosis BMMSCs. A combination of these effects will result in increased angiogenesis, osteogenesis, and ultimately better bone tissue repair after osteonecrosis.
Section snippets
The rabbit model of early ONFH
Twenty-eight 26–28-week old male mature New Zealand White rabbits with body weight of 3.2–4.0 kg for each group (model group, 20% O2 BMMSC group, and 2% O2 BMMSC group) were used in this experiment. The rabbits were randomly assigned to the groups in our study. All investigators were blinded to the treatment group for all analyses.
The ONFH induction procedure was performed using the following method. First, lipopolysaccharide (LPS; Sigma, USA) (10 μg/kg) was injected through an ear vein.
Characterization of rabbit BMMSCs
BMMSCs were successfully expanded three days after initial seeding, and they rapidly expanded into colonies of confluent spindle cells at ten to fourteen days. The third-passage cells were incubated with antibodies of both CD105 and CD34. They were chosen as markers in flow cytometry. The results showed that the cells were positive to CD105 (99.12%) and negative to CD34 (0.88%). The cultured cells were effectively BMMSCs.
Histological analysis of the ONFH model group and core decompression groups
The model group showed discrete trabecular bone with many empty lacunae,
Discussion
Osteonecrosis of the femoral head (ONFH) is a relatively common disease. Because the pathogenesis and etiology of nontraumatic osteonecrosis of the femoral head (ONFH) has not been completely understood, current treatment of ONFH simply focuses on preventing irreversible complications, such as the biomechanical collapse of the femoral head and osteoarthritis of the hip joint. However, in many cases, these treatments have not been effective in preventing the disease [17].
Previous studies
Conflicts of interest
Lihong Fan, Chen Zhang, Zefeng Yu, Zhibin Shi, Xiaoqian Dang, and Kunzheng Wang declare that they have no conflict of interest.
Acknowledgments
This research was supported by the National Natural Science Foundation of China (Grant nos. 81101363, 81371944, 81301562 and 81572145) and the Fundamental Research Funds for the Central Universities.
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