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. 2017 Feb 2:7:39431.
doi: 10.1038/srep39431.

Repair of bone defects with prefabricated vascularized bone grafts and double-labeled bone marrow-derived mesenchymal stem cells in a rat model

Xiao-Rui Jiang  1 , Hui-Ying Yang  2 , Xin-Xin Zhang  3 , Guo-Dong Lin  1 , Yong-Chun Meng  4 , Pei-Xun Zhang  5 , Shan Jiang  6 , Chun-Lei Zhang  7 , Fei Huang  7 , Lin Xu  4
Affiliations

Repair of bone defects with prefabricated vascularized bone grafts and double-labeled bone marrow-derived mesenchymal stem cells in a rat model

Xiao-Rui Jiang et al. Sci Rep. .

Erratum in

Abstract

This study aims to investigate the repair of bone defects with prefabricated vascularized bone grafts and double-labeled bone marrow-derived mesenchymal stem cells (BMSCs) in a rat model. BMSCs were separated from rat bone marrow. LTR-CMVpro-RFP and LTR-CMVpro-GFP were transfected into the BMSCs for in vitro and in vivo tracking. BMSCs-RFP and BMSCs-GFP were induced into endothelial progenitor cells (EPCs) and osteoblasts (OBs). Rats were divided into five groups: Group A: in vitro prefabrication with EPCs-RFP + in vivo prefabrication with arteriovenous vascular bundle + secondary OBs-GFP implantation; Group B: in vitro prefabrication with EPCs-RFP + secondary OBs-GFP implantation; Group C: in vivo prefabrication with arteriovenous vascular bundle + secondary OBs-GFP implantation; Group D: implantation of EPCs-RFP + implantation of with arteriovenous vascular bundle + simultaneous OBs-GFP implantation; Group E: demineralized bone matrix (DBM) grafts (blank control). Among five groups, Group A had the fastest bone regeneration and repair, and the regenerated bone highly resembled normal bone tissues; Group D also had fast bone repair, but the repair was slightly slower than Group A. Therefore, in vitro prefabrication with EPCs-RFP plus in vivo prefabrication with arteriovenous vascular bundle and secondary OBs-GFP implantation could be the best treatment for bone defect.

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Conflict of interest statement

The authors declare no competing financial interests.

Figures

Figure 1
Figure 1. Positive expressions of VEGFR-2 and CD133.
Note: VEFGR-2, vascular endothelial growth factor receptor 2.
Figure 2
Figure 2. Immunohistochemistry (IHC) of EPCs.
Note: (A) positive vWF expression at the 6th day of incubation; (B) positive vWF expression at the 10th day; (C) positive vWF expression at in the negative control; EPCs, endothelial progenitor cells; vWF, von Willebrand factor.
Figure 3
Figure 3. Morphology of osteoblasts induced with BM-BMSCs (×100).
Note: (A) morphology of osteoblasts induced with BM-BMSCs at the early stage of induction; (B) morphology of osteoblasts induced with BM-BMSCs at the 7th day of induction; (C) morphology of osteoblasts induced with BM-BMSCs at the 12–14th days of induction; BM-BMSCs, bone marrow-derived mesenchymal stem cells.
Figure 4
Figure 4. Identification of induced osteoblasts using immunohistochemical staining.
Note: (A) immunohistochemical staining with bone calcium elements; (B) immunohistochemical staining with Alizanrin Red-s; (C) immunohistochemical staining with tetracycline.
Figure 5
Figure 5. In vitro co-culture of EPCs-RFP and DBM grafts among five groups.
Note: (A) in vitro co-culture of EPCs-RFP and DBM grafts at the 3rd day (×2000); (B) in vitro co-culture of EPCs-RFP and DBM grafts at the 7th day (×1500); (C) in vitro co-culture of EPCs-RFP and DBM grafts at the 10th day (×1000); (D) in vitro co-culture of EPCs-RFP and DBM grafts at the 14th day (×1200).
Figure 6
Figure 6. Histological observation of osteogenesis among the five groups at the 4th, 8th and 12th weeks.
Note: (A) in vitro prefabrication with EPCs-RFP + in vivo prefabrication with arteriovenous vascular bundle + secondary osteoblast (OB)-GFP implantation; (B) in vitro prefabrication with EPCs-RFP + secondary OBs-GFP implantation; (C) in vivo prefabrication with arteriovenous vascular bundle + secondary OBs-GFP implantation; (D) implantation of EPCs-RFP + implantation of with arteriovenous vascular bundle + simultaneous OBs-GFP implantation; (E) only DBM grafts (blank control); ECs, endothelial cells; OB, osteoblast; RFP, red fluorescence protein; GFP, green fluorescence protein.
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