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Transplanted Endothelial Cells Enhance Orthotopic Bone Regeneration

¹ Depts. of Periodontics/Prevention/Geriatrics,
² Biologic and Materials Sciences, and
³ Chemical and Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; and
⁴ Div. of Engineering and Applied Sciences, Harvard University, 29 Oxford St., 325 Pierce Hall, Cambridge, MA 02138, USA

View larger version (26K): [in a new window]   Figure 1. Blood vessel analysis. (a) Bone marrow stromal cells were transplanted with (BMSC/EC) and without endothelial cells (BMSC). Six weeks following transplantation, the total numbers of blood vessels were counted and normalized per unit area following retrieval of implants. (b) Human-derived vessels were determined by human CD31(+) staining of vessels and plotted as percentages of total functional vessels. (c) 12 weeks following transplantation, total blood vessels were counted in both conditions, and an additional group, control implant condition of poly-lactide-co-glycolide scaffolds that did not contain any cells (PLGA), was also included at this time-point. (d) Human-derived vessels were determined by CD31(+) stained vessels and expressed as percentages of total vessels. Values represent mean ± standard error of the means (unless otherwise noted). *Statistically significant difference (p < 0.05), as compared with BMSC condition; **statistically significant difference as compared with both of the other experimental conditions (n = 6).

View larger version (52K): [in a new window]   Figure 2. Bone analysis at 6 wks. Photomicrographs of hematoxylin & eosin (H & E)-stained sections retrieved 6 wks after transplantation of (a) bone marrow stromal cells alone (BMSC) or (b) with endothelial cells (BMSC/EC). (c) Following retrieval of the scaffolds used to transplant the cells, total bone volume was quantified at this time-point by µCT. (d) Bone mineral density was also quantified. Values represent mean ± standard error of the means (unless otherwise noted). *Statistically significant difference (p < 0.05), as compared with BMSC condition. Size bars are shown in (a) and (b) (n = 6).

View larger version (46K): [in a new window]   Figure 3. Calvarial defects at 12 wks. (a,c) Photomicrographs (H & E-stained) and (b,d) µCT images of cross-sections of the calvarial defect sites implanted with (a,b) bone marrow stromal cells alone and (c,d) co-transplantated with endothelial cells. Arrows demarcate defect margins (8.5-mm defect). Size bars are shown in (a) and (c).

View larger version (27K): [in a new window]   Figure 4. Bone formation at 12 wks. (a) Photomicrograph of an H & E-stained section from a co-transplant sample retrieved at 12 wks. Micro-CT analyses of (b) bone mineral density from bone marrow stromal cell transplants (BMSC) and bone marrow stromal cell, endothelial cell co-transplants (BMSC/EC). (c) Total bone volume fraction in BMSC transplants and co-transplants of BMSC and EC. An additional experimental group of scaffolds alone with no cells (PLGA) was included at this time-point. A scale bar is shown in (a). Values represent mean ± standard error of the mean. *Statistically significant difference (p < 0.05), as compared with BMSC condition (n = 6).