An indispensable role of macrophages in bone repair has been well recognized. Previous data have demonstrated the copresence of M1 macrophages and mesenchymal stem cells (MSCs) during the proinflammatory stage of bone repair. However, the exact role of M1 macrophages in MSC function and bone repair is unknown. This study aimed to define the role of M1 macrophages at bone injury sites via the function of 1,25-Dihydroxyvitamin D (1,25[OH] ₂ D) in suppressing M1 but promoting M2 differentiation. We showed that 1,25(OH) ₂ D suppressed M1 macrophage–mediated enhancement of MSC migration. Additionally, 1,25(OH) ₂ D inhibited M1 macrophage secretion of osteogenic proteins (i.e., Oncostatin M, TNF-α, and IL-6). Importantly, the 1,25(OH) ₂ D-mediated suppression of osteogenic function in M1 macrophages at the proinflammatory stage was associated with 1,25(OH) ₂ D-mediated reduction of MSC abundance, compromised osteogenic potential of MSCs, and impairment of fracture repair. Furthermore, outside the proinflammatory stage, 1,25(OH) ₂ D treatment did not suppress fracture repair. Accordingly, our data support 2 conclusions: (a) M1 macrophages are important for the recruitment and osteogenic priming of MSCs and, hence, are necessary for fracture repair, and (b) under vitamin D–sufficient conditions, 1,25(OH) ₂ D treatment is unnecessary and can be detrimental if provided during the proinflammatory stage of fracture healing.

Reconstruction of critical sized bone injuries is a major problem that continues to inspire the design of new materials and grafts. Natural ceramics (hydroxyapatite (HA) coralline HA, or synthetic HA) and β-tricalcium phosphate (β-TCP) are being explored for use as scaffolds in bone tissue engineering, among several other materials. The present study evaluated the bone forming capacity of nanosize bioceramics synthesized in situ in poly-vinyl alcohol (PVA) with different ratios of HA and β-TCP; the Ca/P ratio was 1.62 for bioceramic P1, 1.60 for P2 and 1.58 for P3. Further osteogenesis in vitro with mesenchymal stem cells (MSC) acquired from different sources for osteogenesis in vitro and their bone healing properties in vivo were also evaluated. MSC isolated from human placenta, Wharton’s jelly from umbilical cord, fetal bone marrow and adipose tissue, cultured in the presence of nanosize bioceramic particles, were monitored for osteogenic differentiation. Placental cells showed the best osteogenic potential of the different MSC studied on the basis of expression of osteogenic markers. Complete regeneration of the damaged region was observed in vivo when MSC derived from placenta were used with nanoceramic (Ca/P ratio 1.58) in the experimental defect created in the femur of Wistar rats. Even small variation in the Ca/P ratio can alter the outcome of tissue constructs.

Lineage-specific expansion of haematopoietic stem/progenitor cells (HSPCs) from human umbilical cord blood (UCB) is desirable because of their several applications in translational medicine, e.g. treatment of cancer, bone marrow failure and immunodeficiencies. The current methods for HSPC expansion use either cellular feeder layers and/or soluble growth factors and selected matrix components coated on different surfaces. The use of cell-free extracellular matrices from bone marrow cells for this purpose has not previously been reported. We have prepared insoluble, cell-free matrices from a murine bone marrow stromal cell line (MS-5) grown under four different conditions, i.e. in presence or absence of osteogenic medium, each incubated under 5% and 20% O₂ tensions. These acellular matrices were used as biological scaffolds for the lineage-specific expansion of magnetically sorted CD34⁺ cells and the results were evaluated by flow cytometry and colony-forming assays. We could get up to 80-fold expansion of some HSPCs on one of the matrices and our results indicated that oxygen tension played a significant role in determining the expansion capacity of the matrices. A comparative proteomic analysis of the matrices indicated differential expression of proteins, such as aldehyde dehydrogenase and gelsolin, which have previously been identified as playing a role in HSPC maintenance and expansion. Our approach may be of value in identifying factors relevant to tissue engineering-based ex vivo HSPC expansion, and it may also provide insights into the constitution of the niche in which these cells reside in the bone marrow. Copyright © 2012 John Wiley & Sons, Ltd.

This study identifies that cyclooxygenase 2 targets CD90 ⁺ skeletal stem cells in fracture sites to accelerate fracture repair.

Current therapies for gut inflammation have not reached the desired specificity and are attended by unintended immune suppression. This study aimed to provide evidence for supporting a hypothesis that direct in vivo augmentation of the induction of gut-homing regulatory T (Treg) cells is a strategy of expected specificity for the treatment of chronic intestinal inflammation (e.g., inflammatory bowel disease). We showed that dendritic cells (DCs), engineered to de novo produce high concentrations of both 1,25-dihydroxyvitamin D, the active vitamin D metabolite, and retinoic acid, an active vitamin A metabolite, augmented the induction of T cells that express both the regulatory molecule Foxp3 and the gut-homing receptor CCR9 in vitro and in vivo. In vivo, the newly generated Ag-specific Foxp3+ T cells homed to intestines. Additionally, transfer of such engineered DCs robustly suppressed ongoing experimental colitis. Moreover, CD4+ T cells from spleens of the mice transferred with the engineered DCs suppressed experimental colitis in syngeneic hosts. The data suggest that the engineered DCs enhance regulatory function in CD4+ T cell population in peripheral lymphoid tissues. Finally, we showed that colitis suppression following in vivo transfer of the engineered DCs was significantly reduced when Foxp3+ Treg cells were depleted. The data indicate that maximal colitis suppression mediated by the engineered DCs requires Treg cells. Collectively, our data support that DCs de novo overproducing both 1,25-dihydroxyvitamin D and retinoic acid are a promising novel therapy for chronic intestinal inflammation.