Long bone fracture repair in mice harboring GFP reporters for cells within the osteoblastic lineage

Ushiku, Chikara; Adams, Douglas; Jiang, Xi; Wang, Liping; Rowe, David W.

doi:10.1002/jor.21105

Cited by 62 publications

(60 citation statements)

References 47 publications

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“…Fixed samples were processed into cryoblocks, and nondecalcified sections (5 lm) were obtained to qualitatively observe repair, the mineral deposition pattern visualized by bone labels, and alkaline phosphatase (AP) staining for osteoblastic activity [21,22,41,44]. Slides were imaged at 9 5 magnification and high-resolution image mosaics were acquired.…”

Section: Vital Bone Labelsmentioning

confidence: 99%

Hydrogel-based Delivery of rhBMP-2 Improves Healing of Large Bone Defects Compared With Autograft

Krishnan

Priddy

Esancy

et al. 2015

Clinical Orthopaedics &Amp; Related Research

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Background Autologous bone grafting remains the gold standard in the treatment of large bone defects but is limited by tissue availability and donor site morbidity. Recombinant human bone morphogenetic protein-2 (rhBMP-2), delivered with a collagen sponge, is clinically used to treat large bone defects and complications such as delayed healing or nonunion. For the same dose of rhBMP-2, we have shown that a hybrid nanofiber mesh-alginate (NMA-rhBMP-2) delivery system provides longer-term release and increases functional bone regeneration in critically sized rat femoral bone defects compared with a collagen sponge. However, no comparisons of healing efficiencies have been made thus far between this hybrid delivery system and the gold standard of using autograft. Questions/purposes We compared the efficacy of the NMA-rhBMP-2 hybrid delivery system to morselized autograft and hypothesized that the functional regeneration of large bone defects observed with sustained BMP delivery would be at least comparable to autograft treatment as measured by total bone volume and ex vivo mechanical properties. Methods Bilateral critically sized femoral bone defects in rats were treated with either live autograft or with the NMA-rhBMP-2 hybrid delivery system such that each animal received one treatment per leg. Healing was monitored by radiography and histology at 2, 4, 8, and 12 weeks. Defects were evaluated for bone formation by longitudinal micro-CT scans over 12 weeks (n = 14 per group). The bone volume, bone density, and the total new bone formed beyond 2 weeks within the defect were calculated from micro-CT reconstructions and values compared for the 2-, 4-, 8-, and 12-week scans within and across the two treatment groups. Two animals were used for bone labeling with subcutaneously injected dyes at 4, 8, and 12 weeks followed by histology at 12 weeks to

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Section: Vital Bone Labelsmentioning

confidence: 99%

Hydrogel-based Delivery of rhBMP-2 Improves Healing of Large Bone Defects Compared With Autograft

Krishnan

Priddy

Esancy

et al. 2015

Clinical Orthopaedics &Amp; Related Research

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“…TECs were fixed in 4% paraformaldehyde for 1.5 h at 4°C, washed in 1· PBS for 10 min, preserved in 30% sucrose for 1 h, embedded in OCT media (Andwin Scientific, Addison, IL), and stored at -80°C. To prepare samples for IHC, TECs were cryosectioned longitudinally using cryofilm (Type 2C; Hiroshima, Japan) 45 at 150-200 mm from the top surface of the TECs. Sections were hydrated in 1· PBS, blocked (Power Block; Biogenix, Fremont, CA) for 30 min at room temperature (RT).…”

Section: Mp Fluorescence and Immunohistochemical Imagingmentioning

confidence: 99%

Fibrin Gels Exhibit Improved Biological, Structural, and Mechanical Properties Compared with Collagen Gels in Cell-Based Tendon Tissue-Engineered Constructs

Breidenbach

Dyment

et al. 2015

Tissue Engineering Part A

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The prevalence of tendon and ligament injuries and inadequacies of current treatments is driving the need for alternative strategies such as tissue engineering. Fibrin and collagen biopolymers have been popular materials for creating tissue-engineered constructs (TECs), as they exhibit advantages of biocompatibility and flexibility in construct design. Unfortunately, a few studies have directly compared these materials for tendon and ligament applications. Therefore, this study aims at determining how collagen versus fibrin hydrogels affect the biological, structural, and mechanical properties of TECs during formation in vitro. Our findings show that tendon and ligament progenitor cells seeded in fibrin constructs exhibit improved tenogenic gene expression patterns compared with their collagen-based counterparts for approximately 14 days in culture. Fibrin-based constructs also exhibit improved cell-derived collagen alignment, increased linear modulus (2.2-fold greater) compared with collagen-based constructs. Cyclic tensile loading, which promotes the maturation of tendon constructs in a previous work, exhibits a material-dependent effect in this study. Fibrin constructs show trending reductions in mechanical, biological, and structural properties, whereas collagen constructs only show improved tenogenic expression in the presence of mechanical stimulation. These findings highlight that components of the mechanical stimulus (e.g., strain amplitude or time of initiation) need to be tailored to the material and cell type. Given the improvements in tenogenic expression, extracellular matrix organization, and material properties during static culture, in vitro findings presented here suggest that fibrin-based constructs may be a more suitable alternative to collagen-based constructs for tissue-engineered tendon/ligament repair.

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“…To explore the possibility of chondrocyte to osteoblast transdifferentiation, we crossed Col10a1-mCherry mice with two different osteoblast reporter mouse lines, pOBCol3.6-Topaz and pOBCol2.3-Emerald (see Fig. 4), which have been extensively used to study osteoblast biology (BilicCurcic et al, 2005;Kalajzic et al, 2002Kalajzic et al, , 2005Ushiku et al, 2010). We reasoned that the persistent detection of Col10a1-mCherry fluorescence in cells present within the trabecular bone region may be used in a developmental fate mapping-like scheme and allow us to detect skeletal cells that express both chondrocyte and osteoblast reporters.…”

Section: Introductionmentioning

confidence: 99%

Generation and characterization of col10a1‐mcherry reporter mice

Maye

Butler

et al. 2011

Genesis

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SummaryWe report here on the generation of a new fluorescent protein reporter transgenic mouse line, Col10a1-mCherry, which can be used as a tool to study chondrocyte biology and pathology. Collagen, Type X, alpha 1 (Col10a1) is highly expressed in hypertrophic chondrocytes and commonly used as a gene marker for this cell population. The Col10a1-mCherry reporter line was generated using a bacterial recombination strategy with the mouse BAC clone RP23-192A7. To aid in the characterization of this animal model, we intercrossed Col10a1-mCherry mice with Collagen, Type II, alpha 1 (Col2a1) enhanced cyan fluorescent protein (ECFP) reporter mice and characterized the expression of both chondrocyte reporters during embryonic skeletal development from days E10.5 to E17.5. Additionally, at postnatal day 0, Col10a1-mCherry reporter expression was compared to endogenous Col10a1 mRNA expression in long bones and revealed that mCherry fluorescence extended past the Col10a1 expression domain. However, in situ hybridization for mCherry was consistent with the zone of Col10a1 mRNA expression, indicating that the persistent detection of mCherry fluorescence was a result of the long protein half life of mCherry in conjunction with a very rapid phase of skeletal growth and not due to aberrant transcriptional regulation. Taking advantage of the continued fluorescence of hypertrophic chondrocytes at the chondro-osseus junction, we intercrossed Col10a1-mCherry mice with two different Collagen, Type 1, alpha 1, (Col1a1) osteoblast reporter mice, pOBCol3.6-Topaz and pOBCol2.3-Emerald to investigate the possibility that hypertrophic chondrocytes transdifferentiate into osteoblasts. Evaluation of long bones at birth suggests that residual hypertrophic chondrocytes and osteoblasts in the trabecular zone exist as two completely distinct cell populations. genesis 49:410-418, 2011.

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Long bone fracture repair in mice harboring GFP reporters for cells within the osteoblastic lineage

Cited by 62 publications

References 47 publications

Hydrogel-based Delivery of rhBMP-2 Improves Healing of Large Bone Defects Compared With Autograft

Hydrogel-based Delivery of rhBMP-2 Improves Healing of Large Bone Defects Compared With Autograft

Fibrin Gels Exhibit Improved Biological, Structural, and Mechanical Properties Compared with Collagen Gels in Cell-Based Tendon Tissue-Engineered Constructs

Generation and characterization of col10a1‐mcherry reporter mice

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