Determination of variations in gene expression during fracture healing

Balaburski, Gregor M.; O’Connor, J. P.

doi:10.1080/00016470310013608

Cited by 13 publications

(5 citation statements)

References 36 publications

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“…Total RNA was isolated from fracture calluses at 4 and 7 days post‐fracture and cDNA was obtained by reverse transcription as described previously 12. Target cDNAs were quantified by real‐time PCR using SYBR Green I fluorescence as measure of DNA amplification in 25 µl reactions (ABgene, Rochester, NY).…”

Section: Methodsmentioning

confidence: 99%

Effects of local insulin delivery on subperiosteal angiogenesis and mineralized tissue formation during fracture healing

Paglia¹,

Wey²,

Breitbart³

et al. 2012

Journal Orthopaedic Research

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Local insulin delivery has been shown to improve osseous healing in diabetic animals. The purpose of this study was to quantify the effects of local intramedullary delivery of saline or Ultralente insulin (UL) on various fracture healing parameters using an in vivo non-diabetic BB Wistar rat model. Quantitation of local insulin levels showed a rapid release of insulin from the fractured femora, demonstrating complete release at 2 days. RT-PCR analysis revealed that the expression of early osteogenic markers (Col1a2, osteopontin) was significantly enhanced with UL treatment when compared with saline controls (p < 0.05). Significant differences in VEGF þ cells and vascularity were evident between the treatment and control groups at day 7 (p < 0.05). At day 21, histomorphometric analysis demonstrated a significant increase in percent mineralized tissue in the UL-treated animals compared with controls (p < 0.05), particularly within the subperiosteal region of the fracture callus. Mechanical testing at 4 weeks showed significantly greater mechanical strength for UL-treated animals (p < 0.05), but healing in control animals caught up at 6 weeks post-fracture. These results suggest that the primary osteogenic effect of UL during the early stages of fracture healing (1-3 weeks) is through an increase in osteogenic gene expression, subperiosteal angiogenesis, and mineralized tissue formation. ß

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Section: Methodsmentioning

confidence: 99%

Effects of local insulin delivery on subperiosteal angiogenesis and mineralized tissue formation during fracture healing

Paglia¹,

Wey²,

Breitbart³

et al. 2012

Journal Orthopaedic Research

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“…The RNA was further purified by differential binding and elution from glass fibers. Total RNA was purified from individual calluses as described previously 33. cDNA was prepared from 1 µg total RNA in a 20 µl RT reaction.…”

Section: Methodsmentioning

confidence: 99%

The effects of low‐intensity pulsed ultrasound upon diabetic fracture healing

Coords

Breitbart

Paglia

et al. 2010

Journal Orthopaedic Research

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ABSTRACT:In the United States, over 17 million people are diagnosed with type 1 diabetes mellitus (DM) with its inherent morbidity of delayed bone healing and nonunion. Recent studies demonstrate the utility of pulsed low-intensity ultrasound (LIPUS) to facilitate fracture healing. The current study evaluated the effects of daily application of LIPUS on mid-diaphyseal femoral fracture growth factor expression, cartilage formation, and neovascularization in DM and non-DM BB Wistar rats. Polymerase chain reaction (PCR) and ELISA assays were used to measure and quantify growth factor expression. Histomorphometry assessed cartilage formation while immunohistochemical staining for PECAM evaluated neovascularization at the fracture site. In accordance with previous studies, LIPUS was shown to increase growth factor expression and cartilage formation. Our study also demonstrated an increase in fracture callus neovascularization with the addition of LIPUS. The DM group showed impaired growth factor expression, cartilage formation, and neovascularization. However, the addition of LIPUS significantly increased all parameters so that the DM group resembled that of the non-DM group. These findings suggest a potential role of LIPUS as an adjunct for DM fracture treatment.

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“…During mechanical loading, mechano-sensitive genes are up-or down-regulated and their expression patterns have been shown to affect cell activity rates in the course of tissue formation Barry et al, 2001;Kelly and Jacobs, 2010;Nowlan et al, 2008;Palomares et al, 2009;Provot and Schipani, 2005). In addition, mechano-sensitive genes are expressed with a large variation in animals of the same species compared to housekeeping genes (Balaburski and O'Connor, 2003). It has also been shown that mechano-sensitivity is positively correlated with bone turnover rates and negatively associated with bone mineral density (Judex et al, 2002), which in turn affects the course of tissue differentiation and fracture healing (Balaburski and O'Connor, 2003;Jepsen et al, 2008;Li et al, 2001;Manigrasso and O'Connor, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…In addition, mechano-sensitive genes are expressed with a large variation in animals of the same species compared to housekeeping genes (Balaburski and O'Connor, 2003). It has also been shown that mechano-sensitivity is positively correlated with bone turnover rates and negatively associated with bone mineral density (Judex et al, 2002), which in turn affects the course of tissue differentiation and fracture healing (Balaburski and O'Connor, 2003;Jepsen et al, 2008;Li et al, 2001;Manigrasso and O'Connor, 2008). It is therefore intriguing to wonder if variation in mechano-sensitivity can explain the variation in tissue differentiation observed in vivo.…”

Section: Introductionmentioning

confidence: 99%