Research Capabilities

The Orthopaedic Research Center is a comprehensive research facility predominantly focusing on the prevention and repair of ortheopaedic disease in human and animals. In addition to protein biomarker analysis and development, this program is supported by several molecular biology applications such as antibody purification, real time PCR assay development and gene expression analysis, cell and tissue culture techniques, adenoviral construction and cloning, gene chip microarray, biomechanical testing, and histological procedures.

As the support structure for biomedical research continues to expand with modern medical discoveries and advances, the Orthopaedic Research Center will continue to provide groundbreaking research for the future.

Research techniques

Biomarker Analysis

Fully equipped to run any commercially available absorbance or fluorescence biomarker immunoassay in a 96-well plate format, using Molecular Devices SpectraMax, microplane absorbance/transmittance reader, as well as a Gemini-XS Fluorometer.

Extensive experience with the following biomarker assays:

Detection of Cartilage Markers

  • Alcian Blue: Standardize measurement of 35S labeled proteoglycan complexes
  • C1, C2: An assay to standardize the measurement of Types I and II collagen degradation
  • CPII: An assay to measure type II collagen carboxy propeptide (C-propeptide).
  • CS-846: Measurement of Aggrecan Chondroitin Sulfate 846 Epitope.
  • Eq. Col 2. (CEQ): An assay to quantify equine specific Type II collagen, which has also been proven to work with canine fluid.
  • GAG DMMB: An assay ¬†for standardized measurement of glycosaminoglycans in biological fluids and/or tissues.
  • Pyd Assay: An assay to standardize measurement of pyridinoline crosslinks in serum and urine.
  • Pyrilinks-D: To standardize measurements of deoxypyridinoline crosslinks in urine.
  • TCA: Assay to measure 3H content in media or cartilage digested samples
  • YKL-40: Assay for measurement of YKL-40 human cartilage glycoprotein 39, in serum
  • Sircol Assay: Assay to assess the amount of newly synthesized collagen in cartilage, tendon or cell culture media.

Detection of Bone Markers

  • C1,2C: An assay to standardize measurement of Type I and II collagens (378 assay, MMP1 and MMP13).
  • Metra(tm) BAP: Quantification of bone-specific alkaline phosphatase in serum and synovial fluid samples.
  • Metra(tm) Osteocalcin EIA: An enzyme immunoassay for the quantification of intact (de novo) osteocalcin.
  • Serum Cross Laps(registered) (CTX): Assay for the quantification of degradation products of C-terminal telopeptides of Type-I collagen in serum and plasma.

Pre-Assay Sample Processing

Includes papain, hyaluronidase, and collogenase digestion, as well as chromatography extraction of synovial fluid, serum, and tissues.

Western, Southern, and Northern Blotting

Other

  • PDGF-BB: An assay to quantify levels of Platelet-Derived Growth Factor-BB subunit in serum, plasma, and cell culture supernatant.
  • PGE2: An assay to quantify levels of Prostaglandin E2 in serum, plasma, synovial fluid, cell culture supernatant, and urine.

Many other assays available – please inquire.

Biomechanical Testing

  • Displacement control testing for compressive, tension, and shear material properties
  • Tissue explants or cell-seeded scaffolds
  • Light to moderate load cells are suitable for resting small tissue explants or cell-seeded scaffolds

Molecular Biology

Evaluation of Metabolic Activity in Living Tissue

Radiolabel protocols available

Real Time PCR Analysis

  • ABI Prism(registered) 7000 Sequence Detection System
  • Optimization of PCR Primers

RNA/DNA Extractions/Isolations

  • cDNA synthesis from RNA
  • RNA from cells, tissues, or whole blood
  • Primer and probe design
  • Gel extraction and purification
  • Purification of plasmid DNA
  • PCR amplification

Isolation of Synoviocytes, Chondrocytes, and Tenocytes

Cell culture expansion of freshly collected cells

Culturing of Mesenchymal Stem Cells (bone-marrow derived or fat-derived)

  • Cell culture expansion of bone-marrow derived or adipose-derived cells, including three dimensional culturing for clinical use
  • Flow cytometry analysis of MSC and other cellular cultures

Adenovial Vector Construction and Cell Transfection

The development of adenovial vectors for the delivery of genes into cells

Histology Services

  • Decalcified tissue histology
  • Immunohistochemistry
  • Paraffin and fresh frozen sectioning and staining of paraffin embedded samples
  • Live/Dead cellular tissue staining and fluorescent imaging
  • Histomorphometric analysis

Current concepts

Equine Performance Analysis Facility

Learn more about our capabilities at the Equine Performance Analysis Facility.