Research Capabilities

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.

• 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

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

• 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