The laboratory is composed of very skilled clinician scientists, basic science researchers, and biomedical engineers, which provides a multidisciplinary approach to address this complex disease process. To that effect, the lab is divided into six main areas of interest: gene- and cell-based therapy, biology of disc degeneration, biomarker discovery, mechanobiology, tissue engineering, and biomechanics.
- Gene and Cell Based Therapy
Current surgical intervention to treat intervertebral disc degeneration (IDD) addresses the resultant biomechanics of degeneration but neglects the underlying pathophysiology of disease. Gene and cell based therapies have the potential to address the imbalance between catabolism and anabolism that occurs within the disc tissue, thus potentially augmenting the course of IDD. The laboratory has continued to make progress in the area of gene and cell based therapy for the treatment of IDD.
Recent in vivo progress using gene therapy has yielded promising results. Using a rabbit model for degeneration, potentially therapeutic genes BMP-2 and TIMP-1 have been introduced to the intervertebral disc using an adeno-associated virus vector, resulting in delay of the degenerative cascade. Ongoing projects to better regulate TIMP-1 expression using novel response elements are being conducted in collaboration with Dr. Bing Wang.
- Biology of Disc Degeneration
The overall goal of Dr. Vo’s research is to understand the mechanisms of degeneration of matrix in the intervertebral discs and explore therapeutic strategies to minimize disc matrix loss. Our projects focus on investigating at the molecular and cellular levels how aging and tobacco smoking contribute to the loss of disc matrix. To achieve these goals, we collaborate with Dr. Laura Niedernhofer to use the novel progeroid mouse model (Ercc1-/∆ mice), which age rapidly due to a defect in repair of DNA damage. We also collaborate with Drs. Steven Shapiro, Peter Di, and Rocky Tuan to explore the mechanism of smoking-induced IDD. Dr. Sowa also continues to examine the potential benefit or detriment of commonly used oral supplements, such as glucosamine, chondroitin, and omega-3 fatty acids funded through NIH/NCCAM.
- Biomarker Discovery
Dr. Gwendolyn Sowa continues to investigate how serum concentrations of certain biomarkers correlate with the presence and severity of IDD. Current interests include examining potential superiority to imaging based findings in predicting pain and pain related disability, as well as response to activity and other treatment modalities. New avenues of research include utilizing serum-based biomarkers in conjunction with imaging biomarkers to predict individual responses to treatments, such as interventional spine procedures.
Dr. Sowa also continues to explore the effects of mechanical loading in a novel ex vivo intervertebral disc testing system, which maintains biologic activity. Dr. Robert Tisherman is working on extending the capabilities of the system to include small animal models. Our goal is to gain a better understanding of the biochemical processes that occur with loading to inform future motion based therapies to facilitate healing and/or regeneration of the intervertebral disc.
- Facilities and Resources
The Ferguson Laboratory has approximately 3,500 square feet devoted to biochemistry, histology, tissue culture, and biomechanical testing. In addition, the lab’s faculty, staff, and trainees have 1,000 square feet of office space. Our team also collaborates with researchers in other labs—the Department of Orthopedics and Physical Medicine and Rehabilitation, including the Mechanobiology Laboratory, and Human Engineering Research Laboratory. The University of Pittsburgh’s Center for Biological Imaging, UPMC Investigational Drug Service, and Magnetic Resonance Research Center are also utilized as needed.
- Wet Lab
The general wet lab area facilitates protein analysis, gene expression, and various ELISAs. It includes the following: one chemical fume hood, standard air, vacuum, gas, and water provided to bench work stations, three bio safety cabinets, Bio-Rad polyacrylamide slab gel electrophoresis equipment and accessories, agarose gel electrophoresis facilities, three benchtop centrifuges, Perkin Elmer plate-reading spectrophotometer, PCR Workstation, Bio-Rad iQ5 PCR system, and gramatic and microgramatic balances.
The histology section of the lab houses two cutting workstations, a cryostat and tissue preparation stations, and a paraffin embedder. This allows the lab to utilize immunohistochemistry (IHC), immunofluorescence (IF), and other staining techniques. A microscopy room includes a Nikon Eclipse TS100 inverted microscope, a Nikon Eclipse E800 fluorescent transmission microscope, a stereo microscope for dissection, and an RTse SPOT camera. Visualization of IHC or IF can be accomplished within the lab, but expert support also exists from the university in the form of its core imaging facility, the Center for Biological Imaging.
- Tissue Culture
The tissue culture space consists of three sterile hoods, four HERACell 150i’s, and one ThermoForma Series II Water Jacketed CO2 incubator. Tissue culture facilities enable cell and organ culture.
The biomechanics region holds a Staubli RX90 serial-linkage robot, NDI Polaris motion capture system, a 5-camera Vicon motion capture system, a benchtop axial testing machine, a dissolved oxygen probe, 4-wire RTD temperature sensor, BK Precision programmable power supply, a HERACell 150i with 1-21% O2 capability, standing drill press, band saw, and work bench. These tools are helpful for prototype design and modification of existing fixtures, but design and fabrication are supported by machine and electronic shops within the School of Medicine and the School of Engineering.