Thomas E. Patterson

Cellular Strategies for Enhancement of Fracture Repair

Tissue engineering seeks to translate scientific knowledge into tangible products to advance the repair, replacement, or regeneration of organs and tissues. Current tissue engineering strategies have progressed recently from a historical approach that is based primarily on biomaterials to a cell and tissue-based approach that includes understanding of cell-sourcing and bioactive stimuli. New options include methods for harvest and transplantation of tissue-forming cells, bioactive matrix materials that act as tissue scaffolds, and delivery of bioactive molecules within scaffolds. These strategies are already benefiting patients, and they place increasing demands on orthopaedic surgeons to have a solid foundation in the contemporary concepts and principles of cell-based tissue engineering. Essentially all orthopaedic tissue engineering strategies can be distilled to a strategy or combination of strategies that seek to increase the number or relative performance of bone-forming cells. The global term connective tissue progenitors has been used to define the heterogeneous populations of stem and progenitor cells that are found in native tissue and that are capable of differentiating into one or more connective tissue phenotypes. These stem or progenitor populations are found in various tissue sources, with varying degrees of ability to differentiate along connective tissue lineages. Available cell-based strategies include targeting local cells with use of scaffolds or bioactive factors, or transplantation of autogenous connective tissue progenitor cells derived from bone marrow or other tissues, with or without processing to change their concentration or prevalence. The future may include means of homing circulating connective tissue progenitor cells with use of intrinsic chemokine systems, or modifying the biological performance of connective tissue progenitor cells by means of genetic modifications.

Three-Dimensional Imaging and Templating Improve Glenoid Implant Positioning

BACKGROUND Preoperative quantitative assessment of glenoid bone loss, selection of the glenoid component, and definition of its desired location can be challenging. Placement of the glenoid component in the desired location at the time of surgery is difficult, especially with severe glenoid pathological conditions. METHODS Forty-six patients were randomly assigned to three-dimensional computed tomographic preoperative templating with either standard instrumentation or with patient-specific instrumentation and were compared with a nonrandomized group of seventeen patients with two-dimensional imaging and standard instrumentation used as historical controls. All patients had postoperative three-dimensional computed tomographic metal artifact reduction imaging to measure and to compare implant position with the preoperative plan. RESULTS Using three-dimensional imaging and templating with or without patient-specific instrumentation, there was a significant improvement achieving the desired implant position within 5° of inclination or 10° of version when compared with two-dimensional imaging and standard instrumentation. CONCLUSION Three-dimensional assessment of glenoid anatomy and implant templating and the use of these images at the time of surgery improve the surgeons ability to place the glenoid implant in the desired location.

Magnetic Field Visualization in Applications to Pulsed Electromagnetic Field Stimulation of Tissues

AbstractElectromagnetic field visualization is important in multidisciplinary research on the molecular basis of therapeutic effects of pulsed electromagnetic fields (PEMF). We have compared classic PEMF representations by two-dimensional field lines and field magnitude contour plots with a field representation using three-dimensional field isosurfaces. Field simulations were performed for a clinically approved Spinal-Stim® Lite system (Orthofix Inc., McKinney, TX). The relatively simple coil system geometry and the predominantly dielectric properties of the surrounding medium (air and human connective tissue) allowed us to develop analytical expressions for the field. The field model was validated by comparison with experimentally measured field values, and with values calculated using a commercial finite-element analysis software package. Two-dimensional field representations by field lines and field contour plots were less intuitive than three-dimensional field isosurface representations to members of the group without an engineering background. Field isosurfaces, represented as three-dimensional solids, allowed for direct visualization of PEMF targeting of individual organs (lumbar spine), the extent of the therapeutic field value, and the directional field characteristics. The dynamic characteristic of the field was well illustrated by a sequence of field isosurfaces corresponding to the evolution with time of the electric current waveform (sawtooth) powering the coils. The isosurface representation of the field can be extended to any three-dimensional coil system geometry using plotting capabilities of current computer algebra software packages.

Quantitative Measurement of Osseous Pathology in Advanced Glenohumeral Osteoarthritis

Background: Osteoarthritis of the glenohumeral joint has typical patterns of deformity as described by Walch et al. However, more severe glenoid pathology may be difficult to classify. The purpose of this study was to use 3-dimensional computed tomography (3-D CT) imaging analysis to define common pathologic subtypes that can be differentiated from the current Walch classification. Methods: We performed quantitative measurements of premorbid and pathologic anatomy using preoperative 3-D CT scans from 155 cases of advanced glenohumeral osteoarthritis that underwent anatomic or reverse total shoulder arthroplasty. We defined premorbid glenohumeral anatomy on the basis of previously validated methods using 3-D glenoid vault and humeral best-fit circle models including the premorbid glenoid version, joint-line medialization, and humeral-glenoid alignment (HGA). We determined the anatomic features that differentiate new glenoid morphologic subtypes from the existing Walch classification both qualitatively and quantitatively. Results: We defined 2 new glenoid subtypes (B3 and C2) for which the glenoid pathology and humeral alignment were not defined in the original Walch classification. The B3 glenoid has high pathologic retroversion, normal premorbid version, and acquired central and posterior bone loss that, on average, is greater than that of the B2 glenoid. The C2 glenoid is dysplastic with high pathologic retroversion, high premorbid version, and acquired posterior bone loss, giving it the appearance of a biconcave glenoid with posterior translation of the humeral head. This C2 glenoid can be confused with the B2 glenoid. Conclusions: The B3 and C2 patterns have qualitative and quantitative differences that may result in different clinical outcomes than classic B2 or C types; therefore, our findings suggest that these new subtypes should be included in a new or modified classification system.

Slowing the Onset of Hypoxia Increases Colony Forming Efficiency of Connective Tissue Progenitor Cells In Vitro.

BACKGROUND Survival and colony formation by transplanted tissue derived connective tissue progenitor cells (CTPs) are thought to be important factors in the success of clinical tissue engineering strategies for bone regeneration. Transplantation of cells into defects larger than a few millimeters expose cells to a profoundly hypoxic environment. This study tested the hypothesis that delaying the onset of hypoxia will improve the survival and performance of CTPs in vitro. METHODS To mimic declines seen in an avascular in vivo bone defect, colony forming efficiency by marrow derived nucleated cells was assessed under osteogenic conditions. Variation in the rate of oxygen decline from an oxygen tension of 21% to 0.1% oxygen was explored using an incubator with programmable active control of gas concentrations. The effect of doping cultures with defined concentrations of RBCs was also used to evaluate the potential for RBCs to serve as a natural buffer in the setting of declining oxygen levels. RESULTS A delay in onset of hypoxia over 96 hours resulted in a 3-fold increase in the relative colony forming efficiency (rCFE) of CTPs as compared to an immediate onset of hypoxia. The presence of RBCs in vitro inhibited the rCFE of CTPs. Given the negative effects of RBCs, methods of RBC removal were evaluated and compared for their effectiveness of RBC removal and retention of colony forming efficiency. CONCLUSIONS These data suggest that conditions of hypoxia compromise colony forming efficiency in marrow derived CTPs. However, slowing the rate of decline of oxygen preserved colony forming efficiency at levels achieved in a stable normoxic (3% O2) environment. These data also suggest that RBCs are detrimental to the rCFE of CTPs and that buffy coat is an effective and preferred method for removing RBCs from marrow aspirates while preserving CTPs. These findings may inform clinical strategies for CTP transplantation.

The Efficiency of Bone Marrow Aspiration for the Harvest of Connective Tissue Progenitors from the Human Iliac Crest

Background: The rational design and optimization of tissue engineering strategies for cell-based therapy requires a baseline understanding of the concentration and prevalence of osteogenic progenitor cell populations in the source tissues. The aim of this study was to (1) define the efficiency of, and variation among individuals in, bone marrow aspiration as a means of osteogenic connective tissue progenitor (CTP-O) harvest compared with harvest from iliac cancellous bone, and (2) determine the location of CTP-Os within native cancellous bone and their distribution between the marrow-space and trabecular-surface tissue compartments. Methods: Eight 2-mL bone marrow aspiration (BMA) samples and one 7-mm transcortical biopsy sample were obtained from the anterior iliac crest of 33 human subjects. Two cell populations were obtained from the iliac cancellous bone (ICB) sample. The ICB sample was placed into &agr;MEM (alpha-minimal essential medium) with antibiotic-antimycotic and minced into small pieces (1 to 2 mm in diameter) with a sharp osteotome. Cells that could be mechanically disassociated from the ICB sample were defined as marrow-space (IC-MS) cells, and cells that were disassociated only after enzymatic digestion were defined as trabecular-surface (IC-TS) cells. The 3 sources of bone and marrow-derived cells were compared on the basis of cellularity and the concentration and prevalence of CTP-Os through colony-forming unit (CFU) analysis. Results: Large variation was seen among patients with respect to cell and CTP-O yield from the IC-MS, IC-TS, and BMA samples and in the relative distribution of CTP-Os between the IC-MS and IC-TS fractions. The CTP-O prevalence was highest in the IC-TS fraction, which was 11.4-fold greater than in the IC-MS fraction (p < 0.0001) and 1.7-fold greater than in the BMA fraction. However, the median concentration of CTP-Os in the ICB (combining MS and TS fractions) was only 3.04 ± 1.1-fold greater than that in BMA (4,265 compared with 1,402 CTP/mL; p = 0.00004). Conclusions: Bone marrow aspiration of a 2-mL volume at a given needle site is an effective means of harvesting CTP-Os, albeit diluted with peripheral blood. However, the median concentration of CTP-Os is 3-fold less than from native iliac cancellous bone. The distribution of CTP-Os between the IC-MS and IC-TS fractions varies widely among patients. Clinical Relevance: Bone marrow aspiration is an effective means of harvesting CTP-Os but is associated with dilution with peripheral blood. Overall, we found that 63.5% of all CTP-Os within iliac cancellous bone resided on the trabecular surface; however, 48% of the patients had more CTP-Os contributed by the IC-MS than the IC-TS fraction.