Iliac dysmorphism: defining radiographic characteristics and association with pelvic osseous corridor size.

INTRODUCTION: Insertion of iliac wing implants requires understanding of the curvilinear shape of the ilium. This study serves to quantitatively identify the area of iliac inner-outer table convergence (IOTC), characterize the iliac wing osseous corridor, and define the gluteal pillar osseous corridor. METHODS: Computed tomography scans of 100 male and 100 female hemipelves were evaluated. The iliac wing was studied using manual best-fit analysis of the bounds of the inner and outer cortices. The IOTC was defined as the location of the iliac wing with an intercortical width less than 5 mm. The shortest distance from the apex of the iliac crest to the superior border of the IOTC was defined as the iliac wing osseous corridor. Finally, the width of the gluteal pillar corridor from the gluteus medius tubercle to the ischial tuberosity was measured. RESULTS: The IOTC is an elliptical area measuring 22.3 cm2. All ilia had an area where the inner and outer cortices converged to an intercortical width of less than 5 mm; 48% converged to a single cortex. The shortest mean distance from the superior edge of the iliac crest to the beginning of the IOTC was 20.3 mm in men and 13.8 mm in women (p < 0.001). The gluteal pillar diameter averaged 5.3 mm in men and 4.3 mm in women (p < 0.001). DISCUSSION: All ilia converge to a thin and frequently unicortical central region. A 4.5 mm iliac wing lag screw will not breach the cortex if it remains within 20 mm or 14 mm distal to the cranial aspect of the iliac crest in males and females, respectively. Not only is the gluteal pillar smaller than previously thought, in 41% of males and 73% of females, it is not be large enough for 5 mm implants. CONCLUSION: This study quantitatively assesses the dimensions of the IOTC, the iliac crest osseous corridor, and the gluteal pillar. Overall, our findings provide improved understanding of the limits for implant use in the iliac wing as well as better appreciation of the complex osteology of the ilium. This will help surgeons to identify safe areas for implant placement and avoid inadvertent cortical penetration.

Defining the iliac wing osseous fixation pathways: anatomy and implant constriction points.

INTRODUCTION: Insertion of iliac wing implants requires understanding of the curvilinear shape of the ilium. This study serves to quantitatively identify the area of iliac inner-outer table convergence (IOTC), characterize the iliac wing osseous corridor, and define the gluteal pillar osseous corridor. METHODS: Computed tomography scans of 100 male and 100 female hemipelves were evaluated. The iliac wing was studied using manual best-fit analysis of the bounds of the inner and outer cortices. The IOTC was defined as the location of the iliac wing with an intercortical width less than 5 mm. The shortest distance from the apex of the iliac crest to the superior border of the IOTC was defined as the iliac wing osseous corridor. Finally, the width of the gluteal pillar corridor from the gluteus medius tubercle to the ischial tuberosity was measured. RESULTS: The IOTC is an elliptical area measuring 22.3 cm2. All ilia had an area where the inner and outer cortices converged to an intercortical width of less than 5 mm; 48% converged to a single cortex. The shortest mean distance from the superior edge of the iliac crest to the beginning of the IOTC was 20.3 mm in men and 13.8 mm in women (p < 0.001). The gluteal pillar diameter averaged 5.3 mm in men and 4.3 mm in women (p < 0.001). DISCUSSION: All ilia converge to a thin and frequently unicortical central region. A 4.5 mm iliac wing lag screw will not breach the cortex if it remains within 20 mm or 14 mm distal to the cranial aspect of the iliac crest in males and females, respectively. Not only is the gluteal pillar smaller than previously thought, in 41% of males and 73% of females, it is not be large enough for 5 mm implants. CONCLUSION: This study quantitatively assesses the dimensions of the IOTC, the iliac crest osseous corridor, and the gluteal pillar. Overall, our findings provide improved understanding of the limits for implant use in the iliac wing as well as better appreciation of the complex osteology of the ilium. This will help surgeons to identify safe areas for implant placement and avoid inadvertent cortical penetration.

Supraacetabular osseous corridor: defining dimensions, sex differences, and alternatives.

INTRODUCTION: The supraacetabular (SA) corridor extends from the anterior inferior iliac spine to the posterior ilium and can safely accommodate implants to stabilize pelvic and acetabular fractures. However, quantitative analysis of its dimensions and characteristics have not been thoroughly described. This study seeks to define the dimensions, common constriction points, and any alternative trajectories that would maximize the corridor diameter. METHODS: Computed tomography of 100 male and 100 female hemipelves without osseous trauma were evaluated. The corridor boundaries were determined through manual best-fit analysis. The largest intercortical cylinder within the pathway was created and measured. Alternative trajectories were tested within the SA boundaries to identify another orientation that maximized the diameter of the intercortical cylinder. RESULTS: The traditional SA corridor had a mean diameter of 8.3 mm in men and 6.2 mm in women. This difference in diameter is due to a more S-shaped ilium in women. A larger alternative SA corridor was found that had a less limited path through the ilium and measured 11.3 mm in men and 9.9 mm in women. These dimensions are significantly different compared to those of the traditional SA corridor in both men and women. CONCLUSIONS: In men, the SA corridor allows for the safe passage of most hardware used in pelvic and acetabular fractures. However, in women, the SA corridor is restricted by a more S-shaped ilium. An alternative trajectory was found that has a significantly larger mean diameter in both sexes. Ultimately, the trajectory of hardware will be dictated by the clinical scenario. When large implants are needed, especially in women, we recommend considering the alternative SA corridor.

Promoting Endochondral Bone Repair Using Human Osteoarthritic Articular Chondrocytes.

INTRODUCTION: Current tissue engineering strategies to heal critical-size bone defects through direct bone formation are limited by incomplete integration of grafts with host bone and incomplete graft vascularization. An alternative strategy for bone regeneration is the use of cartilage grafts that form bone through endochondral ossification. Endochondral cartilages stimulate angiogenesis and are remodeled into bone, but are found in very small quantities in growth plates and healing fractures. We sought to develop engineered endochondral cartilage grafts using osteoarthritic (OA) articular chondrocytes as a cell source. Such chondrocytes often undergo hypertrophy, which is a characteristic of endochondral cartilages. MATERIALS AND METHODS: We compared the ability of unmodified human OA (hOA) cartilage and cartilage grafts formed in vitro from hOA chondrocytes to undergo endochondral ossification in mice. Scaffold-free engineered chondrocyte grafts were generated by pelleting chondrocytes, followed by culture with transforming growth factor-ß1 (TGF-ß1) and bone morphogenetic protein 4. Samples derived from either primary or passaged chondrocytes were implanted subcutaneously into immunocompromised mice. Grafts derived from passaged chondrocytes from three patients were implanted into critical-size tibial defects in mice. Bone formation was assessed with histology after 4 weeks of implantation. The composition of tibial repair tissue was quantified with histomorphometry. RESULTS: Engineered cartilage grafts generated from passaged OA chondrocytes underwent endochondral ossification after implantation either subcutaneously or in bone. Cartilage grafts integrated with host bone at 15 out of 16 junctions. Grafts variably remodeled into woven bone, with the proportion of bony repair tissue in tibial defects ranging from 22% to 85% (average 48%). Bony repair tissue bridged the tibial defects in half of the animals. In contrast, unmodified OA cartilage and engineered grafts formed from primary chondrocytes did not undergo endochondral ossification in vivo. CONCLUSIONS: hOA chondrocytes can adopt an endochondral phenotype after passaging and TGF-ß superfamily treatment. Engineered endochondral cartilage grafts can integrate with host bone, undergo ossification, and heal critical-size long-bone defects in a mouse model. However, additional methods to further enhance ossification of these grafts are required before the clinical translation of this approach.

Trophic stimulation of articular chondrocytes by late-passage mesenchymal stem cells in coculture.

Coculture of mesenchymal stem cells (MSCs) with articular chondrocytes (ACs) increases glycosaminoglycan (GAG) accumulation compared to monoculture. MSCs might (1) differentiate into ACs (progenitor role) and/or (2) stimulate AC matrix metabolism (trophic role). MSCs lose the ability to undergo chondrogenesis after extended passaging. We hypothesized that MSCs act predominantly as progenitors, and that late-passage MSCs without chondrogenic potential would be unable to increase GAG in coculture. Early- and late-passage human MSCs (hMSCs) were grown in pellet monoculture under chondrogenic conditions and in pellet coculture with bovine ACs. Chondrogenesis was assessed with GAG quantification, safranin-O staining, and quantitative PCR (qPCR). Contributions of human and bovine cells were assessed with species-specific qPCR and human-specific immunostaining. Late-passage hMSCs did not undergo chondrogenesis in monoculture with chondrogenic stimuli or in coculture with ACs. Early-passage hMSCs underwent chondrogenesis only in response to chondrogenic stimuli. Coculture pellets in both cases accumulated as much GAG/DNA as monoculture AC pellets. Aggrecan transcription was not increased in coculture. Late-passage hMSCs that do not undergo chondrogenesis are capable of stimulating GAG accumulation in coculture with ACs. This supports a trophic effect of hMSCs on ACs. hMSCs may have therapeutic utility even after prolonged passaging.

Cytotoxicity of local anesthetics on human mesenchymal stem cells.

BACKGROUND: Local anesthetics are frequently delivered intra-articularly to provide perioperative pain control. Previous studies have shown that the commonly used drugs lidocaine, ropivacaine, and bupivacaine can be toxic to human chondrocytes. The present study was conducted to determine whether the toxic effects of local anesthetics on human chondrocytes also extend to human mesenchymal stem cells. METHODS: Human mesenchymal stem cells from three healthy donors were grown in tissue culture and exposed to the following anesthetic treatments for sixty minutes: (1) 1% lidocaine, (2) 2% lidocaine, (3) 0.25% bupivacaine, (4) 0.5% bupivacaine, (5) 0.2% ropivacaine, and (6) 0.5% ropivacaine. The cells were then allowed to recover for twenty-four hours in regular growth media, and viability was measured with use of fluorescent staining for live cells or a luminescence assay for ATP content. RESULTS: The live cell counts and ATP content were correlated (r2 = 0.79), and 2% lidocaine was found to be significantly more toxic than all doses of bupivacaine and ropivacaine. Treatment with 1% lidocaine resulted in significantly fewer live cells (49%) compared with the control, and the live cell count was also significantly less than that for the other anesthetics. However, the ATP level in the 1% lidocaine group was not significantly lower than those in the other groups. Bupivacaine and ropivacaine did not exhibit significant differences in toxicity compared with the control or with each other. CONCLUSIONS: Ropivacaine and bupivacaine had limited toxicity in human mesenchymal stem cells. However, lidocaine could significantly decrease mesenchymal stem cell viability. Since other studies have shown ropivacaine to be less toxic to chondrocytes than bupivacaine, ropivacaine may be a safer intra-articular anesthetic. CLINICAL RELEVANCE: Mesenchymal stem cells likely play a key role in healing following surgical procedures such as microfracture and ligament reconstruction. If local anesthetics are used following joint surgery, selection of an agent with low toxicity toward mesenchymal stem cells, such as ropivacaine, may maximize tissue healing potential.

Inducible immortality in hTERT-human mesenchymal stem cells.

Human mesenchymal stem cells (hMSCs) are attractive candidates for tissue engineering and cell-based therapy because of their multipotentiality and availability in adult donors. However, in vitro expansion and differentiation of these cells is limited by replicative senescence. The proliferative capacity of hMSCs can be enhanced by ectopic expression of telomerase, allowing for long-term culture. However, hMSCs with constitutive telomerase expression demonstrate unregulated growth and even tumor formation. To address this problem, we used an inducible Tet-On gene expression system to create hMSCs in which ectopic telomerase expression can be induced selectively by the addition of doxycycline (i-hTERT hMSCs). i-hTERT hMSCs have inducible hTERT expression and telomerase activity, and are able to proliferate significantly longer than wild type hMSCs when hTERT expression is induced. They stop proliferating when hTERT expression is turned off and can be rescued when expression is re-induced. They retain multipotentiality in vitro even at an advanced age. We also used a selective inhibitor of telomere elongation to show that the mechanism driving immortalization of hMSCs by hTERT is dependent upon maintenance of telomere length. Thanks to their extended lifespan, preserved multipotentiality and controlled growth, i-hTERT hMSCs may prove to be a useful tool for the development and testing of novel stem cell therapies.

Nucleic acid recognition by Toll-like receptors is coupled to stepwise processing by cathepsins and asparagine endopeptidase.

Toll-like receptor (TLR) 9 requires proteolytic processing in the endolysosome to initiate signaling in response to DNA. However, recent studies conflict as to which proteases are required for receptor cleavage. We show that TLR9 proteolysis is a multistep process. The first step removes the majority of the ectodomain and can be performed by asparagine endopeptidase (AEP) or cathepsin family members. This initial cleavage event is followed by a trimming event that is solely cathepsin mediated and required for optimal receptor signaling. This dual requirement for AEP and cathepsins is observed in all cell types that we have analyzed, including mouse macrophages and dendritic cells. In addition, we show that TLR7 and TLR3 are processed in an analogous manner. These results define the core proteolytic steps required for TLR9 function and suggest that receptor proteolysis may represent a general regulatory strategy for all TLRs involved in nucleic acid recognition.