Hanspeter Frei

The effect of Nucleotomy on lumbar spine mechanics in compression and shear loading

Study Design. An in vitro biomechanical investigation on human cadaveric specimens was conducted before and after nucleotomy. Endplate and vertebral body deformation patterns were measured under compression and shear loading, in addition to kinematics and disc pressure. Objective. The working hypotheses of this study were that in compression, nucleotomy results in an altered deformation pattern of the endplate and that in shear, nucleotomy does not result in an altered endplate deformation pattern or disc pressure. Summary of Background Data. The pressure distributions within the intervertebral disc have been studied in compression loading but not in shear loading. Severe degeneration and surgical nucleotomy result in small nuclear pressure and altered loading distribution in compression. The effect of these changes on the vertebral endplate and the response under shear loads are not well understood. Methods. Five L3–L4 and two L4–L5 functional spinal units were tested under compression and shear loading, intact and after nucleotomy. Vertebral body deformations, intradiscal pressure, and intervertebral kinematics were measured. A series of compression-type (maximum 1000 N) and shear-type (maximum 500 N) loads were applied. Results. With nucleotomy, the disc pressure and the endplate strains decreased under compression, but the vertebral rim strains did not change. In shear, the vertebral rim and endplate strains did not change with nucleotomy. Disc pressure was lower in shear than in compression. Conclusion. Nucleotomy resulted in decreased disc pressure, decreased endplate deformation, and modified loading patterns onto the inferior vertebra in compression loading. However, nucleotomy did not appreciably affect the behavior of the disc in shear loading.

Thoracolumbar spine mechanics contrasted under compression and shear loading

The mechanical properties of the human spine have been studied extensively in compression, but there remains a lack of fundamental data in shear. The overall goal of this study was to contrast the mechanics of the thoracolumbar functional spinal unit (FSU) under compression and shear‐type loads by evaluating endplate deformation, disc pressures, and kinematics between the different loading types. Eleven T12‐L1 and one L1‐L2 human FSUs were tested. Compression loads consisted of pure compression, extension‐compression, flexion‐compression, lateral left and right compression applied individually to a maximum of 500 N. Shear loading consisted of posterior, anterior, left, and right shear to a maximum of 500 N. Intervertebral motions, disc pressure, and vertebral body deformations were recorded for all loads. The deformations were measured using strain gauge rosettes at three points on the inferior vertebral body and one on the superior endplate of the inferior vertebra. The disc pressures and endplate deformations measured were significantly less in shear loading compared to compression and did not change significantly with the type of compression load. Vertebral rim strains were generally greater under shear loading compared with compression. The mechanics of load transfer in compression was the production of high disc pressures which were not linearly correlated with the central endplate deformation. In shear, the mechanism appears to be via the annulus fibrosus without the development of significant disc pressure. These differences between compression and shear loading may have implications for injury mechanisms in the thoracolumbar spine.

The differential in vitro and in vivo responses of bone marrow stromal cells on novel porous gelatin-alginate scaffolds.

Tissue engineering and stem cell therapy hold great potential of being able to fully restore, repair and replace damaged, diseased or lost tissues in the body. Biocompatible porous scaffolds are used for the delivery of cells to the regeneration sites. Marrow stromal cells (MSCs), also referred to as mesenchymal stem cells, are an attractive cell source for tissue engineering, due to the relative ease of isolation and the ability of in vitro expanded MSCs to generate multiple cell types, including osteoblasts, chondrocytes and adipocytes. This study utilized a novel technique called microwave vacuum drying to fabricate porous gelatin–alginate scaffolds for the delivery of MSCs and investigated the differential in vitro and in vivo responses of MSCs seeded on these scaffolds. Scaffold total porosity was found to decrease with increased cross‐link density but the pore size and pore size distribution were not affected. Although highly porous, the scaffold had relatively small pores and limited interconnectivity. The porous gelatin–alginate scaffold demonstrated excellent biocompatibility with neovascularization on the surfaces and was bioresorbed completely in vivo, depending upon the cross‐link density. MSCs were able to attach and proliferate at the same rate on the scaffolds, and the self‐renewal potential of MSC cultures was similar during both in vitro culture and in vivo implantation. However, the subcutaneous microenvironment was found to suppress MSC differentiation along the osteogenic, chondrogenic and adipogenic lineages compared to in vitro conditions, highlighting the differential responses of MSCs cultured in vitro compared to implantation in vivo. Copyright

Allograft impaction and cement penetration after revision hip replacement: A HISTOMORPHOMETRIC ANALYSIS IN THE CADAVER FEMUR

We studied various aspects of graft impaction and penetration of cement in an experimental model. Cancellous bone was removed proximally and local diaphyseal lytic defects were simulated in six human cadaver femora. After impaction grafting the specimens were sectioned and prepared for histomorphometric analysis. The porosity of the graft was lowest in Gruen zone 4 (52%) and highest in Gruen zone 1 (76%). At the levels of Gruen zones 6 and 2 the entire cross-section was almost filled with cement. Cement sometimes reached the endosteal surface in other Gruen zones. The mean peak impaction forces exerted with the impactors were negatively correlated with the porosity of the graft.

Effects of continuous and pulsatile PTH treatments on rat bone marrow stromal cells.

Bone marrow stromal cells (MSCs) differentiation and proliferation are controlled by numerous growth factors and hormones. Continuous parathyroid hormone (PTH) treatment has been shown to decrease osteoblast differentiation, whereas pulsatile PTH increases osteoblast differentiation. However, the effects of PTH treatments on MSCs have not been investigated. This study showed continuous PTH treatment in the presence of dexamethasone (DEX) promoted osteogenic differentiation of rat MSCs in vitro, as demonstrated by increased alkaline phosphatase (ALP) activity, number of ALP expressing cells, and up-regulation of PTH receptor-1, ALP, and osteocalcin mRNA expressions. In contrast, pulsatile PTH treatment was found to suppress osteogenesis of rat MSCs, possibly by promoting the maintenance of undifferentiated cells. Additionally, the observed effects of PTH were strongly dependent on the presence of DEX. MSC proliferation however was not influenced by PTH independent of treatment regimen and presence or absence of DEX. Furthermore, our work raised the possibility that PTH treatment may modulate stem/progenitor cell activity within MSC cultures.

Microtopographical regulation of adult bone marrow progenitor cells chondrogenic and osteogenic gene and protein expressions.

Microtopographic features affect diverse cell behaviors. Adult bone marrow progenitor cells (AMPCs) constitute a multipotent heterogeneous population. We hypothesized that microtopographies could direct AMPCs lineage-specific differentiation. AMPCs isolated from Sprague-Dawley rats were CD45 depleted, expanded, and plated at 10(5) cells/cm2 on epoxy-microfabricated: (1) 60-microm-deep grooves with 95-microm pitch (D60P95), (2) 55-microm-wide and 10-microm-deep squares (W55D10), (3) 30-microm-deep grooves with 45-microm pitch (D30P45), (4) 17-microm-wide and 10-microm-deep pillars (W17D10), and (5) smooth control. AMPCs were cultured using expansion, chondrogenesis, or osteogenesis supporting media. Cell cultures were examined by scanning electron microscopy, qRT-PCR, and immunostaining at 2, 9, 16, and 23 days after plating. Expressions of osteogenesis-related genes, such as Runx-2, alkaline phosphatase, osteopontin, osteocalcin, and parathyroid hormone-related protein receptor (PTHr), and chondrogenesis-associated genes, such as Sox-9, type II collagen, and aggrecan, were determined. In expansion medium, W55D10 induced a transient increase of Sox9 expression. Compared with smooth surfaces, type II collagen mRNA and protein expressions in chondrogenic medium were significantly upregulated on W55D10 by day 23. In contrast, osteocalcin and PTHr expressions were significantly increased on D30P45 in osteogenic medium. We have demonstrated that W55D10 and D30P45 enhanced AMPCs chondrogenic and osteogenic terminal differentiation with appropriate culture conditions.

Stress distribution and consolidation in cartilage constituents is influenced by cyclic loading and osteoarthritic degeneration

The understanding of load support mechanisms in cartilage has evolved with computational models that better mimic the tissue ultrastructure. Fibril-reinforced poroelastic models can reproduce cartilage behaviour in a variety of test conditions and can be used to model tissue anisotropy as well as assess stress and pressure partitioning to the tissue constituents. The goal of this study was to examine the stress distribution in the fibrillar and non-fibrillar solid phase and pressure in the fluid phase of cartilage in axisymmetric models of a healthy and osteoarthritic hip joint. Material properties, based on values from the literature, were assigned to the fibrillar and poroelastic components of cartilage and cancellous and subchondral compact bone regions. A cyclic load representing walking was applied for 25 cycles. Contact stresses in the fibrillar and non-fibrillar solid phase supported less than 1% of the contact force and increased only minimally with load cycles. Simulated proteoglycan depletion increased stresses in the radial and tangential collagen fibrils, whereas fibrillation of the tangential fibrils resulted in increased compressive stress in the non-fibrillar component and tensile stress in the radial fibrils. However neither had an effect on fluid pressure. Subchondral sclerosis was found to have the largest effect, resulting in increased fluid pressure, non-fibrillar compressive stress, tangential fibril stress and greater cartilage consolidation. Subchondral bone stiffening may play an important role in the degenerative cascade and may adversely affect tissue repair and regeneration treatments.

Effect of bone graft substitute on marrow stromal cell proliferation and differentiation

Marrow stromal cells (MSCs) are ideally suited for tissue engineered bone grafts since they have the potential to regenerate bone, but may also maintain the homeostasis of the repaired tissue through their ability for self-renewal. An ideal bone graft substitute should support MSC self-renewal as well as differentiation to ensure complete bone defect regeneration and maintenance. The purpose of this investigation was to determine the effect of different substrate materials on MSC expansion and differentiation. Calcium polyphosphate (CPP), bone and hydroxyapatite/tricalcium phosphate (HA/TCP) were seeded with rat MSCs and maintained in culture conditions that promote cell expansion. At 0, 3, 7, 14, and 21 days cell numbers were determined by measuring their metabolic activity using a MTT assay and the frequency of cycling cells by 24 hr BrdU incorporation. Osteogenic, chondrogenic, and adipogenic marker expression in these cultures was measured by qRT-PCR. An initial drop in cell numbers was observed on all substrates. CPP and bone, but not HA/TCP supported an increase in proliferating cells at day 14 and 21. In addition, no upregulation of mature bone markers was observed in cells cultured on CPP and bone, which suggests that these substrates support the expansion of undifferentiated MSCs. In contrast, cell numbers on HA/TCP decreased with time and only rare BrdU positive cells were observed. This decrease in proliferation correlated with the down regulation of osteogenic progenitor markers and the substantial increase in mature osteocyte markers, indicating that HA/TCP favors MSC differentiation and maturation along the osteogenic lineage.

The acetabular labrum: a review of its function

UNLABELLED The acetabular labrum is a soft-tissue structure which lines the acetabular rim of the hip joint. Its role in hip joint biomechanics and joint health has been of particular interest over the past decade. In normal hip joint biomechanics, the labrum is crucial in retaining a layer of pressurised intra-articular fluid for joint lubrication and load support/distribution. Its seal around the femoral head is further regarded as a contributing to hip stability through its suction effect. The labrum itself is also important in increasing contact area thereby reducing contact stress. Given the labrums role in normal hip joint biomechanics, surgical techniques for managing labral damage are continuously evolving as our understanding of its anatomy and function continue to progress. The current paper aims to review the anatomy and biomechanical function of the labrum and how they are affected by differing surgical techniques. TAKE HOME MESSAGE The acetabular labrum plays a critical role in hip function and maintaining and restoring its function during surgical intervention remain an essential goal. Cite this article: Bone Joint J 2016;98-B:730-5.

In vivo evaluation of calcium polyphosphate for bone regeneration

Current problems associated with bone allografts include risk of disease transmission, limited availability, and cost. Synthetic scaffolds have been proposed as substitute graft materials to address these issues. Calcium polyphosphate is a novel synthetic scaffold material that has shown good mechanical properties and biocompatibility. Here, we evaluated calcium polyphosphate in terms of its ability to support cell proliferation and differentiation in vivo. Calcium polyphosphate, morsellized cancellous bone, and hydroxyapatite/tricalcium phosphate particles were seeded with marrow stromal cells and implanted subcutaneously in the back of NOD/Scid mice. At 7, 14, and 28 days the samples were harvested and the proliferation characteristics and gene expression were analyzed. All tested graft materials had similar proliferation characteristics and gene expression. The subcutaneous environment had a stronger impact on the proliferation and differentiation of the cells than the scaffold material itself. However, it was shown that calcium polyphosphate is superior to hydroxyapatite/tricalcium phosphate and bone in its ability to support cell survival in vivo. The study confirmed that calcium polyphosphate has potential for replacing morsellized cancellous bone as a graft material for bone regeneration.