Less blood loss and operative times were found with skip laminectomy. Similar degrees of decompression with both techniques. Significantly improved axial pain scores with skip laminectomy. Significantly improved preservation of range of movement with skip laminectomy.
Tissue engineering is a rapidly expanding field of research. Bone and cartilage engineering are being undertaken in an attempt to treat osteoarthritis and repair bone defects. In spite of extensive research little successful clinical application of this work has been seen. There are however many advances in the field that one day may have therapeutic interest. One particular area of interest is the potential for using osteophyte tissue in repairing osteoarthritic defects. Osteophytes represent an attempt by the body to regenerate bone and cartilage. They present an obvious source of cells for tissue engineering. Research ay QUT has shown that cells within the osteophytes are a better source of bone and cartilage regeneration in the laboratory than matched patient’s bone marrow stem cells. Osteoarthritis remains the ultimate challenge for orthopaedic tissue engineering. Understanding the chemical and mechanical signals occurring in osteoarthritis presents opportunities for targeted drug delivery and potential slowing of disease. We have identified changes within the MMP profile of cells at the osteochondral junction. Subchondral sclerosis appears to be associated with changes in the nature of chondrocytes deep within the cartilage layer. This transformation of chondrocytes into osteoblast-like tissue in many ways mimics the changes seen in the growth plate once maturity is reached. Understanding the parallels between these processes may help answer some of the mechanisms of the development of osteoarthritis. This talk will discuss the above topics as well as other areas of interest to an orthopaedic surgeon working within a group of 10 cell biologists.
This presentation introduces a new tool to be used in the cementing of acetabular components in total hip arthroplasty, the ‘Rim Cutter’. The Rim Cutter is designed to cut a ledge in the rim of the acetabulum into which a flanged cup can be cemented. The flange is trimmed such that it fits precisely into the ledge cut in the acetabulum. We present the in vitro pilot study of the effect of using this tool on the intra-acetabular cement mantle pressure during cup insertion and also the effect on the depth of cement penetration as the cup is inserted. A significant improvement in both cement pressure and cement penetration over conventional flanged and unflanged cups is noted. Improved cement penetration around the rim of the acetabulum in THR has implications for reducing the rate of aseptic loosening. The pilot study also suggests other beneficial features of using the rim cutter such as improved cup centralisation, control of orientation and the prevention of the cup ‘bottoming out’. Further in vivo studies are required to better assess its efficacy.
To regenerate the complex tissue such as bone-cartilage construct using tissue engineering approaches, controllable differentiation of mesenchymal stem cells (BMSCs) into chondrogenic and osteogenic lineages is crucially important. Although bilayered scaffolds have been investigated in vitro and in vivo, no culture system is available to test BMSCs differentiation into bone and cartilage simultaneously in bilayered scaffolds. This study investigated a defined culture media, which supported osteoblast and chondrocyte differentiation depending on growth factors implemented in biomaterials. In 2-dimensional culture, BMSCs differentiated to chondrocytes when transforming growth factor-beta 3 (TGF-β3) was added to the defined media, whereas osteogenic differentiation was induced by adding bone morphogenetic protein 7 (BMP-7). BMSC differentiation to osteogenic and chondrogenic lineages was further strengthen in 3-dimensional culture. Proteoglycan formation, type II collagen, and aggrecan were upregulated in the defined media when BMSCs were mixed with fibrin gel impregnated with TGF-β3. Mineralization and the expression of osteogenic markers such as alkaline phosphatase, osteopontin, and osteoclacin were noticeable when BMSCs cultured in hydroxyapatite-tricalcium phosphate (HA/TCP) scaffolds coated with BMP-7. This study generated and tested a growth media, which could induce osteogenic and chondrogenic differentiation of BMSCs in one culture system. These results will help the development of tissue substitutes for multi-complexed tissues such as subchondral replacement.
Bone Tissue Engineering Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia Adult mesenchymal stem cells (MSCs) are a topic of immense research interest in the field of tissue engineering. Since, depletion of multipotent cells has been implicated in degenerative joint diseases, cell based therapies have been proposed for tissue regeneration, especially for cartilage repair. The aim of the present study is to focus on the possibility of deriving and expanding multipotential MSCs from the heterogeneous bone marrow stromal samples of patients with osteoarthritis (OA) by characterising MSCs at the single cell level. Single cell clonal cultures were established by limiting dilution of marrow stromal cells from three OA patients. A total of 14 clones with a wide variation in their cell doubling time were isolated. The clones were grouped into fast-growing and slow-growing clones. All except one of the fast-growing clones were tripotential. However the slow-growing clones showed limited differentiation potential and morphological changes associated with cellular senescence with extended duration in culture. Flow cytometric analysis did not depict any difference in the expression of the selected putative MSC cell surface markers CD29, CD44, CD90, CD105 and CD166 between fast-growing and slow-growing clones indicating a strong need to investigate for novel cell-surface markers. Further, proteomic analysis to understand the sub-cellular processes responsible for the existence of varying sub-populations identified 11 differentially expressed proteins. These proteins were reported to be associated with cellular organization, signal transduction, energy pathways and stress related proteins. Identification of signaling pathway proteins and cell cycle related proteins, such as calmodulin and caldesmon in the clonal populations, suggest that high-throughput proteomic technologies like two dimensional liquid chromatography (2D LC) coupled with mass spectrometry (MS) may facilitate the discovery of therapeutically useful biomarkers. This study demonstrated the existence of a fast-growing multipotential MSC population from bone marrow samples of patients with OA. Therefore, despite a supposedly smaller stem cell compartment in these patients, we demonstrate here that they can still yield a potentially therapeutically useful source of syngeneic MSCs.
Synthetic biodegradable polymers have been utilized increasingly in pharmaceutical, medical and biomedical engineering. Control of the interaction of living cells and biomaterials surfaces is one of the major goals in the design and development of new polymeric biomaterials in tissue engineering. In this study, a novel amphiphilic tri-block copolymer, methoxy-terminated poly (ethylene glycol) (MPEG) – polyL-lactide (PLLA) – polylysine (PLL) was synthesized. Various molecular compositions of tri-block copolymers were prepared via optimising the parameters and characterized through Nuclear Magnetic Resonance and Gel Permeation Chromatography. The tri-block copolymer was then mixed with high molecular weight PLLA to form a flat film. The surface properties measured by X-ray Photoelectron Spectroscopy and Atomic Force Microscopy demonstrated high content of the PLL on the surface of PLLA film, which indicated self-segregation of MPEG-b-PLLA-b-PLL on PLLA surface. No cytotoxicity was detected in triblock copolymers, and compared to pure PLLA and diblock copolymers, the triblock copolymers were much more effective for cell adhesion and proliferation. It was noted that the hydrophilic chain of PEG and PLL stretched out and formed an outer layer, especially under the aqueous environment, which resulted in enhanced cell attachment and proliferation. The self-segregation behaviour of MPEG-b-PLLA-b-PLL triblock copolymer shows a potential application in scaffold preparation of tissue engineering.
Interactions between cells and polymers are mediated by proteins, which are either secreted by cells and immobilized on the biomaterial surface, or absorbed from the medium. Poly (lactic acid) (PLA) is widely used in tissue engineering as a scaffold material, however anchorage-dependent cells such as osteoblasts do not attach, grow, and differentiate well on a hydrophobic surface. In this study, a hydrophilic polymer-poly (ethylene glycol) (PEG) was used to develop diblock polymers, Methoxy-terminated poly (ethylene glycol)-Poly (lactic acid) (MPEG-PLA) to investigate cell-biomaterial interactions. Osteoblasts were cultured on different composition of PEG-PLA films in serum free or serum condition. Lactate dehydrogense (LDH) assay was used to assess the cytotoxicity of the copolymers and cell attachment and proliferation on the polymer surfaces; furthermore cell morphology was visualized by Crystal Violet stain. The results showed that MPEG-PLA films induced early osteoblast attachment in serum free condition and the higher content of PEG in the MPEG-PLA films the more cell attachment was noticed. No significant difference of cell attachment was observed on MPEG-PLA films between serum free and 10% serum culture condition. Crystal Violet stain demonstrated the same trend in the cell-spreading characteristics on the polymer surface. In conclusion MPEG-PLA copolymer can enhance osteoblast attachment under serum-free condition, which implies a potential application in cell delivery therapy due to the restriction in animal products for human therapeutically goods.
In both physiological and pathological processes, periosteum plays a determinant role in both bone formation and fracture healing. However, no specific reports are available so far focusing on the detailed structural and major cellular differences between the periostea covering different bone surface areas in relation to ageing. The aim of this study is to compare the structural and cellular differences in diaphyseal and epiphyseal periostea in different-aged rats using histological and immunohistochemical methods. Four female Lewis rats from each group of juvenile (7-week old), mature (7-month old) and aged groups (2-year old) were sacrificed and the right femur of each rat was retrieved, fixed, decalcified and embedded. 5μm thick serial sagittal sections were cut and stained with Hematoxylin and Eosin, Stro-1 (stem cell marker), F4/80 (macrophage marker), TRAP (osteoclast marker) and vWF (endothelial cell marker). 1mm length of middle diaphyseal and epiphyseal periosteum were selected for observation. The thickness, total cell number and positive cell number for each antibody in each periosteal area and different-aged groups were measured and compared. The results were subjected to ANOVA and SNK-q tests. The results showed that the thickness and cell number in diaphyseal periosteum decreased with age (p<
0.001). In comparison with diaphyseal area, the thickness and cell number in epiphyseal periosteum were much higher (p<
0.001). There were no significant differences between the juvenile and aged groups in the thickness and cell number in cambial layer of epiphyseal periosteum (p>
0.05). However, the juvenile rats had more Stro1+, F4/80+ cells and blood vessels and few TRAP+ cells in different periosteal areas compared with other groups(p<
0.001). The aged rats showed much less Stro1+ cells, but more F4/80+,TRAP+ cells and blood vessels in the cambial layer of epiphyseal periosteum (p<
0.001). In conclusion, the age-related structure and cell population in diaphyseal and epiphyseal periostea are different, especially in aged rats. The epiphyseal periosteum of aged rats seems more destructive than diaphyseal part and other age groups. Macrophages in the periosteum play a dual important role in osteogenesis and osteoclastogenesis.
Bone Tissue Engineering Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, QLD 4059, Australia. Osteophytes are the most remarkable and consistently distinct feature of osteoarthritis (OA). Their formation may be related to pluripotential cells in the periosteum responding to stimulus during OA. This study aimed to isolate stem cells from osteophyte tissues, and characterise their phenotype, proliferation and differentiation potential, and immuno-modulatory properties. Osteophyte derived cells were isolated from five osteophyte tissue samples collected during knee replacement surgery. These cells were characterised by the expression of cell surface antigens, differentiation potential into mesenchymal lineages, growth kinetics and modulation of allo-immune responses. Multipotential stem cells (MSCs) were identified from all osteophyte samples namely osteophyte derived MSCs (oMSCs). The surface antigen expression of oMSCs was consistent with that of mesenchymal stem cells, such as lacking the haematopoietic and common leukocyte markers (CD34, CD45) while expressing those related to adhesion (CD29, CD166, CD44) and stem cells (CD90, CD105, CD73). The longevity of oMSCs in culture was superior to that of bone marrow derived MSC (bMSCs), and they readily differentiated into tissues of the mesenchymal lineages. oMSCs also demonstrated the ability to suppress allogeneic T-cell proliferation, which was associated with the expression of tryptophan degrading enzyme indoleamine 2,3 dioxygenase (IDO). Our results showed that osteophyte derived cells had similar properties to mesenchymal stem cells in the expression of antigen phenotype, differential potential and suppression of allo-immune response. Furthermore, when compared to bMSCs, oMSCs maintained a higher proliferative capacity, which may offer an alternative source for therapeutic stem cell based tissue regeneration.
We are comparing the above two techniques with 25 patients in each group operated by 3 surgeons. The first group had conventional laminoplasty and the second group underwent the skip laminectomy. The groups were comparable in age, sex, pathology and clinical presentation. Both these group had clinical outcome measurements using SF 12 questionnaires, pre and postoperative clinical assessment with standard tools performed by independent surgeon and a specialist spinal physiotherapist. We also routinely performed pre and postoperative MRI scans to assess the adequacy of decompression.
58,109 primary hip replacements for osteoarthritis were investigated for effect of age group, sex and fixation method. Age group and sex were not significant risk factors in revision for dislocation. Studying fixation method, cementless acetabular components were implanted more frequently (49,027, 84%) than cemented (9,082, 15.6%). In total, there were 428 (0.7%) revisions for dislocation, 369(0.8%) with a cementless acetabulum and 59 (0.6%) with cemented. Relative risk (cementless v cemented acetabulum adjusted for age group, sex and head size) of 1.59 (CI 1.19 to 2.12, p<
0.01). Head sizes of >
30mm, 28mm, 26mm and 22mm had significantly increasing relative risk (p<
0.001).
There was no significant correlation between clinical leg length change, measured in the operating theatre and the leg length change predicted by navigation. Accuracy of cup and stem placement was assessed by comparison of the homogeneity of variances, the Levene statistic, in the navigated and control groups. The range of cup inclination, cup version and stem version was significantly narrowed in the navigation group (p<
0.05).
We are comparing the above two techniques with 25 patients in each group operated by 3 surgeons. The first group had conventional laminoplasty and the second group underwent the skip laminectomy. The groups were comparable in age, sex, pathology and clinical presentation. Both these group had clinical outcome measurements using SF 12 questionnaires, pre and postoperative clinical assessment with standard tools performed by independent surgeon and a specialist spinal physiotherapist. We also routinely performed pre and postoperative MRI scans to assess the adequacy of decompression.
The compressive strength of the MAA and MSA based copolymers was measured as a function of anhydride concentration. Compressive strength for MMA increased (90±9 to 140±10 Mpa) in an approximately linear manner for MAA concentrations from 10 to 40 wt.% but decreased markedly for MAA concentration of 45% (62±14 Mpa). The compressive strength of MSA decreased exponentially for concentrations ranging from 10 to 45 wt.% (140±18 to 39±1 Mpa).
Computer aided joint replacement surgery has become very popular during recent years and is being done in increasing numbers all over the world. The accuracy of the system depends to a major extent, on accurate registration and immobility of the tracker attachment devices to the bone. This study was designed to assess the forces needed to displace the tracker attachment devices in the bone simulators. Bone simulators were used to maintain the uniformity of the bone structure during the study. The fixation devices tested were 3mm diameter self drilling, self tapping threaded pin, 4mm diameter self tapping cortical threaded pin, 5mm diameter self tapping cancellous threaded pin and a triplanar fixation device ‘ortholock’ used with three 3mm pins. All the devices were tested for pull out, translational and rotational forces in unicortical and bicortical fixation modes. Also tested was the normal bang strength and forces generated by leaning on the devices. The forces required to produce translation increased with the increasing diameter of the pins. These were 105 N, 185 N, and 225 N for the unicortical fixations and 130N, 200N, 225 N for the bicortical fixations for 3mm, 4mm and 5 mm diameter pins respectively. The forces required to pull out the pins were 1475N, 1650N, 2050N for the unicortical, 1020N, 3044N and 3042N for the bicortical fixated 3mm, 4mm and 5mm diameter pins. The ortholock translational and pull out strength was tested to 900N and 920N respectively and still it did not fail. Rotatory forces required to displace the tracker on pins was to the magnitude of 30N before failure. The ortholock device had rotational forces applied up to 135N and still did not fail. The manual leaning forces and the sudden bang forces generated were of the magnitude of 210 N and 150 N respectively. The strength of the fixation pins increases with increasing diameter from three to five mm for the translational forces. There is no significant difference in pull out forces of four mm and five mm diameter pins though it is more than the three mm diameter pins. This is because of the failure of material at that stage rather than the fixation device. The rotatory forces required to displace the tracker are very small and much less than that can be produced by the accidental leaning or bang produced by the surgeon or assistants in single pins. Although the ortholock device was tested to 135 N in rotation without failing, one has to be very careful not to put any forces during the operation on the tracker devices to ensure the accuracy of the procedure.