Adam Briscoe

A comparison of polymer and polymer–hydroxyapatite composite tissue engineered scaffolds for use in bone regeneration. An in vitro and in vivo study

Previous in vitro work demonstrated porous PLA and PLGA both had the mechanical strength and sustained the excellent skeletal stem cell (SSC) growth required of an osteogenic bonegraft substitute, for use in impaction bone grafting. The purpose of this investigation was to assess the effects of the addition of hydroxyapatite (HA) to the scaffolds before clinical translation. PLA, PLA+10% HA, PLGA, and PLGA+10% HA were milled and impacted into discs before undergoing a standardized shear test. Cellular compatibility analysis followed 14 days incubation with human skeletal stems cells (SSC). The best two performing polymers were taken forward for in vivo analysis. SSC seeded polymer discs were implanted subcutaneously in mice. All polymers had superior mechanical shear strength compared with allograft (p < 0.01). Excellent SSC survival was demonstrated on all polymers, but the PLA polymers showed enhanced osteoblastic activity (ALP assay p < 0.01) and collagen-1 formation. In vivo analysis was performed on PLA and PLA+10% HA. MicroCT analysis revealed increased bone formation on the PLA HA (p < 0.01), and excellent neo-vessel formation in both samples. Histology confirmed evidence of de novo bone formation. PLA HA showed both enhanced osteoinductive and osteogenic capacity. This polymer composite has been selected for scaled-up experimentation before clinical translation.

Supercritical CO2 fluid-foaming of polymers to increase porosity: a method to improve the mechanical and biocompatibility characteristics for use as a potential alternative to allografts in impaction bone grafting?

Disease transmission, availability and cost of allografts have resulted in significant efforts to find an alternative for use in impaction bone grafting (IBG). Recent studies identified two polymers with both structural strength and biocompatibility characteristics as potential replacements. The aim of this study was to assess whether increasing the polymer porosity further enhanced the mechanical and cellular compatibility characteristics for use as an osteogenic biomaterial alternative to allografts in IBG. Solid and porous poly(DL-lactide) (P(DL)LA) and poly(DL-lactide-co-glycolide) (P(DL)LGA) scaffolds were produced via melt processing and supercritical CO(2) foaming, and the differences characterized using scanning electron microscopy (SEM). Mechanical testing included milling and impaction, with comparisons made using a shear testing rig as well as a novel agitation test for cohesion. Cellular compatibility tests for cell number, viability, and osteogenic differentiation using WST-1 assays, fluorostaining, and ALP assays were determined following 14 day culture with skeletal stem cells. SEM showed excellent porosity throughout both of the supercritical-foam-produced polymer scaffolds, with pores between 50 and 200 μm. Shear testing showed that the porous polymers exceeded the shear strength of allograft controls (P<0.001). Agitation testing showed greater cohesion between the particles of the porous polymers (P<0.05). Cellular studies showed increased cell number, viability, and osteogenic differentiation on the porous polymers compared to solid block polymers (P<0.05). The use of supercritical CO(2) to generate porous polymeric biodegradable scaffolds significantly improves the cellular compatibility and cohesion observed compared to non-porous counterparts, without substantial loss of mechanical shear strength. These improved characteristics are critical for clinical translation as a potential osteogenic composite for use in IBG.

From bench to clinic and back: skeletal stem cells and impaction bone grafting for regeneration of bone defects

Tissue engineering offers enormous potential for bone regeneration. Despite extensive in vitro and in vivo work, few strategies translate into clinical practice. This paper describes the combination of skeletal stem cells (SSCs) and impaction bone grafting (IBG) for the treatment of patients with bone defects associated with avascular necrosis of the femoral head. SSCs and milled allograft were impacted into necrotic bone in the femoral heads of four patients. Three patients remained asymptomatic at 22–44 month follow‐up, but one patient has required total hip replacement (both hips). This has allowed retrieval of the femoral heads, which were analysed structurally and functionally by μCT, histology and mechanical testing. A central channel of impacted bone was found in the femoral heads, which displayed a mature trabecular micro‐architecture. The impacted bone was denser than the surrounding trabecular bone, as strong in compression and with histological micro‐architecture comparable to that of trabecular bone. Analysis of the retrieved femoral head samples has demonstrated that this tissue‐engineering strategy regenerates bone that is both structurally and functionally analogous to normal trabecular bone. SSCs, together with IBG, have proved an effective treatment for avascular necrosis of the femoral head and offer significant potential for the broader spectrum of bone defects. Copyright

A tissue engineering strategy for the treatment of avascular necrosis of the femoral head

Background & purpose Skeletal stem cells (SSCs) and impaction bone grafting (IBG) can be combined to produce a mechanically stable living bone composite. This novel strategy has been translated to the treatment of avascular necrosis of the femoral head. Surgical technique, clinical follow-up and retrieval analysis data of this translational case series is presented. Methods SSCs and milled allograft were impacted into necrotic bone in five femoral heads of four patients. Cell viability was confirmed by parallel in vitro culture of the cell-graft constructs. Patient follow-up was by serial clinical and radiological examination. Tissue engineered bone was retrieved from two retrieved femoral heads and was analysed by histology, microcomputed tomography (μCT) and mechanical testing. Results Three patients remain asymptomatic at 22- to 44-month follow-up. One patient (both hips) required total hip replacement due to widespread residual necrosis. Retrieved tissue engineered bone demonstrated a mature trabecular micro-architecture histologically and on μCT. Bone density and axial compression strength were comparable to trabecular bone. Conclusions Clinical follow-up shows this to be an effective new treatment for focal early stage avascular necrosis of the femoral head. Unique retrieval analysis of clinically translated tissue engineered bone has demonstrated regeneration of tissue that is both structurally and functionally analogous to normal trabecular bone.

Taking tissue engineering principles into theatre: retrieval analysis from a clinically translated case

AIM Tissue engineering has enormous potential for the regeneration of bone defects. Approximately 4 years ago we reported on a 62 year old patient who underwent treatment of a benign cyst in the proximal femur by impaction bone grafting supplemented with autologous bone marrow. The cyst and symptoms subsequently recurred and this patient has now required a total hip replacement. This has provided a rare opportunity for ex vivo analysis of clinically applied tissue engineered bone. MATERIALS & METHODS The femoral head was retrieved at surgery and the structural and functional characteristics of the tissue engineered bone were analyzed by micro-computed tomography, histology and mechanical testing. RESULTS The impacted bone demonstrated a trabecular structure that contained islands of nonincorporated graft. The graft was denser than the patients trabecular bone with comparable strength. The cyst material had penetrated along the channel of bone and an increased number of osteoclasts were observed. DISCUSSION This study has provided detailed ex vivo analysis of retrieved human tissue engineered bone and possible reasons for the observed construct failure are discussed in this article. The impacted bone displayed some evidence of remodeled trabecular structure, although the bone marrow aspirate that was initially combined with the allograft contained a relatively low concentration of osteoprogenitor cells. Cellular augmentation was insufficient to overcome the osteoclastic process associated with renewed cyst formation. Concentration or culture expansion of osteoprogenitor cells from aspirated bone marrow is recommended for biological augmentation of bone graft.

Polymerisation stress modelling in acrylic bone cement

Fatigue failure of the cement mantle has been proposed as one of the failure processes contributing to aseptic loosening of cemented joint replacements. It has also been suggested that fatigue failure is dramatically accelerated by residual stress generated during the cement polymerisation process. Previous computational models of the polymerisation process have investigated only the latter part of polymerisation by assuming both instantaneous hardening of the material (a stress locking point) and that all residual stress results from thermal shrinkage after this stress locking point. In this study, finite element models which use the local degree of polymerisation to calculate material properties and shrinkage have been used to predict residual stresses in two models of total hip replacement cement mantles. Results indicate that the final value of cement mantle stress may not be the highest stresses that the cement is subjected to during the polymerisation process. Two models are presented, a 2-dimensional model, which was adapted from a similar model in the literature (Lennon and Prendergast, 2002) and a 3-dimensional concentric-cylinders model. In both cases a chemical kinetics model was used to predict the progress of the polymerisation reaction and a second linear model used to predict cement mechanical properties and density, and so stress generation and volume change, over time. There was good agreement of the results of the 2D model with its counterpart in the literature. For the 3D model, the final residual stress magnitudes and patterns showed good agreement with similar physical and computational models in the literature.

An analysis of polymer type and chain length for use as a biological composite graft extender in impaction bone grafting: a mechanical and biocompatibility study

Impaction bone grafting (IBG) with human allograft remains the preferred approach for replacement of lost bone stock during revision hip surgery. Associated problems include cost, disease transmission, and stem subsidence. Synthetic grafts are therefore appealing, and ideally display similar mechanical characteristics as allograft, but with enhanced ability to form de novo bone. High and low molecular weight forms of three different polymers [poly(DL-lactide) (P(DL) LA), poly(DL-lactide-co-glycolide) (P(DL) LGA), and poly(ε-caprolactone) (PCL)] were milled, impacted into discs, and then examined in a shear testing rig, in comparison to allograft. In addition, skeletal stem cells (SSCs) were combined with each of the milled polymers, followed by impaction and examination for cell viability and number, via fluorostaining and biochemical assays. The shear strengths of high/low mwt P(DL) LA, and high/low mwt P(DL) LGA were significantly higher than allograft (p < 0.01). High/low mwt PCL had significantly lower shear strengths (p < 0.01). WST-1 assay and fluorstaining indicated significantly increased cell viability on high mwt P(DL) LA and high mwt P(DL) LGA over allograft (p < 0.05). Mechanical and biochemical analysis indicated improved properties of high mwt P(DL) LA and high mwt P(DL) LGA over allograft. This study indicates the potential of these polymers for use as substitute human allograft, creating a living composition with SSC for application in IBG.

The Biotribology of PEEK-on-HXLPE Bearings Is Comparable to Traditional Bearings on a Multidirectional Pin-on-disk Tester

BackgroundAll-polymer bearings involving polyetheretherketone (PEEK) have been proposed for orthopaedic applications because they may reduce stress shielding, reduce weight of the implants, reduce wear and risk of osteolysis, and prevent release of metal ions by replacing the metal articulating components. Little is known about the biotribology of all-polymer PEEK bearings, including the effects of cross-shear, which are relevant for implant longevity, especially in the hip, and increased temperature that may affect lubricant proteins and, hence, lubrication in the joint.Questions/purposesUsing pin-on-disk in vitro testing, we asked: (1) Can all-polymer bearing couples involving PEEK have a comparable or lower wear rate than highly crosslinked UHMWPE (HXLPE) on CoCr bearing couples? (2) Is the wear rate of PEEK bearing couples affected by the amount of cross-shear? (3) Is there a difference in wear mechanism and surface morphology for all-polymer bearing surfaces compared with UHMWPE (HXLPE) on CoCr?MethodsWe simultaneously tested a total of 100 pin-on-disk couples (n = 10 per bearing couple) consisting of three traditional metal-on-UHMWPE and seven polymer-on-polymer bearings for 2 million cycles under physiologically relevant conditions and in accordance with ASTM F732. Using analysis of variance, we analyzed the effect of bearing surface topography and cross-shear on wear rate. The changes in surface topography were evaluated using optical microscopy. Sample size was sufficient to provide 80% power to detect a difference of 1.4 mm3/MC in average wear rates of bearing couples.ResultsThe combined wear rates of all-polymer bearing couples were not different than traditional bearing couples. With the numbers available, the PEEK and HXLPE bearing couple had a mean wear rate (WR: mean ± SD) of 0.9 ± 1.1 mm3/MC (95% confidence interval [CI], 0.2–1.5 mm3/MC), which was not different than the wear rate of the CoCr and HXLPE bearing couple (1.6 ± 2.0 mm3/MC; 95% CI, 0.4–2.8 mm3/MC; mean difference = 0.73 mm3/MC, p = 0.36). Bearing couples with PEEK reinforced with a carbon fiber (CFR-PEEK) counterface had higher wear rates (14.5 ± 15.1 mm3/MC; 95% CI, 9.1–20.0 mm3/MC) than bearing couples with a PEEK (5.1 ± 3.7 mm3/MC; 95% CI, 3.7–6.4 mm3/MC) or CoCr (4.1 ± 2.7 mm3/MC; 95% CI, 3.2–5.1 mm3/MC) counterface (mean difference = 9.5 mm3/MC, p < 0.001; and mean difference = 10.4 mm3/MC, p < 0.001, respectively). PEEK and HXLPE were insensitive to the cross-shear scenario in the contact mechanics (WR: 0.3 ± 0.1 mm3/MC for PEEK pins [95% CI, 0.2–0.3 mm3/MC] [representing full cross-shear condition] and 0.0 ± 1.0 mm3/MC for PEEK disks [95% CI, −0.5 to 0.5 mm3/MC] [representing limited cross-shear condition], mean difference = 0.3 mm3/MC, p = 0.23; WR: 1.3 ± 1.0 mm3/MC for HXLPE pins [95% CI, 0.7–1.9 mm3/MC] [full cross-shear] and 2.1 ± 2.2 mm3/MC for HXLPE disks [95% CI, 0.8–3.3 mm3/MC] [limited cross-shear], mean difference = 0.8 mm3/MC, p = 0.24). Qualitatively, the surface morphology of UHMWPE appeared similar with PEEK or CoCr as a counterface, although it had a rougher appearance when coupled with carbon fiber-reinforced PEEK. No transfer film was detected on the specimens.ConclusionsOur in vitro pin-on-disk data suggest that all-polymer bearings, especially PEEK-on-HXLPE bearing couples, may represent a viable alternative to traditional bearings with respect to their wear performance. Our results warrant further testing of all-polymer bearing couples in physiologically relevant joint simulator tests.Clinical RelevanceThe in vitro pin-on-disk wear resistance of all-polymer bearings incorporating PEEK-on-HXLPE warrants further investigation using joint simulator testing for their validation as useful, metal-free alternatives to traditional CoCr-on-HXLPE bearings for use in orthopaedic applications.

Do the Cup Surface Properties Influence the Initial Stability

This project tests the relationship between the acetabular cup surface characteristics and their initial stability by comparing uncemented (Trabecular Metal (TM) and Trilogy) and cemented polyethylene shells. We hypothesised that different surface properties of uncemented cups will influence the cup stability. Mounted directly onto host bone, TM and cemented cups were significantly more stable than Trilogy cups (P < 0.01), with minimal difference between TM and cemented cups (P > 0.1). On 100% graft bed, there was marginal difference between all three cup types (P > 0.1). Incremental cavitary and segmental defects resulted in reducing stability, with cemented cups being minimally more stable (P > 0.1). TM cups possess satisfactory initial stability in bone graft constructs. This study demonstrates that TM shells are marginally less stable than cemented cups in the absence of significant host bone contact.

MEDIAL-LATERAL LOADING AND WEAR IN TKA

Pre-clinical wear testing of Total Knee Arthroplasty has traditionally been carried out in a physical experiment [Fisher, 2002]. Recent computational models have been shown to have sufficient accuracy to be considered alongside these experiments [Knight, 2007]. These computational models use a relationship of wear volume proportional to the product of contact pressure, sliding distance and cross shear [Maxian, 1996]. Instrumented knees have recently shown that medial-lateral (ML) loads may be of similar magnitudes to that of the anterior-posterior (AP) load. The AP load is known to have a significant effect on the kinematics of the total knee replacement and so it is reasonable to assume that application of an ML load may have a similar degree of influence on kinematics. The effect of the ML load is hypothesised to increase the cross shear and hence the wear rate. At present, the ISO standard for testing TKA contains no provision for a ML load.