Fortifying the Bone-Implant Interface Part 1: An In Vitro Evaluation of 3D-Printed and TPS Porous Surfaces.

BACKGROUND: An aging society and concomitant rise in the incidence of impaired bone health have led to the need for advanced osteoconductive spinal implant surfaces that promote greater biological fixation (e.g. for interbody fusion cages, sacroiliac joint fusion implants, and artificial disc replacements). Additive manufacturing, i.e. 3D-printing, may improve bone integration by generating biomimetic spinal implant surfaces that mimic bone morphology. Such surfaces may foster an enhanced cellular response compared to traditional implant surfacing processes. METHODS: This study investigated the response of human osteoblasts to additive manufactured (AM) trabecular-like titanium implant surfaces compared to traditionally machined base material with titanium plasma spray (TPS) coated surfaces, with and without a nanocrystalline hydroxyapatite (HA) coating. For TPS-coated discs, wrought Ti6Al4V ELI was machined and TPS-coating was applied. For AM discs, Ti6Al4V ELI powder was 3D-printed to form a solid base and trabecular-like porous surface. The HA-coating was applied via a precipitation dip-spin method. Surface porosity, pore size, thickness, and hydrophilicity were characterized. Initial cell attachment, proliferation, alkaline phosphatase (ALP) activity, and calcium production of hFOB cells (n=5 per group) were measured. RESULTS: Cells on AM discs exhibited expedited proliferative activity. While there were no differences in mean ALP expression and calcium production between TPS and AM discs, calcium production on the AM discs trended 48% higher than on TPS discs (p=0.07). Overall, HA-coating did not further enhance results compared to uncoated TPS and AM discs. CONCLUSIONS: Results demonstrate that additive manufacturing allows for controlled trabecular-like surfaces that promote earlier cell proliferation and trends toward higher calcium production than TPS coating. Results further showed that nanocrystalline HA may not provide an advantage on porous titanium surfaces. CLINICAL RELEVANCE: Additive manufactured porous titanium surfaces may induce a more osteogenic environment compared to traditional TPS, and thus present as an attractive alternative to TPS-coating for orthopedic spinal implants.

Fortifying the Bone-Implant Interface Part 2: An In Vivo Evaluation of 3D-Printed and TPS-Coated Triangular Implants.

BACKGROUND: Minimally invasive surgical fusion of the sacroiliac (SI) joint using machined solid triangular titanium plasma spray (TPS) coated implants has demonstrated positive clinical outcomes in SI joint pain patients. Additive manufactured (AM), i.e. 3D-printed, fenestrated triangular titanium implants with porous surfaces and bioactive agents, such as nanocrystalline hydroxyapatite (HA) or autograft, may further optimize bony fixation and subsequent biomechanical stability. METHODS: A bilateral ovine distal femoral defect model was used to evaluate the cancellous bone-implant interfaces of TPS-coated and AM implants. Four implant groups (n=6/group/time-point) were included: 1)TPS-coated, 2)AM, 3)AM+HA, and 4)AM+Autograft. The bone-implant interfaces of 6- and 12-week specimens were investigated via radiographic, biomechanical, and histomorphometric methods. RESULTS: Imaging showed peri-implant bone formation around all implants. Push-out testing demonstrated forces greater than 2500 N, with no significant differences among groups. While TPS implants failed primarily at the bone-implant interface, AM groups failed within bone ~2-3mm away from implant surfaces. All implants exhibited bone ongrowth, with no significant differences among groups. AM implants had significantly more bone ingrowth into their porous surfaces than TPS-coated implants (p<0.0001). Of the three AM groups, AM+Auto implants had the greatest bone ingrowth into the porous surface and through their core (p<0.002). CONCLUSIONS: Both TPS and AM implants exhibited substantial bone ongrowth and ingrowth, with additional bone through growth into the AM implants' core. Overall, AM implants experienced significantly more bone infiltration compared to TPS implants. While HA-coating did not further enhance results, the addition of autograft fostered greater osteointegration for AM implants. CLINICAL RELEVANCE: Additive manufactured implants with a porous surface provide a highly interconnected porous surface that has comparatively greater surface area for bony integration. Results suggest this may prove advantageous toward promoting enhanced biomechanical stability compared to TPS-coated implants for SI joint fusion procedures.

The effects of multiple high-resolution peripheral quantitative computed tomography scans on bone healing in a rabbit radial bone defect model.

The use of in vivo high-resolution computed tomography (CT) scanners provides the unique opportunity for evaluating temporal progression in healing of bone defects. However, these in vivo scanners impose ionizing radiation that could affect the healing and morphology of the bone. The primary objective of this study was to determine the effects of in vivo scanning at 2-week intervals on bone healing of a critical sized radial defect in rabbits and to investigate the effect of this radiation protocol on bone marrow cell viability using clinically applicable radiation doses. Thirty male rabbits were randomized into three groups: two groups received a 15 mm defect in the left radius that was filled with an autologous bone graft (DEF-CT and DEF-SHAM), and one group acted as an intact control (INT-CT). The duration of the study was 6 weeks. DEF-CT and INT-CT had high-resolution CT scans performed at 2-week intervals. The total cumulative radiation dose was 81.6 mGy per animal. DEF-SHAM received sham CT scans at the same time points. In group DEF-CT, the bone volume (BV) in the defect increased significantly over time (p≤0.002, for all comparisons); the bone mineral density (BMD) in the defect decreased over time and was significantly lower at weeks 4 and 6 than at weeks 0 and 2 (p<0.001, for all comparisons). In group INT-CT, BV and BMD did not change over time (p=1, for all comparison). The BV (p=0.50) and the BMD (p=0.37) in the defect as measured by microCT scan during ex vivo analysis was not significantly different between DEF-CT and DEF-SHAM. Similarly, histomorphometry showed no significant difference in the total bone area (p=0.22) and percentage bone within the defect (p=0.24) between these groups. Bone marrow analysis of the left (radiated) and right (non-radiated) radius of the INT-CT group via a Colony Forming Units (CFU) assay demonstrated an average of 25.3 and 28.5 colonies for radiated and non-radiated radii, respectively (p=0.72). In conclusion, there was no significant difference in bone healing between radiated and non-radiated radius defects in rabbits. This is an important finding as it demonstrates that serial in vivo high resolution-CT imaging can not only provide accurate tissue regeneration data, but it can also be used to reduce the number of temporal cohorts within an experimental design.

Duplication and retention biases of essential and non-essential genes revealed by systematic knockdown analyses.

When a duplicate gene has no apparent loss-of-function phenotype, it is commonly considered that the phenotype has been masked as a result of functional redundancy with the remaining paralog. This is supported by indirect evidence showing that multi-copy genes show loss-of-function phenotypes less often than single-copy genes and by direct tests of phenotype masking using select gene sets. Here we take a systematic genome-wide RNA interference approach to assess phenotype masking in paralog pairs in the Caenorhabditis elegans genome. Remarkably, in contrast to expectations, we find that phenotype masking makes only a minor contribution to the low knockdown phenotype rate for duplicate genes. Instead, we find that non-essential genes are highly over-represented among duplicates, leading to a low observed loss-of-function phenotype rate. We further find that duplicate pairs derived from essential and non-essential genes have contrasting evolutionary dynamics: whereas non-essential genes are both more often successfully duplicated (fixed) and lost, essential genes are less often duplicated but upon successful duplication are maintained over longer periods. We expect the fundamental evolutionary duplication dynamics presented here to be broadly applicable.

Centrosome duplication proceeds during mimosine-induced G1 cell cycle arrest.

Centrosome duplication must remain coordinated with cell cycle progression to ensure the formation of a strictly bipolar mitotic spindle, but the mechanisms that regulate this coordination are poorly understood. Previous work has shown that prolonged S-phase is permissive for centrosome duplication, but prolonging either G2 or M-phase cannot support duplication. To examine whether G1 is permissive for centrosome duplication, we release serum-starved G0 cells into mimosine, which delays the cell cycle in G1. We find that in mimosine, centrosome duplication does occur, albeit slowly compared with cells that progress into S-phase; centrosome duplication in mimosine-treated cells also proceeds in the absence of a rise in Cdk2 kinase activity normally associated with the G1/S transition. CHO cells arrested with mimosine can also assemble more than four centrioles (termed "centrosome amplification"), but the extent of centrosome amplification during prolonged G1 is decreased compared to cells that enter S-phase and activate the Cdk2-cyclin complex. Together, our results suggest a model, which predicts that entry into S-phase and the rise in Cdk2 activity associated with this transition are not absolutely required to initiate centrosome duplication, but rather, serve to entrain the centrosome reproduction cycle with cell cycle progression.

Long-term outcomes of cardiac pacing in adults with congenital heart disease.

OBJECTIVES: The purpose of this retrospective study was to define long-term outcomes after pacemaker therapy in adults with congenital heart disease (CHD). BACKGROUND: Adults with CHD represent a unique and expanding population. Many will require pacemaker or implantable defibrillator therapy, with a lifelong need for re-intervention and follow-up. They pose technical and management challenges not encountered in other groups receiving pacing, and the complication and re-intervention rates specific to this population are not well-defined. METHODS: We reviewed outcomes of 168 adults with CHD, 89 females, mean age 40 years, in whom a pacemaker or anti-tachycardia device was implanted. RESULTS: Mean age at implant was 28 years with mean pacing duration 11 years at follow-up (range, 0.5 to 38.0). Seventy-two (42%) received initial dual-chamber devices and remained in this mode, while 23 (14%) went from ventricular to dual-chamber pacing in follow-up. Initial mode of pacing did not have a significant effect on subsequent atrial arrhythmia. Patients receiving an initial epicardial system were younger than those paced endocardially (17 +/- 12 years vs. 35 +/- 16 years, p < 0.001) and more likely to undergo re-intervention (p = 0.019). Difficulty with vascular access was encountered in 25 patients (15%), while 45 (27%) experienced lead-related complications. No significant predictors of lead complications were identified. CONCLUSIONS: Lead complications were not significantly different for epicardial versus endocardial, nor physiologic versus ventricular pacing, but a trend toward improved lead survival in patients receiving endocardial leads at first implant was observed. Adults with CHD remain at risk for atrial arrhythmias regardless of pacing mode.