Investigation of cell-accelerated corrosion (CAC) on the CoCrMo alloy with segregation banding: Hip implant applications.

Metal alloy microstructure plays a crucial role in corrosion associated with total hip replacement (THR). THR is a prominent strategy that uses metal implants such as cobalt-chromium-molybdenum (CoCrMo) alloys due to their advantageous biological and mechanical properties. Despite all benefits, these implants undergo corrosion and wear processes in-vivo in a synergistic manner called tribocorrosion. Also, the implant retrieval findings reported that fretting corrosion occurred in-vivo, evidenced by the damage patterns that appeared on the THR junction interfaces. There is no scientific data on the studies reporting the fretting corrosion patterns of CoCrMo microstructures in the presence of specific biological treatments to date. In the current study, Flat-on-flat fretting corrosion set-up was customized and used to study the tribocorrosion patterns of fretting corrosion to understand the role of alloy microstructure. Alloy microstructural differences were created with the implant stock metal's longitudinal and transverse cutting orientations. As a result, the transverse created the non-banded, homogenous microstructure, whereas the longitudinal cut resulted in the banded, non-homogenous microstructure on the surface of the alloy (in this manuscript, the terms homogenous and banded were used). The induced currents were monitored using a three-electrode system. Three different types of electrolytes were utilized to study the fretting corrosion patterns with both homogeneous and banded microstructures: 1. Control media 2. Spent media (the macrophage cell cultured media) 3. Challenged media (media collected after the macrophage was treated with CoCrMo particles). From the electrochemical results, in the potentiostat conditions, the banded group exhibited a higher induced current in both challenged and spent electrolyte environments than in control due to the synergistic activity of CoCrMo particles and macrophage demonstrating more corrosion loss. Additionally, both Bode and Nyquist plots reported a clear difference between the banded and homogeneous microstructure, especially with challenged electrolytes becoming more corrosion-resistant post-fretting than pre-fretting results. The banded microstructure showed a unique shape of the fretting loop, which may be due to tribochemical reactions. Therefore, from the electrochemical, mechanical, and surface analysis data results, the transverse/homogenous/non-banded alloy microstructure groups show a higher resistance to fretting-corrosion damage.

Proteomic-based electrochemical non-invasive biosensor for early breast cancer diagnosis.

Breast cancer is the second highest cause of cancer-related mortality in women worldwide and in the United States, accounting for around 571,000 deaths per year. Early detection of breast cancer increases treatment results and the possibility of a cure. While existing diagnostic modalities such as mammography, ultrasound, and biopsy exist, some are prohibitively expensive, uncomfortable, time-consuming, and have limited sensitivity, necessitating the development of a cost-effective, rapid, and highly sensitive approach such as an electrochemical biosensor. Our research focuses on detecting breast cancer patients using the ECM1 biomarker, which has higher expression in synthetic urine. Our study has two primary objectives: (i) Diverse ECM1 protein concentrations are measured using electrochemical impedance spectroscopy and ELISA. Establishing a standard curve for the electrochemical biosensor by calibrating ECM-1 protein levels using electrochemical impedance spectroscopy. (ii) Validation of the effectiveness of the electrochemical biosensor. This aim entails testing the unknown concentration of ECM1 in the synthetic urine to ensure the efficiency of the biosensor to detect the biomarker in the early stages. The results show that the synthetic urine solution's ECM-1 detection range ranges from 1 pg/ml to 500 ng/ml. This shows that by detecting changes in ECM-1 protein levels in patient urine, the electrochemical biosensor can consistently diagnose breast cancer in its early stages or during increasing recurrence. Our findings highlight the electrochemical biosensor's efficacy in detecting early-stage breast cancer biomarkers (ECM-1) in patient urine. Further studies will be conducted with patient samples and develop handheld hardware for patient usage.

Methotrexate and sulforaphane loaded PBA-G5-PAMAM dendrimers as a combination therapy for anti-inflammatory response in an intra-articular joint arthritic animal model.

L-sulforaphane (LSF), a natural product developed from cruciferous vegetables, have shown potent anti-inflammatory effect in cancer as well as arthritis. However, the stable delivery of LSF remains a major challenge. Methotrexate (MTX) is currently the first line treatment for managing RA and is most effective in patients when used in combination with other anti-inflammatory or anti-rheumatic drugs. Here we developed phenylboronic acid-PAMAM dendrimer (PBA-G5D) nanoparticles conjugated MTX (MTX-PBA-G5D), and L-sulforaphane (LSF/PBA-G5D) loaded dendrimers. The MTX and LSF drug loading and release kinetics was analyzed using HPLC. The lipopolysaccharide (LPS) stimulated macrophages were treated with the formulations to study the inflammatory response in vitro. For in vivo studies, arthritis was induced in five-week-old female Wistar rats, and the MTX- and LSF/PBA-G5-D were injected via intra-articular injection for treatment and the arthritis reduction was scored by weight, knee diameter, and serum cytokine level measurement. The average size of the drug-nanoparticle conjugates ranged from 135 to 250 nm, with mostly cationic surface charges. The encapsulation efficiency of the drugs to the modified dendrimer was more than 60% with a slow release of drugs from the nanoparticles within 24 h at pH 7.4. Drugs in the nanoparticle formulation were biocompatible, with promising anti-inflammatory effects in vitro against LPS-activated murine macrophages. Further in vivo studies on arthritis induced female Wistar rats, revealed significant anti-arthritic effects based on the arthritic scoring from the knee diameter reading, and anti-inflammatory effects based on the serum cytokine levels. This study provides a promising strategy for utilizing PAMAM dendrimers as a nanocarrier for LSF delivery for RA therapy.

Current advancements in bio-ink technology for cartilage and bone tissue engineering.

In tissue engineering, the fate of a particular organ/tissue regeneration and repair mainly depends on three pillars - 3D architecture, cells used, and stimulus provided. 3D cell supportive structure development is one of the crucial pillars necessary for defining organ/tissue geometry and shape. In recent years, the advancements in 3D bio-printing (additive manufacturing) made it possible to develop very precise 3D architectures with the help of industrial software like Computer-Aided Design (CAD). The main requirement for the 3D printing process is the bio-ink, which can act as a source for cell support, proliferation, drug (growth factors, stimulators) delivery, and organ/tissue shape. The selection of the bio-ink depends upon the type of 3D tissue of interest. Printing tissues like bone and cartilage is always challenging because it is difficult to find printable biomaterial that can act as bio-ink and mimic the strength of the natural bone and cartilage tissues. This review describes different biomaterials used to develop bio-inks with different processing variables and cell-seeding densities for bone and cartilage 3D printing applications. The review also discusses the advantages, limitations, and cell bio-ink compatibility in each biomaterial section. The emphasis is given to bio-inks reported for 3D printing cartilage and bone and their applications in orthopedics and orthodontists. The critical/important performance and the architectural morphology requirements of desired bone and cartilage bio-inks were compiled in summary.

Demonstration of the DNA Fiber Assay for Investigating DNA Damage and Repair Dynamics Induced by Nanoparticles.

Nanomaterial exposure can cause replication stress and genomic instability in cells. The degree of instability depends on the chemistry, size, and concentration of the nanomaterials, the time of exposure, and the exposed cell type. Several established methods have been used to elucidate how endogenous/exogenous agents impact global replication. However, replicon-level assays, such as the DNA fiber assay, are imperative to understand how these agents influence replication initiation, terminations, and replication fork progression. Knowing this allows one to understand better how nanomaterials increase the chances of mutation fixation and genomic instability. We used RAW 264.7 macrophages as model cells to study the replication dynamics under graphene oxide nanoparticle exposure. Here, we demonstrate the basic protocol for the DNA fiber assay, which includes pulse labeling with nucleotide analogs, cell lysis, spreading the pulse-labeled DNA fibers onto slides, fluorescent immunostaining of the nucleotide analogs within the DNA fibers, imaging of the replication intermediates within the DNA fibers using confocal microscopy, and replication intermediate analysis utilizing a computer-assisted scoring and analysis (CASA) software.

In vitro assessment of varying peptide surface density on the suppression of angiogenesis by micelles displaying αvß3 blocking peptides.

Ligand targeted therapy (LTT) is a precision medicine strategy that can selectively target diseased cells while minimizing off-target effects on healthy cells. Integrin-targeted LTT has been developed recently for angiogenesis-related diseases. However, the clinical success is based on the optimal design of the nanoparticles for inducing receptor clustering within the cell membrane. The current study focused on determining the surface density of Ser-Asp-Val containing anti-integrin heptapeptide on poly (ethylene glycol)-b-poly(propylene sulfide) micelles (MC) required for anti-angiogenic effects on HUVECs. Varying peptide density on PEG-b-PPS/Pep-PA MCs (Pep-PA-Peptide-palmitoleic acid) was used in comparison to a random peptide (SGV) and cRGD (cyclic-Arginine-Glycine-Aspartic acid) construct at 5%-density on MCs. Immunocytochemistry using CD51/CD31 antibody was performed to study the integrin blocking by MCs. In addition, the expression of VWF and PECAM-1, cell migration and tube formation was evaluated in the presence of PEG-b-PPS/Pep-PA MCs. The results show PEG-b-PPS/SDV-PA MCs with 5%-peptide density to achieve significantly higher αvß3 blocking compared to random peptide as well as cRGD. In addition, αvß3 blocking via MCs further reduced the expression of vWF and PECAM-1 angiogenesis protein expression in HUVECs. Although a significant level of integrin blocking was observed for 1%-peptide density on MCs, the cell migration and tube formation were not significantly affected. In conclusion, the results of this study demonstrate that the peptide surface density on PEG-b-PPS/Pep-PA MCs has a significant impact in integrin blocking as well as inhibiting angiogenesis during LTT. The outcomes of this study provides insight into the design of ligand targeted nanocarriers for various disease conditions.

Hip implant modular junction: The role of CoCrMo alloy microstructure on fretting-corrosion.

Cobalt-chromium-molybdenum (CoCrMo) alloy is one of the most used metals in total hip replacement (THR) due to the alloy's superior corrosion qualities and biocompatibility. Over time these prostheses may undergo wear and corrosion processes in a synergistic process known as tribocorrosion. Implant retrieval studies have shown that damage patterns on THR modular junction surfaces indicating specifically in vivo fretting-corrosion to take place. To date, there have been no studies on the fretting-corrosion behaviors of CoCrMo alloy under the consideration of specific microstructural features. A custom-built flat-on-flat fretting-corrosion setup was utilized to test the synergistic tribocorrosion behavior of fretting-corrosion. The difference in microstructure was generated through the cutting orientations of the transverse and the longitudinal direction of the bar stock material, where the longitudinal cut exhibits a characteristic banded microstructure (banded group) and the transverse cut a homogenous microstructure (unbanded group). A three-electrode system was employed to monitor the induced currents. Two different types of electrolytes were used in the current study: 1. Bovine calf serum (BCS-30 g/L protein) (normal conditions) 2. BCS with Lipopolysaccharide (LPS, 0.15 µg/ml) (simulated infectious conditions). In the free potential mode, banded samples showed an increased potential compared to the unbanded samples. In potentiostatic conditions, the banded group also exhibited a higher induced current in both electrolyte environments, indicating more corrosion loss. Both Nyquist and Bode plots showed both orientations of metal becoming more corrosion resistant post-fretting when compared to pre-fretting data. The longitudinal group at OCP demonstrated a unique shape of the fretting-loop, which might be related to tribochemical reactions. Based on the mechanical, electrochemical, and surface characterization data, the transverse group (unbanded) microstructures demonstrates a higher resistance to fretting-corrosion damage.

Amniotic growth factors enhanced human pre-adipocyte cell viability and differentiation under hypoxia.

One of the major drawbacks associated with autologous fat grafting is unpredictable graft retention. Various efforts to improve the survivability of these cells have been explored, but these methods are time-consuming, complex, and demand significant technical skill. In our study, we examine the use of cryopreserved amniotic membrane as a source of exogenous growth factors to improve adipocyte survivability under normal and hypoxic conditions. Human primary preadipocytes were cultured in a gelatin-ferulic acid (Gtn-FA) hydrogel with variable oxygen concentration and treated with amniotic membrane-derived condition medium (CM) for 7 days. This hydrogel provides a hypoxic environment and also creates a 3D cell culture to better mimic recipient site conditions. The O2 concentration in the hydrogel was measured by electron paramagnetic resonance oxygen imaging (EPROI). The conjugation of FA was confirmed by FTIR and NMR spectroscopy. The cell viability and adipocyte differentiation were analyzed by alamarBlue™ assay, Oil Red O staining, and RT-qPCR. The expression of genes: Pref-1, C/EBP ß, C/EBP α, PPAR-Æ´, SLC2A4, and VEGF-A were quantified. The cell viability results show that the 50% CM showed significantly higher cell pre-adipocyte cell viability. In addition, compared to normal conditions, hypoxia/CM provided higher PPAR-Æ´ (p < .05), SLC2A4, and VEGF-A (p < .05) (early and terminal differentiating markers) mRNA expression. This finding demonstrates the efficacy of amniotic CM supplementation as a novel way to promote adipocyte survival and retention via the expression of key gene markers for differentiation and angiogenesis.

Total hip replacement monitoring: numerical models for the acoustic emission technique.

Any mechanical instability associated with total hip replacement (THR) excites elastic waves with different frequencies and propagates through the surrounding biological layers. Using the acoustic emission (AE) technique as a THR monitoring tool provides valuable information on structural degradations associated with these implants. However, several factors can compromise the reliability of the signals detected by AE sensors, such as attenuation of the detected signal due to the presence of biological layers in the human body between prosthesis (THR) and AE sensor. The main objective of this study is to develop a numerical model of THR that evaluates the impact of biological layer thicknesses on AE signal propagation. Adipose tissue thickness, which varies the most between patients, was modeled at two different thicknesses 40 mm and 70 mm, while the muscle and skin thicknesses were kept to a constant value. The proposed models were tested at different micromotions of 2 µm, 15-20 µm at modular junctions, and different frequencies of 10-60 kHz. Attenuation of signal is observed to be more with an increase in the selected boundary conditions along with an increase in distance the signals propagate through. Thereby, the numerical observations drawn on each interface helped to simulate the effect of tissue thicknesses and their impact on the attenuation of elastic wave propagation to the AE receiver sensor.

Corrosion Behavior of Selective Laser Melting (SLM) Manufactured Ti6Al4V Alloy in Saline and BCS Solution.

The frequency of surgeries involving the use of metal implants in orthopedic medicine to replace degenerative or fractured joints is increasing, and it is therefore important to optimize the lifespan and quality of these implants. Advances in additive manufacturing (AM), or 3D printing, are creating new opportunities to personalize implants in ways that reduce mechanical stress at the joint implant interface and improve bone ingrowth and implant stability; however, it is not well understood if and to what degree the AM process alters the corrosion behavior of the materials it produces. In this study, six Ti6Al4V prints manufactured via a selective laser melting (SLM) method were examined regarding their corrosion behavior in both saline and bovine calf serum (BCS) solutions. Ecorr and Icorr values were comparable between the CM-Ti6Al4V control and SLM-EDM surfaces; however, SLM surfaces were found to have more narrow passivation behavior evidenced by significant decreases in Epass values relative to CM-Ti6Al4V. We believe this is a consequence of microstructural differences between CM-Ti6Al4V and SLM-Ti6Al4V. Specifically, the SLM-Ti6Al4V demonstrated a dominant α' martensitic microstructure and decreased vanadium-rich ß-phase. BCS solution had a detrimental effect on potential parameters, Ecorr and OCP, decreasing these values relative to their saline counterparts. Increased surface roughness of the SLM-printed surface seemed to amplify the effects of the BCS solution. Furthermore, modest decreases in Epass and Ipass were observed in BCS solution, suggesting that the presence of protein may also interfere with passivation behavior. These findings have implications for how SLM-Ti6Al4V implants will perform in vivo and could possibly influence implant longevity and performance.

Dynamic microfluidic bioreactor-Hip simulator (DMBH) system for implant toxicity monitoring.

The generation of degradation products (DPs) like ions and organo-metallic particles from corroding metallic implants is an important healthcare concern. These DPs generate local and systemic toxicity. The impact on local toxicity is well documented, however, little is known about systemic toxicity. This is mainly due to the limited scope of the current microtiter plate-based (static) toxicity assay techniques. These methods do not mimic the systemic (dynamic) conditions. In this study, it is hypothesized that DPs incubated with cells in static conditions might provide improper systemic toxicity results, as there is no movement mimicking the blood circulation around cells. This study reports the development of a three-chambered prototype microfluidic system connected to the operational hip implant simulator to test the cellular response induced by the DPs. This setup is called a dynamic microfluidic bioreactor-hip simulator system. We hypothesize that a dynamic microfluidic system will provide a realistic toxicology response induced by DPs than a static cell culture plate. To prove the hypothesis, Neuro2a (N2a) cells were used as representative cells to study systemic neurotoxicity by the implant DPs. The microfluidic bioreactor system was validated by comparing the cell toxicity against the traditional static system and using COMSOL modeling for media flow with DPs. The hip implant simulator used in this study was a state-of-the-art sliding hip simulator developed in our lab. The results suggested that static toxicity was significantly more compared to dynamic microfluidic-based toxicity. The newly developed DMBH system tested for in situ systemic toxicity on N2a cells and demonstrated very minimum toxicity level (5.23%) compared to static systems (31.16%). Thus, the new DMBH system is an efficient tool for in situ implant metal systemic toxicity testing.

The role of Vitamin E in hip implant-related corrosion and toxicity: Initial outcome.

In orthopedic healthcare, Total Hip Replacement (THR) is a common and effective solution to hip-related bone and joint diseases/fracture; however, corrosion of the hip implant and the release of degradation metal ions/particles can lead to early implant failure and pose potential toxicity risk for the surrounding tissues. The main objective of this work was to investigate the potential role of Vitamin E to minimize corrosion-related concerns from CoCrMo hip implants. The study focused on two questions (i) Can Vitamin E inhibit CoCrMo corrosion? and (ii) Does Vitamin E moderate the toxicity associated with the CoCrMo implant particles? In the study (i) the electrochemical experiments (ASTM G61) with different concentrations of Vitamin E (1, 2, 3 mg/ml against the control) were performed using normal saline and simulated synovial fluid (Bovine calf serum-BCS, 30 g/L protein, pH 7.4) as electrolytes. The polished CoCrMo disc (Ra 50 nm) was the working electrode. The findings suggested that both Vitamin E-Saline (45 ± 0.9%) and Vitamin E-BCS (91 ± 3%) solutions protected against implant corrosion at a Vitamin E concentration of 3 mg/ml, but Vitamin E-BCS showed protection at all Vitamin E (1-3 mg/ml) concentration levels. These results suggested that the Vitamin E and the protein present in the BCS imparted additive effects towards the electrochemical inhibition. In the study (ii) the role of Vitamin E in cytotoxicity inhibition was studied using a mouse neuroblastoma cell line (N2a) for CoCrMo particles and Cr ions separately. The CoCrMo particles were generated from a custom-built hip simulator. The alamarBlue assay results suggested that Vitamin E provides significant protection (85% and 75% proliferation) to N2a cells against CoCrMo particles and Cr ions, respectively at 1 µg/ml concentration, as compared to the control group. However, the results obtained from ROS expression and DNA fiber staining suggest that Vitamin E is only effective against CoCrMo degradation particles and not against Cr ions. In summary, the findings show that Vitamin E can minimize the corrosion processes and play a role in minimizing the potential toxicity associated with implants.

Systemic toxicity eliciting metal ion levels from metallic implants and orthopedic devices - A mini review.

The metal/metal alloy-based implants and prostheses are in use for over a century, and the rejections, revisions, and metal particle-based toxicities were reported concurrently. Complications developed due to metal ions, metal debris, and organo-metallic particles in orthopedic patients have been a growing concern in recent years. It was reported that local and systemic toxicity caused by such released products from the implants is one of the major reasons for implant rejection and revision. Even though the description of environmental metal toxicants and safety limits for their exposure to humans were well established in the literature, an effort was not adequately performed in the case of implant-based metal toxicology. Since the metal ion concentration in serum acts as a possible indicator of the systemic toxicity, this review summarizes the reported human serum safe limits, toxic limits, and concentration range (µg/L, ppb, etc.) for mild to severe symptoms of six (cardiac, hepatic, neuro, nephron, dermal and endocrine) systemic toxicities for twelve most commonly used metallic implants. It also covers the widely used metal ion quantification techniques and systemic toxicity treatments reported.

The role of fretting-frequency on the damage modes of THR modular junction: In-vitro study.

According to the National Center for Health Statistics, currently, more than 250,000 total hip replacements annually in the US alone, with an estimated increase to 500,000 by the year 2030. The usage of tapered junctions between the femoral neck and head gives the surgeon flexibility in implant assembly. However, these modular junctions are subjected to micro-motion that may cause chemical and fretting-corrosion at the modular junction. Therefore, it is imperative to study these forces to mitigate their effects. The current study aims to understand the effects of fretting-corrosion as a function of fretting frequencies caused by common physical activities in an in-vitro model of hip modular junctions. The fretting system has a tribological contact condition of flat-on-flat, mounted to a load frame. CoCrMo pins were polished and immersed in a synovial fluid-like electrolyte solution (Bovine calf serum 30 g/l). Electrochemical measurements were made using a potentiostat. Samples then undergo 3600 cycles at 50 µm (to simulate gross slips), with a horizontal load at 200 N, and a frequency of 0.5 Hz, 0.7 Hz, 1 Hz, and 1.5 Hz to simulate Sit Down-Stand Up, Stair Climb, Walking, and Jogging, respectively. Worn surfaces were then examined under optical and scanning electron microscopy. The evolution of free potential as a function of time for tested frequencies shows the initial potential drop and stabilized trend in the potential evolution. The sample group at a higher frequency displays a higher tendency of corrosion than a lower frequency; however, the dissipation energy decreases as a function of fretting frequency. Both electrochemical and mechanical responses correlate to the variation in the fretting frequencies. Organometallic complexes were found on the surfaces of the samples that were subjected to a slower frequency of fretting, whereas mechanical grooving was noticed on samples with a faster frequency. Hence, these preliminary studies suggest that implant failure rates may be altered based on fretting-frequencies induced by physical activity. Further studies will be required to verify the findings and explore the potential role of fretting frequency in the damage modes of the modular junction.

Non-invasive early detection of failure modes in total hip replacements (THR) via acoustic emission (AE).

Total hip replacements (THR) are becoming an common orthopedic surgucal procedure in the United States (332 K/year in 2017) to relieve pain and improve the mobility of those that are affected by osteoarthritis, ankylosing spondylitis, or injury. However, complications like tribocorrosion, or material degradation due to friction and corrosion, may result in THR failure. Unfortunately, few strategies to non-invasively diagnose early-stage complications are reported in literature, leading to implant complications being detected after irreversible damage. Therefore, the main objective of this study proposes the utilization of acoustic emission (AE) to continuously monitor implant materials, CoCrMo and Ti6Al4V, and identify degradations formed during cycles of sleeping, standing, and walking by correlating them to potential and friction coefficient behavior. AE activity detected from the study correlates with the friction coefficient and open-circuit potential observed during recreated in-vitro standing, walking, and sleeping cycles. It was found that the absolute energy level obtained from AE increased as the friction coefficient increased, potential decreased, and wear volume loss increased. Through the results, higher friction coefficient and AE activity were observed in Ti6Al4V alloys while there was also a significant drop in potential, indicating increased tribocorrosion activity. Therefore, AE can be utilized to predict material degradations as a non-invasive method based on the severity of abnormality of the absolute energy and hits emitted. The correlation between potential, friction coefficient, and AE activity was further confirmed through profilometry which showed more material degradation in Ti6Al4V than CoCrMo. Through these evaluations, it was demonstrated that AE could be utilized to identify the deformations and failure modes of implant materials caused by tribocorrosion.

Implant-derived CoCrMo alloy nanoparticle disrupts DNA replication dynamics in neuronal cells.

The complexity of cobalt-chromium-molybdenum (CoCrMo) nanoparticles generated from the hip modular taper interfaces resulted in inconclusive outcomes on the level of toxicity in orthopedic patients. We used a hip simulator to generate physiologically relevant CoCrMo degradation products (DPs) to demonstrate the variation in the level of toxicity in neurons in comparison to processed degradation products (PDPs). The study outcomes indicate that DP induces a higher level of DNA damage in the form of double- and single-stranded DNA breaks and alkaline labile DNA adducts versus PDPs. The scientific advancements of this study are the following: (i) how DPs mimic more closely to the implant debris from hip implants in terms of bioactivity, (ii) how hip implant debris causes local and systemic issues, and (iii) methods to augment the biologic impact of implant debris. We discovered that DP is bioactive compared with PDP, and this should be considered in the toxicity evaluation related to implants. • The physicochemical characteristics of the CoCrMo is a major factor to consider for implant-related cytotoxicity or genotoxicity experimental design. • Elevated levels of intracellular ROS induced by the physiologically relevant wear particle are detrimental to the neuronal cells. • The DP can induce variation in DNA replication dynamics compared to PDP.

High Density Display of an Anti-Angiogenic Peptide on Micelle Surfaces Enhances Their Inhibition of αvß3 Integrin-Mediated Neovascularization In Vitro.

Diabetic retinopathy (DR), Retinopathy of Pre-maturity (ROP), and Age-related Macular Degeneration (AMD) are multifactorial manifestations associated with abnormal growth of blood vessels in the retina. These three diseases account for 5% of the total blindness and vision impairment in the US alone. The current treatment options involve heavily invasive techniques such as frequent intravitreal administration of anti-VEGF (vascular endothelial growth factor) antibodies, which pose serious risks of endophthalmitis, retinal detachment and a multitude of adverse effects stemming from the diverse physiological processes that involve VEGF. To overcome these limitations, this current study utilizes a micellar delivery vehicle (MC) decorated with an anti-angiogenic peptide (aANGP) that inhibits αvß3 mediated neovascularization using primary endothelial cells (HUVEC). Stable incorporation of the peptide into the micelles (aANGP-MCs) for high valency surface display was achieved with a lipidated peptide construct. After 24 h of treatment, aANGP-MCs showed significantly higher inhibition of proliferation and migration compared to free from aANGP peptide. A tube formation assay clearly demonstrated a dose-dependent angiogenic inhibitory effect of aANGP-MCs with a maximum inhibition at 4 µg/mL, a 1000-fold lower concentration than that required for free from aANGP to display a biological effect. These results demonstrate valency-dependent enhancement in the therapeutic efficacy of a bioactive peptide following conjugation to nanoparticle surfaces and present a possible treatment alternative to anti-VEGF antibody therapy with decreased side effects and more versatile options for controlled delivery.

In Vitro Evidence for Cell-Accelerated Corrosion Within Modular Junctions of Total Hip Replacements.

Corrosion at modular junctions of total hip replacement (THR) remains a major concern today. Multiple types of damage modes have been identified at modular junctions, correlated with different corrosion characteristics that may eventually lead to implant failure. Recently, within the head-taper region of the CoCrMo retrieval implants, cell-like features and trails of etching patterns were observed that could potentially be linked to the involvement of cells of the periprosthetic region. However, there is no experimental evidence to corroborate this phenomenon. Therefore, we aimed to study the potential role of periprosthetic cell types on corrosion of CoCrMo alloy under different culture conditions, including the presence of CoCrMo wear debris. Cells were incubated with and without CoCrMo wear debris (obtained from a hip simulator) with an average particle size of 119 ± 138 nm. Electrochemical impedance spectroscopy (EIS) was used to evaluate the corrosion tendency, corrosion rate, and corrosion kinetics using the media after 24 h of cell culture as the electrolyte. Results of the study showed that there was lower corrosion resistance (p < 0.02) and higher capacitance (p < 0.05) within cell media from macrophages challenged with particles when compared with the other media conditions studied. The potentiodynamic results were also in agreement with the EIS values, showing significantly higher corrosion tendency (low Ecorr ) (p < 0.0001) and high Icorr (p < 0.05) in media from challenged macrophages compared with media with H2 O2 solution. Overall, the study provides in vitro experimental evidence for the possible role of macrophages in altering the chemical environment within the crevice and thereby accelerating corrosion of CoCrMo alloy. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:393-404, 2020.

Wear particles induce a new macrophage phenotype with the potential to accelerate material corrosion within total hip replacement interfaces.

Evidence that macrophages can play a role in accelerating corrosion in CoCrMo alloy in total hip replacement (THR) interfaces leads to questions regarding the underlying cellular mechanisms and immunological responses. Hence, we evaluated the role of macrophages in corrosion processes using the cell culture supernatant from different conditions and the effect of wear particles on macrophage dynamics. Monocytes were exposed to CoCrMo wear particles and their effect on macrophage differentiation was investigated by comparisons with M1 and M2 macrophage differentiation. Corrosion associated macrophages (MCA macrophages) exhibited upregulation of TNF-α, iNOS, STAT-6, and PPARG and down-regulation of CD86 and ARG, when compared to M1 and M2 macrophages. MCA cells also secreted higher levels of IL-8, IL-1ß, IL-6, IL-10, TNF-α, and IL-12p70 than M1 macrophages and/or M2 macrophages. Our findings revealed variation in macrophage phenotype (MCA) induced by CoCrMo wear particles in generating a chemical environment that induces cell-accelerated corrosion of CoCrMo alloy at THR modular interfaces. STATEMENT OF SIGNIFICANCE: Fretting wear and corrosion within the implant's modular taper junction are prominent causes of implant failure, as they promote the release of corrosion products and subsequent development of adverse local tissue reactions. Being a multifactorial process, several in vitro models have been developed to recreate the in vivo corrosion process, often summarized as mechanically-assisted crevice corrosion. Considering the excellent corrosion properties of CoCrMo alloy, the severity of chemically-generated damage observed at the modular interface has been surprising and poorly understood. The aim of the current study is to provide a better understanding of macrophages and their plasticity at the THR taper interface when they encounter wear debris from CoCrMo alloy. This is a preliminary study along the path towards determining the mechanism(s) of CAC.