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.

Regenerative Medicine Strategies in Biomedical Implants.

PURPOSE OF REVIEW: Recently, significant progress has been made in the research related to regenerative medicine. At the same time, biomedical implants in orthopedics and dentistry are facing many challenges and posing clinical concerns. The purpose of this chapter is to provide an overview of the clinical applications of current regenerative strategies to the fields of dentistry and orthopedic surgery. The main research question in this review is: What are the major advancement strategies in regenerative medicine that can be used for implant research? RECENT FINDINGS: The implant surfaces can be modified through patient-specific stem cells and plasma coatings, which may provide methods to improve osseointegration and sustainability of the implant. Overall understanding from the review suggesting that the outcome from the studies could lead to identify optimum solutions for many concerns in biomedical implants and even in drug developments as a long-term solution to orthopedic and dental patients.

Serum Metal Levels in Maxillofacial Reconstructive Surgery Patients: A Pilot Study.

PURPOSE: The aim of this pilot study was to assay metal concentrations in the serum of patients who had undergone dental implant placement, orthognathic surgery using rigid metal fixation plates and screws, and total temporomandibular joint replacement (TMJ TJR). MATERIALS AND METHODS: Thirty patients were identified and included in this pilot study. Sixteen patients (9 men and 8 women), with an average age of 44 years (range, 19 to 79 yr), provided informed consent to participate and were divided into 3 study groups with 4 patients in each (group 1, orthognathic surgery; group 2, TMJ TJR; and group 3, dental implant placement). A control group consisted of volunteers without any implanted metallic devices. Blood samples for serum metal analysis were obtained and analyzed in accordance with the standardized collection and testing protocols used at the Trace Metal Analysis Laboratory of the Department of Orthopedic Surgery at the Rush University Medical Center (Chicago, IL). RESULTS: All control participants had levels below the normal reference range for all serum markers assessed. In the orthognathic group, 1 patient had an increased serum cobalt level. In the TMJ TJR group, 1 patient had an increased serum cobalt level and another patient had an increased serum chromium level. In the dental implant group, 1 patient had an increased serum titanium level and another had increased serum levels of titanium and chromium. CONCLUSIONS: This is the first study to report on the release of metal into the bloodstream in patients with different maxillofacial implanted metallic objects. The results raise questions regarding the types and magnitude of metal released from maxillofacial reconstruction devices and their potential long-term local and systemic effects. Future large-scale prospective studies involving serial measurements in homogeneous groups of patients could further elucidate the impact of these findings.

Synthesis and Evaluation of a Sodium Alginate-4-Aminosalicylic Acid Based Microporous Hydrogel for Potential Viscosupplementation for Joint Injuries and Arthritis-Induced Conditions.

A microporous hydrogel was developed using sodium alginate (alg) and 4-aminosalicylic acid (4-ASA). The synthesized hydrogel was characterized using various analytical techniques such as Fourier transform infrared spectroscopy (FTIR), Carbon-13 nuclear magnetic resonance (13C-NMR), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and differential scanning calorimetry (DSC). Additonal carboxyl and hydroxyl functional groups of 4-ASA provided significant lubrication and stress-triggered sol-gel transition to the conjugated hydrogel. In addition, cytotoxicity analysis was undertaken on the conjugated hydrogel using human dermal fibroblast-adult (HDFa) cells, displaying non-toxic characteristics. Drug release profiles displaying 49.6% in the first 8 h and 97.5% within 72 h, similar to the native polymer (42.8% in first 8 h and 90.1% within 72 h). Under applied external stimuli, the modified hydrogel displayed significant gelling properties and structure deformation/recovery behaviour, confirmed using rheological evaluation (viscosity and thixotropic area of 8095.3 mPas and 26.23%, respectively). The modified hydrogel, thus, offers great possibility for designing smart synovial fluids as a biomimetic aqueous lubricant for joint-related injuries and arthritis-induced conditions. In addtion, the combination of thixotropy, non-toxicity, and drug release capabilities enables potential viscosupplementation for clinical application.

Implantable and transdermal polymeric drug delivery technologies for the treatment of central nervous system disorders.

The complexity of the brain and the membranous blood-brain barrier (BBB) has proved to be a significant limitation to the systemic delivery of pharmaceuticals to the brain rendering them sub-therapeutic and ineffective in the treatment of neurological diseases. Apart from this, lack of innovation in product development to counteract the problem is also a major contributing factor to a poor therapeutic outcome. Various innovative strategies show potential in treating some of the neurological disorders; however, drug delivery remains the most popular. To attain therapeutic drug levels in the central nervous system, large, intolerable systemic doses are generally administered. The major factors responsible for the success maintenance therapy of neurological diseases included controlled and sustained release of neurotherapeutics, reduced frequency of administration, higher bioavailability, and patient compliances. Conventional oral or injectable formulations cannot satisfy all the requirements in many circumstances. This article reviews the therapeutic implantable polymeric and transdermal devices employed in an attempt to effectively achieve therapeutic quantities of drug across the BBB over a prolonged period, to improve patient disease prognosis.

An electro-conductive fluid as a responsive implant for the controlled stimuli-release of diclofenac sodium.

The purpose of this study was to develop an electro-responsive co-polymeric (ERP) implantable gel from polyethylene glycol (PEG), sodium polystyrene sulphonate (NaPss), polyvinyl alcohol (PVA), and diethyl acetomidomalonate (DAA) for electro-liberation of the model drug diclofenac sodium. Various physicochemical and physicomechanical characterization tests were undertaken on the synthesized drug-free gel (ERP G1) and drug-loaded gel (ERP G2). The ability of the gel to release diclofenac sodium following electrical stimulation was evaluated using a galvanostat while Molecular Mechanics (MM) simulations were performed to elucidate the experimental mechanisms. A stable electro-active gel exhibiting superior cycling stability was produced with desirable rheological properties, rigidity (BHN = 35.4 N ± 0.33 N/mm2; resilience = 10.91 ± 0.11%), thermal properties (Tg ≈ 70 °C; Tc ≈ 200 °C) and homogeneous morphology. "ON-OFF" pursatile gradual drug release (37-94% from t30 min-t180 min) kinetics was observed upon applying electric stimulation intermittently, indicating that drug release from the gel was electrically controlled. Overall, the galvanometric and MM evaluation ascertained the suitability of the PEG/NaPss/PVA ERP-Gel for application as a subcutaneously injectable drug delivery implant.

Design of an Inflammation-Sensitive Polyelectrolyte-Based Topical Drug Delivery System for Arthritis.

The most successful treatment strategy for arthritis is intra-articular injections that are costly and have reduced patient compliance. The purpose of the current study was to develop an inflammation-sensitive system for topical drug administration. Multi-macromolecular alginate-hyaluronic acid-chitosan (A-H-C) polyelectrolyte complex nanoparticles, loaded with indomethacin were developed employing pre-gel and post-gel techniques in the presence of dodecyl-L-pyroglutamate (DLP). In addition to in vitro studies, in silico simulations were performed to affirm and associate the molecular interactions inherent to the formulation of core all-natural multi-component biopolymeric architectures composed of an anionic (alginate), a cationic (chitosan), and an amphi-ionic polyelectrolytic (hyaluronic acid) macromolecule. The results demonstrated that DLP significantly influenced the size of the synthesized nanoparticles. Drug-content analysis revealed higher encapsulation efficiency (77.3%) in the presence of DLP, irrespective of the techniques used. Moreover, in vitro drug release studies showed that indomethacin release from the nanosystem was significantly improved (98%) in Fenton's reagent. Drug permeation across a cellulose membrane using a Franz diffusion cell system showed an initial surge flux (0.125 mg/cm(-2)/h), followed by sustained release of indomethacin for the post-gel nanoparticles revealing its effective skin permeation efficiency. In conclusion, the study presents novel nanoparticles which could effectively encapsulate and deliver hydrophobic drugs to the target site, particularly for arthritis.

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.

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.

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.

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.

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.

Physicochemical and in-vitro biological analysis of bio-functionalized titanium samples in a protein-rich medium.

The long-term survivability of the implants is strongly influenced by the osseointegration aspects of the metal-bone interface. In this study, biological materials such as fibrinogen and fibrin are used to functionalize titanium surfaces to enhance the ability of implants to interact with human tissues for accelerated osseointegration. The biofunctionalized samples that were assessed by White Light Microscope, Scanning Electron Microscope and Water Contact Angle for surface properties proved samples etched with HF/HNO3 to be better than HCl/H2SO4 in terms of having optimum roughness and hydrophilicity for our further experiments. To further investigate the in vitro osseointegration of the biofunctionalized samples, Osteoblasts were cultured on the surfaces to assess cell proliferation, adhesion, gene expression as well as the mineralization process. Further bacterial adhesion (Enterococcus faecalis) and electrochemical evaluation of surface coating stability were carried out. Results of the study show that the biofunctionalized surfaces provided high cell proliferation, adherence, gene expression, and mineralization compared to other control surfaces hence proving them to have efficient and enhanced osseointegration. Also, bacterial adhesion studies show that there is no augmented growth of bacteria on the biofunctionalized samples in comparison to control surfaces. Electrochemical studies proved the existence of a stable protein layer on the bio functionalized surfaces. Such a method can reduce the time for osseointegration that can decrease risks in early failures of implants due to its enhanced hydrophilicity and cytocompatibility.

Differential toxicity of processed and non-processed states of CoCrMo degradation products generated from a hip simulator on neural cells.

Physico-chemical characteristics of the CoCrMo degradation products have played an important role in cytotoxicity and clinical complications on the orthopedic patients who have metal implants. Previous studies have limited reflection on the physicochemical characteristics of the degradation products generated in vivo, which are very different from individual metal particles and/or ions obtained from different commercial sources. In this study, we aimed to understand the differences in toxicity induced by the degradation products in as-synthesized form as well as those obtained after post-processing. The degradation products were generated using a hip-simulator by maintaining physiological conditions closer to in vivo and separated into two batches, one with processing by washing and drying called processed degradation products (PDP) and another batch as 'as-synthesized' degradation product (DP). We studied the dose-dependent toxicity response by neural cells derived from induced pluripotent stem cells. The results of the study show that as-synthesized DPs are more toxic to neural cells even at lower concentrations studied with evident low TC50 (1-5 µg/ml) concentrations compared to PDP (25 µg/ml). Flow cytometric analysis showed a significant (p<.01) increase in uptake of the particles after 24 h and corresponding ROS production in DP-treated cells. RT-PCR analysis of oxidative specific gene expression showed, elevated mRNA levels of NADPH oxidase-1, nuclear transcription factor, superoxide dismutase-2 and glutaredoxin-2 in DP-treated cells after 6 h. The results of the study provided a clear evidence of the differential response of neural cells on the degradation products as a function of concentrations and their chemical nature.

Total Eradication of Bacterial Infection in Root Canal Treatment: An Electrochemical Approach.

According to the American Association of Endodontists, currently 22.3 million endodontic procedures are being performed annually with the success rate of 70-95% and the average survival rate of the root canal procedure is approximately 67% after 5 years and 56% after 8 years. One of the major reason for the failure is relapse of infection. Hence, it is imperative to develop an assistive or alternative method to eradicate the bacterial infection effectively without affecting patient compliance. The application of electrochemistry has been used previously to disinfect catheters and implant disinfection. Hence, the aim of this study is to utilize the principles of electrochemistry to develop a microelectronic device to eradicate bacterial infection for root canal treatment. The electrochemical protocol includes open circuit potential (60 s) and potentiostatic scan at varying voltage (-9 to +2 V) at a different time duration (1-5 min). Enterococcus faecalis in the form of planktonic and biofilm was used in this study. After electrochemical treatment, the bacterial viability was evaluated using alamarBlue assay, colony forming units, confocal microscopy, and scanning electron microscopy. Cytotoxicity evoked by electrochemical voltage in comparison to NaOCl solution was performed using osteoblasts in 2D and 3D cell culture systems. The results of the study show that the application of -2 to +2 V at 1-5 min did not show any significant reduction in bacterial growth. However, the cathodic voltage of -9 V for 5 min showed a significant reduction (p < 0.001) in the bacterial count (80-95%). Similar results were obtained from biofilm study, which is more realistic to the in vivo condition. In contrast, the method did not induce cytotoxicity to the cells in 3D culture system (65% viability) in comparison to the highly toxic nature (0% viability) of NaOCl, indicating better patient compliance. Hence, the study provides supporting evidence to develop an electrochemically driven microelectronic device that can be a potential assistive dental instrument for endodontic procedures.

A composite chitosan-gelatin bi-layered, biomimetic macroporous scaffold for blood vessel tissue engineering.

A composite chitosan-gelatin macroporous hydrogel-based scaffold with bi-layered tubular architecture was engineered by solvent casting-co-particulate leaching. The scaffold constituted an inner macroporous layer concealed by a non-porous outer layer mimicking the 3D matrix of blood vessels with cellular adhesion and proliferation. The scaffold was evaluated for its morphological, physicochemical, physicomechanical and biodurability properties employing SEM, FTIR, DSC, XRD, porositometry, rheology and texture analysis. The fluid uptake and biodegradation in the presence of lysozymes was also investigated. Cellular attachment and proliferation was analysed using human dermal fibroblasts (HDF-a) seeded onto the scaffold and evaluated by MTT assay, SEM, and confocal microscopy. Results demonstrated that the scaffold had a desirable tensile strength=95.81±11kPa, elongation at break 112.5±13%, porosity 82% and pores between 100 and 230µm, 50% in vitro biodegradation at day 16 and proliferated fibroblasts over 20 days. These results demonstrate that scaffold may be an excellent tubular archetype for blood vessel tissue engineering.

A review of bioactive release from nerve conduits as a neurotherapeutic strategy for neuronal growth in peripheral nerve injury.

Peripheral nerve regeneration strategies employ the use of polymeric engineered nerve conduits encompassed with components of a delivery system. This allows for the controlled and sustained release of neurotrophic growth factors for the enhancement of the innate regenerative capacity of the injured nerves. This review article focuses on the delivery of neurotrophic factors (NTFs) and the importance of the parameters that control release kinetics in the delivery of optimal quantities of NTFs for improved therapeutic effect and prevention of dose dumping. Studies utilizing various controlled-release strategies, in attempt to obtain ideal release kinetics, have been reviewed in this paper. Release strategies discussed include affinity-based models, crosslinking techniques, and layer-by-layer technologies. Currently available synthetic hollow nerve conduits, an alternative to the nerve autografts, have proven to be successful in the bridging and regeneration of primarily the short transected nerve gaps in several patient cases. However, current research emphasizes on the development of more advanced nerve conduits able to simulate the effectiveness of the autograft which includes, in particular, the ability to deliver growth factors.