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1. NanoSUS (Ultra Fine Grained Stainless-Steel) for Percutaneous Spinal Injection

   Bahram Saleh, Caterina Bartomeu ( December 2019 , Tissue Engineering and Regenerative Medicine International Society) Start-up Competition, 2019)

   Vertebral augmentation, which includes vertebroplasty and kyphoplasty, is a minimally invasive procedure to relieve the back pain caused by vertebral compression fracture (VCF) and osteoporosis-related fractures. Vertebroplasty involves the introduction of a percutaneous needle into the vertebral body followed by an image-guided injection of cement directly into the vertebra. The market size for Vertebroplasty is expected to increase from $910 million in 2019, to $1500 million in 2024 with a CAGR of 8.4%, due to the growing demand by increasing geriatric population, rising incidence of spine illness and technological advancements in this field [1]. NanoSUS Bio-Tech (USA) was found based on a collaboration between Komatsuseiki Kosakusho Co., Ltd (Japan) and Northeastern University (NEU) in 2019 [2]. This company manufactures high strength ultra-fine-grained stainless steel with antimicrobial surface. However, nanoSUS has been developing ultra-fine-grained stainless steel since 2002 [3, 4]. Using this technology, we are able to control grain size without any change in chemical composition of commercially available FDA approved stainless steel. We are able to form this stainless steel into a vertebroplasty needle using our precision machining techniques at Komatsuseiki Kosakusho Co., Ltd. We believe this product will reduce the healthcare expenses for patients suffering from VCF due tomore »its cost-effective manufacturing process and antimicrobial surface, which reduces iatrogenic infections and shortens hospitalization time. We are going to submit a funding proposal to Center for Disruptive Musculoskeletal Innovation (CDMI) that will aid to expand the collaboration between Komatsuseiki Kosakusho Co., Ltd and NEU. The company will be established by financing from Komatsuseiki Kosakusho Co., Ltd (Japan), in Boston, MA. It will manage the collaborations between academia, marketing consultants and lawyers in US, and directors and researchers in Japan.« less
2. Developing Bioactive-Nanograined Metals With Bacterial Biofilm Inhibition Properties For Orthopedic Devices

   Kelkar, Shaunak ; Saleh, Bahram ; Yusa, Fumie ; Suzuki, Yohei ; Komatsu, Takafumi ; Webster, Thomas ( January 2021 , Orthopaedic Research Society)

   INTRODUCTION: Orthopedic implants are important therapeutic devices for the management of a wide range of orthopedic conditions. However, bacterial infections of orthopedic implants remain a major problem, and not an uncommon one, leading to an increased rate of osteomyelitis, sepsis, implant failure and dysfunction, etc. Treating these infections is more challenging as the causative organism protects itself by the production of a biofilm over the implant’s surface (1). Infections start by the adhesion and colonization of pathogenic bacteria such as Staphylococcus aureus (SA), Staphylococcus epidermidis (SE), Escherichia coli (E. coli), Methicillin-Resistant Staphylococcus aureus (MRSA), and Multi-Drug Resistant Escherichia coli (MDR E. coli) on the implant’s surfaces. Specifically, Staphylococcus comprises up to two-thirds of all pathogens involved in orthopedic implant infections (2). However, bacterial surface adhesion is a complex process influenced by several factors such as chemical composition, hydrophobicity, magnetization, surface charge, and surface roughness of the implant (3). Considering the intimate association between bacteria and the implant surface, we measured the effect of stainless-steel surface properties on bacterial surface attachment and subsequent formation of biofilms controlling above mentioned factors. METHODS: The prominent bacteria responsible for orthopedic implant infections (SA, SE, E. coli, MRSA, and MDR E. coli) were used inmore »this study. We were able to control the grain size of medical grade 304 and 316L stainless steel without altering their chemical composition (grain size range= 20μm-200nm) (4). Grain size control affected the nano-topography of the material surfaces which was measured by an Atomic Force Microscope (AFM). Grain sizes, such as 0.2, 0.5, 1, 2, 3, 9, and 10 μm, were used both polished and non-polished. All the stainless-steel samples were cleaned by treating with acetone and ethanol under sonication. Triplicates of all polished and non-polished samples with different grain sizes were subjected to magnetization of DM, 0.1T, 0.5T, and 1T, before seeding them with the bacteria. Controls were used in the form of untreated samples. Bacterial were grown in Tryptic Soy Broth (TSB). An actively growing bacterial suspension was seeded onto the stainless-steel discs into 24-well micro-titer plates and kept for incubation. After 24 hours of incubation, the stainless-steel discs were washed with Phosphate Buffer Saline (PBS) to remove the plankton bacteria and allow the sessile bacteria in the biofilm to remain. The degree of development of the bacterial biofilms on the stainless-steel discs were measured using spectrophotometric analysis. For this, the bacterial biofilm was removed from the stainless steel by sonication. The formation of biofilms was also determined by performing a biofilm staining method using Safranin. RESULTS SECTION: AFM results revealed a slight decrease in roughness by decreasing the grain size of the material. Moreover, the samples were segregated into two categories of polished and non-polished samples, in which polishing decreased roughness significantly. After careful analysis we found out that polished surfaces showed a higher degree for biofilm formation in comparison to the non-polished ones. We also observed that bacteria showed a higher rate for biofilm formation for the demagnetized samples, whereas 0.5T magnetization showed the least amount of biofilm formation. After 0.5T, there was no significant change in the rate of biofilm formation on the stainless-steel samples. Altogether, stainless steel samples containing 0.5 μm and less grainsize, and magnetized with 0.5 tesla and stronger magnets demonstrated the least degree of biofilm formation. DISCUSSION: In summary, the results demonstrate that controlling the grain size of medical grade stainless steel can control and mitigate bacterial responses on, and thus possibly infections of, orthopedic implants or other implantable devices. The research was funded by Komatsuseiki Kosakusho Co., Ltd (KSJ: Japan) SIGNIFICANCE/CLINICAL RELEVANCE: Orthopedic implants that more than 70% of them are made of metals (i.e., stainless steel, titanium, and cobalt-chromium alloys) are failing through loosening and breakage due to their limited mechanical properties. On the other hand, the risk of infection for these implants and its financial burden on our society is undeniable. We have seen that our uniformly nanograined stainless steel shows improved mechanical properties (i.e., higher stiffness, hardness, fatigue) as compared to conventional stainless steel along with the reduction of biofilm formation on its surface. These promising results made us to peruse the development of nanograined titanium and cobalt-chromium alloys for resolving the complications of orthopedic implants.« less
3. NanoSUS (Ultra Fine Grained Stainless-Steel) for Orthopedic Implants

   Bahram Saleh, Ada Vernet-Crua ( February 2021 , Orthopaedic Research Society)

   Statement of Purpose: Orthopedic implants are important therapeutic devices for the management of a wide range of orthopedic conditions. However, bacterial infections of orthopedic implants remain a major problem, and not an uncommon one, leading to an increased rate of osteomyelitis, sepsis, implant failure and dysfunction, etc. Treating these infections is more challenging as the causative organism protects itself by the production of a biofilm over the implant’s surface (1). Infections start by the adhesion and colonization of pathogenic bacteria such as Staphylococcus aureus (SA), Staphylococcus epidermidis (SE), Escherichia coli (E. coli), Methicillin-Resistant Staphylococcus aureus (MRSA), and Multi-Drug Resistant Escherichia coli (MDR E. coli) on the implant’s surfaces. Specifically, Staphylococcus comprises up to two-thirds of all pathogens involved in orthopedic implant infections (2). However, bacterial surface adhesion is a complex process influenced by several factors such as chemical composition, hydrophobicity, magnetization, surface charge, and surface roughness of the implant (3). Considering the intimate association between bacteria and the implant surface, we measured the effect of stainless-steel surface properties on bacterial surface attachment and subsequent formation of biofilms controlling above mentioned factors. Method: The prominent bacteria responsible for orthopedic implant infections (SA, SE, E. coli, MRSA, and MDR E. coli) weremore »used in this study. We were able to control the grain size of medical grade 304 and 316L stainless steel without altering their chemical composition (grain size range= 20μm-200nm) (4). Grain size control affected the nano-topography of the material surfaces which was measured by an Atomic Force Microscope (AFM). Grain sizes, such as 0.2, 0.5, 1, 2, 3, 9, and 10 μm, were used both polished and non-polished. All the stainless-steel samples were cleaned by treating with acetone and ethanol under sonication. Triplicates of all polished and non-polished samples with different grain sizes were subjected to magnetization of DM, 0.1T, 0.5T, and 1T, before seeding them with the bacteria. Controls were used in the form of untreated samples. Bacterial were grown in Tryptic Soy Broth (TSB). An actively growing bacterial suspension was seeded onto the stainless-steel discs into 24-well micro-titer plates and kept for incubation. After 24 hours of incubation, the stainless-steel discs were washed with Phosphate Buffer Saline (PBS) to remove the plankton bacteria and allow the sessile bacteria in the biofilm to remain. The degree of development of the bacterial biofilms on the stainless-steel discs were measured using spectrophotometric analysis. For this, the bacterial biofilm was removed from the stainless steel by sonication. The formation of biofilms was also determined by performing a biofilm staining method using Safranin. Results: AFM results revealed a slight decrease in roughness by decreasing the grain size of the material. Moreover, the samples were segregated into two categories of polished and non-polished samples, in which polishing decreased roughness significantly. After careful analysis we found out that polished surfaces showed a higher degree for biofilm formation in comparison to the non-polished ones. We also observed that bacteria showed a higher rate for biofilm formation for the demagnetized samples, whereas 0.5T magnetization showed the least amount of biofilm formation. After 0.5T, there was no significant change in the rate of biofilm formation on the stainless-steel samples. Altogether, stainless steel samples containing 0.5 μm and less grainsize, and magnetized with 0.5 tesla and stronger magnets demonstrated the least degree of biofilm formation. Conclusion: In summary, the results demonstrate that controlling the grain size of medical grade stainless steel can control and mitigate bacterial responses on, and thus possibly infections of, orthopedic implants or other implantable devices. The research was funded by Komatsuseiki Kosakusho Co., Ltd (KSJ: Japan)« less
4. Influence of substrate roughness on particle adhesion and concentration

   https://doi.org/10.1007/s42461-021-00521-9  

   Sansao, B.M.B. ( November 2021 , Mining metallurgy exploration)

   In this research study, the interaction of particles with substrates of different roughness magnitude was investigated. Particle surface treatment, relative humidity (RH), and surface roughness levels were controlled in order to achieve separation of different particles by applying removal forces. Three different approaches to reproducibly roughen surfaces were used. Initially, glass disks were laser engraved to create a reproducible, controlled roughness substrate. However, the laser engraving method produced surface features that were much greater in scale than the particles. These scale differences were such that the substrates produced were not of value to this research. The second option investigated to induce reproducible substrate roughness was to scratch the glass disk using sandpapers of known grain size. A third approach to establish reproducible roughness was to use fine stainless-steel wire mesh substrates. In tests with sanded glass disks, the interfacial energy of plasma-cleaned (hydrophilic) glass beads had a high variation at 40% RH, showing non-uniformity of area of contact between particles and substrates. As the RH increased, it was expected that the interfacial energy of hydrophilic particles would increase, but this behavior was not observed. In addition, comparing the interfacial energy results of hydrophilic particles with hydrophobic particles, a region withmore »significant interfacial energy difference was not identified. In the case of the stainless-steel mesh substrate, the mesh asperities and particle dimensions were comparable. Thus, the smaller particles had more area of contact with the substrate than the larger particles. For the plasma-cleaned (hydrophilic) beads, the recovery values had an average of 92.5% recovery when the RH was between 46 and 85%. For the hydrophobic beads, the average recovery was 19.0% when the RH was between 46 and 75%. Thus, the hydrophobic characteristic of the particle influenced its lower interaction with the mesh substrate. The difference in recovery can be exploited to achieve separation of particles based upon adhesive forces.« less
5. Orientation effects on the nanoscale adsorption behavior of bone morphogenetic protein-2 on hydrophilic silicon dioxide

   https://doi.org/10.1039/c8ra09165j  

   Marquetti, Izabele ; Desai, Salil ( January 2019 , RSC Advances)

   Bone Morphogenetic Protein-2 (BMP-2) is a growth factor associated with different developmental functions in regenerative medicine and tissue engineering. Because of its favorable properties for the development of bone and cartilage tissue, BMP-2 promotes the biocompatibility of medical implants. In this research, molecular dynamics simulations were implemented to simulate the interaction of BMP-2 with a flat hydrophilic silicon dioxide substrate, an important biomaterial for medical applications. We considered the influence of four orthogonal protein orientations on the adsorption behavior. Results showed that arginine and lysine were the main residues to interact with the silicon dioxide substrate, directly adsorbing onto the surface and overcoming water layers. However, between these charged residues, we observed a preference for arginine to adsorb. Orientations with the α-helix loop closer to the surface at the beginning of the simulations had greater loss of secondary structure as compared to the other configurations. Among all the orientations, the end-on B configuration had favorable adsorption characteristics with a binding energy of 14 000 kJ mol −1 and retention of 21.7% β-sheets as confirmed by the Ramachandran plots. This research provides new insights into the nanoscale interaction of BMP-2 and silicon dioxide substrate with applications in orthopedic implants and regenerative medicine.