Meniscus repair via collagen matrix wrapping and bone marrow injection: clinical and biomolecular study.

PURPOSE: The functional outcomes of arthroscopic matrix-based meniscus repair (AMMR) in patients two and five years after the treatment clearly show that the use of the collagen matrix and bone marrow aspirate creates favorable biological conditions for meniscus healing. This study not only provides ten follow-up results but also investigates biomolecular mechanisms governing the regenerative process. METHODS: Case series was based on data collected from patients who underwent AMMR procedure, starting with preoperatively through two-year and five-year till ten-year follow-up. The outcomes are presented as IKDC and the Lysholm subjective scores as well as the imaging results. Biomolecular investigation of the membranes utilized in the AMMR procedure include DNA content analysis, cell viability and proliferation study of bone marrow and bone marrow concentrate-derived cells, and cytokine array performed on monocytes cultured on the membranes. CONCLUSION: Data collected from patients who underwent AMMR procedure, starting with pre-operatively through two year and five year till ten year follow-up, indicate the possibility for long-term, stable meniscus preservation. Outcomes are manifested with a visible improvement of the IKDC and the Lysholm subjective scores as well as in the imaging results. The type of the meniscal tear or complexity of the knee injury (isolated AMMR vs. AMMR + ACL) did not affect the clinical outcomes. The study highlighted the role of the membrane in facilitating cell adhesion and proliferation. Additionally, several cytokines were selected as potentially crucial products of the membrane vs. monocyte interactions, driving the tissue regeneration and remodeling. Interestingly, thresholds of what constitutes a safe and well-decellularized membrane according to relevant literature have been significantly breached, but ultimately did not elicit detrimental side effects.

Radiosensitizing properties of magnetic hyperthermia mediated by superparamagnetic iron oxide nanoparticles (SPIONs) on human cutaneous melanoma cell lines.

Melanoma is responsible for the majority of deaths related to skin cancer. Worryingly, prognoses show an increasing number of melanoma cases each year worldwide. Radiotherapy, which is a cornerstone of cancer treatment, has proved to be useful but insufficient in melanoma management due to exceptionally high radioresistance of melanoma cells. This problem could be overcome by superparamagnetic iron oxide nanoparticles (SPIONs) used as heat mediators in magnetic hyperthermia, which not only enhance radiosensitivity, but also enable precise targeting by exploitation of their magnetic properties.

Correlations between serum adipocytokine concentrations, disease stage, radiological status and total body fat content in the patients with primary knee osteoarthritis.

PURPOSE: The study was designed to investigate whether serum concentrations of leptin, resistin and adiponectin in obese and normal-weight patients with primary knee osteoarthritis (OA) correlate with clinical and radiological stages of the disease and percentage of total body fat. METHODS: Seventy-three patients with knee OA, divided into obese and normal-weight groups, were clinically evaluated according to the Knee Society Score (KSS), and radiologically assessed using Kellgren and Lawrence scale. The percentage of total body fat and some anthropometric data were also given. Serum leptin, resistin and adiponectin concentrations were measured by Elisa and were correlated with the clinical, radiological and anthropometric parameters. RESULTS: Leptin concentrations were significantly higher (p = 0.001) in the obese patients and positively correlated (R = 0.63) with radiologically assessed OA grade, but only in the normal-weight group. Resistin and adiponectin concentrations were identical in obese and normal-weight patients and negatively correlated (R = -0.41) with the clinical status of obese patients. In both groups, percentage of total body fat positively correlated (R = 0.29 and R = 0.53 for obese and normal-weight respectively) with radiologically assessed OA grade. However, no correlations were found with clinical status of the patients. CONCLUSIONS: It was found that in the obese patients with knee OA, increased percentage of total body fat and elevated serum leptin concentration might favour the advancement of clinical but not radiologically assessed changes in the joint structures, while in normal-weight patients it correlates only with radiologically assessed changes but does not affect to an appreciable extent the clinical status of the patients.

Formulation and evaluation of a bioink composed of alginate, gelatin, and nanocellulose for meniscal tissue engineering.

1The necessity to preserve meniscal function prompts the research and development of novel treatment options, like three-dimensional (3D) bioprinting. However, bioinks for meniscal 3D bioprinting have not been extensively explored. Therefore, in this study, a bioink composed of alginate, gelatin, and carboxymethylated cellulose nanocrystal (CCNC) was formulated and evaluated. Firstly, bioinks with varying concentrations of the aforementioned components were subjected to rheological analysis (amplitude sweep test, temperature sweep test, and rotation). The optimal bioink formulation of 4.0% gelatin, 0.75% alginate, and 1.4% CCNC dissolved in 4.6% D-mannitol was further used for printing accuracy analysis, followed by 3D bioprinting with normal human knee articular chondrocytes (NHAC-kn). The encapsulated cells' viability was > 98%, and collagen II expression was stimulated by the bioink. The formulated bioink is printable, stable under cell culture conditions, biocompatible, and able to maintain the native phenotype of chondrocytes. Aside from meniscal tissue bioprinting, it is believed that this bioink could serve as a basis for the development of bioinks for various tissues.

Hyaluronic acid and multiwalled carbon nanotubes as bioink additives for cartilage tissue engineering.

Articular cartilage and meniscus injuries are prevalent disorders with insufficient regeneration responses offered by available treatment methods. In this regard, 3D bioprinting has emerged as one of the most promising new technologies, offering novel treatment options. Additionally, the latest achievements from the fields of biomaterials and tissue engineering research identified constituents facilitating the creation of biocompatible scaffolds. In this study, we looked closer at hyaluronic acid and multi-walled carbon nanotubes as bioink additives. Firstly, we assessed the minimal concentrations that stimulate cell viability, and decrease reactive oxygen species and apoptosis levels in 2D cell cultures of normal human knee articular chondrocytes (NHAC) and human adipose-derived mesenchymal stem cells (hMSC-AT). In this regard, 0.25 mg/ml of hyaluronic acid and 0.0625 mg/ml of carbon nanotubes were selected as the most optimal concentrations. In addition, we investigated the protective influence of 2-phospho-L-ascorbic acid in samples with carbon nanotubes. Tests conducted on 3D bioprinted constructs revealed that only a combination of components positively impacted cell viability throughout the whole experiment. Gene expression analysis of COL1A1, COL6A1, HIF1A, COMP, RUNX2, and POU5F1 showed significant changes in the expression of all analyzed genes with a progressive overall loss of transcriptional activity in most of them.

A cohort study reveals different dynamics of SARS-CoV-2-specific antibody formation after Comirnaty and Vaxzevria vaccination.

The Coronavirus (COVID-19) Disease Pandemic, caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has affected millions of people worldwide, prompting a collective effort from the global scientific community to develop a vaccine against it. This study purports to investigate the influence of factors such as sex, age, type of vaccination (Comirnaty, BNT162b2, Pfizer Inc. or Vaxzevria, ChAdOx1-S, Oxford/AstraZeneca), and time since vaccine administration on the process of antibody production. Both of them are based on the introduction of SARS-CoV-2 spike protein (S protein) to the body using different mechanisms (mRNA and recombinant adenovirus, respectively). S protein is responsible for host cell attachment and penetration via its receptor-binding domain (RBD domain). The level of anti-RBD IgG antibodies was tested with an ELISA-based immunodiagnostic assay in serum samples from a total of 1395 patients at 3 time points: before vaccination, after the first dose, and after the second dose. Our novel statistical model, the Generalized Additive Model, revealed variability in antibody production dynamics for both vaccines. Interestingly, no discernible variation in antibody levels between men and women was found. A nonlinear relationship between age and antibody production was observed, characterized by decreased antibody levels for people up to 30 and over 60 years of age, with a lack of correlation in the middle age range. Collectively, our findings further the understanding of the mechanism driving vaccine-induced immunity. Additionally, we propose the Generalized Additive Model as a standardized way of presenting data in similar research.

3D Bioprinting in Skin Related Research: Recent Achievements and Application Perspectives.

In recent years, significant progress has been observed in the field of skin bioprinting, which has a huge potential to revolutionize the way of treatment in injury and surgery. Furthermore, it may be considered as an appropriate platform to perform the assessment and screening of cosmetic and pharmaceutical formulations. Therefore, the objective of this paper was to review the latest advances in 3D bioprinting dedicated to skin applications. In order to explain the boundaries of this technology, the architecture and functions of the native skin were briefly described. The principles of bioprinting methods were outlined along with a detailed description of key elements that are required to fabricate the skin equivalents. Next, the overview of recent progress in 3D bioprinting studies was presented. The article also highlighted the potential applications of bioengineered skin substituents in various fields including regenerative medicine, modeling of diseases, and cosmetics/drugs testing. The advantages, limitations, and future directions of this technology were also discussed.

Photodegradation of Biohazardous Dye Brilliant Blue R Using Organometallic Silver Nanoparticles Synthesized through a Green Chemistry Method.

Nowadays, nanostructures having tremendous chemical and physical properties are gaining attention in the biomedical industry. However, when they are prepared through classical methods (physical and chemical), they are often non-biocompatible and toxic. Considering the mentioned factors, in this research, organometallic silver nanostructures (OMAgNs) have been prepared by the green chemistry method using the acetone, methanol, and methanol-hexane-based extracts of the medicinally important plant Cichorium intybus. Secondary metabolites from C. intybus can be used as an alternative to synthetic reagents at an industrial scale to manufacture biosafe and economical nanostructures with enhanced physicochemical parameters. Prepared nanostructures were characterized using SEM, XRD, FTIR, TGA, UV, and zeta potential measurement. SEM analysis revealed different shapes of OMAgNs, prepared with various extracts. XRD analysis showed the crystallinity of the nanostructures. FTIR spectroscopy helped to identify groups of compounds present in the extracts and used for the OMAgNs synthesis. Out of the three tested OMAgNs, those prepared with methanol extract were selected due to the highest obtained yield and stability (highest negative zeta potential) and were tested as a cost-efficient and active agent to photodegrade organic pollutant, Brilliant Blue R, using energy from sunlight. A decrease in UV-VIS absorbance confirmed the rapid degradation of the dye.

Bioevaluation of superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with dihexadecyl phosphate (DHP).

Superparamagnetic iron oxide nanoparticles (SPIONs) have been investigated for wide variety of applications. Their unique properties render them highly applicable as MRI contrast agents, in magnetic hyperthermia or targeted drug delivery. SPIONs surface properties affect a whole array of parameters such as: solubility, toxicity, stability, biodistribution etc. Therefore, progress in the field of SPIONs surface functionalization is crucial for further development of therapeutic or diagnostic agents. In this study, SPIONs were synthesized by thermal decomposition of iron (III) acetylacetonate Fe(acac)3 and functionalized with dihexadecyl phosphate (DHP) via phase transfer. Bioactivity of the SPION-DHP was assessed on SW1353 and TCam-2 cancer derived cell lines. The following test were conducted: cytotoxicity and proliferation assay, reactive oxygen species (ROS) assay, SPIONs uptake (via Iron Staining and ICP-MS), expression analysis of the following genes: alkaline phosphatase (ALPL); ferritin light chain (FTL); serine/threonine protein phosphatase 2A (PP2A); protein tyrosine phosphatase non-receptor type 11 (PTPN11); transferrin receptor 1 (TFRC) via RT-qPCR. SPION-DHP nanoparticles were successfully obtained and did not reveal significant cytotoxicity in the range of tested concentrations. ROS generation was elevated, however not correlated with the concentrations. Gene expression profile was slightly altered only in SW1353 cells.

Utilization of Carbon Nanotubes in Manufacturing of 3D Cartilage and Bone Scaffolds.

Cartilage and bone injuries are prevalent ailments, affecting the quality of life of injured patients. Current methods of treatment are often imperfect and pose the risk of complications in the long term. Therefore, tissue engineering is a rapidly developing branch of science, which aims at discovering effective ways of replacing or repairing damaged tissues with the use of scaffolds. However, both cartilage and bone owe their exceptional mechanical properties to their complex ultrastructure, which is very difficult to reproduce artificially. To address this issue, nanotechnology was employed. One of the most promising nanomaterials in this respect is carbon nanotubes, due to their exceptional physico-chemical properties, which are similar to collagens-the main component of the extracellular matrix of these tissues. This review covers the important aspects of 3D scaffold development and sums up the existing research tackling the challenges of scaffold design. Moreover, carbon nanotubes-reinforced bone and cartilage scaffolds manufactured using the 3D bioprinting technique will be discussed as a novel tool that could facilitate the achievement of more biomimetic structures.

Osteoarthritis Severely Decreases the Elasticity and Hardness of Knee Joint Cartilage: A Nanoindentation Study.

The nanoindentation method was applied to determine the elastic modulus and hardness of knee articular cartilage. Cartilage samples from both high weight bearing (HWB) and low weight bearing (LWB) femoral condyles were collected from patients diagnosed with osteoarthritis (OA). The mean elastic modulus of HWB cartilage was 4.46 ± 4.44 MPa in comparison to that of the LWB region (9.81 ± 8.88 MPa, p < 0.001). Similarly, the hardness was significantly lower in HWB tissue (0.317 ± 0.397 MPa) than in LWB cartilage (0.455 ± 0.434 MPa, p < 0.001). When adjusted to patients' ages, the mean elastic modulus and hardness were both significantly lower in the age group over 70 years (p < 0.001). A statistically significant difference in mechanical parameters was also found in grade 3 and 4 OA. This study provides an insight into the nanomechanical properties of the knee articular cartilage and provides a starting point for personalized cartilage grafts that are compatible with the mechanical properties of the native tissue.

Assembly and Characterization of HBc Derived Virus-like Particles with Magnetic Core.

Core-virus like particles (VLPs) assembly is a kinetically complex cascade of interactions between viral proteins, nanoparticle's surface and an ionic environment. Despite many in silico simulations regarding this process, there is still a lack of experimental data. The main goal of this study was to investigate the capsid protein of hepatitis B virus (HBc) assembly into virus-like particles with superparamagnetic iron oxide nanoparticles (SPIONs) as a magnetic core in relation to their characteristics. The native form of HBc was obtained via agroinfection of Nicotiana benthamiana with pEAQ-HBc plasmid. SPIONs of diameter of 15 nm were synthesized and functionalized with two ligands, providing variety in ζ-potential and hydrodynamic diameter. The antigenic potential of the assembled core-VLPs was assessed with enzyme-linked immunosorbent assay (ELISA). Morphology of SPIONs and core-VLPs was evaluated via transmission electron microscopy (TEM). The most successful core-VLPs assembly was obtained for SPIONs functionalized with dihexadecyl phosphate (DHP) at SPIONs/HBc ratio of 0.2/0.05 mg/mL. ELISA results indicate significant decrease of antigenicity concomitant with core-VLPs assembly. In summary, this study provides an experimental assessment of the crucial parameters guiding SPION-HBc VLPs assembly and evaluates the antigenicity of the obtained structures.

Composite spheres made of bioengineered spider silk and iron oxide nanoparticles for theranostics applications.

Bioengineered spider silk is a biomaterial that has exquisite mechanical properties, biocompatibility, and biodegradability. Iron oxide nanoparticles can be applied for the detection and analysis of biomolecules, target drug delivery, as MRI contrast agents and as therapeutic agents for hyperthermia-based cancer treatments. In this study, we investigated three bioengineered silks, MS1, MS2 and EMS2, and their potential to form a composite material with magnetic iron oxide nanoparticles (IONPs). The presence of IONPs did not impede the self-assembly properties of MS1, MS2, and EMS2 silks, and spheres formed. The EMS2 spheres had the highest content of IONPs, and the presence of magnetite IONPs in these carriers was confirmed by several methods such as SEM, EDXS, SQUID, MIP-OES and zeta potential measurement. The interaction of EMS2 and IONPs did not modify the superparamagnetic properties of the IONPs, but it influenced the secondary structure of the spheres. The composite particles exhibited a more than two-fold higher loading efficiency for doxorubicin than the plain EMS2 spheres. For both the EMS2 and EMS2/IONP spheres, the drug revealed a pH-dependent release profile with advantageous kinetics for carriers made of the composite material. The composite spheres can be potentially applied for a combined cancer treatment via hyperthermia and drug delivery.

Parenteral-Oral Immunization with Plant-Derived HBcAg as a Potential Therapeutic Vaccine against Chronic Hepatitis B.

Chronic hepatitis B (CHB) is the cause of severe liver damage, cirrhosis, and hepatocellular carcinoma for over 240 million people worldwide. Nowadays, several types of treatment are being investigated, including immunotherapy using hepatitis B core antigen (HBcAg) assembled into highly immunogenic capsid-like particles (CLPs). Immunogenicity of plant-produced and purified HBcAg, administered parenterally or intranasally, was previously reported. In this study, a novel parenteral-oral vaccination scheme is proposed using plant-derived HBcAg preparations. The antigen for injection was obtained via transient expression in Nicotiana benthamiana. HBcAg-producing transgenic lettuce was lyophilized and used as an orally delivered booster. The intracellular location of plant-produced HBcAg CLPs implies additional protection in the digestive tract during oral immunization. BALB/c mice were intramuscularly primed with 10 µg of the purified antigen and orally boosted twice with 5 or 200 ng of HBcAg. A long-lasting and significant systemic response after boosting with 200 ng HBcAg was induced, with anti-HBc titer of 25,000. Concomitantly, an insignificant mucosal response was observed, with an S-IgA titer of only 500. The profile of IgG isotypes indicates a predominant Th1 type of immune response, supplemented by Th2, after injection-oral vaccination. The results demonstrate that a low dose of parenteral-oral immunization with plant-derived HBcAg can elicit a specific and efficient response. This study presents a potential new pathway of CHB treatment.

BMV Propagation, Extraction and Purification Using Chromatographic Methods.

Brome mosaic virus (BMV) is a well-known plant virus representing single-stranded RNA (ssRNA) positive-sense viruses. It has been widely used as a model in multiple studies concerning plant virus biology, epidemiology and the application of viral capsids in nanotechnology. Herein, we describe a method for BMV purification based on ion-exchange and size-exclusion chromatography. The presented method is of similar efficiency to previously described protocols relying on differential centrifugation and can easily be scaled up. The resulting BMV capsids are stable and monodisperse and can be used for further applications.

Biophysical analysis of BMV virions purified using a novel method.

Brome mosaic virus (BMV) has been successfully loaded with different types of nanoparticles. However, studies concerning its application as a nanoparticle carrier demand high-purity virions in large amounts. Existing BMV purification protocols rely on multiple differential ultracentrifugation runs of the initially purified viral preparation. Herein, we describe an alternative method for BMV purification based on ion-exchange chromatography and size-exclusion chromatography (SEC) yielding 0.2mg of virus from 1g of plant tissue. Our method is of similar efficiency to previously described protocols and can easily be scaled up. The method results in high-quality BMV preparations as confirmed by biophysical analyses, including cryogenic transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS), static light scattering (SLS), and circular dichroism (CD) measurements and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy. Our results revealed that purified BMV capsids are stable and monodisperse and can be used for further downstream applications. In this work, we also characterize secondary structure and size fluctuations of the BMV virion at different pH values.