Decellularization and characterization of camel pericardium as a new scaffold for tissue engineering and regenerative medicine.

BACKGROUND: Valvular heart diseases (VHDs) have become prevalent in populations due to aging. Application of different biomaterials for cardiac valve regeneration and repair holds a great promise for treatment of VHD. Aortic valve replacement using tissue-engineered xenografts is a considered approach, and the pericardium of different species such as porcine and bovine has been studied over the last few years. It has been suggested that the animal origin can affect the outcomes of replacement. METHODS: So, herein, we at first decellularized and characterized the camel pericardium (dCP), then characterized dCP with H&E staining, in vitro and in vivo biocompatibility and mechanical tests and compared it with decellularized bovine pericardium (dBP), to describe the potency of dCP as a new xenograft and bio scaffold. RESULTS: The histological assays indicated less decluttering and extracellular matrix damage in dCP after decellularization compared to the dBP also dCP had higher Young Modulus (105.11), and yield stress (1.57 ± 0.45). We observed more blood vessels and also less inflammatory cells in the dCP sections after implantation. CONCLUSIONS: In conclusion, the results of this study showed that the dCP has good capabilities not only for use in VHD treatment but also for other applications in tissue engineering and regenerative medicine.

Potentials and future perspectives of multi-target drugs in cancer treatment: the next generation anti-cancer agents.

Cancer is a major public health problem worldwide with more than an estimated 19.3 million new cases in 2020. The occurrence rises dramatically with age, and the overall risk accumulation is combined with the tendency for cellular repair mechanisms to be less effective in older individuals. Conventional cancer treatments, such as radiotherapy, surgery, and chemotherapy, have been used for decades to combat cancer. However, the emergence of novel fields of cancer research has led to the exploration of innovative treatment approaches focused on immunotherapy, epigenetic therapy, targeted therapy, multi-omics, and also multi-target therapy. The hypothesis was based on that drugs designed to act against individual targets cannot usually battle multigenic diseases like cancer. Multi-target therapies, either in combination or sequential order, have been recommended to combat acquired and intrinsic resistance to anti-cancer treatments. Several studies focused on multi-targeting treatments due to their advantages include; overcoming clonal heterogeneity, lower risk of multi-drug resistance (MDR), decreased drug toxicity, and thereby lower side effects. In this study, we'll discuss about multi-target drugs, their benefits in improving cancer treatments, and recent advances in the field of multi-targeted drugs. Also, we will study the research that performed clinical trials using multi-target therapeutic agents for cancer treatment.

Microfluidic Sensor Based on Cell-Imprinted Polymer-Coated Microwires for Conductometric Detection of Bacteria in Water.

The rapid, inexpensive, and on-site detection of bacterial contaminants using highly sensitive and specific microfluidic sensors is attracting substantial attention in water quality monitoring applications. Cell-imprinted polymers (CIPs) have emerged as robust, cost-effective, and versatile recognition materials with selective binding sites for capturing whole bacteria. However, electrochemical transduction of the binding event to a measurable signal within a microfluidic device to develop easy-to-use, compact, portable, durable, and affordable sensors remains a challenge. For this paper, we employed CIP-functionalized microwires (CIP-MWs) with an affinity towards E. coli and integrated them into a low-cost microfluidic sensor to measure the conductometric transduction of CIP-bacteria binding events. The sensor comprised two CIP-MWs suspended perpendicularly to a PDMS microchannel. The inter-wire electrical resistance of the microchannel was measured before, during, and after exposure of CIP-MWs to bacteria. A decline in the inter-wire resistance of the sensor after 30 min of incubation with bacteria was detected. Resistance change normalization and the subsequent analysis of the sensor's dose-response curve between 0 to 109 CFU/mL bacteria revealed the limits of detection and quantification of 2.1 × 105 CFU/mL and 7.3 × 105 CFU/mL, respectively. The dynamic range of the sensor was 104 to 107 CFU/mL where the bacteria counts were statistically distinguishable from each other. A linear fit in this range resulted in a sensitivity of 7.35 µS per CFU/mL. Experiments using competing Sarcina or Listeria cells showed specificity of the sensor towards the imprinted E. coli cells. The reported CIP-MW-based conductometric microfluidic sensor can provide a cost-effective, durable, portable, and real-time solution for the detection of pathogens in water.

Nanopore sensors for viral particle quantification: current progress and future prospects.

Rapid, inexpensive, and laboratory-free diagnostic of viral pathogens is highly critical in controlling viral pandemics. In recent years, nanopore-based sensors have been employed to detect, identify, and classify virus particles. By tracing ionic current containing target molecules across nano-scale pores, nanopore sensors can recognize the target molecules at the single-molecule level. In the case of viruses, they enable discrimination of individual viruses and obtaining important information on the physical and chemical properties of viral particles. Despite classical benchtop virus detection methods, such as amplification techniques (e.g., PCR) or immunological assays (e.g., ELISA), that are mainly laboratory-based, expensive and time-consuming, nanopore-based sensing methods can enable low-cost and real-time point-of-care (PoC) and point-of-need (PoN) monitoring of target viruses. This review discusses the limitations of classical virus detection methods in PoN virus monitoring and then provides a comprehensive overview of nanopore sensing technology and its emerging applications in quantifying virus particles and classifying virus sub-types. Afterward, it discusses the recent progress in the field of nanopore sensing, including integrating nanopore sensors with microfabrication technology, microfluidics and artificial intelligence, which have been demonstrated to be promising in developing the next generation of low-cost and portable biosensors for the sensitive recognition of viruses and emerging pathogens.

Conventional and microfluidic methods for airborne virus isolation and detection.

With the COVID-19 pandemic, the threat of infectious diseases to public health and safety has become much more apparent. Viral, bacterial and fungal diseases have led to the loss of millions of lives, especially in the developing world. Diseases caused by airborne viruses like SARS-CoV-2 are difficult to control, as these viruses are easily transmissible and can circulate in the air for hours. To contain outbreaks of viruses such as SARS-CoV-2 and institute targeted precautions, it is important to detect them in air and understand how they infect their targets. Point-of-care (PoC) diagnostics and point-of-need (PoN) detection methods are necessary to rapidly test patient and environmental samples, so precautions can immediately be applied. Traditional benchtop detection methods such as ELISA, PCR and culture are not suitable for PoC and PoN monitoring, because they can take hours to days and require specialized equipment. Microfluidic devices can be made at low cost to perform such assays rapidly and at the PoN. They can also be integrated with air- and liquid-based sampling technologies to capture and analyze viruses from air and body fluids. Here, conventional and microfluidic virus detection methods are reviewed and compared. The use of air sampling devices to capture and concentrate viruses is discussed first, followed by a review of analysis methods such as immunoassays, RT-PCR and isothermal amplification in conventional and microfluidic platforms. This review provides an overview of the capabilities of microfluidics in virus handling and detection, which will be useful to infectious disease researchers, biomedical engineers, and public health agencies.

Integration of microfluidic sample preparation with PCR detection to investigate the effects of simultaneous DNA-Inhibitor separation and DNA solution exchange.

In this paper, we applied a curved-channel microfluidic device to separate DNA from PCR-inhibitor-containing water and simultaneously wash them into clean water for detection using a portable PCR thermocycler. Environmental DNA (eDNA) sampling has become an effective surveying approach for detecting rare organisms. However, low concentration eDNA molecules may be masked by PCR inhibitors during amplification and detection, increasing the risk of false negatives. Therefore, technologies for on-site DNA separation and washing are urgently needed. Our device consisted of a half-circle microchannel with a DNA-inhibitor sample inlet, a clean buffer inlet, and multiple outlets. By using the flow-induced inertial forces, 10 µm DNA-conjugated microparticles were focused at the inner-wall of the curved microchannel while separation from 1 µm inhibitor-conjugated microparticles and DNA washing were achieved simultaneously with the Dean flow. We achieved singleplex focusing, isolation and washing of 10 µm particles at an efficiency of 94.5 ± 2.0%. In duplex experiments with 1 µm and 10 µm particles, larger particles were washed with an efficiency of 92.1 ± 1.6% and a purity of 79 ± 2%. By surface-functionalizing the microparticles with affinity groups against Atlantic salmon DNA and humic acid (HA), and processing samples of various concentrations in our device, we achieved an effective purification and detection of DNA molecules using the portable PCR thermocycler. Our method significantly decreased PCR quantitation cycles from Cq > 38 to Cq = 30.35 ± 0.5, which confirmed enhancement of PCR amplification. The proposed device takes a promising step forward in sample preparation towards an integrated device that can be used for simultaneous purification and solution exchange of DNA in point-of-need environmental monitoring applications.

The influence of polycaporolacton fumarate coating on mechanical properties and in vitro behavior of porous diopside-hardystonite nano-composite scaffold.

One of the significant challenges in bone tissue engineering is the fabrication of highly porous scaffolds with interconnected pores and appropriate mechanical properties. Artificial scaffolds which used in the field of medicine are usually made of single phase of polymer or ceramic. However, composition of these materials can produce the scaffolds with improve mechanical and biological properties.The aim of this study is to synthesize three-dimensional hardystonite-diopside (HT-Dio) porous scaffolds modified by polycaporolacton fumarate coating for low-load-bearing bone tissue engineering applications. The results showed that hardystonite scaffolds with 15 wt. % diopside and 6 w/v % polymer polycaporolacton fumarate (PCLF) had a significant bioactivity. The cell culture and cell attachment assay results revealed the well spreading of BMS cells on the surface of modified scaffolds which indicates the high biocompatibility of this scaffold. The modified scaffolds had a mean pore size, porosity, compressive strength, modules and toughness of 293.47 ±â€¯5.51 µm, 74% ±â€¯1.01, 3.37 ±â€¯0.6 MPa, 151 ±â€¯1.1 MPa and 31.3 ±â€¯0.32 kJ/m3, respectively, which are in the appropriate range for spongy bone and hence can be a good candidate for bone tissue engineering applications.

In vitro evaluation of diopside/baghdadite bioceramic scaffolds modified by polycaprolactone fumarate polymer coating.

Porous Si-based ceramic scaffolds are widely attracted in biomedical tissue engineering application. Despite the attractive properties of these materials, their weak mechanical properties and high degradability in vitro and in vivo environment can limit their application as biomedical devises. Applying a thin layer of polymer on the surface of porous scaffolds can improve the mechanical properties and control the degradation rate. In this study, we produced new modified scaffolds with polymers coating in order to improved mechanical and biological properties of Si-based ceramics scaffolds. The results showed that applying 6 wt% PCLF polymer on the surface of Bagh-15 wt%Dio scaffolds delayed apatite formation compared to unmodified scaffolds. On the other hand, in the modified scaffolds, apatite formation was observed. The degradation rate of unmodified scaffolds was decreased around 82% after 28 days soaking in PBS solution. Based on the MTT assay and SEM micrographs, the BMS cells were spread and attached well on the surface of the scaffolds, which indicated a good biocompatibility. The results showed that these scaffolds have the potential to be used as a temporary substrate for bone tissue engineering application.

The combined effects of three-dimensional cell culture and natural tissue extract on neural differentiation of P19 embryonal carcinoma stem cells.

Tissue engineering, as a novel transplantation therapy, aims to create biomaterial scaffolds resembling the extracellular matrix in order to regenerate the damaged tissues. Adding bioactive factors to the scaffold would improve cell-tissue interactions. In this study, the effect of chitosan polyvinyl alcohol nanofibres containing carbon nanotube scaffold with or without active bioglass (BG+ /BG- ), in combination with neonatal rat brain extract on cell viability, proliferation, and neural differentiation of P19 embryonic carcinoma stem cells was investigated. To induce differentiation, the cells were cultured in α-MEM supplemented with neonatal rat brain extract on the scaffolds. The expression of undifferentiated stem cell markers as well as neuroepithelial and neural-specific markers was evaluated and confirmed by real-time Reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence procedures. Finally, the three-dimensional (3D) cultured cells were implanted into the damaged neural tubes of chick embryos, and their fates were followed in ovo. Based on the histological and immunofluorescence observations, the transplanted cells were able to survive, migrate, and penetrate into the host embryonic tissues. Gene network analysis suggested the possible involvement of neurotransmitters as a downstream target of synaptophysin and tyrosine hydroxylase. Overall, the results of this study indicated that combining the effects of 3D cell culture and natural brain tissue extract can accelerate the differentiation of P19 embryonic carcinoma cells into neuronal phenotype cells.

Hardystonite-Coated Poly(l-lactide) Nanofibrous Scaffold and Efficient Osteogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells.

In this study, a ceramic-coated nanofibrous scaffold has been fabricated to biomimic the microstructure of natural extracellular matrix and the stiffening inorganic compartment of bone. Poly-l-lactic acid (PLLA) nanofibers were electrospun and exposed to oxygen plasma to induce hydrophilicity and promote ceramic adsorption. Hardystonite (HS), which possesses superior osteoinduction potential over hydroxyapatite, was coated on plasma-treated PLLA nanofibers by drenching the nanofibers in HS suspension. Pure and composite PLLA-based scaffolds were characterized in terms of physical and biological properties. In vitro cultivation of adipose-derived mesenchymal stem cells (AMSCs) on the scaffolds displayed that the composite scaffold is able to further support cell attachment and proliferation. In case of osteogenic differentiation of AMSCs, HS coating significantly increased the synthesis and activity of alkaline phosphate over 21 days period. In addition, the composite scaffold showed improved mineralization. The expression level of osteonectin and osteocalcin genes was significantly enhanced by HS coating of nanofibers. The biological improvement of PLLA nanofibrous matrix in the presence of HS nanoparticles could either be attributed to the release and stimulatory effect of constituent ions of HS or to the modification of chemico-physical properties of the resultant ceramic by silicon and zinc present in HS.

Investigation of Osteoinductive Effects of Different Compositions of Bioactive Glass Nanoparticles for Bone Tissue Engineering.

Bioactive glasses (BG) is one of the well-known materials that used as dental and bone implants, for this reason it is always interesting for researchers has been to increase BG efficiency in the bone tissue engineering. The aim of this study was to evaluate the osteoinductivity of BG different composition nanoparticles with SiO2-CaO-P2O5. The 45S, 58S, and 63S compositions were prepared via the sol-gel technique. Characterization techniques such as x-ray diffraction, field emission scanning electron microscopy (FE-SEM), and laser Doppler electrophoresis (LDE) were used. The osteoinductive capacity of prepared nanoparticles was investigated using unrestricted somatic stem cells (USSC). The particle size of the samples with an amorphous structure mainly ranged less than 40 nm. The zeta potential was negative for all compositions in distilled water at pH 7.4. Bioactive glass nanoparticles were shown to support proliferation of USSC, as shown by microculture tetrazolium (MTT) assay. During osteogenic differentiation, significantly highest values of alkaline phosphatase (ALP) activity and biomineralization were observed on 45S BG. Subsequently, these markers were measured in higher amounts in USSC on 58S and 63S BG compared with tissue culture polystyrene. The nanometric particle size, osteoinductivity, and negative zeta potential make this material a possible candidate for bone tissue engineering applications.

Coating of electrospun poly(lactic-co-glycolic acid) nanofibers with willemite bioceramic: improvement of bone reconstruction in rat model.

We have investigated the combination effects of bioceramics and poly(lactide-co-glycolide) (PLGA) on bone reconstruction in calvarial critical size defects using a rat model. Willemite (Zn2SiO4) ceramics were prepared and coated on the surface of electrospun fabricated scaffolds. After scaffolds and nanoparticles characterization, osteoconductivity of the construct was analyzed using digital mammography, multislice spiral-computed tomography (MSCT) imaging, and histological analysis. Eight weeks after implantation, no sign of inflammation was observed at the site of the osseous defect. The results showed that the ceramics supported bone regeneration and highest bone reconstruction were observed in willemite-coated PLGA. This suggests that electrospun PLGA nanofibers coated with BG are potential candidate implants for bone tissue engineering applications.

Fluoride release and bioactivity evaluation of glass ionomer: Forsterite nanocomposite.

BACKGROUND: The most important limitation of glass ionomer cements (GICs) is the weak mechanical properties. Our previous research showed that higher mechanical properties could be achieved by addition of forsterite (Mg2SiO4) nanoparticles to ceramic part of GIC. The objective of the present study was to fabricate a glass ionomer- Mg2SiO4 nanocomposite and to evaluate the effect of addition of Mg2SiO4 nanoparticles on bioactivity and fluoride release behavior of prepared nanocomposite. MATERIALS AND METHODS: Forsterite nanoparticles were made by sol-gel process. X-ray diffraction (XRD) technique was used in order to phase structure characterization and determination of grain size of Mg2SiO4 nanopowder. Nanocomposite was fabricated via adding 3wt.% of Mg2SiO4 nanoparticles to ceramic part of commercial GIC (Fuji II GC). Fluoride ion release and bioactivity of nanocomposite were measured using the artificial saliva and simulated body fluid (SBF), respectively. Bioactivity of specimens was investigated by Fourier transitioned-infrared spectroscopy (FTIR), scanning electronmicroscopy (SEM), Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) and registration of the changes in pH of soaking solution at the soaking period. Statistical analysis was carried out by one Way analysis of variance and differences were considered significant if P < 0.05. RESULTS: The results of XRD analysis confirmed that nanocrystalline and pure Mg2SiO4 powder was obtained. Fluoride ion release evaluation showed that the values of released fluoride ions from nanocomposite are somewhat less than Fuji II GC. SEM images, pH changes of the SBF and results of the ICP-OES and FTIR tests confirmed the bioactivity of the nanocomposite. Statistical analysis showed that the differences between the results of all groups were significant (P < 0.05). CONCLUSION: Glass ionomer- Mg2SiO4 nanocomposite could be a good candidate for dentistry and orthopedic applications, through of desirable fluoride ion release and bioactivity.

Genotoxicity effects of nano bioactive glass and Novabone bioglass on gingival fibroblasts using single cell gel electrophoresis (comet assay): An in vitro study.

BACKGROUND: The greater surface of bioactive glass nanoparticles presents an incomparable and promising feature similar to the biological apatite. Nanoparticles improve cellular adhesion, enhance osteoblast proliferation and differentiation, and increase biomineralization for periodontal regeneration and dental implants. Considering the fact that interaction between periodontal cells and bone graft materials are important for periodontal lesion regeneration, the present study was undertaken to investigate the genotoxicity of a novel synthesized nanoscale bioactive glass and compared it with Novabone bioglass in periodontal fibroblasts cells, in order to approve the biocompatibility of nano bioactive glass. MATERIALS AND METHODS: In this in vitro experimental study, periodontal C165 fibroblasts cells were cultured in their logarithmic phase and the genotoxicity of novel synthesized bioactive glass nanoparticles and Novabone bioglass was studied in different concentrations and a control group using Comet assay test. By using Autocomet software, three parameters (Tail length, %DNA in tail, Tail moment) were analyzed; the genotoxicity of mentioned biomaterials and control group. Obtained data were analyzed by SPSS 11.5 software, Kruskal Wallis H and Mann Whitney tests (P = 0.05). RESULTS: No statistically significant difference was observed between the concentrations of Novabone bioglass (P value = 0.085) with control group and novel nano bioactive glass (P value = 0.437) with control group in the evaluation of %DNA in tail parameter. There was significant difference between genotoxicity of novel nano bioactive glass and control, and between Novabone bioglass and control group in concentrations of 4 and 5 mg/ml. According to significance of the mean difference, novel nano bioactive glass showed higher genotoxicity compared to Novabone bioglass in the concentration of 5 mg/ml (P ≤ 0.05). CONCLUSION: The findings of this study have demonstrated that novel nano bioactive glass had no genotoxicity in concentrations lower than 4 mg/ml. Nanoparticles have a higher surface area in comparison to microparticles and thus, the amount and rate of ion release for nanoparticles are extremely higher. This difference is the main reason for the different genotoxicity of nano bioactive glass and micro Novabone bioglass in the concentrations higher than 4 mg/ml.

New approach to bone tissue engineering: simultaneous application of hydroxyapatite and bioactive glass coated on a poly(L-lactic acid) scaffold.

A combination of bioceramics and polymeric nanofibers holds promising potential for bone tissue engineering applications. In the present study, hydroxyapatite (HA), bioactive glass (BG), and tricalcium phosphate (TCP) particles were coated on the surface of electrospun poly(L-lactic acid) (PLLA) nanofibers, and the capacity of the PLLA, BG-PLLA, HA-PLLA, HA-BG-PLLA, and TCP-PLLA scaffolds for bone regeneration was investigated in rat critical-size defects using digital mammography, multislice spiral-computed tomography (MSCT) imaging, and histological analysis. Electrospun scaffolds exhibited a nanofibrous structure with a homogeneous distribution of bioceramics along the surface of PLLA nanofibers. A total of 8 weeks after implantation, no sign of complication or inflammation was observed at the site of the calvarial bone defect. On the basis of imaging analysis, a higher level of bone reconstruction was observed in the animals receiving HA-, BG-, and TCP-coated scaffolds compared to an untreated control group. In addition, simultaneous coating of HA and BG induced the highest regeneration among all groups. Histological staining confirmed these findings and also showed an efficient osseointegration in HA-BG-coated nanofibers. On the whole, it was demonstrated that nanofibrous structures could serve as an appropriate support to guide the healing process, and coating their surface with bioceramics enhanced bone reconstruction. These bioceramic-coated scaffolds can be used as new bone-graft substitutes capable of efficiently inducing osteoconduction and osseointegration in orthopedic fractures and defects.

Cytotoxicity evaluation of 63s bioactive glass and bone-derived hydroxyapatite particles using human bone-marrow stem cells.

BACKGROUND: In recent years, bioceramics have been favored by biomaterials scientists and researchers. Due to their special and distinctive features, bioactive glass and hydroxyapatite possess a higher place among different types of bioceramics. METHOD: In this study, the effect of 63S bioactive glass and bone-derived hydroxyapatite particles on the proliferation of human bone-marrow stem cells (hMSCs) was investigated. Bioactive glass particles were made via sol-gel method and hydroxyapatite was obtained from bovine bone. The particle size and morphology were investigated by scanning electron microscope (SEM). Then the in vitro cytotoxicity of particles was evaluated using MTT assay. SEM showed that bioactive glass particles were in the nanoscale range and had tendency towards agglomeration. It was also confirmed that the hydroxyapatite particles were agglomerations of crystals cca 50-500 nm across. RESULTS: The results of MTT assay confirmed the viability and proliferation of hMSCs in contact with bioactive glass and bone-derived HA particles. The fabricated particles in combination with stem cells were shown to hold promising potential for further applications in tissue engineering and regenerative medicine.