Current Perspectives of Protein in Bone Tissue Engineering: Bone Structure, Ideal Scaffolds, Fabrication Techniques, Applications, Scopes, and Future Advances.

In view of their exceptional approach, excellent inherent biocompatibility and biodegradability properties, and interaction with the local extracellular matrix, protein-based polymers have received attention in bone tissue engineering, which is a multidisciplinary field that repairs and regenerates fractured bones. Bone is a multihierarchical complex structure, and it performs several essential biofunctions, including maintaining mineral balance and structural support and protecting soft organs. Protein-based polymers have gained interest in developing ideal scaffolds as emerging biomaterials for bone fractured healing and regeneration, and it is challenging to design ideal bone substitutes as perfect biomaterials. Several protein-based polymers, including collagen, keratin, gelatin, serum albumin, etc., are potential materials due to their inherent cytocompatibility, controlled biodegradability, high biofunctionalization, and tunable mechanical characteristics. While numerous studies have indicated the encouraging possibilities of proteins in BTE, there are still major challenges concerning their biodegradability, stability in physiological conditions, and continuous release of growth factors and bioactive molecules. Robust scaffolds derived from proteins can be used to replace broken or diseased bone with a biocompatible substitute; proteins, being biopolymers, provide excellent scaffolds for bone tissue engineering. Herein, recent developments in protein polymers for cutting-edge bone tissue engineering are addressed in this review within 3-5 years, with a focus on the significant challenges and future perspectives. The first section discusses the structural fundamentals of bone anatomy and ideal scaffolds, and the second section describes the fabrication techniques of scaffolds. The third section highlights the importance of proteins and their applications in BTE. Hence, the recent development of protein polymers for state-of-the-art bone tissue engineering has been discussed, highlighting the significant challenges and future perspectives.

Fabrication of Bilayer Nanofibrous-Hydrogel Scaffold from Bacterial Cellulose, PVA, and Gelatin as Advanced Dressing for Wound Healing and Soft Tissue Engineering.

Tissue engineering is currently one of the fastest-growing areas of engineering, requiring the fabrication of advanced and multifunctional materials that can be used as scaffolds or dressings for tissue regeneration. In this work, we report a bilayer material prepared by electrospinning a hybrid material of poly(vinyl alcohol) (PVA) and bacterial cellulose (BC NFs) (top layer) over a highly interconnected porous 3D gelatin-PVA hydrogel obtained by a freeze-drying process (bottom layer). The techniques were combined to produce an advanced material with synergistic effects on the physical and biological properties of the two materials. The bilayer material was characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and a water contact measurement system (WCMS). Studies on swelling, degradability, porosity, drug release, cellular and antibacterial activities were performed using standardized procedures and assays. FTIR confirmed cross-linking of both the top and bottom layers, and SEM showed porous structure for the bottom layer, random deposition of NFs on the surface, and aligned NFs in the cross section. The water contact angle (WCA) showed a hydrophilic surface for the bilayer material. Swelling analysis showed high swelling, and degradation analysis showed good stability. The bilayer material released Ag-sulfadiazine in a sustained and controlled manner and showed good antibacterial activities against severe disease-causing gram + ive and -ive (Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa) bacterial strains. In vitro biological studies were performed on fibroblasts (3T3) and human embryonic kidneys (HEK-293), which showed desirable cell viability, proliferation, and adhesion to the bilayer. Thus, the synergistic effect of NFs and the hydrogel resulted in a potential wound dressing material for wound healing and soft tissue engineering.

Effect Of Eight Hours Per Day Of Intermittent Self Prone Positioning For Seven Days On The Severity Of Covid-19 Pneumonia/ Acute Respiratory Distress Syndrome.

BACKGROUND: Prone positioning improves ventilation-perfusion mismatch, distribution of gravitational gradient in pleural pressure, and oxygen saturation significantly in patients with Covid pneumonia. We aimed to find out the efficacy of eight hours per day of intermittent selfprone positioning for seven days in patients affected with COVID-19 pneumonia/ ARDS. METHODS: This Randomized Clinical Trial was conducted in the Covid isolation wards of Ayub Teaching Hospital, Abbottabad. Patients suffering from COVID-19 pneumonia/ ARDS were enrolled with permuted block randomization into a control and an experimental group each consisting of 36 patients. Parameters of Pneumonia Severity Index (PSI) score along with other sociodemographic data was noted on a preformed structured questionnaire. Death was confirmed by requesting the death certificate of patients on the 90th day of enrolment. Data Analysis was done with SPSS Version 25. Tests of significance were applied to calculate the difference in the patients of the two groups with respect to respiratory physiology and survival. RESULTS: The mean age of the patients was 63.79±15.26 years. A total of 25 (32.9%) male and 47 (61.8%) female patients were enrolled. Statistically significant improvement was found in the respiratory physiology of the patients at 7th and 14th DOA between the groups. Pearson Chi-Square test of significance showed a difference in mortality between the two groups at 14th DOA (pvalue=0.011) but not at 90th DOA (p-value=0.478). Log Rank (Mantel-Cox) test of significance, applied on the Kaplan Meier curve and showed no statistically significant difference among the groups based on the survival of the patients. (p-value=0.349). CONCLUSIONS: Early transient improvement in respiratory physiology and mortality does occur with 8 hours of self-prone positioning for seven days but there is no effect on the 90-day survival of the patients. Thus, the impact of the manoeuvre on improving survival needs to be explored with studies having an application of the manoeuvre for a longer duration and period.

Artificial neural networks for the prediction of biochar yield: A comparative study of metaheuristic algorithms.

In this study, an integrated framework of artificial neural networks (ANNs) and metaheuristic algorithms have been developed for the prediction of biochar yield using biomass characteristics and pyrolysis process conditions. Comparative analysis of six different metaheuristic algorithms was performed to optimize the ANN architecture and select important features. The results suggested that the ANN model coupled with the Rao-2 algorithm outperformed (R2 âˆ¼ 0.93, RMSE âˆ¼ 1.74%) all other models. Furthermore, the detailed information behind the models was acquired, identifying the most influencing factors as follows: pyrolysis temperature (56%), residence time (23%), and heating rate (8%). The partial dependence plot analysis revealed how each influencing factor affected the target variable. Finally, an easy-to-use software tool for predicting biochar yield was built using the ANN-Rao-2 model. This study demonstrates huge potential that machine learning presents in predictive modelling of complex pyrolysis processes, and reduces the time-consuming and expensive experimental work for estimating the biochar yield.

Fabrication and characterization of quercetin loaded casein phosphopeptides-chitosan composite nanoparticles by ultrasound treatment: Factor optimization, formation mechanism, physicochemical stability and antioxidant activity.

Ultrasound treatment was used to successfully prepare Quercetin (Qu)-loaded Casein phosphopeptides (CPP)/chitosan (CS) nanoparticles. Compared with the control, the above ternary nanoparticles with the smallest size (241.27 nm, decreased by 34.32%), improved encapsulation efficiency of Qu (78.55%, increased by 22.12%) when prepared under following conditions: ultrasonic frequency, 20/35/50 kHz; the power density, 80 W/L; the time, 20 min, and the intermittent ratio, 20 s/5s. Electrostatic interactions, hydrogen bonding, and hydrophobic interactions were the main driving forces for nanoparticles formulation, which were strengthened by ultrasound treatment. The compact, homogeneous and spherical composite nanoparticles obtained by sonication were clearly observed by scanning electron microscope and atomic force microscope. The environmental stability (NaCl, pH, exposure time, storage time, and simulated gastrointestinal digestion) and antioxidant activity of the ternary nanoparticles were remarkably enhanced after ultrasonic treatment. Furthermore, the ternary nanoparticles prepared by ultrasound exhibited excellent stability in simulated gastrointestinal digestion. The above results indicate that ultrasound not only increases the loading of the nanoparticles on bioactive substances but also improves the environmental stability and antioxidant activity of the formed nanoparticles. Ultrasound-assisted preparation of nanoparticles loaded with bioactive substances could be well used in the functional food and beverage industry.

Development of Biopolymeric Hybrid Scaffold-Based on AAc/GO/nHAp/TiO2 Nanocomposite for Bone Tissue Engineering: In-Vitro Analysis.

Bone tissue engineering is an advanced field for treatment of fractured bones to restore/regulate biological functions. Biopolymeric/bioceramic-based hybrid nanocomposite scaffolds are potential biomaterials for bone tissue because of biodegradable and biocompatible characteristics. We report synthesis of nanocomposite based on acrylic acid (AAc)/guar gum (GG), nano-hydroxyapatite (HAp NPs), titanium nanoparticles (TiO2 NPs), and optimum graphene oxide (GO) amount via free radical polymerization method. Porous scaffolds were fabricated through freeze-drying technique and coated with silver sulphadiazine. Different techniques were used to investigate functional group, crystal structural properties, morphology/elemental properties, porosity, and mechanical properties of fabricated scaffolds. Results show that increasing amount of TiO2 in combination with optimized GO has improved physicochemical and microstructural properties, mechanical properties (compressive strength (2.96 to 13.31 MPa) and Young's modulus (39.56 to 300.81 MPa)), and porous properties (pore size (256.11 to 107.42 µm) and porosity (79.97 to 44.32%)). After 150 min, silver sulfadiazine release was found to be ~94.1%. In vitro assay of scaffolds also exhibited promising results against mouse pre-osteoblast (MC3T3-E1) cell lines. Hence, these fabricated scaffolds would be potential biomaterials for bone tissue engineering in biomedical engineering.

A Review on Current Trends of Polymers in Orthodontics: BPA-Free and Smart Materials.

Polymeric materials have always established an edge over other classes of materials due to their potential applications in various fields of biomedical engineering. Orthodontics is an emerging field in which polymers have attracted the enormous attention of researchers. In particular, thermoplastic materials have a great future utility in orthodontics, both as aligners and as retainer appliances. In recent years, the use of polycarbonate brackets and base monomers bisphenol A glycerolate dimethacrylate (bis-GMA) has been associated with the potential release of bisphenol A (BPA) in the oral environment. BPA is a toxic compound that acts as an endocrine disruptor that can affect human health. Therefore, there is a continuous search for non-BPA materials with satisfactory mechanical properties and an esthetic appearance as an alternative to polycarbonate brackets and conventional bis-GMA compounds. This study aims to review the recent developments of BPA-free monomers in the application of resin dental composites and adhesives. The most promising polymeric smart materials are also discussed for their relevance to future orthodontic applications.

Arabinoxylan/graphene-oxide/nHAp-NPs/PVA bionano composite scaffolds for fractured bone healing.

The importance of bone scaffolds has increased many folds in the last few years; however, during bone implantation, bacterial infections compromise the implantation and tissue regeneration. This work is focused on this issue while not compromising on the properties of a scaffold for bone regeneration. Biocomposite scaffolds (BS) were fabricated via the freeze-drying technique. The samples were characterized for structural changes, surface morphology, porosity, and mechanical properties through spectroscopic (Fourier transform-infrared [FT-IR]), microscopic (scanning electron microscope [SEM]), X-ray (powder X-ray diffraction and energy-dispersive X-ray), and other analytical (Brunauer-Emmett-Teller, universal testing machine Instron) techniques. Antibacterial, cellular, and hemocompatibility assays were performed using standard protocols. FT-IR confirmed the interactions of all the components. SEM illustrated porous and interconnected porous morphology. The percentage porosity was in the range of 49.75%-67.28%, and the pore size was 215.65-470.87 µm. The pore size was perfect for cellular penetration. Thus, cells showed significant proliferation onto these scaffolds. X-ray studies confirmed the presence of nanohydroxyapatite and graphene oxide (GO). The cell viability was 85%-98% (BS1-BS3), which shows no significant toxicity of the biocomposite. Furthermore, the biocomposites exhibited better antibacterial activity, no effect on the blood clotting (normal in vitro blood clotting), and less than 5% hemolysis. The ultimate compression strength for the biocomposites increased from 4.05 to 7.94 with an increase in the GO content. These exciting results revealed that this material has the potential for possible application in bone tissue engineering.

Recent Advances in Biopolymeric Composite Materials for Tissue Engineering and Regenerative Medicines: A Review.

The polymeric composite material with desirable features can be gained by selecting suitable biopolymers with selected additives to get polymer-filler interaction. Several parameters can be modified according to the design requirements, such as chemical structure, degradation kinetics, and biopolymer composites' mechanical properties. The interfacial interactions between the biopolymer and the nanofiller have substantial control over biopolymer composites' mechanical characteristics. This review focuses on different applications of biopolymeric composites in controlled drug release, tissue engineering, and wound healing with considerable properties. The biopolymeric composite materials are required with advanced and multifunctional properties in the biomedical field and regenerative medicines with a complete analysis of routine biomaterials with enhanced biomedical engineering characteristics. Several studies in the literature on tissue engineering, drug delivery, and wound dressing have been mentioned. These results need to be reviewed for possible development and analysis, which makes an essential study.

Correction: Development and in vitro evaluation of κ-carrageenan based polymeric hybrid nanocomposite scaffolds for bone tissue engineering.

[This corrects the article DOI: 10.1039/D0RA07446B.].

Development of Polymeric Nanocomposite (Xyloglucan-co-Methacrylic Acid/Hydroxyapatite/SiO2) Scaffold for Bone Tissue Engineering Applications-In-Vitro Antibacterial, Cytotoxicity and Cell Culture Evaluation.

Advancement and innovation in bone regeneration, specifically polymeric composite scaffolds, are of high significance for the treatment of bone defects. Xyloglucan (XG) is a polysaccharide biopolymer having a wide variety of regenerative tissue therapeutic applications due to its biocompatibility, in-vitro degradation and cytocompatibility. Current research is focused on the fabrication of polymeric bioactive scaffolds by freeze drying method for nanocomposite materials. The nanocomposite materials have been synthesized from free radical polymerization using n-SiO2 and n-HAp XG and Methacrylic acid (MAAc). Functional group analysis, crystallinity and surface morphology were investigated by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD) and scanning electron microscopy (SEM) techniques, respectively. These bioactive polymeric scaffolds presented interconnected and well-organized porous morphology, controlled precisely by substantial ratios of n-SiO2. The swelling analysis was also performed in different media at varying temperatures (27, 37 and 47 °C) and the mechanical behavior of the dried scaffolds is also investigated. Antibacterial activities of these scaffolds were conducted against pathogenic gram-positive and gram-negative bacteria. Besides, the biological behavior of these scaffolds was evaluated by the Neutral Red dye assay against the MC3T3-E1 cell line. The scaffolds showed interesting properties for bone tissue engineering, including porosity with substantial mechanical strength, biodegradability, biocompatibility and cytocompatibility behavior. The reported polymeric bioactive scaffolds can be aspirant biomaterials for bone tissue engineering to regenerate defecated bone.

Development and in vitro evaluation of κ-carrageenan based polymeric hybrid nanocomposite scaffolds for bone tissue engineering.

The excellent biocompatible and osteogenesis characteristics of porous scaffolds play a vital role in bone regeneration. In this study, we have synthesized polymeric hybrid nanocomposites via free-radical polymerization from carrageenan/acrylic-acid/graphene/hydroxyapatite. Porous hybrid nanocomposite scaffolds were fabricated through a freeze-drying method to mimic the structural and chemical composition of natural bone. Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and water contact-angle studies were carried-out for functional groups, surface morphology and hydrophilicity of the materials, followed by biodegradation and swelling analysis. The cell viability, cell culture and proliferation were evaluated against mouse pre-osteoblast (MC3T3-E1) cell lines using neutral red dye assay. The cell adherence and proliferation studies were determined by SEM. Physical characterization including optimum porosity and pore size (49.75% and 0.41 × 103 µm2), mechanical properties (compression strength 8.87 MPa and elastic modulus 442.63 MPa), swelling (70.20% at 27 °C and 77.21% at 37 °C) and biodegradation (23.8%) were performed. The results indicated CG-g-AAc-3 with a high optical density and better cell viability. Hence, CG-g-AAc-3 was found to be more efficient for bone regeneration with potential applications in fractured bone regeneration.

Problems in Mitochondrial DNA forensics: while interpreting length heteroplasmy conundrum of various Sindhi and Baluchi ethnic groups of Pakistan.

The insight heterodox genetics of mtDNA infer new perspectives at the level of human mitochondrial control region heteroplasmy, which is substantial in evolutionary as well as forensic interpretation. The main goal of this study is to interrogate the recurrence and resolve the ambiguity of blurry spectrum of heteroplasmy in the human mtDNA control region of 50 Baluchi and 116 Sindhi unrelated individuals. Sanger sequencing was employed classically, that was further investigated by minisequencing. Only 20% Baluchi and 25.8% Sindhi were homoplasmic, whereas rest of 80% Baluchi and 74.1% Sindhi exhibited at least one heteroplasmy within the specimen. In total, 166 individuals have length heteroplasmy (LH) found at positions 16189, 303-315, 568-573, and 514-524, whilst point mutation heteroplasmy (PMH) was detected at positions 73, 16093, 16189, and 16234, respectively. Overall LH was observed albeit high frequency in Sindhi ethnic group (82%) rather than Baluchi's (37%), whereas PMH accumulation was relatively extensive (24%) in Baluchi's than Sindhi's (11.2%). The obtained results ascertained that growing knowledge of heteroplasmy assisted to develop consciences in the forensic community that heteroplasmy plays a pivotal role in the legal interpretation on a regular basis and knowledge of its biological underpinnings has a vital niche in the forensic science. Limited studies have focused on heteroplasmy, yet scientific attention should be given, in order to determine its magnitude in different ethnic boundaries.