Synthesis, Characterization and Osteogenic Properties of Chitin-Polydioxanone Composite Gel.

In this study, we have developed a Chitin(Ch)-Poly(dioxanone)(PDO) gel system, which can be potentially used for tissue engineering. Hydrogel has been widely used in biomedical applications for its tuneable properties and biocompatibility. Chitin (Ch) is a natural biopolymer used for its ability to mimic the natural extracellular matrix due to N-acetyl glucosamine structural units. Poly (dioxanone) (PDO) is an FDA-approved synthetic biopolymer known for its mechanical properties, good biocompatibility, and poor inflammatory response. Based on this, we have developed Ch-PDO composite gel using simple regeneration chemistry and characterized it using FT-IR and SEM. The developed composite gel showed improved gel strength, good swelling ability,and controlled degradation behaviour. It also showed good injectability with shear thinning properties and hemocompatibility. Further, the biocompatibility and cell adhesion studies of the prepared gels were studied using dental follicle stem cells (DFSCs). The prepared Ch-PDO gel was biocompatible and showed DFSCs cell attachment. Osteogenic mineralization and RUNX2 expression of the prepared Ch and Ch-PDO gel was studied and Ch-PDO gel showed an enhanced mineralization and RUNX2 expression. Therefore, the developed chitin-PDO gel could be potentially used for bone tissue engineering.

Mechanical properties of tissue adhesives used for retaining extraoral silicone prostheses in maxillofacial defects: A systematic review.

STATEMENT OF PROBLEM: Different tissue adhesives are available for retaining extraoral silicone prostheses in maxillofacial defects. Comparative assessment of their mechanical properties will help the clinician select the right product for a specific clinical situation, but a systematic review is lacking. PURPOSE: The purpose of this systematic review was to analyze the existing data in the literature regarding 5 mechanical properties of tissue adhesives for extraoral silicone prostheses: peel strength, tensile strength, shear strength, torsional strength, and tack. MATERIAL AND METHODS: A manual and electronic search was performed in appropriate databases to identify relevant publications with specific inclusion and exclusion criteria. The retrieved studies were screened for eligibility using the title, abstract, and published full texts. To evaluate the risk of bias, a methodological quality assessment was performed using the Joanna Briggs Institute Critical Appraisal Checklists for Randomized Controlled Trials. A custom data extraction template was used, and the results pooled using descriptive methods. RESULTS: After screening, 15 articles, 5 in vitro studies, and 10 clinical studies were eligible for data extraction. In vitro studies assessed tack and peel bond strength. In contrast, clinical studies assessed peel strength, tensile strength, shear strength, and torsional strength through direct comparisons and in diverse settings. CONCLUSIONS: The assessed studies showed considerable methodological heterogeneity. When silicone-based tissue adhesives (Secure2 Medical Adhesive; Factor II, Hollister Colostomy Adhesive; Hollister Inc, Dow Corning 355 Medical adhesives; Dow Corning Europe Inc) were compared with water-based adhesives (Pros-Aide Adhesive; ADM Tronics Inc, PSA 1; Cosmedica Ltd, Daro adhesive; Factor II, Epithane-3; Daro Products), the silicone-based adhesives showed a higher peel bond strength (PBS), while double-sided medical adhesive tapes showed lower PBS. A few studies evaluated variations in the PBS as being affected by the addition of stone wool fibers, immersion in water, application of skin protective dressings and adhesive removers, application of multiple layers of adhesive, and usage of urethane liner.

Amikacin sulphate loaded chitosan-diopside nanoparticles composite scaffold for infectious wound healing.

A wound dressing material should inhibit infections that may occur at the wound site, and at the same time, it should enhance the healing process. In this study, we developed an amikacin sulphate (AK) incorporated chitosan (Ch) and Diopside nanoparticles composite dressing (Ch-nDE-AK) for controlling wound infection and healing. The diopside nanoparticles (nDE) were prepared using sol-gel synthesis and characterized using XRD, FT-IR, and FESEM. nDE shows a size range of 142 ± 31 nm through FESEM analysis. Later, the developed composite dressing was characterized using SEM, EDS, and FT-IR analysis. Ch-nDE-AK dressing possesses a porous nature that will aid in easy cell infiltration and proliferation. The swelling studies indicated the expansion capability of the scaffold when applied to the injured site. Ch-nDE-AK scaffold showed a 69.6 ± 8.2 % amikacin sulphate release up to 7 days, which indicates the sustained release of the drug from Ch-nDE-AK scaffold. The drug release data was subjected to various kinetics models and was observed to follow the Higuchi model. The scaffold showed antibacterial activity against ATCC strains of S. aureus and E. coli for 7 days by in vitro. Ch-nDE-AK scaffold also showed antibacterial activity against S. aureus and E. coli clinical strains in vitro. The ex vivo antibacterial study confirmed the antibacterial ability of Ch-nDE-AK scaffold against S. aureus and E. coli. Ch-nDE-AK scaffold also exhibits anti-biofilm activity against S. aureus and E. coli. The Ch-nDE-AK scaffold showed cytocompatibility and cell attachment to fibroblast cells. Additionally, the scratch assay using fibroblast cells confirmed the role of the nDE in the scaffold, helping in cell migration. Thus, the developed Ch-nDE-AK dressing can potentially be used to treat infectious wound healing.

Lyophilized platelet rich fibrin and gelatin incorporated bioadhesive bone cement composite for repair of mandibular continuity defects.

Mandibular continuity defects stem from conditions such as malignancies, trauma, cysts, osteomyelitis and osteoradionecrosis, presenting significant challenges. If mandibular reconstruction fails, it can result in facial collapse, causing significant aesthetic and functional concerns for the patient. In the present study we developed a bio-adhesive Bone Cement (BC) enriched with lyophilised PRF and gelatin to enhance bone repair and induce regeneration. The developed BC consisted of a mixture of Tetracalcium Phosphate (TTCP) and O-Phospho-l-serine (OPLS) in addition to lyophilised Platelet Rich Fibrin (PRF) for sustained growth factor release and gelatin (GE) for improved cement resorption. It is primarily designed for in-situ application, conforming to the shape and size of the defect for effective bone repair and regeneration. The study evaluated four groups: (i) BC (control), (ii) BC-GE (control), (iii) BC-PRF, and (iv) BC-GE-PRF. All the four groups were characterised using FTIR, SEM and XRD. The mechanical studies of the prepared beads exhibited a significant increase in the compressive strength of the PRF loaded bone cement composites. In vitro degradation study of the beads over a 60-day period revealed a significantly higher percentage of bone cement resorption in the gelatin-incorporated groups, BC-GE (44 ± 0.5 %), and BC-GE-PRF (45 ± 2 %). The assessment of growth factor release (TGF-ß and VEGF) using ELISA revealed a prolonged and sustained release of both growth factors over a 28-day period. In vitro studies were performed on human Dental Follicle Stem Cells (DFSCs) to assess cell attachment, proliferation, mineralisation and osteogenic differentiation. These studies clearly depicted that BC-PRF and BC-GE-PRF showed significantly greater proliferation of DFSCs. Furthermore, BC-PRF and BC-GE-PRF samples exhibited notably elevated expression of Runx2 and OPN (osteogenic markers), as well as a higher intensity of alizarin red stain (mineralisation). Therefore, it was concluded that PRF incorporated bioadhesive bone cement composites greatly enhance the cell attachment, proliferation, mineralisation and osteogenic differentiation of the DFSCs. Thus, the PRF and gelatin incorporated bone cement composites is expected to facilitate effective and faster bone regeneration and healing in a wide range of dental and maxillofacial defects.

Diabetic Pedal Osteomyelitis and Its Treatment.

Diabetes is a fast-growing chronic metabolic disorder that is widely associated with foot ulcers. The major challenge among these ulcers is wound infections, altered inflammatory responses, and a lack of angiogenesis that can complicate limb amputation. The foot, because of its architecture, becomes the part most prone to complications and the infection rate is higher mainly between the toes due to the humid nature. Therefore, the infection rate is significantly higher. Wound healing in diabetes is a dynamic process usually delayed due to poor immune function. Diabetes-related pedal neuropathy and perfusion disturbances can lead to a loss of sensation in the foot. This neuropathy can further be a risk factor for ulcer development due to repetitive mechanical stress that later might get infected by the invasion of microorganisms extending to the bone and causing an infection called pedal osteomyelitis. This review details the pathophysiology, the biomaterials aiding in the infection cure and regeneration of bone along with their limitations, as well as their future prospects.

Trends in mechanobiology guided tissue engineering and tools to study cell-substrate interactions: a brief review.

Sensing the mechanical properties of the substrates or the matrix by the cells and the tissues, the subsequent downstream responses at the cellular, nuclear and epigenetic levels and the outcomes are beginning to get unraveled more recently. There have been various instances where researchers have established the underlying connection between the cellular mechanosignalling pathways and cellular physiology, cellular differentiation, and also tissue pathology. It has been now accepted that mechanosignalling, alone or in combination with classical pathways, could play a significant role in fate determination, development, and organization of cells and tissues. Furthermore, as mechanobiology is gaining traction, so do the various techniques to ponder and gain insights into the still unraveled pathways. This review would briefly discuss some of the interesting works wherein it has been shown that specific alteration of the mechanical properties of the substrates would lead to fate determination of stem cells into various differentiated cells such as osteoblasts, adipocytes, tenocytes, cardiomyocytes, and neurons, and how these properties are being utilized for the development of organoids. This review would also cover various techniques that have been developed and employed to explore the effects of mechanosignalling, including imaging of mechanosensing proteins, atomic force microscopy (AFM), quartz crystal microbalance with dissipation measurements (QCMD), traction force microscopy (TFM), microdevice arrays, Spatio-temporal image analysis, optical tweezer force measurements, mechanoscanning ion conductance microscopy (mSICM), acoustofluidic interferometric device (AID) and so forth. This review would provide insights to the researchers who work on exploiting various mechanical properties of substrates to control the cellular and tissue functions for tissue engineering and regenerative applications, and also will shed light on the advancements of various techniques that could be utilized to unravel the unknown in the field of cellular mechanobiology.

In-situ silver nanoparticles incorporated N, O-carboxymethyl chitosan based adhesive, self-healing, conductive, antibacterial and anti-biofilm hydrogel.

Hydrogels are excellent wound healing materials. However, due to the wear and tear at the wound site, hydrogels can lose their structural and functional integrity. To overcome this and to effectively seal the wound and control infection, an in-situ silver nanoparticles (AgNps) incorporated N, O-carboxymethyl chitosan (N, O-CMC) based self-healing hydrogel using ethylenediaminetetraacetic acid-ferric ion (EDTA: Fe3+) complex was developed. The prepared N, O-CMC/AgNps hydrogel was characterized using FTIR, SEM, and TEM. The developed N, O-CMC/AgNps hydrogel was found to be adhesive, injectable, conductive, bio-compatible, and showed antibacterial activity against ATCC and clinical strains of E. coli, K. pneumonia, P. aeruginosa, S. aureus and MRSA. N, O-CMC/AgNps hydrogel also showed anti-biofilm activity against S. aureus, E. coli, and P. aeruginosa (ATCC strains). This developed antibacterial and self-healing N, O-CMC/AgNps hydrogel can be used in the treatment of infected wounds.

Combinatorial effect of plasma treatment, fiber alignment and fiber scale of poly (ε-caprolactone)/collagen multiscale fibers in inducing tenogenesis in non-tenogenic media.

Tendon being a hypocellular, low vascularized tissue often requires assistance for restoration after complete tear. Tendon tissue engineering aims in the development of suitable scaffold that could support the regeneration of tendon after damage. The success of such scaffolds is dependent on its integration with the native tissue which in turn is influenced by the cell-material interaction. In this work aligned poly(ε-caprolactone)/collagen (PCL/collagen) multiscale fibers were developed and plasma treatment using argon, nitrogen and its combination was accessed for inducing tenogenic differentiation in mesenchymal stem cells. The developed fibers mimicked tendon extracellular matrix (ECM) which upon plasma treatment maintained moderate hydrophilicity. Oxygen and nitrogen containing groups were observed to be incorporated after argon and nitrogen treatment respectively. Statistically significant (p < 0.001) enhancement was observed in average and root mean square (RMS) roughness after plasma treatment with the maximum in argon treated fibers. Vitronectin was competitively (statistically significant, p < 0.05) adsorbed after argon and combination treatment whereas nitrogen treatment led to the competitive adsorption of fibronectin (statistically significant, p < 0.05). Human mesenchymal stem cells (hMSCs) showed enhanced proliferation and attachment on plasma treated fibers. Increased porosity due to the presence of sacrificial collagen nanofibers improved cell infiltration which was further enhanced upon plasma treatment. RhoA activation was observed (statistically significant, p < 0.05) on aligned PCL/collagen multiscale fibers and PCL microfibers, which proved its impact on tenogenic differentiation. Further enhancement in rhoA expression was observed on argon (p < 0.01) and combination plasma (p < 0.05) treated fibers. Tenogenic differentiation of hMSCs was enhanced (statistically significant) on argon plasma treated aligned fibers which was confirmed by the expression of scleraxis, mohawk (early markers) and tenomodulin (late marker) at protein level and mohawk, collagen I, collagen III (early markers), thrombospondin 4 and tenascin C (late markers) at gene level. Thus argon plasma treatment on aligned fibers is an effective method to induce tenogenesis even in non-tenogenic media.

Addition of lactoferrin and substance P in a chitin/PLGA-CaSO4 hydrogel for regeneration of calvarial bone defects.

Calcium-based injectable hydrogels with various bioactive active molecules possess a great potential for bone regeneration. Herein, we have synthesized a chitin-PLGA-calcium sulfate hydrogel (CSG) containing bioactive molecules - lactoferrin (LF) and substance P (SP). SEM and XRD analysis revealed that CS crystal growth was altered with the addition of LF. Rheological measurements indicated that the injectability of the hydrogels was maintained after the addition of LF, however, there was a reduction in storage modulus after LF addition. The addition of LF increased stem cell proliferation whereas, SP enhanced the cell migration. Osteogenic gene expression revealed that LF concentration at 25 µg/mg of CSG was optimal for a favourable outcome. To this optimized LF containing CSG, SP was incorporated and 0.05 µg/mg was found to be most effective (CSG-L3S2) in vitro studies. Further, the µ-CT and histological studies confirmed that CSG-L3S2 showed enhanced bone regeneration compared to the controls in critical-sized calvarial defect of mice. Thus the results indicate that a combination of the chemotactic agent (SP), pleiotropic growth protein (LF), and CS in the chitin-PLGA hydrogel could be a promising approach for non-load bearing bone defects.

Vasoconstrictor and coagulation activator entrapped chitosan based composite hydrogel for rapid bleeding control.

Chitosan (Cs) as a hemostatic agent has been in use to control hemorrage. Composite hydrogel formed by entrapment of vasoconstrictor-potassium aluminium sulfate (0.25 %PA) and coagulation activator-calcium chloride (0.25 %Ca) into Cs (2 %) hydrogel would enhance the hemostatic property of Cs. In this work, the prepared composite hydrogel was injectable, shear thinning, cyto and hemocompatible. The 2 %Cs-0.25 %PA-0.25 %Ca composite hydrogel caused rapid blood clotting by accelerating RBC/platelet aggregation and activation of the coagulation cascade. Further, in vivo studies on rat liver and femoral artery hemorrage model showed the efficiency of 2 %Cs-0.25 %PA-0.25 %Ca composite hydrogel to achieve hemostasis in a shorter time (20 ± 10 s, 105 ± 31 s) than commercial hemostatic agents-Fibrin sealant (77 ± 26 s, 204 ± 58 s) and Floseal (76 ± 15 s, 218 ± 46 s). In in vivo toxicological study, composite hydrogel showed material retention even after 8 weeks post-surgery, therefore excess hydrogel should be irrigated from site of application. This prepared composite hydrogel based hemostatic agent has potential application in low pressure bleeding sites.

Nanocurcumin and arginine entrapped injectable chitosan hydrogel for restoration of hypoxia induced endothelial dysfunction.

Hypoxia is a condition that gradually leads to ischemic damages in organs which is marked by poor tissue perfusion. Depending on the severity of the condition, revascularisation therapies are needed for reducing the risk of organ dysfunction. This study was aimed at developing an injectable nanocurcumin and arginine incorporated chitosan hydrogel (nC/R) that can prevent hypoxia induced endothelial damage. The prepared hydrogel has shear thinning, stable and injectable nature. The (nC and nC/R) hydrogels showed significant antioxidant activity and biodegradation in vitro. The release of curucmin and arginine from the nC/R was found to be higher at acidic pH, which predominates in an ischemic site. To mimic low oxygen environment, an in vitro hypoxic endothelial dysfunction model was developed which showed decreased expressions of phosphorylated eNOS (serine 1177) when compared to the cells cultured in normoxic condition. In vitro tube formation assay demonstrated the protective effect of nC/R towards hypoxia induced reduction of tube width. The nC/R hydrogel was found to enhance phosphorylation of eNOS at serine 1177 site in cultured endothelial cells subjected to hypoxia. Therefore, nC/R hydrogel could effectively deliver both curcumin and arginine and therapeutically reduce the effect of hypoxia induced endothelial dysfunction.

Recent developments in controlling sternal wound infection after cardiac surgery and measures to enhance sternal healing.

One of the major risks of cardiac surgery is the occurrence of infection at the sternal wound site. Sternal wound infections are primarily classified into superficial infection and deep sternal wound infection or mediastinitis. A patient is diagnosed with mediastinitis if microorganisms are present in their mediastinal tissue/fluid or with the observation of sternal wound infection during operation and with characteristic symptoms including chest pain, fever, and purulent drainage from the mediastinum. It is usually caused by Staphylococcal organisms in 75.8% of cases and the rest is caused by gram-negative bacteria. Currently, in cardiac surgery, hemostasis is achieved using electrocautery and bone wax, and the sternum is closed using wire cerclage. Several studies show that bone wax can act as a nidus for initiation of infection and the oozing blood and hematoma at the site can promote the growth of infectious organisms. Many research groups have developed different types of biomaterials and reported on the prevention of infection and healing of the sternum. These materials are reported to have both positive and negative effects. In this review, we highlight the current clinical practices undertaken to prevent infection and bleeding as well as research progress in this field and their outcomes in controlling bleeding, infection, and enhancing sternal healing.

Human Adipose Tissue Derivatives as a Potent Native Biomaterial for Tissue Regenerative Therapies.

BACKGROUND: Human adipose tissue is a great source of translatable biomaterials owing to its ease of availability and simple processing. Reusing discardable adipose tissue for tissue regeneration helps in mimicking the exact native microenvironment of tissue. Over the past 10 years, extraction, processing, tuning and fabrication of adipose tissue have grabbed the attention owing to their native therapeutic and regenerative potential. The present work gives the overview of next generation biomaterials derived from human adipose tissue and their development with clinical relevance. METHODS: Around 300 articles have been reviewed to widen the knowledge on the isolation, characterization techniques and medical applications of human adipose tissue and its derivatives from bench to bedside. The prospective applications of adipose tissue derivatives like autologous fat graft, stromal vascular fraction, stem cells, preadipocyte, adipokines and extracellular matrix, their behavioural mechanism, rational property of providing native bioenvironment, circumventing their translational abilities, recent advances in featuring them clinically have been reviewed extensively to reveal the dormant side of human adipose tissue. RESULTS: Basic understanding about the molecular and structural aspect of human adipose tissue is necessary to employ it constructively. This review has nailed the productive usage of human adipose tissue, in a stepwise manner from exploring the methods of extracting derivatives, concerns during processing and its formulations to turning them into functional biomaterials. Their performance as functional biomaterials for skin regeneration, wound healing, soft tissue defects, stem cell and other regenerative therapies under in vitro and in vivo conditions emphasizes the translational efficiency of adipose tissue derivatives. CONCLUSION: In the recent years, research interest has inclination towards constructive tissue engineering and regenerative therapies. Unravelling the maximum utilization of human adipose tissue derivatives paves a way for improving existing tissue regeneration and cellular based therapies and other biomedical applications.

Biomimetic Materials and Fabrication Approaches for Bone Tissue Engineering.

Various strategies have been explored to overcome critically sized bone defects via bone tissue engineering approaches that incorporate biomimetic scaffolds. Biomimetic scaffolds may provide a novel platform for phenotypically stable tissue formation and stem cell differentiation. In recent years, osteoinductive and inorganic biomimetic scaffold materials have been optimized to offer an osteo-friendly microenvironment for the osteogenic commitment of stem cells. Furthermore, scaffold structures with a microarchitecture design similar to native bone tissue are necessary for successful bone tissue regeneration. For this reason, various methods for fabricating 3D porous structures have been developed. Innovative techniques, such as 3D printing methods, are currently being utilized for optimal host stem cell infiltration, vascularization, nutrient transfer, and stem cell differentiation. In this progress report, biomimetic materials and fabrication approaches that are currently being utilized for biomimetic scaffold design are reviewed.

In vitro targeted imaging and delivery of camptothecin using cetuximab-conjugated multifunctional PLGA-ZnS nanoparticles.

BACKGROUND: Targeted cancer therapy has been extensively developed to improve the quality of treatment by reducing the systemic exposure of cytotoxic drug. Polymeric nanoparticles with conjugated targeting agents are widely investigated because they offer tunability in particle size, drug release profile and biocompatibility. MATERIALS & METHODS: Here, we have prepared targeted multifunctional nanoparticles composed of a poly(lactic-co-glycolic acid) matrix, ZnS:Mn(2+) quantum dots and camptothecin, and targeted them to EGF receptor overexpressing cells with a cetuximab antibody. RESULTS: Physicochemical characterization of multifunctional nanoparticles showed stable particles with sizes of <200 nm. In vitro drug release and blood contact studies showed a sustained release profile, with limited hemolysis. In vitro cytotoxicity and cell uptake studies were carried out in A549, KB and MFC-7 cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, FACS, fluorescent microscopic images and spectroflourimetry. CONCLUSION: Our studies revealed higher camptothecin activity and uptake in cell lines that overexpress the EGF receptor. All these results suggest that anti-EGF receptor cetuximab-conjugated poly(lactic-co-glycolic acid) multifunctional nanoparticles can be used as a potential nanomedicine against cancer.

Multifunctional chitin nanogels for simultaneous drug delivery, bioimaging, and biosensing.

In this work, we developed biodegradable chitin nanogels (CNGs) by controlled regeneration method. For multifunctionalization, we have conjugated CNGs with MPA-capped-CdTe-QDs (QD-CNGs) for the in vitro cellular localization studies. In addition, the Bovine Serum Albumin (BSA) was loaded on to QD-CNGs (BSA-QD-CNGs). The CNGs, QD-CNGs, and BSA-QD-CNGs were well-characterized by SEM and AFM, which shows that the nanogels are in the range of <100 nm. These were further characterized by FT-IR and Cyclic Voltametry. The cytocompatibility assay showed that the nanogels are nontoxic to L929, NIH-3T3, KB, MCF-7, PC3, and VERO cells. The cell uptake studies of the QD-CNGs were analyzed, which showed retention of these nanogels inside the cells (L929, PC3, and VERO). In addition, the protein loading efficiency of the nano gels has also been analyzed. Our preliminary studies reveal that these multifunctionalized nanogels could be useful for drug delivery with simultaneous imaging and biosensing.