Spatiotemporally Controlled Release of Etamsylate from Bioinspired Peptide-Functionalized Nanoparticles Arrests Bleeding Rapidly and Improves Clot Stability in a Rabbit Internal Hemorrhage Model.

Achieving rapid clotting and clot stability are important unmet goals of clinical management of noncompressible hemorrhage. This study reports the development of a spatiotemporally controlled release system of an antihemorrhagic drug, etamsylate, in the management of internal hemorrhage. Gly-Arg-Gly-Asp-Ser (GRGDS) peptide-functionalized chitosan nanoparticles, with high affinity to bind with the GPIIa/IIIb receptor of activated platelets, were loaded with the drug etamsylate (etamsylate-loaded GRGDS peptide-functionalized chitosan nanoparticles; EGCSNP). Peptide conjugation was confirmed by LCMS, and the delivery system was characterized by DLS, SEM, XRD, and FTIR. In vitro study exhibited 90% drug release till 48 h fitting into the Weibull model. Plasma recalcification time and prothrombin time tests of GRGDS-functionalized nanoparticles proved that clot formation was 1.5 times faster than nonfunctionalized chitosan nanoparticles. The whole blood clotting time was increased by 2.5 times over clot formed under nonfunctionalized chitosan nanoparticles. Furthermore, the application of rheometric analysis revealed a 1.2 times stiffer clot over chitosan nanoparticles. In an in vivo liver laceration rabbit model, EGCSNP spatially localized at the internal injury site within 5 min of intravenous administration, and no rebleeding was recorded up to 3 h. The animals survived for 3 weeks after the injury, indicating the strong potential of the system for the management of noncompressible hemorrhage.

Stimulated Full-Thickness Cutaneous Wound Healing with Bioactive Dressings of Zinc and Cobalt Ion-Doped Bioactive Glass-Coated Eggshell Membranes in a Diabetic Rabbit Model.

Eggshell membrane-based biomedical applications have recently received great attention for their wound-healing properties. However, there are limited studies on diabetic wound healing. In this regard, we devised four types of composite eggshell membrane mats with nanoscale coatings of bioactive glass/Zn/Co-doped bioactive glass (ESM + BAG, ESM + ZnBAG, ESM + CoBAG, and ESM + ZnCoBAG) as wound-dressing materials for chronic nonhealing diabetic wounds. A detailed study of the physicochemical properties of the mats was conducted. In vitro studies demonstrated cytocompatibility and viability of human dermal fibroblasts on all four types of mats. The cells also attached finely on the mats with the help of cellular extensions, as evident from scanning electron microscopy (SEM) and rhodamine-phalloidin and Hoechst 33342 staining of cellular components. Endowed with bioactive properties, these mats influenced all aspects of full-thickness skin wound healing in diabetic animal model studies. All of the mats, especially the ESM + ZnCoBAG mat, showed the earliest wound closure, effective renewal, and restructuring of the extracellular matrix in terms of an accurate and timely accumulation of collagen, elastin, and reticulin fibers. Hydroxyproline and sulfated glycosaminoglycans were significantly (p < 0.01, p < 0.05) higher in ESM-ZnCoBAG-treated wounds in comparison to ESM-BAG-treated wounds, which suggests that these newly developed mats have potential as an affordable diabetic wound care solution in biomedical research.

Unveiling advanced strategies for therapeutic stem cell interventions in severe burn injuries: a comprehensive review.

This comprehensive review explores the complex terrain of stem cell therapies as a potential therapeutic frontier in the healing of complicated burn wounds. Serious tissue damage, impaired healing processes, and possible long-term consequences make burn wounds a complex problem. An in-depth review is required since, despite medical progress, existing methods for treating severe burn wounds have significant limitations. Burn wounds are difficult to heal because they cause extensive tissue damage. The challenges of burn injury-induced tissue regeneration and functional recovery are also the subject of this review. Although there is a lot of promise in current stem cell treatments, there are also some limitations with scalability, finding the best way to transport the cells, and finding consistent results across different types of patients. To shed light on how to improve stem cell interventions to heal severe burn wounds, this review covers various stem cell applications in burn wounds and examines these obstacles. To overcome these obstacles, one solution is to enhance methods of stem cell distribution, modify therapies according to the severity of the burn, and conduct more studies on how stem cell therapy affects individual patients. Novel solutions may also be possible through the combination of cutting-edge technologies like nanotechnology and biotechnology. This review seeks to increase stem cell interventions by analyzing present challenges and suggesting strategic improvements. The goal is to provide a more effective and tailored way to repair serious burn wounds.

Recent trends of stem cell therapies in The management of orthopedic surgical challenges.

Emerged health-related problems especially with increasing population and with the wider occurrence of these issues have always put the utmost concern and led medicine to outgrow its usual mode of treatment, to achieve better outcomes. Orthopedic interventions are one of the most concerning hitches, requiring advancement in several issues, that show complications with conventional approaches. Advanced studies have been undertaken to address the issue, among which stem cell therapy emerged as a better area of growth. The capacity of the stem cells to renovate themselves and adapt into different cell types made it possible to implement its use as a regenerative slant. Harvesting the stem cells, particularly mesenchymal stem cells is easier and can be further grown in vitro. In this review, we have discussed orthopedic-related issues including bone defects and fractures, non-unions, ligament and tendon injuries, degenerative changes, and associated conditions, which require further approaches to execute better outcomes, and the advanced strategies that can be tagged along with various ways of application of mesenchymal stem cells. It aims to objectify the idea of stem cells, with a major focus on the application of Mesenchymal stem cells (MSCs) from different sources in various orthopedic interventions. It also discusses the limitations, and future scopes for further approaches in the field of regenerative medicine. The involvement of mesenchymal stem cells may transition the procedures in orthopedic interventions from predominantly surgical substitution and reconstruction to bio-regeneration and prevention. Nevertheless, additional improvements and evaluations are required to explore the effectiveness and safety of mesenchymal stem cell treatment in orthopedic regenerative medicine.

Effects of the multiscale porosity of decellularized platelet-rich fibrin-loaded zinc-doped magnesium phosphate scaffolds in bone regeneration.

In recent years, metallic ion-doped magnesium phosphate (MgP)-based degradable bioceramics have emerged as alternative bone substitute materials owing to their excellent biocompatibility, bone-forming ability, bioactivity, and controlled degradability. Conversely, incorporating a biomolecule such as decellularized platelet-rich fibrin (d-PRF) on scaffolds has certain advantages for bone tissue regeneration, particularly in enhanced osteogenesis and angiogenesis. The present study focuses on the impact of d-PRF-loaded multiscale porous zinc-doped magnesium phosphate (Zn-MgP) scaffolds on biodegradability, biocompatibility, and bone regeneration. Scaffolds were fabricated through the powder-metallurgy route utilizing naphthalene as a porogen (porosity = 5-43%). With the inclusion of a higher porogen, a higher fraction of macro-porosity (>20 µm) and pore interconnectivity were observed. X-ray diffraction (XRD) studies confirmed the formation of the farringtonite phase. The developed scaffolds exhibited a minimum ultimate compressive strength (UCS) of 8.5 MPa (for 40 Naph), which lies within the range of UCS of the cancellous bone of humans (2-12 MPa). The in vitro assessment via immersion in physiological fluid yielded a higher deposition of the calcium phosphate (CaP) compound in response to increased macro-porosity and interconnectivity (40 Naph). Cytocompatibility assessed using MC3T3-E1 cells showed that the incorporation of d-PRF coupled with increased porosity resulted the highest cell attachment, proliferation, and viability. For further evaluation, the developed scaffolds were implanted in in vivo rabbit femur condylar defects. Radiography, SEM, OTC labelling, and histology analysis after 2 months of implantation revealed the better invasion of mature osteoblastic cells into the scaffolds with enhanced angiogenesis and superior and accelerated healing of bone defects in d-PRF-incorporated higher porosity scaffolds (40 Naph). Finally, it is hypothesized that the combination of d-PRF incorporation with multiscale porosity and increased interconnectivity facilitated better bone-forming ability, good biocompatibility, and controlled degradability within and around the Zn-doped MgP scaffolds.

Synergistic Improvement of Antibacterial and Osteogenic Differentiation of Thermomechanically Processed Mg-Zr-Sr-Ce Alloy: Insights into the Role of Precipitate Evolution Supported by AIMD Simulation Study.

In the present study, we have discussed the influence of forging temperature (623 K (FT623), 723 K (FT723) and 823 K (FT823)) on microstructure and texture evolution and its implication on mechanical behavior, in vitro-in vivo biocorrosion, antibacterial response, and cytocompatibility of microalloyed Mg-Zr-Sr-Ce alloy. Phase analysis, SEM, and TEM characterization confirm the presence of Mg12Ce precipitate, and its stability was further validated by performing ab initio molecular dynamic simulation study. FT723 exhibits strengthened basal texture, higher fraction of second phases, and particle-stimulated nucleation-assisted DRX grains compared to other two specimens, resulting in superior strength with comparable ductility. FT723 also exhibits superior corrosion resistance mainly due to the strengthened basal texture and lower dislocation density. All the specimens exhibit excellent antibacterial behavior with Gram-negative E. coli, Gram-positive Staphylococcus aureus, and Pseudomonas aeruginosa bacteria. 100% reduction of bacterial growth is observed within 24 h of culture of the specimens. Cytocompatibility was determined by challenging specimen extracts with the MC3T3-E1 cell lines. FT723 specimen exhibits the highest cell proliferation and alkaline phosphatase activity (ALP) because of its superior corrosion resistance. The ability of the specimens to be used in orthopedic implant application was evaluated by in vivo study in rabbit femur. Neither tissue-related infection nor the detrimental effect surrounding the implant was confirmed from histological analysis. Significant higher bone regeneration surrounding the FT723 specimen was observed in SEM analysis and fluorochrome labeling. After 60 days, the FT723 specimen exhibits the highest bone formation, suggesting it is a suitable candidate for orthopedic implant application.

Biomaterials-Based Strategies to Enhance Angiogenesis in Diabetic Wound Healing.

Amidst the present healthcare issues, diabetes is unique as an emerging class of affliction with chronicity in a majority of the population. To check and control its effects, there have been huge turnover and constant development of management strategies, and though a bigger part of the health care area is involved in achieving its control and the related issues such as the effect of diabetes on wound healing and care and many of the works have reached certain successful outcomes, still there is a huge lack in managing it, with maximum effect yet to be attained. Studying pathophysiology and involvement of various treatment options, such as tissue engineering, application of hydrogels, drug delivery methods, and enhancing angiogenesis, are at constantly developing stages either direct or indirect. In this review, we have gathered a wide field of information and different new therapeutic methods and targets for the scientific community, paving the way toward more settled ideas and research advances to cure diabetic wounds and manage their outcomes.

Biomineralized fluorocanasite-reinforced biocomposite scaffolds demonstrate expedited osteointegration of critical-sized bone defects.

The development of patient-specific bone scaffolds that can expedite bone regeneration has been gaining increased attention, especially for critical-sized bone defects or fractures. Precise adaptation of the scaffold to the region of implantation and reduced surgery times are also crucial at clinical scales. To this end, bioactive fluorcanasite glass-ceramic microparticulates were incorporated within a biocompatible photocurable resin matrix following which the biocomposite resin precursor was 3D-printed with digital light processing method to develop the bone scaffold. The printing parameters were optimized based on spot curing investigation, particle size data, and UV-visible spectrophotometry. In vitro cell culture with MG-63 osteosarcoma cell lines and pH study within simulated body fluid demonstrated a noncytotoxic response of the scaffold samples. Further, the in vivo bone regeneration ability of the 3D-printed biocomposite bone scaffolds was investigated by implantation of the scaffold samples in the rabbit femur bone defect model. Enhanced angiogenesis, osteoblastic, and osteoclastic activities were observed at the bone-scaffold interface, while examining through fluorochrome labelling, histology, radiography, field emission scanning electron microscopy, and x-ray microcomputed tomography. Overall, the results demonstrated that the 3D-printed biocomposite bone scaffolds have promising potential for bone loss rehabilitation.

A computational analysis to evaluate deleterious SNPs of GSK3ß, a multifunctional and regulatory protein, for metabolism, wound healing, and migratory processes.

This study focused on GSK-3ß, a critical serine/threonine kinase with diverse cellular functions. However, there is limited understanding of the impact of non-synonymous single nucleotide polymorphisms (nsSNPs) on its structure and function. Through an exhaustive in-silico investigation 12 harmful nsSNPs were predicted from a pool of 172 acquired from the NCBI dbSNP database using 12 established tools that detects deleterious SNPs. Consistently, these nsSNPs were discovered in locations with high levels of conservation. Notably, the three harmful nsSNPs F67C, A83T, and T138I were situated in the active/binding site of GSK-3ß, which may affect the protein's capacity to bind to substrates and other proteins. Molecular dynamics simulations revealed that the F67C and T138I mutants had stable structures, indicating rigidness, whereas the A83T mutant was unstable. Analysis of secondary structures revealed different modifications in all mutant forms, which may affect the stability, functioning, and interactions of the protein. These mutations appear to alter the structural dynamics of GSK-3ß, which may have functional ramifications, such as the formation of novel secondary structures and variations in coil-to-helix transitions. In conclusion, this study illuminates the possible structural and functional ramifications of these GSK-3 nsSNPs, revealing how protein compactness, stiffness, and interactions may affect biological activities.

Enhanced bone regeneration usingAntheraea mylittasilk fibroin and chitosan based scaffold:in-vivoandin-vitrostudy.

In this study, highly interconnected porous scaffolds fromAntheraea mylittasilk fibroin (SF) and chitosan (CH) were fabricated using the freeze-drying method. The weight ratios of SF to CH were varied from 90:10 (SF90/CH10) to 50:50 (SF50/CH50) to prepare the scaffolds from the aqueous suspension of the protein-polysaccharide mix. From the initial optimization of scaffold composition with respect to their microstructure, porosity, and mechanical properties, the SF80/CH20scaffold exhibited the most suitable properties for bone tissue engineering application as compared to others compositions. Hencein-vitrohemocompatibility, protein adsorption, and MG-63 cell culture studies were carried out for SF80/CH20scaffold. The fabricated SF80/CH20scaffold showed a more controlled swelling percentage of 42.8%, with high BSA protein adsorption of 0.39 mg of BSA per gm of the scaffold at 24 h incubation period. Furthermore,in-vitroMG-63 cell culture study onto the fabricated SF80/CH20scaffold elicited excellent MG-63 cell attachment with better biocompatibility and cell viability with increased F-action production from day 3 to day 7 of the cell culture period.In vivobone defect healing in a rabbit tibia model revealed excellent bone healing capacity in SF80/CH20scaffold implanted specimens compared to control ones, as evident from histology and fluorochrome labeling analysis.

Controlled nano Cu precipitation through age treatment: A method to enhance the biodegradation, mechanical, antimicrobial properties and biocompatibility of Fe-20Mn-3Cu alloys.

Iron-manganese (Fe-Mn) based degradable biomaterials have been proven as a suitable substitute to permanent internal fracture-fixation devices. However, lower degradation and bacterial infection are still major concerns. To overcome these limitations, in this work, we have incorporated copper (Cu) in Fe-Mn system. The objective is to produce Cu nano-precipitates and refined microstructure through suitable combination of cold-rolling and age-treatment, so that degradation is improved eventually. High resolution transmission electron microscope (TEM) and scanning transmission electron microscope (STEM) confirmed the Cu rich composition of the nano-precipitates. Number of precipitates increased as aging time increased. Three-dimensional visualization of Fe, Mn and Cu atomic distributions using atom probe tomography (APT), indicated that Cu precipitates were in 15-50 nm range. Large number of nano-precipitates along with lower dislocation density led to highest strength (1078 MPa) and ductility (37 %) for the 6 h age-treated sample. On the other hand, nano-precipitates and refined microstructure resulted highest degradation for the 12 h of age treated sample (0.091 mmpy). When E.Coli bacteria was cultured with the sample extract, significantly higher antibacterial efficacy was observed for the sample having higher nano-precipitates. Higher degradation rate did not cause cyto-toxicity, rather promoted statistically higher cell proliferation (1.5 times within 24 h) in in vitro cell-material interaction studies. In vivo biocompatibility of the alloy containing large nano-precipitates was confirmed from higher new bone regeneration (60%) in rabbit femur model. Overall study suggested that the optimization of the thermo-mechanical processes can effectively tailor the Fe-Mn-Cu alloys for successful internal fracture fixation. STATEMENT OF SIGNIFICANCE: In the present work, we have reported a noble thermo-mechanical approach to simultaneously achieve Cu nano-precipitates and grain refinement in Fe-20Mn-3Cu alloy.

Synergistic Effects of Cerium and Hot Forging on Biodegradation, Antibacterial Properties, and In Vivo Biocompatibility of Microalloyed Mg-Zr-Sr Alloys.

Biodegradable magnesium (Mg)-based alloys are potential candidates for orthopedic applications. In the present study, we have discussed the effect of cerium (Ce) addition and hot forging on mechanical properties, in vitro-in vivo corrosion, antibacterial activity, and cytocompatibility of microalloyed Mg-0.2Zr-0.1Sr-xCe (x = 0 [MZS], 0.5 wt % [MZS-Ce]) alloys. Addition of 0.5 wt % Ce to forged MZS alloys leads to strengthening of the basal texture as well as formation of a higher fraction of dynamic recrystallized (DRX) grains. Hot forging and addition of cerium to the MZS alloy improve both the yield strength and ultimate tensile strength of the forged MZS-Ce alloy by 1.39 and 1.21 times, respectively, compared to those of the forged MZS alloy. The potentiodynamic polarization test in Hank's solution indicates that the corrosion resistance of the forged MZS alloy improves with addition of 0.5 wt % Ce. Uniform distribution of Mg12Ce precipitates, a higher DRX fraction, strengthened texture, and formation of a compact CeO2 passive layer result in 1.68 times reduction in the immersion corrosion rate of the forged MZS-Ce alloy compared to that of the forged MZS alloy. Addition of Ce to the MZS alloy shows excellent antibacterial activity. The forged MZS-Ce alloy exhibited the highest antibacterial efficacy (76.73%). All the alloys show favorable cytocompatibility and alkaline phosphatase (ALP) activity with MC3T3-E1 cells. The improved corrosion resistance of the forged MZS-Ce alloy (95%) leads to higher cell viability compared to that of the forged MZS alloy (85%). In vivo biodegradation and the ability to generate new bones were analyzed by implanting cylindrical samples in the rabbit femur. Histological analysis showed no adverse effects around the implants. Gradual degradation of the implants and higher new bone formation around the forged MZS-Ce sample were confirmed by micro-CT analysis. Bone regeneration around the implants (58.21%) was validated by flurochrome labeling. After 60 days, the forged MZS-Ce alloy showed controlled corrosion and better bone-implant integration, presenting it as a potential candidate for internal fracture fixation materials.

In vitro and in vivo assessment of decellularized platelet-rich fibrin-loaded strontium doped porous magnesium phosphate scaffolds in bone regeneration.

The present work reports the effect of decellularized platelet-rich fibrin (dPRF) loaded strontium (Sr) doped porous magnesium phosphate (MgP) bioceramics on biocompatibility, biodegradability, and bone regeneration. Sustained release of growth factors from dPRF is a major objective here, which conformed to the availability of dPRF on the scaffold surface even after 7 days of in vitro degradation. dPRF-incorporated MgP scaffolds were implanted in the rabbit femoral bone defect and bone rejuvenation was confirmed by radiological examination, histological examination, fluorochrome labeling study, and micro-CT. µ-CT examination of the regained bone samples exhibited that invasion of mature bone in the pores of the MgP2Sr-dPRF sample was higher than the MgP2Sr which indicated better bone maturation capability of this composition. Quantifiable assessment using oxytetracycline labeling showed 73.55 ± 1.12% new osseous tissue regeneration for MgP2Sr-dPRF samples in contrast to 65.47 ± 1.16% for pure MgP2Sr samples, after 3 months of implantation. Histological analysis depicted the presence of abundant osteoblastic and osteoclastic cells in dPRF-loaded Sr-doped MgP samples as compared to other samples. Radiological studies also mimicked similar results in the MgP2Sr-dPRF group with intact periosteal lining and significant bridging callus formation. The present results indicated that dPRF-loaded Sr-doped magnesium phosphate bioceramics have good biocompatibility, bone-forming ability, and suitable biodegradability in bone regeneration.

Copper and cobalt doped bioactive glass-fish dermal collagen electrospun mat triggers key events of diabetic wound healing in full-thickness skin defect model.

The wounds arising out of underlying hyperglycemic conditions such as diabetic foot ulcers demand a multifunctional tissue regeneration approach owing to several deficiencies in the healing mechanisms. Herein, four different types of electrospun microfibers by combining Rohu fish skin-derived collagen (Fcol) with a bioactive glass (BAG)/ion-doped bioactive glass, namely, Fcol/BAG, Fcol/CuBAG, Fcol/CoBAG, and Fcol/CuCoBAG was developed to accelerate wound healing through stimulation of key events such as angiogenesis and ECM re-construction under diabetic conditions. SEM analysis shows the porous and microfibrous architecture, while the EDX mapping provides evidence of the incorporation of dopants inside various inorganic-organic composite mats. The viscoelastic properties of the microfibrous mats as measured by a nano-DMA test show a higher damping factor non-uniform tan-delta value. The maximum ultimate tensile strength and toughness are recorded for fish collagen with copper doped bioactive glass microfibers while the least values are demonstrated by microfibers with cobalt dopant. In vitro results demonstrate excellent cell-cell and cell-material interactions when human dermal fibroblasts (HDFs) were cultured over the microfibers for 48 h. When these mats were applied over full-thickness diabetic wounds in the rabbit model, early wound healing is attained with Fcol/CuBAG, Fcol/CoBAG, and Fcol/CuCoBAG microfibers. Notably, these microfibers-treated wounds demonstrate a significantly (p < 0.01) higher density of blood vessels by CD-31 immunostaining than control, Duoderm, and Fcol/BAG treated wounds. Mature collagen deposition and excellent ECM remodeling are also evident in wounds treated with fish collagen/ion-doped bioactive glass microfibers suggesting their positive role in diabetic wound healing.

Strategies for Enhancing Vascularization of Biomaterial-Based Scaffold in Bone Regeneration.

Despite the recent advances in reconstructive orthopedics; fracture union is a challenge to bone regeneration. Concurrent angiogenesis is a complex process governed by events, delicately entwined with osteogenesis. However, poorly perfused scaffolds have lower success rates; necessitating the need for a better vascular component, which is important for the delivery of nutrients, oxygen, waste elimination, recruitment of cells for optimal bone repair. This review highlights the latest strategies to promote biomaterial-based scaffold vascularization by incorporation of cells, growth factors, inorganic ions, etc. into natural or synthetic polymers, ceramic materials, or composites of organic and inorganic compounds. Furthermore, it emphasizes structural modifications, biophysical stimuli, and natural molecules to fabricate scaffolds aiding the genesis of dense vascularization following their implantation to manifest a compatible regenerative microenvironment without graft rejection.

Waste-derived biomaterials as building blocks in the biomedical field.

Recent developments in the biomedical arena have led to the fabrication of innovative biomaterials by utilizing bioactive molecules obtained from biological wastes released from fruit and beverage processing industries, and fish, meat, and poultry industries. These biological wastes that end up in water bodies as well as in landfills are an affluent source of animal- and plant-derived proteins, bio ceramics and polysaccharides such as collagens, gelatins, chitins, chitosans, eggshell membrane proteins, hydroxyapatites, celluloses, and pectins. These bioactive molecules have been intricately designed into scaffolds and dressing materials by utilizing advanced technologies for drug delivery, tissue engineering, and wound healing relevance. These biomaterials are environment-friendly, biodegradable, and biocompatible, and show excellent tissue regeneration attributes. Additionally, being cost-effective they can reduce the burden on the healthcare system as well as provide a sustainable solution to waste management. In this review, the current trends in the utilization of plant and animal waste-derived biomaterials in various biomedical fields are considered along with a separate section on their applications as xenografts.

Engineering Vascularizing Electrospun Dermal Grafts by Integrating Fish Collagen and Ion-Doped Bioactive Glass.

Utilizing bioactive molecules from organic sources in combination with inorganic materials for enhanced tissue regeneration has been a focus of recent scientific advancements. Some recent studies showed the potential of some specialized bioactive glass for healing of soft tissues; the role of Rohu (Labeo rohita) skin-derived collagen, a biopolymer in tissue regeneration and cutaneous healing, is yet to be established. So, we have fabricated four different types of electrospun mats as wound dressing materials/dermal grafts by combining locally sourced fish (Rohu) skin-derived collagen with novel composition of bioactive glass (Fcol/BAG) without and with dopants (3% and 5% Cu and Co, respectively and their binary) aimed at achieving an accelerated wound healing. FTIR and EDX mapping indicated successful integration of collagen and ion-doped bioactive glass in electrospun mats. Microfibers' architectural features and composition provided a cytocompatible and nontoxic environment conducive to adhesion, spreading, and proliferation of human dermal fibroblasts in vitro; in addition, they were hemocompatible with rabbit red blood cells. Better cutaneous wound healing in rabbits was achieved by treating with Fcol/CoBAG and Fcol/CuCoBAG microfibers with respect to improved wound closure, well-formed continuous epidermis, higher wound maturity, and regulated deposition of extracellular matrix components; mature collagen and elastin. Notably, a significantly (p < 0.01) higher density of blood vessels/positive CD 31 staining was observed in fish collagen/ion-doped bioactive glass microfibrous mat treated wounds suggesting efficient neo-vascularization during early stages of the healing process particularly attributable to copper and cobalt ions in the doped bioactive glass. Enhanced vascularizing ability of these engineered dermal composite grafts/wound dressings along with efficient remodeling of cutaneous structural components (ECM) could collectively be ascribed to bioactive properties of bioactive glass and stimulatory roles of copper, cobalt ions, and fish collagen. Our study demonstrates that a fish collagen/Cu and Co-doped bioactive glass microfibrous mat could potentially be used as a low-cost dressing material/dermal graft for augmented cutaneous wound healing.

Decellularized xenogenic cartilage extracellular matrix (ECM) scaffolds for the reconstruction of osteochondral defects in rabbits.

The use of decellularized native allogenic or xenogenic cartilaginous extracellular matrix (ECM) biomaterials is widely expanding in the fields of tissue engineering and regenerative medicine. In this study, we aimed to develop an acellular, affordable, biodegradable, easily available goat conchal cartilaginous ECM derived scaffolding biomaterial for repair and regeneration of osteochondral defects in rabbits. Cartilages harvested from freshly collected goat ears were decellularized using chemical agents, namely, hypotonic-hypertonic (HH) buffer and Triton X-100 solution, separately. The morphologies and ultrastructure orientations of the decellularized cartilages remained unaltered in spite of complete cellular loss. Furthermore, when the acellular cartilaginous ECMs were cultured with murine mesenchymal stem cells (MSCs) (C3H10T1/2 cells), cellular infiltration and proliferation were thoroughly monitored using SEM, DAPI and FDA stained images, whereas the MTT assay proved the biocompatibility of the matrices. The increasing amounts of secreted ECM proteins (collagen and sGAG) indicated successful chondrogenic differentiation of the MSCs in the presence of the treated cartilage samples. In vivo biocompatibility studies showed no significant immune response or tissue rejection in the treated samples but tissue necrosis in control samples after 3 months. Upon implantation of the constructs in rabbits' osteochondral defects for 3 months, the histological and micro-CT evaluation revealed significant enhancement and regeneration of neocartilage and subchondral bony tissues. The IGF-1 loaded cartilaginous constructs showed comparatively better healing response after 3 months. Our results showed that decellularized xenogenic cartilaginous biomaterials preserved the bioactivity and integrity of the matrices that also favored in vitro stem cell proliferation and chondrogenic differentiation and enabled osteochondral regeneration, thus paving a new way for articular cartilage reconstruction.

Role of calcium phosphate and bioactive glass coating on in vivo bone healing of new Mg-Zn-Ca implant.

Present investigation focuses on development and detailed characterization of a new Mg alloy sample (BM) with and without coating of hydroxyapatite (BMH) and bioactive glass (BMG) by air plasma spray method. After detailed mechano-physico-chemical characterization of powders and coated samples, electrochemical corrosion and SBF immersion tests were carried out. Detailed in vitro characterizations for cell viability were undertaken using MG-63 cell line followed by in vivo tests in rabbit model for studying bone healing up to 60 days. Starting current density increases from BM to BMH to BMG indicating highest resistance towards corrosion in case of BMG samples, however BMH also showed highest icorr value suggesting slowest rate of corrosion than BM and BMG samples. Dissolution of calcium ion in case of BMH and BMG control formation of apatite phases on surface. Ca2+ ions of coatings and from SBF solution underwent reduction reaction simultaneously with conversion of Mg to MgCl2 releasing OH- in the solution, which increases pH. Viability and propagation of human osteoblast-like cells was verified using confocal microscopy observations and from expression of bone specific genes. Alkaline phosphatase assay and ARS staining indicate cell proliferation and production of neo-osseous tissue matrix. In vivo, based on histology of heart, kidney and liver, and immune response of IL-2, IL-6 and TNFα, all the materials show no adverse effects in body system. The bone creation was observed to be more for BMH. Although both BMH and BMG show rays of possibilities in early new bone formation and tough bone-implant bonding at interface as compared to bare Mg alloy, however, BMG showed better well-sprayed coating covering on substrate and resistance against corrosion prior implanting in vivo. Also, better apatite formation on this sample makes it more favourable implant.

Effects of multiscale porosity and pore interconnectivity on in vitro and in vivo degradation and biocompatibility of Fe-Mn-Cu scaffolds.

Iron (Fe) based scaffolds are promising candidates as degradable metallic scaffolds. High strength and ability to control the degradation with tailormade composition and porosity are specific advantages of these scaffolds. In this research work, iron-manganese-copper (Fe-Mn-Cu) based scaffolds, with multiscale porosity, are developed through a powder metallurgy route using naphthalene as a spacer material. The porosity in the scaffolds ranged from 42-76%, where the majority of the macro-pores (≥20 µm) form an interconnected channel network. XRD analysis confirms the presence of MRI compatible and antiferromagnetic austenite as a major phase in all the scaffolds. The developed scaffolds in this study have a minimum ultimate compressive strength of 7.21 MPa (for 30Naph), which lies within the range of the human cancellous bone UCS (2-12 MPa). The degradation rates of the scaffolds are determined from static immersion tests, where the scaffold with the highest porosity (76%) shows a highest degradation rate of 2.71 mmpy when immersed in Hank's balanced salt solution (HBSS) at 37 °C for 30 days. The increased degradation rate of the scaffolds has no cytotoxic effects on MG63 cells as studied by alamar blue assay and live/dead imaging. When implanted in a rabbit femur, the scaffold with higher porosity showed enhanced osteogenesis, as evident through micro-CT and histological analysis. It is hypothesized that the presence of multiscale porosity with a high degree of interconnectivity facilitated better bone regeneration within and around the Fe-Mn-Cu scaffolds.