Tissue Regeneration Applications Research Articles

Preparation and characterization of a jelly fig (Ficus awkeotsang Makino) polysaccharide-based bioactive 3D scaffold for improved vascularization and skin tissue engineering applications

Jelly fig polysaccharides (JFP) were extracted from Ficus awkeotsang Makino achenes. The yield of JFP was approximately 10–15 %. FT-IR spectrum of the extracted JFP confirmed that it was made of low methoxyl pectin (LMP). 3D scaffolds of JFP (JFP scaffold) were fabricated using ionic crosslinking of 2 % (w/v) JFP solution with Ca2+ ions and freeze-drying. The JFP scaffold showed 73.46 ± 1.97 % porosity and a 12-fold swelling capacity. The porous morphology was also observed in SEM micrographs. JFP scaffolds were completely degraded in 14 days when incubated in 1 mg/mL lysozyme solution, compared to the 50 % degradation observed in PBS alone. The antioxidant activity of the JFP and JFP scaffold was approximately 40 %. The hemolytic assay of the JFP scaffold showed <5 % (3.0 ± 0.4) RBC lysis. The cytocompatibility of the JFP scaffold was evaluated using L929 mouse fibroblasts and human dermal fibroblasts (HDF). The in vitro studies using L929 cells showed that the JFP scaffold is cytocompatible. HDF cells cultured in the presence of JFP scaffolds show a higher fold cell viability, proliferation, and migration. Collagen expression and deposition were also studied, and no significant changes occurred with JFP scaffold treatment. In vivo CAM assay showed an increase in the number and thickness of blood vessels by 1.185-fold and 1.19-fold, respectively. These results confirm the angiogenic property of the JFP scaffold. These biocompatible and bioactive properties of the JFP scaffold could be beneficial for tissue engineering and regenerative medicine applications.

Recent advances in production of sustainable and biodegradable polymers from agro-food waste: Applications in tissue engineering and regenerative medicines

Agro-food waste is a rich source of biopolymers such as cellulose, chitin, and starch, which have been shown to possess excellent biocompatibility, biodegradability, and low toxicity. These properties make biopolymers from agro-food waste for its application in tissue engineering and regenerative medicine. Thus, this review highlighted the properties, processing methods, and applications of biopolymers derived from various agro-food waste sources. We also highlight recent advances in the development of biopolymers from agro-food waste and their potential for future tissue engineering and regenerative medicine applications, including drug delivery, wound healing, tissue engineering, biodegradable packaging, excipients, dental applications, diagnostic tools, and medical implants. Additionally, it explores the challenges, prospects, and future directions in this rapidly evolving field. The review showed the evolution of production techniques for transforming agro-food waste into valuable biopolymers. However, these biopolymers serving as the cornerstone in scaffold development and drug delivery systems. With their role in wound dressings, cell encapsulation, and regenerative therapies, biopolymers promote efficient wound healing, cell transplantation, and diverse regenerative treatments. Biopolymers support various regenerative treatments, including cartilage and bone regeneration, nerve repair, and organ transplantation. Overall, this review concluded the potential of biopolymers from agro-food waste as a sustainable and cost-effective solution in tissue engineering and regenerative medicine, offering innovative solutions for medical treatments and promoting the advancement of these fields.

ADVANCING MESSENGER RNA THERAPEUTICS FOR ORTHO-REGENERATION

Messenger RNA (mRNA) is a new class of drug that can be used to express a therapeutic protein and, in contrast to DNA, is safer and inexpensive. Among its advantages, mRNA will immediately begin to express its encoded protein in the cell cytoplasm. The protein will be expressed for a period of time, after which the mRNA is degraded. There is no risk of genetic damage, one of the concerns with plasmid DNA (pDNA) used in traditional gene therapy approaches. Nevertheless, mRNA application in tissue regeneration and regenerative medicine remains limited. In this case, mRNA must overcome its main hurdles: immunogenicity, lack of stability, and intracellular delivery. Research has been done to overcome these limitations, and the future of mRNA seems promising for tissue repair1,2. This keynote talk will address questions including: What are the opportunities for mRNA to improve outcomes in musculoskeletal tissue repair, in particular bone and cartilage? What are the key factors and challenges to expediting this technology to patient treatment (beyond COVID-19 vaccination)?Acknowledgements: E.R.B thanks the cmRNAbone project funded by the European Union's Horizon 2020 research and innovation program under the grant agreement no. 874790 and the NIH R01 AR074395 from NIAMS for funding her mRNA work.

PLATELET-DERIVED EXTRACELLULAR VESICLES PROMOTE STEM CELL TENOGENIC COMMITMENT IN A BIOENGINEERED TENDON 3D MODEL

Worldwide, tendon disorders are one of the main causes of disability that decrease the quality of life of individuals and represent a substantial economic burden on society. Currently, the main therapies used for tendon injuries are not able to restore tendon functionality, and due to tendons' hypovascular and hypocellular nature, they present a reduced healing capacity, which also limits the success of the available therapies. In order to discover new therapies, extracellular vesicles (EVs), key players in cell-cell communication, have been widely explored for tissue engineering and regenerative medicine applications. Thus, the aim of this study is to assess the role of EVs derived from platelets in stem cell tenogenic commitment using a bioengineered tendon in vitro model for potential use as tendon therapeutic agents. Biomimetic platelet-derived EVs were produced by freeze-thaw cycles of platelets and isolation at different centrifugation speed. To recreate the architecture of tendons, a 3D system consisting of electrospun anisotropic nanofiber scaffolds coated with collagen encapsulating human adipose stem cells (hASCs) and different types of platelet-derived EVs, were produced. Then, the influence of the tendon-mimetic constructs and the distinct EVs populations in the hASCs tenogenic differentiation were assessed over culture time. We observed that the hASCs on the nanofibrous tendon scaffolds, show high cytoskeleton anisotropic organization that is characteristic of tenocytes. Moreover, acting as biological cues, platelet-derived EVs boosted hASCs tenogenic commitment, supported by the increased gene expression of tendon-related markers (SCX and TNMD). Additionally, EVs enhanced the deposition of tendon like extracellular matrix (ECM), as evidenced by the increased gene expression of ECM-related markers such as COL1, COL3, DCN, TNC, and MMP-3, which are fundamental for ECM synthesis and degradation balance. Moreover, EVs induced lower collagen matrix contraction on hASCs, which has been related with lower myofibroblast differentiation. Overall, the results revealed that EVs are capable of modulating stem cells' behavior boosting their tenogenic commitment, through the increased expression of healthy tendon cell markers, potentiating ECM deposition and decreasing cell contractility. Therefore, platelet EVs are a promising biochemical tool, worthy to be further explored, as paracrine signaling that might potentiate tendon repair and regeneration.

Cell Sheet Engineering in Periodontology: A Review

Cell sheet engineering has developed its own place for the regeneration of tissues. It focuses on phenomena occurring at the cellular level, whereas regenerative medicine and tissue engineering aim to create or stimulate new tissue for the treatment of disease. In medicine and dentistry, tissue engineering, cell-based therapies, and regenerative medicine have been progressing rapidly for regeneration procedures. It has emerged as a promising strategy for scaffold-free cell-based regenerative medicine therapy by means of transplantable single or multi-layered cell-dense sheets to achieve tissue repair and regeneration. Cell sheet engineering could be one of the procedures for regeneration of periodontal tissues, which can yield better results. In this review, the authors will highlight the principal techniques of cell sheet engineering and its application in tissue regeneration in periodontal therapy.

Red propolis extract associated to platelet-rich plasma and stromal cells with focus in cell therapy and functional tissue regeneration.

The use of platelet-rich plasma (PRP) and adipose-derived stromal cells (ADSC) have been investigated as a form of wound healing enhancement. The objective of this work was to evaluate the association of red propolis (RP) and PRP as inducers of ADSC for application in tissue regeneration. Adipose tissue post-collection and post-cryopreservation was isolated with type II collagenase, characterized by flow cytometry, and differentiated into osteogenic, chondrogenic and adipose cell. The viability of ADSC was evaluated when exposed to different concentrations of RP using the MTT and trypan blue assay. Acridine orange and ethidium bromide (AO/EB) was performed to evaluate cell death events. Horizontal migration methods were investigated in ADSC using autologous and homologous PRP associated with RP (PRP/RP). All assays were processed in triplicate. Flow cytometry and cellular differentiation showed that type II collagenase was effective for isolating ADSC post-collection and post-cryopreservation. RP extracts at concentrations of up to 50 μg.mL-1 presented no cytotoxic effects. Association of PRP and RP at 25 and 50 μg.ml-1 influenced ADSC migration, with total closure on the seventh day after exposition. The results here presented could stimulate proliferation of ADSC cells that may contribute directly or indirectly to the reconstructive process of tissue regeneration.

Investigating the Alkaline Potential of Mineral Trioxide Aggregate Repair Using Selenium Nanoparticles

This study aimed to determine the effect of adding selenium nanoparticles (SeNPs) to mineral trioxide aggregate (MTA HP) concerning alkalinizing potential. Additionally, it examined the set material after SeNPs incorporation using Field Emission Scanning Electron Microscopy with Energy Dispersive X-ray analysis (FE-SEM/EDX) for characterizing the elemental composition and morphological alterations resulting from the integration of SeNPs. Cement samples, both before and after SeNPs incorporation, were examined using FE-SEM/EDX. The pH level was also measured with a pH-meter previously calibrated with solutions of known pH, to evaluate the alkalinizing activity of the integrated substance at different concentrations of nanoparticles: Group 1 (control): 0% w/w SeNPs, Group 2: 0.5% w/w SeNPs, Group 3: 1% w/w SeNPs, Group 4: 1.5% w/w SeNPs and Group 5: 2% w/w SeNPs after 1, 7, 14, and 30 days in distal water. The data were analyzed by one-way ANOVA and Tukey tests (P≤0.05). According to FE-SEM/EDX, the morphological characteristics indicate that SeNPs were successfully dispersed and integrated into the MTA repair matrix. EDX examination validates the presence of Selenium, confirming successful integration. The findings confirmed that the MTAHP showed a high pH level with a discernible reduction in the alkalinizing activity with each incorporated concentration of (SeNPs) that significantly differed from the control group across various periods at (P≤ 0.05). Consequently, the findings indicate that the addition of SeNPs to MTA HP has a notable impact on the pH of the storage solution, leading to a significant decrease in pH values for all concentrations and periods when compared to the control group. The alkalinizing action of MTA HP is highly affected by the incorporated SeNPs, making it more suitable for application in pulpal tissue. This study contributes to our understanding of the morphological alterations and elemental composition of SeNP-incorporated MTA HP, enhancing its potential applications in dental and tissue regeneration.

Tissue adhesives based on chitosan for biomedical applications.

Chitosan bio-adhesives bond strongly with various biological tissues, such as skin, mucosa, and internal organs. Their adhesive ability arises from amino acid and hydroxyl groups in chitosan, facilitating interactions with tissue surfaces through chemical (ionic, covalent, and hydrogen) and physical (chain entanglement) bonding. As non-toxic, biodegradable, and biocompatible materials, chitosan bio-adhesives are a safe option for medical therapies. They are particularly suitable for drug delivery, wound healing, and tissue regeneration. In this review, we address chitosan-based bio-adhesives and the mechanisms associated with them. We also discuss different chitosan composite-based bio-adhesives and their biomedical applications in wound healing, drug delivery, hemostasis, and tissue regeneration. Finally, challenges and future perspectives for the clinical use of chitosan-based bio-adhesives are discussed.

Recent Advances in Whiskers: Properties and Clinical Applications in Dentistry.

Whiskers are nanoscale, high-strength fibrous crystals with a wide range of potential applications in dentistry owing to their unique mechanical, thermal, electrical, and biological properties. They possess high strength, a high modulus of elasticity and good biocompatibility. Hence, adding these crystals to dental composites as reinforcement can considerably improve the mechanical properties and durability of restorations. Additionally, whiskers are involved in inducing the value-added differentiation of osteoblasts, odontogenic osteocytes, and pulp stem cells, and promoting the regeneration of alveolar bone, periodontal tissue, and pulp tissue. They can also enhance the mucosal barrier function, inhibit the proliferation of tumor cells, control inflammation, and aid in cancer prevention. This review comprehensively summarizes the classification, properties, growth mechanisms and preparation methods of whiskers and focuses on their application in dentistry. Due to their unique physicochemical properties, excellent biological properties, and nanoscale characteristics, whiskers show great potential for application in bone, periodontal, and pulp tissue regeneration. Additionally, they can be used to prevent and treat oral cancer and improve medical devices, thus making them a promising new material in dentistry.

Isolation of Human Adipose-Derived Stromal/Stem Cells from Lipoaspirates.

Adipose tissue is an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Adipose-derived stromal/stem cells (ASCs) have been widely used in tissue engineering and cell therapy. In addition, the clinical application of ASCs in the treatment of inflammation and injury has been proven a success. Here, we describe methods from our own laboratory and the literature for the isolation and expansion of Adipose-derived stromal/stem cells (ASCs). We present a large-scale procedure suitable for processing >100mL volumes of lipoaspirate tissue specimens by collagenase digestion, a related procedure suitable for processing adipose tissue aspirates without digestion, and a procedure suitable for intact human adipose tissue, such as buccal fat pads in the maxillofacial region.

From electricity to vitality: the emerging use of piezoelectric materials in tissue regeneration.

The unique ability of piezoelectric materials to generate electricity spontaneously has attracted widespread interest in the medical field. In addition to the ability to convert mechanical stress into electrical energy, piezoelectric materials offer the advantages of high sensitivity, stability, accuracy and low power consumption. Because of these characteristics, they are widely applied in devices such as sensors, controllers and actuators. However, piezoelectric materials also show great potential for the medical manufacturing of artificial organs and for tissue regeneration and repair applications. For example, the use of piezoelectric materials in cochlear implants, cardiac pacemakers and other equipment may help to restore body function. Moreover, recent studies have shown that electrical signals play key roles in promoting tissue regeneration. In this context, the application of electrical signals generated by piezoelectric materials in processes such as bone healing, nerve regeneration and skin repair has become a prospective strategy. By mimicking the natural bioelectrical environment, piezoelectric materials can stimulate cell proliferation, differentiation and connection, thereby accelerating the process of self-repair in the body. However, many challenges remain to be overcome before these concepts can be applied in clinical practice, including material selection, biocompatibility and equipment design. On the basis of the principle of electrical signal regulation, this article reviews the definition, mechanism of action, classification, preparation and current biomedical applications of piezoelectric materials and discusses opportunities and challenges for their future clinical translation.

Engineered extracellular vesicles for tissue repair and regeneration.

Extracellular vesicles (EVs) are heterogeneous membrane-like vesicles secreted by living cells that are involved in many physiological and pathological processes and act as intermediaries of intercellular communication and molecular transfer. Recent studies have shown that EVs from specific sources regulate tissue repair and regeneration by delivering proteins, lipids, and nucleic acids to target cells as signaling molecules. Nanotechnology breakthroughs have facilitated the development and exploration of engineered EVs for tissue repair. Enhancements through gene editing, surface modification, and content modification have further improved their therapeutic efficacy. This review summarizes the potential of EVs in tissue repair and regeneration, their mechanisms of action, and their research progress in regenerative medicine. This review highlights their design logic through typical examples and explores the development prospects of EVs in tissue repair. The aim of this review is to provide new insights into the design of EVs for tissue repair and regeneration applications, thereby expanding their use in regenerative medicine.

Impact of Incorporating Silver Nanoparticles (AgNPs) into Collagen-PU-PEG Hydrogels for Superior Antibacterial Efficacy in Severe Wounds

Wound healing is a complex process often hindered by microbial infections, particularly in severe wounds like diabetic ulcers and burns. Collagen-based hydrogels offer potential solutions but lack mechanical support and antibacterial properties. This study explores the incorporation of silver nanoparticles (AgNPs) into Collagen-Polyethylene Glycol (PEG) hydrogels crosslinked with polyurethane (PU) for enhanced wound healing. The hydrogels, labeled CPEG-Ag, revealed a dense fibrillar structure resulting from an interconnected matrix; they exhibited superabsorbent swelling capacity (up to 2100%), which increased with AgNPs content from 0 to 5 wt.%, with no significant effects on the crosslinking (%). FTIR spectroscopy demonstrated the formation of a semi-interpenetrating polymeric network, chemically linked by urea bonds and physically by hydrogen bond interactions. In vitro cell viability studies showed non-cytotoxic effects on monocytes and fibroblast cells, with the former displaying the best metabolic activity after 48 hours of evaluation, while the latter exhibited high proliferation during the first 24 hours, indicating the safe use of CPEG-Ag hydrogels. The effective antibacterial activity against Gram-negative bacteria, E. coli, was also studied, demonstrating that even low contents of AgNPs are sufficient for significant bacterial inhibition. These results highlight the potential of CPEG-Ag hydrogels for wound dressings, especially in complex wounds, and pave the way for future tissue regeneration applications.

Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera.

Tilapia, a widely farmed aquaculture fish, produces substantial waste, including viscera that contain extracellular matrix (ECM) utilized as a biomaterial for tissue regeneration applications. Extracting ECM from viscera requires a specific decellularization method, as no standardized protocol exists. This study performed three decellularization methods: sonication, orbital shaking at room temperature, and agitation at 4°C, using SDS and TX100 at concentrations of 0.1% and 0.3%. The effectiveness of each method was assessed through H&E staining, dsDNA quantification, and SEM imaging to verify cellular content removal and ECM structure preservation. Additional analyses, including ATR-FTIR, SDS-PAGE, protein quantification, HPLC, and detergent residue tests, were performed to examine functional groups, collagen composition, protein content, amino acid profiles, and detergent residues in the decellularized samples. The results of H&E staining showed a significant reduction in cellular components in all samples, which was confirmed through DNA quantification. Sonication with 0.3% SDS achieved the highest DNA removal rate (96.5 ± 1.1%), while SEM images revealed that agitation at 4°C with 0.3% TX100 better preserved ECM structure. Collagen was present in all samples, as confirmed by ATR-FTIR analysis, which revealed pronounced spectral peaks in the amide I, II, III, A, and B regions. Samples treated with agitation at 4°C using 0.1% SDS exhibited the highest protein content (875 ± 15 µg/mg), whereas those treated with TX100 had lower detergent residue. Overall, the decellularization methods effectively reduced DNA content while preserving ECM structure and components, highlighting the potential of tilapia viscera as bioscaffolds and offering insights into utilizing fish waste for high-value products.

Transforming layered 2D mats into multiphasic 3D nanofiber scaffolds with tailored gradient features for tissue regeneration.

Multiphasic scaffolds with tailored gradient features hold significant promise for tissue regeneration applications. Herein, this work reports the transformation of two-dimensional (2D) layered fiber mats into three dimensional (3D) multiphasic scaffolds using a 'solids-of-revolution' inspired gas-foaming expansion technology. These scaffolds feature precise control over fiber alignment, pore size, and regional structure. Manipulating nanofiber mat layers and Pluronic F127 concentrations allows further customization of pore size and fiber alignment within different scaffold regions. The cellular response to multiphasic scaffolds demonstrates the number of cells migrated and proliferated onto the scaffolds are mainly dependent on the pore size rather than fiber alignment. In vivo subcutaneous implantation of multiphasic scaffolds to rats reveals substantial cell infiltration, neo tissue formation, collagen deposition, and new vessel formation within scaffolds, greatly surpassing the capabilities of traditional nanofiber mats. Histological examination indicates the importance of optimizing pore size and fiber alignment for promotion of cell infiltration and tissue regeneration. Overall, these scaffolds have potential applications in tissue modeling, studying tissue-tissue interactions, interface tissue engineering, and high-throughput screening for optimized tissue regeneration.

Biocompatible 3D Printed MXene Microlattices for Tissue‐Integrated Antibiotic Sensing

Abstract3D continuous mesoscale architectures of nanomaterials possess the potential to revolutionize real‐time electrochemical biosensing through higher active site density and improved accessibility for cell proliferation. Herein, 3D microporous Ti3C2TX MXene biosensors are fabricated to monitor antibiotic release in tissue engineering scaffolds. The Ti3C2TX‐coated 3D electrodes are prepared by conformal MXene deposition on 3D‐printed polymer microlattices. The Ti3C2TX MXene coating facilitates direct electron transfer, leading to the efficient detection of common antibiotics such as gentamicin and vancomycin. The 3D microporous architecture exposes greater electrochemically active MXene surface area, resulting in remarkable sensitivity for detecting gentamicin (10–1 mM) and vancomycin (100–1 mM), 1000 times more sensitive than control electrodes composed of 2D planar films of Ti3C2TX MXene. To characterize the suitability of 3D microporous Ti3C2TX MXene sensors for monitoring drug elution in bone tissue regeneration applications, osteoblast‐like (MG‐63) cells are seeded on the 3D MXene microlattices for 3, 5, and 7 days. Cell proliferation on the 3D microporous MXene is tracked over 7 days, demonstrating its promising biocompatibility and its clinical translation potential. Thus, 3D microporous Ti3C2TX MXene can provide a platform for mediator‐free biosensing, enabling new applications for in vivo monitoring of drug elution.

Carbon nanotube nanocomposite scaffolds: advances in fabrication and applications for tissue regeneration and cancer therapy.

Carbon nanotube (CNT) nanocomposite scaffolds have emerged as highly promising frameworks for tissue engineering research. By leveraging their intrinsic electrical conductivity and valuable mechanical properties, CNTs are commonly dispersed into polymers to create robust, electrically conductive scaffolds that facilitate tissue regeneration and remodeling. This article explores the latest progress and challenges related to CNT dispersion, functionalization, and scaffold printing techniques, including electrospinning and 3D printing. Notably, these CNT scaffolds have demonstrated remarkable positive effects across various cell culture systems, stimulating neuronal growth, promoting cardiomyocyte maturation, and facilitating osteocyte differentiation. These encouraging results have sparked significant interest within the regenerative medicine field, including neural, cardiac, muscle, and bone regenerations. However, addressing the concern of CNT cytotoxicity in these scaffolds remains critical. Consequently, substantial efforts are focused on exploring strategies to minimize cytotoxicity associated with CNT-based scaffolds. Moreover, researchers have also explored the intriguing possibility of utilizing the natural cytotoxic properties of CNTs to selectively target cancer cells, opening up promising avenues for cancer therapy. More research should be conducted on cutting-edge applications of CNT-based scaffolds through phototherapy and electrothermal ablation. Unlike drug delivery systems, these novel methodologies can combine 3D additive manufacturing with the innate physical properties of CNT in response to electromagnetic stimuli to efficiently target localized tumors. Taken together, the unique properties of CNT-based nanocomposite scaffolds position them as promising candidates for revolutionary breakthroughs in both regenerative medicine and cancer treatment. Continued research and innovation in this area hold significant promise for improving healthcare outcomes.

A cell-free, biomimetic hydrogel based on probiotic membrane vesicles ameliorates wound healing

Probiotic bacteria, such as Lactobacilli, have been shown to elicit beneficial effects in various tissue regeneration applications. However, their formulation as living bacteria is challenging, and their therapeutic use as proliferating microorganisms is especially limited in immunocompromised patients.Here, we propose a new therapeutic avenue to circumvent these shortcomings by developing a bacteriomimetic hydrogel based on membrane vesicles (MVs) produced by Lactobacilli. We coupled MVs from Lactobacillus plantarum and Lactobacillus casei, respectively, to the surface of synthetic microparticles, and embedded those bacteriomimetics into a pharmaceutically applicable hydrogel matrix. The wound microenvironment changes during the wound healing process, including adaptions of the pH and changes of the oxygen supply. We thus performed proteomic characterization of the MVs harvested under different culture conditions and identified characteristic proteins related to the biological effect of the probiotics in every culture state. In addition, we highlight a number of unique proteins expressed and sorted into the MVs for every culture condition. Using different in vitro models, we demonstrated that increased cell migration and anti-inflammatory effects of the bacteriomimetic microparticles were dependent on the culture condition of the secreting bacteria. Finally, we demonstrated the bacteriomimetic hydrogel's ability to improve healing in an in vivo mouse full-thickness wound model. Our results create a solid basis for the future application of probiotic-derived vesicles in the treatment of inflammatory dispositions and stimulates the initiation of further preclinical trials.

From Its Nature to Its Function: Marine-Collagen-Based-Biomaterials for Hard Tissue Applications.

Impact statement This review discusses the research done using marine collagens (MCs) on biomaterials for bone, cartilage, and osteochondral tissue regenerative applications with the underlying technologies that enable their development, and explains the methodologies used to characterize MCs highlighting their importance, namely regarding the performance of derived biomaterials, and the inherent properties of such collagens. In the second part, the applicability of MCs as biomaterials for hard tissue applications was studied, focusing on the mostly applied fabrication techniques. In conclusion, this review describes the major challenges to be overcome and the forecast for the upcoming years concerning the use of MCs.

ZnO and Cu/ZnO-modified Magnesium orthopedic implant with improved osteoblast cellular activity: An in-vitro study

Mg-based biomaterials, due to their desirable mechanical integrity, natural bioabsorbability, and osteopromotive characteristics, are promising candidates for both orthopedic applications, such as the surgical fixation of injured musculoskeletal tissues, and cardiovascular applications like coronary artery stents. In the present study, nano- and micro-sized ZnO and Cu/ZnO bioactive particles were used to fabricate Mg-based biocomposites using friction stir processing (FSP). FSP resulted in significant microstructural modification in terms of grain refinement, uniform redistribution of secondary phase particles and homogenous dispersion of bioactive particles. Long-term biodegradation results through H2 evolution method illustrated that biodegradation rate of the bioabsorbable WE43 alloy was significantly reduced by addition of bioactive particles. In addition, fabricated biocomposites with addition of nano-sized particles resulted in lower biodegradation rate in comparison to those with micro-sized particles. Furthermore, uniform dispersion of bioactive particles resulted in superior in-vitro biocompatibility in contact with MC3T3-E1 mouse osteoblast cells. Mg–ZnO–N and Mg–Cu/ZnO–N biocomposites showed improved cell adhesion with elongated and spread morphology of MC3T3-E1 osteoblast cells. Live/dead staining and MTT assay indicated that number of healthy cells and cell viability after 24 h of incubation increased by uniform distribution of bioactive particles. Moreover, fabricated biocomposites with homogenous distribution of Zn2+ and Cu2+ ions showed higher ALP activity. Our findings propose that Mg–ZnO and Mg–Cu/ZnO biocomposites with homogenous dispersion of bioactive particles can be promising candidates for bone tissue regeneration applications.