Tissue Repair Applications Research Articles

Application of Platelet Rich Fibrin in the Regeneration of Intra Oral Defects: A Systematic Review

Objective: To systematically review the randomized clinical trials (RCTs) on the effect of Platelet Rich Fibrin (PRF) concentrate on the tissue regeneration of intra-oral defects.
 Methods: An electronic search was performed in PubMed/Medline and Cochrane Library using relevant keywords until June 2018. RCTs which used PRF concentrate for treatment of intrabony defects, sinus augmentation, furcation involvement, endo-periodontal lesions, gingival recession, and ridge preservation were included in the present review.
 Results: In total, 79 studies that were used PRF either solely or mixed in human trials were included and divided based on the defect type. Most of the studies used PRF to treat intra-bony defects showed that it would improve treatment outcomes. In case of furcation involvement, the application of L-PRF in addition to open flap debridement improved the bone regeneration in grade II mandibular furcation involvement. In case of ridge preservation and sinus floor augmentation, the results were controversial.
 Conclusions: The result of the current systematic review implied that the treatment outcome of PRF application for periodontal and soft tissue repair depends on the treatment strategies and type of the defect. It was shown to PRF application is a practical approach to accelerate and enhance new bone formation in human studies in intrabony defects and furcation involvements. However, further clinical trials for evaluation of other types of intra-oral defects are required.

Photoresponsive Delivery Microcarriers for Tissue Defects Repair.

Intelligent responsive microcarriers have emerged as a promising class of biomaterials for therapeutic delivery and tissue regeneration, since they can respond to external stimuli and release the loaded drugs in an active manner. Among various available stimuli, near‐infrared (NIR) light is particularly attractive because it can penetrate biotic tissues with sufficient intensity and minimal damage. In this work, a kind of photoresponsive delivery microcarriers (PDMs) is developed using microfluidics. The microcarriers consist of NIR‐absorbing graphene oxide, thermosensitive poly(N‐isopropylacrylamide), and biocompatible gelatin methacrylate. Under NIR light, the PDMs exhibit an evident volume shrinkage and effectively trigger the drug release. After the NIR light is switched off, the shrunken microcarriers return to their original size. This reversible process can be stably repeated for many cycles. An in vitro experiment demonstrates that the NIR‐radiated PDMs can actively release vascular endothelial growth factors and improve the tube formation of human umbilical vein endothelial cells. The results from the in vivo experiment also show an obvious photothermal effect and superior therapeutic efficacy of these PDMs in a rat model of tissue defects. These features make the PDMs an excellent drug delivery system and represent a great potential for clinical applications in tissue repair.

Biofunctionalized cellulose paper matrix for cell delivery applications

The present study delineates the preparation, characterization, and application of (3-Aminopropyl)triethoxysilane (APTES)/Caprine liver-derived extracellular matrix (CLECM) coated paper matrix for cell delivery. Here, we exploited positively charged surface of the paper matrix (as imparted by APTES derivatization) to improve the biological responses of the cells. Our results demonstrated that the functionalized paper matrixes favored the adhesion, growth, and proliferation of multiple cell types including normal, transformed, cancerous, and stem cells as compared to the pristine paper matrix. Upon implantation into the mice model, the developed paper matrix supported infiltration of the host cells and vasculature without showing any evidence of significant systemic toxicity. Moreover, the cells cultured on the paper matrix, when delivered to the CAM and mouse models, showed an enhanced vascular network around the substrate, thereby confirming its potential to deliver the cells in vivo. Together, the study confirms that the reported paper-based platform is easy to fabricate, cheap, portable and could efficiently be applied to cell delivery applications for either tissue repair or the development of humanized animal model.

Mesenchymal Stem Cell Migration and Tissue Repair.

Mesenchymal stem cells (MSCs) are multilineage cells with the ability to self-renew and differentiate into a variety of cell types, which play key roles in tissue healing and regenerative medicine. Bone marrow-derived mesenchymal stem cells (BMSCs) are the most frequently used stem cells in cell therapy and tissue engineering. However, it is prerequisite for BMSCs to mobilize from bone marrow and migrate into injured tissues during the healing process, through peripheral circulation. The migration of BMSCs is regulated by mechanical and chemical factors in this trafficking process. In this paper, we review the effects of several main regulatory factors on BMSC migration and its underlying mechanism; discuss two critical roles of BMSCs—namely, directed differentiation and the paracrine function—in tissue repair; and provide insight into the relationship between BMSC migration and tissue repair, which may provide a better guide for clinical applications in tissue repair through the efficient regulation of BMSC migration.

Stem Cell Extracellular Vesicles and their Potential to Contribute to the Repair of Damaged CNS Cells.

Neurological diseases and disorders are leading causes of death and disability worldwide. Many of these pathologies are associated with high levels of neuroinflammation and irreparable tissue damage. As the global burden of these pathologies continues to rise there is a significant need for the development of novel therapeutics. Due to their multipotent properties, stem cells have broad applications for tissue repair; additionally, stem cells have been shown to possess both immunomodulatory and neuroprotective properties. It is now believed that paracrine factors, such as extracellular vesicles (EVs), play a critical role in the functionality associated with stem cells. The diverse biological cargo contained within EVs are proposed to mediate these effects and, to date, the reparative and regenerative effects of stem cell EVs have been demonstrated in a wide range of cell types. While a high potential for their therapeutic use exists, there is a gap of knowledge surrounding their characterization, mechanisms of action, and how they may regulate cells of the CNS. Here, we report the isolation, characterization, and functional assessment of EVs from two sources of human stem cells, mesenchymal stem cells and induced pluripotent stem cells. We demonstrate the ability of these EVs to enhance the processes of cellular migration and angiogenesis, which are critical for both normal cellular development as well as cellular repair. Furthermore, we investigate their reparative effects on damaged cells, specifically those with relevance to the central nervous system. Collectively, our data highlight the similarities and differences among these EV populations and support the view that stem cells EV can be used to repair or partially reverse cellular damage. Graphical Abstract Stem cell-derived Extracellular Vesicles (EVs) for repair of damaged cells. EVs isolated from human induced pluripotent stem cells and mesenchymal stem cells contribute to the partial reversal of phenotypes induced by different sources of cellular damage.

Dissecting the Hierarchy and Lineage of Mesenchymal Stem Cells Using Mouse Genetics as a Step toward Drug Discovery and Regenerative Medicine

The mesenchymal stem cell (MSC) is a type of tissue stem cell. In clinical studies, cultured MSCs have shown important therapeutic effects on diseases via both the reduction of neurological defects and the regulation of immune responses. However, in vivo MSC localization, function, and properties are poorly understood; therefore, the molecular understanding of MSC hierarchy is less advanced compared to hematopoietic stem cell hierarchy. Runt-related transcription factor 2 (Runx2) is an essential transcriptional regulator of osteoblast differentiation from MSCs. Runx2 deficiency in Paired-related homeobox 1 (Prrx1)-derived cells (Runx2Prrx1-/- mice) results in defective intramembranous ossification. Double-positive cells for Prrx1-GFP, and stem cell antigen-1 (Sca1) (Prrx1+Sca1+ cells) in the calvaria, express Runx2 at lower levels, and are more homogeneous and primitive compared with Prrx1+Sca1- cells. Our results suggest that osteoblast differentiation in vivo may begin at the Prrx1+Sca1+ MSC stage, with sequential progression to Prrx1+Sca1- cells, followed by Osterix+Prrx1-Sca1- osteoblast precursors, which eventually form mature α1(I)-collagen+ osteoblasts. This research will enable us to better understand the in vivo molecular biology features of MSCs, leading to their therapeutic applications for tissue repair and regeneration.

Development and characterization of ZnO piezoelectric thin films on polymeric substrates for tissue repair.

Currently available scaffolds for tissue repair have shown very limited success, so many efforts have being put in the development of novel functional materials capable of regulating cell behavior and enhance the tissue healing rate. Piezoelectric materials, as zinc oxide (ZnO), can be a very interesting solution for scaffold development, as they can deliver electrical signals to cells upon mechanical solicitation, allowing the development of suitable microenvironments for tissue repair. This way, it is reported the deposition of ZnO thin films on a polymer by direct current magnetron sputtering, under different conditions, in order to obtain a piezoelectric ZnO thin film with potential for tissue repair applications. The obtained ZnO thin films were characterized in terms of morphology, crystallography, electrical conductivity, transmittance, piezoelectricity, and adhesion quality. The deposition process resulted in uniform films, with a very good adhesion to the substrate. The different deposition conditions influenced the evolution of the crystalline domains and preferential growths and consequently, the electrical properties of the films. One of the conditions resulted in a thin film with a high piezoelectric coefficient and a conductor behavior, being considered the most promising to act as a bioactive coating.

Mesenchymal Stem Cell-Derived Extracellular Vesicles as Therapeutics and as a Drug Delivery Platform.

SummaryMesenchymal stem cells (MSCs) are one of the most easily accessible stem cells that can be obtained from various human tissues. They have raised considerable interests for their potential applications in tissue repair, anti‐cancer therapy, and inflammation suppression. Stem cell‐based therapy was first used to treat muscular dystrophies and has been studied intensively for its efficacy in various disease models, including myocardial infarction, kidney injuries, liver injuries, and cancers. In this review, we summarized the potential mechanisms underlying MSC‐derived EVs therapy as a drug delivery platform. Additionally, based on currently published data, we predicted a potential therapeutic role of cargo proteins shuttled by EVs from MSCs. These data may support the therapeutic strategy of using the MSC‐derived EVs to accelerate this strategy from bench to bedside. stem cells translational medicine 2019;8:880&886

Progress in developing decellularized bioscaffolds for enhancing skin construction.

The skin is the largest organ in the human body, and skin defects are very common. Skin flap transplantation is the best treatment for serious wound defects, and donor site tissues are always sacrificed during this process. Decellularized biomaterials, derived mainly from various nonautologous organs and tissues, have promising applications in tissue engineering and repair of wound defects. To date, decellularized mesothelium, intestine, amniotic membrane, dermis, and skin flaps have been developed and applied for skin coverage in animal models and clinical practice. In this review, we discuss recent advances in decellularized biomaterials for skin substitutes and future perspectives. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1849-1859, 2019.

Self-Powered Well-Aligned P(VDF-TrFE) Piezoelectric Nanofiber Nanogenerator for Modulating an Exact Electrical Stimulation and Enhancing the Proliferation of Preosteoblasts.

Electric potential plays an indispensable role in tissue engineering and wound healing. Piezoelectric nanogenerators based on direct piezoelectric effects can be self-powered energy sources for electrical stimulation and have attracted extensive attention. However, the accuracy of piezoelectric stimuli on piezoelectric polymers membranes in vitro during the dynamic condition is rarely studied. Here, a self-powered tunable electrical stimulation system for assisting the proliferation of preosteoblasts was achieved by well-aligned P(VDF-TrFE) piezoelectric nanofiber membrane (NFM) both as a nanogenerator (NG) and as a scaffold. The effects of electrospinning and different post-treatments (annealing and poling) on the surface wettability, piezoelectric β phase, ferroelectric properties, and sensing performance of NFMs were evaluated here. The polarized P(VDF-TrFE) NFM offered an enhanced piezoelectric value (d31 of 22.88 pC/N) versus pristine P(VDF-TrFE) NFM (d31 of 0.03 pC/N) and exhibited good sensing performance. The maximum voltage and current output of the P(VDF-TrFE) piezoelectric nanofiber NGs reached −1.7 V and 41.5 nA, respectively. An accurate electrical response was obtained in real time under dynamic mechanical stimulation by immobilizing the NGs on the flexible bottom of the culture plate, thereby restoring the real scene of providing electrical stimulation to the cells in vitro. In addition, we simulated the interaction between the piezoelectric nanofiber NG and cells through an equivalent circuit model. To verify the feasibility of P(VDF-TrFE) nanofiber NGs as an exact electrical stimulation, the effects of different outputs of P(VDF-TrFE) nanofiber NGs on cell proliferation in vitro were compared. The study realized a significant enhancement of preosteoblasts proliferation. This work demonstrated the customizability of P(VDF-TrFE) piezoelectric nanofiber NG for self-powered electrical stimulation system application and suggested its significant potential application for tissue repair and regeneration.

Super tough graphene oxide reinforced polyetheretherketone for potential hard tissue repair applications

Biocompatible polyetheretherketone (PEEK) is a favorable material for hard tissue repair due to its similar elastic modulus to that of the human bone. In this work, graphene oxide (GO) reinforced PEEK nanocomposites with different GO loading have been prepared by injection molding. Mechanical testing reveals that the toughness of the reinforced composite varies with the GO loading, with 0.5% GO giving the greatest elongation at break (86.32% greater than pristine PEEK). The underlying toughening mechanism has been attributed to the well-dispersed GO forming π-π* conjugations at the GO/PEEK interface. These conjugations also acted as the nucleation sites for oriented crystallized region in PEEK. As the GO content increases further (e.g > 0.5%), the fillers tend to agglomerate and would disturb the crystallites of PEEK and serve as stress concentration sites, resulting in decreased toughness. The biocompatibility of the composites has been evaluated in vitro, and the results showed that the addition of GO into PEEK favors the adhesion and spreading of bone marrow stromal stem cells, demonstrating the strong potential of our GO reinforced PEEK composites in applications such as hard tissue repair and replacement.

Freeze-cast composite scaffolds prepared from sol-gel derived 58S bioactive glass and polycaprolactone

This work deals with the preparation of freeze-cast scaffolds using sol-gel derived 58S bioactive glass and a hypoeutectic naphthalene-camphor mixture as the starting powder and freezing vehicle, respectively. After the freeze-casting step, samples were air sintered at 1250 °C for 2 h, which led to the crystallization of 58S. The obtained scaffolds were subsequently infiltrated with poly(ε-caprolactone) (PCL), a biodegradable polymer with potential application for bone tissue repair. The prepared materials were examined by helium pycnometry, laser granulometry, scanning electron microscopy (SEM), Archimedes tests, X-ray microtomography (micro-CT), Fourier transform infrared spectroscopy (FTIR), N2 adsorption, X-ray diffraction (XRD), and uniaxial compression tests. Samples cytotoxicity was evaluated by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction (MTT) and LIVE/DEAD assays. Their biocompatibility was also examined after soaking in a simulated body fluid (SBF) solution at 37 °C for up to 14 days. It was observed that the infiltration of PCL into the 58S scaffolds greatly increased their mechanical stability. Moreover, it was shown that these composites displayed a high cell viability (above 70%), which reveals that they did not interfere in the production of osteoblast cells. A hydroxyapatite coating was observed on the samples surface upon soaking in SBF, reinforcing that they are biocompatible materials. As far as we know, this is the first time that freeze-cast scaffolds were obtained using sol-gel derived 58S particles and a naphthalene-camphor mixture. Besides, as the infiltration of PCL into freeze-cast bioactive glass scaffolds improved their mechanical stability without impairing their bioactivity, this is a promising approach to prepare samples for load-bearing applications in bone tissue engineering.

Three-Dimensional Stable Alginate-Nanocellulose Gels for Biomedical Applications: Towards Tunable Mechanical Properties and Cell Growing.

Hydrogels have been studied as promising materials in different biomedical applications such as cell culture in tissue engineering or in wound healing. In this work, we synthesized different nanocellulose-alginate hydrogels containing cellulose nanocrystals, TEMPO-oxidized cellulose nanocrystals (CNCTs), cellulose nanofibers or TEMPO-oxidized cellulose nanofibers (CNFTs). The hydrogels were freeze-dried and named as gels. The nanocelluloses and the gels were characterized by different techniques such as Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and dynamic mechanical thermal analysis (DMTA), while the biological features were characterized by cytotoxicity and cell growth assays. The addition of CNCTs or CNFTs in alginate gels contributed to the formation of porous structure (diameter of pores in the range between 40 and 150 μm). TEMPO-oxidized cellulose nanofibers have proven to play a crucial role in improving the dimensional stability of the samples when compared to the pure alginate gels, mainly after a thermal post-treatment of these gels containing 50 wt % of CNFT, which significantly increased the Ca2+ crosslinking density in the gel structure. The morphological characteristics, the mechanical properties, and the non-cytotoxic behavior of the CNFT-alginate gels improved bioadhesion, growth, and proliferation of the cells onto the gels. Thus, the alginate-nanocellulose gels might find applications in tissue engineering field, as for instance, in tissue repair or wound healing applications.

Carbon-based nanomaterials as an emerging platform for theranostics

Carbon-based nanomaterials emerge as promising platforms for theranostic applications in disease treatment and tissue repair.

Molecular understanding of hierarchy and lineage of mesenchymal stem cells in vivo

Mesenchymal stem cell (MSC) is a type of tissue stem cell. In clinical studies, cultured MSCs have shown important therapeutic effects on diseases via the reduction of neurological defects and regulation of immune responses. However, in vivo MSC localization, function, and properties are poorly understood; therefore, the molecular understanding of MSCs hierarchy is less advanced compared to hematopoietic stem cell hierarchy. To address these issues, we developed a method that enables us to visualize MSCs, manipulate their function, and analyze their molecular biology in vivo. Paired-related homeobox 1 (Prrx1)-positive cells are transiently observed during limb skeletal development in mice. Prrx1-positive cells form heterogeneous populations comprising multiple mesenchymal progenitors with different lineages that are developing into osteoblasts, chondrocytes, adipocytes, fibroblasts, and tendon and ligament cells. Our results suggest that osteoblast differentiation in the calvaria begins at the Prrx1+Sca1+ MSC stage with sequential progression to Prrx1+Sca1- cells, then Osterix+Prrx1-Sca1- osteoblast precursors, which eventually form mature α1(I)-collagen+ osteoblasts. Using Runt-related transcription factor 2 (Runx2) conditional knockout mice, furthermore, we found that the essential period of Runx2 function in intramembranous ossification likely begins at the Prrx1+Sca1+ MSC stage and ends at the Osterix+Prrx1-Sca1- osteoblast precursor stage (before mature the α1(I)-collagen+ osteoblasts appear). This approach will enable us to understand the in vivo molecular biology features of MSCs, leading to their therapeutic applications for tissue repair and regeneration. This development can also contribute to the field of pluripotent stem cell by enabling the transplantation of lineage-restricted mesenchymal progenitors.

Super heat deflection resistance stereocomplex crystallisation of PLA system achieved by selective laser sintering

Polylacite (PLA) is a bioresorbable polymer with potential application for bone tissue repair; however, low heat distortion temperature (HDT) limits its biomedical applications. Due to the low-melting point of PLA, only fused deposition modelling (FDM) was used as a rapid prototyping technology to form PLA products. Here, the work presents a PLA stereocomplex formation which was facilitated by melt blending PLA and poly(D-lactide). The melting point was successfully increased to 219.2°C. Besides, the PLA stereocomplex-based three-dimensional scaffold was formed by selective laser sintering (SLS). The parameters of SLS (energy density, scan speed, scan spacing, preheating temperature etc.) were also optimised by comparing the microstructure. Compared with the FDM, the SLS provides many benefits for fabricating tissue engineering scaffolds due to its rough powder surface. Results show that the PLA stereocomplex was successfully obtained and crystallinity of the stereocomplex was 59.7%.

Synthesis and characterization of ZnAl-NO3(-CO3) layered double hydroxide: A novel structure for intercalation and release of simvastatin

Recently, simvastatin (SIM) has been introduced as a stimulator for bone generation. Its controlled delivery rate is determinant for this purpose. In this work, layered double hydroxide (LDH) based drug delivery systems are introduced for SIM drug for the first time. It was loaded into a series of ZnAl-NO3(CO3) LDHs using two different synthesis processes, namely the direct coprecipitation in the presence of SIM and the ion exchange of nitrate and carbonate containing LDHs in SIM solution. The structure, surface morphology, drug loading, thermostability, and chemical bonding were characterized, and the drug release profile and release kinetic of intercalated LDHs were studied. The characterization results revealed that the intercalation, surface bonding and release behavior of SIM are different for carbonate and nitrate LDHs and moreover depend on the synthesis process. X-ray photoelectron spectroscopy and differential scanning calorimetry studies revealed the stronger chemical bonds between SIM and interlayer surfaces of the nitrate LDHs than carbonate types. From an ion-exchange process, the loaded drug quantity into LDH was 7.9 and 2 wt% for NO3- and CO3-based LDHs, respectively, while was 16.2 and 5.5 wt% for coprecipitation process. The release behavior of SIM from intercalated compounds was well-described with modified Freundlich and Elovich models, and NO3-based LDH system showed sustained drug delivery compared to that of CO3-based LDH with relatively lower drug loading. The drug release rates of 84% and 93% were found for CO3-based LDH compared to that of 35% and 45% for NO3-based counterpart after 58 h. Taken together, the NO3-based system loaded with SIM drug could be considered as a potential drug delivery system for bone tissue repair applications.

Injectable and Spatially Patterned Microporous Annealed Particle (MAP) Hydrogels for Tissue Repair Applications.

Spatially patterned hydrogels are becoming increasingly popular in the field of regenerative medicine and tissue repair because of their ability to guide cell infiltration and migration. However, postfabrication technologies are usually required to spatially pattern a hydrogel, making these hydrogels difficult to translate into the clinic. Here, an injectable spatially patterned hydrogel is reported using hyaluronic acid (HA)‐based particle hydrogels. These particle hydrogels are sequentially loaded into a syringe to form a pattern and, once injected, they maintain the pattern. The applicability of this hydrogel in a wound healing skin model, a subcutaneous implant model, as well as a stroke brain model is examined and distinct patterning in all models tested is shown. This injectable and spatially patterned hydrogel can be used to create physical or biochemical gradients. Further, this design can better match the scaffold properties within the physical location of the tissue (e.g., wound border vs wound center). This allows for better design features within the material that promote repair and regeneration.

Hydrogen Sulfide-Releasing Fibrous Membranes: Potential Patches for Stimulating Human Stem Cells Proliferation and Viability under Oxidative Stress.

The design of biomaterial platforms able to release bioactive molecules is mandatory in tissue repair and regenerative medicine. In this context, electrospinning is a user-friendly, versatile and low-cost technique, able to process different kinds of materials in micro- and nano-fibers with a large surface area-to-volume ratio for an optimal release of gaseous signaling molecules. Recently, the antioxidant and anti-inflammatory properties of the endogenous gasotramsmitter hydrogen sulfide (H2S), as well as its ability to stimulate relevant biochemical processes on the growth of mesenchymal stem cells (MSC), have been investigated. Therefore, in this work, new poly(lactic) acid fibrous membranes (PFM), doped and functionalized with H2S slow-releasing donors extracted from garlic, were synthetized. These innovative H2S-releasing mats were characterized for their morphological, thermal, mechanical, and biological properties. Their antimicrobial activity and effects on the in vitro human cardiac MSC growth, either in the presence or in the absence of oxidative stress, were here assessed. On the basis of the results here presented, these new H2S-releasing PFM could represent promising and low-cost scaffolds or patches for biomedical applications in tissue repair.

Preparation of cellulose nanofiber-reinforced gelatin hydrogel and optimization for 3D printing applications

Gelatin (GEL) obtained from animals is famous for its biocompatibility and biodegradability. However, its poor mechanical properties limits possible applications as bio-inks to fabricate tissue scaffolds through three-dimensional (3D) printing. In this work, a high strength hydrogel based on cellulose nanofibers and GEL (CNF/GEL) was designed for 3D printing. Scanning electron microscopy and breaking strength results indicated that a CNF filling content of 10% was the best content in the CNF/GEL hydrogels. The rheological properties of the samples with different solid contents were investigated, and the 10%-CNF/GEL-5 hydrogel was proposed for 3D printing. Then, a printing strategy with optimal conditions, including a crosslinking procedure for obtaining a 3D scaffold, was proposed. The biocompatibility of G-10%-CNF/GEL-5 was also investigated using CCK-8 and Hoechst 33342/PI double-staining assays. These results confirmed that the 10%-CNF/GEL-5 composite hydrogel has potential to be used as a 3D bio-ink for application in tissue repair.