Numerous Tissue Engineering Applications Research Articles

Bioactive Hydrogels Inspired by Laminin: An Emerging Biomaterial for Tissue Engineering Applications.

Tissue or organ damage due to severe injuries or chronic diseases can adversely affect the quality of life. Current treatments rely on organ or tissue transplantation which has limitations including unavailability of donors, ethical issues, or immune rejection after transplantations. These limitations can be addressed by tissue regeneration which involves the development of bioactive scaffolds closely mimicking the extracellular matrix (ECM). One of the major components of ECM is the laminin protein which supports several tissues associated with important organs. In this direction, peptide-based hydrogels can effectively mimic the essential characteristics of laminin. While several reports have discussed the structure of laminin, the potential of laminin-derived peptide hydrogels as effective biomaterial for tissue engineering applications is yet to be discussed. In this context, the current review focuses on the structure of laminin and its role as an essential ECM protein. Further, the potential of short peptide hydrogels in mimicking the crucial properties of laminin is proposed. The review further highlights the significance of bioactive hydrogels inspired by laminin - in addressing numerous tissue engineering applications including angiogenesis, neural, skeletal muscle, liver, and adipose tissue regeneration along with a brief outlook on the future applications of these laminin-based hydrogels.

Correlation between optical and shielding properties of phosphate glasses with alkaline oxide and their application

In numerous tissue engineering and dental applications, bioactive glasses are utilized. These glasses have unique characteristics that make them attractive candidates for a variety of applications. A new bioactive glass system with the structure of 45P2O5 − 20CaO − 15CaCL2 − 8KF − (10 − x) Li2O − (x) TiO2 was developed in this study, with x = 2, 6, and 8 mol%. For usage in radiation protective applications, it was evaluated. By using an ultraviolet–visible spectrophotometer, we were able to measure the absorbance (Abs) and transmittance (T %) in the range of wavelengths 190–2500 nm. Furthermore, the optical energy gap of the produced glasses was determined. Using the MIKE software, the mass attenuation coefficients (MAC) of the bioactive glasses under investigation were calculated for energies ranging from 15 to 200 keV. The 𝐿𝐿𝐿𝐿𝐿𝐿, 𝑍𝑍𝑒𝑒𝑒𝑒𝑒𝑒, 𝑁𝑁𝑒𝑒𝑒𝑒𝑒𝑒, 𝐻𝐻𝐻𝐻𝐻𝐻, 𝑇𝑇𝑇𝑇𝑇𝑇, 𝑎𝑎𝑎𝑎𝑎𝑎 𝑀𝑀𝑀𝑀𝑀𝑀 (Linear attenuation coefficient, effective atomic number, effective electron density, half value layer, tenth value layer, and mean free path) of the bioactive glasses were calculated. According to the findings, the addition of titanium dioxide (TiO2) as well as the metal oxide such as Li2O to bioactive glasses generates significant differences in the attenuation characteristics of bioactive glasses. The results indicate that the PCKLT3( 𝑇𝑇𝑇𝑇𝑇𝑇2= 8mol%) bioactive-glass sample had the best attenuation among other samples.

Mesenchymal Stem Cell Studies in the Goat Model for Biomedical Research-A Review of the Scientific Literature.

Simple SummaryThis review article aims to compile the works published in the scientific literature, over the last two decades, that use the goat as an animal model in preclinical studies using stem cells, alone or associated with biomaterials, for the treatment of injury or disease in divers organ systems. These preclinical studies are performed prior to human clinical trials for the implementation of new medical or surgical therapies in clinical practice. Thus, it appears that, in the area of tissue engineering and regenerative medicine, the caprine model is particularly used in studies using stem cells in the musculoskeletal system but, although in a more limited way, also in the field of dermatology, ophthalmology, dentistry, pneumology, cardiology, and urology. It appears that the goat represents a particularly useful animal model for studies related to the locomotor system because of its size, and also because they have a more active behavior than sheep, being more similar to the human species in this aspect. Additionally, the goat knee anatomy and the thickness of the cartilage that covers this joint are closer to that of humans than that of other large animal models commonly used in orthopedic research.Mesenchymal stem cells (MSCs) are multipotent cells, defined by their ability to self-renew, while maintaining the capacity to differentiate into different cellular lineages, presumably from their own germinal layer. MSCs therapy is based on its anti-inflammatory, immunomodulatory, and regenerative potential. Firstly, they can differentiate into the target cell type, allowing them to regenerate the damaged area. Secondly, they have a great immunomodulatory capacity through paracrine effects (by secreting several cytokines and growth factors to adjacent cells) and by cell-to-cell contact, leading to vascularization, cellular proliferation in wounded tissues, and reducing inflammation. Currently, MSCs are being widely investigated for numerous tissue engineering and regenerative medicine applications. Appropriate animal models are crucial for the development and evaluation of regenerative medicine-based treatments and eventual treatments for debilitating diseases with the hope of application in upcoming human clinical trials. Here, we summarize the latest research focused on studying the biological and therapeutic potential of MSCs in the goat model, namely in the fields of orthopedics, dermatology, ophthalmology, dentistry, pneumology, cardiology, and urology fields.

Light‐induced hydrogels derived from poly(ethylene glycol) and acrylated methyl ricinoleate as biomaterials

AbstractHydrogels are hydrophilic crosslinked polymer networks that can absorb large amounts of water. They are used as biomaterials in numerous tissue engineering applications. Considering environmental awareness, the synthesis of biomaterials from renewable resources through green fabrication methods is essential. This study produces thermoresponsive hydrogels from a castor oil‐based monomer, acrylated methyl ricinoleate, and poly(ethylene glycol) via an environmentally friendly synthesis method. A photopolymerization technique is used with a very short reaction time. Characterization of the hydrogels is performed using thermogravimetric analysis, scanning electron microscopy, and Fourier‐transform infrared spectroscopy. Swelling and deswelling profiles are subsequently analyzed. A maximum equilibrium swelling degree of 271% is reached within 30 min. In vitro cytotoxicity assays of the hydrogels and the degradation products are performed to evaluate the biocompatibility. The hydrogels are biocompatible because the cell survival of all hydrogel samples and degradation products is greater than 100% and 85%, respectively. Consequently, the thermoresponsive hydrogels made from renewable raw materials in a green process offer interesting platforms for building biomaterials such as actuators for lab‐on‐a‐chip devices, microfluidics, drug delivery systems, preclinical drug screening models, and regenerative medicine.

An Insight of Nanomaterials in Tissue Engineering from Fabrication to Applications.

Tissue engineering is a research domain that deals with the growth of various kinds of tissues with the help of synthetic composites. With the culmination of nanotechnology and bioengineering, tissue engineering has emerged as an exciting domain. Recent literature describes its various applications in biomedical and biological sciences, such as facilitating the growth of tissue and organs, gene delivery, biosensor-based detection, etc. It deals with the development of biomimetics to repair, restore, maintain and amplify or strengthen several biological functions at the level of tissue and organs. Herein, the synthesis of nanocomposites based on polymers, along with their classification as conductive hydrogels and bioscaffolds, is comprehensively discussed. Furthermore, their implementation in numerous tissue engineering and regenerative medicine applications is also described. The limitations of tissue engineering are also discussed here. The present review highlights and summarizes the latest progress in the tissue engineering domain directed at functionalized nanomaterials.

Decellularization of human amniotic membrane using detergent-free methods: Possibilities in tissue engineering

The human amniotic membrane (HAM) is widely used as a natural scaffold in tissue engineering due to its excellent biological characteristics, including anti-microbial, anti-inflammatory, low immunogenicity, and pro-angiogenic properties. This study aimed to develop simple and cost-effective protocols for the decellularization of HAM (d-HAM) using detergent-free methods, i.e., mechanical force (brushing) and physical treatment (heating 45–55 °C). The effectiveness of the methods of interest was compared with a chemical-based approach (EDTA + NaOH + NH4Cl). The prepared d-HAMs were characterized using a series of physico-chemical, mechanical, and biological evaluations. The results from DAPI staining revealed that the chemical method could completely remove epithelial cells from HAM, while the two other approaches only reduced the number of epithelial cells. All three decellularization methods led to a sharp reduction (P < 0.001) in the DNA content of the tissue samples (< 50 ng/mg). Histological evaluations showed the preservation of the d-HAMs’ integrity along with the conservation of collagen and glycosaminoglycans (GAGs). Although the chemical method caused the lowest mechanical deterioration (3.55 MPa in ultimate tensile stress), the mechanical method preserved the highest hydroxyproline levels (3.13 mg/mL). On the other hand, the physical method (heating to 45 and 50 °C) encouraged cell proliferation more than the chemical and mechanical approaches. All of the samples proved to be suitable for cell attachment and could induce cell migration. In conclusion, the present study showed that the use of detergent-free protocols is applicable for the decellularization of HAM, and the obtained tissues may be considered as inexpensive dressings for numerous tissue engineering applications.

Rotary Jet-Spun Polycaprolactone/Hydroxyapatite and Carbon Nanotube Scaffolds Seeded with Bone Marrow Mesenchymal Stem Cells Increase Bone Neoformation.

Clinically, bone tissue replacements and/or bone repair are challenging. Strategies based on well-defined combinations of osteoconductive materials and osteogenic cells are promising to improve bone regeneration but still require improvement. Herein, we combined polycaprolactone (PCL) fibers, carbon nanotubes (CNT), and hydroxyapatite (nHap) nanoparticles to develop the next generation of bone regeneration material. Fibers formed by rotary jet spinning (RJS) instead of traditional electrospinning (ES) with embedded bone marrow mesenchymal stem cells (BMMSCs) showed the best outcomes to repair rat calvarial defects after 6 weeks. To understand this, it was observed that different morphologies were formed depending on the manufacturing method used. RJS fibers presented a particular topography with rough fibers, which allowed for better cellular growth and cell spreading in vitro around and into a three-dimensional (3D) mesh, while fibers made by ES were more smooth and cellular growth was only measured on the 3D mesh surface. The fibers with incorporated nHap/CNT nanoparticles enhanced in vitro cell performance as indicated by more cellular proliferation, alkaline phosphatase activity, proliferation, and deposition of calcium. Greater bone neoformation occurred by combining three characteristics: the presence of nHap and CNT nanoparticles, the topography of the RJS fibers, and the addition of BMMSCs. RJS fibers with nanoparticles and seeded with BMMSCs showed 10 136 mm3 of bone neoformation, meaning a 10-fold increase compared to using RJS only and BMMSCs (0.853 mm3) and a 5-fold increase from using ES only (2054 mm3) after 6 weeks of implantation. Conversely, none of these approaches used individually showed any significant difference for in vivo bone neoformation, suggesting that their combination is essential for optimizing bone formation. In summary, our work generated a potential material composed of well-defined combinations of suitable scaffolds seeded with BMMSCs for enhancing numerous orthopedic tissue engineering applications.

Fabrication and characterization of biaxially electrospun collagen/alginate nanofibers, improved with Rhodotorula mucilaginosa sp. GUMS16 produced exopolysaccharides for wound healing applications

Fabrication of scaffolds with enhanced mechanical properties and desirable cellular compatibility is critical for numerous tissue engineering applications. This study was aimed at fabrication and characterization of a nanofiber skin substitute composed of collagen (Col)/sodium alginate (SA)/ polyethylene oxide (PEO)/Rhodotorula mucilaginosa sp. GUMS16 produced exopolysaccharides (EPS) were prepared using the biaxial electrospinning technique. This study used collagen extracted from the bovine tendon as a natural scaffold, sodium alginate as an absorber of excess wound fluids, and GUMS16 produced exopolysaccharides as an antioxidant. Collagen was characterized using FTIR and EDS analyses. The cross-linked nanofibers were characterized by SEM, FTIR, tensile, contact-angle, swelling test, MTT, and cell attachment techniques. The average diameter of Col nanofiber was 910 ± 89 nm. The Col and Col-SA/PEO non-woven mats' water contact angle measurement was 41.6o and 56.4o, Col/EPS1%, Col/EPS2%, Col-SA/PEO + EPS1%, and Col-SA/PEO + EPS2% were 61.4o, 58.3o, 38.5o, and 50.6o, respectively. Cell viability of more than 100% was shown in Col-SA/PEO + EPS nanofibers. Also, SEM images of cells on nanofiber scaffolds demonstrated that all nanofibers incorporated with GUMS16-produced EPS have good cell growth and proliferation. The acquired results expressed that the GUMS16-produced EPS can be considered a novel biomacromolecule in electrospun fibers that increase cell viability and proliferation.

Mesoporous Silica Based Nanostructures for Bone Tissue Regeneration

Porous nano-scaffolds provide for better opportunities to restore, maintain, and improve functions of damaged tissues and organs by facilitating tissue regeneration. Various nanohybrids composed of mesoporous silica nanoparticles (MSNs) are being widely explored for tissue engineering. Since biological activity is enhanced by several orders of magnitude in multicomponent scaffolds, remarkable progress has been observed in this field, which has aimed to develop the controlled synthesis of multifunctional MSNs with tuneable pore size, efficient delivering capacity of bioactive factors, as well as enhanced biocompatibility and biodegradability. In this review, we aim to provide a broad survey of the synthesis of multifunctional MSN based nanostructures with exotic shapes and sizes. Further, their promise as a novel nanomedicine is also elaborated with respect to their role in bone tissue engineering. Also, recent progress in surface modification and functionalization with various polymers like poly (l-lactic acid)/poly (ε-caprolactone), polylysine-modified polyethylenimine, poly (lactic-co-glycolic acid), and poly (citrate-siloxane) and biological polymers like alginate, chitosan, and gelatine are also covered. Several attempts for conjugating drugs like dexamethasone and β–estradiol, antibiotics like vancomycin and levofloxaci, and imaging agents like fluorescein isothiocyanate and gadolinium, on the surface modified MSNs are also covered. Finally, the scope of developing orthopaedic implants and potential trends in 3D bioprinting applications of MSNs are also discussed. Hence, MSNs based nanomaterials may serve as improved candidate biotemplates or scaffolds for numerous bone tissue engineering, drug delivery and imaging applications deserving our full attention now.

Multifunctional Fibroblasts Enhanced via Thermal and Freeze-Drying Post-treatments of Aligned Electrospun Nanofiber Membranes

Parallel fibrous scaffolds play a critical role in controlling the morphology of cells to be more natural and biologically inspired. Among popular tissue engineering materials, poly(2-hydroxyethyl methacrylate) (pHEMA) has been widely investigated in conventional forms due to its biocompatibility, low toxicity, and hydrophilicity. However, the swelling of pHEMA in water remains a major concern. To address this issue, randomly oriented and aligned as-spun pHEMA nanofibrous scaffolds were first fabricated at speeds of 300 and 2000 rpm in this study, which were then post-treated using either a thermal or a freeze-drying method. In cell assays, human dermal fibroblasts (HDFs) adhered to the freeze-drying treated substrates at a significantly faster rate, whereas they had a higher cell growth rate on thermally-treated substrates. Results indicated that the structural properties of pHEMA nanofibrous scaffolds and subsequent cellular behaviors were largely dependent on post-treatment methods. Moreover, this study suggests that aligned pHEMA nanofibrous substrates tended to induce regular fibroblast orientation and unidirectionally oriented actin cytoskeletons over random pHEMA nanofibrous substrates. Such information has predictive power and provides insights into promising post-treatment methods for improving the properties of aligned pHEMA scaffolds for numerous tissue engineering applications.

Chitin nanocrystals assisted 3D printing of polycitrate thermoset bioelastomers

Citrate-based thermoset bioelastomer has numerous tissue engineering applications. However, its insoluble and unmeltable features restricted processing techniques for fabricating complex scaffolds. Herein, direct ink writing (DIW) was explored for 3D printing of poly(1, 8-octanediol-co-Pluronic F127 citrate) (POFC) bioelastomer scaffolds considering that POFC prepolymer (pre-POFC) was waterborne and could form a stable emulsion. The pre-POFC emulsion couldn’t be printed, however, chitin nanocrystal (ChiNC) could be as a rheological modifier to tune the flow behavior of pre-POFC emulsion, and thus DIW printing of POFC scaffolds was successfully realized; moreover, ChiNC was also as a supporting agent to prevent collapse of filaments during thermocuring, and simultaneously as a biobased nanofiller to reinforce scaffolds. The rheological analyses showed the pre-POFC/ChiNC inks fulfilled the requirements for DIW printing. The printed scaffolds exhibited low swelling, and good performances in strength and resilence. Furthermore, the entire process was easily performed and eco-friendly.

Low-cost hybrid scaffolds based on polyurethane and gelatin

The production of scaffolds using a combination of synthetic and natural polymers has been widely studied for numerous tissue engineering applications, as it results in a material with superior properties, combining availability, processability, and the strength and resilience of synthetic polymers with the high biocompatibility of natural polymers. In the present study, fibrous membranes composed of polyurethane and gelatin were fabricated by rotary jet spinning and were posteriorly characterized for their morphological, chemical composition, thermal stability, hydrophilic properties as well as cell viability. Viscosity measurements were taken to achieve the critical concentration of the polymeric solution (9% wt/v), and the production of fibers at different rotational speeds (3000, 6000, 9000 and 12,000rpm) was performed to evaluate the effect of rotational speed on fiber diameter and morphology, as observed in scanning electron microscopy analyses. Continuous and bead-free fibers were achieved at 6000rpm with average diameter of 12.5μm. Chemical composition characterization showed the characteristic peaks of both polymers and the absence of the organic solvent, while the addition of gelatin did not affect the thermal stability of the membrane (up to 314°C). Additionally, the water contact angle proved the membrane hydrophilic nature (81.3°). Cell viability assays exhibited cytocompatibility with endothelial cells for 24, 48 and 72h. The results demonstrate that the PU–Gel combination with the rotary jet spinning process is promising to obtain low-cost scaffolds with interesting properties for numerous tissue engineering applications, and, thus, should be further studied.

Transfection of autologous host cells in vivo using gene activated collagen scaffolds incorporating star-polypeptides

It is increasingly being recognised within the field of tissue engineering that the regenerative capacity of biomaterial scaffolds can be augmented via the incorporation of gene therapeutics. However, the field still lacks a biocompatible gene delivery vector which is capable of functionalizing scaffolds for tailored nucleic acid delivery. Herein, we describe a versatile, collagen based, gene-activated scaffold platform which can transfect autologous host cells in vivo via incorporation of star-shaped poly(˪-lysine) polypeptides (star-PLLs) and a plasmid DNA (pDNA) cargo. Two star-PLL vectors with varying number and length of poly(˪-lysine) arms were assessed. In vitro, the functionalization of a range of collagen based scaffolds containing either glycosaminoglycans (chondroitin sulfate or hyaluronic acid) or ceramics (hydroxyapatite or nano-hydroxyapatite) with star-PLL-pDNA nanomedicines facilitated prolonged, non-toxic transgene expression by mesenchymal stem cells (MSCs). We demonstrate that the star-PLL structure confers enhanced spatiotemporal control of nanomedicine release from functionalized scaffolds over a 28-day period compared to naked pDNA. Furthermore, we identify a star-PLL composition with 64 poly(˪-lysine) arms and 5 (˪-lysine) subunits per arm as a particularly effective vector, capable of facilitating a 2-fold increase in reporter transgene expression compared to the widely used vector polyethylenimine (PEI), a 44-fold increase compared to a 32 poly(˪-lysine) armed star-PLL and a 130-fold increase compared to its linear analogue, linear poly(˪-lysine) (L-PLL) from a collagen-chondroitin sulfate gene activated scaffold. In an in vivo subcutaneous implant model, star-PLL-pDNA gene activated scaffolds which were implanted cell-free exhibited extensive infiltration of autologous host cells, nanomedicine retention within the implanted construct and successful host cell transfection at the very early time point of just seven days. Overall, this article illustrates for the first time the significant ability of the star-PLL polymeric structure to transfect autologous host cells in vivo from an implanted biomaterial scaffold thereby forming a versatile platform with potential in numerous tissue engineering applications.

Wide-ranging diameter scale of random and highly aligned PCL fibers electrospun using controlled working parameters

The prominent success of polycaprolactone (PCL) bioengineered fibrous scaffolds has expanded the use of PCL over other polymers in the wide electrospinning literature. Several highly toxic solvent systems providing a good PCL solubility-spinability are recurrently applied in the overwhelming majority of studies. One of the current major focuses revolves around the challenging generation of aligned fibers that are very desirable in numerous tissue engineering applications. Moreover, the critical influence of fiber size on cellular performance has led to the use of different solvent systems for the production of specific nano- or micro-sized PCL fibers while neglecting the solvents toxicity. Therefore, our goal is to use the unconventional solvent system acetic acid/formic acid of very low toxicity, recently defined as the system producing ultra-thin PCL fibers, in a trial to outspread the size range and to tackle fiber alignment. After profound analysis of the effect of varying collector motion and collector design on fiber alignment, a novel collector producing highly aligned PCL fibers based on synchronic mechanical and electrical effects is designed. In a subsequent step, the fiber diameter is manipulated by analyzing 3 influential parameters: polymer concentration, tip-to-collector distance and the frequently overlooked parameter humidity. The parameters fine-tuning study has resulted in very broad PCL fiber diameter ranges of 94–1548 nm and 114–1408 nm for random and aligned fibers respectively. PCL fibers properties are also characterized in this study by means of water contact angle (WCA), X-ray photoelectron spectroscopy (XPS) and tensile measurements. Despite the detection of the same surface chemical composition on all fiber conditions, the WCA values tend to decrease with increased fiber diameters, especially for aligned fibers. Moreover, aligning the fibers and decreasing their size is found to lead to considerably enhanced mechanical properties. Overall, it can be concluded that this work represents a picture-perfect reference for the electrospinning of PCL fibers having different orientations, diameters and physico-chemical properties from a benign solvent system.

A review of fibrin and fibrin composites for bone tissue engineering.

Tissue engineering has emerged as a new treatment approach for bone repair and regeneration seeking to address limitations associated with current therapies, such as autologous bone grafting. While many bone tissue engineering approaches have traditionally focused on synthetic materials (such as polymers or hydrogels), there has been a lot of excitement surrounding the use of natural materials due to their biologically inspired properties. Fibrin is a natural scaffold formed following tissue injury that initiates hemostasis and provides the initial matrix useful for cell adhesion, migration, proliferation, and differentiation. Fibrin has captured the interest of bone tissue engineers due to its excellent biocompatibility, controllable biodegradability, and ability to deliver cells and biomolecules. Fibrin is particularly appealing because its precursors, fibrinogen, and thrombin, which can be derived from the patient’s own blood, enable the fabrication of completely autologous scaffolds. In this article, we highlight the unique properties of fibrin as a scaffolding material to treat bone defects. Moreover, we emphasize its role in bone tissue engineering nanocomposites where approaches further emulate the natural nanostructured features of bone when using fibrin and other nanomaterials. We also review the preparation methods of fibrin glue and then discuss a wide range of fibrin applications in bone tissue engineering. These include the delivery of cells and/or biomolecules to a defect site, distributing cells, and/or growth factors throughout other pre-formed scaffolds and enhancing the physical as well as biological properties of other biomaterials. Thoughts on the future direction of fibrin research for bone tissue engineering are also presented. In the future, the development of fibrin precursors as recombinant proteins will solve problems associated with using multiple or single-donor fibrin glue, and the combination of nanomaterials that allow for the incorporation of biomolecules with fibrin will significantly improve the efficacy of fibrin for numerous bone tissue engineering applications.

Non-invasive tracking of injected bone marrow mononuclear cells to injury and implanted biomaterials.

Tracking the dynamic behaviour of transplanted bone-marrow mononuclear cells (BM-MNCs) is a long-standing research goal. Conventional methods involving contrast and tracer agents interfere with cellular function while also yielding false signals. The use of bioluminescence addresses these shortcomings while allowing for real-time non-invasive tracking in vivo. Given the failures of transplanted BM-MNCs to engraft into injured tissue, biomaterial scaffolds capable of attracting and enhancing BM-MNC engraftment at sites of injury are highly sought in numerous tissue engineering applications. To this end, the results from this study demonstrate a new longitudinal tracking model that can non-invasively determine exogenous BM-MNC homing and engraftment to biomaterials, providing a valuable tool to inform the design of scaffolds with implications for countless tissue engineering applications.

Fundamental insight into the effect of carbodiimide crosslinking on cellular recognition of collagen-based scaffolds.

Carbodiimide stabilised collagen is employed extensively for the fabrication of biologically active materials. Despite this common usage, the effect of carbodiimide crosslinking on cell-collagen interactions is unclear. Here we have found that carbodiimide crosslinking of collagen inhibits native-like, whilst increasing non-native like, cellular interactions. We propose a mechanistic model in which carbodiimide modifies the carboxylic acid groups on collagen that are essential for cell binding. As such we feel that this research provides a crucial, long awaited, insight into the bioactivity of carbodiimide crosslinked collagen. Through the ubiquitous use of collagen as a cellular substrate we feel that this is fundamental to a wide range of research activity with high impact across a broad range of disciplines.

Bio-based polyurethane for tissue engineering applications: How hydroxyapatite nanoparticles influence the structure, thermal and biological behavior of polyurethane composites

In this work, thermoset polyurethane composites were prepared by the addition of hydroxyapatite nanoparticles using the reactants polyol polyether and an aliphatic diisocyanate. The polyol employed in this study was extracted from the Euterpe oleracea Mart. seeds from the Amazon Region of Brazil. The influence of hydroxyapatite nanoparticles on the structure and morphology of the composites was studied using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), the structure was evaluated by Fourier transform infrared spectroscopy (FT-IR), thermal properties were analyzed by thermogravimetry analysis (TGA), and biological properties were studied by in vitro and in vivo studies. It was found that the addition of HA nanoparticles promoted fibroblast adhesion while in vivo investigations with histology confirmed that the composites promoted connective tissue adherence and did not induce inflammation. In this manner, this study supports the further investigation of bio-based, polyurethane/hydroxyapatite composites as biocompatible scaffolds for numerous tissue engineering applications.

Real-Time Bioluminescence Imaging of Cell Distribution, Growth, and Differentiation in a Three-Dimensional Scaffold Under Interstitial Perfusion for Tissue Engineering.

Bioreactor systems allow safe and reproducible production of tissue constructs and functional analysis of cell behavior in biomaterials. However, current procedures for the analysis of tissue generated in biomaterials are destructive. We describe a transparent perfusion system that allows real-time bioluminescence imaging of luciferase expressing cells seeded in scaffolds for the study of cell-biomaterial interactions and bioreactor performance. A prototype provided with a poly(lactic) acid scaffold was used for "proof of principle" studies to monitor cell survival in the scaffold (up to 22 days). Moreover, using cells expressing a luciferase reporter under the control of inducible tissue-specific promoters, it was possible to monitor changes in gene expression resulting from hypoxic state and endothelial cell differentiation. This system should be useful in numerous tissue engineering applications, the optimization of bioreactor operation conditions, and the analysis of cell behavior in three-dimensional scaffolds.

Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells

Tissue engineering strategies for repairing and regenerating articular cartilage face critical challenges to recapitulate the dynamic and complex biochemical microenvironment of native tissues. One approach to mimic the biochemical complexity of articular cartilage is through the use of recombinant bacterial collagens as they provide a well–defined biological ‘blank template’ that can be modified to incorporate bioactive and biodegradable peptide sequences within a precisely defined three–dimensional system. We customized the backbone of a Streptococcal collagen–like 2 (Scl2) protein with heparin–binding, integrin–binding, and hyaluronic acid–binding peptide sequences previously shown to modulate chondrogenesis and then cross–linked the recombinant Scl2 protein with a combination of matrix metalloproteinase 7 (MMP7)– and aggrecanase (ADAMTS4)–cleavable peptides at varying ratios to form biodegradable hydrogels with degradation characteristics matching the temporal expression pattern of these enzymes in human mesenchymal stem cells (hMSCs) during chondrogenesis. hMSCs encapsulated within the hydrogels cross–linked with both degradable peptides exhibited enhanced chondrogenic characteristics as demonstrated by gene expression and extracellular matrix deposition compared to the hydrogels cross–linked with a single peptide. Additionally, these combined peptide hydrogels displayed increased MMP7 and ADAMTS4 activities and yet increased compression moduli after 6 weeks, suggesting a positive correlation between the degradation of the hydrogels and the accumulation of matrix by hMSCs undergoing chondrogenesis. Our results suggest that including dual degradation motifs designed to respond to enzymatic activity of hMSCs going through chondrogenic differentiation led to improvements in chondrogenesis. Our hydrogel system demonstrates a bimodal enzymatically degradable biological platform that can mimic native cellular processes in a temporal manner. As such, this novel collagen–mimetic protein, cross–linked via multiple enzymatically degradable peptides, provides a highly adaptable and well defined platform to recapitulate a high degree of biological complexity, which could be applicable to numerous tissue engineering and regenerative medicine applications.