Decellularized extracellular matrix from bovine ovarian tissue maintains the protein composition of the native matrisome.

Recent approaches of regenerative reproductive medicine investigate the decellularized extracellular matrix to develop a transplantable engineered ovary (TEO). However, a full proteomic analysis is not usually performed after the decellularization process to evaluate the preservation of the extracellular matrix (ECM). In this study, the ECM of the bovine ovarian cortex was analyzed before and after decellularization using mass spectrometry and bioinformatics. A total of 155 matrisome proteins were identified in the native ECM of the bovine ovarian cortex, with 145 matrisome proteins detected in the decellularized ECM. After decellularization, only 10 matrisome proteins were lost, and notably, none belonged to the category of reproductive biological processes. Histology and histochemistry analyses were employed to assess the general morphology of both native and decellularized ECM, allowing for the identification of the most abundant ECM proteins. Moreover, our study highlighted collagen type VI alpha 3 and heparan sulfate proteoglycan 2 as the most abundant components in the bovine ovarian ECM, mirroring the composition observed in the human ovary. These findings enhance our understanding of the composition of both native and decellularized ECM, with the potential implications for the development of a TEO. SIGNIFICANCE: The significance of the present study lies on the possibility of advancing towards developing a bioengineered ovary, which is the ultimate strategy to regain fertility in women. The results demonstrate that the decellularized extracellular matrix of the bovine ovary maintains the protein composition of the native matrisome, using a recently described decellularization protocol. The decellularized matrix may serve as scaffolding for seeding ovarian stromal cells and follicles to create a bioengineered ovary, and as closer its composition is to the native matrix the better. Also, comparing the bovine ovarian matrisome, which was described for the first time here, with the human ovarian matrisome, we could see a great similarity, suggesting that the bovine ovary decellularized matrix may serve as a model for developing a human bioengineered ovary.

Bone Tissue Engineering With Chitosan, Carbon Nanotubes, and Hydroxyapatite Biomaterials Enriched With Mesenchymal Stem Cells: A Radiographic and Histological Evaluation in a Sheep Model Undergoing Ostectomy (Bone Tissue Engineering in a Sheep Model).

Comminuted fractures associated with tissue loss can adversely affect bone regeneration. Biomaterials enriched with mesenchymal stem cells (MSCs) employed for supporting osteosynthesis and potentiating osteoconduction are necessary to fill these bone defects. Natural compound biomaterials, similar to bone tissue, have been extensively tested in animal models for clinical use. Bone tissue engineering studies have used critical-size defects in ovine tibia monitored by imaging and histological examinations to evaluate the regenerative process. This study aimed to monitor the regenerative process in ovine tibial defects with or without chitosan, carbon nanotubes, or hydroxyapatite biomaterials, enriched or not enriched with MSCs. A 3-cm ostectomy was performed in 18 female Suffolk sheep. A 10-hole 4.5 mm narrow locking compression plate was used for osteosynthesis. The animals were randomly divided into three groups (n = 6): control (CON); defects filled with chitosan, carbon nanotubes, and hydroxyapatite biomaterial (BIO); and the same biomaterial enriched with bone marrow MSCs (BIO + CELL). The animals were evaluated monthly using radiographic examinations until 90 postoperative days, when they were euthanized. The limbs were subjected to micro-computed tomography (micro-CT), and bone specimens were subjected to histological evaluations. The radiographic examinations revealed construction stability without plate deviation, fracture, or bone lysis. Micro-CT evaluation demonstrated a difference in bone microarchitecture between the CON and biomaterial treatment groups (BIO and BIO + CELL). In the histological evaluations, the CON group did not demonstrate bone formation, and in the treatment groups (BIO and BIO + CELL), biocompatibility with sheep tissue was noted, and bone formation with trabeculae interspersed with remnants of the biomaterial was observed, with no differences between the groups. In conclusion, biomaterials present osteoconduction with beneficial characteristics for filling bone-lost fractures, and MSCs did not interfere with bone formation.

Biomimetic bilayer scaffold from Bombyx mori silk materials for small diameter vascular applications in tissue engineering.

Enhancing the biocompatibility and mechanical stability of small diameter vascular scaffolds remain significant challenges. To address this challenge, small-diameter tubular structures were electrospun from silk fibroin (SF) from silk textile industry discarded materials to generate bilayer scaffolds that mimic native blood vessels, but derived from a sustainable natural material resource. The inner layer was obtained by directly dissolving SF in formic acid, while the middle layer (SF-M) was achieved through aqueous concentration of the protein. Structural and biological properties of each layer as well as the bilayer were evaluated. The inner layer exhibited nano-scale fiber diameters and 57.9% crystallinity, and degradation rates comparable with the SF-M layer. The middle layer displayed micrometer-scale fibers diameters with an ultimate extension of about 274%. Both layers presented contact angles suitable for cell growth and cytocompatibility, while the bilayer material displayed an intermediate mechanical response and a reduced enzymatic degradation rate when compared to each individual layer. The bilayer material emulates many of the characteristics of native small-diameter vessels, thereby suggesting further studies towards in vivo opportunities.

A preliminary study of cell-based bone tissue engineering into 3D-printed ß-tricalcium phosphate scaffolds and polydioxanone membranes.

Treatment of complex craniofacial deformities is still a challenge for medicine and dentistry because few approach therapies are available on the market that allow rehabilitation using 3D-printed medical devices. Thus, this study aims to create a scaffold with a morphology that simulates bone tissue, able to create a favorable environment for the development and differentiation of osteogenic cells. Moreover, its association with Plenum Guide, through cell-based tissue engineering (ASCs) for guided bone regeneration in critical rat calvarial defects. The manufacturing and characterization of 3D-printed ß-TCP scaffolds for experimental surgery was performed. Nine male rats were divided into three groups: ß-TCP + PDO membrane (TCP/PG), ß-TCP/ASCs + PDO membrane (TCPasc/PG), and ß-TCP/ASCs + PDO membrane/ASCs (TCPasc/PGasc). A surgical defect with a 5-mm diameter was performed in the right parietal bone, and the defect was filled with the 3D-printed ß-TCP scaffold and PDO membrane with or without ASCs. The animals were euthanized 7, 14, and 30 days after the surgical procedure for histomorphometric and immunolabeling analyses. 3D-printed ß-TCP scaffolds were created with a 404 ± 0.0238 µm gyroid macro-pore and, the association to cell-based therapy promotes, especially in the TCPasc/PGasc group, a bone area formation at the defect border region and the center of the defect. The use of 3D-printed ß-TCP scaffolds and PDO membranes associated with cell-based therapy could improve and accelerate guided bone regeneration, promoting an increase in osteogenic capacity and reducing the time involved in the bone formation process. Moreover, using ASCs optimized the bioceramics by increasing its osteoinductive and osteoprogenitor capacity and, even with the resorption of the printed scaffold, aided as a scaffold for mesenchymal cell differentiation, as well as in bone tissue formation.

Cotton cellulose nanofiber/chitosan scaffolds for skin tissue engineering and wound healing applications.

Chitosan (CS) is a promising polymeric biomaterial for use in scaffolds forin vitroskin models and wound dressings, owing to its non-antigenic and antimicrobial properties. However, CS often exhibits insufficient physicochemical properties, mechanical strength, and bioactivity, limiting its efficacy in demanding applications. To address these challenges, cotton cellulose nanofibers (CNFs) represent a promising nanomaterial for enhancing CS-based scaffolds in tissue engineering. CNF offers superior stiffness, and mechanical properties that enhance cellular adhesion and proliferation, both crucial for effective tissue regeneration and healing. This study aimed to develop and characterize a scaffold combining cotton CNF and CS, focusing on its cytocompatibility with human fibroblasts and keratinocytes. The cotton CNF/CS scaffold was fabricated using the casting technique, and its physicochemical properties and cellular compatibility were assessedin vitro. The results demonstrated that incorporating cotton CNF significantly enhanced the stability of the CS matrix. The CS scaffold with 1000 µg ml-1of cotton CNF exhibited increased roughness and reduced rupture strain compared to the pure CS scaffold. The cotton CNF/CS scaffold effectively promoted the adhesion, viability, proliferation, migration, and collagen synthesis of skin cells. Notably, increased cell viability was observed in human fibroblasts cultured on scaffolds with higher concentrations of cotton CNF (100 and 1000 µg ml-1). Based on the findings, the cotton CNF/CS scaffold demonstrates enhanced physicochemical properties and bioactivity, making it a promising candidate for the development ofin vitrohuman skin models and wound healing dressings.

Unveiling the impact of hypodermis on gene expression for advancing bioprinted full-thickness 3D skin models.

3D skin models have been explored as an alternative method to the use of animals in research and development. Usually, human skin equivalents comprise only epidermis or epidermis/dermis layers. Herein, we leverage 3D bioprinting technology to fabricate a full-thickness human skin equivalent with hypodermis (HSEH). The collagen hydrogel-based structure provides a mimetic environment for skin cells to adhere, proliferate and differentiate. The effective incorporation of the hypodermis layer is evidenced by scanning electron microscopy, immunofluorescence, and hematoxylin and eosin staining. The transcriptome results underscore the pivotal role of the hypodermis in orchestrating the genetic expression of a multitude of genes vital for skin functionality, including hydration, development and differentiation. Accordingly, we evidence the paramount significance of full-thickness human skin equivalents with hypodermis layer to provide an accurate in vitro platform for disease modeling and toxicology studies.

Comparing solution blow spinning and electrospinning methods to produce collagen and gelatin ultrathin fibers: A review.

Ultrathin fibers have been used to design functional nanostructured materials for technological and biomedical applications. Combining the use of renewable and compatible sources with the emerging alternative SBS (solution blow spinning) technique opens new opportunities for material applications. In this review, we introduce the benefits of SBS over the classical electrospinning technique by following studies that use collagen or gelatin. SBS offers distinct advantages over electrospinning in the preparation of ultrathin fibers based on natural proteins, including the absence of high-voltage sources and the possibility of using fewer toxic solvents. Notably, there is also the prospect of using SBS directly in injured tissues, opening new strategies for in situ structure assembly SBS is a suitable approach to produce fibers at the nanoscale that can be tailored to distinct diameters by blending or simply adjusting experimental conditions. The focus on producing collagen or gelatin fibers contributes to designing highly biocompatible mats with potential for promoting cellular growth and implantation, even though their applications can be found also in food packaging, energy, and the environment. Therefore, a comprehensive analysis of the topic is essential to evaluate the current strategies regarding these materials and allow for their expanded production and advanced applications.

Optimizing Electroconductive PPy-PCL Scaffolds for Enhanced Tissue Engineering Performance.

The integration of electrically conductive materials is a promising approach in tissue regeneration research. The study presented focuses on the creation of electroconductive scaffolds made from polypyrrole-polycaprolactone (PPy-PCL) using optimal processing parameters. Utilizing Box-Behnken response surface methodology for in situ chemical polymerization of PPy, the scaffolds exhibited a maximum conductivity of 2.542 mS/cm. Morphological examination via scanning electron microscopy (SEM) indicated uniform dispersion of PPy particles within PCL fibers. Fourier transform infrared spectroscopy (FTIR) and energy dispersive x-ray (EDX) analysis validated the composition of the scaffolds, while mechanical testing revealed that the optimized scaffolds exhibit superior tensile strength and Young's modulus compared to scaffolds comprised only of PCL. The hydrophilicity of the scaffolds was improved considerably, transitioning from initially hydrophobic to fully hydrophilic for the optimum scaffold, making it suitable for tissue engineering applications. Cell viability assays, including MTT with L929 fibroblasts and Alamar Blue with bone marrow mesenchymal stem cells (bmMSCs), reflected no cytotoxicity. They showed an increase in metabolic activity, suggesting the capability of the scaffolds to support cellular functions. In conclusion, the in situ synthesis of PPy in the PCL matrix by optimizing the fabrication parameters resulted in conductive scaffolds with promising structural and functional properties for tissue engineering.

The proliferation and viability of human periodontal ligament stem cells cultured on polymeric scaffolds can be improved by low-level laser irradiation.

This study assessed the impact of low-level laser irradiation on the viability and proliferation of human periodontal ligament stem cells (hPDLSCs) cultivated on polylactic acid (PLA) scaffolds. hPDLSCs were obtained, characterized, and grown on the surface of PLA films produced via the solvent casting technique. The study involved two groups: the control group, which was not exposed to radiation, and the laser group, which was irradiated with a diode laser (InGaAIP) with a power of 30 mW, a wavelength of 660 nm, and a single dose of 1 J/cm² emitted continuously. Cell viability was assessed 24 and 48 hours after irradiation using the Alamar blue and Live/Dead assays. Flow cytometry was used to assess cell cycle events, and scanning electron microscopy (SEM) was used to evaluate the interaction between cells and the biomaterial. The results revealed a statistically significant increase in cell metabolic activity in the laser group compared with the control group at 24 hours (p <0.05) and 48 hours (p <0.001), as indicated by the Alamar blue assay. The Live/Dead assay also revealed a greater density of viable cells in the laser group. The cell cycle analysis revealed a significant increase in the number of cells in the proliferative phase (G2/M) in the laser group compared with the control group (p <0.001). The SEM images demonstrated that the irradiated group had a greater concentration of cells while still maintaining their cell shape and projections. This study demonstrated that photobiomodulation can increase the viability and proliferation of periodontal stem cells cultured on PLA scaffolds, suggesting the potential of this protocol for future studies on periodontal tissue engineering.

Multipotent mesenchymal stromal/stem cell-based therapies for acute respiratory distress syndrome: current progress, challenges, and future frontiers.

Acute respiratory distress syndrome (ARDS) is a critical, life-threatening condition marked by severe inflammation and impaired lung function. Mesenchymal stromal/stem cells (MSCs) present a promising therapeutic avenue due to their immunomodulatory, anti-inflammatory, and regenerative capabilities. This review comprehensively evaluates MSC-based strategies for ARDS treatment, including direct administration, tissue engineering, extracellular vesicles (EVs), nanoparticles, natural products, artificial intelligence (AI), gene modification, and MSC preconditioning. Direct MSC administration has demonstrated therapeutic potential but necessitates optimization to overcome challenges related to effective cell delivery, homing, and integration into damaged lung tissue. Tissue engineering methods, such as 3D-printed scaffolds and MSC sheets, enhance MSC survival and functionality within lung tissue. EVs and MSC-derived nanoparticles offer scalable and safer alternatives to cell-based therapies. Likewise, natural products and bioactive compounds derived from plants can augment MSC function and resilience, offering complementary strategies to enhance therapeutic outcomes. In addition, AI technologies could aid in optimizing MSC delivery and dosing, and gene editing tools like CRISPR/Cas9 allow precise modification of MSCs to enhance their therapeutic properties and target specific ARDS mechanisms. Preconditioning MSCs with hypoxia, growth factors, or pharmacological agents further enhances their therapeutic potential. While MSC therapies hold significant promise for ARDS, extensive research and clinical trials are essential to determine optimal protocols and ensure long-term safety and effectiveness.

Alginate-HEMA Hydrogels as Promising Biomaterials for Bone Regeneration: In Vitro and In Vivo Studies.

In the present work, the osteogenic and angiogenic properties of, previously developed, semi-interpenetrated HEMA-EGDMA polymeric networks (sIPN) with and without alginate with application in bone tissue engineering (BTE) were studied. In vitro characterization studies were performed using rat bone marrow progenitor cells (BMPCs), EA.hy926 endothelial cells, and rat vascular smooth muscle cells (VSMCs). Based on the in vitro results of both this work and previous ones, the hydrogels were selected to carry out in vivo studies to find out their capacity as a biomaterial using a bone regeneration model. Our results indicate that the incorporation of alginate into the HEMA-EGDMA polymeric network promotes osteogenic and angiogenic capacity in cell cultures of BMPCs and both EA.hy926 and VSMCs, respectively, and also increases bone formation and vascular structures in in vivo studies, demonstrating its potential use as a biomaterial in BTE.

Cryogenic 3D Printing of GelMA/Graphene Bioinks: Improved Mechanical Strength and Structural Properties for Tissue Engineering.

Purpose: Tissue engineering aims to recreate natural cellular environments to facilitate tissue regeneration. Gelatin methacrylate (GelMA) is widely utilized for its biocompatibility, ability to support cell adhesion and proliferation, and adjustable mechanical characteristics. This study developed a GelMA and graphene bioink platform at concentrations of 1, 1.5, and 2 mg/mL to enhance scaffold properties for tissue engineering applications. Patients and Methods: Graphene was incorporated into GelMA matrices to improve mechanical strength and electrical conductivity of the bioinks. The compressive strength and thermal stability of the resulting GelMA/graphene scaffolds were assessed through DSC analysis and mechanical testing. Cytotoxicity assays were conducted to determine cell survival rates. Cryoprinting at -30°C was employed to preserve scaffold structure and function. The chorioallantoic membrane (CAM) assay was used to evaluate biocompatibility and angiogenic potential. Results: The integration of graphene significantly amplified the compressive strength and thermal stability of GelMA scaffolds. Cytotoxicity assays indicated robust cell survival rates of 90%, confirming the biocompatibility of the developed materials. Cryoprinting effectively preserved scaffold integrity and functionality. The CAM assay validated the biocompatibility and angiogenic potential, demonstrating substantial vascularization upon scaffold implantation onto chick embryo CAM. Conclusion: Integrating graphene into GelMA hydrogels, coupled with low-temperature 3D printing, represents a potent strategy for enhancing scaffold fabrication. The resultant GelMA/graphene scaffolds exhibit superior mechanical properties, biocompatibility, and pro-vascularization capabilities, making them highly suitable for diverse tissue engineering and regenerative medicine applications.

Development and Biocompatibility Assessment of Decellularized Porcine Uterine Extracellular Matrix-Derived Grafts.

Biomaterials derived from biological matrices have been widely investigated due to their great therapeutic potential in regenerative medicine, since they are able to induce cell proliferation, tissue remodeling, and angiogenesis in situ. In this context, highly vascularized and proliferative tissues, such as the uterine wall, present an interesting source to produce acellular matrices that can be used as bioactive materials to induce tissue regeneration. Therefore, this study aimed to establish an optimized protocol to generate decellularized uterine scaffolds (dUT), characterizing their structural, compositional, and biomechanical properties. In addition, in vitro performance and in vivo biocompatibility were also evaluated to verify their potential applications for tissue repair. Results showed that the protocol was efficient to promote cell removal, and dUT general structure and extracellular matrix composition remained preserved compared with native tissue. In addition, the scaffolds were cytocompatible, allowing cell growth and survival. In terms of biocompatibility, the matrices did not induce any signs of immune rejection in vivo in a model of subcutaneous implantation in immunocompetent rats, demonstrating an indication of tissue integration after 30 days of implantation. In summary, these findings suggest that dUT scaffolds could be explored as a biomaterial for regenerative purposes, which is beyond the studies in the reproductive field.

Cell sheet produced from periodontal ligament stem cells activated by PAR1 improves osteogenic differentiation.

Periodontal regeneration is a challenge, and tissue engineering based on periodontal ligament stem cells (PDLSCs) has been shown to be a promising alternative to this process. However, the need for scaffolds has limited the therapeutic use of PDLSCs. In this context, scaffold-free tissue engineering using the cell sheet (CS) technique has been developed as an alternative approach to improve tissue regeneration. Previously, we showed that Protease-activated receptor-1 (PAR1) can regulate PDLSCs. Herein, we evaluate whether PAR1 influences osteogenesis in CSs produced from PDLSCs, without the use of scaffolds. PDLSCs were isolated and immunophenotyped. Then, CSs were obtained by supplementing the culture medium with ascorbic acid (50 µg/mL), and PAR1 was activated through its agonist peptide (100 nM). Scaffold-free 3D CSs were successfully produced from PDLSCs, and they showed higher proliferation potential than isolated PDLSCs. Also, PAR1 activation decreased senescence and improved osteogenic differentiation of CSs by increasing mineralized nodule deposition and alkaline phosphatase concentration; PAR1 also modulated osteogenic markers at the gene and protein levels. We further demonstrated that this effect was regulated by Wnt, TGF-ßI, MEK, p38 MAPK, and FGF/VEGF signaling pathways in PDLSCs (p < 0.05%). Overall, PAR1 activation increased osteogenic activity in CSs, emerging as a promising scaffold-free therapeutic approach for periodontal regeneration.

3D printing of strontium-enriched biphasic calcium phosphate scaffolds for bone regeneration.

Calcium phosphate (CaP) scaffolds doping with therapeutic ions are one of the focuses of recent bone tissue engineering research. Among the therapeutic ions, strontium stands out for its role in bone remodeling. This work reports a simple method to produce Sr-doped 3D-printed CaP scaffolds, using Sr-doping to induce partial phase transformation from ß-tricalcium phosphate (ß-TCP) to hydroxyapatite (HA), resulting in a doped biphasic calcium phosphate (BCP) scaffold. Strontium carbonate (SrCO3) was incorporated in the formulation of the 3D-printing ink, studying ß-TCP:SrO mass ratios of 100:0, 95:5, and 90:10 (named as ß-TCP, ß-TCP/5-Sr, and ß-TCP/10-Sr, respectively). Adding SrCO3 in the 3D-printing ink led to a slight increase in viscosity but did not affect its printability, resulting in scaffolds with a high printing fidelity compared to the computational design. Interestingly, Sr was incorporated into the lattice structure of the scaffolds, forming hydroxyapatite (HA). No residual SrO or SrCO3 were observed in the XRD patterns of any composition, and HA was the majority phase of the ß-TCP/10-Sr scaffolds. The addition of Sr increased the compression strength of the scaffolds, with both ß-TCP/5-Sr and ß-TCP/10-Sr performing better than the ß-TCP. Overall, ß-TCP/5-Sr presented higher mineralized nodules and mechanical strength, while ß-TCP scaffolds presented superior cell viability. The incorporation of SrCO3 in the ink formulation is a viable method to obtain Sr-BCP scaffolds. Thus, this approach could be explored with other CaP scaffolds aiming to optimize their performance and the addition of alternative therapeutic ions.

Enhancing Lung Recellularization with Mesenchymal Stem Cells via Photobiomodulation Therapy: Insights into Cytokine Modulation and Sterilization.

Several lung diseases can cause structural damage, making lung transplantation the only therapeutic option for advanced disease stages. However, the transplantation success rate remains limited. Lung bioengineering using the natural extracellular matrix (ECM) of decellularized lungs is a potential alternative. The use of undifferentiated cells to seed the ECM is practical; however, sterilizing the organ for recellularization is challenging. Photobiomodulation therapy (PBMT) may offer a solution, in which the wavelength is crucial for tissue penetration. This study aimed to explore the potential of optimizing lung recellularization with mesenchymal stem cells using PBMT (660 nm) after sterilization with PBMT (880 nm). The lungs from C57BL/6 mice were decellularized using 1% SDS and sterilized using PBMT (880 nm, 100 mW, 30 s). Recellularization was performed in two groups: (1) recellularized lung and (2) recellularized lung + 660 nm PBMT (660 nm, 100 mW, 30 s). Both were seeded with mesenchymal stem cells from human tooth pulp (DPSc) and incubated for 24 h at 37 °C and 5% CO2 in bioreactor-like conditions with continuous positive airway pressure (CPAP) at 20 cmH2O and 90% O2. The culture medium was analyzed after 24 h. H&E, immunostaining, SEM, and ELISA assays were performed. Viable biological scaffolds were produced, which were free of cell DNA and preserved the glycosaminoglycans; collagens I, III, and IV; fibronectin; laminin; elastin; and the lung structure (SEM). The IL-6 and IL-8 levels were stable during the 24 h culture, but the IFN-γ levels showed significant differences in the recellularized lung and recellularized lung + 660 nm PBMT groups. Greater immunological modulation was observed in the recellularized groups regarding pro-inflammatory cytokines (IL-6, IFN-γ, and IL-8). These findings suggest that PBMT plays a role in cytokine regulation and antimicrobial activity, thus offering promise for enhanced therapeutic strategies in lung bioengineering.

Graphene-enhanced PCL electrospun nanofiber scaffolds for cardiac tissue engineering.

Cardiovascular diseases, particularly myocardial infarction, have significant healthcare challenges due to the limited regenerative capacity of injured heart tissue. Cardiac tissue engineering (CTE) offers a promising approach to repairing myocardial damage using biomaterials that mimic the heart's extracellular matrix. This study investigates the potential of graphene nanopowder (Gnp)-enhanced polycaprolactone (PCL) scaffolds fabricated via electrospinning to improve the properties necessary for effective cardiac repair. This work aimed to analyze scaffolds with varying graphene concentrations (0.5%, 1%, 1.5%, and 2% by weight) to determine their morphological, chemical, mechanical, and biocompatibility characteristics. The results presented that incorporating graphene improves PCL scaffolds' mechanical properties and cellular interactions. The optimal concentration of 1% graphene significantly enhanced mechanical properties and biocompatibility, promoting cell adhesion and proliferation. These findings suggest that Gnp-enhanced PCL scaffolds at this concentration can serve as a potent substrate for CTE providing insights into designing more effective biomaterials for myocardial restoration.

Osteoconductive composite membranes produced by rotary jet spinning bioresorbable PLGA for bone regeneration.

Bone defects and injuries are common, and better solutions are needed for improved regeneration and osseointegration. Bioresorbable membranes hold great potential in bone tissue engineering due to their high surface area and versatility. In this context, polymers such as poly(lactic-co-glycolic acid) (PLGA) can be combined with osteoconductive materials like hydroxyapatite (HA) nanoparticles (NPs) to create membranes with enhanced bioactivity and bone regeneration. Rotary Jet spinning (RJS) is a powerful technique to produce these composite membranes. This study presents an innovative and efficient method to obtain PLGA-HA(NPs) membranes with continuous fibers containing homogeneous HA(NPs) distribution. The membranes demonstrated stable thermal degradation, allowing HA(NPs) quantification. In addition, the PLGA-HA(NPs) presented osteoconductivity, were not cytotoxic, and had high cell adhesion when cultured with pre-osteoblastic cells. These findings demonstrate the potential of RJS to produce PLGA-HA(NPs) membranes for easy and effective application in bone regeneration.

Which detergent is most suitable for the generation of an acellular pancreas bioscaffold?

Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 µm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.

Design and simulation of scaffolds with lattice microstructures for bioprinting bone tissue.

BACKGROUND: Tissue engineering seeks to improve, maintain, or replace the biological functions of damaged organs or tissues with biological substitutes such as the development of scaffolds. In the case of bone tissue, they must have excellent mechanical properties like native bone. OBJECTIVE: In this work, three geometric models were designed for scaffolds with different structure lattices and porosity that could be biomechanically suitable and support cell growth for trabecular bone replacement applications in tissue engineering and regenerative medicine to the proximal femur area. METHODS: Geometries were designed using computer-aided design (CAD) software and evaluated using finite element analysis in compression tests. Three loads were considered according to the daily activity: 1177 N for slow walking, 2060 N for fast walking, and 245.25 N for a person in a bipedal position. All these loads for an adult weight of 75 kg. For each of them, three biomaterials were assigned: two polymers (poly-glycolic acid (PGA) and poly-lactic acid (PLA)) and one mineral (hydroxyapatite (HA)). 54 tests were performed: 27 for each of the tests. RESULTS: The results showed Young's modulus (E) between 1 and 4 GPa. CONCLUSION: If the resultant E is in the range of 0.1 to 5 GPa, the biomaterial is considered an appropriate alternative for the trabecular bone which is the main component of the proximal bone. However, for the models applied in this study, the best option is the poly-lactic acid which will allow absorbing the acting loads.