Navigating the Immunological Crossroads: Mesenchymal Stem/Stromal Cells as Architects of Inflammatory Harmony in Tissue-Engineered Constructs.

In the dynamic landscape of tissue engineering, the integration of tissue-engineered constructs (TECs) faces a dual challenge-initiating beneficial inflammation for regeneration while avoiding the perils of prolonged immune activation. As TECs encounter the immediate reaction of the immune system upon implantation, the unique immunomodulatory properties of mesenchymal stem/stromal cells (MSCs) emerge as key navigators. Harnessing the paracrine effects of MSCs, researchers aim to craft a localized microenvironment that not only enhances TEC integration but also holds therapeutic promise for inflammatory-driven pathologies. This review unravels the latest advancements, applications, obstacles, and future prospects surrounding the strategic alliance between MSCs and TECs, shedding light on the immunological symphony that guides the course of regenerative medicine.

Biological Macromolecule-Based Scaffolds for Urethra Reconstruction.

Urethral reconstruction strategies are limited with many associated drawbacks. In this context, the main challenge is the unavailability of a suitable tissue that can endure urine exposure. However, most of the used tissues in clinical practices are non-specialized grafts that finally fail to prevent urine leakage. Tissue engineering has offered novel solutions to address this dilemma. In this technology, scaffolding biomaterials characteristics are of prime importance. Biological macromolecules are naturally derived polymers that have been extensively studied for various tissue engineering applications. This review discusses the recent advances, applications, and challenges of biological macromolecule-based scaffolds in urethral reconstruction.

Extracellular Matrix-Based and Electrospun Scaffolding Systems for Vaginal Reconstruction.

Congenital vaginal anomalies and pelvic organ prolapse affect different age groups of women and both have significant negative impacts on patients' psychological well-being and quality of life. While surgical and non-surgical treatments are available for vaginal defects, their efficacy is limited, and they often result in long-term complications. Therefore, alternative treatment options are urgently needed. Fortunately, tissue-engineered scaffolds are promising new treatment modalities that provide an extracellular matrix (ECM)-like environment for vaginal cells to adhere, secrete ECM, and be remodeled by host cells. To this end, ECM-based scaffolds or the constructs that resemble ECM, generated by self-assembly, decellularization, or electrospinning techniques, have gained attention from both clinicians and researchers. These biomimetic scaffolds are highly similar to the native vaginal ECM and have great potential for clinical translation. This review article aims to discuss recent applications, challenges, and future perspectives of these scaffolds in vaginal reconstruction or repair strategies.

Prospects and Challenges of Electrospun Cell and Drug Delivery Vehicles to Correct Urethral Stricture.

Current therapeutic modalities to treat urethral strictures are associated with several challenges and shortcomings. Therefore, significant strides have been made to develop strategies with minimal side effects and the highest therapeutic potential. In this framework, electrospun scaffolds incorporated with various cells or bioactive agents have provided promising vistas to repair urethral defects. Due to the biomimetic nature of these constructs, they can efficiently mimic the native cells' niches and provide essential microenvironmental cues for the safe transplantation of multiple cell types. Furthermore, these scaffolds are versatile platforms for delivering various drug molecules, growth factors, and nucleic acids. This review discusses the recent progress, applications, and challenges of electrospun scaffolds to deliver cells or bioactive agents during the urethral defect repair process. First, the current status of electrospinning in urethral tissue engineering is presented. Then, the principles of electrospinning in drug and cell delivery applications are reviewed. Finally, the recent preclinical studies are summarized and the current challenges are discussed.

Accelerating healing of excisional wound with alginate hydrogel containing naringenin in rat model.

Wounds have always been considered as one of the most common physical damages. Therefore, various researches have been conducted to find an appropriate method to improve wound healing process. Among various materials, since hydrogels have appropriate properties for wound healing, they are widely used for this purpose. In this study, to develop a potential wound dressing, different concentrations of naringenin (0%, 1%, 10% and 20%) were incorporated in alginate hydrogel followed by evaluating its characters such as morphology, swelling properties, weight loss, antibacterial activity, releasing profile of the naringenin, hemo-, and cytocompatibility. Finally, to evaluate the effect of developed hydrogels on wound healing, the full-thickness dermal wound model in rat was used. Our results provided that the prepared hydrogels have appropriate porosity (86.7 ± 5.3%) with the interconnected pores. Moreover, weight loss assessment confirmed that fabricated hydrogels have suitable biodegradability (about 89% after 14 days). MTT assay also revealed the positive effect of hydrogels on cell viabilities, and they have no toxicity effect on cells. In vivo study indicated that the prepared hydrogels had better wound closure than the gauze-treated wound (the control), and alginate/20% naringenin group had the best wound closure among other groups. All in all, this study concluded that alginate/naringenin hydrogel has positive effect on wound healing process, and it can be used to treat skin injuries in the clinic. Graphical abstract.

A tailored polylactic acid/polycaprolactone biodegradable and bioactive 3D porous scaffold containing gelatin nanofibers and Taurine for bone regeneration.

The focus of the current study was to develop a functional and bioactive scaffold through the combination of 3D polylactic acid (PLA)/polycaprolactone (PCL) with gelatin nanofibers (GNFs) and Taurine (Tau) for bone defect regeneration. GNFs were fabricated via electrospinning dispersed in PLA/PCL polymer solution, Tau with different concentrations was added, and the polymer solution converted into a 3D and porous scaffold via the thermally-induced phase separation technique. The characterization results showed that the scaffolds have interconnected pores with the porosity of up to 90%. Moreover, Tau increased the wettability and weight loss rate, while compromised the compressive strengths. The scaffolds were hemo- and cytocompatible and supported cell viability and proliferation. The in vivo studies showed that the defects treated with scaffolds filled with new bone. The computed tomography (CT) imaging and histopathological observation revealed that the PLA/PCL/Gel/Tau 10% provided the highest new bone formation, angiogenesis, and woven bone among the treatment groups. Our finding illustrated that the fabricated scaffold was able to regenerate bone within the defect and can be considered as the effective scaffold for bone tissue engineering application.

Electrospun cellulose acetate/gelatin nanofibrous wound dressing containing berberine for diabetic foot ulcer healing: in vitro and in vivo studies.

Functional wound dressing with tailored physicochemical and biological properties is vital for diabetic foot ulcer (DFU) treatment. Our main objective in the current study was to fabricate Cellulose Acetate/Gelatin (CA/Gel) electrospun mat loaded with berberine (Beri) as the DFU-specific wound dressing. The wound healing efficacy of the fabricated dressings was evaluated in streptozotocin-induced diabetic rats. The results demonstrated an average nanofiber diameter of 502 ± 150 nm, and the tensile strength, contact angle, porosity, water vapor permeability and water uptake ratio of CA/Gel nanofibers were around 2.83 ± 0.08 MPa, 58.07 ± 2.35°, 78.17 ± 1.04%, 11.23 ± 1.05 mg/cm2/hr, and 12.78 ± 0.32%, respectively, while these values for CA/Gel/Beri nanofibers were 2.69 ± 0.05 MPa, 56.93 ± 1°, 76.17 ± 0.76%, 10.17 ± 0.21 mg/cm2/hr, and 14.37 ± 0.42%, respectively. The antibacterial evaluations demonstrated that the dressings exhibited potent antibacterial activity. The collagen density of 88.8 ± 6.7% and the angiogenesis score of 19.8 ± 3.8 obtained in the animal studies indicate a proper wound healing. These findings implied that the incorporation of berberine did not compromise the physical properties of dressing, while improving the biological activities. In conclusion, our results indicated that the prepared mat is a proper wound dressing for DFU management and treatment.

A promising wound dressing based on alginate hydrogels containing vitamin D3 cross-linked by calcium carbonate/d-glucono-δ-lactone.

In the present study, we fabricated vitamin D3-loaded alginate hydrogel and assessed its wound healing capability in the animal model. The various concentrations of vitamin D3 were added to the pre-dissolved sodium alginate in deionized water and cross-linked by calcium carbonate in combination with d-glucono-δ-lactone. The microstructure, swelling behavior, weight loss, hemo- and cytocompatibility of the fabricated hydrogels were evaluated. In the last stage, the therapeutic efficacy of the prepared hydrogels was evaluated in the full-thickness dermal wound model. The scanning electron microscopy images showed that the prepared hydrogel was highly porous with the porosity of 89.2 ± 12.5% and contained the interconnected pores. Weight loss assessment showed that the prepared hydrogel is biodegradable with the weight loss percentage of about 89% in 14 days. The results showed that the prepared hydrogels were hemo- and cytocompatible. The animal study results implied that alginate hydrogel/3000 IU vitamin D3 group exhibited the highest wound closure present which was statistically significant than the control group (p < 0.05). Moreover, the histological examinations revealed that hydrogel containing 3000 IU vitamin D3 had the best performance and induced the highest re-epithelialization and granular tissue formation. All in all, this study suggests that alginate hydrogels with 3000 IU vitamin D3 can be exploited as a potential wound dressing in skin tissue engineering.

Kaolin-loaded chitosan/polyvinyl alcohol electrospun scaffold as a wound dressing material: in vitro and in vivo studies.

OBJECTIVE: To evaluate the application of a fabricated dressing containing kaolin for skin regeneration in a rat model of excisional wounds. METHOD: In the present study, kaolin was loaded into electrospun polyvinyl alcohol (PVA)/chitosan polymer blend to develop a composite nanofibrous dressing. To make the yarns, kaolin with weight ratio of 5% was added to PVA/chitosan polymer blend and subsequently formed into nanofibres using the electrospinning method. Scaffolds were evaluated for to their microstructure, mechanical properties, surface wettability, water vapour transmission rate, water-uptake capacity, blood uptake capacity, blood compatibility, microbial penetration test, the number of colonies, and cellular response with the L929 cell line. Rats with full-thickness excisional wounds were treated with kaolin-containing and kaolin-free dressings. RESULTS: The study showed that rats treated with the kaolin-incorporated mats demonstrated a significant closure to nearly 97.62±4.81% after 14 days compared with PVA/chitosan and the sterile gauze, which showed 86.15±8.11% and 78.50±4.22% of wound closure, respectively. The histopathological studies showed that in the PVA/chitosan/kaolin group, dense and regular collagen fibres were formed, while wounds treated with sterile gauze or PVA/chitosan scaffolds had random and loose collagen fibres. CONCLUSION: Our results show the potential applicability of PVA/chitosan/kaolin scaffolds as a wound care material.

Impact of exosome-loaded chitosan hydrogel in wound repair and layered dermal reconstitution in mice animal model.

Combat or burn injuries are associated with a series of risks, such as microbial infection, an elevated level of inflammatory response, and pathologic scar tissue formation, which significantly postpone wound healing and also lead to impaired repair. Skin engineering for wound healing requires a biomimetic dressing substrate with ideal hydrophilicity, holding antioxidant and antimicrobial properties. In addition, available bioactive specification is required to reduce scar formation, stimulate angiogenesis, and improve wound repair. In this study, we successfully fabricated chitosan (Ch)-based hydrogel enriched with isolated exosome (EXO) from easy-accessible stem cells, which could promote fibroblast cell migration and proliferation in vitro. Full-thickness excisional wound model was used to investigate the in vivo dermal substitution ability of the fabricated hydrogel composed Ch and EXO substrates. Our finding confirmed that the wounds covered with Ch scaffold containing isolated EXO have nearly 83.6% wound closure ability with a high degree of re-epithelialization, whereas sterile gauze showed 51.5% of reduction in wound size. In summary, obtained results imply that Ch-glycerol-EXO hydrogel construct can be utilized at the full-thickness skin wound substitution and skin tissue engineering.

Exosome loaded alginate hydrogel promotes tissue regeneration in full-thickness skin wounds: An in vivo study.

Wound healing is known as one of the most complicated biological processes for injured skin caused by surgical, trauma, burns, or diabetic diseases, which causes a nonfunctioning mass of fibrotic tissue. Recent reports have suggested that exosomes (EXOs) secreted by this type of stem cells may contribute to their paracrine effect. In this study, the EXOs were isolated from the supernatant of cultured adipose-derived stem cells (ADSCs) via ultracentrifugation and filtration. The EXO loaded in the alginate-based hydrogel was used as a bioactive scaffold to preserve the EXO in the wound site in the animal model. The physical and biochemical properties of EXO loaded Alg hydrogel were characterized and results proved that fabricated structure was biodegradable and biocompatible. This bioactive wound dressing technique has significantly improved wound closure, collagen synthesis, and vessel formation in the wound area. Results offer a new viewpoint and a cell-free therapeutic strategy, for wound healing through the application of the composite structure of EXO encapsulated in alginate hydrogel.

A novel polycaprolactone/carbon nanofiber composite as a conductive neural guidance channel: an in vitro and in vivo study.

The current study aimed to investigate the potential of carbon nanofibers to promote peripheral nerve regeneration. The carbon nanofiber-imbedded scaffolds were produced from polycaprolactone and carbon nanofibers using thermally induced phase separation method. Electrospinning technique was utilized to fabricate polycaprolactone/collagen nanofibrous sheets. The incorporation of carbon nanofibers into polycaprolactone's matrix significantly reduced its electrical resistance from 4.3 × 109 ± 0.34 × 109 Ω to 8.7 × 104 ± 1.2 × 104 Ω. Further in vitro studies showed that polycaprolactone/carbon nanofiber scaffolds had the porosity of 82.9 ± 3.7% and degradation rate of 1.84 ± 0.37% after 30 days and 3.58 ± 0.39% after 60 days. The fabricated scaffolds were favorable for PC-12 cells attachment and proliferation. Neural guidance channels were produced from the polycaprolactone/carbon nanofiber composites using water jet cutter machine then incorporated with PCL/collagen nanofibrous sheets. The composites were implanted into severed rat sciatic nerve. After 12 weeks, the results of histopathological examinations and functional analysis proved that conductive conduit out-performed the non-conductive type and induced no toxicity or immunogenic reactions, suggesting its potential applicability to treat peripheral nerve damage in the clinic.

Sciatic nerve regeneration by using collagen type I hydrogel containing naringin.

In the present study, collagen hydrogel containing naringin was fabricated, characterized and used as the scaffold for peripheral nerve damage treatment. The collagen was dissolved in acetic acid, naringin added to the collagen solution, and cross-linked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide powder (EDC; 0.10 mM) to form the hydrogel. The microstructure, swelling behavior, biodegradation, and cyto/hemocompatibility of the fabricated hydrogels were assessed. Finally, the healing efficacy of the prepared collagen hydrogel loaded with naringin on the sciatic nerve crush injury was assessed in the animal model. The characterization results showed that the fabricated hydrogels have a porous structure containing interconnected pores with the average pore size of 90 µm. The degradation results demonstrated that about 70% of the primary weight of the naringin loaded hydrogel had been lost after 4 weeks of storage in PBS. The in vitro study showed that the proliferation of Schwann cells on the collagen/naringin hydrogel was higher than the control group (tissue culture plate) at both 48 and 72 h after cell seeding and even significantly higher than pure collagen 72 h after cell seeding (*p < 0.005, **p < 0.001). The animal study implied that the sciatic functional index reached to -22.13 ± 3.00 at the end of 60th days post-implantation which was statistically significant (p < 0.05) compared with the negative control (injury without the treatment) (-82.60 ± 1.06), and the pure collagen hydrogel (-59.80 ± 3.20) groups. The hot plate latency test, the compound muscle action potential, and wet weight-loss of the gastrocnemius muscle evaluation confirmed the positive effect of the prepared hydrogels on the healing process of the induced nerve injury. In the final, the histopathologic examinations depicted that the collagen/naringin hydrogel group reduced all the histological changes induced from the nerve injury and showed more resemblance to the normal sciatic nerve, with well-arranged fibers and intact myelin sheath. The overall results implied that the prepared collagen/naringin hydrogel can be utilized as a sophisticated alternative to healing peripheral nerve damages.

Fabrication of Poly(L-Lactic Acid)/Chitosan Scaffolds by Solid-Liquid Phase Separation Method for Nerve Tissue Engineering: An In Vitro Study on Human Neuroblasts.

Polymeric scaffolds that support neural cell behaviors are attracting more attention. In the present study, solid-liquid phase separation technique is used to fabricate scaffolds made of poly(L-lactic acid) (PLLA) and chitosan (CS) blends to mimic both cellular microenvironment and anatomical structure of nerve tissue. The fabricated scaffolds favor characteristics of both natural and synthetic polymers. Different tests and assays including physical and mechanical ones (in vitro degradation rate, free radical release, hydrophilicity, and porosity measurements, microstructure observation, and mechanical tests) and cellular assays (cell attachment measurement and viability assessment) suggest that blend scaffolds prepared with this method support nerve cells for tissue engineering applications adequately and even better than scaffolds prepared with the same method but from pure PLLA or CS.

Extracellular micro/nanovesicles rescue kidney from ischemia-reperfusion injury.

Acute renal failure (ARF) is a clinical challenge that is highly resistant to treatment, and its high rate of mortality is alarming. Ischemia-reperfusion injury (IRI) is the most common cause of ARF. Especially IRI is implicated in kidney transplantation and can determine graft survival. Although the exact pathophysiology of renal IRI is unknown, the role of inflammatory responses has been elucidated. Because mesenchymal stromal cells (MSCs) have strong immunomodulatory properties, they are under extensive investigation as a therapeutic modality for renal IRI. Extracellular vesicles (EVs) play an integral role in cell-to-cell communication. Because the regenerative potential of the MSCs can be recapitulated by their EVs, the therapeutic appeal of MSC-derived EVs has dramatically increased in the past decade. Higher safety profile and ease of preservation without losing function are other advantages of EVs compared with their producing cells. In the current review, the preliminary results and potential of MSC-derived EVs to alleviate kidney IRI are summarized. We might be heading toward a cell-free approach to treat renal IRI.

Unrestricted Somatic Stem Cells Loaded in Nanofibrous Conduit as Potential Candidate for Sciatic Nerve Regeneration.

Motor and sensory recovery following critical size peripheral nerve defects is often incomplete. Although nerve grafting has been proposed as the gold standard, it is associated with several disadvantages. Here we report a novel approach to peripheral nerve repair using Human Unrestricted Somatic Stem Cells (USSC) delivered through an electrospun neural guidance conduit. Conduits were produced from PCL and gelatin blend. Several in vitro methods were utilized to investigate the conduit's physicochemical and biological characteristics. Nerve regeneration was studied across a 10-mm sciatic nerve gap in Wistar rats. For functional analysis, the conduits were seeded with 3 × 104 USSCs and implanted into a 10-mm sciatic nerve defect. After 14 weeks, the results of functional recovery analysis and histopathological examinations showed that animals implanted with USSC containing conduits exhibited improved functional and histopathological recovery which was more close to the autograft group compared to other groups. Our results support the potential applicability of USSCs to treat peripheral nerve injury in the clinic.

The cardiac niche role in cardiomyocyte differentiation of rat bone marrow-derived stromal cells: comparison between static and microfluidic cell culture methods.

Due to the restricted potential of the heart to regenerate its damaged region, stem cell therapy is a promising treatment modality for myocardial infarction. It has been shown that incubation of bone marrow-derived stromal cells (BMSCs) with initial steps of cardiac differentiation in vitro, can have a significant effect on their therapeutic potential to treat myocardial infarction. Based on these well-established principals we were encouraged to study the direct co-culture of rat BMSCs with neonatal mouse almost pure cardiomyocytes (APCs) and cardiac niche cells (CNCs) in static 2D and microfluidic cell culture systems. Our results showed that the difference regarding the beating rate in isolated APCs and CNCs in both 2D and the microfluidic system was not statistically significant for 30 days. No beat rate could be observed in induced BMSCs in all groups during experiment time. Except for BMSCs cultured alone in both experimental culture conditions, data obtained from Real-time PCR analysis showed that differentiated BMSCs in all co-cultured groups expressed GATA4, Nkx2.5, CX43, cTnI, cTnT, and ß-MHC during 4 weeks. BMSCs demonstrated a higher expression of these cardiac factors in microfluidic chips than those co-cultured in 24 well plates. Moreover, immunocytochemistry (ICC), also revealed the GATA4 expression in differentiated BMSCs in all co-cultured groups. It was found that, when combined with shear stress, co-culture with cardiomyocyte can differentiate BMSCs significantly toward cardiomyocyte rather than co-culture alone.

In vitro and in vivo study of PCL/COLL wound dressing loaded with insulin-chitosan nanoparticles on cutaneous wound healing in rats model.

In the current study, insulin delivering chitosan nanoparticles were coated onto the electrospun poly (ε-caprolactone) (PCL)/Collagen (COLL) to produce a potential wound care material. Electrospun matrices were fabricated from PCL/COLL (1:1 (w/w)) solution. The insulin-loaded chitosan nanoparticles were produced by ionic gelation process and then attached onto the yarns. The dressings were investigated regarding their surface wettability, microstructure, the capacity to absorb water, water vapour permeability, mechanical properties, blood compatibility, microbial penetration, and cellular behavior. Full-thickness excisional wound model was used to assess the in vivo healing capacity of the dressings. Our data showed that after 14 days the wounds covered with PCL/COLL/Cs-Ins wound dressing could reach to nearly full wound closure compared with the sterile gauze which exhibited nearly 45% of wound size reduction. Our results suggest that fabricated scaffolds can be potentially applied in clinical practice for wound treatment.

In vitro and in vivo evaluation of electrospun cellulose acetate/gelatin/hydroxyapatite nanocomposite mats for wound dressing applications.

The present study aimed to evaluate the efficacy of cellulose acetate/gelatin/nanohydroxyapatite (CA/Gel/nHA) nanocomposite mats as the wound dressing. The dressings were prepared with electrospinning of CA/Gel solutions containing 12.5, 25 and 50 mg nHA. The dressings were evaluated regarding their water uptake capacity, morphology, tensile strength, water vapour transmission rate, wettability and cellular response with L929 cell line. The results showed that the concentration of nHA had a direct correlation with porosity, water contact angle, water uptake, water vapor transmission rate and proliferation. In vivo studies showed that all dressings had higher wound closure percent than the sterile gauze, as the control. The highest wound closure value was achieved in the CA/Gel +25 mg nHA group, which showed 93.5 ± 1.6%. The histological and the histomorphometric examinations of the wounds revealed that the CA/Gel +25 mg nHA dressing had the greatest collagen synthesis, re-epithelialization, neovascularization and also the best cosmetic appearance. Based on our finding, it could be concluded the applicability of electrospun nanofibrous CA/Gel/nHA dressings for successful wound treatment.

Regeneration of sciatic nerve crush injury by a hydroxyapatite nanoparticle-containing collagen type I hydrogel.

The current study aimed to enhance the efficacy of peripheral nerve regeneration using a hydroxyapatite nanoparticle-containing collagen type I hydrogel. A solution of type I collagen, extracted from the rat tails, was incorporated with hydroxyapatite nanoparticles (with the average diameter of ~212 nm) and crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) to prepare the hydrogel. The Schwann cell cultivation on the prepared hydrogel demonstrated a significantly higher cell proliferation than the tissue culture plate, as positive control, after 48 h (n = 3, P < 0.005) and 72 h (n = 3, P < 0.01). For in vivo evaluation, the prepared hydrogel was administrated on the sciatic nerve crush injury in Wistar rats. Four groups were studied: negative control (with injury but without interventions), positive control (without injury), collagen hydrogel and hydroxyapatite nanoparticle-containing collagen hydrogel. After 12 weeks, the administration of hydroxyapatite nanoparticle-containing collagen significantly (n = 4, P < 0.005) enhanced the functional behavior of the rats compared with the collagen hydrogel and negative control groups as evidenced by the sciatic functional index, hot plate latency and compound muscle action potential amplitude measurements. The overall results demonstrated the applicability of the produced hydrogel for the regeneration of peripheral nerve injuries.