Journal of Tissue Engineering and Regenerative Medicine最新文献

Tissue-engineered blood vessels (TEBVs) show significant therapeutic potential for replacing diseased blood vessels. Vascular smooth muscle cells (VSMCs) derived from human induced pluripotent stem cells (hiPSCs) via embryoid body (EB)-based differentiation, are promising seed cells to construct TEBVs. However, obtaining sufficient high-quality hiPSC-VSMCs remains challenging. Stem cells are located in a niche characterized by hypoxia. Hence, we explored molecular and cellular functions at different induction stages from the EB formation commencement to the end of directed differentiation under normoxic and hypoxic conditions, respectively. Hypoxia enhanced the formation, adhesion and amplification rates of EBs. During directed differentiation, hiPSC-VSMCs exhibited increased cell viability under hypoxic conditions. Moreover, seeding hypoxia-pretreated cells on biodegradable scaffolds, facilitated collagen I and elastin secretion, which has significant application value for TEBV development. Hence, we proposed that hypoxic treatment during differentiation effectively induces proliferative hiPSC-VSMCs, expanding high-quality seed cell sources for TEBV construction.

Utilizing recent advances in human induced pluripotent stem cell (hiPSC) technology, nonlinear analysis and machine learning we can create novel tools to evaluate drug-induced cardiotoxicity on human cardiomyocytes. With cardiovascular disease remaining the leading cause of death globally it has become imperative to create effective and modern tools to test the efficacy and toxicity of drugs to combat heart disease. The calcium transient signals recorded from hiPSC-derived cardiomyocytes (hiPSC-CMs) are highly complex and dynamic with great degrees of response characteristics to various drug treatments. However, traditional linear methods often fail to capture the subtle variation in these signals generated by hiPSC-CMs. In this work, we integrated nonlinear analysis, dimensionality reduction techniques and machine learning algorithms for better classifying the contractile signals from hiPSC-CMs in response to different drug exposure. By utilizing extracted parameters from a commercially available high-throughput testing platform, we were able to distinguish the groups with drug treatment from baseline controls, determine the drug exposure relative to IC50 values, and classify the drugs by its unique cardiac responses. By incorporating nonlinear parameters computed by phase space reconstruction, we were able to improve our machine learning algorithm's ability to predict cardiotoxic levels and drug classifications. We also visualized the effects of drug treatment and dosages with dimensionality reduction techniques, t-distributed stochastic neighbor embedding (t-SNE). We have shown that integration of nonlinear analysis and artificial intelligence has proven to be a powerful tool for analyzing cardiotoxicity and classifying toxic compounds through their mechanistic action.

Cornea as the outermost layer of the eye is at risk of various genetic and environmental diseases that can damage the cornea and impair vision. Corneal transplantation is among the most applicable surgical procedures for repairing the defected tissue. However, the scarcity of healthy tissue donations as well as transplantation failure has remained as the biggest challenges in confront of corneal grafting. Therefore, alternative approaches based on stem-cell transplantation and classic regenerative medicine have been developed for corneal regeneration. In this review, the application and limitation of the recently-used advanced approaches for regeneration of cornea are discussed. Additionally, other emerging powerful techniques such as 5D printing as a new branch of scaffold-based technologies for construction of tissues other than the cornea are highlighted and suggested as alternatives for corneal reconstruction. The introduced novel techniques may have great potential for clinical applications in corneal repair including disease modeling, 3D pattern scheming, and personalized medicine.

Large animal testing and clinical trials using bioengineered bladder for augmentation have revealed that large grafts fail due to insufficient blood supply. To address this critical issue, an in vivo staged implant strategy was developed and evaluated to create autologous, vascularized bioengineered bladder tissue with potential for clinical translation. Pig bladders were used to create acellular urinary bladder matrices (UBMs), which were implanted on the rectus abdominus muscles of rats and pigs to generate cellular and vascular grafts. Rectus-regenerated bladder grafts (rrBGs) were highly cellularized and contained an abundance of CD31-positive blood vessels, which were shown to be functional by perfusion studies. Muscle patterns within grafts showed increased smooth muscle formation over time and specifically within the detrusor compartment, with no evidence of striated muscle. Large, autologous rrBGs were transplanted to the pig bladder after partial cystectomy and compared to transplantation of control UBMs at 2 weeks and 3 months post-transplant. Functional, ink-perfused blood vessels were found in the central portion of all rrBGs at 2 weeks, while UBM grafts were significantly deteriorated, contracted and lacked central cellularization and vascularization. By 3 months, rrBGs had mature smooth muscle bundles and were morphologically similar to native bladder. This staged implantation technique allows for regeneration and harvest of large bladder grafts that are morphologically similar to native tissue with functional vessels capable of inosculating with host bladder vessels to provide quick perfusion to the central area of the large graft, thereby preventing early ischemia and contraction.

Muscular dysplasia is the key factor in influencing surgical outcomes in patients with cleft lip/palate. In this research, we attempted to evaluate a new acellular dermal matrix (ADM) as a substitute for reconstruction of the orbicularis oris muscle with growth factors such as Insulin-Like Growth Factor I (IGF-I), vascular endothelial growth factor (VEGF) in a rabbit model. 30 male New Zealand Rabbits (2–3 m, 1700–2000 g) were divided into four groups as follows; a group in which the orbicularis oris muscle of the upper lip was implanted with ADM, a group in which the orbicularis oris muscle of the upper lip was implanted with ADM + IGF-I + VEGF, a group in which the upper lip was operated without implantation of an ADM scaffold, and a normal upper lip for comparison. Macroscopic observation, histological evaluation, and immunohistochemistry were employed to evaluate levels of the muscle regeneration, vascularization, and inflammation at 1, 2, 4, 6, and 12 weeks after the operation. All wounds healed well without infection, immune rejection and so on. Histological evaluation showed that ADM was totally degraded and replaced by connective tissue. The area in which the ADM scaffold was coated with growth factors show a significant increase in the formation of new myofibers after injury, and the vascularization improved compared to the control group and the normal group. In regard to the degrees of inflammation, there were no notable differences among the groups. In conclusion, Our study indicated that ADM grafts combined with IGF-I and VEGF have potential advantages in alleviating muscular dysplasia in cleft lip treatment.

Recently, tissue engineering and regenerative medicine have received significant attention with outstanding advances. The main scope of this technology is to recover the damaged tissues and organs or to maintain and improve their function. One of the essential fields in tissue engineering is scaffold designing and construction, playing an integral role in damaged tissues reconstruction and repair. However, premature ovarian failure (POF) is a disorder causing many medical and psychological problems in women. POF treatment using tissue engineering and various scaffold has recently made tremendous and promising progress. Due to the importance of the subject, we have summarized the recently examined scaffolds in the treatment of POF in this review.

The moderate static magnetic fields (SMFs) have been used here as a non-invasive tool to study their manipulative effects on the olfactory ensheathing cells (OECs) activity, growth, morphology, and migration in culture. The OECs are involved in the regeneration of primary olfactory sensory neurons and migration into the central nervous system to repair axons damaged by infection, injury, etc., that play a pivotal role in complementary regenerative medicine. Here, OECs were isolated from the olfactory bulb and cultured to confluence. An in vitro wound healing model was formed and exposed to either parallel (PaSMF) or perpendicular (PeSMF) SMF at intensities of 30, 50, and 70 mT, and cells' morphology, podia formation, proliferation, and migration were studied by time-lapse recording. The SMFs were not cytotoxic at the intensity and exposure time applied here. The exposure of cells to 70 mT PaSMF and PeSMF increased the formation of lamellipodia and filopodia, cell migration speed, and direction of the scratch forefront cells, significantly. Treatment of cells with 70 mT PaSMF and PeSMF increased cell divisions, while 30 mT PaSMF decreased it. SMF effects on OECs division, motility, migratory direction, and velocity indicate its effect on various aspects of cell physiology and signaling at atomic and molecular levels, and have a role in tissue regeneration that involves microtubules and actin filaments formation and rearrangements. Thus, the exposure of OECs with moderate SMF might be considered a promising noninvasive approach to remotely manipulate normal and stem cell activities for therapeutic regenerative purposes in various tissues including the central nervous system.

Despite several advances in chronic wound management, natural product based scaffolds with high exude absorption and mechanical strength are still a hotspot in the medical field. Thus, present study illustrates the fabrication of agarose (AG; 10% w/v)/polyvinyl alcohol 12% w/v) based multifunctional nanofibrous electrospun scaffolds. Zinc citrate (1%, 3% and 5% w/w of the polymer) was used as a potential antibacterial agent. The fabricated scaffolds exhibit a swelling of ∼550% in phosphate buffer saline and mechanical strength of 10.11 ± 0.31 MPa which is suitable for most of the wound healing applications that require high strength. In vitro study revealed an increased migration and proliferation of L929 fibroblasts with AG blends when compared to the control. The fabricated scaffolds exhibited antibacterial properties against both Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative) bacterial strains. Hence, a multifunctional (ability to protect wounds from bacterial infections along with effective swelling and mechanical support), natural product based, eco-friendly scaffold to serve as a potential wound dressing material has been successfully fabricated.

This study evaluated the effect of low (20 and 40 mV/mm) intensities of electrical stimulation on the proliferation and migration of skin fibroblasts from diabetic donors. We also examined the effect of electrical stimulation on modulating the capacity of fibroblasts to contract collagen gel, express alpha-smooth muscle actin, and secrete proteolytic enzymes involved in regulating extracellular matrix synthesis and degradation. Our study shows that 20 and 40 mV/mm of stimulation increased the growth of fibroblasts extracted from diabetic patients but not from non-diabetic donors. Electrical stimulation increased the migration of diabetic fibroblasts, their capacity to contract collagen gel, and the expression of alpha-smooth muscle actin and promoted different proteolytic enzymes involved in accelerating wound healing. Overall results confirm the effectiveness of electrical stimulation in modulating the wound healing activities of fibroblasts extracted from diabetic skin donors. This study, therefore, suggests the possible use of electrical stimulation to promote diabetic foot ulcer healing by stimulating the wound healing properties of skin fibroblasts.

MicroRNA-21 (miR-21) can induce proliferation and differentiation of mesenchymal stem cells (MSCs) to promote bone formation, we therefore aimed to investigate whether exosomes derived from miR-21 overexpressing adipose tissue-derived MSCs (AD-MSCs) could improve spine osteoporosis in ankylosing spondylitis (AS) mice. Cultured AD-MSCs were transfected with lentivirus vectors containing miR-21 or control vector, and the supernatant was centrifugated and filtrated to harvest the exosomes (miR-21-Exos or vector-Exos). BALB/c mice were immunized with cartilage proteoglycan to establish proteoglycan-induced ankylosing spondylitis (PGIA) model. Six weeks later, PGIA mice were further injected with miR-21-Exos or vector-Exos. Transfection of miR-21 in AD-MSCs significantly enhanced miR-21 levels in AD-MSCs and their exosomes. miR-21-Exos showed concentration-dependent protective effect against spine osteoporosis in PGIA mice, evidenced by increased bone mineral content and bone mineral density, reduced number of osteoclasts, decreased content of deoxypyridinoline in the urine, decreased content of tartrate-resistant acid phosphatase (TRACP)-5b and cathepsin K in the serum, and down-regulated interleukin (IL)-6 expression in the spine, whereas vector-Exos did not show any treatment benefit. The above findings indicate that miR-21-Exos could be utilized to treat spine osteoporosis in AS.