Comparison of effects of P-coumaric acid and coumarin on colorectal cancer cell line by inducing apoptosis and autophagy.

Objective: Autophagy, as a cellular pathway involved in removing damaged proteins and organelles, performs a vital function in the homeostasis and fate of cells. Natural compounds of coumarin (CO) are found in a variety of herbs. Due to their many medicinal properties, including antitumor and anti-proliferative activity, they are involved in apoptosis and autophagy processes. This investigation desired to analyze the apoptotic and autophagic effects of p-coumaric acid (PCA) and CO on HT-29 cells cultured in fibrin hydrogel. Materials and Methods: Cell viability and apoptotic and autophagic changes were evaluated by MTT assay, Acridine Orange, 4',6-diamidino-2-phenylindole (DAPI), and monodansylcadaverine (MDC) staining. The expression Bax, Bad, Bcl2, Lc3, Beclin-1, P53 and Atg5 was respectively measured by qRT-PCR and Western blotting. Results: CO (IC50=25 µM) and PCA (IC50=150 µM) had a dose- and time-dependent cytotoxic effect in HT-29 cells. So, the cytotoxic effects of CO were significantly higher than PCA and these differences were also evident in cell morphology investigations. The data illustrated a high expression of pro-apoptotic and pro-autophagic genes and a declined expression of anti-apoptotic and anti-autophagic genes. Conclusion: CO (that was more potent) and p-coumaric acid-induced autophagy via PI3K/Akt/mTOR and AMPK/mTOR signaling on HT-29 cells.

Ex vivo expansion of hematopoietic stem cells in two/ three-dimensional co-cultures with various source of stromal cells.

The ex vivo expansion of hematopoietic stem cells, with both high quantities and quality, is considered a paramount issue in cell and gene therapy for hematological diseases. Complex interactions between the bone marrow microenvironment and hematopoietic stem cells reveal the importance of using 2D and 3D coculture as a physiological system simulator in the proliferation, differentiation, and homeostasis of HSCs. Herein, the capacity of mesenchymal stem cells derived from different sources to support the expansion and maintenance of HSPC was compared with each other. We evaluated the fold increase of HSPC, CD34 marker expression, cytokine secretion profile of different MSCs, and the frequency of hematopoietic colony-forming unit parameters. Our results show that there was no significant difference between adipose tissue-MSC, Wharton jelly-MSC, and Endometrial-MSCs in HSPC expansion (fold increase: 34.74±4.38 in Wj-MSC, 32.22±5.07 in AD-MSC, 25.9±1.27 in En-MSCs); However, there were significantly more than the expansion media alone (4.4±0.69). The results obtained from the cytokine secretion analysis also confirm these results. Also, there were significant differences in the clonogenicity of Wj-MSC, En-MSCs, and expansion media (CFU-GEMM: 7±1.73, 2.3±1.15, and 2.3±1.52), which indicated that Wj-MSC could significantly maintain the primitive state. As a result, using Wj-mesenchymal stem cells on a 3D coculture system effectively increases the HSPC expansion and maintains the colonization potential of hematopoietic stem cells.

Evaluation of lncRNA Expression During the Differentiation of Mesenchymal Stem Cells to Insulin-Secreting Progenitors.

Diabetes mellitus is a metabolic disease caused by a defect in insulin secretion, insulin function, or both that destroys pancreatic islet beta cells. There is ample evidence that long non-coding RNAs (lncRNAs) play a vital role in cell formation and differentiation. The present study aims to investigate the expression pattern of specific lncRNAs in mesenchymal stem cell (MSC) differentiation into insulin-producing beta cell (IPCs) progenitors for cell therapy purposes. MSCs were extracted from human umbilical cord Wharton jelly (hWJ-MSCs) using the explant method and cultured in two-dimensional (2D) and three-dimensional (3D) media on polylactic acid/Wax (PLA/Wax) nanofibrous scaffold using a three-step protocol containing CHIR99021 small molecules and Indolactam V. At the end of each differentiation step, immunocytochemistry and qRT-PCR were used to confirm the differentiation at the protein and RNA levels and the expression changes of six selective lncRNAs were evaluated by qRT-PCR. The results indicated that the expression of the selected lncRNAs was significantly altered during the differentiation process into beta progenitor cells, indicating their potential role in regulating the IPC differentiation process. More specifically, all of the desired lncRNAs demonstrated a significant increase during the beta cell differentiation, with HI-LNC71 and HI-LNA12 experiencing the highest expression in the produced Beta cell progenitors respectively (p<0.0001). These results can be valuable in tissue engineering and treatment studies by replacing beta precursor cells to control diabetic patients.

Enhancement of nerve regeneration through schwann cell-mediated healing in a 3D printed polyacrylonitrile conduit incorporating hydrogel and graphene quantum dots: a study on rat sciatic nerve injury model.

Despite recent technological advancements, effective healing from sciatic nerve damage remains inadequate. Cell-based therapies offer a promising alternative to autograft restoration for peripheral nerve injuries, and 3D printing techniques can be used to manufacture conduits with controlled diameter and size. In this study, we investigated the potential of Wharton's jelly-derived mesenchymal stem cells (WJMSCs) differentiated into schwann cells, using a polyacrylonitrile (PAN) conduit filled with fibrin hydrogel and graphene quantum dots (GQDs) to promote nerve regeneration in a rat sciatic nerve injury model. We investigated the potential of WJMSCs, extracted from the umbilical cord, to differentiate into schwann cells and promote nerve regeneration in a rat sciatic nerve injury model. WJMSCs were 3D cultured and differentiated into schwann cells within fibrin gel for two weeks. A 3 mm defect was created in the sciatic nerve of the rat model, which was then regenerated using a conduit/fibrin, conduit covered with schwann cells in fibrin/GQDs, GQDs in fibrin, and a control group without any treatment (n= 6/group). At 10 weeks after transplantation, motor and sensory functions and histological improvement were assessed. The WJMSCs were extracted, identified, and differentiated. The differentiated cells expressed typical schwann cell markers, S100 and P75.In vivoinvestigations established the durability and efficacy of the conduit to resist the pressures over two months of implantation. Histological measurements showed conduit efficiency, schwann cell infiltration, and association within the fibrin gel and lumen. Rats treated with the composite hydrogel-filled PAN conduit with GQDs showed significantly higher sensorial recovery than the other groups. Histological results showed that this group had significantly more axon numbers and remyelination than others. Our findings suggest that the conduit/schwann approach has the potential to improve nerve regeneration in peripheral nerve injuries, with future therapeutic implications.

Simultaneous Administration of Berberine and Transplantation of Endometrial Stem Cell-Derived Insulin Precursor Cells on a Nanofibrous Scaffold to Treat Diabetes Mellitus in Mice.

Today, significant success has been achieved in treating diabetes with cell therapy derived from various sources of stem and progenitors. The replacement of beta cells is one of the new diabetes treatment methods. To this end, the production of pancreatic beta precursors in cell culture has created an important research field for diabetes treatment. Endometrial stem cells were isolated using an enzymatic method, and after their identity was confirmed using a flow cytometry and differentiation potential assay, the isolated cells were cultured on an electrospun PCL/CS scaffold. Endometrial cells were differentiated into insulin-producing cells (IPCs), and gene expression was analyzed using the qRT-PCR and immunofluorescence to confirm the creation of IPCs. Then, IPCs on the scaffold along with berberine were applied to 5 groups of diabetic mice, and after 6 weeks, insulin, blood glucose, and weight of the animals were measured. The findings revealed that pancreatic markers were significantly expressed in IPCs compared to control cells. In addition, when compared to the control group and scaffolds, the receiving group of IPCs on scaffolds had a significant improvement (p ≤ 0.0015), and this improvement increased with the addition of berberine (decrease in blood sugar (133 mg/dL), and an increase in weight (5/39 g) and insulin (2.29 MIU/L). Thus, tissue engineering is a promising new strategy for treating diabetes and can be used in the future for cell therapy and suitable drugs for diabetic patients.

Cancer cells same as zombies reprogram normal cells via the secreted microenvironment.

The cancer microenvironment plays a crucial role in promoting metastasis and malignancy even in normal cells. In the present study, the effect of acidic and conditioned media of cancer cells (MDA-MB-231), separately and in combination, was studied for the first time on the cell death mechanisms and DNA methylation of normal fibroblasts (NIH/3T3). Cell survival of conditioned media was rescued by the addition of acidic media to conditioned media, as shown by the results. Cell metabolic activity is deviated in a direction other than the Krebs cycle by acidic media The mitochondrial metabolic activity of all groups was enhanced over time, except for acidic media. Unlike the highest amount of ROS in conditioned media, its level decreased to the level of acidic media in the combination group. Furthermore, cells were deviated towards autophagy, rather than apoptosis, by the addition of acidic media to the conditioned media, unlike the conditioned media. Global DNA methylation analysis revealed significantly higher DNA hypomethylation in acidic media than in normal and combination media. Not only were cells treated with conditioned media rescued by acidic media, but also DNA hypomethylation and apoptosis in the combination group were decreased through epigenetic modifications. The acidic and conditioned media produced by cancer cells can remotely activate malignant signaling pathways, much like zombies, which can cause metabolic and epigenetic changes in normal cells.

Toxicity and autophagy effects of fluorinated cycloplatinated(II) complex bearing dppm ligand on cancer cells in in-vitro culture and in-silico prediction.

Toxicity and autophagy effects of a new complex of platinum II (CPC) were evaluated on HeLa cells cultured on a PCL/gelatin electrospinning scaffold. HeLa cells were treated with CPC on the first, third, and fifth days and the concentration of IC50 was determined. The autophagic and apoptotic effects of CPC were examined by MTT assay, Acridine Orange, Giemsa, DAPI, MDC, real-time PCR, Western blot testing, and molecular docking. The cell viability was obtained on days 1, 3, and 5 as much as 50, 7.28, and 19%, respectively with a concentration of IC50 (100µM) of CPC. The staining results indicated that the treatment of HeLa cells with CPC had antitumor and autophagic effects. Results of RT-PCR showed that the expression of BAX, BAD, P53, and LC3 genes was significantly increased in the sample treated with IC50 concentration compared to the control sample whereas the expression of BCL2, mTOR, and ACT genes in cells was significantly decreased compared to the control group. Also, these results were confirmed by Western blotting. The data indicated the induction of apoptotic death and autophagy in the studied cells. The new compound of CPC has antitumor effects.

Using Odontoblasts Derived from Dog Endometrial Stem Cells Encapsulated in Fibrin Gel Associated with BMP-2 in a Rat Pulp-Capping Model.

This study aimed to treat dental injuries by utilizing one of the most advanced tissue engineering techniques. In this study, an in vitro model was employed to investigate the proliferation and odontogenic differentiation of canine endometrial stem cells (C-EnSCs). Furthermore, the dentin regeneration potential of odontoblast like-cells (OD) derived from C-EnSCs was assessed in rats. The C-EnSCs were isolated by the enzymatic method and identified by flow cytometry. The C-EnSCs were encapsulated in fibrin gel associated with signaling factors to create the proper conditions for cell growth and differentiation. Then, the OD cells were associated with bone morphologic protein-2 (BMP-2) to promote dentin formation in vivo. The animal model used to evaluate the regenerative effect of cells and biomaterials included the preparation of the left maxillary first molar of rats for direct pulp capping operation. Animals were divided into four groups: group 1, a control group without any treatment, group 2, which received fibrin, group 3, which received fibrin with ODs (fibrin/ODs), and group 4, which received fibrin with ODs and BMP-2 (fibrin/ODs/BMP-2). The morphological observations showed the differentiation of C-EnSCs into adipose, bone, neural cells, and ODs. Furthermore, the histomorphometric data of the treated teeth showed how fibrin gel and BMP2 at a concentration of 100 ng/mL provided an optimal microenvironment for regenerating dentin tissue in rats, which was increased significantly with the presence of OD cells within eight weeks. Our study showed that using OD cells derived from C-EnSCs encapsulated in fibrin gel associated with BMP2 can potentially be an appropriate candidate for direct pulp-capping and dentin regeneration.

Encapsulation of human endometrial stem cells in chitosan hydrogel containing titanium oxide nanoparticles for dental pulp repair and tissue regeneration in male Wistar rats.

This study aimed to determine the impact of human endometrial stem cells (EnSCs) and titanium oxide nanoparticles (TiO2 NPs) on dental pulp repair and regeneration in an animal model through dentine development and tissue regeneration. The EnSCs were put on a three-dimensional (3D) chitosan scaffold containing TiO2 NPs after obtaining and purifying the collagenase enzyme. Pulps were exposed on the maxillary left first molar of all rats followed by direct pulp capping with the experimental scaffolds, as follows. Groups were: 1, control group without any treatment; 2, chitosan group (CS); 3, chitosan group with stem cells (CS/SCs); 4, chitosan group with stem cells and TiO2 NPs (CS/EnSCs/TiO2). Glass ionomer was used as a sealant in all groups. The teeth were extracted and histologically evaluated after 8 weeks. The quality and amount of dentine in the CS/EnSCs/TiO2 group were higher than in the other groups. The combination of EnSCs with TiO2 NPs and 3D chitosan scaffolds had a synergistic effect on each other, evidencing increased speed and quality of dentine formation. Using EnSCs with TiO2 NPs on a 3D chitosan scaffold can be a suitable combination for direct pulp capping and dentine regeneration in a rat molar tooth model.

Preparation, Characterization, and Wound Healing Assessment of Curcumin-Loaded M-MOF (M = Cu, Zn)@Polycaprolactone Nanocomposite Sponges.

The fabrication of multifunctional scaffolds has attracted much attention in biological fields. In this research, some novel composites of Cu(II) or Zn(II) metal-organic framework (M-MOF) and polycaprolactone (PCL), M-MOF@PCL, have been fabricated as multifunctional scaffolds for application in the tissue engineering (TE) field. The porous three-dimensional sponges were prepared by the salt leaching method. Then, the M-MOF@PCL composite sponges have been prepared by in situ synthesis of M-MOF in the presence of the as-obtained PCL sponge to gain a new compound with proper features for biological applications. Finally, curcumin was attached to the M-MOF@PCL as a bioactive compound that can act as a wound-healing agent, anti-oxidant, and anti-inflammatory. The presence of the M-MOF in final composites was investigated by different methods such as FTIR (Fourier-transform infrared), XRD (X-ray diffraction), SEM (scanning electron microscope), EDS (energy-dispersive X-ray spectroscopy), and TEM (transmission electron microscope). SEM images confirmed the porous structure of the as-obtained composites. According to the EDS and TEM images, M-MOFs were uniformly incorporated throughout the PCL sponges. The water sorption capacities of the blank PCL, Cu-MOF@PCL, and Zn-MOF@PCL were determined as 56%, 155%, and 119%, respectively. In vivo investigation on a third-degree burn model in adult male Wistar rats exhibited an accelerated wound healing for Cu-MOF@PCL compared to with Zn-MOF@PCL and the control group.

New Hg(II) coordination polymers based on a thioimidazole ligand with good performance to detoxify Hg(II) and reversibly capture iodine.

In the current paper, we have successfully synthesized three new mercury coordination polymers with fascinating structures and properties via a flexible sulfur donor ligand, namely, {[Hg(µ2-Cl)(µ2-Ls)]}n[BF4]n(1), {[Hg(µ2-Cl)(µ2-Ls)]}n[ClO4]n(2), and [Hg(SCN)2(µ2-Ls)]n(3) [Ls = 1,1-bis(3-methyl-4-imidazoline-2-thione)methane]. These complexes have been characterized by means of different techniques such as single crystal X-ray crystallography, FT-IR, elemental analysis (CHNS), UV-Vis, PXRD, BET, and TGA. Suitable single crystals of all complexes were obtained using the branch tube method with a very high yield and good stability due to the high affinity of mercury to bind to the thione groups. The cationic moieties of polymers 1 and 2 were isostructural, with a HgCl2S2 coordination core structure. The voids of the quasi-hexagonal packing of the columnar chains were occupied by unbonded tetrahedral BF4- ions in 1 and perchlorate anions in polymer 2. Polymer 3 has a less distorted tetrahedral geometry than 1 and 2, with a HgS4 core structure. By considering the thiophilicity of mercury, a thioamide-based Ls ligand was used to detoxify Hg(II) into insoluble polymers 1-3. The results of an MTT assay for (HepG2) liver cells confirmed the excellent cytoprotective effect of this ligand against mercury. Based on IC50 calculations, their toxicity was in order of polymer 1 > polymer 2 > polymer 3. These polymers were also considered as adsorbents for the reversible removal of iodine from solution and the kinetics of the process has been studied in detail. Interestingly, all of them showed an excellent stability and high capacity, in order of 763.53 mg g-1, 877.10 mg g-1, and 905.31 mg g-1 for polymers 1-3, respectively.

Dutasteride nanoemulsion preparation to inhibit 5-alpha-hair follicle reductase enzymes in the hair follicle; an ex vivo study.

Alopecia is a treatable disorder that usually occurs due to high levels of 5-alpha dihydrotestosterone in hair follicles. To enhance the storage capacity of hair follicles and alleviate the inherent characteristics of dutasteride, 5-alpha reductase inhibitor, a prolonged-release nanocarrier was synthesised, and its influence on rat abdomen's skin was investigated. Results showed the lower ratio of S/Co (higher ethanol concentration) increased the hydrodynamic nanocarriers' particle size due to thermodynamic disturbance and Ostwald ripening. In contrast, an increase in surfactant through a decrease in interfacial tension resulted in smaller nanocarriers of 32.4 nm. Moreover, an increase in viscosity had an inverse correlation with the nanoemulsions' particle size. Nanocarriers containing ethanol showed less entrapment efficacy, perhaps due to the rapid dissolution of dutasteride into ethanol during nanoemulsification, while, based on Stokes' equation, the addition of ethanol resulted in smaller particle size and stability of the system. Skin permeation analysis using Franz diffusion cells showed nanocarriers could pass through the skin and release dutasteride for 6 days. In conclusion, the optimum concentration of ingredients is decisive in guaranteeing the ideal particle size, stability, and skin permeation of nanocarriers. The Present dutasteride nanocarrier would promise a prolonged and sustained-release drug delivery system for Alopecia therapy.

A free fatty acid receptor agonist inducing autophagy in HT-29 cells by downregulating the AKT/mTOR signaling pathway.

AIMS: GW9508, a free fatty acid receptor agonist acts in a G-coupled protein receptor 40 (GPR40)-dependent pathway. Here, we investigated the induction of stress oxidative and autophagy by GW9508 in the human colorectal cancer cell line (HT-29) and the crosstalk between autophagy and apoptotic in HT-29 cells. METHODS: HT-29 was treated with GW9508 at a concentrations range of 50-500 µM in fibrin gel. Cell viability was investigated using an MTT assay. Induction of autophagy and apoptosis was assessed through Western blotting for associated proteins, acridine orange staining, MDC staining, qRT-PCR, and electron microscopy. Also, we estimated the molecular interactions between GW9805 and some markers through molecular docking. RESULTS: GW9508 inhibited HT-29 cell proliferation, induced apoptosis, and resulted in autophagy. The induced autophagy in cells was confirmed by the observation of autophagosomes, the presence of autophagy markers, including beclin-1, LC3, AMPK, and lack expression of mTOR and AKT. Moreover, GW9508 treatment significantly increased the expression of catalase and superoxide dismutase in cells. DISCUSSION: Our results indicated that GW9508 could induce autophagy by inhibiting the Akt/mTOR in HT-29. Hence, GW9508 is suggested as a novel anticancer reagent.

Strong Binding of Phytochemicals to the Catalytic Domain of Tyrosine Hydroxylase as a Trojan Horse Decreases Dopamine in Dopaminergic Cells: Pharmaceutical Considerations in Schizophrenia and Parkinson's Disease.

BACKGROUND: Imbalances in dopamine levels result in neurological and psychological disorders such as elevated dopamine in Parkinson's disease. OBJECTIVE: Despite a considerable number of advertisements claiming Aloe-vera's effectiveness in PD treatment, it has hidden long-term disadvantages for healthy people and PD patients. METHODS: In the present investigation, the impacts of Aloe-vera on dopaminergic cells were evaluated. RESULTS: The results indicated that the focal adhesion kinase (FAK) enhancement was in line with the Bax/Bcl2 ratio decrement, reactive oxygen specious (ROS) production, and nonsignificant alteration in the sub-G1phase of the cell cycle. It led to glial cell-derived neurotrophic factor (GDNF) upregulation but did not significantly change the BDNF level involved in depression and motor impairment recovery. These events apparently resulted in the enhancement in dopaminergic cell viability and neurite length and attenuated PI+ cells. However, it also induced neuronal nitric oxide synthase (nNOS) overexpression and nitric oxide (NO) and lactate dehydrogenase (LDH) production. Notably, docking results of the catalytic domain in tyrosine hydroxylase (TH) with the Aloe-vera constituents showed strong binding of most Aloe-vera constituents with the catalytic domain of TH, even stronger than L-tyrosine as an original substrate. Following the docking results, Aloe-vera downregulated TH protein and attenuated dopamine. CONCLUSION: It can be hypothesized that Aloe-vera improves PD symptoms through enhancement in antiapoptotic markers and neurotrophic factors, while it suppresses TH and dopamine in the form of a Trojan horse, later resulting in the future deterioration of the disease symptoms. The results provide cues to pharmaceutical companies to use the active components of Aloe-vera as putative agents in neurological and psychiatric disorders and diseases to decrease dopamine in patients with enhanced dopamine levels.

Differentiation of Human Wharton's Jelly Mesenchymal Stem Cells into SOX17 Expressing Cells Using a Wnt/ß-catenin Pathway Agonist on Polylactic Acid/Chitosan Nanocomposite Scaffold.

Objective: The ß-catenin signaling pathway promises the potential for differentiation of stem cells into definitive endoderm (DE) cells as precursors of beta cells. Therefore, it can be considered as an inducer for cell replacement therapies in diabetes. The main goal of this research is to successfully culture and induce differentiation of human Wharton's jelly mesenchymal stem cells (hWJMSCs) into Sox17-expressing cells using a Wnt/ß-catenin pathway agonist (SKL2001) plus nanoparticles on a polylactic acid/chitosan (PLA/Cs) nanocomposite scaffold. Materials and Methods: In this experimental study, the nanocomposite was prepared through an electrospinning method and hWJMSCs were isolated through an explant technique. The morphology and the cell viability were evaluated by scanning electron microscopy (SEM) and 3-(4, 5- Dimethylthiazol-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. Here, we present two differentiation protocols: the first one is induction with SKL2001; and the second one is with a combination of SKL2001 and zinc oxide nanoparticles (nZnO). Real-time quantitative reverse transcription (QRT-PCR) and immunocytochemistry analysis are carried out to examine the expression of specific markers in the differentiated cells. Results: The nanocomposite had appropriate biocompatibility for cell adhesion and growth. While the hWJMSCs cultured on the PLA/Cs scaffolds differentiated into DE cells in the presence of SKL2001, introducing nZnO to their environment increased the differentiation process. Analyses of DE-specific markers including SOX17, FOXA2, and gooscoid (GSC) genes in mRNA level, indicated significantly high levels of expression in the SKL2001/nZnO group, followed by SKL2001 group compared to the control. Conclusion: Our results show the beneficial effects of the Wnt/ß-catenin pathway agonist in three-dimensional (3D) cultures in cell replacement therapy for diabetes.

Small Molecule Differentiate PDX1-Expressing Cells Derived from Human Endometrial Stem Cells on PAN Electrospun Nanofibrous Scaffold: Applications for the Treatment of Diabetes in Rat.

In this study, we designed an engineered tissue and transplanted it to an animal model, trying to take an effective step toward meeting the needs of diabetic patients. Here, human endometrial cells were differentiated into PDX1-expressing cells using a small molecule of Y-27632 on polyacrylonitrile (PAN) electrospun scaffolds and transplanted into diabetic rats. PAN nanofibers were made by electrospinning. RT-PCR and immunocytochemical analysis were performed to express pancreatic precursor (PP) genes. The differentiated cells were then transplanted into the abdominal cavity of diabetic rats with Streptozotocin. In another group of rats, differentiated cells were injected through the tail. Blood glucose was measured 7, 14, and 28 days after transplantation, and rat weight was also measured. The results showed that the expression of PP markers including Sox-17, Ngn3, Pdx1, and NKx2.2 genes was significantly increased in differentiated cells compared to the control group. In diabetic rats receiving differentiated cells, both transplanted and injected, glucose concentration as well as body weight improved compared to the control group. Rats receiving transplants in the peritoneum had a lower blood glucose concentration than those in the cell receiving group by injection, and the cell receiving group in the form of injections was more effective in increasing the body weight of rats than in the other groups. According to the results of the study, the transplantation of PP from endometrium using PAN scaffolding at the site of peritoneum could be recommended for the treatment of diabetes, although further studies are needed to provide a complete cure.

Silymarin effect on experimental bone defect repair in rat following implantation of the electrospun PLA/carbon nanotubes scaffold associated with Wharton's jelly mesenchymal stem cells.

In this study, the ability of silymarin to heal rat calvarial bone critical defects with mesenchymal stem cells isolated from human Wharton's jelly (HWJMSC) cultured on the electrospun scaffold of poly (lactic acid)/carbon nanotube (PLA/CNT) has been examined. In this study, 20 adult male Wistar rats were divided into four groups of five each. Under general anesthesia, 8 mm defects were created in the calvarial bone of the rats. Then, study groups were defined as no treatment group, the scaffold alone, the scaffold and HWJMSCs, and the scaffold/cells plus oral silymarin, respectively. The histomorphometric study was performed using H&E staining and Goldner's Masson trichrome as specific staining. The results of this study showed that the electrospun PLA/CNT scaffold is a biocompatible scaffold and HWJMSCs can considerably attach and proliferate on this scaffold, and the scaffold itself is also a suitable option for improving the bone repair process. The results of the histomorphometric analysis also showed a significantly higher amount of recently formed bone in the silymarin group plus scaffold/cells compared to the scaffold and cell group alone (p < .05). Utilizing silymarin plus HWJMSCs cultured on PLA/CNT scaffold can be used as a suitable method for the process of osteogenesis and bone repair.

Differentiation of endometrial stem cells into insulin-producing cells using signaling molecules and zinc oxide nanoparticles, and three-dimensional culture on nanofibrous scaffolds.

Diabetes mellitus is the most common metabolic disorder with a high mortality and morbidity rate. A new promising strategy to treat DM is pancreatic tissue engineering. We described a 3D culture system accompanied by signaling factors to differentiate hEnSCs into IPCs in the presence of nZnO. We isolated EnSCs and cultured it in DMEM/F12 medium. Nanofibrous PLA/Cs scaffold was prepared through the electrospinning method. The morphological properties of the scaffolds and cells were evaluated by SEM. MTT assay was used to investigate the metabolic activity of the hEnSCs cultured on the scaffolds and a four-stage protocol was applied to differentiate hEnSCs. The differentiated cells were tested for pancreatic markers by immunocytochemistry, qRT-PCR and DTZ staining. The results of this study revealed that hEnSCs cultured on PLA/Cs scaffold and treated with nZnO can efficiently differentiate into IPCs. The examination of differentiated cell morphology showed their near similarity with pancreatic islet cells, and DTZ staining emphasized the presence of insulin granules inside their cytoplasm. Moreover, qRT-PCR and immunofluorescent staining results showed the efficient expression of specific gene markers of IPCs in resultant differentiated cells. Moreover, PLA/CS and nZnO were able to provide a good nanoenvironment for the differentiation of hEnSCs into IPCS the in presence of other molecules.

Core and biological motif of self-assembling peptide nanofiber induce a stronger electrostatic interaction than BMP2 with BMP2 receptor 1A.

Recent studies suggest that nanotopography can trigger colocalization of integrins and bone morphogenetic protein 2 (BMP2) receptors (e.g., BMPR1A), thereby leading to osteogenesis. In this study, the bone marrow homing peptide 1 (BMHP1) motif was bound to a self-assembling peptide core to form a hydrogel-based nanofiber (R-BMHP1). The docking and molecular dynamic study revealed that the R-BMHP1 sequence induced a stronger electrostatic interaction than BMP2 through arginines in the RADA core sequence and through lysine24 in the BMHP1 motif with BMPR1A. Notably, decrease of polar solvation binding energy will enhance the total binding energy and increases bone regeneration even more than BMP2 The enhanced osteogenesis and bone repair potential of R-BMHP1 nanofiber might be related to its chemical interaction with BMPR1A, which triggered downstream signal transduction through osteogenic genes overexpression in osteo-differentiated mesenchymal stem cells (MSCs), as well as implanted critical-sized bone defects in rats. Following that, calcium deposition occurred by osteoblast-like cells, ALP activity increased in osteodifferentiation MSCs and rat serum, and calcium density improved in bone defects (X-ray). The nanofiber was biocompatible and enhanced the cell viability of MSCs, without multinuclear cell infiltration into the defect site. Taking everything into account, not only does nanotopography induce osteogenesis through colocalization of BMPRs and integrins, but also R-BMHP1 nanofibers (considering their chemical structure) induce cell proliferation, osteogenesis, and bone repair through strong electrostatic interaction with BMPR1A and downstream signaling. The entire outcome of this study manifests the plausibility of R-BMHP1 for spine and spinal cord injury repair.