Novel antibacterial biodegradable Fe-Mn-Ag alloys produced by mechanical alloying

Various compositions and synthesis methods of biodegradable iron-based alloys have been studied aiming for the use of temporary medical implants. However, none is focused on nano-structured alloy and on adding antibacterial property to the alloy. In this study, new Fe-30Mn-(1–3)Ag alloys were synthesized by means of mechanical alloying and assessed for their microstructure, mechanical properties, corrosion rate, antibacterial activity and cytotoxicity. Results showed that the alloy with 3 wt% Ag content displayed the highest relative density, shear strength, micro hardness and corrosion rate. However, optimum cytotoxicity and the antibacterial activity were reached by the alloy with 1 wt% Ag content. The compositional and processing effects of the alloys' properties are further discussed in this work

In vitro evaluation of human endometrial stem cell-derived osteoblast-like cells’ behavior on gelatin/collagen/bioglass nanofibers’ scaffolds

New biomimetic nanocomposite scaffold was prepared by the combination of nanofibrilar bioglass containing copper ion as the inorganic phase and gelatin/collagen as the organic phase of bone tissue. In this study for fabrication of the scaffold, freeze drying and electrospinning methods were used, and genipin was used as the cross-linking agent for increasing the mechanical properties of the scaffold. The growth and viability of human endometrial stem cell-derived osteoblast-like cells were investigated on this biomimetic scaffold. Cellular biocompatibility assays illustrated that this scaffold has more viabilities and osteoblast growths in comparison with two-dimensional culture. Copper ion increased growth of the osteoblasts on nanocomposite scaffold containing nanofibrous bioglass. Thus, the results obtained from this study indicate that the prepared scaffold is suitable for osteoblast growth and attachment; thus, potentially, this nanocomposite scaffold is an appropriate scaffold for bone tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2210–2219, 2016

Differentiation of human endometrial stem cells encapsulated in alginate hydrogel into oocyte-like cells

Introduction: Human endometrial mesenchymal stem cells (hEnMSCs) are a rich source of mesenchymal stem cells (MSCs) with multi-lineage differentiation potential, making them an intriguing tool in regenerative medicine, particularly for the treatment of reproductive and infertility issues. The specific process of germline cell-derived stem cell differentiation remains unknown, the aim is to study novel ways to achieve an effective differentiation method that produces adequate and functioning human gamete cells. Methods: We adjusted the optimum retinoic acid (RA) concentration for enhancement of germ cell-derived hEnSCs generation in 2D cell culture after 7 days in this study. Subsequently, we developed a suitable oocyte-like cell induction media including RA and bone morphogenetic protein 4 (BMP4), and studied their effects on oocyte-like cell differentiation in 2D and 3D cell culture media utilizing cells encapsulated in alginate hydrogel. Results: Our results from microscopy analysis, real-time PCR, and immunofluorescence tests revealed that 10 µM RA concentration was the optimal dose for inducing germ-like cells after 7 days. We examined the alginate hydrogel structural characteristics and integrity by rheology analysis and SEM microscope. We also demonstrated encapsulated cell viability and adhesion in the manufactured hydrogel. We propose that in 3D cell cultures in alginate hydrogel, an induction medium containing 10 µM RA and 50 ng/mL BMP4 can enhance hEnSC differentiation into oocyte-like cells. Conclusion: The production of oocyte-like cells using 3D alginate hydrogel may be viable in vitro approach for replacing gonad tissues and cells

Tissue engineering and stem cell-based therapeutic strategies for premature ovarian insufficiency

Premature ovarian insufficiency (POI), also known as premature ovarian failure (POF), is a complex endocrine disease that commonly affects women under the age of 40. It is characterized by the cessation of ovarian function before the age of 40, leading to infertility and hormonal imbalances. The currently available treatment options for POI are limited and often ineffective. Tissue engineering and stem cell-based therapeutic strategies have emerged as promising approaches to restore ovarian function and improve the quality of life for women affected by POI. This review aims to provide a comprehensive overview of the types of stem cells and biomaterials used in the treatment of POI, including their biological characteristics and mechanisms of action. It explores various sources of stem cells, including embryonic stem cells, induced pluripotent stem cells, and adult stem cells, and their potential applications in regenerating ovarian tissue. Additionally, this paper discusses the development of biomaterials and scaffolds that mimic the natural ovarian microenvironment and support the growth and maturation of ovarian cells and follicles. Furthermore, the review highlights the challenges and ethical considerations associated with tissue engineering and stem cell-based therapies for POI and proposes potential solutions to address these issues. Overall, this paper aims to provide a comprehensive overview of the current state of research in tissue engineering and stem cell-based therapeutic strategies for POI and offers insights into future directions for improving treatment outcomes in this debilitating condition

Differentiation of endometrial stem cells into motor neurons by the use of purmorphamin small molecule

Background: Small molecule Purmorphamin (PMA) is the agonist of smoothened protein in Sonic hedgehog (Shh) signaling pathway. Effect of purmorphamin small molecule on differentiation of mesenchymal cells into bone tissue has been studied previously. Use of Shh causes progression of neural differentiation, and the differentiated cells express specific neural markers. Neurofilament (NF) and acetylcholine esterase (Chat) are specific markers of motor neurons and their expression in differentiated cells indicates their conversion into motor neurons. The aim of this study was to evaluate the ability of PMA to differentiate the human endometrial stem cells (hEnSCs) into motor neurons. Methods: This analytical study was done in Tehran University of Medical Sciences laboratory on September of 2015. In this study hEnSCs were enzymatically extracted from endometrial tissue. After third passages, the flow cytometry was done for mesenchymal stem cells markers. The mesenchymal stem cells were divided into control and differentiated groups. FBS 10%+DMEM/F12 was added to the culture medium of control group and the differentiating group was treated with differentiating medium containing N2, PMA, DMEM/F12, FBS, B27, IBMX, 2ME, FGF2, RA, BDNF. After 21 days immunocytochemistry (ICC) test was done for the expression of NF and Chat proteins and Real-time PCR analysis for expression of neural markers such as NF, Chat, Nestin and GFAP (as glial marker) at mRNA level. Results: The flow cytometry analysis showed that hEnSCs were positive for mesenchymal markers CD90, CD105 and CD146 and negative for endothelial marker CD31, and hematopoietic marker CD34. The immunocytochemistry and Real time-PCR results showed that the cells treated with PMA expressed motor neuron markers of NF and Chat. Conclusion: According to the results of this study, it can be concluded that small molecule PMA has the potency to induce the differentiation of hEnSCs into neural cells, specifically motor neurons by activating Shh signaling pathway

In vitro degradation, hemocompatibility and cytocompatibility of nanostructured absorbable Fe-Mn-Ag alloys for biomedical application

The addition of noble elements such as Ag was shown as a successful method to accelerate the corrosion rate of absorbable Fe-based alloys. One major concern of Ag addition is its effect on hemocompatibility and biocompatibility. In this study, in vitro degradation and surface analysis of Fe-30Mn-xAg (x= 0, 1 and 3 wt.%) alloys as well as their effects on hemocompatibility and cell viability of human umbilical vein endothelial cells (HUVECs) were investigated. The static degradation rate of the alloys was 4.97, 4.69 and 4.49 mg/cm2 for Fe-30Mn, Fe-30Mn-1Ag, and Fe-30Mn-3Ag, respectively. The surface analysis after degradation showed that γ-FeOOH was formed on Fe-30Mn-3Ag, while α-FeOOH was more dominant on Fe-30Mn and Fe-30Mn1Ag. As γ-FeOOH is more soluble than α-FeOOH, it assists further degradation of Fe-30Mn3Ag alloy. The high amount of Ag, induced hemolysis ratio however, inhibited coagulation by decreasing the platelet adhesion. Fe-30Mn-1Ag and Fe-30Mn-3Ag alloys shown improved cell viability compared to that of Fe-Mn alloy. Shear yield strength and shear elastic modulus of the samples after immersion tests were increased while the ultimate shear strength was not affected. Based on acceptable hemolysis rate, low platelet adhesion, acceptable cell viability, and appropriate mechanical properties after degradation Fe-30Mn-1Ag can be considered as a suitable blood-contacting Fe-based absorbable alloy

Exploring the various effects of Cu doping in hydroxyapatite nanoparticle

Abstract Adding foreign ions to hydroxyapatite (HAp) is a popular approach for improving its properties. This study focuses on the effects of calcium substitution with copper in HAp. Instead of calcium, copper ions were doped into the structure of hydroxyapatite nanoparticles at 1%, 3%, and 5% concentrations. XRD analysis showed that the amount of substituted copper was less than needed to generate a distinct phase, yet its lattice parameters and crystallinity slightly decreased. Further, the results of degradation tests revealed that copper doping in hydroxyapatite doubled calcium ion release in water. The incorporation of copper into the apatite structure also boosted the HAp zeta potential and FBS protein adsorption onto powders. According to antibacterial investigations, a concentration of 200 mg/ml of hydroxyapatite containing 5% copper was sufficient to effectively eradicate E. coli and S. aureus bacteria. Furthermore, copper improved hydroxyapatite biocompatibility. Alkaline phosphatase activity and alizarin red tests showed that copper in hydroxyapatite did not inhibit stem cell differentiation into osteoblasts. Also, the scratch test demonstrated that copper-containing hydroxyapatite extract increased HUVEC cell migration. Overall, our findings demonstrated the utility of incorporating copper into the structure of hydroxyapatite from several perspectives, including the induction of antibacterial characteristics, biocompatibility, and angiogenesis

Preparation of bilayer tissue-engineered polyurethane/poly-L-lactic acid nerve conduits and their in vitro characterization for use in peripheral nerve regeneration

Abstract Background Due to loss of peripheral nerve structure and/or function resulting from trauma, accidents, and other causes, peripheral nerve injuries continue to be a major clinical problem. These injuries can cause partial or total loss of sensory, motor, and autonomic capabilities as well as neuropathic pain. PNI affects between 13 and 23 out of every 100,000 people annually in developed countries. Regeneration of damaged nerves and restoration of function after peripheral nerve injury remain significant therapeutic challenges. Although autologous nerve graft transplantation is a viable therapy option in several clinical conditions, donor site morbidity and a lack of donor tissue often hinder full functional recovery. Biomimetic conduits used in tissue engineering to encourage and direct peripheral nerve regeneration by providing a suitable microenvironment for nerve ingrowth are only one example of the cutting-edge methods made possible by this field. Many innate extracellular matrix (ECM) structures of different tissues can be successfully mimicked by nanofibrous scaffolds. Nanofibrous scaffolds can closely mimic the surface structure and morphology of native ECMs of many tissues. Methods In this study, we have produced bilayer nanofibrous nerve conduit based on poly-lactic acid/polyurethane/multiwall carbon nanotube (PLA/PU/MWCNT), for application as composite scaffolds for static nerve tissue engineering. The contact angle was indicated to show the hydrophilicity properties of electrospun nanofibers. The SEM images were analyzed to determine the fiber’s diameters, scaffold morphology, and endometrial stem cell adhesion. Moreover, MTT assay and DAPI staining were used to show the viability and proliferation of endometrial stem cells. Results The constructed bilayer PLA/PU/MWCNT scaffolds demonstrated the capacity to support cell attachment, and the vitality of samples was assessed using SEM, MTT assay, and DAPI staining technique. Conclusions According to an in vitro study, electrospun bilayer PLA/PU/MWCNT scaffolds can encourage the adhesion and proliferation of human endometrial stem cells (hEnSCs) and create the ideal environment for increasing cell survival

Application of Platelet Rich Fibrin in Tissue Engineering: Focus on Bone Regeneration

Bone tissue engineering (BTE) is a strategy for reconstructing bone lesions, which is rapidly developing in response to higher demands for bone repairing. Recently, this method, along with the emergence of functionally graded, biocompatible and biodegradable materials, has been expanded. Moreover, scaffolds with chemical, physical and external patterns have induced bone regeneration. However, the maintenance of healthy bone and its regeneration in the human body needs a series of complex and accurate processes. Hence, many studies have been accompanied for reconstructing bone by using blood-derived biomaterials, especially platelet-rich fabricates. The most important reason for using platelet-rich formulations in bone regeneration is based on releasing growth factors from alpha granules in platelets, which can induce osteogenesis. Moreover, the presence of fibrin nano-fiber structures as a constituent can provide a good substrate for cell attachments. This study attempts to review the history, structure, and biology of platelet-rich fibrin (PRF) as well as in vitro, pre-clinical, and clinical studies on the use of PRF for bone regeneration

Differentiation of human endometrial stem cells into endothelial-like cells on gelatin/chitosan/bioglass nanofibrous scaffolds

The capacity of gelatin/chitosan/bioactive glass nanopowders (GEL/CS/BGNPs) scaffolds was investigated for increasing human endometrial stem cells (hEnSCs) differentiation into the endothelial cells in the presence of angiogenic factors. GEL/CS nanofibrous scaffold with different contents of BGNPs were fabricated and assessed. Expression of endothelial markers (CD31, vascular endothelial cadherin (VE-cadherin), and KDR) in differentiated cells was evaluated. Results showed the diameter of nanofiber increases with decreasing