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OBJECTIVE: The objective of this study was to compare culture-expanded bone marrow-derived mesenchymal stem cell and PRP loaded to natural bone mineral (Bio-Oss) and beta-TCP for rat calvarial bone repair. METHODS: Twenty-two adult male Sprague-Dawley rats were randomly divided into 2 groups (n = 11). In the first group, 2 calvarial defects, 5 mm in diameter, were prepared in the parietal bone. The left defect was filled with Bio-Oss plus PRP while the right defect was filled with mesenchymal stem cell cultured on Bio-Oss. Kasios as a bone substitute was replaced in the second group. The animals were humanely killed 6 weeks postsurgery and the amount of the bone regeneration evaluated using histometric analysis. RESULT: The bone fill length in the calvarial defect had statistically significant difference with other groups (P < .05). In both groups, mesenchymal stem cell revealed a higher percentage of bone formation in comparison with the PRP groups. CONCLUSIONS: New synthetic bone substitutes may offer a better condition for bone regeneration compared to the traditional bone substitute in combination with mesenchymal stem cells. They remained in the defect and contributed bone regeneration. The use of the mesenchymal stem cell in conjunction with bone substitutes can enhance bone regeneration more than PRP. [ABSTRACT FROM AUTHOR]

In adult mammals, limb regeneration is limited by the absence of blastemal cells (BCs) and the lack of the regenerative signaling cascade. The utilization of transgenic cells circumvents the limitations associated with the absence of BCs. In a previous investigation, we successfully regenerated mouse phalanx amputations using blastema-like cells (BlCs) generated from bone marrow-derived mesenchymal stem cells (mBMSCs) overexpressing Msx1 and Msx2 genes. Recently, extracellular vesicles (EVs) have emerged as potent biological tools, offering a promising alternative to manipulated cells for clinical applications. This research focuses on utilizing BlCs-derived extracellular vesicles (BlCs-EVs) for regenerating mouse digit tips. The BlCs were cultured and expanded, and then EVs were isolated via ultracentrifugation. The size, morphology, and CD81 marker expression of the EVs were confirmed through Dynamic Light Scattering (DLS), Scanning Electron Microscope (SEM), and Western Blot (WB) analyses. Additionally, WB analysis demonstrated the presence of MSX1, MSX2, FGF8, and BMP4 proteins. The uptake of EVs by mBMSCs was shown through immunostaining. Effects on cell proliferation, migration, and osteogenic activity post-treatment with BlCs-EVs were assessed through MTT assay, scratch assay, and Real-time PCR. The regenerative potential of BlCs-EVs was evaluated in a mouse digit tip amputation model using histological assessments. Results indicated that BlCs-EVs enhanced several abilities of mBMSCs, such as migration, proliferation, and osteogenesis in vitro. Notably, BlCs-EVs significantly improved digit tip regeneration in mice, promoting the formation of new bone and nails, which was absent in control groups. In summary, BlCs-EVs are promising tools for digit tip regeneration, avoiding the ethical concerns associated with using genetically modified cells., (© 2024. The Author(s).)

This review paper explores the cutting-edge advancements in hydrogel design for articular cartilage regeneration (CR). Articular cartilage (AC) defects are a common occurrence worldwide that can lead to joint breakdown at a later stage of the disease, necessitating immediate intervention to prevent progressive degeneration of cartilage. Decades of research into the biomedical applications of hydrogels have revealed their tremendous potential, particularly in soft tissue engineering, including CR. Hydrogels are highly tunable and can be designed to meet the key criteria needed for a template in CR. This paper aims to identify those criteria, including the hydrogel components, mechanical properties, biodegradability, structural design, and integration capability with the adjacent native tissue and delves into the benefits that CR can obtain through appropriate design. Stratified-structural hydrogels that emulate the native cartilage structure, as well as the impact of environmental stimuli on the regeneration outcome, have also been discussed. By examining recent advances and emerging techniques, this paper offers valuable insights into developing effective hydrogel-based therapies for AC repair., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Rheumatoid arthritis (RA) remains a challenging chronic autoimmune disorder characterized by persistent joint inflammation and damage. While modern regenerative strategies, encompassing cell/stem cell-based therapies, gene therapy, and tissue engineering, have advanced tissue repair efforts, a definitive cure for RA remains elusive. Consequently, there is growing interest in developing targeted therapies that directly address the underlying mechanisms driving RA pathogenesis, such as extracellular vesicles (EVs). These small membrane-bound particles can modulate immune responses within the inflammatory microenvironment of damaged cartilage. To launch the clinical potential of EVs, they can be isolated from various cell types through several techniques. EVs can carry various bioactive molecules and anti-inflammatory or pro-regenerative drugs, deliver them directly to the affected joints, and affect the behavior of injured cells, making them a compelling choice for targeted therapy and drug delivery in RA patients. However, there are still several challenges and limitations associated with EV-based therapy, including the absence of standardized protocols for EV isolation, characterization, and delivery. This review provides a comprehensive overview of the cellular sources of EVs in RA and delves into their therapeutic potential and the hurdles they must overcome., (© 2024. The Author(s).)

Rheumatoid arthritis (RA) is a chronic and systemic autoimmune disease of unknown etiology. The most common form of this disease is chronic inflammatory arthritis, which begins with inflammation of the synovial membrane of the affected joints and eventually leads to disability of the affected limb. Despite significant advances in RA pharmaceutical therapies and the availability of a variety of medicines on the market, none of the available medicinal therapies has been able to completely cure the disease. In addition, a significant percentage (30-40%) of patients do not respond appropriately to any of the available medicines. Recently, mesenchymal stromal cells (MSCs) have shown promising results in controlling inflammatory and autoimmune diseases, including RA. Experimental studies and clinical trials have demonstrated the high power of MSCs in modulating the immune system. In this article, we first examine the mechanism of RA disease, the role of cytokines and existing medicinal therapies. We then discuss the immunomodulatory function of MSCs from different perspectives. Our understanding of how MSCs work in suppressing the immune system will lead to better utilization of these cells as a promising tool in the treatment of autoimmune diseases., (© 2023. The Author(s).)

The purpose of cartilage tissue engineering is to provide artificial constructs with biological functions and mechanical features that resemble native tissue to improve tissue regeneration. Biochemical characteristics of the cartilage extracellular matrix (ECM) microenvironment provide a platform for researchers to develop biomimetic materials for optimal tissue repair. Due to the structural similarity of polysaccharides into physicochemical characteristics of cartilage ECM, these natural polymers capture special attention for developing biomimetic materials. The mechanical properties of constructs play a crucial influence in load-bearing cartilage tissues. Moreover, the addition of appropriate bioactive molecules to these constructs can promote chondrogenesis. Here, we discuss polysaccharide-based constructs that can be used to create substitutes for cartilage regeneration. We intend to focus on newly developed bioinspired materials, fine-tuning the mechanical properties of constructs, the design of carriers loaded by chondroinductive agents, and development of appropriate bioinks as a bioprinting approach for cartilage regeneration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Utilizing both medium enrichment and a thermos-responsive substrate to maintain the cell-to-cell junctions and extracellular matrix (ECM) intact, cell sheet technology has emerged as a ground-breaking approach. Investigating the possibility of using sodium selenite (as medium supplementation) and PCL-PEG-PCL (as vessel coating substrate) in the formation of the sheets from rat bone marrow-derived mesenchymal stem cells (rBMSCs) was the main goal of the present study. To this end, first, Polycaprolactone-co-Poly (ethylene glycol)-co-Polycaprolactone triblock copolymer (PCEC) was prepared by ring-opening copolymerization method and characterized by FTIR, 1  H NMR, and GPC. The sol-gel-sol phase transition temperature of the PCEC aqueous solutions with various concentrations was either measured. Next, rBMSCs were cultured on the PCEC, and let be expanded in five different media containing vitamin C (50 µg/ml), sodium selenite (0.1 µM), vitamin C and sodium selenite (50 µg/ml + 0.1 µM), Trolox, and routine medium. The proliferation of the cells exposed to each material was evaluated. Produced cell sheets were harvested from the polymer surface by temperature reduction and phenotypically analyzed via an inverted microscope, hematoxylin and eosin (H&E) staining, and field emission scanning electron microscopy (FESEM). Through the molecular level, the expression of the stemness-related genes (Sox2, Oct-4, Nanog), selenium-dependent enzymes (TRX, GPX-1), and aging regulator gene (Sirt1) were measured by q RT-PCR. Senescence in cell sheets was checked by beta-galactosidase assay. The results declared the improved ability of the rBMSCs for osteogenesis and adipogenesis in the presence of antioxidants vitamin C, sodium selenite, and Trolox in growth media. The data indicated that in the presence of vitamin C and sodium selenite, the quality of the cell sheet was risen by reducing the number of senescent cells and high transcription of the stemness genes. Monolayers produced by sodium selenite was in higher-quality than the ones produced by vitamin C., (© 2023. The Author(s).)

Nanocomposite hydrogels based on tyramine conjugated gum tragacanth, poly (vinyl alcohol) (PVA), and halloysite nanotubes (HNTs) were prepared by electron beam irradiation and characterized. The FTIR, 1 H NMR, and TGA results confirmed the chemical incorporation of HNTs into gum tragacanth. Gel content and swelling of hydrogels decreased with HNTs loading up to 20 % wt. The mechanical strength of hydrogels increased by increasing HNTs content up to 10 % with 371 kPa fracture stress at 0.95 fracture strain, compared to 312 kPa stress at 0.79 strain for gum tragacanth/PVA hydrogel. Hydrogel's biocompatibility and osteogenic activity were tested by seeding rabbit bone marrow mesenchymal stem cells. The cell viability was >85 % after 7 days of culture. In vitro secretion of ALP and calcium deposition on hydrogels in alizarin red assay after 21 days of culture indicated hydrogel potential for bone tissue engineering., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ebrahim Vasheghani-Farahani reports financial support was provided by Iran National Science Foundation (INSF) with grant number 96004459., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Despite advances in bone tissue engineering, fabricating a scaffold which can be used as an implant for large bone defects remains challenge. One of the great importance in fabricating a biomimetic bone implant is considering the possibility of the integration of the structure and function of implants with hierarchical structure of bone. Herein, we propose a method to mimic the structural unit of compact bone, osteon, with spatial pattern of human umbilical vein endothelial cells (HUVECs) and human mesenchymal stem cells (hMSCs) in the adjacent layers that mimic Haversian canal and lamella, respectively. To this end, coaxial extrusion-based bioprinting technique via a customized quadruple-layer core-shell nozzle was employed. 3D implant scaffold-cell construct was fabricated by using polyethylene glycol as a hollowing agent in the first layer, gelatin methacryloyl (GelMA) and alginate blended hydrogel encapsulating HUVEC cells with vascular endothelial growth factor nanoparticles in the second layer (vasculogenic layer) to mimic vascular vessel, and GelMA and alginate blended hydrogel containing hMSCs cells in the outer osteogenic layer to imitate lamella. Two types of bone minerals, whitlockite and hydroxyapatite, were incorporated in osteogenic layer to induce osteoblastic differentiation and enhance mechanical properties (the young's modules of nanocomposite increased from 35 kPa to 80 kPa). In-vitro evaluations demonstrated high cell viability (94 % within 10 days) and proliferation. Furthermore, ALP enzyme activity increased considerably within 2 weeks and mineralized extra cellular matrix considerably produced within 3 weeks. Also, a significant increase in osteogenic markers was observed indicating the presence of differentiated osteoblast cells. Therefore, the work indicates the potential of single step 3D bioprinting process to fabricate biomimetic osteons to use as bone grafts for regeneration., Competing Interests: Conflicts of interest Hae Lin Jang is a founder and owns equity in Curer Inc., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease of unknown cause. The interaction of immune system cells and the secretion of inflammatory cytokines with synovial cells leads to severe inflammation in the affected joints. Currently, medications, including non-steroidal anti-inflammatory drugs, glucocorticoids, and more recently, disease-modifying anti-rheumatic drugs, are used to reduce inflammation. However, long-term use of these drugs causes adverse effects or resistance in a considerable number of RA patients. Recent findings revealed the safety and efficacy of mesenchymal stromal cells (MSCs)-based therapies both in RA animal models and clinical trials. Here, the beneficial effects of bone marrow-derived heterogeneous MSCs (BM-hMSCs) and Wharton jelly-derived MSCs (WJ-MSCs) at early passages were compared to BM-derived clonal MSCs (BM-cMSCs) at high passage number on a rat model of collagen-induced arthritis. Results showed that systemic delivery of MSCs significantly reversed adverse changes in body weight, paw swelling, and arthritis score in all MSC-treated groups. Radiological images and histological evaluation demonstrated the therapeutic effects of MSCs. There was a decrease in serum level of anti-collagen type II immunoglobulin G and the inflammatory cytokines interleukin (IL)-1β, IL-6, IL-17, and tumor necrosis factor-α in all MSC-treated groups. In contrast, an increase in inhibitory cytokines transforming growth factor-β and IL-10 was seen. Notably, the long-term passages of BM-cMSCs could alleviate RA symptoms similar to the early passages of WJ-MSCs and BM-hMSCs. The importance of BM-cMSCs is the potential to establish cell banks with billions of cells derived from a single donor that could be a competitive cell-based therapy to treat RA., (© 2022 John Wiley & Sons Ltd.)

Conventional treatments for osteoarthritis (OA), including drug delivery and tissue engineering approaches, could not offer a high yield of cartilage repair due to the compact and exclusive structure of cartilage. Targeted and high-efficiency delivery of gene sequences is necessary to rebalance the lost homeostatic properties of the cartilage in OA. Herein, we synthesized chitosan (CH)-chondroitin sulfate (CS) nanoparticles (NPs) as a platform for delivering gene sequences. These new nanoparticles benefit from two natural polymers that minimize the toxicity, and the presence of CS can be in favor of targeted delivery. The CAG-GFP plasmid was used as a gene sequence model, and the nanoparticles could successfully encapsulate approximately all of them in their structure. Loaded nanoparticles were characterized in terms of morphology, size, zeta potential, the efficiency of encapsulation and, DNA release pattern. Cell viability and uptake of new nanoparticles were compared to the chitosan nanoparticles and Lipofectamine. After substituting TPP with CS, NPs exhibited a significant decrease in size. In addition, there was little difference in zeta potential between nanoparticles. Furthermore, a tremendous increase in plasmid uptake and cell viability was observed by CH-CS NPs compared to CH-TPP NPs and Lipofectamine. In the final stage, the knockdown level of MMP13 was evaluated with real-time RT-PCR for confirming the potential uptake of CH-CS NPs. The results revealed cellular uptake of siRNA loaded NPs and effective knockdown of MMP13 in chondrocytes. In conclusion, CH-CS nanoparticles can be considered as a candidate for gene therapy purposes in cartilage diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Recently, postoperative bone infections have been one of the most crucial challenges for surgeons. This study aims to synergistically promote antibacterial and osteoconductive properties of hydroxyapatite (HAp) nanoparticles through binary doping of Zn 2+ and Ga 3+ ions (Zn-Ga:HAp). Zn-Ga:HAp nanopowders with spherical morphology and homogeneous size are synthesized using a simple sol-gel method. Substitution of both zinc and gallium in the structure of HAp results in a gradual decrease in the lattice parameters as doping level increases, limits the growth of HAp particles and reduces its crystallinity. Noticeably, the crystallinity of HAp (85%) reduces to less than 73% (for X Zn = 0.1), 78% (for X Ga = 0.4) and 75% (for X Zn = 0.1 and X Ga = 0.4). Ion doping also significantly modulate the release of bioactive ions (Ca 2+ , PO 4 3- , Zn 2+ , Ga 3+ ) from the Zn-Ga:HAp depended on the overall amount of Ga and Zn in the HAp, which could mediate the biological responses. Incorporating both Zn 2+ and Ga 3+ ions in HAp structure could significantly improve the antibacterial activity of HAp nanopowders against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) with a concentration-dependent effect. Noticeably, Zn-Ga:HAp (X Zn = 0.1 and X Ga = 0.4) powder shows the antibacterial activity of more than 68% and 84% against E. coli and S. aureus, respectively, at the concentration of 500 μg/ml, thereby showing excellent antibacterial properties. In addition, Zn-Ga:HAp nanopowders not only do not exhibit any cytotoxicity towards hMSCs, but also show significantly superior osteogenic properties. For instance, Zn-Ga:HAp (X Zn = 0.1 and X Ga = 0.4) nanopowders significantly enhance the alkaline phosphatase activity (approximately 2-fold) and mineralization (approximately 3-fold) of hMSCs after 14 days of culture, compared to pure HAp. Overall, Zn-Ga:HAp (X Zn = 0.1 and X Ga = 0.4) with desired osteogenesis and antibacterial activity compared to pure HAp, Zn:HAp and Ga:HAp shows promising opportunities for the implant-associated infections and the efficient healing of bone defects., Competing Interests: Declaration of competing interest, (Copyright © 2022 Elsevier B.V. All rights reserved.)

Osteoarthritis (OA) is a common progressively degenerative joint disease that affects more than 300 million people worldwide. OA is manifested by articular cartilage degradation, chronic pain, deformity, functional disability, and decreased quality of life. A real challenge in OA management is the lack of an effective cure because exiting therapeutics often provide symptom control rather than disease modification; therefore, they fail to prevent disease progression. The inadequate treatments for OA management have encouraged researchers to study mesenchymal stem cells (MSCs) as an investigational treatment for OA. MSCs are a promising tool for OA because of their availability; expand cultivation and multi-lineage differentiation capacity as well as well-documented paracrine function have made MSCs a promising tool in this field. Accordingly, MSCs application has been successfully utilized in a broad range of pre-clinical OA animal models as well as clinical studies with the aim of cartilage repair which had not previously been achieved using classical treatments. Here, the brief scientific review of MSC role in the control of OA as well as the proposed mechanisms are discussed. We provide an insight into the last 10 years' studies conducted on preclinical and clinical OA treatment as well as future opportunities in OA management strategies employing MSCs., (© 2021. Springer Nature Switzerland AG.)

Osteoarthritis (OA) and other degenerative joint diseases are characterized by articular cartilage destruction, synovial inflammation, sclerosis of subchondral bone, and loss of extracellular matrix (ECM). Worldwide, these diseases are major causes of disability. Cell therapies have been considered to be the best therapeutic strategies for long-term treatment of articular cartilage diseases. It has been suggested that the mechanism of stem cell-based therapy is related to paracrine secretion of extracellular vesicles (EVs), which are recognized as the main secretion factors of stem cells. EVs, and in particular the subclass exosomes (Exos), are novel therapeutic approaches for treatment of cartilage lesions and OA. The results of recent studies have shown that EVs isolated from mesenchymal stem cells (MSCs) could inhibit OA progression. EVs isolated from various stem cell sources, such as MSCs, may contribute to tissue regeneration of the limbs, skin, heart, and other tissues. Here, we summarize recent findings of preclinical and clinical studies on different MSC-derived EVs and their effectiveness as a treatment for damaged cartilage. The Exos isolation techniques in OA treatment are also highlighted., (© 2021. Springer Nature Switzerland AG.)

Objectives: Leishmania major (L. major) is a cutaneous leishmaniasis causative agent. Current chemotherapeutic methods are not totally effective in treatment of this disease. The immunomodulation and tissue repairing capability of mesenchymal stem cells (MSCs), ease of isolation, detection and in vitro culture, have encouraged biologists to use MSCs for cell therapy in different infections such as cutaneous leishmaniasis., Methods: BALB/c mice (6-8 weeks old) were infected with L. major then divided into four groups and treated with MSCs, Glucantime, Glucantime + MSCs, or PBS. Regression of lesions, potency of macrophages for phagocytosis, proliferation of immune cells against Leishmania soluble antigen, reduction of spleen parasite burden and healing of the lesions were evaluated on days 10, 20 and 30 of treatment., Results: The results indicated that the mice intralesionally injected with MSCs showed significant regression in the lesions produced by L. major by day 30. Proliferation of splenocytes stimulated with SLA (soluble leishmania antigen) in vitro in MSC-treated mice on day 20 was significantly higher than in the other groups. The potency of phagocytosis in macrophages of mice treated with MSCs was significantly higher by day 30 and healing of the lesions in this group of mice showed more progress on histopathological examinations. Spleen parasite burden showed significant reduction in the mice treated with Glucantime + MSCs by day 30., Conclusions: The results showed that including MSCs in treatment of cutaneous leishmaniasis caused by L. major is a promising approach., (Copyright © 2020. Published by Elsevier Ltd.)

Background: Adult bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent stem cells that can differentiate into three lineages. They are suitable sources for cell-based therapy and regenerative medicine applications. This study aims to evaluate the hub genes and key pathways of differentially expressed genes (DEGs) related to osteogenesis by bioinformatics analysis in three different days. The DEGs were derived from the three different days compared with day 0., Results: Gene expression profiles of GSE37558 were obtained from the Gene Expression Omnibus (GEO) database. A total of 4076 DEGs were acquired on days 8, 12, and 25. Gene ontology (GO) enrichment analysis showed that the non-canonical Wnt signaling pathway and lipopolysaccharide (LPS)-mediated signaling pathway were commonly upregulated DEGs for all 3 days. KEGG pathway analysis indicated that the PI3K-Akt and focal adhesion were also commonly upregulated DEGs for all 3 days. Ten hub genes were identified by CytoHubba on days 8, 12, and 25. Then, we focused on the association of these hub genes with the Wnt pathways that had been enriched from the protein-protein interaction (PPI) by the Cytoscape plugin MCODE., Conclusions: These findings suggested further insights into the roles of the PI3K/AKT and Wnt pathways and their association with osteogenesis. In addition, the stem cell microenvironment via growth factors, extracellular matrix (ECM), IGF1, IGF2, LPS, and Wnt most likely affect osteogenesis by PI3K/AKT.

Background: Regeneration of articular cartilage poses a tremendous challenge due to its limited self-repair capability and inflammation at the damaged site. To generate the desired structures that mimic the structure of native tissue, microtissues with repeated functional units such as cell aggregates have been developed. Multicellular aggregates of mesenchymal stem cells (MSCs) can be used as microscale building blocks of cartilage due to their potential for cell-cell contact, cell proliferation, and differentiation., Methods: Chondrogenic microtissues were developed through incorporation of kartogenin-releasing poly (lactic-co-glycolic acid) (PLGA) microparticles (KGN-MP) within the MSC aggregates. The chondrogenic potential of KGN-MP treated MSC aggregates was proven in vitro by studying the chondrogenic markers at the RNA level and histological analysis. In order to address the inflammatory responses at the defect site, the microtissues were delivered in vivo via an injectable, anti-inflammatory hydrogel that contained gelatin methacryloyl (GelMA) loaded with curcumin (Cur)., Results: The KGN-MPs were fabricated to support MSCs during cartilage differentiation. According to real-time RT-PCR analysis, the presence of KGN in the aggregates led to the expression of cartilage markers by the MSCs. Both toluidine blue (TB) and safranin O (SO) staining demonstrated homogeneous glycosaminoglycan production throughout the KGN-MP incorporated MSC aggregates. The curcumin treatment efficiently reduced the expressions of hypertrophy markers by MSCs in vitro. The in vivo results showed that implantation of chondrogenic microtissues (KGN-MP incorporated MSC aggregates) using the curcumin loaded GelMA hydrogel resulted in cartilage tissue regeneration that had characteristic features close to the natural hyaline cartilage according to observational and histological results., Conclusions: The use of this novel construct that contained chondrogenic cell blocks and curcumin is highly desired for cartilage regeneration.

Objectives: We aim to develop a 3D-bilayer collagen (COL) membrane reinforced with nano beta-tricalcium-phosphate (nβ-TCP) particles and to evaluate its bone regeneration in combination with leukocyte-platelet-rich fibrin (L-PRF) in vivo., Background Data: L-PRF has exhibited promising results as a cell carrier in bone regeneration in a number of clinical studies, however there are some studies that did not confirm the positive results of L-PRF application., Methods: Mechanical & physiochemical characteristics of the COL/nβ-TCP membrane (1/2 & 1/4) were tested. Proliferation and osteogenic differentiation of seeded cells on bilayer collagen/nβ-TCP thick membrane was examined. Then, critical-sized calvarial defects in 8 white New Zealand rabbits were filled with either Col, Col/nβ-TCP, Col/nβ-TCP combined with L-PRF membrane, or left empty. New bone formation (NBF) was measured histomorphometrically 4 & 8 weeks postoperatively., Results: Compressive modulus increases while porosity decreases with higher β-TCP concentrations. Mechanical properties improve, with 89 % porosity (pore size ∼100 μm) in the bilayer-collagen/nβ-TCP membrane. The bilayer design also enhances the proliferation and ALP activity. In vivo study shows no significant difference among test groups at 4 weeks, but Col/nβ-TCP + L-PRF demonstrates more NBF compared to others (P < 0.05) after 8 weeks., Conclusion: The bilayer-collagen/nβ-TCP thick membrane shows promising physiochemical in vitro results and significant NBF, as ¾ of the defect is filled with lamellar bone when combined with L-PRF membrane., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Large bone defects treatment is one of the challenges in current bone tissue engineering approaches. Various strategies have been proposed to address this issue, among which, prevascularization by coculturing of angiogenic and osteogenic cells on the scaffolds can alleviate this problem. In the present study, modified fibrous scaffolds were prepared by electrospinning and subsequent ultrasonication of polycaprolactone (PCL) containing nano-hydroxyapatite (n-HA), with/without nano-zinc oxide (n-ZnO), and polyethylene oxide [PEO] as a sacrificial agent. The physical, mechanical, and chemical characteristics of the scaffolds were evaluated. The results showed the presence of n-ZnO, which in turn increased Young's module of the scaffolds from 5.5 ± 0.67 to 6.7 ± 1.77 MPa. Moreover, MTT, SEM, alkaline phosphatase (ALP) activity, chicken embryo chorioallantoic membrane (CAM) assay, and real-time RT-PCR were utilized to investigate the biocompatibility, cell adhesion and infiltration, osteoconductivity, angiogenic properties, and expression of osteogenic and angiogenic related genes. ALP assay showed that the highest enzyme activity was noted when the modified scaffolds containing n-ZnO were seeded with HUVEC:hBMSC at the cell ratio of 1:5. CAM assay showed induction of angiogenesis for the scaffolds containing n-ZnO. Real-time RT-PCR results showed significant upregulation of angiogenic related genes. Thus, the scaffolds containing n-ZnO may have great potential for osteogenesis and angiogenesis in tissue engineering applications., (© 2019 Wiley Periodicals, Inc.)

A major problem in the treatment of large bone defects is the inability to provide an adequate blood supply to the implantation site. Therefore, a bone regeneration strategy that provides an adequate supply of vessels would address this need. Cobalt (Co 2+ ), because of its ability to induce hypoxia, has been used to accelerate new vessel formation. In this study, we used a freeze-drying technique to fabricate a scaffold that consisted of Co 2+ -doped calcium phosphate (CaP) [e.g., hydroxyapatite (HA)] and natural silk fiber through an optimized alternate mineralization process. The composition and structure of the scaffold were confirmed by X-ray diffraction (XRD), Fourier transform infrared (FTIR), inductively coupled plasma (ICP), and scanning electron microscope (SEM). The data showed that the scaffolds promoted differentiation of adipose-derived mesenchymal stem cells (ADSCs) toward endothelial and osteoblast linages. We observed improved angiogenesis and bone formation with the fabricated scaffolds compared with the control groups. Computed tomography (CT) scans and radiographic imaging, in addition to histology and immunohistochemical analyses, showed the presence of angiogenesis and bone regeneration after implantation of the ADSC-seeded scaffolds in a critical size calavarial bone defect in a Wistar rat model. We obtained the best in vitro and in vivo results by doping 2% Co 2+ in HA. Taken together, we propose that the Co 2+ -doped HA/silk fibroin (SF) scaffold would be a good candidate to induce angiogenesis and bone formation both in vitro and in vivo.

This study aimed at investigating the expression of osteoblast and chondrocyte-related genes in mesenchymal stem cells (MSCs), derived from rabbit adipose tissue, under mechanical vibration. The cells were placed securely on a vibrator's platform and subjected to 300 Hz of sinusoidal vibration, with a maximum amplitude of 10 μm, for 45 min per day, and for 14 consequent days, in the absence of biochemical reagents. The negative control group was placed in the conventional culture medium with no mechanical loading. The expression of osteoblast and chondrocyte-related genes was investigated using real-time polymerase chain reaction (real-time PCR). In addition, F-actin fiber structure and alignment with the help of actin filament fluorescence staining were evaluated, and the level of metabolic activity of MSCs was determined by the methyl thiazolyl tetrazolium assay. The real-time PCR study showed a significant increase of bone gene expression in differentiated cells, compared with MSCs (P < 0.05). On the other hand, the level of chondrocyte gene expression was not remarkable. Applying mechanical vibration enhanced F-actin fiber structure and made them aligned in a specific direction. It was also found that during the differentiation process, the metabolic activity of the cells increased (P < 0.05). The results of this work are in agreement with the well-accepted fact that the MSCs, in the absence of growth factors, are sensitive to low-amplitude, high-frequency vibration. Outcomes of this work can be applied in cell therapy and tissue engineering, when regulation of stem cells is required.

In the present comparative study, gelatin microspheres (GMs) were prepared by emulsification-solvent-extraction method using well-known crosslinker: glutaraldehyde (GA) and biocompatible silane-coupling agent: glycidoxypropyltrimethoxysilane (GPTMS). Crosslinking with GA was done by a definite and common procedure, while GPTMS crosslinking potency was investigated after 5, 10, 24, and 48 h synthesis periods and the fabrication method was adjusted in order for preparation of GMs with optimized morphological and compositional characteristics. The prepared GMs were then evaluated and compared as drug delivery systems for the antibiotic vancomycin (Vm). Morphological observations, FTIR, ninhydrin assay, swelling behavior evaluation and Hydrolytic degradation analysis proved successful modification of GMs and revealed that increasing synthesis time from 5 h to 24 h and 48 h, when using GPTMS as crosslinker, led to formation of morphologically-optimized GMs with highest crosslinking degree (∼50%) and the slowest hydrolytic degradation rate. Such GMs also exhibited most sustained release period of Vm. The antibacterial test results against gram-positive bacterium Staphylococcus aureus, were in accordance with the release profiles of Vm, as well. Together, GPTMS-crosslinked GMs with their preferable characteristics and known as biocompatible gelatin-siloxane hybrids, could act as proper drug delivery systems for the sustained release of the antibiotic vancomycin., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Tissue engineering aims to take advantage of the ability of undifferentiated stem cells to differentiate into multiple cell types to repair damaged tissue. Photobiomodulation uses either lasers or light-emitting diodes to promote stem cell proliferation and differentiation. The present study aimed to investigate single and dual combinations of laser wavelengths on mesenchymal stem cells (MSCs). MSCs were derived from rabbit iliac bone marrow. One control and eight laser irradiated groups were designated as Infrared (IR, 810 nm), Red (R, 660 nm), Green (G, 532 nm), Blue (B, 485 nm), IR-R, IR-B, R-G, and B-G. Irradiation was repeated daily for 21 days and cell proliferation, osseous, or cartilaginous differentiation was then measured. RT-PCR biomarkers were SOX9, aggrecan, COL 2, and COL 10 expression for cartilage and ALP, COL 1, and osteocalcin expression for bone. Cellular proliferation was increased in all irradiated groups except G. All cartilage markers were significantly increased by IR and IR-B except COL 10 which was suppressed by IR-B combination. ALP expression was highest in R and IR groups during osseous differentiation. ALP was decreased by combinations of IR with B and with R, and also by G alone. R and B-G groups showed stimulated COL 1 expression; however, COL 1 was suppressed in IR-B, IR-R, and G groups. IR significantly increased osteocalcin expression, but in B, B-G, and G groups it was reduced. Cartilage differentiation was stimulated by IR and IR-B laser irradiation. The effects of single or combined laser irradiation were not clear-cut on osseous differentiation. Stimulatory effects on osteogenesis were seen for R and IR lasers, while G laser had inhibitory effects.

In the originally published article, the name of the 3rd and 4th authors were labeled incorrectly. The correct names are Mohammadreza Baghaban Eslaminejad and Leila Taghiyar. Also, affiliation 4 has been corrected.

Malignant bone tumors, although quite rare, are one of the causes of death in children and adolescents. Surgery as a common and primary treatment for removal of virtually bone cancer cause large bone defects. Thus, restoration of hard tissues like bone and cartilage after surgical tumor resection needs efficient therapeutic approaches. Tissue engineering (TE) is a powerful approach which has provided hope for restoration, maintenance, or improvement of damaged tissues. This strategy generally supplies a three-dimensional scaffold as an active substrate to support cell recruitment, infiltration, and proliferation for neo-tissues. The scaffold mimics the natural extracellular matrix (ECM) of tissue which needs to be regenerated. The use of potent cell sources such as mesenchymal stem cells (MSCs) has also led to remarkable progresses in hard tissue regeneration. Combination of living cells and various biomaterials have continuously evolved over the past decades to improve the process of regeneration. This chapter describes various strategies used in TE and highlights recent advances in cell-loaded constructs. We herein focus on cell-based scaffold approach utilized in hard tissue engineering and parameters determining a clinically efficient outcome. Also, we attempt to identify the potential as well as shortcomings of pre-loaded scaffolds for future therapeutic applications.

Growth factors have a pivotal role in chondrogenic differentiation of stem cells. The differential effects of known growth factors involved in the maintenance and homeostasis of cartilage tissue have been previously studied in vitro. However, there are few reported researches about the interactional effects of growth factors on chondrogenic differentiation of stem cells. The aim of this study is to examine the combined effects of four key growth factors on chondrogenic differentiation of mesenchymal stem cells (MSCs). Isolated and expanded rabbit bone marrow-derived MSCs underwent chondrogenic differentiation in a micromass cell culture system that used a combination of the following growth factors: transforming growth factor beta 1 (TGF-β1), bone morphogenetic protein 2 (BMP2), parathyroid hormone related protein (PTHrP), and fibroblast growth factor 2 (FGF2) according to a defined program. The chondrogenic differentiation program was analyzed by histochemistry methods, quantitative RT-PCR (qRT-PCR), and measurement of matrix deposition of sulfated glycosaminoglycan (sGAG) and collagen content at days 16, 23, and 30. The results showed that the short-term combination of TGF-β1 and BMP-2 increased sGAG and collagen content, Alkaline phosphates (ALP) activity, and type X collagen (COL X) expression. Application of either PTHrP or FGF2 simultaneously decreased TGF-β1/BMP-2 induced hypertrophy and chondrogenic markers (at least for FGF2). However, successive application of PTHrP and FGF2 dramatically maintained the synergistic effects of TGF-β1/BMP-2 on the chondrogenic differentiation potential of MSCs and decreased unwanted hypertrophic markers. This new method can be used effectively in chondrogenic differentiation programs.

Lipopolysaccharides (LPS) from gram negative bacteria stimulate toll-like receptor 4 (TLR4) expression in immune cells. Recent reports state that bone marrow-derived cells such as mesenchymal stem cells (MSCs) also express TLR proteins. Numerous researches have studied the effect of a number of LPSs on TLR4 expression, but no data exists on the effect of LPSs from different strains of one bacterial genus on TLR4 expression. In this study, we investigate the effects of various concentrations of LPS from different Shigella strains on TLR4 expression in human bone marrow (hBM)-MSCs. At the mRNA level, we have found that untreated hBM-MSCs (control) did not express TLR4 compared to the experimental groups. Cells treated with LPS from Shigella flexneri had the highest expression of TLR4, whereas cells treated with LPS from Shigella sonnei had the lowest expression. We observed that LPSs had a dose-dependent effect on TLR4 expression in all of the treatment groups. ELISA findings for interleukin-6 secretion have confirmed mRNA expression results for all treatment groups. Hence, LPS from S. flexneri can be considered as an optimum LPS to stimulate the immune system for vaccine production against shigellosis. Also, TLR activation in hBM-MSCs can modulate their function such as homing., (Copyright © 2018. Published by Elsevier Ltd.)

Limb/digit amputation is a common event in humans caused by trauma, medical illness, or surgery. Although the loss of a digit is not lethal, it affects quality of life and imposes high costs on amputees. In recent years, the increasing interest in limb regeneration has led to enhanced scientific knowledge. However, the limited ability to develop functional limb regeneration in the clinical setting suggests that a challenging issue remains in limb regeneration. Recently, the emergence of regenerative engineering is a promising field to address this challenge and close the gap between science and clinical applications. Cell signalling and molecular mechanisms involved in the limb regeneration process have been extensively studied; however, there is still insufficient data on cell therapy and tissue engineering for limb regeneration. In this review, we intend to focus on therapeutic approaches for limb regeneration that are closely related to gene, immune, and stem cell therapies, as well as tissue engineering approaches that take into consideration the peculiar developmental properties of the limbs. In addition, we attempt to identify the challenges of these strategies for limb regeneration studies in terms of clinical settings and as a road map to accomplish the goal of functional human limb regeneration., (© 2018 John Wiley & Sons, Ltd.)

Mechanical forces throughout human mesenchymal stem cell (hMSC) spheroids (mesenspheres) play a predominant role in determining cellular functions of cell growth, proliferation, and differentiation through mechanotransductional mechanisms. Here, we introduce microparticle (MP) incorporation as a mechanical intervention method to alter tensional homeostasis of the mesensphere and explore MSC differentiation in response to MP stiffness. The microparticulate mechanoregulators with different elastic modulus (34 kPa, 0.6 MPa, and 2.2 MPa) were prepared by controlled crosslinking cell-sized microdroplets of polydimethylsiloxane (PDMS). Preparation of MP-MSC composite spheroids enabled us to study the possible effects of MPs through experimental and computational assays. Our results showed that MP incorporation selectively primed MSCs toward osteogenesis, yet hindered adipogenesis. Interestingly, this behavior depended on MP mechanics, as the spheroids that contained MPs with intermediate stiffness behaved similar to control MP-free mesenspheres with more tendencies toward chondrogenesis. However, by using the soft or stiff MPs, the MP-mesenspheres significantly showed signs of osteogenesis. This could be explained by the complex of forces which acted in the cell spheroid and, totally, provided a homeostasis situation. Incorporation of cell-sized polymer MPs as mechanoregulators of cell spheroids could be utilized as a new engineering toolkit for multicellular organoids in disease modeling and tissue engineering applications., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Electrospun nanofiber matrices are widely used as scaffolds for the regeneration of different tissues due to similarities with fibrous components of the extracellular matrix. These scaffolds could act as a substrate for inducing mechanical stimuli to cells. The main mechanical stimuli factor in nanofiber scaffolds for determining the cell behaviors is stiffness of single nanofibers. This paper especially highlights the finding that the young's modulus of single nanofibers can be obtained from aligned nanofibers matrix. It is assume that, the modulus of single nanofibers are equal to modulus of completely aligned nanofibers. However, due to difficulty of producing completely aligned nanofibers, the obtained modulus of single nanofiber wouldn't have significant value. Therefore, we propose a new mathematical model to predict the stiffness of single nanofibers from non-perfectly aligned nanofibers matrix., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Some research studies provided evidence for the differentiation capacity of adult stem cells (ASCs) into germ cells (GCs). Since the generation of GCs from stem cells (SCs) has been proposed as a potential way for treatment of infertility, many research groups have begun their creative studies on generation of new GCs both in vitro and in vivo, and utilized different ASC types such as bone marrow mesenchymal stem cells (BM-MSCs), skin stem cells, pancreatic stem cells, and adipose tissue MSCs. Despite many interesting reports with promising results, an obvious problem in the research projects was the functionality of the produced GCs., Objective: In this paper, we have reviewed the results of almost all previously published reports on derivation of male and female GCs from ASCs to provide a better insight into this field of research., Results: The most evaluated papers have shown that ASCs from various tissues can differentiate into GCs but rarely were the produced GCs functional and could form fertile gametes neither in vitro, nor in vivo (after transplantation into the gonads)., Conclusion: There are still so many unknown issues about gametogenesis. Perhaps making alterations in treatment methods and utilizing creative techniques like tissue engineering and gene targeting help to achieve a standard method of in vitro GC production from ASCs., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

Biphasic calcium phosphate (BCP) microspheres are of great interest due to their high stability and osteoinductive properties at specific compositions. However, the need for optimal performance at a unique composition limits their flexibility for tuning drug release by modulation of bulk properties and presents the question of engineering surface topography as an alternative. It is necessary to have a facile method to control surface topography at a defined bulk composition. Here, we have produced BCP microspheres with different surface topographies that have the capability to be used as tunable drug release systems. We synthesized calcium deficient hydroxyapatite (CDHA) microparticles by precipitating calcium and phosphate ions onto ethylenediaminetetraacetic acid (EDTA) templates. The morphology and surface topography of CDHA microparticles were controlled using process parameters, which governed nucleation and growth. These parameters included template concentration, heat rate, and stirring speed. Under low heat rate and static conditions, we could obtain spherical microparticles with long and short nanosheets on their surfaces at low and high EDTA concentrations, respectively. These nanostructured microspheres were subsequently crystallized by thermal treatment to produce EDTA-free BCP microspheres with intact morphology. These biocompatible BCP microspheres were highly effective in loading and prolonged release of both small molecule [dexamethasone (Dex)] and protein [bovine serum albumin (BSA)] models. This strategy has enabled us to control the surface topography of BCP microspheres at defined compositions and holds tremendous promise for drug delivery and tissue engineering applications., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Health status of offspring is programmed by maternal diet throughout gestation and lactation. The present study investigates the lasting effects of maternal supplementation with different amounts of soy oil or extra virgin olive oil (EVOO) on weight and biochemical parameters during gestation and lactation of female mice offspring., Methods: Eight weeks old female C57BL/6 mice (n=40) were assigned through simple randomization into four isocaloric dietary groups (16% of calories as soy oil (LSO) or EVOO (LOO) and 45% of calories as soy oil (HSO) or EVOO (HOO)) during three weeks of gestation and lactation. After weaning (at 3 weeks), all offspring received a diet containing 16% of calories as soy oil and were sacrificed at 6 weeks. Two-way ANOVA was used to adjust for confounding variables and repeated measures test for weight gain trend. Statistical analyses were performed with the IBM SPSS package., Results: At birth and adolescence, the weight of offspring was significantly higher in the soy oil than the olive oil groups (P<0.001 and P<0.001, respectively). Adolescence weight was significantly higher in the offspring born to mothers fed with 16% oil than those with 45% oil (P=0.001). Serum glucose, triglyceride and total cholesterol were significantly higher in the LSO than LOO (P<0.001, P<0.001 and P<0.001), LSO than HSO (P<0.001, P=0.03 and P<0.001), and LOO than HOO (P<0.001, P<0.001 and P<0.001) dietary groups, respectively. Serum triglyceride and total cholesterol were significantly higher in the offspring of HSO than HOO fed mothers (P<0.001 and P<0.001, respectively)., Conclusion: A maternal diet containing EVOO has better effects on birth weight, as well as weight and serum biochemical parameters in offspring at adolescence.

Biological and biomimetic decellularized scaffolds can mimic a natural tissue environment to derive cell proliferation and differentiation without eliciting adverse immune responses during tissue regeneration. Polymeric nanofibrous membranes also served as appropriate matrices for cellular behavior because of their resemblance to physical dimensions of natural extracellular matrix (ECM), while they often have insufficient biological cues to address the cellular phenotype. In this study, we designed bio-engineered membranes through covalent immobilization of decellularized ECM (DECM) particles on the surface of electrospun nanofibers and examined the ability of these composite materials for chondrogenesis. After successful chemical decellularization of human nasal septum cartilage constructs (hNSCs), mechanical processing was carried out and particles with a diameter mean size of 5.06 ± 2.70 μm were yielded. Poly hydroxyalkanoate (PHA) nanofibrous scaffolds were functionalized with DECM particles to mimic the natural motifs of cartilage ECM. Human adipose derived stem cells (hASCs) and human primary chondrocytes (hPChs) cultured on these biofunctional scaffolds showed a significant increase in collagen formation and chondrogenic marker expression after 21 days of cell culture. These results are exciting as they indicate the feasibility of creating bio-engineered scaffolds that may be non-immunogenic as a replacement tissue and have great potential for meeting new challenges in regenerative medicine, particularly in relation to cartilage reconstruction.

Objectives: Maternal high-fat diet has been shown to have deleterious effects on the offspring bones. However, there is no study to assess the effects of type and amount of maternal dietary oil in an isocaloric diet, with focus on extra virgin olive oil (EVOO). The objective of the current study was to test the hypothesis that type of maternal dietary oil has more effects than its amount in an isocaloric diet during gestation and lactation on bone genes expression in offspring in adolescence., Materials and Methods: Virgin female C57BL/6 mice were impregnated and fed either the AIN 93G diet (received 16% of calories as soybean oil, as a control diet, or EVOO) or a high fat AIN 93G diet (received 45% of calories as soybean oil or EVOO) from the time of vaginal plug confirmation until offspring's weaning., Results: After adjusting for the amount of oils, osteoprotegerin/receptor activator of nuclear factor NF-κB ligand (OPG/RANK-L) and OPG expressions were 6.1- and 2.8-folds higher in offspring born to EVOO compared with soybean oil-fed mothers. OPG, beta-catenin, and OPG/RANK-L expression were 88%, 94%, and 70% lower in offspring born to the 45% oil-fed mothers compared with the 16% group. In contrast, peroxisome proliferator-activated receptor gamma-2 (PPARγ2) gene expression was higher in the 45% oil group, adjusted for the types of oil., Conclusion: Maternal EVOO consumption, but not soybean oil increased osteoblastic gene expression, and high amounts of both oils decreased osteoblastic and increased adipogenic genes expression in adolescent offspring.

Bone tissue engineering is sought to apply strategies for bone defects healing without limitations and short-comings of using either bone autografts or allografts and xenografts. The aim of this study was to fabricate a thin layer poly(lactic-co-glycolic) acid (PLGA) coated beta-tricalcium phosphate (β-TCP) scaffold with sustained release of vascular endothelial growth factor (VEGF). PLGA coating increased compressive strength of the β-TCP scaffolds significantly. For in vitro evaluations, canine mesenchymal stem cells (cMSCs) and canine endothelial progenitor cells (cEPCs) were isolated and characterized. Cell proliferation and attachment were demonstrated and the rate of cells proliferation on the VEGF released scaffold was significantly more than compared to the scaffolds with no VEGF loading. A significant increase in expression of COL1 and RUNX2 was indicated in the scaffolds loaded with VEGF and MSCs compared to the other groups. Consequently, PLGA coated β-TCP scaffold with sustained and localized release of VEGF showed favourable results for bone regeneration in vitro, and this scaffold has the potential to use as a drug delivery device in the future., (Copyright © 2016. Published by Elsevier B.V.)

Background: Mesenchymal stem cells (MSCs), have been suggested as a potential choice for treatment of male infertility. Yet, the effects of MSCs on regeneration of germinal epithelium of seminiferous tubules and recovery of spermatogenesis have remained controversial. In this research, we have evaluated and compared the fate of autologous bone marrow (BM)-MSCs during three different periods of time- 4, 6 and 8 weeks after transplantation into the testes of busulfan-induced infertile male rats., Methods: Rats BM samples were collected from tibia bone under anesthesia. The samples were directly cultured in culture medium. Isolated, characterized and purified BM-MSCs were labeled with PKH26, and transplanted into the testes of infertile rats. After 4, 6 and 8 weeks, the testes were removed and underwent histological evaluations., Results: Immunohistochemical analysis showed that transplanted BM-MSCs survived in all three groups. Some of the cells homed at the germinal epithelium and expressed spermatogonia markers ( Dazl and Stella ). The number of homed spermatogonia-like cells in 4-week testes, was more than the 6-week testes. The 8-week testes had the least numbers of homed cells (p<0.05). Immunostaining for vimentin showed that BM-MSCs did not differentiate into the sertoli cells in the testes., Conclusions: From our results, it could be concluded that, autologous BM-MSCs could survive in the testis, migrate onto the seminiferous tubules basement membrane and differentiate into spermatogonia. Although, no more differentiation was observed in the produced spermatogonia, generation of such endogenous GCs would be a really promising achievement for treatment of male infertility using autologous stem cells.

Objective: The aim of this study was to evaluate the effectiveness of the application of cultured autologous bone marrow mesenchymal stem cells (BMSCs) with scaffold and low-level laser therapy (LLLT) on the repair of articular cartilage defects in rabbits., Background Data: For healing of the articular cartilage defects, although positive effects of BMSCs and LLLT have been demonstrated, their combination effect is still unknown; therefore, we investigated combining these two techniques has a synergistic effect., Materials and Methods: After bone marrow aspiration from 10 rabbits, BMSCs were isolated, cultured in monolayer, suspended on a type I collagen scaffold and then implanted onto a full-thickness osteochondral defect (4 mm in diameter), artificially made on the patellar groove of both knees in the same rabbits. Then a knee was selected randomly in each rabbit as the experimental group, and subjected to Ga-Al-As (810 nm) laser irradiation with energy density of 4 J/cm 2 every other day for 3 weeks. As the control group, the other knee did not receive LLLT. After this period, animals were euthanized and osteochondral defects were evaluated by histomorphometric methods., Results: No significant difference in new cartilage formation and inflammation was found between the groups (p > 0.05). However, there was significantly more new bone formation in the experimental group (p < 0.05)., Conclusions: In terms of our research, although better healing in osteochondral defects was seen when combining BMSCs and LLLT compared with the use of BMSCs alone, this improvement was predominantly caused by new bone formation rather than new cartilage formation.

Background: Maternal high fat/high calorie diet leads to adiposity and bone fracture in offspring. However, the effects of macronutrient distribution in maternal isocaloric diet have not been studied. The present study was designed to test the hypothesis that maternal isocaloric pair-fed high-carbohydrate diet will increase osteoblastic and decrease osteoclastic and adipogenic gene expression compared with high-fat diet in adolescent mice offspring., Methods: Virgin female C57BL/6 mice were impregnated and fed either the AIN 93G isocaloric pair-fed high-carbohydrate (LF-HCD) or a high fat (HF-LCD) diet from the time of vaginal plug confirmation until the offspring was weaned., Results: After adjusting for the sex of offspring, osteoprotegrin (OPG) and Ctnnb1 (beta-catenin) genes expression were significantly reduced by 98 % and 97 % in the bone of offspring born from the HF-LCD compared with the LF-HCD-fed mothers ( p  < 0.001 and p  < 0.001, respectively). Peroxisome proliferator-activated receptor gamma-2 (PPAR γ2) gene expression in the bone of offspring born from the HF-LCD was 7.1-folds higher than the LF-HCD-fed mothers ( p  = 0.004). In the retroperitoneal fat mass of offspring born from HF-LCD, AdipoQ and LPL genes expression were respectively up-regulated 15.8 and 4.2-folds compared with the LF-HCD-fed mothers ( p  < 0.001 and p  = 0.03, respectively)., Conclusions: Maternal isocaloric pair-fed high-carbohydrate diet enhances osteoblastogenesis and inhibits adipogenesis compared with high-fat diet in adolescent mice offspring.

This research reports the encapsulation of dexamethasone (Dex) within the chitosan microspheres (CSMs) embedded in a fibrous structure of poly(ɛ-caprolactone) (PCL) to provide a platform for osteogenic differentiation of human mesenchymal stem cells (hMSCs). Dex loaded CSMs were prepared by spray drying a mixture of chitosan and Dex. Then, they were electrospun with PCL solution to create a bilayer fibrous scaffold (PCL/CSMs-Dex). The CSMs act as good depots for sustained release of Dex over a period of 14 days, without noticeable burst release. This is mainly attributed to the core-shell structure of the final PCL/CSMs-Dex-matrix, which could prolong the release and eliminate the initial burst. The water contact angle of PCL scaffolds decreased from 141.4 ± 3.8 to 118.4 ± 7.6 in the presence of CSMs. Improved proliferation of hMSCs cultured on PCL/CSMs-Dex scaffolds was also evidenced. Furthermore, osteogenic assays showed an increase in alkaline phosphatase activity and mineral deposits. The expression of bone-specific genes also confirmed the osteogenic differentiation of cells cultured on these Dex-loaded core-shell structures. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1657-1667, 2016., (© 2016 Wiley Periodicals, Inc.)

Introduction: Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are suitable choices in autologous stem cell treatment of liver-associated diseases due to their hepatic differentiation potential. Dimethyl sulfoxide (DMSO) is an amphipathic molecule with potential of delivering both lipophilic and hydrophilic agents into cells, also a common cryoprotectant for freezing of the cells. DMSO was used in some protocols for induction of AT-MSCs towards hepatocyte like cells. However, the effect of DMSO on hepatogenic differentiation of AT-MSCs were not surveyed, previously. In the present study, we aimed at evaluation of the effect of DMSO on differentiation of AT-MSCs into hepatic lineage., Methods: We isolated mesenchymal stem cells (MSCs) from adipose tissue, and then verifies multi-potency and surface markers of AT-MSCs . Isolated AT-MSCs randomly dispensed in four groups including Group 1: HGF treated, 2: HGF&plus; DMSO treated, 3: HGF&plus; DMSO&plus; OSM treated, and group control for a period of 3 weeks in the expansion medium without serum; EGF and bFGF were also included in the first days of inductions. The morphologic changes during induction period was observed with microscopy. The secretion of albumin (ALB) of the differentiating MSCs was investigated using ELISA, and urea production was evaluated using colorimetric assay. The qRT-PCR was performed for quantitation of hepatocyte marker genes including AFP, ALB, CK18, HNF4a, and HNF6. The glycogen storage of differentiated cells was visualized by periodic-acid Schiff&lsquo;s staining., Results: The results demonstrate that DMSO speeds up hepatic differentiation of AT-MSCs characterized by rapid changes in morphology; higher expression of hepatic marker gene (ALB) in both mRNA and protein level (P &lt; 0.05); also increased transcriptional levels of other liver genes including CK18, HNF4a, and HNF6 (P &lt; 0.01); and moreover, greater percentage of glycogen storage(p &lt; 0.05) in DMSO-treated groups., Conclusion: DMSO catalyzes hepatic differentiation; therefore, using DMSO for acceleration of the hepatogenic protocols of AT-MSCs appears advantageous.

Mesenchymal stem cells (MSCs) have the capacity to differentiate into germ cells (GCs). This research, for the first time, has evaluated the fate of in vitro MSC-derived GCs generated by three different induction methods and compared them after transplantation into the testes of rams. Passage-3 ram bone marrow (BM)-MSCs were divided into three treatment groups: (1) 14-d treatment with 10 μM retinoic acid (RA; RA14), (2) 21-d treatment with 10 μM RA (RA21), and (3) 21-d treatment with 10 ng/ml transforming growth factor beta-1 (TGFb1). After confirmation of the existence of germ-like cells in the culture, the treated cells were labeled and transplanted into the testes of ram lambs. After 2 mo, we conducted histological evaluations of the rams' testes. Results showed that in vitro-derived GCs from all treatment groups survived in the testes. Some of these GCs homed at the basement membrane of seminiferous tubules and formed colonies. The homed cells and cell colonies were similar to testicular native spermatogonia and expressed PGP9.5. TGFb1 exhibited the highest efficiency for in vitro production of GCs as well as the highest capability for homing and colony formation in the testes. RA21 was less efficient than TGFb1, particularly in colony formation. RA14 was the weakest group. No further differentiation of the transplanted GCs was observed. From our results, it could be concluded that a 21-d treatment period of BM-MSCs with TGFb1 is the most efficient method for in vitro generation of spermatogonia-like cells that survive, home, and form colonies in the testes.