American journal of stem cells最新文献

Objectives: Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease affecting the joint, which is characterized by injury to the articular cartilage, as well as changes in the synovial and subchondral bone. TMJOA has a high incidence rate, without any effective treatment. Despite the therapeutic potential of mesenchymal stem cells (MSCs) in various diseases, their efficacy in treating TMJOA is constrained by the local hypoxic conditions and elevated reactive oxygen species (ROS) environment within the damaged temporomandibular joint. In recent years, many studies have reported that parathyroid hormone (PTH) can effectively treat TMJOA, and has an important impact on MSC differentiation. Therefore, we hypothesized that PTH may influence the potential of MSCs, thereby improving their therapeutic effect on TMJOA.

Methods: First, we isolated and cultured rat bone marrow MSCs, and evaluated their proliferation and differentiation after adding PTH. Next, the in vitro environment of hypoxia and high ROS was established by hypoxia condition and H₂O₂ treatment, and the resistance of PTH-treated MSCs to hypoxia and ROS was subsequently investigated. Finally, PTH-treated MSCs were used to treat TMJOA in a rat model to evaluate the efficacy of PTH.

Results: PTH enhanced the proliferation ability of MSCs, promoted the osteogenic differentiation of MSCs, and improved the tolerance of MSCs to hypoxia and ROS. Finally, the therapeutic effect of PTH-treated MSCs on TMJOA was significantly improved.

Conclusion: PTH enhances the therapeutic effect of MSCs on TMJOA in rats.

Osteonecrosis of the femoral head (ONFH) is a debilitating condition that predominantly affects young individuals, resulting in disability and involving significant healthcare costs. Therefore, it is crucial to develop an effective therapeutic strategy to treat this debilitating disease. In this context, autologous bone marrow-derived mesenchymal stem cells (auto-BM-MSCs) have emerged as a promising approach for treating ONFH. In this case report, we applied this therapy to a patient with ONFH and evaluated both its safety and therapeutic benefits. The treatment consisted of the administration of a single dose of 4×10⁷ ex vivo-expanded auto-BM-MSCs combined with biomolecules derived from platelet-rich plasma. These therapeutic agents were injected into the necrotic zone after accessing it through the technique of multiple small drillings. Subsequently, the progression of ONFH was assessed after 18 months of the auto-BM-MSC administration. Radiographic evaluation showed that the initial femoral head flattening persisted, but no further progression or coxofemoral arthritic changes were observed. Nevertheless, magnetic resonance imaging (MRI) demonstrated a significant improvement in the affected femoral head's area, resulting in a Kerboull angle of 80°, without evidence of flattening or a notable collapse compared to the preoperative condition. Furthermore, the patient exhibited a remarkable functional improvement, as evidenced by a modified Harris hip score of 90 points. The absence of any additional surgery reinforces the positive outcomes achieved through this therapeutic intervention. In conclusion, our case study provides evidence for using the ex vivo-expanded auto-BM-MSCs in combination with platelet-rich plasma-derived biomolecules as a viable and safe treatment for ONFH. However, further research and clinical trials are necessary to validate these promising findings.

Chronic ischemic heart disease remains a major cause of morbidity and mortality worldwide. Several trials have been performed to evaluate benefit of stem cells transplantation to restore cardiac function in short- and long-term period after myocardial infarction. This narrative review analyzes 24 clinical trials between 2005 and 2023 comprising 1824 patients with chronic heart disease without heart failure. Percent increase in left ventricular ejection fraction (LVEF) and decrease in New York Heart Association (NYHA) class at 6/12 months after stem cells transplantation are reported. Thirteen trials showed a statistically significant percent LVEF increase between 4% to 19% at 6/12 months after stem cells transplantation (p values from 0.05 to 0.0001). No significant differences in LVEF were observed between patients who underwent intracoronary or intramyocardial transplantation. NYHA class decrease from severe to mild/moderate was demonstrated in 10 trials reporting a significant LVEF increase. Patients transplanted with bone marrow and peripheral blood CD133+ stem cells showed a doubling of percentage LVEF increase in comparison to patients transplanted with CD133- cells. This narrative review reports the conflicting results on this topic. Multicenter randomized clinical trials should be performed to define the efficacy of stem cells transplantation in chronic ischemic heart disease.

Background: Adipose-derived mesenchymal stem cells (ADSCs) hold promise for bone tissue engineering because of their ability to differentiate into a variety of cell lineages. In tissue engineering, composite scaffolds made of natural and synthetic polymers have also attracted interest. Modification of scaffolds with various substances, including Aloe Vera, is expected to play a useful role in the repair of damaged tissues, including bone.

Method: ADSCs were isolated and seeded in three groups on an Aloe Vera-modified PCL scaffold: 1. Polycaprolactone (PCL) scaffold group, 2. PCL/Aloe Vera scaffold group, and 3. TCPS (Tissue Culture Polystyrene) group. Subsequently, staining with Oil red and Alizarin Red was performed to assess the ability of ADSCs to differentiate into fat and bone cells. Cell viability was determined by the resazurin assay on days 1, 3, and 5. Calcium content and alkaline phosphatase activity (ALP) were determined with kits on days 7, 14, and 21. RNA was extracted, and cDNA was synthesized. Finally, the expression of marker genes for bone differentiation like osteogenic markers such as Osteonectin (ON), Osteocalcin (OC), RUNX Family Transcription Factor 2 (RUNX2), Collagen type I alpha 1 (COL1) was evaluated by real-time PCR.

Results: Aloe vera-treated PCL scaffolds showed improved biocompatibility compared with untreated scaffolds (P<0.05). In addition, treated scaffolds promoted osteogenic differentiation of ADSCs, as evidenced by increased expression of osteogenic markers such ON, OC, RUNX2, COL1 compared with PCL scaffold and TCPS (P<0.05). Furthermore, ALP and calcium content assay confirmed improved mineral deposition on PCL scaffolds treated with Aloe vera, indicating enhanced osteoconductivity (P<0.05).

Conclusion: Our data suggest that a PCL scaffold mixed with Aloe Vera gel has promising osteoconductive potential, which can be used as a natural polymer for tissue engineering of bone and promote bone regeneration.

Background and objectives: Breast cancer stem like cells (CSCs) as a subset of cancer cells exhibit similar properties with normal stem cells. These cells are responsible for cancer metastasis and recurrence. Pivotal roles of CXCR4 in metastasis, chemoresistance and stemness of tumor cells have been showed previously. Here, we aim to explore the relationship between CXCR4 and CSCs in primary and metastatic breast tumor cells.

Methods and results: Primary and highly metastatic breast tumor cells were isolated in our laboratory. Spheroid formation was used to confirm the presence of CSCs and their self-renewal capability. CXCR4 expression was evaluated using real-time polymerase chain reaction in monolayer culture and multicellular spheroids. Our data showed that in all tested cells, CXCR4 expression was significantly increased in CSCs. In parallel, compared with primary tumor cells, downregulation of CXCR4 in metastatic tumor cells was confirmed.

Conclusion: These results provided new insights related to significant alteration of CXCR4 expression in multicellular spheroids. Analysis of molecular properties of spheroids could be used to detect molecular and genetic aspects of CSCs and also created a targeted therapeutic strategy against breast CSCs.

Genetic hearing loss has emerged as a significant public health concern that demands attention. Among the various treatment strategies, gene therapy based on gene editing technology is considered the most promising approach for addressing genetic hearing loss by repairing or eliminating mutated genes. The advent of the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas system has revolutionized gene therapy through its remarkable gene editing capabilities. This system has been extensively employed in mammalian gene editing and is currently being evaluated through clinical trials. Against this backdrop, this review aims to provide an overview of recent advances in utilizing the CRISPR-Cas system to treat genetic hearing loss. Additionally, we delve into the primary challenges and prospects associated with the current application of this system in addressing genetic hearing loss.

Objective: This study investigated if silver nanoparticles (AgNps) could promote the proliferation and osteogenic differentiation of mouse embryonic fibroblasts.

Methods: Mouse embryonic fibroblasts were divided into two groups: Group 1 cells were cultured in DMEM/F12 medium and Group 2 cells were cultured in osteogenic medium. Both groups were then treated with 16, 32, or 100 μM AgNps. Fibroblast proliferation and viability were measured using BrdU and MTT methods at varying time points. Alizarin red staining and alkaline phosphatase (ALP) activity were measured to observe fibroblast differentiation into osteoblasts. Proteomics (cytokine array) was used to detect 111 different cytokines during differentiation.

Results: AgNps stimulated proliferation of mouse embryonic fibroblasts at a concentration of 16 μM. Marked enhancement of calcium mineralization was observed in cells cultured with AgNps compared with cells cultured without AgNps. Group 2 cells displayed nodules around the center where the cell density was high. ALP activity of mouse embryonic fibroblasts cultured in osteogenic medium increased during the whole culture period. Addition of AgNps at concentrations of 32 μM and 100 μM induced higher ALP activity at days 7 and 14. Proteomic array results show that low density lipoprotein receptor (LDL-R) and proprotein convertase subtilisin/kexin type 9 (PCSK-9) were significantly increased, while osteoprotegerin (OPG) was significantly reduced in medium containing 16 μM AgNPs.

Conclusion: AgNps could promote differentiation of mouse embryonic fibroblasts into osteoblastic cells. LDL-R and PCSK-9, as well as OPG, may play a critical role in this process.

Introduction: Autologous adipose-derived stromal vascular fraction (SVF) has been described to have therapeutic benefits in the treatment of keloids. However, most of the evidence on its efficacy is based on observational studies the majority of which are conducted in high-income countries and yet the highest burden of keloids is in low- and middle-income countries (LMICs).

Objectives: We set out to determine the safety and feasibility of using autologous adipose derived stromal vascular fraction in the treatment of keloids in LMICs.

Methods: In this phase II randomized controlled pilot clinical trial conducted in the Plastic Surgery Unit of Kirruddu National Referral Hospital in Kampala Uganda, 8 patients were assigned a 1:1 ratio to either SVF or triamcinolone acetonide (TAC) arms. In the SVF arm, a median (Inter quartile range) amount of stromal cell infiltration of 2.7×10⁶ (11×10⁶) was administered, while the controls received 10 mg/ml TAC at a ratio of 1:1 TAC to keloid volume. Primary endpoints were adverse event development based on the Common Terminology Criteria for Adverse Events (CTCAE) v5.0 tool and feasibility assessment based on ≥ 70% recruitment feasibility and ≥ 80% interventional feasibility rates.

Results: The participants' mean age was 27.9 (±6.5) years, with a female predilection of 5 (63%). Overall, no adverse events were reported in the SVF arm, while ulceration in a single patient in the TAC arm, which was a grade II adverse event, was reported. Recruitment feasibility of 80% and interventional feasibility with 100% completion were reported.

Conclusion: Based on our findings, an autologous adipose-derived stromal vascular fraction is feasible and safe for the treatment of keloids in LMICs.

Background: Basic biological science research deals with nucleic acid isolation. Post-isolation nucleic acid integrity has a pivotal role in further elucidating gene expression and other molecular mechanisms. RNA (ribonucleic acid), cDNA (complementary deoxyribonucleic acid), and PCR (Polymerase chain reaction) products' integrity and quality are affected by several factors in biochemical and biophysical degradation modes. Inadequate evidence was noted about the direct effects of sodium hypochlorite and L-ascorbic acid.

Objectives: This study aims to test the effects of sodium hypochlorite (SHC) and L-ascorbic acid (LAA) in total RNA and PCR products, respectively, in an acellular condition.

Methods: The study was categorized into three steps total RNA, cDNA, and PCR product evaluations. mBM-MSCs were used to extract RNA and then treated with SHC. Crude total RNA and, after DNase 1 treatment, the bands of total RNA samples were visualized by agarose gel electrophoresis. cDNAs were synthesized from SHC-treated (0.25%) and untreated RNAs, which were also expressed on the gel. LAA (5 µM, 15 µM, 25 µM, and 50 µM) were added to cDNAs synthesized from SHC- and non-SHC-treated samples. Housekeeping genes, Gapdh (Glyceraldehyde 3-phosphate dehydrogenase), and 18S rRNA (18S Ribosomal ribonucleic acid) were amplified in both groups.

Results: SHC-treated samples produced clearer bands on an agarose gel. Its treatment did not affect the integrated densities of agarose bands which revealed non-significant (P ≤ 0.05) differences in SHC-treated, untreated RNA, and cDNA. However, significant variations were observed at the PCR level. SHC-treated samples expressed decreased housekeeping gene expression in amplified products (Gapdh and 18S rRNA) and slightly but non-significantly high band intensities appeared in the presence of LAA. Significant variable differences (*P ≤ 0.05) were observed between SHC-treated and non-treated groups after LAA treatment.

Conclusions: SHC (0.25%) is favorable in removing RNases and maintaining the integrity of RNA. cDNA synthesis did not affect by SHC treatment, and it follows the same as untreated samples after DNase 1 treatment. LAA drew a positive impact to improve the quality of PCR products in terms of band intensities, which is insignificant in SHC-treated RNA. Interestingly, it was revealed from our study that 5-25 µM LAA has the most beneficial role in the acquisition of PCR products, i.e. gene expression. These concentrations can be safely used to improve the quality of gene expression. This phenomenon can be used to achieve other, rarer, desired gene expressions. Further research is needed to explore the effects of SHC on the acquisition of PCR products using other solutions.

Objectives: To identify the effect of adipose-derived mesenchymal stem cell-loaded β-chitin nanofiber (ADSC-loaded β-ChNF) hydrogel on diabetic wound healing and clarify its mechanism of action.

Methods: We prepared the ADSC-loaded β-ChNF hydrogel to repair wounds of db/db diabetic mice. Wound healing rate, histopathology, enzyme-linked immunosorbent assay, and western blot were used to confirm its role and mechanism in promoting diabetic wound healing.

Results: The ADSC-loaded β-ChNF hydrogel accelerated wound healing in db/db diabetic mice, as indicated by increased cell proliferation, epithelization, and tissue granulation in the skin. Moreover, expression of vascular endothelial growth factor (VEGF) and its receptor (VEGFR), matrix metalloproteinase 9 (MMP9), and TIMP metallopeptidase inhibitor 1 (TIMP1) were upregulated. These results demonstrate the beneficial effects of this ADSC-loaded β-ChNF hydrogel on diabetic wound healing. Furthermore, we show that the ADSC-loaded β-ChNF hydrogel activated aldolase A (AldoA)/hypoxia-inducible factor 1α (HIF-1α) signaling. An inhibitor of HIF-1α markedly decreased the promotive effects of the ADSC-loaded β-ChNF hydrogel on wound healing and reduced expression of VEGF, VEGFR, MMP9, and TIMP1.

Conclusions: Our findings suggest that the ADSC-loaded β-ChNF hydrogel activated the HIF-1α/MMP9 axis through AldoA feedback to promote diabetic wound healing.