The Efficacy of Transplanting Human Umbilical Cord Mesenchymal Stem Cell Sheets in the Treatment of Myocardial Infarction in Mice.

The transplantation of mesenchymal stem cell (MSC) sheets derived from human umbilical cords (hUCs) was investigated in this study as a potential application in treating myocardial infarction (MI). Two groups of hUC-MSC sheets were formed by populating LunaGelTM, which are 3D scaffolds of photo-crosslinkable gelatin-based hydrogel with two different cell densities. An MI model was created by ligating the left anterior descending coronary artery of healthy BALB/c mice. After two weeks, the cell sheets were applied directly to the MI area and the efficacy of the treatment was evaluated over the next two weeks by monitoring the mice's weight, evaluating the left ventricle ejection fraction, and assessing the histology of the heart tissue at the end of the experiment. Higher cell density showed significantly greater efficiency in MI mice treatment in terms of weight gain and the recovery of ejection fraction. The heart tissue of the groups receiving cell sheets showed human-CD44-positive staining and reduced fibrosis and apoptosis. In conclusion, the hUC-MSC sheets ameliorated heart MI injury in mice and the efficacy of the cell sheets improved as the number of cells increased.

Encapsulation of Human Umbilical Cord Mesenchymal Stem Cells in LunaGel Photocrosslinkable Extracellular Matrix and Subcutaneous Transplantation in Mice.

Stem cells have significant potential in regenerative medicines. However, a major issue with implanting stem cells in the regeneration of new tissue is the methods to implant them and cell viability and functions before and after implantation. Here we developed a simple yet effective method that used photo-crosslinkable gelatin-based hydrogel (LunaGelTM) as a scaffold for the encapsulation, expansion, and eventually, transplantation of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) into mice subcutaneously. We demonstrated the proliferation and maintenance of the original expression of mesenchymal stem cell markers as well as the ability to differentiate into mesoderm-derived cells. The hydrogel was highly stable with no signs of degradation after 20 days in PBS. The hUC-MSCs remained viable after transplantation into mice's subcutaneous pockets and migrated to integrate with the surrounding tissues. We showed a collagen-rich layer surrounding the transplanted cell-laden scaffold indicating the effects of growth factors secreted by the hUC-MSCs. A connective tissue layer was found between the implanted cell-laden scaffold and the collagen layer, and immunohistochemical staining results suggested that this tissue was derived from the MSCs which migrated from within the scaffold. The results, thus, also suggested a protective effect the scaffold has on the encapsulated cells from the antibodies and cytotoxic cells of the host immune system.

Anti-Aging Effects of a Serum Based on Coconut Oil Combined with Deer Antler Stem Cell Extract on a Mouse Model of Skin Aging.

Anti-aging is one of the top goals in the field of health care and aesthetics. Anti-aging cosmetics derived from nature are oriented to long-term development, bringing safety to users and being environmentally friendly. The aim of this study was to develop an anti-aging cosmetic formulation process based on coconut oil in combination with deer antler stem cell extract. The results show that the presence of deer antler stem cell extract added to the foundation made the serum product highly stable and helped improve skin aging significantly after 2 weeks of use. The skin site where the serum product was applied showed a smooth and elastic skin surface, with very few fine lines and shallow wrinkles. Serum reduced the number of wrinkles (48.09% compared to commercial serum (ME) and 60.31% compared to positive control (PC)), reduced skin recovery time (39.31% compared to ME and 67.1% of PC) after two weeks of use. After 2 weeks of use, collagen density increased 10.18% compared to ME and 63.76% compared to control. Epidermal thickness increased by 106.1% compared to PC and 121.7% compared to ME.

A Facile Synthesis Process and Evaluations of α-Calcium Sulfate Hemihydrate for Bone Substitute.

Alpha-calcium sulfate hemihydrate (α-HH) has been used effectively in grafting through its desired features to support bone regeneration. In recent years, many synthetic methods have been proposed. Among them, the autoclave method for manufacturing α-HH is best suited for cost-savings due to its simple operation and limited use of additives. Despite these advantages, the synthesis of surgical grade products without the use of any additives has not yet been clearly discussed. In this study, surgical grade α-HH was successfully produced from calcium sulfate dihydrate (DH) using the autoclave method at an elevated temperature and pressure. The synthesized powder had a high purity of about 98.62% α-HH with a prismatic morphology (20.96 ± 8.83 µm in length and 1.30 ± 0.71 µm in diameter). The screening tests, in simulated body fluid (SBF) solution, for the product properties showed no bioactivity, and fast degradation accompanied by a slight decrease in pH. The lactate dehydrogenase (LDH) assay showed good biocompatibility of the material, however, its potential for cytotoxicity was also observed in NIH 3T3 cells. Briefly, despite some unfavorable properties, the autoclave-synthesized α-HH is a promising bone graft substitute that can be applied in orthopedic and maxillofacial surgeries.

Polydopamine coating of uncrosslinked chitosan as an acellular scaffold for full thickness skin grafts.

There is an unmet need for skin grafting materials that are readily available for large area wounds, due to complex, lengthy and costly manufacturing processes that are not compatible with this type of wounds. Here we developed an acellular skin graft material based on surface coating of uncrosslinked porous (UCLP) chitosan. UCLP chitosan membranes had mechanical properties in ranges suitable for skin grafting. Polydopamine (PDA) coating improved hydrophilicity and resulted in a significant increase in attachment and metabolic activity of mammalian cells in vitro. PDA coating also decreased the attachment of pseudomonas aeruginosa - a common bacteria infecting skin wounds. Finally, the PDA-coated membranes were implanted in full thickness surgical wounds in a rodent model and resulted in complete would closure in 5 days. The current study suggests that PDA-coated UCLP chitosan membranes could be a simple and effective strategy for the development of grafting materials for large area wounds.

Fetal heart extract facilitates the differentiation of human umbilical cord blood-derived mesenchymal stem cells into heart muscle precursor cells.

Human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) are a promising stem cell source with the potential to modulate the immune system as well as the capacity to differentiate into osteoblasts, chondrocytes, and adipocytes. In previous publications, UCB-MSCs have been successfully differentiated into cardiomyocytes. This study aimed to improve the efficacy of differentiation of UCB-MSCs into cardiomyocytes by combining 5-azacytidine (Aza) with mouse fetal heart extract (HE) in the induction medium. UCB-MSCs were isolated from umbilical cord blood according to a published protocol. Murine fetal hearts were used to produce fetal HE using a rapid freeze-thaw procedure. MSCs at the 3rd to 5th passage were differentiated into cardiomyocytes in two kinds of induction medium: complete culture medium plus Aza (Aza group) and complete culture medium plus Aza and fetal HE (Aza + HE group). The results showed that the cells in both kinds of induction medium exhibited the phenotype of cardiomyocytes. At the transcriptional level, the cells expressed a number of cardiac muscle-specific genes such as Nkx2.5, Gata 4, Mef2c, HCN2, hBNP, α-Ca, cTnT, Desmin, and ß-MHC on day 27 in the Aza group and on day 18 in the Aza + HE group. At the translational level, sarcomic α-actin was expressed on day 27 in the Aza group and day 18 in the Aza + HE group. Although they expressed specific genes and proteins of cardiac muscle cells, the induced cells in both groups did not contract and beat spontaneously. These properties are similar to properties of heart muscle precursor cells in vivo. These results demonstrated that the fetal HE facilitates the differentiation process of human UCB-MSCs into heart muscle precursor cells.

Good manufacturing practice-compliant isolation and culture of human umbilical cord blood-derived mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are an attractive source of stem cells for clinical applications. These cells exhibit a multilineage differentiation potential and strong capacity for immune modulation. Thus, MSCs are widely used in cell therapy, tissue engineering, and immunotherapy. Because of important advantages, umbilical cord blood-derived MSCs (UCB-MSCs) have attracted interest for some time. However, the applications of UCB-MSCs are limited by the small number of recoverable UCB-MSCs and fetal bovine serum (FBS)-dependent expansion methods. Hence, this study aimed to establish a xenogenic and allogeneic supplement-free expansion protocol. METHODS: UCB was collected to prepare activated platelet-rich plasma (aPRP) and mononuclear cells (MNCs). aPRP was applied as a supplement in Iscove modified Dulbecco medium (IMDM) together with antibiotics. MNCs were cultured in complete IMDM with four concentrations of aPRP (2, 5, 7, or 10%) or 10% FBS as the control. The efficiency of the protocols was evaluated in terms of the number of adherent cells and their expansion, the percentage of successfully isolated cells in the primary culture, surface marker expression, and in vitro differentiation potential following expansion. RESULTS: The results showed that primary cultures with complete medium containing 10% aPRP exhibited the highest success, whereas expansion in complete medium containing 5% aPRP was suitable. UCB-MSCs isolated using this protocol maintained their immunophenotypes, multilineage differentiation potential, and did not form tumors when injected at a high dose into athymic nude mice. CONCLUSION: This technique provides a method to obtain UCB-MSCs compliant with good manufacturing practices for clinical application.