Proliferation Of Bone Marrow-derived Stem Cells Research Articles

The effective role of sodium copper chlorophyllin on the dysfunction of bone marrow mesenchymal stem cells in multiple myeloma via regulating TGF-β1

BackgroundThe osteoblast differentiation of bone marrow-derived stem cells (BMSCs) is impaired in multiple myeloma (MM). We investigated the effects of sodium copper chlorophyllin (SCC) on osteoblast differentiation ability of BMSCs from MM. MethodsClinical bone marrow samples were collected. Fluorescence Activated Cell Sorter (FACS) was used to identify surface markers of BMSCs. BMSCs were treated with different concentrations of SCC and cell viability was detected by MTT assay. Relative mRNA and protein expressions of transforming growth factor-β1 (TGF-β1), SMAD2/3, osteogenic differentiation indicators (RUNX2 and OCN) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Alkaline phosphatase (ALP) was stained for activity detection. Formation of calcium nodus of BMSCs was examined by Alizarin Red S staining. ResultsCD90 and CD105 were high-expressed, but CD34 and CD45 were not expressed in BMSCs. BMSCs in MM group showed a lower expression of TGF-β1 and a lower degree of osteogenic differentiation. SCC enhanced activities of BMSCs, ALP activity, and formation of calcium nodus, activated TGF-β1, SMAD2/3 pathway and increased RUNX2 and OCN expressions in BMSCs. Silencing TGF-β1 reversed the effects of SCC on BMSCs in MM. ConclusionSCC could effectively improve the proliferation and osteogenic differentiation of BMSCs in MM through regulating TGF-β1.

Erratum: Intensified Stiffness and Photodynamic Provocation in a Collagen-Based Composite Hydrogel Drive Chondrogenesis.

[This corrects the article DOI: 10.1002/advs.201900099.].

Chitosan/iota-carrageenan and chitosan/pectin polyelectrolyte multilayer scaffolds with antiadhesive and bactericidal properties

It has been a challenge to develop durable and cytocompatible antibacterial coatings with antiadhesive and antimicrobial activities. To overcome the problems caused by bacteria contamination on biomedical devices, we are proposing layer-by-layer films based on iota-carrageenan/chitosan, and pectin/chitosan polyelectrolyte multilayers (PEMs) assembled in an acetic acid/acetate buffer solution at pH 5.0. PEMs with 5 and 15 layers are established with chitosan-terminated layers. As compared to pectin, the iota-carrageenan promotes a PEM with a more wettable surface (water contact angle of 25°) and a low roughness. The PEMs have strong antiadhesive and bactericidal activities against Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus). For P. aeruginosa, the effect is seen after 6 h and for S. aureus the effect is seen after 24 h. The antibacterial test in-vitro indicates that PEMs can kill and avoid the attachment of bacteria effectively. To confirm the biomedical potential of these PEMs to act as coatings for tissue scaffolds and implants, adhesion and proliferation of bone marrow-derived stem cells (BMSCs) is demonstrated after 4 and 7 days of incubation. The iota-carrageenan/chitosan PEM supports the BMSCs adhesion, proliferation and spreading, inhibits the attachment and growth of bacteria and promotes cytocompatibility of surfaces.

Intensified Stiffness and Photodynamic Provocation in a Collagen-Based Composite Hydrogel Drive Chondrogenesis.

Directed differentiation of bone‐marrow‐derived stem cells (BMSCs) toward chondrogenesis has served as a predominant method for cartilage repair but suffers from poor oriented differentiation tendency and low differentiation efficiency. To overcome these two obstacles, an injectable composite hydrogel that consists of collagen hydrogels serving as the scaffold support to accommodate BMSCs and cadmium selenide (CdSe) quantum dots (QDs) is constructed. The introduction of CdSe QDs considerably strengthens the stiffness of the collagen hydrogels via mutual crosslinking using a natural crosslinker (i.e., genipin), which simultaneously triggers photodynamic provocation (PDP) to produce reactive oxygen species (ROS). Experimental results demonstrate that the intensified stiffness and augmented ROS production can synergistically promote the proliferation of BMSCs, induce cartilage‐specific gene expression and increase secretion of glycosaminoglycan. As a result, this approach can facilitate the directed differentiation of BMSCs toward chondrogenesis and accelerate cartilage regeneration in cartilage defect repair, which routes through activation of the TGF‐β/SMAD and mTOR signaling pathways, respectively. Thus, this synergistic strategy based on increased stiffness and PDP‐mediated ROS production provides a general and instructive approach for developing alternative materials applicable for cartilage repair.

An injectable collagen-genipin-carbon dot hydrogel combined with photodynamic therapy to enhance chondrogenesis

Collagen has been widely used for cartilage repair, but its low stiffness and rapid degradation disfavor chondrogenesis. Here we conjugated biocompatible carbon dot nanoparticles (CD NPs) onto collagen through a natural product crosslinker (genipin) to prepare an injectable hydrogel (termed collagen-genipin-CD nanoparticles, CGN). The CGN hydrogel showed increased stiffness due to the cross-linking effect of genipin and the presence of CD NPs, and could produce a moderate amount of reactive oxygen species (ROS) by photodynamic therapy (PDT). Both the stiffness enhancement and ROS generation resulted in improved chondrogenic differentiation of bone marrow-derived stem cells (BMSCs) and the subsequent enhanced cartilage regeneration for cartilage defect repair. Specifically, the CGN hydrogel presented a 21-fold higher compression modulus and a 39.3% lower degradation rate than the pure collagen hydrogel. A combination of both PDT and CGN hydrogel increased the BMSCs proliferation by 50.3%, upregulated their expression of cartilage-specific genes by multiple folds, and enhanced GAG secretion by 205.1% on day 21. This combination also accelerated the cartilage regeneration within as short as 8 weeks. The stiffness enhancement and ROS generation synergistically contributed to chondrogenic differentiation by regulating the TGF-β/SMAD and mTOR signaling pathway, respectively. The combination of CD-modified hydrogel injection and PDT treatment represents a new strategy for minimally invasive repair of cartilage defects.

Aligned Collagen Scaffolds Enhance Tenogenic Differentiation of Bone Marrow-Derived Stem Cells

Tendon repair remains a formidable challenge in clinic, due to its limited function in long-term recovery. Decellularized native tendon extracellular matrix (ECM), which contains specific bioactive factors and a large number of collagen fibers, seeded with bone marrow-derived stem cells (BMSCs) has been found to promote tendon neogenesis. However, the effect of arrangements of natural collagen fibers on the differentiation of BMSCs remains unclear. Here, we prepared aligned collagen fiber scaffolds from rat tail tendon ECM to investigate the effect of collagen arrangements on the morphology, proliferation and differentiation of rat BMSCs in vitro. It was found that BMSCs were elongated and aligned along the collagen fibers on the aligned collagen fibers. Furthermore, the aligned collagen scaffolds significantly enhanced the expression of tenocyte markers TNMD and THBS4. The results indicated that aligned collagen scaffolds could induce the tenogenesis of BMSCs, which would provide an alternative approach for tendon tissue engineering.

Preconditioning with far-infrared irradiation enhances proliferation, cell survival, and migration of rat bone marrow-derived stem cells via CXCR4-ERK pathways

Far-infrared radiation (FIR) has been shown to exert positive effects on the cardiovascular system. However, the biological effects of FIR on bone marrow-derived stem cells (BMSCs) are not understood. In the present study, BMSCs were isolated from rat femur bone marrow and cultured in vitro. To investigate the effects of an FIR generator with an energy flux of 0.13 mW/cm2 on rat BMSCs, survival of BMSCs was measured by crystal violet staining, and cell proliferation was additionally measured using Ez-Cytox cell viability, EdU, and Brd U assays. FIR preconditioning was found to significantly increase BMSC proliferation and survival against H2O2. The scratch and transwell migration assays showed that FIR preconditioning resulted in an increase in BMSC migration. qRT-PCR and Western blot analyses demonstrated that FIR upregulated Nanog, Sox2, c-Kit, Nkx2.5, and CXCR4 at both the mRNA and protein levels. Consistent with these observations, PD98059 (an ERK inhibitor) and AMD3100 (a CXCR4 inhibitor) prevented the activation of CXCR4/ERK and blocked the cell proliferation and migration induced by FIR. Overall, these findings provide the first evidence that FIR confers a real and significant benefit on the preconditioning of BMSCs, and might lead to novel strategies for improving BMSC therapy for cardiac ischemia.

Osteogenesis and chondrogenesis of biomimetic integrated porous PVA/gel/V-n-HA/pa6 scaffolds and BMSCs construct in repair of articular osteochondral defect.

A novel bi-layered osteochondral scaffold, including of PVA/Gel/V layer for the cartilage and n-HA/PA6 layer for the subchondral bone, has been proposed to evaluate the potential of the engineered of osteochondral grafts in repairing articular osteochondral defects in rabbits. The two different layers of the scaffolds were seeded with allogenic bone marrow-derived stem cells (BMSCs), which were chondrogenically and osteogenically induced respectively. The critical-size osteochondral defects were created in the knees of adult rabbits. The defects were treated with cell-bi-layered constructs (Group A), bi-layered constructs (Group B) and untreated group C as control group. The adhesion, proliferation and differentiation of BMSCs were demonstrated by immunohistochemical staining and scanning electron microscopy (SEM) in vitro. Cell survival was tracked via fluorescent labeling in vivo. Overall, the porous PVA/Gel/V-n-HA/PA6 scaffold was compatible and had no negative effects on the BMSCs in vitro culture. The cell-bi-layered scaffolds showed superior repair results as compared to the control group using gross examination and histological assessment. With BMSCs implantation, the two different layers of the composite biomimetic scaffolds provided a suitable environment for cells to form respective tissue. Simultaneously, the RT-PCR results confirmed the expression of specific extracellular matrix (ECM) markers for cartilaginous or osteoid tissue. This investigation showed that the porous PVA/Gel/V-n-HA/PA6 scaffold is a potential matrix for treatment of osteochondral defects, and the method of using chondrogenically and osteogenically differentiated BMSCs as seed cells on each layer might be a promising strategy in repair of articular osteochondral defect due to enhanced chondrogenesis and osteogenesis.

The utilization of decellularized tendon slices to provide an inductive microenvironment for the proliferation and tenogenic differentiation of stem cells

The extracellular matrix (ECM) microenvironment for the stem cell niches, including but not limited to the biochemical composition, matrix topography, and stiffness, is crucial to stem cell proliferation and differentiation. The purpose of this study was to explore the capacity of the decellularized tendon slices (DTSs) to induce stem cell proliferation and tenogenic differentiation. Rat adult stem cells, including tendon-derived stem cells (TDSCs) and bone marrow-derived stem cells (BMSCs), were identified to have universal stem cell characteristics. The DTSs were found to retain the native tendon ECM microenvironment cues, including the inherent surface topography, well-preserved tendon ECM biochemical composition and similar stiffness to native tendon. When the TDSCs and BMSCs were cultured on the DTSs respectively, the LIVE/DEAD assay, alamarBlue® assay, scanning electron microscopy examination and qRT-PCR analysis demonstrated that the DTSs have the capacity to support these stem cells homogeneous distribution, alignment, significant proliferation and tenogenic differentiation. Taken together, the findings of this study indicate that the DTSs can provide a naturally inductive microenvironment for the proliferation and tenogenic differentiation of TDSCs and BMSCs, supporting the use of decellularized tendon ECM as a promising and valuable approach for tendon repair/reconstruction.

Evaluation of Hybrid Porous Biomimetic Nano-Hydroxyapatite/Polyamide 6 and Bone Marrow-Derived Stem Cell Construct in Repair of Calvarial Critical Size Defect

Nano-hydroxyapatite/polyamide 6 (n-HA/PA6) composite scaffolds were prepared by a combined method of phase separation and particle leaching processing technique. The porous matrices were seeded with allogenic bone marrow-derived stem cells (BMSCs) and implanted in the critical-size calvarial defect (8-mm diameter) in rats. The adhesion, proliferation, and differentiation of BMSCs were demonstrated by (3-{4,5-dimethylthiazol-2yl}-2,5-diphenyl-2H-tetrazoliumbromide) assay, immunohistochemical staining, and scanning electron microscopy in vitro. Cell survival was tracked via fluorescent labeling in vivo. The regeneration process was evaluated by several techniques at 4, 8, and 16 weeks after implantation. According to the results, the porous n-HA/PA6 scaffolds are biocompatible and have no negative effects on the BMSCs in vitro. Nano-hydroxyapatite/polyamide 6 composite combined with BMSCs exhibited good biocompatibility and osteoconductivity and enhanced osteogenesis at the initial stage after implantation. The results demonstrated that porous n-HA/PA6 scaffold is a potential matrix for bone tissue engineering, and using BMSCs as seed cells on porous n-HA/PA6 might be an alternative procedure in repair of large defect due to enhanced osteogenesis and osteointegration.

Human bone marrow-derived stem cell proliferation is inhibited by hepatocyte growth factor via increasing the cell cycle inhibitors p53, p21 and p27

Human bone marrow-derived stem cells (hMSCs) are a major source of osteoprogenitors. Hepatocyte growth factor (HGF), a glycoprotein constitutively produced by hMSCs, is reported to act on differentiated osteoblasts and also osteoclasts. Moreover, HGF has been shown by us and others to enhance osteoblastic differentiation from hMSCs. Typically, the pro-differentiation effects of HGF have required cooperative action with regulatory factors such as vitamin D or bone matrix material. Here, we have pursued the molecular mechanisms underlying the osteogenic effect of HGF on hMSCs, the principal precursors to bone forming cells. HGF treatment of hMSCs reduced the cell number over time and increased G1/S cell-cycle arrest compared to control (non-treated) cells. RT-qPCR showed treatment with HGF increased gene expression of the cell-cycle inhibitors p53, p21, and p27, possibly explaining the cell growth inhibition and G1 arrest, a step critical to phenotypic differentiation. Transfection of siRNA specific for cMet, the HGF receptor, eliminated the HGF anti-proliferation effect on hMSCs and the HGF-mediated increase in p53, p21, and p27, strongly supporting a role for these cell-cycle inhibitors in HGF's regulation of hMSCs. HGF in combination with a known inducer of osteogenic differentiation, 1,25-dihydroxyvitamin D, significantly increased cell maturation/differentiation as indicated by an increase in several osteoblast markers. Taken together these results demonstrate that HGF significantly enhances hMSC osteoblast differentiation by 1,25-dihydroxyvitamin D.

Bone marrow‐derived stem cells contribute skin regeneration in skin and soft tissue expansion

Skin and soft tissue expansion stimulates the proliferation of skin epidermal basal cells and increase the dermal collagen deposition and angiogenesis. To explore the contribution of bone marrow-derived stem cells (BMSCs) to the generation of "new" skin during the expansion, we used a chimeric mouse model in which the donor C57BL mice were engrafted with the bone marrow of enhanced green fluorescent protein (EGFP) transgenic mice. BMSCs were collected from the tibia and femur of EGFP(+) transgenic mice, and then injected into normal C57BL mice via the tail vein (chimeric mice). Skin was obtained at different times (days 0, 7, 14, 21, 28, and 35). Skin stromal-derived factor-1 (SDF-1) expression was evaluated. The number, distribution, and phenotype changes of EGFP(+) cells in the skin were also evaluated by means of fluorescent microscopy. EGFP(+) cells were present stably in the normal skin. The number of EGFP(+) cells of the Group A mice changed with the tension, and reached the peak on day 21(17.1 ± 6.7%), as compared with either Group B (5.5 ± 1.0%) or Group C (5.1 ± 0.9%). The SDF-1 expression in the expanded skin was significant increased (≈11-fold, P < 0.01) compared to non-expanded skin on day 21. Immunofluorescence showed EGFP(+) cells were converted into vascular endothelial cells, epidermal cells, and spindle-shaped dermal fibroblasts. Strain can promote the expression of SDF-1 and facilitate the differentiation and proliferation of BMSCs in the expanded skin.