Proliferation Of Human Mesenchymal Stem Cells Research Articles

Effective production of multifunctional magnetic-sensitive biomaterial by an extrusion-based additive manufacturing technique

A calcium phosphate (CaP)-based scaffold used as synthetic bone grafts, which smartly combines precise dimensions, controlled porosity and therapeutic functions, presents benefits beyond those offered by conventional practices, although its fabrication is still a challenge. The sintering step normally required to improve the strength of the ceramic scaffolds precludes the addition of any biomolecules or functional particles before this stage.This study presents a proof of concept of multifunctional CaP-based scaffolds, fabricated by additive manufacturing from an innovative ink composition, with potential for bone regeneration, cancer treatment by local magnetic hyperthermia and drug delivery platforms. Highly loaded inks comprising iron-doped hydroxyapatite and β-tricalcium phosphate powders suspended in a chitosan-based solution, in the presence of levofloxacin (LEV) as model drug and magnetic nanoparticles (MNP), were developed. The sintering step was removed from the production process, and the integrity of the printed scaffolds was assured by the polymerization capacity of the ink composite, using genipin as a crosslinking agent. The effects of MNP and LEV on the inks’ rheological properties, as well as on the mechanical and structural behaviour of non-doped and iron-doped scaffolds, were evaluated. Magnetic and magneto-thermal response, drug delivery and biological performance, such as cell proliferation in the absence and presence of an applied magnetic field, were also assessed. The addition of a constant amount of MNP in the iron-doped and non-doped CaP-based inks enhances their magnetic response and induction heating, with these effects more pronounced for the iron-doped CaP-based ink. These results suggest a synergistic effect between the iron-doped CaP-based powders and the MNP due to ferro/ferrimagnetic interactions. Furthermore, the iron presence enhances human mesenchymal stem cell metabolic activity and proliferation.

Biomimetic Membranes of Methacrylated Gelatin/Nanohydroxyapatite/Poly(l-Lactic Acid) for Enhanced Bone Regeneration.

Nanofibrous poly(l-lactic acid) (PLLA) membrane-simulated extracellular matrices (ECMs) can be used in the biomedical field. However, the hydrophobic nature and poor osteoinductive property of PLLA limit its application in guided bone regeneration (GBR). In this work, a methacrylated gelatin/nano-HA (GelMA/nHA) complex was first synthesized in situ and then introduced into PLLA to fabricate biomimetic GelMA/nHA/PLLA membranes, mimicking the nanofibrous architecture and composition of ECMs by electrospinning and photocrosslinking. Compared to PLLA and GelMA/PLLA membranes, the novel GelMA/nHA/PLLA membranes demonstrated better tensile, hydrophilic, water sorption, and degradation properties. An in vitro biological evaluation indicated that the membranes promoted human bone marrow-derived mesenchymal stem cell (hBMSC) proliferation, adhesion, and osteogenic differentiation. Critical-sized defects in rat models were used to evaluate the bone regeneration performances of the three kinds of membranes in vivo, and the GelMA/nHA/PLLA membranes demonstrated excellent osteogenic regeneration potential. Therefore, GelMA/nHA/PLLA membranes have wide application prospects in bioengineering applications such as GBR treatment.

Designing topographically textured microparticles for induction and modulation of osteogenesis in mesenchymal stem cell engineering

Mesenchymal stem cells are the focus of intense research in bone development and regeneration. The potential of microparticles as modulating moieties of osteogenic response by utilizing their architectural features is demonstrated herein. Topographically textured microparticles of varying microscale features are produced by exploiting phase-separation of a readily soluble sacrificial component from polylactic acid. The influence of varying topographical features on primary human mesenchymal stem cell attachment, proliferation and markers of osteogenesis is investigated. In the absence of osteoinductive supplements, cells cultured on textured microparticles exhibit notably increased expression of osteogenic markers relative to conventional smooth microparticles. They also exhibit varying morphological, attachment and proliferation responses. Significantly altered gene expression and metabolic profiles are observed, with varying histological characteristics in vivo. This study highlights how tailoring topographical design offers cell-instructive 3D microenvironments which allow manipulation of stem cell fate by eliciting the desired downstream response without use of exogenous osteoinductive factors.

In Situ Generation of Hydroxyapatite on Biopolymer Particles for Fabrication of Bone Scaffolds Owning Bioactivity

Hydroxyapatite (HAP) can endow a biopolymer scaffold with good bioactivity and osteoconductive ability, while the interfacial bonding is fairly weak between HAP and biopolymers. In this study, HAP was in situ generated on poly(l-lactic acid) (PLLA) particles, and then they were used to fabricate a scaffold by selective laser sintering. Detailedly, PLLA particles were first functionalized by dopamine oxide polymerization, which introduced abundance active catechol groups on the particle surface, and subsequently, the catechol groups concentrated Ca2+ ions by chelation in a simulated body fluid solution, and then, Ca2+ ions absorbed PO43- ions through electrostatic interactions for in situ nucleation of HAP. The results indicated that HAP was homogeneously generated on the PLLA particle surface, and HAP and PLLA exhibited good interfacial bonding in the HAP/PLLA scaffolds. Meanwhile, the scaffolds displayed excellent bioactivity by inducing apatite precipitation and provided a good environment for human bone mesenchymal stem cell attachment, proliferation, and osteogenic differentiation. More importantly, the ingrowth of blood vessel and the formation of new bone could be stimulated by the scaffolds in vivo, and the bone volume fraction and bone mineral density increased by 44.44 and 41.73% compared with the pure PLLA scaffolds, respectively. Serum biochemical indexes fell within the normal range, which indicated that there was no harmful effect on the normal functioning of the body after implanting the scaffold.

In-Situ Synthesis and Characterization of Chitosan/Hydroxyapatite Nanocomposite Coatings to Improve the Bioactive Properties of Ti6Al4V Substrates.

Ti6Al4V alloy is still attracting great interest because of its application as an implant material for hard tissue repair. This research aims to produce and investigate in-situ chitosan/hydroxyapatite (CS/HA) nanocomposite coatings based on different amounts of HA (10, 50 and 60 wt.%) on alkali-treated Ti6Al4V substrate through the sol-gel process to enhance in vitro bioactivity. The influence of different contents of HA on the morphology, contact angle, roughness, adhesion strength, and in vitro bioactivity of the CS/HA coatings was studied. Results confirmed that, with increasing the HA content, the surface morphology of crack-free CS/HA coatings changed for nucleation modification and HA nanocrystals growth, and consequently, the surface roughness of the coatings increased. Furthermore, the bioactivity of the CS/HA nanocomposite coatings enhanced bone-like apatite layer formation on the material surface with increasing HA content. Moreover, CS/HA nanocomposite coatings were biocompatible and, in particular, CS/10 wt.% HA composition significantly promoted human mesenchymal stem cells (hMSCs) proliferation. In particular, these results demonstrate that the treatment strategy used during the bioprocess was able to improve in vitro properties enough to meet the clinical performance. Indeed, it is predicted that the dense and crack-free CS/HA nanocomposite coatings suggest good potential application as dental implants.

Competition of carrier bioresorption and drug release kinetics of vancomycin-loaded silicate macroporous microspheres to determine cell biocompatibility

Bioceramic porous microspheres are promising substances for dental and orthopedic bone void filling, tissue engineering and drug delivery applications. In this research, the structure and cytocompatibility of bioactive magnesium-calcium silicate macroporous microspheres loaded with vancomycin hydrochloride, an antibiotic drug, were studied. In this regard, bredigite (Ca7MgSi4O16), akermanite (Ca2MgSi2O7) and diopside (CaMgSi2O6) carriers were fabricated through a sequence of sol-gel, calcination, droplet extrusion and sintering processes, followed by impregnated with vancomycin. X-ray diffraction (XRD) and scanning electron microscopy verified the formation of the desired ceramic crystalline phases and macroporous characteristics of the carriers, respectively. Based on the MTT assay, the antibiotic-loaded bredigite, akermanite and diopside devices comparatively exhibited the lowest, intermediate and highest levels of human bone marrow mesenchymal stem cells (hBM-MSCs) viability and proliferation, respectively. It was concluded that the role of the carrier bioresorption kinetics prevails over the drug delivery kinetics in determining the cell biocompatibility of the devices.

Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation.

AimsHere we introduce a wide and complex study comparing effects of growth factors used alone and in combinations on human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation. Certain ways of cell behaviour can be triggered by specific peptides – growth factors, influencing cell fate through surface cellular receptors.MethodsIn our study transforming growth factor β (TGF-β), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1), and vascular endothelial growth factor (VEGF) were used in order to induce osteogenesis and proliferation of hMSCs from bone marrow. These cells are naturally able to differentiate into various mesodermal cell lines. Effect of each factor itself is pretty well known. We designed experimental groups where two and more growth factors were combined. We supposed cumulative effect would appear when more growth factors with the same effect were combined. The cellular metabolism was evaluated using MTS assay and double-stranded DNA (dsDNA) amount using PicoGreen assay. Alkaline phosphatase (ALP) activity, as early osteogenesis marker, was observed. Phase contrast microscopy was used for cell morphology evaluation.ResultsTGF-β and bFGF were shown to significantly enhance cell proliferation. VEGF and IGF-1 supported ALP activity. Light microscopy showed initial extracellular matrix mineralization after VEGF/IGF-1 supply.ConclusionA combination of more than two growth factors did not support the cellular metabolism level and ALP activity even though the growth factor itself had a positive effect. This is probably caused by interplay of various messengers shared by more growth factor signalling cascades.Cite this article: Bone Joint Res 2020;9(7):412–420.

UHMWPE-MWCNT-nHA based hybrid trilayer nanobiocomposite: Processing approach, physical properties, stem/bone cell functionality, and blood compatibility.

The development of polymeric nanocomposites for biomedical applications remains a major challenge in terms of tailored addition of nanoparticles to realize the simultaneous enhancement of fracture resistance and cell/blood compatibility. To address this, the present work has been planned to determine whether small addition of surface functionalized multiwalled-carbon-nanotube, MWCNT (<1.5 wt%) and egg-shell derived nanosized hydroxyapatite, nHA (<10 wt%) to ultrahigh-molecular-weight-polyethylene (UHMWPE) can significantly improve the physical properties as well as biocompatibility. The difference in mouse osteoblast and human mesenchymal stem cell (hMSc) proliferation has been validated using both the monolithic composite and a trilayered composite with two different UHMWPE nanocomposites on either face with pure polymer at the middle. The combination of rheology and micro-CT with fractography reveals the homogeneous dispersion of nanofillers, leading to mechanical property enhancement. The quantitative analysis of cell viability and cell spreading by immunocytochemistry method, using vinculin and vimentin expression, establish significant cytocompatibility with hMSc and osteoblast cells onto the trilayer hybrid nanobiocomposite substrates. The hemocompatibility of the investigated composites under the controlled flow of rabbit blood in a microfluidic device reveals the signature of reduced thrombogenesis with reduction of platelet activation on UHMWPE nanocomposite w.r.t. unreinforced UHMWPE. An attempt has been made to discuss the blood compatibility results in the backdrop of the bovine serum albumin adsorption kinetics. Summarizing, the present study establishes that the twin requirement of mechanical property and cyto/hemo-compatibility can be potentially realized in developing trilayer composites in UHMWPE-nHA-MWCNT system.

Aminated 3D Printed Polystyrene Maintains Stem Cell Proliferation and Osteogenic Differentiation.

As 3D printing becomes more common and the technique is used to build culture platforms, it is imperative to develop surface treatments for specific responses. The advantages of aminating and oxidizing polystyrene (PS) for human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation are investigated. We find that ammonia (NH3) plasma incorporates amines while oxygen plasma adds carbonyl and carboxylate groups. Across 2D, 3D, and 3D dynamic culture, we find that the NH3- treated surfaces encouraged cell proliferation. Our results show that the NH3-treated scaffold was the only treatment allowing dynamic proliferation of hMSCs with little evidence of osteogenic differentiation. With osteogenic media, particularly in 3D culture, we find the NH3 treatment encouraged greater and earlier expression of RUNX2 and ALP. The NH3-treated PS scaffolds support hMSC proliferation without spontaneous osteogenic differentiation in static and dynamic culture. This work provides an opportunity for further investigations into shear profiling and coculture within the developed culture system toward developing a bone marrow niche model.

Mussel inspired ZIF8 microcarriers: a new approach for large-scale production of stem cells.

Metal–organic frameworks (MOFs) have high porosity, large surface area, and tunable functionality and have been widely used for drug loading. The aim of this study was to exploit unique features of zeolitic imidazolate framework-8 (ZIF8) to develop an innovative composite microcarrier (MC) for human mesenchymal stem cells (hMSCs) adhesion and proliferation. ZIF8 MCs were prepared by immobilization of polydopamine/polyethyleneimine (PDA/PEI) and ZIF8 on the surface of polystyrene beads. The chemical properties of MCs such as coating stability and homogeneity were characterized by different techniques such as ATR-FTIR, XRD, EDX, SEM, and contact angle. The prepared MCs were tested using human adipose-derived mesenchymal stem cells (hADSCs) in both static and dynamic conditions, and results were compared to a commercially available MC (Star-Plus), polydopamine coated MCs and amine-functionalized MCs as a control. Results show that PDA/PEI/ZIF8 coated MCs (in short: ZIF8 MCs) provides an excellent biocompatible environment for hADSCs adhesion and growth. In conclusion, ZIF8 MCs represent suitable and low-cost support for hADSCs culture and expansion, which can maximize cell yield and viability while preserving hADSCs multipotency. The present findings have revealed this strategy has the potential for chemical and topographical modification of MCs in tissue engineering applications.

Strontium-substituted biphasic calcium phosphate microspheres promoted degradation performance and enhanced bone regeneration.

Biphasic calcium phosphate (BCP) ceramics are the subject of much attention for use as bone replacement material. However, it remains a challenge to promote the degradation and osteoinductivity performances of BCP ceramics. In this work, novel BCP ceramic microspheres with good degradation and excellent osteoinductivity were prepared through high-content strontium (Sr) doping. The in vitro results indicated that the Sr10-BCP, Sr40-BCP, and Sr80-BCP microspheres all had their HA crystals partially transformed to the beta tricalcium phosphate phase following high-temperature sintering because of Ca-deficient HA formed by the partial substitution of Ca ions by Sr ions. In addition, the degradation rate was increased with the doping of increasing amounts of Sr. All prepared microspheres enhanced human bone marrow-derived mesenchymal stem cells attachment and proliferation. Specifically, among these modified microspheres, the Sr40-BCP microspheres showed the highest osteogenic potential. Furthermore, Sr40-BCP and HA microspheres were implanted in a calvarial defect model of rat to evaluate the in vivo bone augmentation ability. The results indicated that Sr40-BCP microspheres degraded more completely and significantly promoted new bone regeneration compared with HA microspheres. In conclusion, Sr40-BCP microspheres have excellent potential for degradation and bone regeneration and are promising osteogenic materials.

Adhesion, proliferation and differentiation of human mesenchymal stem cell on chitosan/collagen composite scaffold.

In vitro tissue engineering requires a progenitor cell source and a porous scaffold providing three dimensional (3D) supports for growth and differentiation to attain tissue architectures. This research focused on fabrication and characterization of 3D porous scaffolds using chitosan (CS), collagen (CG) and chitosan-collagen (CS-CG) composite to investigate their influence on human mesenchymal stem cell (hMSC) adhesion, proliferation and differentiation. Material dependent variations in porous morphology and mechanical behavior of the fabricated CS, CG and CS-CG scaffold showed significant impact on hMSC adhesion, proliferation and differentiation. The maximum hMSC adhesion and proliferation was reported on CS-CG scaffold among all fabricated scaffold groups. Interconnectivity of pores structure in CS-CG scaffold was considered as preferable attribute for such enhanced growth and distribution throughout the scaffold. Besides, CS scaffold with well interconnected pores showed poor adhesion and proliferation because of inadequate adhesion motifs. In case of CG scaffold, optimum growth and distribution of hMSC occurs only at the surface because of the absence of interconnectivity in their pore structures. Likewise, osteogenic differentiation of hMSC occurs most preferably in CS-CG composite scaffold among all scaffold groups. Such enhanced hMSC proliferation and differentiation in CS-CG scaffold significantly influenced on mechanical behavior of scaffold which is essential for in vivo application of a bone tissue implant. Thus CS-CG composite scaffold holds promise to be a suitable platform for in vitro engineering of bone tissue implant.

3D printing of layered mesoporous bioactive glass/sodium alginate-sodium alginate scaffolds with controllable dual-drug release behaviors

Scaffolds with controlled drug release are valuable for bone tissue engineering, but constructing the scaffolds with controllable dual-drug release behaviors is still a challenge. In this study, layered mesoporous bioactive glass/sodium alginate-sodium alginate (MBG/SA–SA) scaffolds with controllable dual-drug release behaviors were fabricated by 3D printing. The porosity and compressive strength of three-dimensional (3D) printed MBG/SA–SA scaffolds by cross-linking are about 78% and 4.2 MPa, respectively. As two model drugs, bovine serum albumin (BSA) and ibuprofen (IBU) were separately loaded in SA layer and MBG/SA layer, resulting in a relatively fast release of BSA and a sustained release of IBU. Furthermore, layered MBG/SA–SA scaffolds were able to stimulate human bone mesenchymal stem cells (hBMSCs) adhesion, proliferation and osteogenic differentiation than SA scaffolds. Hence, the 3D printed MBG/SA–SA scaffolds would be prospective for the treatment of bone defects.

Preparation and characterization of ß-lactoglobulin/poly(ethylene oxide) magnetic nanofibers for biomedical applications

Developing a single nanofiber containing magnetic nanoparticles (MGNPs) and whey proteins provides a preferable requirement to utilize the benefits of various amino acids and magnetization characteristics in a single nanostructure. In this study, ß-lactoglobulin (BLG), one of the major components of the aqueous whey protein isolate, and magnetic glass ceramic nanoparticles (MGNPs) colloid were formulated into nanofibrous morphology by electrospinning using the assist of a poly(ethylene oxide) (PEO). First, BLG:PEO with a ratio of 3:1 produced nanofibers with an average fiber diameter of 370 nm. To produce magnetized nanofiber scaffolds, the magnetic glass ceramic nanoparticles were added to the BLG-PEO solution with a ratio of 1, 3 and 7 wt%. The influence of the inorganic nanoparticles on the nanofibrous morphology of the electrospun scaffold was investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The nanofiber diameter decreased with increasing the nanoparticles concentration as the observed average diameters were 245, 163 and 152 nm for the scaffolds containing 1, 3 and 7 wt.% MGNPs, respectively. The chemical composition and the magnetic properties were characterized using FTIR, DSC and VSM analysis techniques. It was concluded that addition of MGNPs did not affect the chemical composition of the protein or the polymer. Accordingly, a cell proliferation assay confirmed that the magnetic nanofibers have no adverse effect on the human mesenchymal stem cell (hMSC) proliferation. These results confirm the possibility of using electrospun magnetic nanofibers for biomedical applications.

3D gelatin-chitosan hybrid hydrogels combined with human platelet lysate highly support human mesenchymal stem cell proliferation and osteogenic differentiation

Bone marrow and adipose tissue human mesenchymal stem cells were seeded in highly performing 3D gelatin–chitosan hybrid hydrogels of varying chitosan content in the presence of human platelet lysate and evaluated for their proliferation and osteogenic differentiation. Both bone marrow and adipose tissue human mesenchymal stem cells in gelatin–chitosan hybrid hydrogel 1 (chitosan content 8.1%) or gelatin–chitosan hybrid hydrogel 2 (chitosan 14.9%) showed high levels of viability (80%–90%), and their proliferation and osteogenic differentiation was significantly higher with human platelet lysate compared to fetal bovine serum, particularly in gelatin–chitosan hybrid hydrogel 1. Mineralization was detected early, after 21 days of culture, when human platelet lysate was used in the presence of osteogenic stimuli. Proteomic characterization of human platelet lysate highlighted 59 proteins mainly involved in functions related to cell adhesion, cellular repairing mechanisms, and regulation of cell differentiation. In conclusion, the combination of our gelatin–chitosan hybrid hydrogels with hPL represents a promising strategy for bone regenerative medicine using human mesenchymal stem cells.

Fabrication and characterisation of 3D printed MWCNT composite porous scaffolds for bone regeneration

Carbon nanotubes (CNTs) with exceptional physical and chemical properties are attracting significant interest in the field of tissue engineering. Several reports investigated CNTs biocompatibility and their impact in terms of cell attachment, proliferation and differentiation mainly using polymer/CNTs membranes. However, these 2D membranes are not able to emulate the complex in vivo environment. In this paper, additive manufacturing (3D printing) is used to create composite 3D porous scaffolds containing different loadings of multi-walled carbon nanotubes (MWCNT) (0.25, 0.75 and 3 wt%) for bone tissue regeneration. Pre-processed and processed materials were extensively characterised in terms of printability, morphological and topographic characteristics and thermal, mechanical and biological properties. Scaffolds with pore sizes ranging between 366 μm and 397 μm were successfully produced and able to sustain early-stage human adipose-derived mesenchymal stem cells attachment and proliferation. Results show that MWCNTs enhances protein adsorption, mechanical and biological properties. Composite scaffolds, particularly the 3 wt% loading of MWCNTs, seem to be good candidates for bone tissue regeneration.

Titanium dioxide nanotubes of defined diameter enhance mesenchymal stem cell proliferation via JNK- and ERK-dependent up-regulation of fibroblast growth factor-2 by T lymphocytes.

Long-term clinical success of a titanium implant not only depends upon osseointegration between implant and bone surface but also on the response of host immune cells. Following implantation of biomaterials, an inflammatory response, including T lymphocyte response, is ostensibly initiated by implant-cell interaction. However, little is known about the responses of T lymphocytes to titanium dioxide nanotubes. The present study aimed to explore the effect of titanium dioxide nanotubes on T lymphocytes in vitro and its biological consequences. The results of the present study showed that titanium dioxide nanotubes with diameter of 30-105 nm were non-cytotoxic to T lymphocytes, and the 105 nm titanium dioxide nanotube surface specifically possessed an ability to activate T lymphocytes, thus increasing DNA synthesis and cell proliferation. In addition, the 105 nm titanium dioxide nanotubes significantly activated the expression of FGF-2 gene and protein in T lymphocytes although smaller nanotubes (i.e. those with diameters of approximately 30 and 70 nm) had little effect on this. The present study investigated the mechanism by which 105 nm nanotubes stimulated FGF-2 expression in T lymphocytes by blocking key MAPK pathways. The inhibitors of JNK1/2/3 and ERK1/2 significantly inhibited 105 nm titanium dioxide nanotubes-induced FGF-2 expression. Corresponding to the increased expression of FGF-2, only the supernatant from T lymphocytes cultured on 105 nm nanotubes stimulated human mesenchymal stem cell proliferation. FGF-2 blocking antibody partially reversed the increased proliferation of human mesenchymal stem cells, supporting the role of T lymphocyte-derived FGF-2 in enhanced human mesenchymal stem cell proliferation. This suggests a significant role of T lymphocyte-titanium dioxide nanotube interaction in the proliferation of human mesenchymal stem cells, which is pivotal to the formation of new bone following implant placement.

3D-printed scaffolds of biomineralized hydroxyapatite nanocomposite on silk fibroin for improving bone regeneration

In an attempt to fabricate biomimetic bone repair scaffolds and improve bone regeneration point of view, we have three dimensionally printed porous scaffolds with biomineralized hydroxyapatite/silk fibroin nanocomposites. SF/HA composite particles were firstly produced via an in-situ mineral precipitation process when SF molecules were served as templates.. Microscopy observations of SF/HA showed homogeneous morphology and narrowly distributed size. By using sodium alginate (SA) as paste binder, scaffolds with different contents of SF/HA were subsequently 3D-printed under proper conditions. All the scaffolds were porous with 3D interconnected large pores (size ～400 μm) and an overall porosity about 70%, combined with a relative high compressive strength (>6 MPa). The in vitro biological properties of degradation, apatite formation, human bone marrow-derived mesenchymal stem cells (hBMSCs) proliferation and differentiation were characterized on SF/HA-SA scaffolds. Results indicated that more apatite newly formed on the scaffold surface after soaking in SBF and original deposited HA minerals would recrystallize. In addition, the pH values of medium microenvironment remained steady in the range from 6.9 to 7.1 in one month. Cell attachment and penetration into scaffolds were supported by all the groups. Increased content of SF/HA led to better cell proliferation, and enhanced ALP activity as well. Bovine serum albumin (BSA) protein was selected as a model drug to be loaded into SF/HA-SA scaffolds in printing process, and the drugs released in a relative sustained manner in 5 days. These results suggest a promising application of SF/HA-SA scaffolds in bone regeneration.

Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

Background3D cell culture is an appropriate method to develop engineered bone tissue, where different bioreactors have been designed to mitigate the challenges in 3D culture. Currently, we tailored a perfusion reactor to witness human mesenchymal stem cells (MSCs) proliferation and differentiation over polylactic acid-polyethylene glycol (PLA/PEG) composite scaffolds. MethodsThe composite scaffolds with different weight ratios of PLA and PEG were prepared using solvent casting-particulate leaching technique. Human umbilcal card blood MSCs were cultured under dynamic and static conditions to elucidate the role of dynamic fluid flow in osteogenesis of MSCs. ResultsThe human MSCs distribution over the scaffolds was confirmed with fluorescent microscopy. Alkaline phosphatase (ALP), calcium mineralization, and collagen formation were found to be higher in PLA90 scaffolds than PLA100 and PLA75. PLA90 scaffolds with better cell adhesion/proliferartion were considered for bioreactor studies and they exhibited enhanced ALP, Ca+2 mineralization and collagen formation under dynamic perfusion than static culture. We further confirmed our observation by looking at expression levels of osteogenic marker (Runx2 and osteonectin) in differentiated MSCs subjected to perfusion culture compared to static culture. ConclusionThe results of the current investigation once again proves that dynamic perfusion cultures improve the osteogenic differentiation of MSCs over hybrid polymer scaffolds (PLA90) for effective bone regeneration.

The C-terminus of the amelogenin peptide influences the proliferation of human bone marrow mesenchymal stem cells.

Amelogenins are a family of enamel matrix proteins that are important for formation of enamel. Amelogenins may induce division of mesenchymal stem cells (MSCs), among others. Recently, the C-terminus of the amelogenin peptide (AMG-CP) has been shown to enhance the proliferation of cementoblast lineage cells. The role of the amelogenin peptide on the proliferation of human MSCs and related alterations in the intracellular signaling pathway were studied. MSCs were exposed to AMG-CP in vitro. The MTS and 5-bromo-2'-deoxyuridine (BrdU) assays were used to determine proliferation. Expression of the amelogenin receptor, lysosomal-associated membrane protein 1 (LAMP1), was examined in MSCs with western blotting. Binding of AMG-CP to LAMP1 was inhibited with anti-LAMP1 antibody. Components of the mitogen-activated protein kinase (MAPK)-extracellular signal-regulated kinase (ERK) pathway were studied with western blotting and enzyme-linked immunosorbent assay, and U0126, an MAPK inhibitor, was used to inhibit ERK activity. MSC proliferation was significantly increased in the presence of AMG-CP and significantly inhibited by anti-LAMP1 antibody or U0126. Increased phosphorylated ERK1/2 was observed in the presence of AMG-CP, and decreased phosphorylated ERK1/2 was seen in the presence of anti-LAMP1 antibody or U0126. A C-terminal amelogenin variant increased the proliferation of MSCs via an interaction with LAMP1 and the MAPK-ERK signaling pathway, indicating the possibility of using MSCs for tissue regeneration in the craniofacial region.