Thursday, December 24, 2009

12/25 TE-RegenMed-StemCell feed

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TE-RegenMed-StemCell feed - By; Ankur Gupta Feed My Inbox

Stanford scientists identify protein that keeps stem cells poised for action
December 24, 2009 at 1:24 pm


Citrus surprise: Vitamin C boosts the reprogramming of adult cells into stem cells
December 24, 2009 at 1:24 pm


Science and Research in Academic Plastic Surgery in Germany.
December 24, 2009 at 6:36 am

Science and Research in Academic Plastic Surgery in Germany.

Handchir Mikrochir Plast Chir. 2009 Dec;41(6):359-363

Authors: Giunta RE, Machens HG

Plastic surgery has passed through a very positive evolution in the last decades on the solid fundament of constantly developing academic plastic surgery. Aim of this paper is an objective evaluation of the current status of academic plastic surgery regarding research topics, currently available ressources and scientific outcome based on a questionnaire. The return rate of the questionnaire in academic departments was 92%. Main topics in research besides wound healing were topics from regenerative medicine such as tissue engineering, biomaterials, genetherapy and angiogenesis with the main focus on skin and fat tissues. In the past five years a total of 25 million Euros of third party research grants were raised. Research relied mainly on interdisciplinary research facilities. Regarding the scientific outcome more than 200 scientific papers were published in basic science research journals having an impactfactor higher than two. These results clearly demonstrate that plastic surgery is scientifically highly productive in academic surroundings where independent departments are established. Considering that independent units of plastic surgery exist in a relatively small number of all 36 university hospitals in germany, it has to be claimed for further independent departments so to provide adequate research facilities for further evolution of academic plastic surgery.

PMID: 20029742 [PubMed - as supplied by publisher]


On the Independent Origins of Complex Brains and Neurons.
December 24, 2009 at 6:36 am

On the Independent Origins of Complex Brains and Neurons.

Brain Behav Evol. 2009;74(3):177-190

Authors: Moroz LL

Analysis of the origin and evolution of neurons is crucial for revealing principles of organization of neural circuits with unexpected implications for genomic sciences, biomedical applications and regenerative medicine. This article presents an overview of some controversial ideas about the origin and evolution of neurons and nervous systems, focusing on the independent origin of complex brains and possible independent origins of neurons. First, earlier hypotheses related to the origin of neurons are summarized. Second, the diversity of nervous systems and convergent evolution of complex brains in relation to current views about animal phylogeny is discussed. Third, the lineages of molluscs and basal metazoans are used as illustrated examples of multiple origins of complex brains and neurons. Finally, a hypothesis about the independent origin of complex brains, centralized nervous systems and neurons is outlined. Injury-associated mechanisms leading to secretion of signal peptides (and related molecules) can be considered as evolutionary predecessors of inter-neuronal signaling and the major factors in the appearance of neurons in the first place.

PMID: 20029182 [PubMed - as supplied by publisher]


Receptor Channel TRPC6 Is a Key Mediator of Notch-Driven Glioblastoma Growth and Invasiveness.
December 24, 2009 at 6:36 am

Receptor Channel TRPC6 Is a Key Mediator of Notch-Driven Glioblastoma Growth and Invasiveness.

Cancer Res. 2009 Dec 22;

Authors: Chigurupati S, Venkataraman R, Barrera D, Naganathan A, Madan M, Paul L, Pattisapu JV, Kyriazis GA, Sugaya K, Bushnev S, Lathia JD, Rich JN, Chan SL

Glioblastoma multiforme (GBM) is the most frequent and incurable type of brain tumor of adults. Hypoxia has been shown to direct GBM toward a more aggressive and malignant state. Here we show that hypoxia increases Notch1 activation, which in turn induces the expression of transient receptor potential 6 (TRPC6) in primary samples and cell lines derived from GBM. TRPC6 is required for the development of the aggressive phenotype because knockdown of TRPC6 expression inhibits glioma growth, invasion, and angiogenesis. Functionally, TRPC6 causes a sustained elevation of intracellular calcium that is coupled to the activation of the calcineurin-nuclear factor of activated T-cell (NFAT) pathway. Pharmacologic inhibition of the calcineurin-NFAT pathway substantially reduces the development of the malignant GBM phenotypes under hypoxia. Clinically, expression of TRPC6 was elevated in GBM specimens in comparison with normal tissues. Collectively, our studies indicate that TRPC6 is a key mediator of tumor growth of GBM in vitro and in vivo and that TRPC6 may be a promising therapeutic target in the treatment of human GBM. Cancer Res; 70(1); 418-27.

PMID: 20028870 [PubMed - as supplied by publisher]


California stem-cell grants awarded.
December 24, 2009 at 6:36 am

Related Articles

California stem-cell grants awarded.

Nature. 2009 Nov 5;462(7269):22

Authors: Check Hayden E

PMID: 19890299 [PubMed - indexed for MEDLINE]


Neurotrophic factor-expressing mesenchymal stem cells survive transplantation into the contused spinal cord without differentiating into neural cells.
December 24, 2009 at 6:36 am

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Neurotrophic factor-expressing mesenchymal stem cells survive transplantation into the contused spinal cord without differentiating into neural cells.

Tissue Eng Part A. 2009 Oct;15(10):3049-59

Authors: Rooney GE, McMahon SS, Ritter T, Garcia Y, Moran C, Madigan NN, Flügel A, Dockery P, O'Brien T, Howard L, Windebank AJ, Barry FP

The aim of this study was to assess the feasibility of transplanting mesenchymal stem cells (MSCs), genetically modified to express glial-derived neurotrophic factor (GDNF), to the contused rat spinal cord, and to subsequently assess their neural differentiation potential. MSCs expressing green fluorescent protein were transduced with a retroviral vector to express the neurotrophin GDNF. The transduction protocol was optimized by using green fluorescent protein-expressing retroviral constructs; approximately 90% of MSCs were transduced successfully after G418 selection. GDNF-transduced MSCs expressed the transgene and secreted growth factor into the media (approximately 12 ng/500,000 cells secreted into the supernatant 2 weeks after transduction). Injuries were established using an impactor device, which applied a given, reproducible force to the exposed spinal cord. GDNF-expressing MSCs were transplanted rostral and caudal to the site of injury. Spinal cord sections were analyzed 2 and 6 weeks after transplantation. We demonstrate that GDNF-transduced MSCs engraft, survive, and express the therapeutic gene up to 6 weeks posttransplantation, while maintaining an undifferentiated phenotype. In conclusion, transplanted MSCs have limited capacity for the replacement of neural cells lost as a result of a spinal cord trauma. However, they provide excellent opportunities for local delivery of neurotrophic factors into the injured tissue. This study underlines the therapeutic benefits associated with cell transplantation and provides a good example of the use of MSCs for gene delivery.

PMID: 19335061 [PubMed - indexed for MEDLINE]


Science and Research in Academic Plastic Surgery in Germany.
December 24, 2009 at 6:02 am

Science and Research in Academic Plastic Surgery in Germany.

Handchir Mikrochir Plast Chir. 2009 Dec;41(6):359-363

Authors: Giunta RE, Machens HG

Plastic surgery has passed through a very positive evolution in the last decades on the solid fundament of constantly developing academic plastic surgery. Aim of this paper is an objective evaluation of the current status of academic plastic surgery regarding research topics, currently available ressources and scientific outcome based on a questionnaire. The return rate of the questionnaire in academic departments was 92%. Main topics in research besides wound healing were topics from regenerative medicine such as tissue engineering, biomaterials, genetherapy and angiogenesis with the main focus on skin and fat tissues. In the past five years a total of 25 million Euros of third party research grants were raised. Research relied mainly on interdisciplinary research facilities. Regarding the scientific outcome more than 200 scientific papers were published in basic science research journals having an impactfactor higher than two. These results clearly demonstrate that plastic surgery is scientifically highly productive in academic surroundings where independent departments are established. Considering that independent units of plastic surgery exist in a relatively small number of all 36 university hospitals in germany, it has to be claimed for further independent departments so to provide adequate research facilities for further evolution of academic plastic surgery.

PMID: 20029742 [PubMed - as supplied by publisher]


Specific fibrinogen and thrombin concentrations promote neuronal rather than glial growth when primary neural cells are seeded within plasma-derived fibrin gels.
December 24, 2009 at 6:02 am

Specific fibrinogen and thrombin concentrations promote neuronal rather than glial growth when primary neural cells are seeded within plasma-derived fibrin gels.

Tissue Eng Part A. 2009 Dec 22;

Authors: Mooney RG, Tawil BJ, Mahoney MJ

Fibrin gels are attractive scaffolds useful for neural tissue engineering applications. The objective of this work was to investigate the apoptotic activity, survival, proliferation, and differentiation of a mixed population of primary neural cells composed of neurons and multipotent precursor cells when cultured in fibrin gels prepared with varying concentrations of fibrinogen (5 mg/ml - 25 mg/ml fibrinogen) and thrombin (1 U/ml - 125 U/ml thrombin). Within all fibrin gel formulations tested, the level of apoptosis on day 1 was low and cell survival was equivalent to levels in monolayer culture (67%). Proliferation in gels made from 5 mg/ml or 12.5 mg/ml fibrinogen was also similar to that observed in monolayer culture, though a lower proliferative response was observed in 25 mg/ml fibrinogen formulations. Relative to monolayer culture, cholinergic and dopaminergic neuronal presence was enhanced while glial cell growth was reduced in fibrin gel cultures. The extent to which levels were altered depended on fibrinogen and thrombin concentration. The findings here suggest the importance of fibrinogen and thrombin concentration in differentially regulating the growth and composition of neural cell populations and are of importance for neural tissue engineering strategies focused on the development of implantable scaffolds for treating neurodegenerative disorders.

PMID: 20028220 [PubMed - as supplied by publisher]


Dynamic mechanical loading enhances functional properties of tissue engineered cartilage using mature canine chondrocytes.
December 24, 2009 at 6:02 am

Dynamic mechanical loading enhances functional properties of tissue engineered cartilage using mature canine chondrocytes.

Tissue Eng Part A. 2009 Dec 22;

Authors: Bian L, Fong JV, Lima EG, Stoker AM, Ateshian GA, Cook JL, Hung CT

Objective: The concept of cartilage functional tissue engineering (FTE) has promoted the use of physiologic loading bioreactor systems to cultivate engineered tissues with load-bearing properties. Prior studies have demonstrated that culturing agarose constructs seeded with primary bovine chondrocytes from immature joints, and subjected to dynamic deformation, produced equilibrium compressive properties and proteoglycan content matching the native tissue. In the process of translating these results to an adult canine animal model, it was found that protocols previously successful with immature bovine primary chondrocytes did not produce the same successful outcome when using adult canine primary chondrocytes. The objective of this study was to assess the efficacy of a modified FTE protocol using adult canine chondrocytes seeded in agarose hydrogel and subjected to dynamic loading. Method: Two modes of dynamic loading were applied to constructs using custom bioreactors: unconfined axial compressive deformational loading (DL, 1Hz, 10% deformation) or sliding contact loading (SLIDE, 0.5Hz, 10% deformation). Loading for 3 hours daily was initiated on day 0, 14 or 28 (DL0, DL14, DL28 and Slide14). Results: Constructs with applied loading (both DL and Slide) exhibited significant increases in Young's modulus compared to free-swelling (FS) control as early as day 28 in culture, p<0.05. However, glycosaminoglycan, collagen and DNA content were not statistically different among the various groups. The modulus values attained for engineered constructs compare favorably with (and exceed in some cases) those of native canine knee (patella groove and condyle) cartilage. Conclusion: Our findings successfully demonstrate a functional tissue engineering strategy incorporating clinically-relevant, adult chondrocytes and gel scaffold for engineering cartilage replacement tissue. These results, using continuous growth factor supplementation, are in contrast to our previously reported studies with immature chondrocytes where the sequential application of dynamic loading after transient TGF-beta3 application was found to be a superior culture protocol. Sliding which simulates aspects of joint articulation has shown promise in promoting engineered tissue development and provides an alternative option for functional tissue engineering of cartilage constructs to be further explored.

PMID: 20028219 [PubMed - as supplied by publisher]


Effect of Nell-1 delivery on chondrocyte proliferation and cartilaginous extracellular matrix deposition.
December 24, 2009 at 6:02 am

Effect of Nell-1 delivery on chondrocyte proliferation and cartilaginous extracellular matrix deposition.

Tissue Eng Part A. 2009 Dec 22;

Authors: Lee M, Siu RK, Ting K, Wu B

Cartilage tissue engineering using chondrogenic growth factors is an attractive strategy to promote cartilage repair. Bone morphogenetic proteins (BMPs) have been widely studied for their application in cartilage repair. However, functional heterogeneity of BMPs and unpredictable effects such as cyst formation may limit their therapeutic use. Thus, the use of alternative growth factors with greater osteochondral specificity may be advantageous for cartilage regeneration. Nell-1 [Nel-like molecule-1; Nel (a protein strongly expressed in neural tissue encoding epidermal growth factor like domain)] is a novel growth factor believed to specifically target cells committed to the osteochondral lineage. Mutation of the Nell-1gene has been shown to disrupt normal cartilage growth and development in rodents. This study investigates the chondrogenic potential of recombinant human Nell-1 protein in a three-dimensional (3D) alginate hydrogel microenvironment containing rabbit chondrocytes. To provide controlled delivery and maximize biological efficiency, Nell-1 was incorporated in chitosan microparticles. Over 42 days of culture, chondrocyte proliferation and cluster formation was significantly enhanced by Nell-1 in a dose dependent manner. Furthermore, the clusters formed in the presence of Nell-1 contained more type II collagen and glycosaminoglycans than clusters formed within Nell-free control gels. These findings demonstrate the ability of Nell-1 to promote chondrocyte proliferation and deposition of cartilage specific extracellular matrix materials.

PMID: 20028218 [PubMed - as supplied by publisher]


Stem cells may improve the adaptability of dental implants.
December 24, 2009 at 6:02 am

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Stem cells may improve the adaptability of dental implants.

J Mich Dent Assoc. 2009 Sep;91(9):26

Authors: Foe D

PMID: 19835211 [PubMed - indexed for MEDLINE]


Treatment of human infrabony periodontal defects by grafting human cultured periosteum sheets combined with platelet-rich plasma and porous hydroxyapatite granules: case series.
December 24, 2009 at 6:02 am

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Treatment of human infrabony periodontal defects by grafting human cultured periosteum sheets combined with platelet-rich plasma and porous hydroxyapatite granules: case series.

J Int Acad Periodontol. 2009 Jul;11(3):206-13

Authors: Okuda K, Yamamiya K, Kawase T, Mizuno H, Ueda M, Yoshie H

In a previous publication from our research group we reported that a combination of platelet-rich plasma (PRP) and hydroxyapatite (HA) granules resulted in a significantly favorable clinical result in treating osseous defects. The aim of this study is to evaluate the clinical response of human cultured periosteum (HCP) sheets in combination with autologous PRP and HA granules used as a tissue-engineered grafting material for the treatment of infrabony defects in three female patients. Periosteum cell specimens were harvested from the mandible of each patient and cultured for six weeks until HCP sheets were formed. The periodontal surgery fully exposed the osseous defects and debridement and root planing were carried out. The coagulated PRP and HA mixture were placed into the osseous defects, and then they were covered with autologous HCP sheets. Six months post-surgery there were gains in clinical attachment and radiographic evidence of infrabony osseous fill. It may be concluded that HCP sheets combined with PRP and HA granules showed significant promise for treating human periodontal infrabony defects. A factor contributing to these favorable clinical results would be the presence of osteogenic cells in the HCP sheets, which provided greater regeneration potential.

PMID: 19753798 [PubMed - indexed for MEDLINE]


Planar biaxial behavior of fibrin-based tissue-engineered heart valve leaflets.
December 24, 2009 at 6:02 am

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Planar biaxial behavior of fibrin-based tissue-engineered heart valve leaflets.

Tissue Eng Part A. 2009 Oct;15(10):2763-72

Authors: Robinson PS, Tranquillo RT

To design more effective tissue-engineered heart valve replacements, the replacement tissue may need to mimic the biaxial stress-strain behavior of native heart valve tissue. This study characterized the planar biaxial properties of tissue-engineered valve leaflets and native aortic valve leaflets. Fibrin-based valve equivalent (VE) and porcine aortic valve (PAV) leaflets were subjected to incremental biaxial stress relaxation testing, during which fiber alignments were measured, over a range of strain ratios. Results showed that VE leaflets exhibited a modulus and fiber reorientation behavior that correlated with strain ratio. In contrast, PAV leaflets maintained their relaxed modulus and fiber alignment when exposed to nonequibiaxial strain, but exhibited changes in stress relaxation. In uniaxial and equi-biaxial tension, there were few observed differences in relaxation behavior between VE and PAV leaflets, despite differences in the modulus and fiber reorientation. Likewise, in both tissues there was similar relaxation response in the circumferential and radial directions in biaxial tension, despite different moduli in these two directions. This study presents some fundamental differences in the mechanical response to biaxial tension of fibrin-based tissue-engineered constructs and native valve tissue. It also highlights the importance of using a range of strain ratios when generating mechanical property data for valvular and engineered tissues. The data presented on the stress-strain, relaxation, and fiber reorientation of VE tissue will be useful in future efforts to mathematically model and improve fibrin-based tissue-engineered constructs.

PMID: 19368523 [PubMed - indexed for MEDLINE]


Electrostatic crosslinked in situ-forming in vivo scaffold for rat bone marrow mesenchymal stem cells.
December 24, 2009 at 6:02 am

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Electrostatic crosslinked in situ-forming in vivo scaffold for rat bone marrow mesenchymal stem cells.

Tissue Eng Part A. 2009 Oct;15(10):3201-9

Authors: Kim KS, Lee JY, Kang YM, Kim ES, Lee B, Chun HJ, Kim JH, Min BH, Lee HB, Kim MS

We herein formulated and characterized an in situ-forming gel consisting of sodium carboxymethylcellulose (CMC) and poly(ethyleneimine) (PEI) and examined its use as an in vivo scaffold for rat bone marrow stem cells (rBMSCs). The changes of zeta potential, size, and viscosity of CMC solutions with 0 to 30 wt% PEI confirmed the electrostatic interaction and temperature-dependence between anionic CMC and cationic PEI. The CMC/PEI solution produced an electrostatically crosslinked gel with a three-dimensional network structure. The CMC solution containing 10 wt% PEI transformed to a gel at temperatures greater than 35 degrees C and was chosen for subcutaneous injection into rats. The CMC/PEI (90/10) gel with pore structure acted as a suitable biocompatible substrate for the in vitro and in vivo attachment and proliferation of rBMSCs. As the CMC/PEI (90/10) solution with and without rBMSCs was injected into Fisher rats, it became a gel in the subcutaneous dorsum of the rats and maintained its shape even after 28 days in vivo. The injected rBMSCs survived in the CMC gel for 28 days. Injection of CMC/PEI gel alone induced macrophage accumulation in the host tissue and at the edge of the gel, whereas injection of CMC/PEI gel containing rBMSCs was associated with less macrophage accumulation, indicating immunosuppression by the transplanted rBMSCs. Our results collectively show that CMC/PEI gel could serve as an in situ-forming gel scaffold for entrapped rBMSCs in vivo.

PMID: 19366343 [PubMed - indexed for MEDLINE]


Effect of polymer molecular weight on the bone biological activity of biodegradable polymer/calcium phosphate cement composites.
December 24, 2009 at 6:02 am

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Effect of polymer molecular weight on the bone biological activity of biodegradable polymer/calcium phosphate cement composites.

Tissue Eng Part A. 2009 Oct;15(10):3183-91

Authors: Bodde EW, Habraken WJ, Mikos AG, Spauwen PH, Jansen JA

Previous studies demonstrated that the addition of biodegradable polymer microparticles to calcium phosphate (CaP) cement improves the cement's degradative behavior without affecting its handling characteristics, especially its injectability and moldability. We investigated the influence of molecular weight of polymeric microparticles included in CaP cement on implant degradation and bone formation in critical-sized defects. Forty rats received cranial defects filled with formulations of CaP cement and poly(DL-lactic-co-glycolic acid) (PLGA) microparticles. Microparticles consisted of 100% high- (HMW) or low-molecular-weight (LMW) PLGA or mixtures of these (25%, 50%, or 75%). Implantation time was 12 weeks. Porosity measurements showed that the 100% HMW group was significantly less porous than the other groups. Histology and histomorphometry revealed significantly greater implant degradation in the 100% LMW group. Defect bridging was mainly seen in the 75% and 100% LMW groups, with the highest amount of bone in the 100% LMW formulation. These results suggest that LMW PLGA microparticles are associated with better bone formation than HMW PLGA, which is most likely explained by the greater degradation of LMW PLGA microparticles. In conclusion, CaP cement composites with high percentages of LMW PLGA microparticles show good bone transductive behavior, with complete defect bridging. The 100% LMW group turned out to be the best formulation.

PMID: 19364281 [PubMed - indexed for MEDLINE]


Enhancement of embryonic stem cell differentiation promoted by avian chorioallantoic membranes.
December 24, 2009 at 6:02 am

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Enhancement of embryonic stem cell differentiation promoted by avian chorioallantoic membranes.

Tissue Eng Part A. 2009 Oct;15(10):3193-200

Authors: Talavera-Adame D, Dafoe DC, Ng TT, Wachsmann-Hogiu S, Castillo-Henkel C, Farkas DL

Avian chorioallantoic membrane (CAM) has been used as a model to explore angiogenesis and to study the microvasculature of transplanted tissues. Because CAM provides a vascular bed, cells can be implanted, and their development can be monitored and modified. We used the CAM model to study the differentiation process of embryoid bodies (EBs) derived from mouse embryonic stem cells (ESCs) influenced by the CAM vascular bed. After EBs were incubated in CAM for 5 days, they underwent further differentiation and became tissue masses (TMs) of different morphologies from those that grew outside CAM. Immunohistochemical analysis of TMs demonstrated tissue-specific markers such as neurofilament light, CD34, collagen IV, cardiac myosin heavy chain (MHC), and cardiotin. Differentiated mouse blood vessels stained with anti-CD31 were found within the TMs, as well as blood vessels stained positive for QH1 and QCPN, markers for quail endothelial cells and perinuclear quail antigen, respectively. Quail erythrocytes inside mouse blood vessels suggested a connection between existing quail vessels and blood vessels growing inside the TMs as a result of EB differentiation. Therefore, CAM could be a suitable model to trigger and study the differentiation of EBs in close interaction with surrogate quail blood vessels.

PMID: 19364272 [PubMed - indexed for MEDLINE]


Engineered extracellular matrices modulate the expression profile and feeder properties of bone marrow-derived human multipotent mesenchymal stromal cells.
December 24, 2009 at 6:02 am

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Engineered extracellular matrices modulate the expression profile and feeder properties of bone marrow-derived human multipotent mesenchymal stromal cells.

Tissue Eng Part A. 2009 Oct;15(10):3161-71

Authors: Seib FP, Müller K, Franke M, Grimmer M, Bornhäuser M, Werner C

The bone marrow harbors multipotent mesenchymal stromal cells (MSCs) that nurture hematopoietic stem cells (HSCs). The extracellular matrix (ECM) is an integral part of the bone marrow, and the aim of this study was therefore to examine the effect of engineered ECM substrates on MSC gene expression over time and to determine quantitatively the functional ability of ECM-cultured MSCs to support HSCs. ECMs were surface immobilized using thin films of maleic anhydride to covalently immobilize tropocollagen or fibrillar collagen type I to the substrate. Where indicated, collagen type I fibrils were supplemented with heparin or hyaluronic acid. All surfaces maintained MSC viability and supported cell expansion. Microarray analysis of MSCs cultured on engineered ECM substrates revealed that culture time, as well as substrate composition, significantly affected expression levels. Based on these studies, it was possible to predict the effect of these substrates on in vitro HSC clonogenicity and self-renewal. The ability to regulate the expression of stromal factors using engineered ECM is exciting and warrants further studies to identify the ECM components and combinations that maximize the expansion of clonogenic HSCs.

PMID: 19358630 [PubMed - indexed for MEDLINE]


CD133+ endothelial progenitor cells as a potential cell source for a bioartificial glomerulus.
December 24, 2009 at 6:02 am

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CD133+ endothelial progenitor cells as a potential cell source for a bioartificial glomerulus.

Tissue Eng Part A. 2009 Oct;15(10):3173-82

Authors: Vu DM, Masuda H, Yokoyama TA, Fujimura S, Kobori M, Ito R, Sawada K, Saito A, Asahara T

Development of a bioartificial glomerulus, a hemofilter in which the inner surface of hollow fibers is endothelialized, requires expandable, nonimmunogenic, antithrombogenic, and highly permeable endothelial cells. We used human umbilical cord blood CD133(+) endothelial progenitor cells (EPCs) to evaluate the feasibility of application of EPCs for bioartificial glomerulus. Numbers of adhered CD133(+) EPCs (adhered EPCs) was approximately 25 to 30 times as great in the expansion culture group as in the non-expansion group. Adhered EPCs had endothelial cell features, including the expression of CD31, Kinase domain region, von Willebrand factor, vascular endothelial-cadherin, positive for Ulex europeus agglutinin I staining, and up-take of acetylated low-density lipoprotein. Adhered EPCs secreted 6-keto-prostaglandin F(1alpha) identically to that secreted by human umbilical vein endothelial cells (HUVECs). The cells also expressed messenger RNA for phospholipase A(2), cyclooxygenase (COX)-1, COX-2, prostaglandin I(2) synthase, tissue plasminogen activator, and thrombomodulin (TM). TM protein in adhered EPCs properly activated protein C. Scanning electron microscopy revealed the suppression of platelet adhesion and aggregation on the surface of cell monolayer. Adhered EPCs treated with 50 microg/mL of cytochalasin B induced a larger diameter and a greater number of fenestrae, subsequently producing significantly more ultrafiltration than the non-treated cell. These results suggest that CD133(+) EPCs would potentially be applicable in bioartificial glomerulus.

PMID: 19358628 [PubMed - indexed for MEDLINE]


Amniotic fluid stem cells produce robust mineral deposits on biodegradable scaffolds.
December 24, 2009 at 6:02 am

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Amniotic fluid stem cells produce robust mineral deposits on biodegradable scaffolds.

Tissue Eng Part A. 2009 Oct;15(10):3129-38

Authors: Peister A, Deutsch ER, Kolambkar Y, Hutmacher DW, Guldberg RE

Insufficient availability of osteogenic cells limits bone regeneration through cell-based therapies. This study investigated the potential of amniotic fluid-derived stem (AFS) cells to synthesize mineralized extracellular matrix within porous medical-grade poly-varepsilon-caprolactone (mPCL) scaffolds. The AFS cells were initially differentiated in two-dimensional (2D) culture to determine appropriate osteogenic culture conditions and verify physiologic mineral production by the AFS cells. The AFS cells were then cultured on 3D mPCL scaffolds (6-mm diameter x 9-mm height) and analyzed for their ability to differentiate to osteoblastic cells in this environment. The amount and distribution of mineralized matrix production was quantified throughout the mPCL scaffold using nondestructive micro computed tomography (microCT) analysis and confirmed through biochemical assays. Sterile microCT scanning provided longitudinal analysis of long-term cultured mPCL constructs to determine the rate and distribution of mineral matrix within the scaffolds. The AFS cells deposited mineralized matrix throughout the mPCL scaffolds and remained viable after 15 weeks of 3D culture. The effect of pre-differentiation of the AFS cells on the subsequent bone formation in vivo was determined in a rat subcutaneous model. Cells that were pre-differentiated for 28 days in vitro produced seven times more mineralized matrix when implanted subcutaneously in vivo. This study demonstrated the potential of AFS cells to produce 3D mineralized bioengineered constructs in vitro and in vivo and suggests that AFS cells may be an effective cell source for functional repair of large bone defects.

PMID: 19344289 [PubMed - indexed for MEDLINE]


Preadipocytes stimulate ductal morphogenesis and functional differentiation of human mammary epithelial cells on 3D silk scaffolds.
December 24, 2009 at 6:02 am

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Preadipocytes stimulate ductal morphogenesis and functional differentiation of human mammary epithelial cells on 3D silk scaffolds.

Tissue Eng Part A. 2009 Oct;15(10):3087-98

Authors: Wang X, Zhang X, Sun L, Subramanian B, Maffini MV, Soto A, Sonnenschein C, Kaplan DL

Epithelial-mesenchymal interactions play an important role in regulating normal tissue development as well as tumor development for the mammary gland, but much is yet to uncover to reach a full understanding of their complexity. To address this issue, the establishment of relevant, surrogate, three-dimensional (3D) human tissue culture models is essential. In the present study, a novel 3D coculture system was developed to study the interactions between human mammary epithelial cells (MCF10A) and adipocytes, a prominent stromal cell type in native breast tissue. The MCF10A cells were cultured within a mixture of Matrigel and collagen in 3D porous silk scaffolds with or without predifferentiated human adipose-derived stem cells (hASCs). The presence of hASCs inhibited MCF10A cell proliferation, induced both alveolar and ductal morphogenesis, and enhanced their functional differentiation as evidenced by histology and functional analysis. The alveolar structures formed by cocultures exhibited proper, immature polarity when compared with native breast tissue. In contrast, only alveolar structures with reverted polarity were observed in the MCF10A monocultures. The effect of ductal morphogenesis in cocultures may correlate to hepatocyte growth factor secreted by the predifferentiated hASCs, based on results from a cytokine blocking assay. Taken together, this in vitro coculture model on silk scaffolds effectively reconstitutes a physiologically relevant 3D microenvironment for epithelial cells and stromal cells and provides a useful system to study tissue organization and epithelial morphogenesis in normal or diseased breast development.

PMID: 19338449 [PubMed - indexed for MEDLINE]


Surface-phosphorylated copolymer promotes direct bone bonding.
December 24, 2009 at 6:02 am

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Surface-phosphorylated copolymer promotes direct bone bonding.

Tissue Eng Part A. 2009 Oct;15(10):3061-9

Authors: Gopalakrishnanchettiyar SS, Mohanty M, Kumary TV, Valappil MP, Parameshwaran R, Varma HK

The bone bonding potential of surface-phosphorylated poly (2-hydroxyethyl methacrylate-co-methyl methacrylate) [poly (HEMA-co-MMA)] has been investigated and compared with commercially available poly (methyl methacrylate) bone cement (CMW1 radiopaque, Depuy; Johnson & Johnson, Blackpool, Lancashire, England, United Kingdom) as control. Poly (HEMA-co-MMA) is synthesized by free radical-initiated copolymerization and surface functionalized by phosphorylation. The X-ray photoelectron spectroscopy confirms the presence of surface-bound phosphate groups on poly (HEMA-co-MMA). The surface-phosphorylated poly (HEMA-co-MMA) promotes in vitro biomineralization, cell viability, cell adhesion, and expression of bone-specific markers such as osteocalcin and alkaline phosphatase. The bone implantation study performed in rabbits as per ISO 10993-6; 1994 (E) shows that surface-phosphorylated poly (HEMA-co-MMA) elicits bone bonding and new bone formation. New woven bone trabeculae are formed at the defect site of surface-phosphorylated poly (HEMA-co-MMA) within 1 week, while for control sample, inflammatory cells--predominantly, macrophages, fibroblasts, and fibrocytes--are present at the cortical margins around the defect. The 4 and 12 weeks postimplantation results show that the major part of the defects around the surface-phosphorylated poly (HEMA-co-MMA) implant is bridged with new woven bone, with significant remodeling (evident from resorption bays) along both the margins of the defect, but for control implants, the defects are only partially closed, with slight remodeling along the margins, but most of them are separated by fibrous tissue.

PMID: 19338435 [PubMed - indexed for MEDLINE]

 

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