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| Progress and prospects: graft-versus-host disease.
May 29, 2010 at 10:02 AM
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| | Progress and prospects: graft-versus-host disease. Gene Ther. 2010 May 27; Authors: Mastaglio S, Stanghellini MT, Bordignon C, Bondanza A, Ciceri F, Bonini C Graft-versus-host disease (GvHD) is one of the major complications of allogeneic hematopoietic stem cell transplantation, an otherwise highly effective therapeutic modality for patients affected by hematological diseases. The main inducers of GvHD are alloreactive donor T cells, which recognize host antigens presented by recipient cells. The critical role of lymphocytes in GvHD is well documented by the observation that T-cell depletion from the graft prevents GvHD. Unfortunately, the removal of donor lymphocytes from the graft increases the incidence of disease relapse and life-threatening infectious complications. Gene transfer technologies are promising tools to manipulate donor T-cell immunity to enforce graft-versus-tumor/graft-versus-infection while preventing or controlling GvHD. For this purpose, several cell and gene transfer approaches have been investigated at the preclinical level and implemented in clinical trials.Gene Therapy advance online publicati! on, 27 May 2010; doi:10.1038/gt.2010.83. PMID: 20508597 [PubMed - as supplied by publisher] | | |
| Mandibular reconstruction using nonvascularized autogenous bone grafting.
May 29, 2010 at 10:02 AM
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| | Mandibular reconstruction using nonvascularized autogenous bone grafting. Curr Opin Otolaryngol Head Neck Surg. 2010 May 24; Authors: Morrison A, Brady J PURPOSE OF REVIEW: This paper will discuss reconstruction of the mandible with autogenous nonvascularized bone grafting. New developments in this area will be investigated by reviewing the most recent literature on this topic as compared with other techniques currently employed. With the advances of vascularized free flap reconstruction it is important to investigate the indication for nonvascularized techniques. RECENT FINDINGS: Replacement of a portion of the mandibular bone is a common procedure for patients undergoing ablative cancer surgery or for infection as well as temporomandibular joint replacement secondary to disease or trauma. The subject of mandibular reconstruction has seen great advances in recent years with the advent of vascularized free tissue transfer. Other newer areas of mandibular replacement include tissue engineering and distraction osteogenesis. Traditional nonvascularized autogenous bone graft replacement can still play a vital role in r! ehabilitating these patients. SUMMARY: Although vascularized free flap reconstruction of mandibular defects has become the more common method of treating the postablative cancer surgery patient, there remain indications for nonvascularized reconstruction of mandibular defects as well as other techniques. PMID: 20508523 [PubMed - as supplied by publisher] | | |
| UV Absorbance of a Bioengineered Corneal Stroma Substitute in the 240-400 Range.
May 29, 2010 at 10:02 AM
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| | UV Absorbance of a Bioengineered Corneal Stroma Substitute in the 240-400 Range. Cornea. 2010 May 26; Authors: Ionescu AM, de la Cruz Cardona J, González-Andrades M, Alaminos M, Campos A, Hita E, Del Mar Pérez M PURPOSE:: To determine the UV absorbance of a bioengineered human corneal stroma construct based on fibrin and fibrin-agarose scaffolds in the 240-400 nm range. METHODS:: Three types of artificial substitutes of the human corneal stroma were developed by tissue engineering using fibrin and fibrin with 0.1% and 0.2% agarose scaffolds with human keratocytes immersed within. After 28 days of culture, the UV absorbance of each sample was determined using a spectrophotometer. The thickness of corneal stroma samples was determined by light microscope. RESULTS:: For all the 3 types of corneal stroma substitutes studied, the range of the UV absorbance values was similar to that of the native human corneal stroma, although the fibrin with 0.1% agarose stroma substitute had the best UV filtering properties. The higher UV absorbance of the artificial substitute of the human corneal stroma was in the UV-B and -A ranges suggesting that these artificial tissues could have poten! tial clinical usefulness and proper UV light-absorption capabilities. CONCLUSION:: Our data suggest that the bioengineered human corneal substitute of fibrin with 0.1% agarose is an effective absorber of harmful UV radiation and could therefore be potentially useful. PMID: 20508508 [PubMed - as supplied by publisher] | | |
| In Vivo and In Vitro Laser Confocal Microscopy to Diagnose Acanthamoeba Keratitis.
May 29, 2010 at 10:02 AM
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| | In Vivo and In Vitro Laser Confocal Microscopy to Diagnose Acanthamoeba Keratitis. Cornea. 2010 May 26; Authors: Shiraishi A, Uno T, Oka N, Hara Y, Yamaguchi M, Ohashi Y PURPOSE:: To determine the effectiveness of laser confocal microscopy in identifying Acanthamoeba cysts and trophozoites in the cornea of patients with Acanthamoeba keratitis (AK) and to evaluate its effectiveness in following AK after treatment. METHODS:: The corneas of 9 patients clinically diagnosed with AK were monitored periodically with the Heidelberg Retina Tomograph II-Rostock Cornea Module (HRT II-RCM) to search for Acanthamoeba cysts and trophozoites during the clinical course. RESULTS:: Seven of 9 patients had positive corneal smears, and 5 of 9 patients had positive laboratory cultures. HRT II-RCM demonstrated the presence of highly reflective polygonal shadows with lower reflective borders in the cornea of all patients. In 1 patient, a highly reflective pleomorphic shadow with small less-reflective areas was detected inside the cell. The former finding resembled the image of Acanthamoeba cysts in culture as observed by HRT II-RCM, and the latter obser! vation with that of Acanthamoeba trophozoites in culture. After treatment, the number of highly reflective inflammatory cells decreased and the number and morphology of the corneal epithelial cells with highly reflective nuclei recovered to normal levels. CONCLUSION:: These results indicate that in vivo laser confocal microscopy can be a useful method to make a diagnosis and to follow patients with AK. PMID: 20508505 [PubMed - as supplied by publisher] | | |
| Xenografted human amniotic membrane-derived mesenchymal stem cells are immunologically tolerated and transdifferentiated into cardiomyocytes.
May 29, 2010 at 10:02 AM
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| | Xenografted human amniotic membrane-derived mesenchymal stem cells are immunologically tolerated and transdifferentiated into cardiomyocytes. Circ Res. 2010 May 28;106(10):1613-23 Authors: Tsuji H, Miyoshi S, Ikegami Y, Hida N, Asada H, Togashi I, Suzuki J, Satake M, Nakamizo H, Tanaka M, Mori T, Segawa K, Nishiyama N, Inoue J, Makino H, Miyado K, Ogawa S, Yoshimura Y, Umezawa A Rationale: Amniotic membrane is known to have the ability to transdifferentiate into multiple organs and is expected to stimulate a reduced immunologic reaction. Objective: Determine whether human amniotic membrane-derived mesenchymal cells (hAMCs) can be an ideal allograftable stem cell source for cardiac regenerative medicine. Methods and Results: We established hAMCs. After cardiomyogenic induction in vitro, hAMCs beat spontaneously, and the calculated cardiomyogenic transdifferentiation efficiency was 33%. Transplantation of hAMCs 2 weeks after myocardial infarction improved impaired left ventricular fractional shortening measured by echocardiogram (34+/-2% [n=8] to 39+/-2% [n=11]; P<0.05) and decreased myocardial fibrosis area (18+/-1% [n=9] to 13+/-1% [n=10]; P<0.05), significantly. Furthermore hAMCs transplanted into the infarcted myocardium of Wistar rats were transdifferentiated into cardiomyocytes in situ and survived for more than 4 weeks after th! e transplantation without using any immunosuppressant. Immunologic tolerance was caused by the hAMC-derived HLA-G expression, lack of MHC expression of hAMCs, and activation of FOXP3-positive regulatory T cells. Administration of IL-10 or progesterone, which is known to play an important role in feto-maternal tolerance during pregnancy, markedly increased HLA-G expression in hAMCs in vitro and, surprisingly, also increased cardiomyogenic transdifferentiation efficiency in vitro and in vivo. Conclusions: Because hAMCs have a high ability to transdifferentiate into cardiomyocytes and to acquire immunologic tolerance in vivo, they can be a promising cellular source for allograftable stem cells for cardiac regenerative medicine. PMID: 20508201 [PubMed - in process] | | |
| Composite films of gelatin and hydroxyapatite/bioactive glass for tissue-engineering applications.
May 29, 2010 at 10:02 AM
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| | Composite films of gelatin and hydroxyapatite/bioactive glass for tissue-engineering applications. J Biomater Sci Polym Ed. 2010;21(8):1207-26 Authors: Gentile P, Chiono V, Boccafoschi F, Baino F, Vitale-Brovarone C, Vernè E, Barbani N, Ciardelli G Cross-linked gelatin/hydroxyapatite/bioactive glass (G/HA/CEL2) films with different compositions (100:0:0 (G1); 30:70:0 (G2); 30:0:70 (G3); 30:35:35 (G4) (%, w/w/w)) were prepared as scaffold materials for tissue-engineering applications, particularly in the field of bone repair. A bioactive glass with 45% SiO(2), 3% P(2)O(5), 26% CaO, 7% MgO, 15% Na(2)O and 4% K(2)O molar composition was selected (CEL2). Genipin was used as a cross-linker for the gelatin component. Samples were characterized in terms of their bioactivity, thermal properties, mechanical behaviour and cell compatibility. After only 3 days of incubation in simulated body fluid (SBF) at 37 degrees C, calcium phosphate crystals precipitated on G3 and G4 surfaces, due to the high CEL2 bioactivity. Cross-linking increased the thermal stability of the gelatine component as indicated by thermal analysis (denaturation temperature was 92.3 degrees C and 97.6 degrees C for not cross-linked and cross-linked ! gelatin, respectively). Furthermore, tensile modulus of samples increased with increasing the inorganic phase amount (from 4.72 +/- 0.23 MPa for G1 to 6.46 +/- 0.05 MPa for G4). The adhesion and proliferation of human primary osteoblasts on composite films was evaluated. Cell viability was high with respect to the control for all samples and the presence of hydroxyapatite exerted an important role in the ability of mineralization. PMID: 20507716 [PubMed - in process] | | |
| Regeneration of achilles' tendon: the role of dynamic stimulation for enhanced cell proliferation and mechanical properties.
May 29, 2010 at 10:02 AM
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| | Regeneration of achilles' tendon: the role of dynamic stimulation for enhanced cell proliferation and mechanical properties. J Biomater Sci Polym Ed. 2010;21(8):1173-90 Authors: Lee J, Guarino V, Gloria A, Ambrosio L, Tae G, Kim YH, Jung Y, Kim SH, Kim SH The tissue engineering of tendon was studied using highly elastic poly(L-lactide-co-epsilon-caprolactone) (PLCL) scaffolds and focusing on the effect of dynamic tensile stimulation. Tenocytes from rabbit Achilles tendon were seeded (1.0 x 10(6) cells/scaffold) onto porous PLCL scaffolds and cultured for periods of 2 weeks and 4 weeks. This was performed in a static system and also in a bioreactor equipped with tensile modulation which mimicked the environmental surroundings of tendons with respect to tensile extension. The degradation of the polymeric scaffolds during the culture was relatively slow. However, there was an indication that cells accelerated the degradation of PLCL scaffolds. The scaffold/cell adducts from the static culture exhibited inferior strength (at 2 weeks 350 kPa, 4 weeks 300 kPa) compared to the control without cells (at 2 weeks 460 kPa, 4 weeks 340 kPa), indicating that the cells contributed to the enhanced degradation. On the contrary, th! e corresponding values of the adducts from the dynamic culture (at 2 weeks 430 kPa, 4 weeks 370 kPa) were similar to, or higher than, those from the control. This could be explained by the increased quantity of cells and neo-tissues in the case of dynamic culture compensating for the loss in tensile strength. Compared with static and dynamic culture conditions, mechanical stimulation played a crucial role in the regeneration of tendon tissue. In the case of the dynamic culture system, cell proliferation was enhanced and secretion of collagen type I was increased, as evidenced by DNA assay and histological and immunofluorescence analysis. Thus, tendon regeneration, indicated by improved mechanical and biological properties, was demonstrated, confirming the effect of mechanical stimulation. It could be concluded that the dynamic tensile stimulation appeared to be an essential factor in tendon/ligament tissue engineering, and that elastic PLCL co-polymers could be very benefic! ial in this process. PMID: 20507714 [PubMed - in process] | | |
| Degradation Behavior and Biocompatibility of PEG/PANI-Derived Polyurethane Co-polymers.
May 29, 2010 at 10:02 AM
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| | Degradation Behavior and Biocompatibility of PEG/PANI-Derived Polyurethane Co-polymers. J Biomater Sci Polym Ed. 2010;21(8):1143-72 Authors: Luo YL, Nan YF, Xu F, Chen YS, Zhao P A series of polyurethane (PU) co-polymers with designable molecular weight between cross-linking dots was synthesized by a hydrogen transfer polymerization route from polyaniline (PANI), poly(ethylene glycol) (PEG), various curing agents and chain extenders using dibutyltin dilaurate as a catalyst. Their swelling, hydrophilicity, degradation and biocompatibility were inspected and assessed based on different degrees of polymerization of PANI and PEG, and their component proportion. Fourier transformation infrared spectrometry (FT-IR), (1)H-NMR spectroscopy, scanning electron microscopy (SEM), gel-permeation chromatography (GPC) and goniometry were used to characterize the structure and surface morphology of the synthesized PEG/PANI-based PU co-polymers, PU residues after degradation and degraded polymers at different time periods of hydrolysis. The thermal properties, aggregate structure and surface microstructure were examined by differential scanning calorimetry! (DSC), wide-angle X-ray diffraction (WAXD) and atomic force microscopy (AFM). Hemolysis, static platelet adhesion, dynamic clotting measurements and MTT assays were adopted to evaluate the hemo- or cytocompatibility. The experimental results indicated that these polymers exhibit various degrees of micro-phase separation, depending on the concentration and degree of polymerization of PANI, molecular weight of PEG, type of curing agent and chain extender, which further influence their swelling, hydrophilicity, degradable properties and biological performances in vitro. The incorporation of PANI and PANI* in co-polymers led to decreased thermal stability but slower decomposition rates than typical PEG-based PUs. The stress-strain tests showed that the as-prepared PU co-polymers possessed increased tensile strength and modulus, and decreased toughness in comparison with the blank PEG-based PU. These co-polymers are expected to find specific applications in tissue engineering o! r controlled drug release. PMID: 20507713 [PubMed - in process] | | |
| Electrospinning of matrigel to deposit a Basal lamina-like nanofiber surface.
May 29, 2010 at 10:02 AM
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| | Electrospinning of matrigel to deposit a Basal lamina-like nanofiber surface. J Biomater Sci Polym Ed. 2010;21(8):1081-101 Authors: de Guzman RC, Loeb JA, Vandevord PJ Schwann cell basal lamina is a nanometer-thin extracellular matrix layer that separates the axon-bound Schwann cells from the endoneurium of the peripheral nerve. It is implicated in the promotion of nerve regeneration after transection injury by allowing Schwann cell colonization and axonal guidance. Hence, it is desired to mimic the native basal lamina for neural tissue engineering applications. In this study, basal lamina proteins from BD Matrigel (growth factor-reduced) were extracted and electrospun to deposit nonwoven nanofiber mats. Adjustment of solute protein concentration, potential difference, air gap distance and flow rate produced a basal lamina-like construct with an average surface roughness of 23 nm and composed of 100-nm-thick irregular and relatively discontinuous fibers. Culture of embryonic chick dorsal root ganglion explants demonstrated that the fabricated nanofiber layer supported explant attachment, elongation of neurites, and migration of ! satellite Schwann cells in a similar fashion compared to electrospun collagen type-I fibers. Furthermore, the presence of nanorough surface features significantly increased the neurite spreading and Schwann cell growth. Sciatic nerve segment incubation also showed that the construct is promigratory to nerve Schwann cells. Results, therefore, suggest that the synthetic basal lamina fibers can be utilized as a biomaterial for induction of peripheral nerve repair. PMID: 20507710 [PubMed - in process] | | |
| Osteoblast Biocompatibility on Poly(octanediol citrate)/Sebacate Elastomers with Controlled Wettability.
May 29, 2010 at 10:02 AM
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| | Osteoblast Biocompatibility on Poly(octanediol citrate)/Sebacate Elastomers with Controlled Wettability. J Biomater Sci Polym Ed. 2010;21(8):1039-50 Authors: Djordjevic I, Szili EJ, Choudhury NR, Dutta N, Steele DA, Kumar S This work examines the biocompatibility of poly(octanediol citrate)/sebacate (p(OCS)) biodegradable polyester elastomers. The growth of human MG63 osteoblast-like cells was studied on p(OCS) films. Three types of p(OCS) films were synthesised simply by varying the concentrations of 1,8-octanediol (OD), citric acid (CA), and sebacic acid (SA) monomers at initial molar ratios of 1:1:0, 1:0.75:0.25 and 1:0.5:0.5. At these ratios, the p(OCS) films exhibit decreasing hydrophilicity as shown by the measured water contact angle values of 31, 41 and 64 degrees , respectively. For all the samples, no difference in cell growth was detected after 1 day of cell culture. However, after 4 days, the highest number of viable cells was detected on the p(OCS) film synthesised with the intermediate CA molar ratio of 0.75. This sample also contains the median concentration of surface carboxylic acid groups and hydrophilicity. Following long-term cell culture (18 days), a statisticall! y significant higher density of viable cells had grown on the p(OCS) films with SA molar ratios of 0.25 (P < 0.0001) and 0.5 (P = 0.002) in comparison to the material containing 100% CA and no SA. The work demonstrated that the performance of possible p(OCS) bone tissue engineering scaffolds could be improved by simply adjusting the molar ratios of CA and SA in the pre-polymer without any requirements for post-synthesis modification. PMID: 20507707 [PubMed - in process] | | |
| Haemodynamic Regulation of Gene Expression in Vascular Tissue Engineering.
May 29, 2010 at 10:02 AM
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| | Haemodynamic Regulation of Gene Expression in Vascular Tissue Engineering. Curr Vasc Pharmacol. 2010 May 28; Authors: Vara DS, Punshon G, Sales KM, Hamilton G, Seifalian AM Synthetic grafts, namely expanded polytetrafluoroethlene (ePTFE) and poly(ethylene terephthalate) (Dacron), used for cardiovascular bypass surgery are thrombogenic. Lining the inner lumen ("seeding") of synthetic grafts with endothelial cells (ECs) increases patency rates similar to those of autologous grafts (e.g. saphenous vein). The major drawback with seeding grafts is the retention of cells present on the graft after implantation in vivo, where large portions of cell wash off. Preconditioning the seeded EC monolayer with shear stress has been shown to promote the reorganisation of the EC cytoskeleton and production of extracellular matrix, resulting in higher EC retention after exposure to blood flow. Vascular ECs have a number of essential and complex roles. ECs synthesise and secrete vasoconstrictors, vasodilators, growth factors, fibrinolytic factors, cytokines, adhesion molecules, matrix proteins and mitogens that modulate many physiological processes suc! h as wound healing, hemostasis, vascular remodelling, inflammatory and immune responses. Vascular cells in vivo are exposed to hemodynamic forces created by the pulsatile flow of blood through the vessel. Due to their unique anatomical position, ECs are constantly exposed to shear stress forces and allow the vessel wall to adapt to changes by modulating EC structure and function. This review describes the mainly in vitro and in vivo studies used to define the molecular role hemodynamics have in vascular disease and its usage in developing tissue engineered vascular bypass grafts. PMID: 20507271 [PubMed - as supplied by publisher] | | |
| Artificial human tissues from cord and cord blood stem cells for multi-organ regenerative medicine: Viable alternatives to animal in vitro toxicology.
May 29, 2010 at 10:02 AM
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| | Artificial human tissues from cord and cord blood stem cells for multi-organ regenerative medicine: Viable alternatives to animal in vitro toxicology. Altern Lab Anim. 2010 May;38(2):183-92 Authors: Jurga M, Forraz N, McGuckin CP New medicinal products and procedures must meet very strict safety criteria before being applied for use in humans. The laboratory procedures involved require the use of large numbers of animals each year. Furthermore, such investigations do not always give an accurate translation to the human setting. Here, we propose a viable alternative to animal testing, which uses novel technology featuring human cord and cord blood stem cells. With over 130 million children born each year, cord and cord blood remains the most widely available alternative to the use of animals or cadaveric human tissues for in vitro toxicology. PMID: 20507188 [PubMed - in process] | | |
| Treatment viability of stem cells in ophthalmology.
May 29, 2010 at 10:02 AM
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| | Treatment viability of stem cells in ophthalmology. Curr Opin Ophthalmol. 2010 May;21(3):213-7 Authors: Jeganathan VS, Palanisamy M PURPOSE OF REVIEW: Adult ocular stem cells have the potential to restore vision in patients previously deemed incurable. This review summarizes strides in stem cell research and stumbling blocks that must be overcome to enable treatment viability in ophthalmology. RECENT FINDINGS: Stem/progenitor cells located in different regions of the eye are capable of differentiating enabling cell repopulation and tissue regeneration. At present, limbal epithelial stem transplantation is the sole ocular cell-based therapy being implemented into clinical practice. Research performed in animal models gives hope for using similar strategies to treat a wide range of ocular diseases in humans. The essential step toward successful therapeutic exploitation is to unravel regulators that control their cell proliferation and renewal pathways. The recently identified very small embryonic-like stem cells (VSEL-SCs) present in the bone marrow could potentially be harvested for regeneratio! n from cord blood via ex-vivo expansion and differentiation protocols SUMMARY: Although numerous impediments remain, the use of bioengineered stem cells is promising and may epitomize the future for replacement and regeneration of ocular tissues in various previously incurable ocular disorders. PMID: 20393292 [PubMed - indexed for MEDLINE] | | |
| Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells.
May 29, 2010 at 10:02 AM
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| | Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells. Nat Protoc. 2010;5(2):371-82 Authors: Aasen T, Belmonte JC The ease of generating induced pluripotent stem (iPS) cells, and possibly their properties after reprogramming, depends on the origin of the somatic cell starting population. Reprogramming of keratinocytes is both faster and more efficient compared with fibroblasts, although more care is required when isolating, culturing and infecting these cells. In this study, we describe detailed protocols using both feeder-dependent and defined serum- and feeder-free conditions for culturing human keratinocytes from foreskin samples and punch biopsies, as well as how to isolate keratinocytes from plucked hair. We further describe culture techniques and approaches to efficiently infect and reprogram these cells for the purpose of generating iPS cells. The procedure of deriving keratinocytes takes 10-14 d, whereas reprogramming and the appearance of iPS cell colonies that can be isolated and established requires another 3-4 weeks. PMID: 20134422 [PubMed - indexed for MEDLINE] | | |
| Efficacy of bone marrow-derived mesenchymal stem cells in the treatment of sclerodermatous chronic graft-versus-host disease: clinical report.
May 29, 2010 at 10:02 AM
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| | Efficacy of bone marrow-derived mesenchymal stem cells in the treatment of sclerodermatous chronic graft-versus-host disease: clinical report. Biol Blood Marrow Transplant. 2010 Mar;16(3):403-12 Authors: Zhou H, Guo M, Bian C, Sun Z, Yang Z, Zeng Y, Ai H, Zhao RC The success of treatment for sclerodermatous chronic graft-versus-host disease (ScGVHD) remains disappointing. The immunomodulatory ability of bone marrow (BM)-derived mesenchymal stem cells (MSCs) shows promise in treating GVHD, especially given its previous success in treating patients with acute GVHD (aGVHD). The potential efficacy and safety issues for treating cGVHD, particularly ScGVHD, remain to be clarified, however. Here, we report 4 patients with ScGVHD who received MSCs expanded ex vivo from unrelated donors by intra-BM injection. After MSC infusion, the ratio of helper T lymphocyte (Th) 1 cells to Th2 cells was dramatically reversed, with an increase in Th1 and a decrease in Th2 achieving a new balance. Correspondingly, symptoms gradually improved in all 4 patients. During the course of MSC treatment, the patients' vital signs and laboratory results remained normal. At the time of this report, none of the 4 patients had experienced recurrence of leukem! ia. Although this study alone cannot guarantee the application of MSCs in ScGVHD, our findings strongly suggest that this treatment is therapeutically practicable, with no detectable side effects. This approach may provide new insight into the clinical treatment of ScGVHD, with the aim of greatly increasing the survival rate in patients with leukemia who undergo allogeneic BM transplantation (BMT). PMID: 19925878 [PubMed - indexed for MEDLINE] | | | | 
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