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Human Wharton’s Jelly Mesenchymal Stem Cells Plasticity Augments Scar-Free Skin Wound Healing with Hair Growth  [PDF]
Vikram Sabapathy, Balasubramanian Sundaram, Sreelakshmi VM, Pratheesh Mankuzhy, Sanjay Kumar
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0093726
Abstract: Human mesenchymal stem cells (MSCs) are a promising candidate for cell-based transplantation and regenerative medicine therapies. Thus in the present study Wharton’s Jelly Mesenchymal Stem Cells (WJ-MSCs) have been derived from extra embryonic umbilical cord matrix following removal of both arteries and vein. Also, to overcome the clinical limitations posed by fetal bovine serum (FBS) supplementation because of xenogeneic origin of FBS, usual FBS cell culture supplement has been replaced with human platelet lysate (HPL). Apart from general characteristic features of bone marrow-derived MSCs, wharton jelly-derived MSCs have the ability to maintain phenotypic attributes, cell growth kinetics, cell cycle pattern, in vitro multilineage differentiation plasticity, apoptotic pattern, normal karyotype-like intrinsic mesenchymal stem cell properties in long-term in vitro cultures. Moreover, the WJ-MSCs exhibited the in vitro multilineage differentiation capacity by giving rise to differentiated cells of not only mesodermal lineage but also to the cells of ectodermal and endodermal lineage. Also, WJ-MSC did not present any aberrant cell state upon in vivo transplantation in SCID mice and in vitro soft agar assays. The immunomodulatory potential assessed by gene expression levels of immunomodulatory factors upon exposure to inflammatory cytokines in the fetal WJ-MSCs was relatively higher compared to adult bone marrow-derived MSCs. WJ-MSCs seeded on decellularized amniotic membrane scaffold transplantation on the skin injury of SCID mice model demonstrates that combination of WJ-MSCs and decellularized amniotic membrane scaffold exhibited significantly better wound-healing capabilities, having reduced scar formation with hair growth and improved biomechanical properties of regenerated skin compared to WJ-MSCs alone. Further, our experimental data indicate that indocyanin green (ICG) at optimal concentration can be resourcefully used for labeling of stem cells and in vivo tracking by near infrared fluorescence non-invasive live cell imaging of labelled transplanted cells, thus proving its utility for therapeutic applications.
Angiogenesis in Rat Uterine Scar after Introduction af Autological Mesenchymal Stem Cells of Bone Marrow Origin  [PDF]
Igor Maiborodin, Natalia Yakimova, Vera Matveeva, Andrey Shevela, Elena Maiborodina, Ekaterina Pekareva, Olesya Tkachuk
Journal of Biomedical Science and Engineering (JBiSE) , 2011, DOI: 10.4236/jbise.2011.43023
Abstract: The results of injecting of autologic mesenchymal stem cells of bone marrow origin (AMSCBMO), transfected by the GFP gene, into the scar of rat uterine horns were studied by methods of light microscopy. After the introduction of AMSCBMO into the formed scar on the right (2 months after the ligation) large groups of blood vessels with cellular elements inside were present; groups like that were not found in the opposite side. Studying unstained sections under reflected ultraviolet light the sufficient bright luminescence in the endothelium and the external membrane of scar vessels was found in uterine horn only on the side of introduction of AMSCBMO. It was concluded that after the introduction of AMSCBMO into the scar tissue they form blood vessels by differentiation into endotheliocytes and pericytes. GFP gene expression not only in endothelium of vessels, but also in their external membrane indicates that differentiation of AMSCMBO is possible in endothelial and in pericytal directions.
Investigating the Role of P311 in the Hypertrophic Scar  [PDF]
Jianglin Tan,Xu Peng,Gaoxing Luo,Bing Ma,Chuan Cao,Weifeng He,Shunzong Yuan,Shirong Li,John A. Wilkins,Jun Wu
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0009995
Abstract: The mechanisms of hypertrophic scar formation are not fully understood. We previously screened the differentially expressed genes of human hypertrophic scar tissue and identified P311 gene as upregulated. As the activities of P311 in human fibroblast function are unknown, we examined the distribution of it and the effects of forced expression or silencing of expression of P311. P311 expression was detected in fibroblast-like cells from the hypertrophic scar of burn injury patients but not in peripheral blood mononuclear cells, bone marrow mesenchymal stem cells, epidermal cells or normal skin dermal cells. Transfection of fibroblasts with P311 gene stimulated the expression of alpha-smooth muscle actin (α-SMA), TGF-β1 and α1(I) collagen (COL1A1), and enhanced the contraction of fibroblast populated collagen lattices (FPCL). In contrast, interference of fibroblast P311 gene expression decreased the TGF-β1 mRNA expression and reduced the contraction of fibroblasts in FPCL. These results suggest that P311 may be involved in the pathogenesis of hypertrophic scar via induction of a myofibroblastic phenotype and of functions such as TGF-β1 expression. P311 could be a novel target for the control of hypertrophic scar development.
Biologicals and Fetal Cell Therapy for Wound and Scar Management  [PDF]
Nathalie Hirt-Burri,Albert-Adrien Ramelet,Wassim Raffoul,Anthony de Buys Roessingh,Corinne Scaletta,Dominique Pioletti,Lee Ann Applegate
ISRN Dermatology , 2011, DOI: 10.5402/2011/549870
Abstract: Few biopharmaceutical preparations developed from biologicals are available for tissue regeneration and scar management. When developing biological treatments with cellular therapy, selection of cell types and establishment of consistent cell banks are crucial steps in whole-cell bioprocessing. Various cell types have been used in treatment of wounds to reduce scar to date including autolog and allogenic skin cells, platelets, placenta, and amniotic extracts. Experience with fetal cells show that they may provide an interesting cell choice due to facility of outscaling and known properties for wound healing without scar. Differential gene profiling has helped to point to potential indicators of repair which include cell adhesion, extracellular matrix, cytokines, growth factors, and development. Safety has been evidenced in Phase I and II clinical fetal cell use for burn and wound treatments with different cell delivery systems. We present herein that fetal cells present technical and therapeutic advantages compared to other cell types for effective cell-based therapy for wound and scar management. 1. Introduction Cell-based therapies are penetrating gently into routine medical care and especially for wound management of skin. They offer the promise of repairing and/or replacing damaged tissue and restoring lost functionality, because ideally, they provide all of the factors necessary for wound healing. Several cell types and tissues have been proposed as starting material including autologous cells, adult stem cells including those derived from bone marrow, and adipose tissue, fetal cells, embryonic stem cells, platelets, and tissues from placental and amniotic fluid. These cell types are used for biological preparations in processing vaccines and medicinal, veterinary, and tissue engineering products [1–41]. As the literature and information is vast on cell-based therapies, this paper will concentrate on fetal cells as the choice in wound and scar management. Firstly, we will define differences between stem, and mesenchymal and fetal cells, as the literature is confusing with these terminologies, followed by a short review of fetal wound healing and associated processes. Importantly, cell choice and the technical specifications to outscale, stability, safety, and delivery are the major hurdles for development of biologicals for better wound treatments and scar management. Fetal skin cells present biological, technical, and therapeutic advantages lending towards possible routine cellular-based therapy for wound and scar management. All of these aspects
Topical delivery of mesenchymal stem cells and their function in wounds
J Michael Sorrell, Arnold I Caplan
Stem Cell Research & Therapy , 2010, DOI: 10.1186/scrt30
Abstract: The term mesenchymal stem cell (MSC) applies to adult fibroblast-like cells that differentiate along multiple mesenchymal pathways when exposed to proper stimuli [1,2]. These cells were identified first in murine bone marrow as plastic-adherent cells that formed fibroblast colony-forming units [3]. Other investigators began to adapt similar adherent populations as feeder layers for the propagation of various hematopoietic cell lineages [4]. This usage provided the first glimpse of the ability of MSCs to secrete potent bioactive factors that enabled them to regulate the function of other types of cells. This cellular regulatory capacity underlies the current notion that MSCs possess therapeutic potential to promote the healing of wounds and ischemic tissues [1,2]. This implies that MSCs can also function as therapeutic cells that modulate microenvironments and immunological competence, accelerate wound repair, and reduce fibrosis or scar formation or both. A number of recent studies have been translating this concept into experimental studies and further into clinical applications. To date, such applications include cardiovascular disease and myocardial infarction; brain and spinal cord injury; cartilage, bone, and tendon repair; Crohn disease; and skin wound repair [5-7].The phenotypic definition of MSCs has been hampered by the heterogeneity of this population [8,9]. Heterogeneity occurs among cells harvested from a single anatomic site and also occurs between cells harvested from different anatomic sites. Bone marrow and adipose tissues are currently the major sources for MSCs that are being used for preclinical and clinical studies. Adipose stromal cells (ASCs), though exhibiting differences, still share basic characteristics with bone marrow-derived cells [10-12]. As with MSCs, ASCs have been employed in animal wound repair models and in preliminary clinical studies such as myocardial infarction, Crohn disease, and skin wound repair [13]. Cells with MSC characte
Full-thickness tissue engineered skin constructed with autogenic bone marrow mesenchymal stem cells
LiJuan He,Xue Nan,YunFang Wang,LiDong Guan,CiXian Bai,ShuangShuang Shi,HongFeng Yuan,Lin Chen,DaQing Liu,XueTao Pei
Science China Life Sciences , 2007, DOI: 10.1007/s11427-007-0069-2
Abstract: To explore the feasibility of repairing clinical cutaneous deficiency, autogenic bone marrow mesenchymal stem cells (BMSCs) were isolated and differentiated into epidermal cells and fibroblasts in vitro supplemented with different inducing factors and biomaterials to construct functional tissueengineered skin. The results showed that after 72 h induction, BMSCs displayed morphologic changes such as typical epidermal cell arrangement, from spindle shape to round or oval; tonofibrils, melanosomes and keratohyaline granules were observed under a transmission electronic microscope. The differentiated cells expressed epidermal stem cell surface marker CK19 (59.66% ± 4.2%) and epidermal cells differentiation marker CK10. In addition, the induced epidermal cells acquired the anti-radiation capacity featured by lowered apoptosis following exposure to UVB. On the other hand, the collagen microfibrils deposition was noticed under a transmission electronic microscope after differentiating into dermis fibroblasts; RT-PCR identified collagen type I mRNA expression in differentiated cells; radioimmunoassay detected the secretion of interleukin-6 (IL-6) and interleukin-8 (IL-8) (up to 115.06 pg/mL and 0.84 ng/mL, respectively). Further in vivo implanting BMSCs with scaffold material shortened skin wound repair significantly. In one word, autogenic BMSCs have the potential to differentiate into epidermal cells and fibroblasts in vitro, and show clinical feasibility acting as epidermis-like and dermis-like seed cells in skin engineering.
Full-thickness tissue engineered skin constructed with autogenic bone marrow mesenchymal stem cells
HE LiJuan,NAN Xue,WANG YunFang,GUAN LiDong,BAI CiXian,SHI ShuangShuang,YUAN HongFeng,CHEN Lin,LIU DaQing &,PEI XueTao,

中国科学C辑(英文版) , 2007,
Abstract: To explore the feasibility of repairing clinical cutaneous deficiency, autogenic bone marrow mesenchymal stem cells (BMSCs) were isolated and differentiated into epidermal cells and fibroblasts in vitro supplemented with different inducing factors and biomaterials to construct functional tissueengineered skin. The results showed that after 72 h induction, BMSCs displayed morphologic changes such as typical epidermal cell arrangement, from spindle shape to round or oval; tonofibrils, melanosomes and keratohyaline granules were observed under a transmission electronic microscope. The differentiated cells expressed epidermal stem cell surface marker CK19 (59.66% ± 4.2%) and epidermal cells differentiation marker CK10. In addition, the induced epidermal cells acquired the anti-radiation capacity featured by lowered apoptosis following exposure to UVB. On the other hand, the collagen microfibrils deposition was noticed under a transmission electronic microscope after differentiating into dermis fibroblasts; RT-PCR identified collagen type I mRNA expression in differentiated cells; radioimmunoassay detected the secretion of interleukin-6 (IL-6) and interleukin-8 (IL-8) (up to 115.06 pg/mL and 0.84 ng/mL, respectively). Further in vivo implanting BMSCs with scaffold material shortened skin wound repair significantly. In one word, autogenic BMSCs have the potential to differentiate into epidermal cells and fibroblasts in vitro, and show clinical feasibility acting as epidermis-like and dermis-like seed cells in skin engineering. Supported by the Major Technology Program of Beijing Municipal Science & Technology Commission (Grant No. H060920050130) and the Major State Basic Research Development Program of China (Grant No. 2005CB522702)
The process of mesenchymal stem cells and their potential as cardiac therapeutics
骨髓间质干细胞修复受损心肌研究进展

LING Shu-kuan,LI Ying-hui,DAI Zhong-quan,YANG Fen,NIE Jie-lin,
凌树宽
,李莹辉,戴钟铨,杨芬,聂捷琳

中国生物工程杂志 , 2007,
Abstract: Bone marrow mesenchymal stem cells (MSCs), multipotent stem cells, can replicate as undifferentiated cells and have the potential to differentiate into different lineages of mesenchymal tissues, including bone, cartilage,endothelial, neural, smooth muscle, skeletal myoblasts, and cardiac myocyte cells. The ischemia-induced death of cardiomyocytes results in scar formation and reduced contractility of the ventricle. Several preclinical and clinical studies have supported the notion that MSCs therapy may be used for cardiac regeneration.When transplanted into the infracted heart, MSCs prevent deleterious remodeling and improve recovery, but the mechanism is not clear. In this work,we review evidence and new prospects that support the use of MSCs in cardiomyoplasty.
Evaluation of the Secretome Profile and Functional Characteristics of Human Bone Marrow Mesenchymal Stromal Cells-Derived Conditioned Medium Suggest Potential for Skin Rejuvenation  [PDF]
Sudha Balasubramanian, Charan Thej, Ankita Walvekar, Priyanka Swamynathan, Pawan K. Gupta, Raviraja N. Seetharam, Anish S. Majumdar
Journal of Cosmetics, Dermatological Sciences and Applications (JCDSA) , 2017, DOI: 10.4236/jcdsa.2017.71010
Abstract: Background/Purpose: Multipotent bone marrow-derived mesenchymal stromal cells (BMMSC) have been shown to possess the potential for tissue regeneration. The application of mesenchymal stromal cells (MSC)-derived growth factors and cytokines (GF/CKs) has been implicated for the repair and regeneration of the damaged skin that occurs due to aging and exposure to environmental stress factors. Methods: We have used both qualitative and quantitative measurements of the GF/CKs from the conditioned medium (CM) of a pooled population of BMMSC by antibody array analysis as well as by enzyme-linked immunosorbent assay (ELISA). Furthermore, the CM was also used in a variety of in vitro biological assays to measure its protective properties in human skin fibroblasts. Results: We have characterized the secretome of BMMSC by analyzing the composition of the CM using a forty-one growth factor array system. Thirteen of these GF/CK/extra cellular matrix (ECM)/ matrix metalloproteinases (MMP)-inhibitors in the CM were quantified owing to their involvement in skin repair cascade. In addition, we report that the BMMSC-CM was also able to protect dermal fibroblasts against tert-Butyl hydro peroxide (tbOH) induced oxidative stress and ultra violet B (UV-B) radiation induced cell damage. Conclusion: Based on the data presented here, we propose that BMMSC-derived CM may have the potential to promote health and rejuvenation of the aging skin.
Distant Mesenchymal Progenitors Contribute to Skin Wound Healing and Produce Collagen: Evidence from a Murine Fetal Microchimerism Model  [PDF]
Elke Seppanen, Edwige Roy, Rebecca Ellis, George Bou-Gharios, Nicholas M. Fisk, Kiarash Khosrotehrani
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0062662
Abstract: The contribution of distant and/or bone marrow-derived endogenous mesenchymal stem cells (MSC) to skin wounds is controversial. Bone marrow transplantation experiments employed to address this have been largely confounded by radiation-resistant host-derived MSC populations. Gestationally-acquired fetal MSC are known to engraft in maternal bone marrow in all pregnancies and persist for decades. These fetal cells home to damaged maternal tissues, mirroring endogenous stem cell behavior. We used fetal microchimerism as a tool to investigate the natural homing and engraftment of distant MSC to skin wounds. Post-partum wild-type mothers that had delivered transgenic pups expressing luciferase under the collagen type I-promoter were wounded. In vivo bioluminescence imaging (BLI) was then used to track recruitment of fetal cells expressing this mesenchymal marker over 14 days of healing. Fetal cells were detected in 9/43 animals using BLI (Fisher exact p = 0.01 versus 1/43 controls). These collagen type I-expressing fetal cells were specifically recruited to maternal wounds in the initial phases of healing, peaking on day 1 (n = 43, p<0.01). This was confirmed by detection of Y-chromosome+ve fetal cells that displayed fibroblast-like morphology. Histological analyses of day 7 wounds revealed vimentin-expressing fetal cells in dermal tissue. Our results demonstrate the participation of distant mesenchymal cells in skin wounds. These data imply that endogenous MSC populations are likely recruited from bone marrow to wounds to participate in healing.
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