골수기질세포 및 섬유아세포의 창상치유 촉진 성장인자 분비능 비교

김세현,한승규,윤태환,김우경 대한성형외과학회 2006 Archives of Plastic Surgery Vol.33 No.1

Cryopreserved fibroblast implants represent a major advancement for healing of chronic wounds. Bone marrow stromal cells, which include the mesenchymal stem cells, have a low immunity-assisted rejection and are capable of expanding profoundly in a culture media. Therefore, they have several advantages over fibroblasts in clinical use. The ultimate goal of this study was to compare the wound healing accelerating growth factor secretion of the bone marrow stromal cells with that of the fibroblasts and this pilot study particularly focuses on the growth factor secretion to accelerate wound healing. Bone marrow stromal cells and fibroblasts were isolated from the same patients and grown in culture. At 1, 3, and 5 days post-incubating, secretion of basic fibroblast growth factor(bFGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta(TGF-β) were compared. In TGF-β secretion fibroblasts showed 12~21% superior results than bone marrow stromal cells. In contrast, bFGF levels in the bone marrow stromal cells were 47~89% greater than that in fibroblasts. The VEGF levels of the bone marrow stromal cells was 7~12 fold greater than that of the fibroblasts. Our results suggest that the bone marrow stromal cells have great potential for wound healing accelerating growth factor secretion.

Current approaches in biomaterial-based hematopoietic stem cell niches

Bello, Alvin Bacero,Park, Hansoo,Lee, Soo-Hong Elsevier Science B.V. Amsterdam 2018 ACTA BIOMATERIALIA Vol.72 No.-

<P><B>Abstract</B></P> <P>Hematopoietic stem cells (HSCs) are multipotent progenitor cells that can differentiate and replenish blood and immune cells. While there is a growing demand for autologous and allogeneic HSC transplantation owing to the increasing incidence of hereditary and hematologic diseases, the low population of HSCs in cord-blood and bone marrow (the main source of HSCs) hinders their medical applicability. Several cytokine and growth factor-based methods have been developed to expand the HSCs <I>in vitro</I>; however, the expansion rate is low, or the expanded cells fail to survive upon engraftment. This is at least in part because the overly simplistic polystyrene culture substrates fail to fully replicate the microenvironments or niches where these stem cells live. Bone marrow niches are multi-dimensional, complex systems that involve both biochemical (cells, growth factors, and cytokines) and physiochemical (stiffness, O<SUB>2</SUB> concentration, and extracellular matrix presentation) factors that regulate the quiescence, proliferation, activation, and differentiation of the HSCs. Although several studies have been conducted on <I>in vitro</I> HSC expansion via 2D and 3D biomaterial-based platforms, additional work is required to engineer an effective biomaterial platform that mimics bone marrow niches. In this study, the factors that regulate the HSC <I>in vivo</I> were explained and their applications in the engineering of a bone marrow biomaterial-based platform were discussed. In addition, current approaches, challenges, and the future direction of a biomaterial-based culture and expansion of the HSC were examined.</P> <P><B>Statement of Significance</B></P> <P>Hematopoietic stem cells (HSC) are multipotent cells that can differentiate and replace the blood and immune cells of the body. However, <I>in vivo</I>, there is a low population of these cells, and thus their use in biotherapeutic and medical applications is limited (i.e., bone marrow transplantation). In this review, the biochemical factors (growth factors, cytokines, co-existing cells, ECM, gas concentrations, and differential gene expression) that may regulate the over-all fate of HSC, <I>in vivo,</I> were summarized and discussed. Moreover, different conventional and recent biomaterial platforms were reviewed, and their potential in generating a biomaterial-based, BM niche-mimicking platform for the efficient growth and expansion of clinically relevant HSCs <I>in-vitro</I>, was discussed.</P> <P><B>Graphical abstract</B></P> <P>On making an <I>in vitro</I> bone marrow (BM): The essential components. To make a bone marrow mimicking microenvironment for the growth and expansion of hematopoietic stem cells, several factors need to be taken into consideration. These factors include the BM’s in-vivo biochemical properties (growth factors hormones, co-existing cells, ECM, gas concentrations, regulated gene expression) and the biomaterial properties (type, dimension, topography, and mechanical strength).</P> <P>[DISPLAY OMISSION]</P>

Osteogenic and Angiogenic Potency of VEGF165-Transfected Canine Bone Marrow Mesenchymal Cells Combined with Coral Hydroxyapatite in Vitro

Zhang Quanyin,Zhang Jie,Chen Lin,Fan Yunjian,Long Jiazhen,Liu Shuguang 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.5

BACKGROUND: To explore the osteogenic and angiogenic potential of human vascular endothelial growth factor 165 (hVEGF165) gene-transfected canine bone marrow mesenchymal stem cells (BMSCs) combined with coral hydroxyapatite (CHA) scaffold. METHODS: We constructed a lentiviral vector and transfected canine BMSCs with the best multiplicity of infection. Osteogenesis was induced in the transfected groups (GFP-BMSCs group and hVEGF-BMSCs group) and non-transfected group (BMSCs group), followed by the evaluation of alkaline phosphatase (ALP) activity and alizarin red S staining. Cells from the three groups were co-cultured with CHA granules, respectively to obtain the tissue-engineered bone. MTT assay and fluorescence microscopy were employed to assess cell proliferation and adhesion. The expression of osteogenic and angiogenic related genes and proteins were evaluated at 7, 14, 21, and 28 days post osteoinduction in cell culture alone and cell co-culture with CHA, respectively using RT-PCR and ELISA. RESULTS: The hVEGF165 gene was transfected into BMSCs successfully. Higher ALP activity and more calcified nodules were found in the hVEGF-BMSCs group than in the control groups (p < 0.001). Cells attached and proliferated in CHA particles. Both cells cultured alone and cells co-culture with CHA expressed more osteogenic and angiogenic related genes and proteins in the hVEGF-BMSCs group compared to the GFP-BMSCs and BMSCs groups (p < 0.05). CONCLUSION: High expression of hVEGF165 in BMSCs potentially promote the osteogenic potential of BMSCs, and synergically drive the expression of other osteogenic and angiogenic factors. hVEGF-BMSCs co-cultured with CHA expressed more osteogenic and angiogenic related factors, creating a favorable microenvironment for osteogenesis and angiogenesis. Also, the findings have allowed for the construction of a CHA-hVEGF-BMSCs tissue-engineered bone. BACKGROUND: To explore the osteogenic and angiogenic potential of human vascular endothelial growth factor 165 (hVEGF165) gene-transfected canine bone marrow mesenchymal stem cells (BMSCs) combined with coral hydroxyapatite (CHA) scaffold. METHODS: We constructed a lentiviral vector and transfected canine BMSCs with the best multiplicity of infection. Osteogenesis was induced in the transfected groups (GFP-BMSCs group and hVEGF-BMSCs group) and non-transfected group (BMSCs group), followed by the evaluation of alkaline phosphatase (ALP) activity and alizarin red S staining. Cells from the three groups were co-cultured with CHA granules, respectively to obtain the tissue-engineered bone. MTT assay and fluorescence microscopy were employed to assess cell proliferation and adhesion. The expression of osteogenic and angiogenic related genes and proteins were evaluated at 7, 14, 21, and 28 days post osteoinduction in cell culture alone and cell co-culture with CHA, respectively using RT-PCR and ELISA. RESULTS: The hVEGF165 gene was transfected into BMSCs successfully. Higher ALP activity and more calcified nodules were found in the hVEGF-BMSCs group than in the control groups (p < 0.001). Cells attached and proliferated in CHA particles. Both cells cultured alone and cells co-culture with CHA expressed more osteogenic and angiogenic related genes and proteins in the hVEGF-BMSCs group compared to the GFP-BMSCs and BMSCs groups (p < 0.05). CONCLUSION: High expression of hVEGF165 in BMSCs potentially promote the osteogenic potential of BMSCs, and synergically drive the expression of other osteogenic and angiogenic factors. hVEGF-BMSCs co-cultured with CHA expressed more osteogenic and angiogenic related factors, creating a favorable microenvironment for osteogenesis and angiogenesis. Also, the findings have allowed for the construction of a CHA-hVEGF-BMSCs tissue-engineered bone.

Bone Marrow T Cells are Superior to Splenic T Cells to Induce Chimeric Conversion After Non-Myeloablative Bone Marrow Transplantation

( Hyun Sil Park ),( Seok Goo Cho ),( Min Jung Park ),( So Youn Min ),( Hong Seok Chang ),( Hee Je Kim ),( Seok Lee ),( Chang Ki Min ),( Jong Wook Lee ),( Woo Sung Min ),( Chun Choo Kim ),( Ho Youn Kim 대한내과학회 2009 The Korean Journal of Internal Medicine Vol.24 No.3

Background/Aims: The bone marrow functions not only as the primary B-lymphocyte-producing organ but also as a secondary lymphoid organ for CD4 and CD8 cell responses and a site of preferential homing and persistence for memory T cells. Bone marrow T (BM-T) cells are distinguished from peripheral blood T cells by surface phenotype, cytokine secretion profile, and immune functions. In this study, we evaluated the alloreactive potential of donor lymphocyte infusion (DLI) using BM-T cells in mixed chimerism compared to that using spleen T (SP-T) cells. Methods: Cells were prepared using established procedures. BM-T cells were obtained as a by-product of T-cell depletion in BM grafting and then cryopreserved for subsequent DLI. We performed DLI using BM-T cells in allogeneic mixed chimera mice on post-BMT day 21. Results: When the same dose of T cells, 5-10×105 (Thy1.2+), fractionated from BM and spleen were administered into mixed chimeras, the BM-T group showed complete chimeric conversion, with self-limited graftversus-host disease (GVHD) and no pathological changes. However, the SP-T group showed persistent mixed chimerism, with pathological signs of GVHD in the liver and intestine. Conclusions: Our results suggest that DLI using BM-T cells, even in small numbers, is more potent at inducing chimeric conversion in mixed chimerism than DLI using SP-T cells. Further study is needed to determine whether cryopreserved BM-T cells are an effective cell source for DLI to consolidate donor-dominant chimerism in clinical practice without concerns about GVHD. (Korean J Intern Med 2009;24:252-262)

Accelerate Bone Healing using Atelocollagen-based Solution Containing Bone Marrow-derived Mononuclear Cells(ASTEM-B) in Animal Bone Defect Model

( Jae Deog Jang ),( Hun Kim ),( Hyun Shin Park ),( Sang Hoon Woo ),( Jang Hoon Kim ),( Seon Ae Kim ),( Seok Jung Kim ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4

The osteogenic potential of autologous bone marrow derived mononuclear cells(MNCs) mixed with atelocollagen and hydroxy apatite when transplanted to bone defects was evaluated. 15 mm defect on radius were made in 27 NZW rabbits. The rabbits were divided into control, MATREX-B(atelocollagen and hydroxyl apatite mixture) and ASTEM-B(MATREX-B and MNCs mixture) groups as treatments. Each group was scheduled to be sacrificed at 3, 6 or 9 weeks after the operation and took an x-ray for radiological evaluation and tissue staining with Masson`s trichrome. At 9 weeks after the operation, ASTEM-B and MATREX-B treated group showed an excellent bone healing results. At 6 weeks after the operation, ASTEM-B treated group showed that most of the injected collagen gel formulation was converted to bone like tissue in the defect area. At 6 weeks after the operation, MATREX-B treated group showed an initial step in bone remodeling process. In case of control group, bone healing effect in defect area at 6 or 9 weeks after the operation showed a slight bone formation, but symptom of bone remodeling was not observed. In this research, atelocollagen gel formulation with autologous bone marrow derived mononuclear cells showed an accelerated bone healing using the rabbit model with critical bone defect.

Neural Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells: Applicability for Inner Ear Therapy

Lee, Jae-Hong,Kang, Won Kyung,Seo, Jae-Hyun,Choi, Mi-Yung,Lee, Yang Hyun,Kim, Hyo Min,Park, Kyoung-Ho 대한청각학회 2012 Journal of Audiology & Otology Vol.16 No.2

<P><B>Background and Objectives</B></P><P>Regeneration or replacement of damaged hair cells and neurons in the cochlea might be an ideal treatment for sensorineural hearing loss (SNHL). The purpose of the present study was to investigate whether mesenchymal stem cells (MSCs), derived from the bone marrow of rats, could differentiate into auditory hair cells and neurons.</P><P><B>Materials and Methods</B></P><P>The centrifuge gradient method was used to isolate MSCs from the bone marrow of rats. To confirm whether bone marrow-derived MSCs can differentiate into neuronal cells, culture medium with glial cell-derived neurotrophic factor, brain-derived neurotrophic factor and neurotrophin-3 for 14 days. In addition, immunofluorescence staining and RT-PCR were performed for characterization of the neurospheres and differentiated cells from 7 and 14 day cultures.</P><P><B>Results</B></P><P>The results showed that MSCs could differentiate into neuron-positive and hair cell-positive cells, using different compositions of growth factors. And RT-PCR result was identified high or low of gene expression all these differentiated cells.</P><P><B>Conclusions</B></P><P>Rat bone marrow-derived MSCs differentiated into neuronal progenitor cells. These cells might be useful for the treatment of SNHL.</P>

조혈모세포이식술이 골수기질세포의 기원 및 조골세포로의 증식 및 분화에 미치는 효과

강무일,조성원,오은숙,백기현,이원영,오기원,김혜수,한제호,윤건호,차봉연,이광우,손호영,강성구,김춘추 대한내분비학회 2000 Endocrinology and metabolism Vol.15 No.4·5

Enhanced bone formation by marrow-derived endothelial and osteogenic cell transplantation

Kim, Sang-Soo,Park, Min Sun,Cho, Seung-Woo,Kang, Sun Woong,Ahn, Kang-Min,Lee, Jong-Ho,Kim, Byung-Soo Wiley Subscription Services, Inc., A Wiley Company 2010 Journal of biomedical materials research. Part A Vol.a92 No.1

<P>Bone marrow-derived osteogenic cells can regenerate bone tissuesin vivo. The aim of the present study is to determine whether the cotransplantation of bone marrow-derived endothelial-like cells (BMECs) enhances bone regeneration by bone marrow-derived osteogenic cell (BMOC) transplantation in osseous defects. Canine bone marrow cells were differentiated separately into BMECs and BMOCs. Using apatite-coated poly(lactide-co-glycolide)/hydroxyapatite composite scaffolds as cell delivery vehicles, BMOCs were transplanted with or without BMECs into critical-sized calvarial defects in immunodeficient mice. Histological analyses, microcomputed tomography, and soft X-ray were performed to assess mineralized bone formation at 8 weeks. Cotransplantation of BMECs and BMOCs resulted in greater bone formation than transplantation of BMOCs alone. There was a significant (p < 0.05) increase in bone formation area following cotransplantation (30.8% ± 2.5%), compared with transplantation of BMOCs alone (15.3% ± 1.9%). These results demonstrate that the cotransplantation of BMECs enhances bone regeneration mediated by BMOC transplantation in osseous defects. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010</P>

Osteogenic Potency of Nacre on Human Mesenchymal Stem Cells

Green, David W.,Kwon, Hyuk-Jae,Jung, Han-Sung Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.3

Nacre seashell is a natural osteoinductive biomaterial with strong effects on osteoprogenitors, osteoblasts, and osteoclasts during bone tissue formation and morphogenesis. Although nacre has shown, in one study, to induce bridging of new bone across large non-union bone defects in 8 individual human patients, there have been no succeeding human surgical studies to confirm this outstanding potency. But the molecular mechanisms associated with nacre osteoinduction and the influence on bone marrow-derived mesenchymal stem cells (BMSC's), skeletal stem cells or bone marrow stromal cells remain elusive. In this study we highlight the phenotypic and biochemical effects of Pinctada maxima nacre chips and the global nacre soluble protein matrix (SPM) on primary human bone marrow-derived stromal cells (hBMSCs) in vitro. In static co-culture with nacre chips, the hBMSCs secreted Alkaline phosphatase (ALP) at levels that exceeded bone morphogenetic protein (rhBMP-2) treatment. Concentrated preparation of SPM applied to Stro-1 selected hBMSC's led to rapid ALP secretions, at concentrations exceeding the untreated controls even in osteogenic conditions. Within 21 days the same population of Stro-1 selected hBMSCs proliferated and secreted collagens I-IV, indicating the premature onset of an osteoblast phenotype. The same SPM was found to promote unselected hBMSC differentiation with osteocalcin detected at 7 days, and proliferation increased at 7 days in a dose-dependent manner. In conclusion, nacre particles and nacre SPM induced the early stages of human bone cell differentiation, indicating that they may be promising soluble factors with osteoinductive capacity in primary human bone cell progenitors such as, hBMSC's.

골수 유래 기질 줄기세포의 탐식작용 매개성 케모카인 수용체 발현 연구

정영신,변향민,신지영,김정목,정형민,오유경 한국약제학회 2003 Journal of Pharmaceutical Investigation Vol.33 No.4

To design gene delivery systems which can deliver higher amounts of genes into stem cells, we studied the expression of receptors involved in the receptor-mediated endocytosis of bone marrow stromal stem cells. Bone marrow was isolated from ICR mice, and bone marrow stromal stem cells were isolated based on their plastic adherence property. Several cultrure conditions were screened for effective and continuous culture of marrow stromal stem cells. MesenCult medium was finally used to cultivate marrow stromal stem cells in vitro. As candidate receptors, various chemokine receptors were studied. Both bone marrow cells and marrow-derived stromal stem cells showed expression of CC chemokine receptors (CCR) and CXC chemokine receptors (CXCR). Marrow stromal stem cells showed higher expression of CCR5 and CXCR4 chemokine receptors as compared to other types of chemokine receptors. Moreover, though the expression chemokine receptors generally decreased in most chemokine receptors with the cultivation of marrow stromal stem cells, CCR5 and CXCR4 chemokine receptors retained the higher lever of receptor expressions over prolonged periods. These results suggest that the ligands exhibiting specific binding to CCR5 or CXCR4 might be used to modify gene delivery systems for increased level of receptor-mediated gene delivery into stromal stem cells.