Cell sheet transplantation of cultured mesenchymal stem cells enhances bone formation in a rat nonunion model

Bone ◽  
2010 ◽  
Vol 46 (2) ◽  
pp. 418-424 ◽  
Author(s):  
Akifumi Nakamura ◽  
Manabu Akahane ◽  
Hideki Shigematsu ◽  
Mika Tadokoro ◽  
Yusuke Morita ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Cell Sheet

scholarly journals Biomimetic vascularized adipose-derived mesenchymal stem cells bone-periosteum graft enhances angiogenesis and osteogenesis in a rabbit spine fusion model

2021 ◽  
Author(s):  
Tsai-Sheng Fu ◽  
Wei-Chuan Chen ◽  
Ying-Chih Wang ◽  
Chia-Wei Chang ◽  
Tung-Yi Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Blood Vessels ◽  
Critical Role ◽  
Cell Sheet ◽  
Biomechanical Testing ◽  
Bone Grafts ◽  
Vascularized Bone

Abstract BackgroundSeveral artificial bone grafts have been developed for bone reconstruction but fail to achieve anticipated osteogenesis due to their insufficient neovascularization capacity. Besides, periosteum plays an essential role in the neovascularization process in bone formation and healing. The aim of this study was to develop a cell-based biomimetic vascularized bone-periosteum construct (VBPC) to provide better neovascularization for osteogenesis and bone regeneration.MethodsTwenty-four male New Zealand white rabbits were divided into four groups according to the experimental materials. We first cultured adipose-derived mesenchymal stem cells (AMSCs) and seeded them evenly in the collagen/chitosan sheet to form an AMSCs-sheet-engineered periosteum. Simultaneously, the AMSCs were seeded onto alginate scaffolds and were cultured to differentiate to endothelial-like cells to form vascularized bone constructs (VBC). The success of endothelial differentiation was confirmed by real-time polymerase chain reaction and immunofluorescence staining analysis. The AMSCs-sheet-engineered periosteum was wrapped onto VBC to create biomimetic VBPC, which was then implanted in bilateral L4-5 intertransverse space of rabbit. The acellular alginate-sheet construct, VBC, and non-vascularized AMSCs-alginate-periosteum construct were used as controls. At 12 weeks after implantation, the bone-forming capacities of the constructs were determined by computed tomography, biomechanical testing, histology, and immunohistochemistry staining analyses.ResultsTwelve weeks after implantation, the VBPC group significantly increased new bone formation volume than the control groups. Biomechanical testing demonstrated a higher torque strength in the VBPC group, and suggested that the cell sheet played a critical role for mechanical support. Notably, the hematoxylin and eosin, Masson’s trichrome, and immunohistochemistry stained histologic results revealed that the VBPC group promoted the formation of blood vessels and new bones in the L4-5 intertransverse fusion areas.ConclusionsThe tissue-engineered biomimetic VBPC showed great capability in promoting angiogenesis and osteogenesis in vivo. The VBPC may overcome the deficits of traditional bone grafts. These findings suggest a novel approach to improve the timely formation of blood vessels from bone substitutes and provide an ideal source for bone regeneration.

scholarly journals A novel gelatin/chitooligosaccharide/demineralized bone matrix composite scaffold and periosteum-derived mesenchymal stem cells for bone tissue engineering

2021 ◽  
Vol 25 (1) ◽  
Author(s):  
Thakoon Thitiset ◽  
Siriporn Damrongsakkul ◽  
Supansa Yodmuang ◽  
Wilairat Leeanansaksiri ◽  
Jirun Apinun ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Alp Activity

Abstract Background A novel biodegradable scaffold including gelatin (G), chitooligosaccharide (COS), and demineralized bone matrix (DBM) could play a significant part in bone tissue engineering. The present study aimed to investigate the biological characteristics of composite scaffolds in combination of G, COS, and DBM for in vitro cell culture and in vivo animal bioassays. Methods Three-dimensional scaffolds from the mixture of G, COS, and DBM were fabricated into 3 groups, namely, G, GC, and GCD using a lyophilization technique. The scaffolds were cultured with mesenchymal stem cells (MSCs) for 4 weeks to determine biological responses such as cell attachment and cell proliferation, alkaline phosphatase (ALP) activity, calcium deposition, cell morphology, and cell surface elemental composition. For the in vivo bioassay, G, GC, and GCD, acellular scaffolds were implanted subcutaneously in 8-week-old male Wistar rats for 4 weeks and 8 weeks. The explants were assessed for new bone formation using hematoxylin and eosin (H&E) staining and von Kossa staining. Results The MSCs could attach and proliferate on all three groups of scaffolds. Interestingly, the ALP activity of MSCs reached the greatest value on day 7 after cultured on the scaffolds, whereas the calcium assay displayed the highest level of calcium in MSCs on day 28. Furthermore, weight percentages of calcium and phosphorus on the surface of MSCs after cultivation on the GCD scaffolds increased when compared to those on other scaffolds. The scanning electron microscopy images showed that MSCs attached and proliferated on the scaffold surface thoroughly over the cultivation time. Mineral crystal aggregation was evident in GC and greatly in GCD scaffolds. H&E staining illustrated that G, GC, and GCD scaffolds displayed osteoid after 4 weeks of implantation and von Kossa staining confirmed the mineralization at 8 weeks in G, GC, and GCD scaffolds. Conclusion The MSCs cultured in GCD scaffolds revealed greater osteogenic differentiation than those cultured in G and GC scaffolds. Additionally, the G, GC, and GCD scaffolds could promote in vivo ectopic bone formation in rat model. The GCD scaffolds exhibited maximum osteoinductive capability compared with others and may be potentially used for bone regeneration.

scholarly journals Regeneration in Experimental Alveolar Bone Defect Using Human Umbilical Cord Mesenchymal Stem Cells

2021 ◽  
Vol 30 ◽  
pp. 096368972097539
Author(s):  
Akiko Toyota ◽  
Rei Shinagawa ◽  
Mikiko Mano ◽  
Kazuyuki Tokioka ◽  
Naoto Suda
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Cleft Lip ◽  
Human Umbilical Cord ◽  
Bone Surface ◽  
Bone Bridge ◽  
Bridge Formation ◽  
Alveolar Cleft ◽  
Umbilical Cords

Cleft lip and palate is a congenital disorder including cleft lip, and/or cleft palate, and/or alveolar cleft, with high incidence.The alveolar cleft causes morphological and functional abnormalities. To obtain bone bridge formation and continuous structure between alveolar clefts, surgical interventions are performed from infancy to childhood. However, desirable bone bridge formation is not obtained in many cases. Regenerative medicine using mesenchymal stem cells (MSCs) is expected to be a useful strategy to obtain sufficient bone bridge formation between alveolar clefts. In this study, we examined the effect of human umbilical cord-derived MSCs by transplantation into a rat experimental alveolar cleft model. Human umbilical cords were digested enzymatically and the isolated cells were collected (UC-EZ cells). Next, CD146-positive cells were enriched from UC-EZ cells by magnetic-activated cell sorting (UC-MACS cells). UC-EZ and UC-MACS cells showed MSC gene/protein expression, in vitro. Both cells had multipotency and could differentiate to osteogenic, chondrogenic, and adipogenic lineages under the differentiation-inducing media. However, UC-EZ cells lacked Sox2 expression and showed the lower ratio of MSCs than UC-MACS cells. Thus, UC-MACS cells were transplanted with hydroxyapatite and collagen (HA + Col) into alveolar cleft model to evaluate bone formation in vivo. The results of micro computed tomography and histological staining showed that UC-MACS cells with HA + Col induced more abundant bone formation between the experimental alveolar clefts than HA + Col implantation only. Cells immunopositive for osteopontin were accumulated along the bone surface and some of them were embedded in the bone. Cells immunopositive for human-specific mitochondria were aligned along the newly formed bone surface and in the new bone, suggesting that UC-MACS cells contributed to the bone bridge formation between alveolar clefts. These findings indicate that human umbilical cords are reliable bioresource and UC-MACS cells are useful for the alveolar cleft regeneration.

scholarly journals miR-16-2* Interferes with WNT5A to Regulate Osteogenesis of Mesenchymal Stem Cells

2018 ◽  
Vol 51 (3) ◽  
pp. 1087-1102 ◽  
Author(s):  
Lijun Duan ◽  
He Zhao ◽  
Yang Xiong ◽  
Xiangsheng Tang ◽  
Yongdong Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Target Genes ◽  
Fragility Fractures ◽  
Inhibitory Effect ◽  
Lymphocytic Leukemia ◽  
Qrt Pcr ◽  
Wnt Signal

Background/Aims: Osteoporosis is a bone metabolic disease characterized by a systemic impairment of bone mass, which results in increased propensity of fragility fractures. A reduction in the differentiation of MSCs into osteoblasts contributes to the impaired bone formation observed in osteoporosis. Mesenchymal stem cells (MSCs) are induced to differentiate into preosteoblasts, which are regulated by the signaling cascades initiated by the various signals, including miRNAs. miR-16-2* is a newly discovered miRNA that participates in diagnosis and prognosis of hepatocellular carcinoma, cervical cancer and chronic lymphocytic leukemia. However, the effect of miR-16-2* on the regulation of osteoblast differentiation and the mechanism responsible are still unclear. Here we discuss the contribution of miR-16-2* to osteoporosis, osteoblast differentiation and mineralization. Methods: The expression pattern of miR-16-2* during osteogenesis or in osteoporosis bone samples was validated by quantitative real-time PCR (qRT-PCR). The human bone marrow mesenchymal stem cells (hBMSCs) were induced to differentiate into osteoblasts by osteogenic induced medium containing dexamethasone, ascorbate-2-phosphat, beta-glycerophosphate and vitamin-D3. The target genes of miR-16-2* were predicted by TargetScan and PicTar. The mRNA and protein levels of osteogenic key markers were detected using qRT-PCR or western blot respectively. The WNT signal activity was analyzed by TOP/FOP reporter assay. Results: The expression of miR-16-2* in patient bone tissue with osteoporosis was negatively correlated with bone formation related genes. During osteoblast differentiation process, the expression of miR-16-2* was significantly decreased. Upregulation of miR-16-2* in hBMSCs impaired the osteogenic differentiation while the downregulation of miR-16-2* increased this process. Upregulation the expression of miR-16-2* could also block the WNT signal pathway by directly target WNT5A. Furthermore, knockdown of miR-16-2* could promote the activation of RUNX2, possibly by lifting the inhibitory effect of miR-16-2* on WNT pathway. Conclusion: Taken together, we report a novel biological role of miR-16-2* in osteogenesis through regulating WNT5A response for the first time. Our data support the potential utilization of miRNA-based therapies in regenerative medicine.

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