scholarly journals Osteogenic Matrix Cell Sheet Transplantation Enhances Early Tendon Graft to Bone Tunnel Healing in Rabbits

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Yusuke Inagaki ◽  
Kota Uematsu ◽  
Manabu Akahane ◽  
Yusuke Morita ◽  
Munehiro Ogawa ◽  
...  
Keyword(s):  
Early Stage ◽  
Cell Sheet ◽  
Collagen Type I ◽  
Bone Tunnel ◽  
Osteogenic Potential ◽  
Positive Staining ◽  
Type I ◽  
Micro Computed Tomography ◽  
Computed Tomography Images ◽  
Matrix Cell

The objective of this study was to determine whether osteogenic matrix cell sheets (OMCS) could induce bone formation around grafted tendons, thereby enhancing early stage tendon to bone tunnel healing in skeletally mature male Japanese white rabbits. First, the osteogenic potential of rabbit OMCS was evaluated. Then, the OMCS were transplanted into the interface between the grafted tendon and the bone tunnel created at the tibia. Histological assessments and biomechanical tensile testing were performed after 3 weeks. The rabbit OMCS showed high alkaline phosphatase (ALP) activity, positive staining of ALP, and osteogenic potential when transplanted subcutaneously with beta tricalcium phosphate disks. Newly formed bony walls and positive collagen type I staining were seen around the grafted tendon with OMCS transplantation, whereas such bony walls were thinner or less frequent without OMCS transplantation. Micro-computed tomography images showed significantly higher bone volume in the OMCS transplantation group. The pullout strength was significantly higher with OMCS (0.74±0.23 N/mm2) than without OMCS (0.58±0.15 N/mm2). These results show that OMCS enhance early tendon to bone tunnel healing. This method can be applied to cases requiring early tendon to bone tunnel healing after ligament reconstruction surgery.

scholarly journals Osteogenic Matrix Cell Sheets Facilitate Osteogenesis in Irradiated Rat Bone

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Yoshinobu Uchihara ◽  
Manabu Akahane ◽  
Takamasa Shimizu ◽  
Tomoyuki Ueha ◽  
Yusuke Morita ◽  
...  
Keyword(s):  
Treatment Option ◽  
Callus Formation ◽  
Cell Sheet ◽  
Bone Defects ◽  
Control Group ◽  
Micro Computed Tomography ◽  
Cell Sheets ◽  
Musculoskeletal Tumors ◽  
Computed Tomography Images ◽  
Matrix Cell

Reconstruction of large bone defects after resection of malignant musculoskeletal tumors is a significant challenge in orthopedic surgery. Extracorporeal autogenous irradiated bone grafting is a treatment option for bone reconstruction. However, nonunion often occurs because the osteogenic capacity is lost by irradiation. In the present study, we established an autogenous irradiated bone graft model in the rat femur to assess whether osteogenic matrix cell sheets improve osteogenesis of the irradiated bone. Osteogenic matrix cell sheets were prepared from bone marrow-derived stromal cells and co-transplanted with irradiated bone. X-ray images at 4 weeks after transplantation showed bridging callus formation around the irradiated bone. Micro-computed tomography images at 12 weeks postoperatively showed abundant callus formation in the whole circumference of the irradiated bone. Histology showed bone union between the irradiated bone and host femur. Mechanical testing showed that the failure force at the irradiated bone site was significantly higher than in the control group. Our study indicates that osteogenic matrix cell sheet transplantation might be a powerful method to facilitate osteogenesis in irradiated bones, which may become a treatment option for reconstruction of bone defects after resection of malignant musculoskeletal tumors.

scholarly journals Vancomycin-releasing cross-linked collagen sponges as wound dressings

2019 ◽  
Author(s):  
Jan Miroslav Hartinger ◽  
Peter Lukáč ◽  
Petr Mitáš ◽  
Mikuláš Mlček ◽  
Michaela Popková ◽  
...  
Keyword(s):  
Phosphate Buffer Solution ◽  
Collagen Type I ◽  
Wound Dressings ◽  
Homogeneous Distribution ◽  
Infrared Spectrometry ◽  
Type I ◽  
Micro Computed Tomography ◽  
Buffer Solution ◽  
Microbiological Testing

The study presents a novel vancomycin-releasing collagen wound dressing derived from Cyprinus carpio collagen type I cross-linked with carbodiimide which retarded the degradation rate and increased the stability of the sponge. Following lyophilization, the dressings were subjected to gamma sterilization. The structure was evaluated via scanning electron microscopy images, micro-computed tomography, and infrared spectrometry. The structural stability and vancomycin release properties were evaluated in a phosphate buffer solution. Microbiological testing and a rat model of a wound infected with methicillin-resistant Staphylococcus aureus (MRSA) were then employed to test the efficacy of the treatment of the infected wound. Following an initial mass loss due to the release of vancomycin, the sponges remained stable. After 7 days of exposure in phosphate buffered saline (37°C), 60% of the material remained with a preserved collagen secondary structure together with a high degree of open porosity (over 80%). The analysis of the release of the vancomycin revealed the homogeneous distribution of the antibiotic both across and between the sponges. The release of vancomycin was retarded as proved by in vitro testing and further confirmed by the animal model from which measurable concentrations were observed in blood samples 24 hours after the subcutaneous implantation of the sponge, which was more than observed following i. p. administration. The sponge was also highly effective in terms of reducing the number of colony-forming units in biopsies extracted from the infected wounds 4 days following the inoculation of the wounds with the MRSA solution.

scholarly journals The Osteogenic Potential of Human Nondifferentiated and Pre-differentiated Mesenchymal Stem Cells Combined with an Osteoconductive Scaffold – Early Stage Healing

2017 ◽  
Vol 60 (1) ◽  
pp. 12-18 ◽  
Author(s):  
Luboš Tuček ◽  
Zuzana Kočí ◽  
Kristýna Kárová ◽  
Helena Doležalová ◽  
Jakub Suchánek
Keyword(s):  
Stem Cells ◽  
Wistar Rats ◽  
Early Stage ◽  
Osteogenic Potential ◽  
Control Group ◽  
Positive Staining ◽  
Bone Scaffold ◽  
Synthetic Bone ◽  
Differentiated Cells ◽  
The Right

Despite the huge research into stem cells and their regenerative properties for bone healing, there are still unanswered questions including the recipient’s respond to the presence of the stem cells, the fate of stem cells inside the bone defect and the possible advantage in utilizing pre-differentiated cells. To address these problems, we used human multipotent mesenchymal stromal/stem cells (MSCs), GMP Grade, in a rat model of bone formation. In a “bioreactor concept” approach seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 2 × 106 MSCs, seven Wistar rats were implanted with 0.2 g of synthetic bone scaffold seeded with 1 × 106 predifferentiated osteoblasts and 1 × 106 pre-differentiated endothelial cells and 14 Wistar rats were implanted with 0.2 g of synthetic bone scaffold without seeded cells into an intramuscular pocket on the left side of their back. The right side of each rat was used as a control, and 0.2 g of synthetic bone scaffold was implanted into the intramuscular pocket alone. To see the early stage healing the samples were harvested 14 days after the implantation, MSCs were detected by positive DAPI and MTCO2 staining in 43% of all the samples implanted with MSCs, and no inflammation signs were present in any implanted animal. New vessels could be found in both groups implanted with MSCs, but not in the control group of animals. However, hematoxylin-eosin staining could not detect newly created bone within the implant in any of the groups. These results were in line with COLL1 staining, where we could detect positive staining only in three cases, all of which were implanted with un-differentiated MSCs. According to our findings, there were no benefits of using the pre-differentiated of MSC.

scholarly journals Laminin-521 Promotes Rat Bone Marrow Mesenchymal Stem Cell Sheet Formation on Light-Induced Cell Sheet Technology

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Zhiwei Jiang ◽  
Yue Xi ◽  
Kaichen Lai ◽  
Ying Wang ◽  
Huiming Wang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cell Sheet ◽  
Sheet Thickness ◽  
Collagen Type I ◽  
Type I ◽  
Cell Sheets ◽  
Cell Sheet Technology ◽  
Rat Bone

Rat bone marrow mesenchymal stem cell sheets (rBMSC sheets) are attractive for cell-based tissue engineering. However, methods of culturing rBMSC sheets are critically limited. In order to obtain intact rBMSC sheets, a light-induced cell sheet method was used in this study. TiO2 nanodot films were coated with (TL) or without (TN) laminin-521. We investigated the effects of laminin-521 on rBMSCs during cell sheet culturing. The fabricated rBMSC sheets were subsequently assessed to study cell sheet viability, reattachment ability, cell sheet thickness, collagen type I deposition, and multilineage potential. The results showed that laminin-521 could promote the formation of rBMSC sheets with good viability under hyperconfluent conditions. Cell sheet thickness increased from an initial 26.7 ± 1.5 μm (day 5) up to 47.7 ± 3.0 μm (day 10). Moreover, rBMSC sheets maintained their potential of osteogenic, adipogenic, and chondrogenic differentiation. This study provides a new strategy to obtain rBMSC sheets using light-induced cell sheet technology.

Gene Expressions Induced by Calcium Phosphate Ceramics after Implantation into the Muscle of Rat

2005 ◽  
Vol 475-479 ◽  
pp. 2363-2366
Author(s):  
Jin Rui Xu ◽  
Hong Song Fan ◽  
Yan Fei Tan ◽  
Xing Dong Zhang
Keyword(s):  
Calcium Phosphate ◽  
Collagen Type ◽  
Early Stage ◽  
Collagen Type I ◽  
Type I ◽  
Rt Pcr ◽  
Gene Expressions ◽  
Porous Hydroxyapatite ◽  
Calcium Phosphate Ceramics

The osteoinductivity of calcium phosphate ceramics has been studied extensively, but the mechanism is still unclear and few reports about the molecular mechanism in the osteoinductive process. In this study the osteoblast related gene expressions induced by biomaterials were investigated by isolating the RNA from the tissue grown in porous hydroxyapatite/tricalcium phosphate (HA/TCP) ceramics implanted in rat femur muscle on day 7, 15, 30, 60, 90,120, and analyzed by RT-PCR technique. RNA extracted from muscle without implant was used as control at the same time. The results showed that osteopontin and osteocalcin genes, the important osteoblastic markers, expressed in early stage, on day 7 after implantation, and were detected at any period. Collagen type I gene expressed on day 60, 90 and 120. It revealed that osteoblast differentiation occurred very early before collagen type I expression after implanting HA/TCP ceramics in vivo.

scholarly journals Studies on thePinctada fucataBMP-2 Gene: Structural Similarity and Functional Conservation of Its Osteogenic Potential within the Animal Kingdom

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Akiko Takami ◽  
Hirotaka Kato ◽  
Ryousuke Takagi ◽  
Tomoyuki Miyashita
Keyword(s):  
Structural Similarity ◽  
Collagen Type I ◽  
Osteoblastic Differentiation ◽  
Pinctada Fucata ◽  
Osteogenic Potential ◽  
Type I ◽  
Bioactive Substances ◽  
Functional Conservation ◽  
Morphogenetic Protein ◽  
Synthetic Polypeptide

Bone morphogenetic protein (BMP)-2 plays an important role in morphogenesis in both vertebrates and invertebrates. BMP-2 is one of the most powerful bioactive substances known to induce the osteogenic differentiation of mesenchymal cells. We examined the structural and functional conservation ofPinctada fucataBMP-2 in inducing osteogenesis in the murine mesenchymal stem cells, C3H10T1/2. Exposure of C3H10T1/2 cells to the recombinant mature fragment ofPinctada fucataBMP-2 resulted in osteoblastic differentiation. The sequence, SVPKPCCVPTELSSL, within the C-terminal portion ofPinctada fucataBMP-2, is homologous to the knuckle epitope of human BMP-2. This synthetic polypeptide was able to induce differentiation of C3H10T1/2 along the osteoblastic lineage, as confirmed by an increase in alkaline phosphatase activity, and the accumulation of calcium, as determined by von Kossa staining. Furthermore, using immunohistochemical staining, we observed an increased expression of collagen type I, osteopontin, and osteocalcin, which are known markers of osteogenesis. These results show that BMP-2 is conserved, not only in terms of its homology at the amino acid sequence, but also in terms of driving the formation of hard tissues in vertebrates and invertebrates.

scholarly journals A simvastatin-releasing scaffold with periodontal ligament stem cell sheets for periodontal regeneration

2020 ◽  
Vol 18 ◽  
pp. 228080001990009
Author(s):  
Bingjiao Zhao ◽  
Jing Chen ◽  
Liru Zhao ◽  
Jiajia Deng ◽  
Qiang Li
Keyword(s):  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Cell Sheet ◽  
Periodontal Regeneration ◽  
Collagen Type I ◽  
Type I ◽  
Bovine Bone ◽  
Sustained Drug Release ◽  
Periodontal Ligament Stem Cells ◽  
Membrane Scaffold

Simvastatin (SIM) has been documented to induce the osteogenic differentiation of periodontal ligament stem cells (PDLSCs). To establish an efficient release system for periodontal regeneration, a polycaprolactone (PCL) membrane scaffold containing SIM was electrospun and evaluated. The obtained PCL–SIM membrane scaffold showed sustained release up to 28 days, without deleterious effect on proliferation of PDLSCs on the scaffolds. PDLSCs were seeded onto scaffolds and their osteogenic differentiation was evaluated. After 21 days, expressions of collagen type I, alkaline phosphatase and bone sialoprotein genes were significantly upregulated and mineralized matrix formation was increased on the PCL–SIM scaffolds compared with the PCL scaffolds. In a heterotopic periodontal regeneration model, a cell sheet–scaffold construct was assembled by placement of multilayers of PDLSC sheets on PCL or PCL–SIM scaffolds, and these were then placed between dentin and ceramic bovine bone for subcutaneous implantation in athymic mice. After 8 weeks, the PCL–SIM membrane showed formation of significantly more ectopic cementum-like mineral on the dentin surface. These findings demonstrated that the PCL–SIM membrane scaffold promotes cementum-like tissue formation by sustained drug release, suggesting the feasibility of its therapeutic use with PDLSC sheets to improve periodontal regeneration.

scholarly journals Evaluation of candidate markers for the peritubular myoid cell lineage in the developing mouse testis

Reproduction ◽  
2005 ◽  
Vol 130 (4) ◽  
pp. 509-516 ◽  
Author(s):  
Angela Jeanes ◽  
Dagmar Wilhelm ◽  
Megan J Wilson ◽  
Josephine Bowles ◽  
Peter J McClive ◽  
...  
Keyword(s):  
Early Stage ◽  
Cell Lineage ◽  
Collagen Type I ◽  
Marker Genes ◽  
Type I ◽  
Myoid Cell ◽  
Fetal Testis ◽  
And Function ◽  
Peritubular Myoid Cell

Despite the importance of peritubular myoid (PM) cells in the histogenesis of the fetal testis, understanding the origin and function of these cells has been hampered by the lack of suitable markers. The current study was aimed at identifying molecular markers for PM cells during the early stages of testis development in the mouse embryo. Expression of candidate marker genes was tested by section in situ hybridisation, in some instances followed by immunofluorescent detection of protein products. Collagen type-I, inhibinβA, caldesmon 1 and tropomyosin 1 were found to be expressed by early-stage PM cells. These markers were also expressed in subsets of interstitial cells, most likely reflecting their common embryological provenance from migrating mesonephric cells. Although not strictly specific for PM cells, these markers are likely to be useful in studying the biology of early PM cells in the fetal testis.

Thermally Reversible Hydrogel Sheets for Adult Stem Cell Culture

2010 ◽  
Author(s):  
Sreedhar Thirumala ◽  
Ram V. Devireddy
Keyword(s):  
Stem Cell ◽  
Cell Sheet ◽  
Adult Stem Cell ◽  
Collagen Type I ◽  
Type I ◽  
Non Invasive ◽  
Bovine Collagen ◽  
Invasive Method ◽  
Water Salt ◽  
Coated Surface

A simple non-invasive method of cell retrieval using Methylcellulose (MC) coated Tissue Culture Poly-Styrone (TCPS) dishes was developed to allow the creation of single and multilayered adult stem cell sheet constructs while preserving the cell-cell and the cell–extracellular matrices. The optimal combination of MC-water-salt was found to be 12 to 14% of MC (mol. wt. of 15,000) in water with 0.5x PBS (∼150 mOsm). This solution exhibited a gel formation temperature of ∼32°C. The addition (evenly spread) of 200μl of 2mg/ml bovine collagen type -I (pH adjusted to 7.5) over the MC coated surface at 37°C, significantly improved the adult stem cell (ASC) adhesion and proliferation on the hydrogel system.

scholarly journals Surface Modification of Porous Titanium with Microarc Oxidation and Its Effects on Osteogenesis ActivityIn Vitro

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Qi Wang ◽  
Mengqi Cheng ◽  
Guo He ◽  
Xianlong Zhang
Keyword(s):  
De Novo ◽  
Early Stage ◽  
Microarc Oxidation ◽  
Porous Metal ◽  
Collagen Type I ◽  
Porous Titanium ◽  
Type I ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

Microarc oxidation (MAO) is a method about surface treatment that can provide nanoporous pits and thick oxide layers. A kind of porous metal-entangled titanium (Ti) wire material was treated with MAO process, resulting in a homogeneous rough TiO2layer, which helped facilitate MG-63 cell growth, cell viability, early cell differentiation, and cell mineralizationin vitro. In addition, the MAO-treated Ti surfaces could promote the proliferation of MG-63 cells without sacrificing differentiationin vitro, which would benefitde novobone formation around MAO-treated titanium at the early stage. The transcription levels of the extracellular matrix genes of osterix (OSX), collagen type I (Col I), bone sialoprotein (BSP), alkaline phosphatase (ALP), osteocalcin (OC) and osteopontin (OPN) and their protein expression levels were measured, suggesting that the cocultured cells with MAO titanium maintained the osteoblastic phenotype and that the MAO-treated titanium surface greatly stimulated osteoblast cell proliferation and differentiation compared to the untreated titanium. In conclusion, MAO technique can improve the surface of titanium and can contribute to the osseointegration process.

Sign in / Sign up

Export Citation Format

Share Document