scholarly journals Quantitative assessment of forward and backward second harmonic three dimensional images of collagen Type I matrix remodeling in a stimulated cellular environment

2012 ◽  
Vol 180 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Thomas Abraham ◽  
Damian Kayra ◽  
Bruce McManus ◽  
Alex Scott
Keyword(s):  
Quantitative Assessment ◽  
Collagen Type ◽  
Second Harmonic ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Type I ◽  
Matrix Remodeling ◽  
Cellular Environment ◽  
Three Dimensional Images

scholarly journals Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis

2016 ◽  
Vol 11 ◽  
pp. BMI.S38439 ◽  
Author(s):  
Federica Genovese ◽  
Zsolt S. Kàrpàti ◽  
Signe H. Nielsen ◽  
Morten A. Karsdal
Keyword(s):  
Extracellular Matrix ◽  
Collagen Type ◽  
Interstitial Fibrosis ◽  
Ex Vivo ◽  
Collagen Type I ◽  
Extracellular Matrix Remodeling ◽  
Type I ◽  
Matrix Remodeling ◽  
Renal Interstitial Fibrosis ◽  
Ex Vivo Model

The aim of this study was to set up an ex vivo model for renal interstitial fibrosis in order to investigate the extracellular matrix (ECM) turnover profile in the fibrotic kidney. We induced kidney fibrosis in fourteen 12-week-old male Sprague Dawley rats by unilateral ureteral obstruction (UUO) surgery of the right ureter. The left kidney (contralateral) was used as internal control. Six rats were sham operated and used as the control group. Rats were terminated two weeks after the surgery; the kidneys were excised and precision-cut kidney slices (PCKSs) were cultured for five days in serum-free medium. Markers of collagen type I formation (P1NP), collagen type I and III degradation (C1M and C3M), and α-smooth muscle actin (αSMA) were measured in the PCKS supernatants by enzyme-linked immunosorbent assay. P1NP, C1M, C3M, and α-SMA were increased up to 2- to 13-fold in supernatants of tissue slices from the UUO-ligated kidneys compared with the contralateral kidneys ( P < 0.001) and with the kidneys of sham-operated animals ( P < 0.0001). The markers could also reflect the level of fibrosis in different animals. The UUO PCKS ex vivo model provides a valuable translational tool for investigating the extracellular matrix remodeling associated with renal interstitial fibrosis.

Collagen Expression in Tissue Engineered Cartilage of Aged Human Articular Chondrocytes in a Rotating Bioreactor

2003 ◽  
Vol 26 (4) ◽  
pp. 319-330 ◽  
Author(s):  
S. Marlovits ◽  
B. Tichy ◽  
M. Truppe ◽  
D. Gruber ◽  
W. Schlegel
Keyword(s):  
Electron Microscopy ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Human Articular Chondrocytes ◽  
Type I ◽  
Transmission Electron ◽  
Low Shear Stress ◽  
Engineered Cartilage

This study describes the culture and three-dimensional assembly of aged human articular chondrocytes under controlled oxygenation and low shear stress in a rotating-wall vessel. Chondrocytes cultured in monolayer were released and placed without any scaffold as a single cell suspension in a rotating bioreactor for 12 weeks. Samples were analyzed with immunohistochemistry, molecular biology and electron microscopy. During serial monolayer cultures chondrocytes dedifferentiated to a “fibroblast-like” structure and produced predominantly collagen type I. When these dedifferentiated cells were transferred to the rotating bioreactor, the cells showed a spontaneous aggregation and formation of solid tissue during the culture time. Expression of collagen type II and other components critical for the extracellular cartilage matrix could be detected. Transmission electron microscopy revealed a fine network of randomly distributed collagen fibrils. This rotating bioreactor proves to be a useful tool for providing an environment that enables dedifferentiated chondrocytes to redifferentiate and produce a cartilage-specific extracellular matrix.

Mesenchymal stem cells from adipose tissue which have been differentiated into chondrocytes in three-dimensional culture express lubricin

2011 ◽  
Vol 236 (11) ◽  
pp. 1333-1341 ◽  
Author(s):  
Giuseppe Musumeci ◽  
Debora Lo Furno ◽  
Carla Loreto ◽  
Rosario Giuffrida ◽  
Silvia Caggia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Collagen Type ◽  
Three Dimensional ◽  
3D Culture ◽  
Collagen Type I ◽  
Type I ◽  
Human Mscs ◽  
Alternative Source

The present study focused on the isolation, cultivation and characterization of human mesenchymal stem cells (MSCs) from adipose tissue and on their differentiation into chondrocytes through the NH ChondroDiff medium. The main aim was to investigate some markers of biomechanical quality of cartilage, such as lubricin, and collagen type I and II. Little is known, in fact, about the ability of chondrocytes from human MSCs of adipose tissue to generate lubricin in three-dimensional (3D) culture. Lubricin, a 227.5-kDa mucinous glycoprotein, is known to play an important role in articular joint physiology, and the loss of accumulation of lubricin is thought to play a role in the pathology of osteoarthritis. Adipose tissue is an alternative source for the isolation of multipotent MSCs, which allows them to be obtained by a less invasive method and in larger quantities than from other sources. These cells can be isolated from cosmetic liposuctions in large numbers and easily grown under standard tissue culture conditions. 3D chondrocytes were assessed by histology (hematoxylin and eosin) and histochemistry (Alcian blue and Safranin-O/fast green staining). Collagen type I, II and lubricin expression was determined through immunohistochemistry and Western blot. The results showed that, compared with control cartilage and monolayer chondrocytes showing just collagen type I, chondrocytes from MSCs (CD44-, CD90- and CD105- positive; CD45-, CD14- and CD34-negative) of adipose tissue grown in nodules were able to express lubricin, and collagen type I and II, indicative of hyaline cartilage formation. Based on the function of lubricin in the joint cavity and disease and as a potential therapeutic agent, our results suggest that MSCs from adipose tissue are a promising cell source for tissue engineering of cartilage. Our results suggest that chondrocyte nodules producing lubricin could be a novel biotherapeutic approach for the treatment of cartilage abnormalities.

scholarly journals Preliminary Study of In Vitro Three-Dimensional Skin Model Using an Ovine Collagen Type I Sponge Seeded with Co-Culture Skin Cells: Submerged versus Air-Liquid Interface Conditions

Polymers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 2784
Author(s):  
Mh Busra Fauzi ◽  
Zahra Rashidbenam ◽  
Aminuddin Bin Saim ◽  
Ruszymah Binti Hj Idrus
Keyword(s):  
Collagen Type ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Liquid Interface ◽  
Type I ◽  
Skin Model ◽  
Skin Cells ◽  
Air Liquid Interface ◽  
In Vitro Skin Model

Three-dimensional (3D) in vitro skin models have been widely used for cosmeceutical and pharmaceutical applications aiming to reduce animal use in experiment. This study investigate capability of ovine tendon collagen type I (OTC-I) sponge suitable platform for a 3D in vitro skin model using co-cultured skin cells (CC) containing human epidermal keratinocytes (HEK) and human dermal fibroblasts (HDF) under submerged (SM) and air-liquid interface (ALI) conditions. Briefly, the extracted OTC-I was freeze-dried and crosslinked with genipin (OTC-I_GNP) and carbodiimide (OTC-I_EDC). The gross appearance, physico-chemical characteristics, biocompatibility and growth profile of seeded skin cells were assessed. The light brown and white appearance for the OTC-I_GNP scaffold and other groups were observed, respectively. The OTC-I_GNP scaffold demonstrated the highest swelling ratio (~1885%) and water uptake (94.96 ± 0.14%). The Fourier transformation infrared demonstrated amide A, B and I, II and III which represent collagen type I. The microstructure of all fabricated sponges presented a similar surface roughness with the presence of visible collagen fibers and a heterogenous porous structure. The OTC-I_EDC scaffold was more toxic and showed the lowest cell attachment and proliferation as compared to other groups. The micrographic evaluation revealed that CC potentially formed the epidermal- and dermal-like layers in both SM and ALI that prominently observed with OTC-I_GNP compared to others. In conclusion, these results suggest that OTC_GNP could be used as a 3D in vitro skin model under ALI microenvironment.

scholarly journals Therapeutic Effects of Acupuncture through Enhancement of Functional Angiogenesis and Granulogenesis in Rat Wound Healing

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Sang In Park ◽  
Yun-Young Sunwoo ◽  
Yu Jin Jung ◽  
Woo Chul Chang ◽  
Moon-Seo Park ◽  
...  
Keyword(s):  
Wound Healing ◽  
Collagen Type ◽  
Interleukin 1 ◽  
Treated Group ◽  
Collagen Type I ◽  
Control Group ◽  
Therapeutic Effects ◽  
Type I ◽  
Matrix Remodeling ◽  
Cd 31

Acupuncture regulates inflammation process and growth factors by increasing blood circulation in affected areas. In this study, we examined whether acupuncture has an effect on wound healing in injured rat. Rats were assigned randomly into two groups: control group and acupuncture group. Acupuncture treatment was carried out at 8 sites around the wounded area. We analyzed the wound area, inflammatory cytokines, proliferation of resident cells, and angiogenesis and induction of extracelluar matrix remodeling. At 7 days after-wounding the wound size in acupuncture-treat group was decreased more significantly compared to control group. In addition, the protein levels of proinflammatory cytokines such as tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were significantly decreased compared to the control at 2 and 7 days post-wounding. Also, we analyzed newly generated cells by performing immunostaining for PCNA and using several phenotype markers such as CD-31,α-SMA, and collagen type I. In acupuncture-treated group, PCNA-positive cell was increased and PCNA labeled CD-31-positive vessels,α-SMA- and collagen type I-positive fibroblastic cells, were increased compared to the control group at 7 days post-wounding. These results suggest that acupuncture may improve wound healing through decreasing pro-inflammatory response, increasing cell proliferation and angiogenesis, and inducing extracellular matrix remodeling.

Biodegradable Nano-Material Composites for Use in an Inkjet Printing System

2006 ◽  
Vol 921 ◽  
Author(s):  
Nicole H Levi ◽  
John B. McGuirt ◽  
Faith M. Coldren ◽  
David L. Carroll
Keyword(s):  
Inkjet Printing ◽  
Collagen Type ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Type I ◽  
Printing System ◽  
Walled Carbon Nanotubes ◽  
Thermal Inkjet Printing ◽  
Compatibility Of Materials ◽  
Insight Into

AbstractBiomaterials for development of resorbable, three-dimensional tissue scaffolds have been used in a modified thermal inkjet printing system to explore compatibility of materials, solvents and the printing system. The polymers included collagen (type I), sodium alginate, fibronectin, poly-lactic co-glycolic acid (PLGA), polyethylene glycol (PEG), and tetraglycol were tested. Single-walled carbon nanotubes were combined with the biopolymers to determine which systems in which they would blend well, and be able to print. Uncovering which biopolymers may be printed together offers insight into development of materials which most closely match the properties of biological tissue.

scholarly journals Stokes polarimetry-based second harmonic generation microscopy for collagen and skeletal muscle fiber characterization

2020 ◽  
Author(s):  
Nirmal Mazumder ◽  
Fu-Jen Kao
Keyword(s):  
Skeletal Muscle ◽  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Muscle Fiber ◽  
Collagen Type ◽  
Second Harmonic ◽  
Collagen Type I ◽  
Skeletal Muscle Fiber ◽  
Type I ◽  
Stokes Vector

Abstract The complete polarization state of second harmonic (SH) light was measured and characterized by collagen type I and skeletal muscle fiber using a Stokes vector-based SHG microscope. The polarization states of the SH signal are analyzed in a pixel-by-pixel manner and displayed through two dimensional (2D) Stokes vector images. Various polarization parameters are reconstructed using Stokes values to quantify the polarization properties of SH light. Also, the measurements are extended for different input polarization states to investigate the molecular structure of second harmonic generation (SHG) active molecules such as collagen type I and myosin.

Effects of Mechanical Strain on Osteoblastic Precursor Cells in a Three-Dimensional Scaffold

2007 ◽  
Vol 330-332 ◽  
pp. 1181-1184
Author(s):  
Zhi He Zhao ◽  
Jun Wang ◽  
Yu Bo Fan ◽  
Song Jiao Luo ◽  
Ling Yong Jiang
Keyword(s):  
Collagen Type ◽  
Mechanical Strain ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Precursor Cells ◽  
Dynamic Strain ◽  
Type I ◽  
Pcr Assay ◽  
Periodontal Tissues ◽  
Strain Magnitude

It was well recognized that mechanical strain plays a crucial role in periodontal tissues remodeling. The aim of this study was to investigate the effect of mechanical strain on osteoblastic precursor cells in a collagen type I gel scaffold. Rat MSCs were isolated and cultured according to the established method. Cells were induced with osteogenic medium, then seeded in a collagen type I gel and mechanically stretched by application of cyclic biaxial strain 24h later. Strain cycle was set to 1 cycle/min (0.017Hz), and strain magnitude was set to 2%, 5%, 7% elongation. Cells were collected in 0h, 3h, 6h, 9h, 12h, 24h and 48h respectively. ODF and ICAM-1 mRNA were analyzed by RT-PCR assay. The results shown that 2-7% elongation strain, either dynamic or static, inhibited ICAM-1and ODF expression of osteoblastic precursors, and the effects were relative tightly to strain magnitude. The inhibition effects of dynamic strain loading group exceeded the corresponding static strain. This work suggested that appropriate mechanical strech may suppress differentiation of osteoclasts through inhibiting expression of ICAM-1 and ODF. Application of mechanical stress might have a beneficial effect on quantity of generated bone tissue and might be a important factor in tissue engineering of periodontal tissues.

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