scholarly journals Small-Diameter Subchondral Drilling Improves DNA and Proteoglycan Content of the Cartilaginous Repair Tissue in a Large Animal Model of a Full-Thickness Chondral Defect

2020 ◽  
Vol 9 (6) ◽  
pp. 1903 ◽  
Author(s):  
Patrick Orth ◽  
Mona Eldracher ◽  
Magali Cucchiarini ◽  
Henning Madry
Keyword(s):  
Type I Collagen ◽  
Small Diameter ◽  
Large Animal Model ◽  
Large Animal ◽  
Type I ◽  
Type Ii ◽  
Chondral Defect ◽  
Repair Tissue ◽  
Proteoglycan Content ◽  
Adjacent Cartilage

This study quantified changes in the DNA content and extracellular matrix composition of both the cartilaginous repair tissue and the adjacent cartilage in a large animal model of a chondral defect treated by subchondral drilling. Content of DNA, proteoglycans, and Type II and Type I collagen, as well as their different ratios were assessed at 6 months in vivo after treatment of full-thickness cartilage defects in the femoral trochlea of adult sheep with six subchondral drill holes, each of either 1.0 mm or 1.8 mm in diameter by biochemical analyses of the repair tissue and the adjacent cartilage and compared with the original cartilage. Only subchondral drilling which were 1.0 mm in diameter significantly increased both DNA and proteoglycan content of the repair tissue compared to the original cartilage. DNA content correlated with the proteoglycan and Type II collagen content within the repair tissue. Significantly higher amounts of Type I collagen within the repair tissue and significantly increased DNA, proteoglycan, and Type I collagen content in the adjacent cartilage were identified. These translational data support the use of small-diameter bone-cutting devices for marrow stimulation. Signs of early degeneration were present within the cartilaginous repair tissue and the adjacent cartilage.

Short oxygen prebreathe periods reduce or prevent severe decompression sickness in a 70-kg swine saturation model

2009 ◽  
Vol 106 (4) ◽  
pp. 1459-1463 ◽  
Author(s):  
R. T. Mahon ◽  
H. M. Dainer ◽  
M. G. Gibellato ◽  
S. E. Soutiere
Keyword(s):  
Decompression Sickness ◽  
Inert Gas ◽  
Large Animal Model ◽  
Large Animal ◽  
Type I ◽  
Type Ii ◽  
Saturation Model ◽  
Hyperbaric Chamber ◽  
Improve Outcome ◽  
Rescue Procedures

Disabled submarine (DISSUB) survivors are expected to achieve saturation with inert gas. However, rescue procedures may not accommodate staged decompression, raising the potential for severe decompression sickness (DCS). Alternatives to standard recompression therapy are needed. It has been demonstrated in humans that isobaric oxygen “prebreathing” (OPB) can accelerate decompression in a DISSUB scenario. In-70 kg swine saturated at 2.82 atm absolute (ATA), 1 h of OPB eliminated death and reduced severe DCS. We hypothesized that even shorter periods (<1 h) of OPB before no-stop decompression from saturation at 2.82 ATA could reduce the incidence of DCS in a large animal model. Catheterized Yorkshire swine (68.8 ± 1.7 kg) in individual Plexiglas boxes within a large animal hyperbaric chamber were compressed to 2.82 ATA for 22 h. Following saturation and while still at depth, breathing gas was switched to >95% O2 for 45 min (OPB45), 15 min (OPB15), or 5 min (OPB05) of OPB, or no OPB (control). The chamber was then decompressed without stops (0.91 ATA/min). Observers then entered the chamber and recorded signs of DCS for 2 h. All OPB periods significantly reduced the risk of developing type II DCS. OPB45 eliminated severe DCS. Controls had a 2.5 times greater risk of developing type II DCS than OPB05 ( P = 0.016). OPB45 and OPB15 significantly reduced type I DCS compared with controls. These results support the potential of OPB as an alternative to staged decompression and that OPB could be expected to improve outcome in a DISSUB rescue scenario.

Histologic and Immunohistochemical Characteristics of Failed Articular Cartilage Resurfacing Procedures for Osteochondritis of the Knee

2007 ◽  
Vol 36 (2) ◽  
pp. 360-368 ◽  
Author(s):  
Robert F. LaPrade ◽  
Laura S. Bursch ◽  
Erik J. Olson ◽  
Vojtech Havlas ◽  
Cathy S. Carlson
Keyword(s):  
Articular Cartilage ◽  
Autologous Chondrocyte Implantation ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Repair Tissue ◽  
Tissue Area ◽  
Chondrocyte Implantation ◽  
Autologous Chondrocyte

Background The histologic appearance of the repair tissue after articular cartilage resurfacing procedures in humans is not well documented. Hypothesis The histologic and immunohistochemical appearance of the repair tissues in failed articular cartilage resurfacing procedures will be similar, regardless of the procedure that was done, and will not resemble normal articular cartilage. Study Design Case series; Level of evidence, 4. Methods Graft tissue from 10 patients who underwent an autologous chondrocyte implantation (n = 6), microfracture (n = 3), or periosteal transplantation (n = 1) procedure to treat symptomatic osteochondritis dissecans of the medial femoral condyle was processed for histologic examination after failure of the articular cartilage resurfacing procedure. Serial sections from all slabs were stained with hematoxylin and eosin and toluidine blue and were immunostained using antibodies directed against types I, II, and X collagen. Results Specimens from all 3 types of repair procedures were composed primarily of fibrous connective tissue and fibrocartilage. None of the sections stained positively for type X collagen. All 10 cases stained positively for type I collagen (range, 7%–97% of tissue area). Staining for type II collagen was positive in 4 of 6 autologous chondrocyte implantation cases, 3 of 3 microfracture cases, and the periosteal transplant case (range, 2%–65% of tissue area). In 8 of 10 cases, the percentage of the section area exhibiting positive staining for type I collagen was higher than for type II collagen (6 of 6 autologous chondrocyte implantation; 1 of 3 microfracture; 1 periosteal transplant). Conclusion The histologic appearance of the repair tissue of 3 different failed articular cartilage resurfacing procedures was similar and did not resemble normal articular cartilage.

scholarly journals P1205ESTABLISHMENT OF HYPOCHLORITE-INDUCED PORCINE MODEL OF PERITONEAL FIBROSIS

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Chun-Yuan Chao ◽  
Chin-Ho Wu ◽  
Yi-Ting Chen ◽  
SHUEI-LIONG LIN ◽  
Pei-Shiue Tsai
Keyword(s):  
Peritoneal Dialysis ◽  
Type I Collagen ◽  
Porcine Model ◽  
Large Animal Model ◽  
Large Animal ◽  
Type I ◽  
Peritoneal Fibrosis ◽  
Life Saving ◽  
Peritoneal Mesothelium ◽  
High Degree

Abstract Background and Aims Patients with kidney failure rely on life-saving peritoneal dialysis to facilitate waste exchange and maintain homeostasis of physical conditions. However, peritoneal dialysis often results in peritoneal fibrosis and organ adhesion that subsequently compromise the efficiency of peritoneal dialysis and normal functions of visceral organs. Despite rodent models provide clues on the pathogenesis of peritoneal fibrosis, no current large animal model which shares high degree of physiological and anatomical similarities to human is available, limiting their applications on the evaluation of pre-clinical therapeutic efficacy. Method In this study, we established for the first time, porcine model of peritoneal fibrosis by the use of a bleach-like chemical, sodium hypochlorite. We demonstrated that intraperitoneal injection of 30ml/kg B.W., 0.1%-0.2% (0.1mM-0.2mM) hypochlorite induced peritoneal fibrosis and visceral organ adhesions in 5-week-old piglets. Results A dose- and time-dependent severity of peritoneal fibrosis characterized by mesothelium fragmentation, αSMA+ myofibroblasts accumulation, organ surface thickening and type I collagen deposition were observed. We also demonstrated that hypochlorite-induced overexpression of IL1β, CX3CL1 and TGFβ on the peritoneal mesothelium mimicked the mechanism of peritoneal dialysis-induced peritoneal fibrosis in human patients. Conclusion This pig model could not only be used as a platform for studying fibrosis/scar formation, but can also be used to evaluate the efficacy of potential candidates on the prevention (e.g. compounds) and treatments (e.g. stem cells) for regenerative medicine.

scholarly journals Intracellular and Extracellular Markers of Lethality in Osteogenesis Imperfecta: A Quantitative Proteomic Approach

2021 ◽  
Vol 22 (1) ◽  
pp. 429
Author(s):  
Luca Bini ◽  
Domitille Schvartz ◽  
Chiara Carnemolla ◽  
Roberta Besio ◽  
Nadia Garibaldi ◽  
...  
Keyword(s):  
Osteogenesis Imperfecta ◽  
Type I Collagen ◽  
Type I ◽  
Tandem Mass ◽  
Type Ii ◽  
Type Iii ◽  
Bioinformatic Tools ◽  
Proteomic Approach ◽  
Fibrillin 1 ◽  
Heritable Disorder

Osteogenesis imperfecta (OI) is a heritable disorder that mainly affects the skeleton. The inheritance is mostly autosomal dominant and associated to mutations in one of the two genes, COL1A1 and COL1A2, encoding for the type I collagen α chains. According to more than 1500 described mutation sites and to outcome spanning from very mild cases to perinatal-lethality, OI is characterized by a wide genotype/phenotype heterogeneity. In order to identify common affected molecular-pathways and disease biomarkers in OI probands with different mutations and lethal or surviving phenotypes, primary fibroblasts from dominant OI patients, carrying COL1A1 or COL1A2 defects, were investigated by applying a Tandem Mass Tag labeling-Liquid Chromatography-Tandem Mass Spectrometry (TMT LC-MS/MS) proteomics approach and bioinformatic tools for comparative protein-abundance profiling. While no difference in α1 or α2 abundance was detected among lethal (type II) and not-lethal (type III) OI patients, 17 proteins, with key effects on matrix structure and organization, cell signaling, and cell and tissue development and differentiation, were significantly different between type II and type III OI patients. Among them, some non–collagenous extracellular matrix (ECM) proteins (e.g., decorin and fibrillin-1) and proteins modulating cytoskeleton (e.g., nestin and palladin) directly correlate to the severity of the disease. Their defective presence may define proband-failure in balancing aberrances related to mutant collagen.

Type VI collagen expression is upregulated in the early events of chondrocyte differentiation

Development ◽  
1993 ◽  
Vol 117 (1) ◽  
pp. 245-251
Author(s):  
R. Quarto ◽  
B. Dozin ◽  
P. Bonaldo ◽  
R. Cancedda ◽  
A. Colombatti
Keyword(s):  
Suspension Culture ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Type Vi Collagen ◽  
Full Spectrum ◽  
Collagen Expression ◽  
Hypertrophic Chondrocyte

Dedifferentiated chondrocytes cultured adherent to the substratum proliferate and synthesize large amounts of type I collagen but when transferred to suspension culture they decrease proliferation, resume the chondrogenic phenotype and the synthesis of type II collagen, and continue their maturation to hypertrophic chondrocyte (Castagnola et al., 1986, J. Cell Biol. 102, 2310–2317). In this report, we describe the developmentally regulated expression of type VI collagen in vitro in differentiating avian chondrocytes. Type VI collagen mRNA is barely detectable in dedifferentiated chondrocytes as long as the attachment to the substratum is maintained, but increases very rapidly upon passage of the cells into suspension culture reaching a peak after 48 hours and declining after 5–6 days of suspension culture. The first evidence of a rise in the mRNA steady-state levels is obtained already at 6 hours for the alpha 3(VI) chain. Immunoprecipitation of metabolically labeled cells with type VI collagen antibodies reveals that the early mRNA rise is paralleled by an increased secretion of type VI collagen in cell media. Induction of type VI collagen is not the consequence of trypsin treatment of dedifferentiated cells since exposure to the actin-disrupting drug cytochalasin or detachment of the cells by mechanical procedures has similar effects. In 13-day-old chicken embryo tibiae, where the full spectrum of the chondrogenic differentiation process is represented, expression of type VI collagen is restricted to the articular cartilage where chondrocytes developmental stage is comparable to stage I (high levels of type II collagen expression).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Role of collagenase in mediating in vitro alveolar epithelial wound repair

1999 ◽  
Vol 112 (2) ◽  
pp. 243-252
Author(s):  
E. Planus ◽  
S. Galiacy ◽  
M. Matthay ◽  
V. Laurent ◽  
J. Gavrilovic ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Type I Collagen ◽  
Alveolar Epithelial Cell ◽  
Cell Monolayer ◽  
Type I ◽  
Type Ii ◽  
Alveolar Epithelial

Type II pneumocytes are essential for repair of the injured alveolar epithelium. The effect of two MMP collagenases, MMP-1 and MMP-13 on alveolar epithelial repair was studied in vitro. The A549 alveolar epithelial cell line and primary rat alveolar epithelial cell cultures were used. Cell adhesion and cell migration were measured with and without exogenous MMP-1. Wound healing of a cell monolayer of rat alveolar epithelial cell after a mechanical injury was evaluated by time lapse video analysis. Cell adhesion on type I collagen, as well as cytoskeleton stiffness, was decreased in the presence of exogenous collagenases. A similar decrease was observed when cell adhesion was tested on collagen that was first incubated with MMP-1 (versus control on intact collagen). Cell migration on type I collagen was promoted by collagenases. Wound healing of an alveolar epithelial cell monolayer was enhanced in the presence of exogenous collagenases. Our results suggest that collagenases could modulate the repair process by decreasing cell adhesion and cell stiffness, and by increasing cell migration on type I collagen. Collagen degradation could modify cell adhesion sites and collagen degradation peptides could induce alveolar type II pneumocyte migration. New insights regarding alveolar epithelial cell migration are particularly relevant to investigate early events during alveolar epithelial repair following lung injury.

scholarly journals Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

1985 ◽  
Vol 5 (5) ◽  
pp. 1002-1008 ◽  
Author(s):  
E S Allebach ◽  
D Boettiger ◽  
M Pacifici ◽  
S L Adams
Keyword(s):  
Rous Sarcoma Virus ◽  
Type I Collagen ◽  
Skin Fibroblasts ◽  
Transcriptional Level ◽  
Type I ◽  
Type Ii ◽  
Intact Cells ◽  
Rous Sarcoma ◽  
Synthetic Rate ◽  
Sarcoma Virus

We have analyzed the effects of transformation by Rous sarcoma virus on expression of types I and II collagen and fibronectin genes in vertebral chondrocytes and compared them with expression of these genes in skin fibroblasts. Transformed chondrocytes display a dramatically decreased amount of type II collagen RNA, which can account fully for the decreased synthetic rate of this protein. Paradoxically, these cells also display greatly increased amounts of type I collagen RNAs, which are translated efficiently in vitro, but not in the intact cells. We show here that the type I collagen RNAs in transformed chondrocytes are nearly indistinguishable from those found in skin fibroblasts, and that they clearly differ from the type I collagen RNAs found in normal chondrocytes. Transformed chondrocytes also display an increased amount of fibronectin RNAs, which can account fully for the increased synthetic rate of this protein. Thus, the effects of transformation by Rous sarcoma virus on type I collagen and fibronectin RNAs in chondrocytes are the opposite of those observed in fibroblasts, which display decreased amounts of these three RNAs. These data indicate that the effects of transformation on the genes encoding type I collagen and fibronectin must be modulated by host cell-specific factors. They also imply that the types I and II collagen genes may be regulated by different mechanisms, the type I genes being controlled at both transcriptional and posttranscriptional levels, and the type II gene being controlled primarily at the transcriptional level.

Collagen fibrillogenesis in vitro: interaction of types I and V collagen regulates fibril diameter

1990 ◽  
Vol 95 (4) ◽  
pp. 649-657 ◽  
Author(s):  
D.E. Birk ◽  
J.M. Fitch ◽  
J.P. Babiarz ◽  
K.J. Doane ◽  
T.F. Linsenmayer
Keyword(s):  
Type I Collagen ◽  
Small Diameter ◽  
Collagen Molecule ◽  
Type I ◽  
Collagen Types ◽  
Type V Collagen ◽  
Amino Terminal ◽  
Type V ◽  
Fibril Diameter

The small-diameter fibrils of the chick corneal stroma are heterotypic, composed of both collagen types I and V. This tissue has a high concentration of type V collagen relative to other type I-containing tissues with larger-diameter fibrils, suggesting that heterotypic interactions may have a regulatory role in the control of fibril diameter. The interactions of collagen types I and V were studied using an in vitro self-assembly system. Collagens were purified from lathyritic chick embryos in the presence of protease inhibitors. The type V collagen preparations contained higher molecular weight forms of the alpha 1(V) and alpha 2(V) chains constituting 60–70% of the total. Rotary-shadow electron micrographs showed a persistence of a small, pepsin-sensitive terminal region in an amount consistent with that seen by electrophoresis. In vitro, this purified type V collagen formed thin fibrils with no apparent periodicity, while type I collagen fibrils had a broad distribution of large diameters. However, when type I collagen was mixed with increasing amounts of type V collagen a progressive and significant decrease in both the mean fibril diameter and the variance was observed for D periodic fibrils. The amino-terminal domain of the type V collagen molecule was required for this regulatory effect and in its absence little diameter reducing activity was observed. Electron microscopy using collagen type-specific monoclonal antibodies demonstrated that the fibrils formed were heterotypic, containing both collagen types I and V. These data indicate that the interaction of type V with type I collagen is one mechanism modulating fibril diameter and is at least partially responsible for the regulation of collagen fibril formation.

Sign in / Sign up

Export Citation Format

Share Document