Morphological and Molecular Evaluation of the Tissue Repair following Nasal Septum Biopsy in a Sheep Model

Cartilage ◽  
2021 ◽  
pp. 194760352110460
Author(s):  
Maja Pušić ◽  
Matea Brezak ◽  
Andreja Vukasović Barišić ◽  
Mirta Vučković ◽  
Petar Kostešić ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Nasal Septum ◽  
Tissue Repair ◽  
Collagen Type ◽  
Quantitative Polymerase Chain Reaction ◽  
Fibrous Tissue ◽  
Test Group ◽  
Cartilage Tissue ◽  
Type I ◽  
Nasal Cartilage

Objective Nasal septal pathologies requiring surgical intervention are common in the population. Additionally, nasal chondrocytes are becoming an important cell source in cartilage tissue engineering strategies for the repair of articular cartilage lesions. These procedures damage the nasal septal cartilage whose healing potential is limited due to its avascular, aneural, and alymphatic nature. Despite the high incidence of various surgical interventions that affect septum cartilage, limited nasal cartilage repair characterizations have been performed to date. Methods To evaluate the healing of the nasal septum cartilage perforation, a septal biopsy was performed in 14 sheep. Two and 6 months later, the tissue formed on the place of perforation was explanted and compared with the native tissue. Tissue morphology, protein and gene expression of explanted tissue was determined using histological, immunohistochemical and real-time quantitative polymerase chain reaction analysis. Results Tissue formed on the defect site, 2 and 6 months after the biopsy was characterized as mostly connective tissue with the presence of fibroblastic cells. This newly formed tissue contained no glycosaminoglycans and collagen type II but was positively stained for collagen type I. Cartilage-specific genes COL2, AGG, and COMP were significantly decreased in 2- and 6-month samples compared with the native nasal cartilage. Levels of COL1, COL4, and CRABP1 genes specific for perichondrium and connective tissue were higher in both test group samples in comparison with native cartilage. Conclusions Newly formed tissue was not cartilage but rather fibrous tissue suggesting the role of perichondrium and mucosa in tissue repair after nasal septum injury.

Electromagnetic fields enhance chondrogenesis of human adipose-derived stem cells in a chondrogenic microenvironment in vitro

2013 ◽  
Vol 114 (5) ◽  
pp. 647-655 ◽  
Author(s):  
Chung-Hwan Chen ◽  
Yi-Shan Lin ◽  
Yin-Chih Fu ◽  
Chih-Kuang Wang ◽  
Shun-Cheng Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Electromagnetic Field ◽  
Cell Viability ◽  
Chondrogenic Differentiation ◽  
Collagen Type ◽  
Cartilage Tissue ◽  
Pulse Electromagnetic Field ◽  
Type I ◽  
Adipose Derived Stem Cells ◽  
Pemf Treatment

We tested the hypothesis that electromagnetic field (EMF) stimulation enhances chondrogenesis in human adipose-derived stem cells (ADSCs) in a chondrogenic microenvironment. A two-dimensional hyaluronan (HA)-coated well (2D-HA) and a three-dimensional pellet culture system (3D-pellet) were used as chondrogenic microenvironments. The ADSCs were cultured in 2D-HA or 3D-pellet, and then treated with clinical-use pulse electromagnetic field (PEMF) or the innovative single-pulse electromagnetic field (SPEMF) stimulation. The cytotoxicity, cell viability, and chondrogenic and osteogenic differentiations were analyzed after PEMF or SPEMF treatment. The modules of PEMF and SPEMF stimulations used in this study did not cause cytotoxicity or alter cell viability in ADSCs. Both PEMF and SPEMF enhanced the chondrogenic gene expression (SOX-9, collagen type II, and aggrecan) of ADSCs cultured in 2D-HA and 3D-pellet. The expressions of bone matrix genes (osteocalcin and collagen type I) of ADSCs were not changed after SPEMF treatment in 2D-HA and 3D-pellet; however, they were enhanced by PEMF treatment. Both PEMF and SPEMF increased the cartilaginous matrix (sulfated glycosaminoglycan) deposition of ADSCs. However, PEMF treatment also increased mineralization of ADSCs, but SPEMF treatment did not. Both PEMF and SPEMF enhanced chondrogenic differentiation of ADSCs cultured in a chondrogenic microenvironment. SPEMF treatment enhanced ADSC chondrogenesis, but not osteogenesis, when the cells were cultured in a chondrogenic microenvironment. However, PEMF enhanced both osteogenesis and chondrogenesis under the same conditions. Thus the combination of a chondrogenic microenvironment with SPEMF stimulation can promote chondrogenic differentiation of ADSCs and may be applicable to articular cartilage tissue engineering.

scholarly journals Immunochemical and Mechanical Characterization of Cartilage Subtypes in Rabbit

2002 ◽  
Vol 50 (8) ◽  
pp. 1049-1058 ◽  
Author(s):  
Andreas Naumann ◽  
James E. Dennis ◽  
Amad Awadallah ◽  
David A. Carrino ◽  
Joseph M. Mansour ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Joint Analysis ◽  
Type I ◽  
Auricular Cartilage ◽  
Collagen Types ◽  
Type Vi Collagen ◽  
Intense Staining ◽  
Nasal Cartilage

Cartilage is categorized into three general subgroups, hyaline, elastic, and fibrocartilage, based primarily on morphologic criteria and secondarily on collagen (Types I and II) and elastin content. To more precisely define the different cartilage subtypes, rabbit cartilage isolated from joint, nose, auricle, epiglottis, and meniscus was characterized by immunohistochemical (IHC) localization of elastin and of collagen Types I, II, V, VI, and X, by biochemical analysis of total glycosaminoglycan (GAG) content, and by biomechanical indentation assay. Toluidine blue staining and safranin-O staining were used for morphological assessment of the cartilage subtypes. IHC staining of the cartilage samples showed a characteristic pattern of staining for the collagen antibodies that varied in both location and intensity. Auricular cartilage is discriminated from other subtypes by interterritorial elastin staining and no staining for Type VI collagen. Epiglottal cartilage is characterized by positive elastin staining and intense staining for Type VI collagen. The unique pattern for nasal cartilage is intense staining for Type V collagen and collagen X, whereas articular cartilage is negative for elastin (interterritorially) and only weakly positive for collagen Types V and VI. Meniscal cartilage shows the greatest intensity of staining for Type I collagen, weak staining for collagens V and VI, and no staining with antibody to collagen Type X. Matching cartilage samples were categorized by total GAG content, which showed increasing total GAG content from elastic cartilage (auricle, epiglottis) to fibrocartilage (meniscus) to hyaline cartilage (nose, knee joint). Analysis of aggregate modulus showed nasal and auricular cartilage to have the greatest stiffness, epiglottal and meniscal tissue the lowest, and articular cartilage intermediate. This study illustrates the differences and identifies unique characteristics of the different cartilage subtypes in rabbits. The results provide a baseline of data for generating and evaluating engineered repair cartilage tissue synthesized in vitro or for post-implantation analysis.

Is otosclerosis a generalized connective tissue disorder?

1987 ◽  
Vol 101 (4) ◽  
pp. 307-311 ◽  
Author(s):  
H. Oxlund ◽  
U. Pedersen ◽  
C. C. Danielsen ◽  
I. Oxlund ◽  
O. Elbrønd
Keyword(s):  
Connective Tissue ◽  
Collagen Type ◽  
Collagen Type I ◽  
Connective Tissue Disorder ◽  
Biochemical Properties ◽  
Skin Thickness ◽  
Type I ◽  
Molecular Stability ◽  
Cross Links ◽  
Skin Biopsies

AbstractIt has been suggested that otosclerosis might be caused by a generalized disorder in the connective tissue.The biophysical and biochemical properties of skin biopsies from twelve patients with otosclerosis and twelve age- and sex-matched controls were investigated. No differences were found in skin strength and extensibility, skin thickness, collagen content, the relationship between collagen type I and type III, reducible collagen cross-links and molecular stability of collagen type I of samples from patients with otosclerosis as compared with those from the controls. The present study does not support the suggestion that otosclerosis might be a generalized connective tissue disorder.

scholarly journals Morphological features and the functional state of connective tissue of the uterine rudiments in reproductive age patients with Mayer–Rokitansky–Küster–Hauser syndrome

2020 ◽  
Vol 9 (4) ◽  
pp. 24-30
Author(s):  
A.V. Asaturova ◽  
◽  
N.M. Faizullina ◽  
M.V. Bobkova ◽  
A.S. Arakelyan ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Collagen Type ◽  
Tissue Remodeling ◽  
Collagen Type I ◽  
Morphological Features ◽  
Control Group ◽  
Type I ◽  
Variable Degree ◽  
Collagen Iii ◽  
Connective Tissue Remodeling

Introduction. Female patients with Mayer–Rokitansky–Küster–Hauser syndrome (MRKH) have high stigma scores; the condition severely affects the reproductive system. The study aimed at specification of morphological features and assessment of the maturity of connective tissues of the uterine rudiments in MRKH. Patients and methods. The study included 42 patients with vaginal and uterine aplasia having functioning uterine rudiments and 47 patients of the control group without genital malformations. Age of the patients was 20-24 years in 67.2% of the cases, and 31.2% of the patients were aged ≤ 19, inclusive. Immunohistochemi-cal assay was applied to determine expression levels of collagen I, collagen III, ММР2, ММР9, TIMP1, fibronectin and laminin proteins within the functioning uterine rudiments in comparison with levels of the same proteins in normally developed uterine tissues. Results. Decreased expression of collagen type I and elevated levels of MMP2 and MMP9 proteins in uterine tissues were observed for the group of patients with MRKH. Conclusions. 1) Uterine rudiments in patients with MRKH show variable degree of morphological similarity with the normally developed uterus; 2) The functioning uterine rudiments are subject to the same pathological processes as the normally developed uterus (myoma, endometriosis). 3) The functioning uterine rudiments in patients with MRKH show altered patterns of connective tissue remodeling, with decreased expression of collagen type I and increased expression of matrix metalloproteinases MMP2 and MMP9. Keywords: Müllerian aplasia, uterine rudiments, metalloproteinases, connective tissue remodeling, ММР2, ММР9

scholarly journals Connective tissue, glial and neuronal expressions in testis of the African giant rat (Cricetomys gambianus)

2017 ◽  
Vol 34 (03) ◽  
pp. 186-193
Author(s):  
T. Falade ◽  
M. Olude ◽  
O. Mustapha ◽  
E. Mbajiorgu ◽  
A. Ihunwo ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Type I Collagen ◽  
Collagen Type ◽  
Neuronal Cells ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Neuronal Markers ◽  
Type Iii ◽  
African Giant Rat

Abstract Introduction: This study was carried out to investigate the expression of connective tissue (Collagens I and III), glia and neuronal markers in the testis of the African giant rat using histology and immunohistochemistry techniques. Materials and Methods: Eight (8) apparently healthy wild male African giant rats were used for this experiment, divided into 2 groups (juvenile and adult) of 4 animals each. The testes were harvested following intracardial perfusion of the rats and histology was performed using Haematoxylin-Eosin stain and Mallory-Heideinhain rapid one- step staining for connective tissue. Immunohistochemical identification was achieved using the following antibodies: anti-collagen type I, anti-collagen type III, anti-glial fibrillary acidic protein and anti-p75 nerve growth factor for the expression of collagen type I, collagen type III, astrocyte-like cell and neuronal cells respectively. Photomicrography was achieved using Axioskop® microscope and quantitative data were analyzed using student t-test. Results: The cyto-architecture of the testis was typical in the African giant rat. The connective tissue expressed in the juvenile and adult group, signaling of glial-like cells were seen in the perivascular region across the experimental groups. Immuno-localization of neuronal cells were seen in the interstitial spaces across all the groups, but with more expressions in the juvenile. Conclusion: This work has provided a clear description of the expression of connective tissue, neuronal and glial cells in the testis of the African giant rat and their possible relationships across juvenile and adult groups.

scholarly journals Characterization of Collagen Type I and II Blended Hydrogels for Articular Cartilage Tissue Engineering

2016 ◽  
Vol 17 (10) ◽  
pp. 3145-3152 ◽  
Author(s):  
Nelda Vázquez-Portalatı́n ◽  
Claire E. Kilmer ◽  
Alyssa Panitch ◽  
Julie C. Liu
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Cartilage Tissue Engineering ◽  
Collagen Type I ◽  
Cartilage Tissue ◽  
Type I

scholarly journals Adipose-Derived Mesenchymal Stem Cell Chondrospheroids Cultured in Hypoxia and a 3D Porous Chitosan/Chitin Nanocrystal Scaffold as a Platform for Cartilage Tissue Engineering

2020 ◽  
Vol 21 (3) ◽  
pp. 1004 ◽  
Author(s):  
Veronica Zubillaga ◽  
Ana Alonso-Varona ◽  
Susana C. M. Fernandes ◽  
Asier M. Salaberria ◽  
Teodoro Palomares
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Collagen Type ◽  
Porous Scaffold ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Type I ◽  
Low Oxygen ◽  
Porous Chitosan

Articular cartilage degeneration is one of the most common causes of pain and disability in middle-aged and older people. Tissue engineering (TE) has shown great therapeutic promise for this condition. The design of cartilage regeneration constructs must take into account the specific characteristics of the cartilaginous matrix, as well as the avascular nature of cartilage and its cells’ peculiar arrangement in isogenic groups. Keeping these factors in mind, we have designed a 3D porous scaffold based on genipin-crosslinked chitosan/chitin nanocrystals for spheroid chondral differentiation of human adipose tissue-derived mesenchymal stem cells (hASCs) induced in hypoxic conditions. First, we demonstrated that, under low oxygen conditions, the chondrospheroids obtained express cartilage-specific markers including collagen type II (COL2A1) and aggrecan, lacking expression of osteogenic differentiation marker collagen type I (COL1A2). These results were associated with an increased expression of hypoxia-inducible factor 1α, which positively directs COL2A1 and aggrecan expression. Finally, we determined the most suitable chondrogenic differentiation pattern when hASC spheroids were seeded in the 3D porous scaffold under hypoxia and obtained a chondral extracellular matrix with a high sulphated glycosaminoglycan content, which is characteristic of articular cartilage. These findings highlight the potential use of such templates in cartilage tissue engineering.

scholarly journals Assessment of carvacrol paste as endodontic medication associated with laser photobiomodulation in tissue repair: an experimental study in rats

2020 ◽  
Vol 36 (5) ◽  
Author(s):  
Ismário Silva Meneses ◽  
Ricardo Luiz Cavalcanti de Albuquerque Júnior ◽  
Felipe de Souza Matos ◽  
Aline Aragão Pereira Macedo ◽  
Adriano Antunes de Souza Araújo ◽  
...  
Keyword(s):  
Tissue Repair ◽  
Wistar Rats ◽  
Type I Collagen ◽  
Subcutaneous Tissue ◽  
Fibrous Tissue ◽  
Tissue Reaction ◽  
Type Iii Collagen ◽  
Type I ◽  
Subcutaneous Implantation ◽  
Inflammatory Infiltration

This study aimed to analyze the tissue reaction caused by carvacrol paste associated or not with laser photobiomodulation (LPBM) at λ660 nm in the subcutaneous tissue of rats. Sixty Wistar rats were divided into four groups and they received the following interventions: subcutaneous implantation of empty polyethylene tubes (CTR), implantation of tubes containing carvacrol paste (CVC), implantation of empty tubes and LPBM (LLLT), and implantation of tubes containing carvacrol paste and LPBM (CVCLT). The animals were euthanized at three, eight, and 15 days after surgery. The inflammatory reaction and fibroplasia were analyzed histomorphometrically. Significant differences among the groups were determined by ANOVA and Tukey's test (p<0.05). In the 3-day period, the CVCLT group had low inflammatory infiltration (p<0.01). In the 8- and 15-day periods, the LLLT and CVCLT groups presented a low amount of lymphocytic inflammatory infiltrate (p<0.01 and p<0.05). Regarding the formation of fibrous tissue, the CVC group had the highest formation of type III collagen in the 8-day period (p<0.001). In the 15-day period, the CVCLT group had a lower formation of type I collagen than the CTR and LLLT groups (p<0.05). The use of the carvacrol paste associated with photobiomodulation optimizes the inflammatory period and tissue repair.

EFFECT OF PURIFIED ALGINATE MICROCAPSULES ON THE REGENERATION OF CHONDROCYTES

2012 ◽  
Vol 24 (03) ◽  
pp. 185-195 ◽  
Author(s):  
Ji Hye Hwang ◽  
On You Kim ◽  
A Ram Kim ◽  
Ji Yeon Bae ◽  
Su Mi Jeong ◽  
...  
Keyword(s):  
Cell Viability ◽  
Collagen Type ◽  
Cartilage Regeneration ◽  
Collagen Type I ◽  
Tissue Growth ◽  
Cartilage Tissue ◽  
Type I ◽  
Hematoxylin And Eosin ◽  
Safranin O

Adult articular cartilage tissue has poor capability of self-repair. Therefore, a variety of tissue engineering approaches are motivated by the clinical need for articular repair. Alginate has been used as a biomaterial for cartilage regeneration. The alginate is a natural polymer that is extracted from seaweeds and purification. However, the main drawback is the immune rejection in vivo. To overcome this problem, we have developed the biocompability of alginate using modified Korbutt method. After alginate was purified, purified alginate microcapsules were used in cartilage regeneration. Chondrocytes were seeded in purified and nonpurified alginate microcapsules, and then cell viability, proliferation and phenotype were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to confirm mRNA expression on collagen type I and collagen type II for chondrocytes phenotype. Hematoxylin and eosin (H&E) and Safranin-O histological staining showed tissue growth at the interface during the first 10 days. In this study, chondrocytes in purified alginate microcapsules had higher cell viability, proliferation and more phenotype expression than those in nonpurified alginate microcapsules. The results suggest that the purified alginate microcapsule is useful for cartilage regeneration.

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