scholarly journals Anatomical and histological analyses reveal that tail repair is coupled with regrowth in wild-caught, juvenile American alligators (Alligator mississippiensis)

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Cindy Xu ◽  
Joanna Palade ◽  
Rebecca E. Fisher ◽  
Cameron I. Smith ◽  
Andrew R. Clark ◽  
...  

AbstractReptiles are the only amniotes that maintain the capacity to regenerate appendages. This study presents the first anatomical and histological evidence of tail repair with regrowth in an archosaur, the American alligator. The regrown alligator tails constituted approximately 6–18% of the total body length and were morphologically distinct from original tail segments. Gross dissection, radiographs, and magnetic resonance imaging revealed that caudal vertebrae were replaced by a ventrally-positioned, unsegmented endoskeleton. This contrasts with lepidosaurs, where the regenerated tail is radially organized around a central endoskeleton. Furthermore, the regrown alligator tail lacked skeletal muscle and instead consisted of fibrous connective tissue composed of type I and type III collagen fibers. The overproduction of connective tissue shares features with mammalian wound healing or fibrosis. The lack of skeletal muscle contrasts with lizards, but shares similarities with regenerated tails in the tuatara and regenerated limbs in Xenopus adult frogs, which have a cartilaginous endoskeleton surrounded by connective tissue, but lack skeletal muscle. Overall, this study of wild-caught, juvenile American alligator tails identifies a distinct pattern of wound repair in mammals while exhibiting features in common with regeneration in lepidosaurs and amphibia.

2017 ◽  
Vol 122 (3) ◽  
pp. 533-540 ◽  
Author(s):  
Abigail L. Mackey ◽  
Michael Kjaer

Human skeletal muscle has the potential to regenerate completely after injury induced under controlled experimental conditions. The events inside the myofibers as they undergo necrosis, followed closely by satellite cell-mediated myogenesis, have been mapped in detail. Much less is known about the adaptation throughout this process of both the connective tissue structures surrounding the myofibers and the fibroblasts, the cells responsible for synthesizing this connective tissue. However, the few studies investigating muscle connective tissue remodeling demonstrate a strong response that appears to be sustained for a long time after the major myofiber responses have subsided. While the use of electrical stimulation to induce eccentric contractions vs. voluntary eccentric contractions appears to lead to a greater extent of myofiber necrosis and regenerative response, this difference is not apparent when the muscle connective tissue responses are compared, although further work is required to confirm this. Pharmacological agents (growth hormone and angiotensin II type I receptor blockers) are considered in the context of accelerating the muscle connective tissue adaptation to loading. Cautioning against this, however, is the association between muscle matrix protein remodeling and protection against reinjury, which suggests that a (so far undefined) period of vulnerability to reinjury may exist during the remodeling phases. The role of individual muscle matrix components and their spatial interaction during adaptation to eccentric contractions is an unexplored field in human skeletal muscle and may provide insight into the optimal timing of rest vs. return to activity after muscle injury.


2017 ◽  
Vol 313 (1) ◽  
pp. R44-R50 ◽  
Author(s):  
Shivam H. Patel ◽  
Andrew C. D’Lugos ◽  
Erica R. Eldon ◽  
Donald Curtis ◽  
Jared M. Dickinson ◽  
...  

Acetaminophen (APAP) given during chronic exercise reduces skeletal muscle collagen and cross-linking in rats. We propose that the effect of APAP on muscle extracellular matrix (ECM) may, in part, be mediated by dysregulation of the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs). The purpose of this study was to evaluate the impact of APAP consumption during acute resistance exercise (RE) on several regulators of the ECM in human skeletal muscle. In a double-blinded, placebo-controlled, randomized crossover design, recreationally active men ( n = 8, 25 ± 2 yr) performed two trials of knee extension. Placebo (PLA) or APAP (1,000 mg/6 h) was given for 24 h before and immediately following RE. Vastus lateralis biopsies were taken at baseline and 1 and 3 h post-RE. Quantitative RT-PCR was used to determine differences in mRNA expression. MMP-2, type I collagen, and type III collagen mRNA expression was not altered by exercise or APAP ( P > 0.05). When compared with PLA, TIMP-1 expression was lower at 1 h post-RE during APAP conditions but greater than PLA at 3 h post-RE ( P < 0.05). MMP-9 expression and protein levels were elevated at 3 h post-RE independent of treatment ( P < 0.05). Lysyl oxidase expression was greater at 3 h post-RE during APAP consumption ( P < 0.05) compared with PLA. MMP-2 and TIMP-1 protein was not altered by RE or APAP ( P > 0.05). Phosphorylation of ERK1/2 and p38-MAPK increased ( P < 0.05) with RE but was not influenced by APAP. Our findings do not support our hypothesis and suggest that short-term APAP consumption before RE has a small impact on the measured ECM molecules in human skeletal muscle following acute RE.


1984 ◽  
Vol 219 (3) ◽  
pp. 1017-1026 ◽  
Author(s):  
N Light ◽  
A E Champion

In the past it has been proven difficult to separate and characterize collagen from muscle because of its relative paucity in this tissue. The present report presents a comprehensive methodology, combining methods previously described by McCollester [(1962) Biochim. Biophys. Acta 57, 427-437] and Laurent, Cockerill, McAnulty & Hastings [(1981) Anal. Biochem. 113, 301-312], in which the three major tracts of muscle connective tissue, the epimysium, perimysium and endomysium, may be prepared and separated from the bulk of muscle protein. Connective tissue thus prepared may be washed with salt and treated with pepsin to liberate soluble native collagen, or can be washed with sodium dodecyl sulphate to produce a very clean insoluble collagenous product. This latter type of preparation may be used for quantification of the ratio of the major genetic forms of collagen or for measurement of reducible cross-link content to give reproducible results. It was shown that both the epimysium and perimysium contain type I collagen as the major component and type III collagen as a minor component; perimysium also contained traces of type V collagen. The endomysium, the sheaths of individual muscle fibres, was shown to contain both type I and type III collagen as major components. Type V collagen was also present in small amounts, and type IV collagen, the collagenous component of basement membranes, was purified from endomysial preparations. This is the first biochemical demonstration of the presence of type IV collagen in muscle endomysium. The preparation was shown to be very similar to other type IV collagens from other basement membranes on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and was indistinguishable from EHS sarcoma collagen and placenta type IV collagen in the electron microscope after rotary shadowing.


2020 ◽  
Vol 10 (4) ◽  
pp. 381-389
Author(s):  
Sergey V. Minaev ◽  
Sergey I. Timofeev ◽  
Alina N. Grigorova ◽  
Oksana V. Vladimirova ◽  
Elena I. Pashneva ◽  
...  

Aim. To assess adhesions of the abdominal cavity in children with varying degrees of connective tissue dysplasia (CTD) severity. Methods. A total of 91 children with average age of 10.6 1.4 years were observed from 2005 to 2019. Composed of 53 boys (58.2%) and 38 girls (41.8%). Patients were divided into two groups: group 1 (69 children without CTD) and group 2 (22 children with CTD). Clinical data and screening cards were used to diagnose DST. All patients underwent an assessment for the clinical course, a macroscopic visual assessment adhesive process severity in the abdominal cavity according to three different scales (Nair, F. Coccolini, N.I. Ayushinova). Immunohistochemistry was performed against collagen I, III, IV type, Laminin 1, angiogenesis factors, transforming growth factor beta (TGF-), and Fibroblast Growth Factor Receptor 1 (FGFR1). A semi-quantitative counting method was used to record the relative number of immunopositive structures. Statistical analysis was carried out by methods of variation statistics using the Chi-square test and MannWhitney U-tests, as well as the Spearman rank correlation method for the reliability of differences between groups. Results. In the adhesive process visualization, the N.I. Ayushinov scale was revealed to be the most informative, showing the average score of 7 0.8 points in group 2 and 14 1.2 in group 1 (rs 0.35; p 0.05). In group 2, the adhesive material showed no (56%) or chaotic fragments (44%) of type I collagen, and the ratio of type I to type III collagen is 2.7: 5.1. In group 1, the ratio of type I to type III collagen is 5.9:1.8. The ratio of collagen IV to type I is 6.5: 2.9. Both groups have a moderate (++) amount of TGF-. TGF- is positive with macrophages. FGFR1 was found in the control group (++++). A positive response was seen in fibroblasts and macrophages (U = 79.00; p = 0.006). Statistically significant analyzes of vascular endothelial growth factor in compared groups (rs = 0.632, p 0.001) had a positive correlation.. Conclusion. Thus, the study showed features of clinical course and morphological changes during the development of adhesions in the abdominal cavity in children with varying degrees of severity of CTD. Data obtained dictate the need for an individual approach in predicting adhesive disease, as well as targeted preventive care.


2006 ◽  
Vol 386 (7-8) ◽  
pp. 1961-1966 ◽  
Author(s):  
Cyril Petibois ◽  
Gilles Gouspillou ◽  
Katia Wehbe ◽  
Jean-Paul Delage ◽  
Gérard Déléris

Author(s):  
Viktor A. Lazarenko ◽  
Sergey V. Ivanov ◽  
Ilya S. Ivanov ◽  
Evgeny G. Ob’edkov ◽  
Nikolay N. Grigor'ev ◽  
...  

The aim is to examine the effectiveness of preoperative skin biopsy method to determine indications for preventive endoprosthesis implantation of the anterior abdominal wall based on structural features of connective tissue. Materials and methods. The study included 17 patients who had been planned to undergo a scheduled surgery such as video laparoscopic cholecystectomy. The histological material was collected with DERMO PUNCH biopsy needles in an outpatient clinic. Using polarized light microscopy we inspected histological slices coloured with Sirius red. After that we analyzed histological sections including the ratio of collagen fibers, the density of connective tissue and the number of interfibrous spaces. Results. All the studied features of connective tissue dysplasia had been registered among 18% of patients; significant differences (p ≤0.05) had been identified among other 24% of patients. In patients with identified signs of connective tissue dysplasia, an increase in type III collagen fibers and a decrease in type I fibers were revealed, which directly resulted in a decrease in their ratio. In addition, this category of patients noted a decrease in the density of connective tissue and a significant number of interfibrous spaces. Conclusion. Skin biopsy is a safe and informative method to reveal patients with predisposition to the formation of hernias of the anterior abdominal wall based on structural features of connective tissue and to determine indications for preventive endoprosthetic implantation. This method will reduce the frequency of postoperative ventral hernias, so the effectiveness of the treatment will definitely increase.


1987 ◽  
Vol 42 (11-12) ◽  
pp. 1303-1306 ◽  
Author(s):  
Waltraud Folkhard ◽  
Daniela Christmann ◽  
Werner Geercken ◽  
Ernst Knörzer ◽  
Michel H. J. Koch ◽  
...  

X-ray diffraction analysis of connective tissue samples, which contain type I and type III collagen shows that twisted collagen fibrils are a general principle of assembly. The occurrence of twisted fibrils in native wet Chordae tendineae, skin and Aorta is combined with a shorter axial periodicity of about 65 nm. This shorter D period is shown to be directly related to the tilt of the molecules, which have to be curved to build-up twisted fibrils.


2018 ◽  
Vol 46 (1) ◽  
pp. 8
Author(s):  
Gabriela De Bastiani ◽  
Flávio Desessards De La Corte ◽  
Karin Erica Brass ◽  
Camila Cantarelli ◽  
Stefano Dau ◽  
...  

Background: The injury repair process in tendons and ligaments includes different phases such as inflammation, neovascularization, fibroblast proliferation and fibrosis. Collagen type and tissue characteristics of tendon and ligament repair are described such as type collagen differentiation and properties of the scars tissue. The degeneration of articular cartilage when, characterized by loss of the articular layers associated of the decreased of proteoglycans. The aim of this study is to describe by histochemistry techniques the characteristics of tissue scar, collagen type in the repair process of tendons and ligaments, as well as articular cartilage degeneration.Materials, Methods & Results: Tissue samples of equine tendons, ligaments and articular cartilage of the metacarpophalangeal joint region were evaluated by ultrasonography, macroscopically and prepared for routine histopathology (H&E staining). The inclusion criterion of the samples in this study was based on the presence of lesions characterized in H&E stain as fibroplasia, neovascularization, collagenolysis, chondroid metaplasia in tendons and ligaments and fibrillation and cartilaginous eburnation lesions in the articular cartilage samples. The Masson’s trichrome, Picrosirius red and Alcian blue staining techniques were also performed in addition to H&E. Pathologic findings in the tendons and ligaments included fibroplasia, collagenolysis, chondroid metaplasia and lymphohistioplasmacytic inflammation. Tendons and ligaments scars were composed of type III collagen but there was also some type I collagen. Fiber alignment of tendons and ligaments in the reorganization tissue was not flawless and the fiber appearance was characterized by a lack of the fiber crimp and parallelism. The fibroplasia was characterized by endotendinous tickening areas associated with the presence of loose connective tissue. In the areas of loose connective tissue substitution, collagen type fibers are intercalated to a lesser extent by type-III collagen fibers. In the Alcian blue stained samples of articular cartilage observed the surface layer and the matrix zone of calcified cartilage were weakly stained in blue.Discussion: Three special stains were utilized in this study along with the H&E evaluation elucidating the behavior tendons, ligaments and articular cartilage injury. The important observation in this study was fibroplasia in tendons and ligaments seems to be composed by abundant of loose connective tissue, chondrocytes and intermingled collagen type I and III fibers associated with lack of crimps alignment of the fibers. The fragile structure suggested by the Masson’s trichrome stain results (presence of the loose connective tissue) in this study perhaps make the tendons and ligaments receptive to other lesions. The characteristic blue discoloration of collagen fibers was only observed in the loose connective tissue may be because the dye penetration becomes easier when compared to the dense connective tissue (stained in red). The Masson’s trichrome made possible the differentiated the dense connective tissue of the loose connective tissue. The combined histochemistry staining technics allowed an improved characterization of fiber alignment, collagen type, inflammatory cell infiltration and neovascularization, which happens during the repair process of tendons and ligaments. The fibrillation and eburnation of the articular cartilage were associated with the decrease Alcian Blue staining characterized by degeneration process of articular cartilage.


Author(s):  
E.M. Kuhn ◽  
K.D. Marenus ◽  
M. Beer

Fibers composed of different types of collagen cannot be differentiated by conventional electron microscopic stains. We are developing staining procedures aimed at identifying collagen fibers of different types.Pt(Gly-L-Met)Cl binds specifically to sulfur-containing amino acids. Different collagens have methionine (met) residues at somewhat different positions. A good correspondence has been reported between known met positions and Pt(GLM) bands in rat Type I SLS (collagen aggregates in which molecules lie adjacent to each other in exact register). We have confirmed this relationship in Type III collagen SLS (Fig. 1).


Diabetes ◽  
1985 ◽  
Vol 34 (8) ◽  
pp. 812-815 ◽  
Author(s):  
L. Borghi ◽  
R. Lugari ◽  
A. Montanari ◽  
P. Dall'Argine ◽  
G. F. Elia ◽  
...  

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