scholarly journals Comparative Anatomy of Axial Skeleton of Red-eared Turtle (Trachemys scripta elegans, Wied 1838) and Softshell Turtle (Amyda cartilaginea, Boddaert 1770)

2019 ◽  
Vol 2 ◽  
pp. 97-100
Author(s):  
Hanif Mustafa ◽  
Muhammad Ja’far Luthfi ◽  
Fadhilatul Ilmi ◽  
Ida Khoirunnisa ◽  
Takrima Takrima
Keyword(s):  
Comparative Anatomy ◽  
Cervical Vertebrae ◽  
Axial Skeleton ◽  
Trachemys Scripta ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Lateral Side ◽  
Trachemys Scripta Elegans ◽  
Alizarin Red ◽  
Softshell Turtles

Red-eared turtle and softshell turtles belong Cryptodira Suborder which has a different characteristic in neck length and head movement. The aim of this study was to determine of the axial skeleton anatomical structure including vertebrae, carapace and plastron of the red-eared turtle (Trachemys scripta elegans Wied, 1838) and softshell turtles (Amyda cartilaginea Boddaert, 1770) females. This research was carried out for five months starting from September 2013 to January 2014. The methods used in this study were th e X-Ray method, boiled bone and Alizarin Red S-Alcian blue staining. The result of research was analyzed descriptively comparatively by direct observation using a digital camera. Based on the results of the study the Red-eared turtle tortoise has a number of 7 cervical vertebrae, 9th vertebrae, sacral vertebrae 1 segment and vertebrae caudalis 27 segments. The anterior and posterior zygapophysis of the cervix elongate thus affecting the limited lateral movement. The thoracic center of the vertebrae adjusts the shape of the carapace. The sacralis vertebrae have 1 centrum segment extending on the lateral side attached to the carapace called the lateral pars, the caudal centrum is short and there is a shortened anterior zygapophysis structure. Whereas softshell turtles   have slender and long centrums. The anterior and posterior zygapophysis are smaller and allow the softshell turtles to perform more lateral movements. Centrum vertebrae of the thorachalis have a flat shape adjusting the shape of the carapace. Sacralis vertebrae have 2 centrum and 2 lateral pars extending and meeting each other to form a hole sacralia pelvina, centrum vertebrae caudalis extends and there is a neural spinal structure. Carapace of the red-eared turtle consists of fused pieces. Whereas the carapace in the softshell turtles consists of pieces covered by cartilage. The constituent component of carapace and plastron of the red-eared turtle consists of true bones completely, while the constituent components of the carapace and plastron of softshell turtles consist of true bones and cartilage on the sides and connective between the carapace and plastron.

scholarly journals Anatomical Study of Red-eared TurteTail (Trachemys scripta elegans)

2019 ◽  
Vol 2 ◽  
pp. 137-140
Author(s):  
Putri Nofita ◽  
Muhammad Ja’far Luthfi ◽  
Reza Sukma Dewi
Keyword(s):  
Anatomical Study ◽  
Slow Motion ◽  
Axial Skeleton ◽  
Trachemys Scripta ◽  
Histological Structure ◽  
Fat Tissue ◽  
Trachemys Scripta Elegans ◽  
Alizarin Red ◽  
First Time ◽  
Hematoxylin Eosin

Turtles are reptiles armored backs hard, slow motion, appearing for the first time about 200 million years ago and relative ly unchanged for 150 million years. This study aims to determine the anatomical structure of the axial skeleton and determine the histologic structure of red-eared turtles (Trachemys scripta elegans) females. The method used, among others, Alizarin Red S staining Alcian Blue, methods of paraffin with Hematoxylin-eosin staining (HE) and Mallory acid fuchsin and used 3 adult turtles 3 years old. The results showed that vertebral caudalis red-eared turtles (Trachemys scripta elegans) females composed of true bone, vertebrae-type amphicoelous not have plains autotomy and histology red-eared turtles (Trachemys scripta elegans) females resemble histological structure other tail reptiles with some differences that only have 4 files do not have the muscle and fat tissue.

scholarly journals Alizarin Red S-Alcian Blue Staining for Regenerated tail of Common House Gecko (Hemidactylus frenatus)

2018 ◽  
Vol 7 (2) ◽  
pp. 57-59
Author(s):  
Rakhmiyati Rakhmiyati ◽  
Muhammad Ja’far Luthfi
Keyword(s):  
Alcian Blue ◽  
Histochemical Staining ◽  
Alizarin Red S ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Tail Regeneration ◽  
Alizarin Red ◽  
Regenerate Tail ◽  
Hemidactylus Frenatus ◽  
Common House

Common House Gecko (Hemidactylus frenatus) is one of reptiles that have ability to autotomy their tails. Tail autotomy is a mechanism to protect it self from predators. After the tail broke, there will be wound healing on the tail which is then followed by a tail regeneration event. Original tail and regenerate tail is very different morphologically and anatomically. The original tail is composed of bones while the tail of the regenerate is composed of cartilage. Histochemical staining using Alizarin Red-S Alcian Blue was done to differentiate bone and cartilage. This method will stained bones red while the cartilage will stained blue.

scholarly journals LncRNA DANCR and miR-320a suppressed osteogenic differentiation in osteoporosis by directly inhibiting the Wnt/β-catenin signaling pathway

2020 ◽  
Vol 52 (8) ◽  
pp. 1310-1325
Author(s):  
Cheng-Gong Wang ◽  
Yi-He Hu ◽  
Shi-Long Su ◽  
Da Zhong
Keyword(s):  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Morphological Changes ◽  
Osteoporosis Treatment ◽  
Inhibitory Effect ◽  
Luciferase Assay ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Alizarin Red

Abstract Our study aimed to determine how lncRNA DANCR, miR-320a, and CTNNB1 interact with each other and regulate osteogenic differentiation in osteoporosis. qRT-PCR and western blotting were performed to determine the expression of DANCR, miR-320a, CTNNB1, and the osteoporosis- or Wnt/β-catenin pathway-related markers T-cell factor 1 (TCF-1), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). Interactions between CTNNB1, DANCR, and miR-320a were predicted by bioinformatics approaches and validated using a luciferase assay. Osteoblastic phenotypes were evaluated by ALP staining, ALP activity assay and Alizarin Red staining. The bilateral ovariectomy method was used to establish an in vivo osteoporosis model. Bone morphological changes were examined using hematoxylin and eosin (H&E) and Alcian Blue staining. The expression levels of DANCR and miR-320a in BMSCs derived from osteoporosis patients were upregulated, whereas CTNNB1 expression was downregulated compared with that in healthy controls. Importantly, we demonstrated that miR-320a and DANCR acted independently from each other and both inhibited CTNNB1 expression, whereas the inhibitory effect was additive when miR-320a and DANCR were cooverexpressed. Moreover, we found that DANCR overexpression largely abrogated the effect of the miR-320a inhibitor on CTNNB1 expression and the Wnt/β-catenin signaling pathway in BMSCs during osteogenic differentiation. We further confirmed the results above in BMSCs derived from an osteoporosis animal model. Taken together, our findings revealed that DANCR and miR-320a regulated the Wnt/β-catenin signaling pathway during osteogenic differentiation in osteoporosis through CTNNB1 inhibition. Our results highlight the potential value of DANCR and miR-320a as promising therapeutic targets for osteoporosis treatment.

scholarly journals Modified Alizarin Red S-Alcian Blue Staining for Reptilian Skeleton

2016 ◽  
Vol 5 (1) ◽  
pp. 19
Author(s):  
Muhammad Ja’far Luthfi
Keyword(s):  
Alcian Blue ◽  
Anatomical Study ◽  
Alizarin Red S ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Double Staining ◽  
Alizarin Red ◽  
Important Method ◽  
Clearing Method ◽  
And Cartilage

<p>Skeletal staining is an important method in anatomical study. The aim of the research was to develop staining and clearing method of Reptilian skeleton using Alizarin Red S-Alcian Blue. The specimen were eviscerated, fixed, stained, cleared, and keep in glycerine solution. This modified double-staining has successfully stain bone and cartilage of Reptilian.</p>

scholarly journals The effect of aging on the biological and immunological characteristics of periodontal ligament stem cells

2020 ◽  
Author(s):  
Xiaoyu Li ◽  
Bowen Zhang ◽  
Hong Wang ◽  
Xiaolu Zhao ◽  
Zijie Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Periodontal Ligament ◽  
Chondrogenic Differentiation ◽  
Annexin V ◽  
Cell Counting ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Periodontal Ligament Stem Cells ◽  
Alizarin Red ◽  
Differentiation Potential

Abstract Background: Periodontal ligament stem cells (PDLSCs) have many applications in the field of cytotherapy, tissue engineering, and regenerative medicine. However, the effect of age on the biological and immunological characteristics of PDLSCs remains unclear. Methods: In this study, we compared PDLSCs isolated from young and adult individuals. PDLSCs proliferation was analyzed by Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) staining, and apoptosis level was detected by Annexin V-PE/7-ADD staining. PDLSCs osteogenic/adipogenic/chondrogenic differentiation potentials were assessed by alkaline phosphatase (ALP), Alizarin Red, Oil Red O, Alcian Blue staining and related quantitative analysis. PDLSCs immunosuppressive capacity was determined by EdU and Annexin V-PE/7-ADD staining. To explore its underlying mechanism, microarray, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), and western blot analyses were performed to detect differentially expressed genes and proteins in PDLSCs. Results: Our results demonstrated that with aging, the proliferation and osteogenic/adipogenic/chondrogenic differentiation potential of PDLSCs decreased, whereas apoptosis of PDLSCs increased. Moreover, the immunosuppressive ability of PDLSCs decreased with aging. Compared with PDLSCs from young subjects, analysis of mRNA expression revealed an upregulation of CCND3 and RC3H2, and a downregulation of Runx2, ALP, COL1A1, PPARγ2, CXCL12, FKBP1A, FKBP1B, NCSTN, P2RX7, PPP3CB, RIPK2, SLC11A1, and TP53 in those from adult individuals. Furthermore, protein expression levels of Runx2, ALP, COL1A1, and PPARγ2 in the adult group was decreased, whereas that of CCND3 increased. Conclusions: Taken together, aging influences biological and immunological characteristics of PDLSCs, and thus it is more appropriate to utilized PDLSCs from young individuals for tissue regeneration, post-aging treatment, and allotransplantation.

308 ISOLATION, DIFFERENTIATION, AND IMMUNOPHENOTYPIC CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM EQUINE ADIPOSE TISSUE AND BONE MARROW

2011 ◽  
Vol 23 (1) ◽  
pp. 250
Author(s):  
E. Iacono ◽  
B. Merlo ◽  
A. Spadari ◽  
G. Mari ◽  
F. Ricci ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Cellular Therapy ◽  
Population Doubling ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Alizarin Red ◽  
Minimum Criteria

Minimum criteria for the characterisation of human mesenchymal stem cells (MSC) are a) adhesion to the plastic when maintained under culture conditions; b) expression of CD105, CD73, and CD90, and no expression for CD45, CD34, and CD14; and c) differentiation into osteoblasts, adipocytes, and chondroblasts in vitro. One major difficulty in characterising equine MSC is the absence of specific monoclonal antibodies and evidence that certain markers from other species do not cross-react with the equine species. The aim of this work was to isolate, cultivate, differentiate, and conduct cellular characterisation of MSC derived from equine adipose tissue (AT) and bone marrow (BM). Adipose tissue collection was performed at the base of the horses’ tails, and BM was aspirated from the iliac crest. Mononuclear cell fraction was isolated and cultured as previously described by (Colleoni et al. 2009 Vet. Res. Commun. 33, 811–821). Chondrogenic, osteogenic, and adipogenic differentiation were performed in monolayer culture, and evidence for differentiation was made by morphological and cytological evaluations. For molecular characterisation, cells were treated with trypsin, washed with PBS, and fixed with Reagent 1 (Intraprep Kit, Beckman Coulter, Miami, FL, USA), following the manufacturer’s instructions. Samples, after washings, were incubated for 20 min at room temperature with CD105, CD90, CD44, CD45, CD34, CD14, and CD73 mAbs, directly conjugated to fluorescein isothiocyanate, PE, or APC (Beckman Coulter). Appropriate conjugate isotype controls were applied (Beckman Coulter). After staining, cells were washed twice with PBS, and fluorescence intensity was evaluated with a FC500 two-laser equipped cytometer (Beckman Coulter). Results were further analysed with the CXP dedicated program. Samples volumes were 68 ± 23.6 mL for BM and 5.6 ± 1.1 g for AT; in both AT and BM, the isolation rate was 100% (AT: 4/4; BM: 5/5). Undifferentiated cells were passaged up to 8 times for AT and 5 times for BM; population-doubling times (DT) were calculated, and data were analysed by ANOVA (Statistica for Windows, Stat Soft Inc., Tulsa, OK, USA). No significant differences (P > 0.05) were found between DT of all passages. The DT was greater (P < 0.05) for BM (3.2 ± 1.5) than for AT (1.3 ± 0.7). By passage 8, AT MSC underwent 37.3 ± 4.6 cell-doublings (CD); by passage 5, BM MSC underwent 26.2 ± 5.03 CD. Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian blue staining illustrated chondrogenesis, and positive Oil Red O staining suggested adipogenesis. The AT and BM MSC were positive for CD90, CD44, and CD105; all cell lines were negative for haematopoietic markers such as CD34, CD14, and CD45. Although marker CD73 expresses reaction in other studies involving MSC in different species, it did not cross-react with equine AT and BM MSC. Results obtained revealed the immunophenotypic characterisation of the surface of isolated and cultivated MSC, classifying these cells as a promising type of progenitor cells that can be applied in equine cellular therapy.

Mineralization in the syrinx and caudal tracheal rings in the ostrich

2017 ◽  
Author(s):  
Sukru Hakan Atalgin ◽  
Sevinc Ates ◽  
Ibrahim Kurtul ◽  
Hakan Terzi
Keyword(s):  
Alcian Blue ◽  
Bone Cells ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Double Staining ◽  
Alizarin Red ◽  
Struthio Camelus ◽  
Calcium Accumulation ◽  
Microscopic Features

This study documented macroscopic and microscopic features and mineralization of the caudally located tracheal rings and syrinx in two ostrich (struthio camelus) having three years of age. The syrinx and trachea of the birds were stained in toto with alcian blue and alizarin red for cartilage and mineralization. Observations on the syrinx and trachea, measurements and photography were performed under stereo-microscopy. They were stained grossly using alizarin red and alcian blue to visualize mineralization and histologically by hemotoxylin and eosin (HandE) to detect ossification areas, if any. Results revealed incomplete tracheal rings located caudally in-between the intact ones. Alizarin red and alcian blue staining displayed mineralized regions grossly. Histological slides by HandE staining showed no ossification. Overall results proposed that alizarin red and alcian blue double staining is a good toll to determine mineralization which is calcium accumulation in the tissue before the formation of bone cells.

scholarly journals Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

2015 ◽  
Vol 65 (1) ◽  
pp. 66-78 ◽  
Author(s):  
FRANCUSKI V Jelena ◽  
DEBELJAK MARTAČIĆ Jasmina ◽  
RADOVANOVIĆ Anita ◽  
ANDRIĆ Nenad ◽  
SOURICE-PETIT Sophie ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Three Dimensional ◽  
Population Doubling ◽  
Blue Staining ◽  
Population Doubling Time ◽  
Alcian Blue Staining ◽  
German Shepherd ◽  
Alizarin Red ◽  
Differentiation Potential

Abstract The aim of this study was to determine whether synovial fluid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37°C, 5% CO2, humidified atmosphere) three weeks before their phenotype was characterized by flow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets’ slides have shown a significant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.

210 COMPARATIVE CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM MINIATURE PIG SYNOVIUM, SYNOVIAL FLUID AND BONE MARROW

2012 ◽  
Vol 24 (1) ◽  
pp. 217 ◽  
Author(s):  
W. J. Lee ◽  
G. H. Maeng ◽  
R. H. Jeon ◽  
G. J. Rho ◽  
S. L. Lee
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Miniature Pig ◽  
Rt Pcr ◽  
Alizarin Red ◽  
Differentiated Cells ◽  
Oil Red O Staining ◽  
Oil Red O

Mesenchymal stem cells (MSC) are a valuable cell source for cartilage tissue engineering because MSC derived from various tissues readily differentiate into skeletal cell and chondrogenic lineages. In this study, we compared the cellular characteristics of synovium (SN) and synovial fluid (SF)-derived MSC with bone marrow (BM)-derived MSC of miniature pig. Biopsies of SN and BM were collected and SF was flushed with saline solution from 6-month-old miniature pig (T-type, PWG Micro-pig, PWG Genetics Korea, South Korea). Cells were isolated from collected tissues and cultured in advanced-DMEM supplemented with 10% fetal bovine serum at 38.5°C in a humidified atmosphere of 5% CO2 in air. The cells were then evaluated for their expression of MSC-specific markers, including CD29, CD44 and CD90 using flow cytometry. The expression of early transcriptional factors, such as Oct3/4, Nanog and Sox2 was evaluated by immunocytochemical staining and reverse transcription-PCR (RT-PCR). Multilineage differentiation of MSC were induced under conditions conductive for osteogenic, adipogenic and chondrogenic lineages and then evaluated by von Kossa and Alizarin Red S staining, Oil red O staining and Alcian Blue staining, respectively. Differentiated cells were further analysed for the expression of lineage specific markers by RT-PCR. Statistical analysis was performed using one-way ANOVA by SPSS. The SN-, SF- and BM-MSC were observed to be positive for MSC specific markers, such as CD29 (99 ± 0.2, 96 ± 0.5 and 98 ± 0.2, respectively), CD44 (97 ± 0.3, 96 ± 0.6 and 98 ± 0.5, respectively) and CD90 (95 ± 0.5, 92 ± 0.2 and 96 ± 0.3, respectively); however, haematopoietic marker CD45 (2.0 ± 2.1, 3.0 ± 1.3 and 2.0 ± 3.0, respectively) was barely detected. In all MSC, early transcription factors (Oct3/4, Nanog and Sox2) were expressed by immnocytochemical staining and the transcripts were detected by RT-PCR. Following exposure to the specific differentiation medium, all these MSC were capable of differentiating into mesenchymal lineages, such as osteogenic, adiopogenic and chondrogenic as assessed by von Kossa and Alizarin Red S staining, Oil red O staining and and Alcian Blue staining, respectively. In addition, differentiated cells from all MSC expressed the marker genes specific to osteocytes (osteonectin, Runx2), adipocytes (aP2, PRAR-γ2) and chondrocytes (aggrecan, collagen type 2) by RT-PCR. The results of this study suggested that cells isolated from miniature pig articular tissues, such as SN and SF have similar characteristics of MSCs and their differentiation ability was comparable to BM-MSC. Hence, it is possible to establish MSCs from SN and SF as alternate sources during the biopsy of synovial tissues for arthritis diagnosis. Further studies are being carried out to evaluate their in vivo differentiation potential into chondrocytes. This study was supported by Rural Development Administration (grant No. 20110701-305-533-001-02-00) and National Research Foundation of Korea (grant No. 2011-0010252) of the Republic of Korea.

scholarly journals STRUKTUR TULANG DAN OTOT SIRIP KAUDAL KOMPLEKS Andamia heteroptera Bleeker (IKAN AMFIBI)

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Gatot Nugroho Susanto
Keyword(s):  
Alcian Blue ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Alizarin Red ◽  
Caudal Fin ◽  
Primitive Type

Andamia heteroptera, known as rockskipper fish, is one of amphibious fish. Most of amphibious fish use caudal fin to facilitate locomotion in land and water. The modification is fusion of os hypural and reduction-fusion os epural and uroneurals. The aim of this study was to  acces skeleton and musculature structure of A. heteroptera caudal fin. Alizarin red-Alcian blue staining revealed that caudal fin of A. heteroptera has wider spina hemalis and spina neuralis support the locomotion. There is modification of caudal fin structure for support the locomotion. Caudal fin musculature of A. heteroptera is identified as primitive type, since its lack of musculus hypochordal longitudinalis. 

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