scholarly journals Tissue-Engineering the Fibrous Pancreatic Tumour Stroma Capsule in 3D Tumouroids to Demonstrate Paclitaxel Response

2021 ◽  
Vol 22 (8) ◽  
pp. 4289
Author(s):  
Judith Pape ◽  
Katerina Stamati ◽  
Rawiya Al Hosni ◽  
Ijeoma F. Uchegbu ◽  
Andreas G. Schatzlein ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Drug Testing ◽  
Collagen Type I ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type I ◽  
Tumour Mass ◽  
Stromal Compartment ◽  
Pancreatic Tumour ◽  
Initial Drug

Pancreatic cancer is a unique cancer in that up to 90% of its tumour mass is composed of a hypovascular and fibrotic stroma. This makes it extremely difficult for chemotherapies to be delivered into the core of the cancer mass. We tissue-engineered a biomimetic 3D pancreatic cancer (“tumouroid”) model comprised of a central artificial cancer mass (ACM), containing MIA Paca-2 cells, surrounded by a fibrotic stromal compartment. This stromal compartment had a higher concentration of collagen type I, fibronectin, laminin, and hyaluronic acid (HA) than the ACM. The incorporation of HA was validated with alcian blue staining. Response to paclitaxel was determined in 2D MIA Paca-2 cell cultures, the ACMs alone, and in simple and complex tumouroids, in order to demonstrate drug sensitivity within pancreatic tumouroids of increasing complexity. The results showed that MIA Paca-2 cells grew into the complex stroma and invaded as cell clusters with a maximum distance of 363.7 µm by day 21. In terms of drug response, the IC50 for paclitaxel for MIA Paca-2 cells increased from 0.819 nM in 2D to 3.02 nM in ACMs and to 5.87 nM and 3.803 nM in simple and complex tumouroids respectively, indicating that drug penetration may be significantly reduced in the latter. The results demonstrate the need for biomimetic models during initial drug testing and evaluation.

Thyroid Follicular Hyperplasia Associated With Massive Extracellular Mucin Deposition

2017 ◽  
Vol 25 (6) ◽  
pp. 533-535 ◽  
Author(s):  
Francesco Nesa ◽  
Luca Poggi ◽  
Stefano Ferrero ◽  
Alessandro Del Gobbo
Keyword(s):  
Alcian Blue ◽  
Periodic Acid ◽  
The Other ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Thyroid Tumors ◽  
Stromal Compartment ◽  
Follicular Hyperplasia ◽  
Nodular Hyperplasia ◽  
Thyroid Parenchyma

Extensive extracellular mucin deposition is a rare pathological thyroid condition with 6 cases described in literature so far. We report another case of a 67-year-old woman, discussing histopathological features, and review the literature. Our findings showed a diffuse mucin deposition in the stromal compartment of thyroid parenchyma. Histochemical stainings showed positivity for Alcian blue staining, but not for periodic acid–Shiff staining. Our case is peculiar because this mucin deposition was associated with benign nodular hyperplasia, in contrast with the other 6 reports, which described the same stromal alterations associated with benign or malignant thyroid tumors.

Wound-healing effects of human dermal components with gelatin dressing

2017 ◽  
Vol 32 (6) ◽  
pp. 716-724 ◽  
Author(s):  
Hyun-Jun Jang ◽  
Yu-mi Kim ◽  
Bo-Young Yoo ◽  
Young-Kwon Seo
Keyword(s):  
Wound Healing ◽  
Normal Skin ◽  
Treated Group ◽  
Collagen Type I ◽  
Skin Wound ◽  
Blue Staining ◽  
Elastic Fibers ◽  
Type I ◽  
Initial Area

There have been numerous investigations regarding various types of dressings and artificial dermis of solid form, yet limited research and development on paste types, such as hydrogels with dermal powder, have ensued. In this study, we compared the in vivo wound healing effects of gelatin paste containing dermal powder to a collagen type I/chondroitin 6-sulfate (coll/chondroitin) sponge and gelatin alone, after 48 days post grafting, in a skin wound rat model. In the dermis powder/gelatin paste-treated group, wound area contraction was minimized 50%, while in the gelatin and coll/chondroitin sponge groups, the initial area contracted 83–85% and 79–85%, respectively. Histological analysis revealed the wounds treated with dermal powder/gelatin were associated with many fibroblasts, which infiltrated the wound bed, as well as thick collagen bundles that were arranged in dendritic arrays, resembling normal skin. Furthermore, in contrast to the gelatin- and coll/chondroitin sponge-treated groups, the powder/gelatin paste-treated wounds exhibited an abundance of elastic fibers (Victoria blue staining) and extensive formation of blood vessels around the dermis (CD31 staining). Therefore, the dermis powder/gelatin paste not only renders convenience to users but also has prominent wound-healing effects on full-thickness wounds.

scholarly journals Collagen type I-induced Smad-interacting protein 1 expression downregulates E-cadherin in pancreatic cancer

Oncogene ◽  
2006 ◽  
Vol 26 (16) ◽  
pp. 2381-2385 ◽  
Author(s):  
Y Imamichi ◽  
A König ◽  
T Gress ◽  
A Menke
Keyword(s):  
Pancreatic Cancer ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Interacting Protein ◽  
E Cadherin

scholarly journals Use of adipose stem cells and polylactide discs for tissue engineering of the temporomandibular joint disc

2009 ◽  
Vol 7 (42) ◽  
pp. 177-188 ◽  
Author(s):  
Katja Mäenpää ◽  
Ville Ellä ◽  
Jari Mauno ◽  
Minna Kellomäki ◽  
Riitta Suuronen ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Temporomandibular Joint ◽  
Adipose Stem Cells ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type I ◽  
Mrna Levels ◽  
Chondrogenic Medium ◽  
Tmj Disc

There is currently no suitable replacement for damaged temporomandibular joint (TMJ) discs after discectomy. In the present study, we fabricated bilayer biodegradable polylactide (PLA) discs comprising a non-woven mat of poly(L/D)lactide (P(L/D)LA) 96/4 and a P(L/DL)LA 70/30 membrane plate. The PLA disc was examined in combination with adipose stem cells (ASCs) for tissue engineering of the fibrocartilaginous TMJ disc in vitro . ASCs were cultured in parallel in control and chondrogenic medium for a maximum of six weeks. Relative expression of the genes, aggrecan, type I collagen and type II collagen present in the TMJ disc extracellular matrix increased in the ASC-seeded PLA discs in the chondrogenic medium. The hypertrophic marker, type X collagen, was moderately induced. Alcian blue staining showed accumulation of sulphated glycosaminoglycans. ASC differentiation in the PLA discs was close to that observed in pellet cultures. Comparison of the mRNA levels revealed that the degree of ASC differentiation was lower than that in TMJ disc-derived cells and tissue. The pellet format supported the phenotype of the TMJ disc-derived cells under chondrogenic conditions and also enhanced their hyalinization potential, which is considered part of the TMJ disc degeneration process. Accordingly, the combination of ASCs and PLA discs has potential for the development of a tissue-engineered TMJ disc replacement.

scholarly journals A 3D-Printed PLCL Scaffold Coated with Collagen Type I and Its Biocompatibility

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Yong He ◽  
Wei Liu ◽  
Lianxiong Guan ◽  
Jielin Chen ◽  
Li Duan ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Mechanical Test ◽  
Great Influence ◽  
Collagen Type I ◽  
Blue Staining ◽  
Type I ◽  
3D Printed ◽  
Better Than

Scaffolds play an important role in tissue engineering and their structure and biocompatibility have great influence on cell behaviors. In this study, poly(l-lactide-co-ε-caprolactone) (PLCL) scaffolds were printed by a 3D printing technology, low-temperature deposition manufacturing (LDM), and then PLCL scaffolds were treated by alkali and coated with collagen type I (COLI). The scaffolds were characterized by scanning electron microscopy (SEM), porosity test, mechanical test, and infrared spectroscopy. The prepared PLCL and PLCL-COLI scaffolds had three-dimensional (3D) porous structure and they not only have macropores but also have micropores in the deposited lines. Although the mechanical property of PLCL-COLI was slightly lower than that of PLCL scaffold, the hydrophilicity of PLCL-COLI was significantly enhanced. Rabbit articular chondrocytes were extracted and were identified as chondrocytes by toluidine blue staining. To study the biocompatibility, the chondrocytes were seeded on scaffolds for 1, 3, 5, 7, and 10 days. MTT assay showed that the proliferation of chondrocytes on PLCL-COLI scaffold was better than that on PLCL scaffold. And the morphology of cells on PLCL-COLI after 1-day culture was much better than that on PLCL. This 3D-printed PLCL scaffold coated with COLI shows a great potential application in tissue engineering.

scholarly journals OP0199 EX VIVO BIOMARKER PROFILING IDENTIFIES ONCOSTATIN-M AS SPINE OSTEOARTHRITIS-SPECIFIC OSTEOIMMUNOLOGICAL TARGET

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 120.2-120
Author(s):  
O. Roessinger ◽  
M. Blanchard ◽  
M. Briki ◽  
D. Jurić ◽  
C. Netzer ◽  
...  
Keyword(s):  
Facet Joint ◽  
Ex Vivo ◽  
Oncostatin M ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type I ◽  
Tgf Beta ◽  
Disease Heterogeneity ◽  
Knee Oa ◽  
Cartilage Proteoglycans

Background:Disease heterogeneity, both clinically and molecularly, has been a major hurdle in the development of efficacious disease-modifying osteoarthritis drugs (DMOADs). Biomechanical, inflammatory, osteoporotic and metabolic OA have been proposed as clinically relevant subtypes for stratification of knee OA patients, yet this remains to be included in clinical trial design. Disease heterogeneity does not only occur within, but also between joint types. However, robust data on joint-specific pathomechanisms of OA are still lacking.Objectives:In this study, we performed ex vivo biomarker profiling of human osteochondral tissue of knee and spine OA to identify joint-specific pathomechanisms and DMOAD treatment responses.Methods:Facet joint and tibial plateaus were obtained from patients undergoing lumbar spinal fusion (n=11, mean age 72.8) and total joint arthroplasty (n=8, mean age 73.0) respectively. Osteochondral specimens were cut in equal-sized samples (100-300 mg wet weight) and randomly assigned to treatment groups: control (DMSO), inflammation (1 μg/mL LPS) or inflammation + DMOAD (TGF-betatype I receptor inhibitor,10 μM SB-505124). Explant culture was conducted for one week and biomarkers of bone metabolism (Pro-Col-Ia, SOST, OPG, SPP1), inflammation (MCP-1, IL-6, MMP3, OSM, TIMP1, VEGFA) and cartilage metabolism (ACAN, COMP) were determined by ELISA. Normalized biomarker secretion was analysed using clusteranalyses and ANOVA. Cartilage proteoglycans were assessed by whole mount Alcian blue staining. Expression of Oncostatin-M (OSM) and its receptors OSMR and LIFR in joint tissues was assessed by RT-PCR and immunohistochemistry.Results:Clusteranalyses revealed that LPS stimulation increased IL-6 and MCP-1 secretion by both facet joint (FJ) and knee joint (KJ) tissues. Interestingly, Oncostatin-M (OSM) and its downstream mediators MMP3 and TIMP1 were increased in the majority of FJ, but not KJ specimens. Statistical analyses corroborated increased OSM, MMP3 and TIMP1 levels in a spine-specific fashion (Figure).Whole mount Alcian blue staining revealed heterogeneous effects of LPS treatment on cartilage proteoglycans, which was negatively correlated with OSM (r=-0.54) and TIMP1 levels (r=-0.45) – yet poorly associated with ACAN (r=0.19). Inhibition of TGF-beta type I receptor signalling in osteochondral tissues led to a drastic reduction of Pro-Collagen-Ia and IL-6 secretion in both spine and knee OA specimens. Interestingly, DMOAD treatment significantly reduced OSM, TIMP1 and MMP3 levels in FJ specimens only. Vice versa, KJ tissues revealed a specific upregulation of monocyte chemoattractant protein-1 (MCP-1) and osteopontin (SPP1) upon inhibition of TGF-beta signalling. OSM was exclusively expressed in subchondral bone marrow macrophages. Isolated chondrocytes and osteoblasts expressed both LIFR and OSMR, yet intact cartilage only showed OSMR expression, while OSMR and LIFR was expressed in marrow tissueConclusion:Oncostatin-M expression and signalling was uncovered as specific pathomechanism of spine OA. DMOAD treatment effects suggested interplay of OSM and TGF-beta signalling pathways in facet joint osteoarthritis. Known to be predominantly expressed by macrophages and immune cells, OSM may be an important osteoimmunological mediator of tissue damage and remodelling in spine, but not knee OA. This study also highlights the value of ex vivo human tissue models for OA phenotyping and preclinical evaluation of DMOADs.Disclosure of Interests:None declared

scholarly journals Histochemistry of Equine Damaged Tendons, Ligaments and Articular Cartilage

2018 ◽  
Vol 46 (1) ◽  
pp. 8
Author(s):  
Gabriela De Bastiani ◽  
Flávio Desessards De La Corte ◽  
Karin Erica Brass ◽  
Camila Cantarelli ◽  
Stefano Dau ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Connective Tissue ◽  
Alcian Blue ◽  
Collagen Type ◽  
Repair Process ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type Iii Collagen ◽  
Type I ◽  
Loose Connective Tissue

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.

scholarly journals Uniaxial Static Strain Promotes Osteoblast Proliferation and Bone Matrix Formation in Distraction Osteogenesis In Vitro

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Zhengqiang Li ◽  
Junfa Zheng ◽  
Di Wan ◽  
Xiaoqin Yang
Keyword(s):  
Distraction Osteogenesis ◽  
Mechanical Stimulation ◽  
Collagen Type ◽  
Bone Matrix ◽  
Collagen Type I ◽  
Blue Staining ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Static Strain ◽  
Stimulation Of

Objective. We aimed at investigating the effects of uniaxial static strain on osteoblasts in distraction osteogenesis (DO). Methods. To simulate the mechanical stimulation of osteoblasts during DO, 10% uniaxial static strain was applied to osteoblasts using a homemade multiunit cell stretching and compressing device. Before and after applying strain stimulation, the morphological changes of osteoblasts were observed by inverted phase-contrast microscopy, Coomassie blue staining, and immunofluorescence. Alkaline phosphatase (ALP) activity, mRNA levels (proliferating cell nuclear antigen [PCNA], ALP, Runx2, osteocalcin [OCN], collagen type I, hypoxia-inducible factor- [HIF-] 1α, and vascular endothelial growth factor [VEGF]), and protein levels (Runx2, OCN, collagen type I, HIF-1α, and VEGF) were evaluated by using ALP kit, real-time quantitative reverse transcription-polymerase chain reaction, western blot, and enzyme-linked immunosorbent assay. Results. After the mechanical stimulation, the cytoskeleton microfilaments were rearranged, and the cell growth direction of the osteoblasts became ordered, with their direction being at an angle of about 45° from the direction of strain. The proliferation of osteoblasts and the expression levels of mRNA and protein of ALP, Runx2, OCN, collagen type I, HIF-1α, and VEGF were significantly higher than in the nonstretch control groups. Conclusion. Our homemade device can exert uniaxial static strain and promote the proliferation of osteoblasts and bone matrix formation. It can be used to simulate the mechanical stimulation of osteoblasts during DO.

scholarly journals Evaluation of a Cell-Free Collagen Type I-Based Scaffold for Articular Cartilage Regeneration in an Orthotopic Rat Model

Materials ◽  
2020 ◽  
Vol 13 (10) ◽  
pp. 2369 ◽  
Author(s):  
Marta Anna Szychlinska ◽  
Giovanna Calabrese ◽  
Silvia Ravalli ◽  
Anna Dolcimascolo ◽  
Paola Castrogiovanni ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Collagen Type ◽  
Cartilage Regeneration ◽  
Collagen Type I ◽  
Collagen I ◽  
Host Cells ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Type I ◽  
A Cell

The management of chondral defects represents a big challenge because of the limited self-healing capacity of cartilage. Many approaches in this field obtained partial satisfactory results. Cartilage tissue engineering, combining innovative scaffolds and stem cells from different sources, emerges as a promising strategy for cartilage regeneration. The aim of this study was to evaluate the capability of a cell-free collagen I-based scaffold to promote cartilaginous repair after orthotopic implantation in vivo. Articular cartilage lesions (ACL) were created at the femoropatellar groove in rat knees and cell free collagen I-based scaffolds (S) were then implanted into right knee defect for the ACL-S group. No scaffold was implanted for the ACL group. At 4-, 8- and 16-weeks post-transplantation, degrees of cartilage repair were evaluated by morphological, histochemical and gene expression analyses. Histological analysis shows the formation of fibrous tissue, at 4-weeks replaced by a tissue resembling the calcified one at 16-weeks in the ACL group. In the ACL-S group, progressive replacement of the scaffold with the newly formed cartilage-like tissue is shown, as confirmed by Alcian Blue staining. Immunohistochemical and quantitative real-time PCR (qRT-PCR) analyses display the expression of typical cartilage markers, such as collagen type I and II (ColI and ColII), Aggrecan and Sox9. The results of this study display that the collagen I-based scaffold is highly biocompatible and able to recruit host cells from the surrounding joint tissues to promote cartilaginous repair of articular defects, suggesting its use as a potential approach for cartilage tissue regeneration.

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