scholarly journals Validation of the optimal scaffold pore size of nasal implants using the 3-dimensional culture technique

2020 ◽  
Vol 47 (4) ◽  
pp. 310-316
Author(s):  
Jeoung Hyun Nam ◽  
So Yun Lee ◽  
Galina Khan ◽  
Eun Soo Park
Keyword(s):  
Extracellular Matrix ◽  
Pore Size ◽  
Culture Technique ◽  
Patient Specific ◽  
Remarkable Difference ◽  
Cell Seeding ◽  
3 Dimensional ◽  
Viable Cells ◽  
Porous Disc ◽  
Number Of Cells

Background To produce patient-specific nasal implants, it is necessary to harvest and grow autologous cartilage. It is crucial to the proliferation and growth of these cells for scaffolds similar to the extracellular matrix to be prepared. The pore size of the scaffold is critical to cell growth and interaction. Thus, the goal of this study was to determine the optimal pore size for the growth of chondrocytes and fibroblasts.Methods Porous disc-shaped scaffolds with 100-, 200-, 300-, and 400-µm pores were produced using polycaprolactone (PCL). Chondrocytes and fibroblasts were cultured after seeding the scaffolds with these cells, and morphologic evaluation was performed on days 2, 14, 28, and 56 after cell seeding. On each of those days, the number of viable cells was evaluated quantitatively using an MTT assay.Results The number of cells had moderately increased by day 28. This increase was noteworthy for the 300- and 400-µm pore sizes for fibroblasts; otherwise, no remarkable difference was observed at any size except the 100-µm pore size for chondrocytes. By day 56, the number of cells was observed to increase with pore size, and the number of chondrocytes had markedly increased at the 400-µm pore size. The findings of the morphologic evaluation were consistent with those of the quantitative evaluation.Conclusions Experiments using disc-type PCL scaffolds showed (via both morphologic and quantitative analysis) that chondrocytes and fibroblasts proliferated most extensively at the 400-µm pore size in 56 days of culture.

scholarly journals Patient-Specific 3-Dimensional Model of Smooth Muscle Cell and Extracellular Matrix Dysfunction for the Study of Aortic Aneurysms

2021 ◽  
pp. 152660282110092
Author(s):  
Natalija Bogunovic ◽  
Jorn P. Meekel ◽  
Jisca Majolée ◽  
Marije Hekhuis ◽  
Jakob Pyszkowski ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Time Course ◽  
Spatial Organization ◽  
Specific Model ◽  
Pharmacological Therapy ◽  
Aortic Aneurysms ◽  
Patient Specific ◽  
Preclinical Model ◽  
3 Dimensional

Introduction: Abdominal aortic aneurysms (AAAs) are associated with overall high mortality in case of rupture. Since the pathophysiology is unclear, no adequate pharmacological therapy exists. Smooth muscle cells (SMCs) dysfunction and extracellular matrix (ECM) degradation have been proposed as underlying causes. We investigated SMC spatial organization and SMC-ECM interactions in our novel 3-dimensional (3D) vascular model. We validated our model for future use by comparing it to existing 2-dimensional (2D) cell culture. Our model can be used for translational studies of SMC and their role in AAA pathophysiology. Materials and Methods: SMC isolated from the medial layer of were the aortic wall of controls and AAA patients seeded on electrospun poly-lactide- co-glycolide scaffolds and cultured for 5 weeks, after which endothelial cells (EC) are added. Cell morphology, orientation, mechanical properties and ECM production were quantified for validation and comparison between controls and patients. Results: We show that cultured SMC proliferate into multiple layers after 5 weeks in culture and produce ECM proteins, mimicking their behavior in the medial aortic layer. EC attach to multilayered SMC, mimicking layer interactions. The novel SMC model exhibits viscoelastic properties comparable to biological vessels; cytoskeletal organization increases during the 5 weeks in culture; increased cytoskeletal alignment and decreased ECM production indicate different organization of AAA patients’ cells compared with control. Conclusion: We present a valuable preclinical model of AAA constructed with patient specific cells with applications in both translational research and therapeutic developments. We observed SMC spatial reorganization in a time course of 5 weeks in our robust, patient-specific model of SMC–EC organization and ECM production.

Use of human aortic extracellular matrix as a scaffold for construction of a patient-specific tissue engineered vascular patch

2017 ◽  
Vol 12 (6) ◽  
pp. 065006 ◽  
Author(s):  
Li-Ping Gao ◽  
Ming-Jun Du ◽  
Jing-Jing Lv ◽  
Sebastian Schmull ◽  
Ri-Tai Huang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Patient Specific ◽  
Specific Tissue

scholarly journals Mesenchymal Stem Cell Seeding of Porcine Small Intestinal Submucosal Extracellular Matrix for Cardiovascular Applications

PLoS ONE ◽  
2016 ◽  
Vol 11 (4) ◽  
pp. e0153412 ◽  
Author(s):  
Chia Wei Chang ◽  
Tye Petrie ◽  
Alycia Clark ◽  
Xin Lin ◽  
Claus S. Sondergaard ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cell Seeding ◽  
Small Intestinal

Generation of 2.5D lung bud organoid from human induced pluripotent stem cell

2021 ◽  
pp. 1-14
Author(s):  
Xun Xu ◽  
Yan Nie ◽  
Weiwei Wang ◽  
Imran Ullah ◽  
Wing Tai Tung ◽  
...  
Keyword(s):  
Single Cell ◽  
Disease Modeling ◽  
3D Culture ◽  
Induced Pluripotent Stem Cell ◽  
Patient Specific ◽  
Homogeneous Distribution ◽  
Cell Seeding ◽  
Differentiation Potential ◽  
Induced Pluripotent ◽  
Defined Culture

Human induced pluripotent stem cells (hiPSCs) are a promising cell source to generate the patient-specific lung organoid given their superior differentiation potential. However, the current 3D cell culture approach is tedious and time-consuming with a low success rate and high batch-to-batch variability. Here, we explored the establishment of lung bud organoids by systematically adjusting the initial confluence levels and homogeneity of cell distribution. The efficiency of single cell seeding and clump seeding was compared. Instead of the traditional 3D culture, we established a 2.5D organoid culture to enable the direct monitoring of the internal structure via microscopy. It was found that the cell confluence and distribution prior to induction were two key parameters, which strongly affected hiPSC differentiation trajectories. Lung bud organoids with positive expression of NKX 2.1, in a single-cell seeding group with homogeneously distributed hiPSCs at 70%confluence (SC_70%_hom) or a clump seeding group with heterogeneously distributed cells at 90%confluence (CL_90%_het), can be observed as early as 9 days post induction. These results suggest that a successful lung bud organoid formation with single-cell seeding of hiPSCs requires a moderate confluence and homogeneous distribution of cells, while high confluence would be a prominent factor to promote the lung organoid formation when seeding hiPSCs as clumps. 2.5D organoids generated with defined culture conditions could become a simple, efficient, and valuable tool facilitating drug screening, disease modeling and personalized medicine.

scholarly journals Injectable, redox-polymerized carboxymethylcellulose hydrogels promote nucleus pulposus-like extracellular matrix elaboration by human MSCs in a cell density-dependent manner

2018 ◽  
Vol 33 (4) ◽  
pp. 576-589 ◽  
Author(s):  
Devika M Varma ◽  
Michelle S DiNicolas ◽  
Steven B Nicoll
Keyword(s):  
Extracellular Matrix ◽  
Minimally Invasive ◽  
Nucleus Pulposus ◽  
Seeding Density ◽  
Human Mscs ◽  
Dependent Manner ◽  
Cell Seeding ◽  
Swelling Properties ◽  
Matrix Deposition ◽  
Cell Seeding Density

Low back pain is a major cause for disability and is closely linked to intervertebral disc degeneration. Mechanical and biological dysfunction of the nucleus pulposus in the disc has been found to initiate intradiscal degenerative processes. Replacing or enriching the diseased nucleus pulposus with an injectable, stem cell-laden biomaterial that mimics its material properties can provide a minimally invasive strategy for biological and structural repair of the tissue. In this study, injectable, in situ-gelling carboxymethylcellulose hydrogels were developed for nucleus pulposus tissue engineering using encapsulated human marrow-derived mesenchymal stromal cells (hMSCs). With the goal of obtaining robust extracellular matrix deposition and faster construct maturation, two cell-seeding densities, 20 × 106 cells/ml and 40 × 106 cells/ml, were examined. The constructs were fabricated using a redox initiation system to yield covalently crosslinked, cell-seeded hydrogels via radical polymerization. Chondrogenic culture of the hydrogels over 35 days exhibited high cell viability along with deposition of proteoglycan and collagen-rich extracellular matrix, and mechanical and swelling properties similar to native human nucleus pulposus. Further, the matrix production and distribution in the carboxymethylcellulose hydrogels was found to be strongly influenced by hMSC-seeding density, with the lower cell-seeding density yielding a more favorable nucleus pulposus-specific matrix phenotype, while the rate of construct maturation was less dependent on the cell-seeding density. These findings are the first to demonstrate the utility of redox-polymerized carboxymethylcellulose hydrogels as hMSC carriers for potential minimally invasive treatment strategies for nucleus pulposus replacement.

Variation in femoral anteversion in patients requiring total hip replacement

2019 ◽  
Vol 30 (3) ◽  
pp. 281-287
Author(s):  
Jim W Pierrepont ◽  
Ed Marel ◽  
Jonathan V Baré ◽  
Leonard R Walter ◽  
Catherine Z Stambouzou ◽  
...  
Keyword(s):  
Total Hip Replacement ◽  
Hip Replacement ◽  
Femoral Stem ◽  
Femoral Anteversion ◽  
Patient Specific ◽  
3 Dimensional ◽  
Total Hip ◽  
Age Related ◽  
Gender And Age ◽  
Implant Alignment

Background: Optimal implant alignment is important for total hip replacement (THR) longevity. Femoral stem anteversion is influenced by the native femoral anteversion. Knowing a patient’s femoral morphology is therefore important when planning optimal THR alignment. We investigated variation in femoral anteversion across a patient population requiring THR. Methods: Preoperatively, native femoral neck anteversion was measured from 3-dimensional CT reconstructions in 1215 patients. Results: The median femoral anteversion was 14.4° (−27.1–54.5°, IQR 7.4–20.9°). There were significant gender differences (males 12.7°, females 16.0°; p < 0.0001). Femoral anteversion in males decreased significantly with increasing age. 14% of patients had extreme anteversion (<0° or >30°). Conclusions: This is the largest series investigating native femoral anteversion in a THR population. Patient variation was large and was similar to published findings of a non-THR population. Gender and age-related differences were observed. Native femoral anteversion is patient-specific and should be considered when planning THR.

scholarly journals Endothelial Cell Seeding onto the Extracellular Matrix of Fibroblasts for the Development of a Small Diameter Polyurethane Vessel

ASAIO Journal ◽  
1993 ◽  
Vol 39 (3) ◽  
pp. M740-M745 ◽  
Author(s):  
Yoon-Shin Lee ◽  
Dong Kook Park ◽  
Yong Bae Kim ◽  
Jeong Wook Seo ◽  
Kyu Back Lee ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cell ◽  
Small Diameter ◽  
Cell Seeding ◽  
Endothelial Cell Seeding

scholarly journals Stimulation of corneal differentiation by interaction between cell surface and extracellular matrix. I. Morphometric analysis of transfilter "induction".

1975 ◽  
Vol 66 (2) ◽  
pp. 275-291 ◽  
Author(s):  
L Meier ◽  
E D Hay
Keyword(s):  
Extracellular Matrix ◽  
Electron Microscope ◽  
Cell Surface ◽  
Pore Size ◽  
Collagen Synthesis ◽  
Lens Capsule ◽  
Physical Contact ◽  
Corneal Epithelial ◽  
Cell Processes

The present study was undertaken to determine whether or not physical contact with the substratum is essential for the stimulatory effect of extracellular matrix (ECM) on corneal epithelial collagen synthesis. Previous studies showed that collagenous substrata stimulate isolated epithelia to produce three times as much collagen as they produce on noncollagenous substrate; killed collagenous substrata (e.g., lens capsule) are just as effective as living substrata (e.g., living lens) in promoting the production of new corneal stroma in vitro. In the experiments to be reported here, corneal epithelia were placed on one side of Nucleopore filters of different pore sizes and killed lens capsule on the other, with the expectation that contact of the reacting cells with the lens ECM should be limited by the number and size of the cell processes that can tranverse the pores. Transfilter cultures were grown for 24 h in [3H]proline-containing median and incorporation of isotope into hot trichloroacetic acid-soluble protein was used to measure corneal epithelial collagen production. Epithelial collagen synthesis increases directly as the size of the pores in the interposed filter increases and decreases as the thickness of the filter layer increases. Cell processes within Nucleopore filters were identified with the transmission electron microscope with difficulty; with the scanning electron microscope, however, the processes could easily be seen emerging from the undersurface of even 0.1-mum pore size filters. Morphometric techniques were used to show that cell surface area thus exposed to the underlying ECM is linearly correlated with enhancement of collagen synthesis. Epithelial cell processes did not pass through ultrathin (25-mum thick) 0.45-mum pore size Millipore filters nor did "induction" occur across them. The results are discussed in relation to current theories of embryonic tissue interaction.

scholarly journals On-Demand Intraoperative 3-Dimensional Printing of Custom Cranioplastic Prostheses

2018 ◽  
Vol 15 (3) ◽  
pp. 341-349 ◽  
Author(s):  
Alexander I Evins ◽  
John Dutton ◽  
Sayem S Imam ◽  
Amal O Dadi ◽  
Tao Xu ◽  
...  
Keyword(s):  
3D Printing ◽  
Perioperative Complications ◽  
Ease Of Use ◽  
Patient Specific ◽  
Production Time ◽  
3 Dimensional ◽  
3D Print ◽  
Fused Deposition ◽  
On Demand ◽  
Dimensional Printing

Abstract BACKGROUND Currently, implantation of patient-specific cranial prostheses requires reoperation after a period for design and formulation by a third-party manufacturer. Recently, 3-dimensional (3D) printing via fused deposition modeling has demonstrated increased ease of use, rapid production time, and significantly reduced costs, enabling expanded potential for surgical application. Three-dimensional printing may allow neurosurgeons to remove bone, perform a rapid intraoperative scan of the opening, and 3D print custom cranioplastic prostheses during the remainder of the procedure. OBJECTIVE To evaluate the feasibility of using a commercially available 3D printer to develop and produce on-demand intraoperative patient-specific cranioplastic prostheses in real time and assess the associated costs, fabrication time, and technical difficulty. METHODS Five different craniectomies were each fashioned on 3 cadaveric specimens (6 sides) to sample regions with varying topography, size, thickness, curvature, and complexity. Computed tomography-based cranioplastic implants were designed, formulated, and implanted. Accuracy of development and fabrication, as well as implantation ability and fit, integration with exiting fixation devices, and incorporation of integrated seamless fixation plates were qualitatively evaluated. RESULTS All cranioprostheses were successfully designed and printed. Average time for design, from importation of scan data to initiation of printing, was 14.6 min and average print time for all cranioprostheses was 108.6 min. CONCLUSION On-demand 3D printing of cranial prostheses is a simple, feasible, inexpensive, and rapid solution that may help improve cosmetic outcomes; significantly reduce production time and cost—expanding availability; eliminate the need for reoperation in select cases, reducing morbidity; and has the potential to decrease perioperative complications including infection and resorption.

scholarly journals What Have In Vitro Co-Culture Models Taught Us about the Contribution of Epithelial-Mesenchymal Interactions to Airway Inflammation and Remodeling in Asthma?

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1694
Author(s):  
Emmanuel Twumasi Osei ◽  
Steven Booth ◽  
Tillie-Louise Hackett
Keyword(s):  
Extracellular Matrix ◽  
Cell Proliferation ◽  
Airway Inflammation ◽  
Airway Epithelial ◽  
Disease Pathogenesis ◽  
3 Dimensional ◽  
Lung Epithelial ◽  
Culture Models

As the lung develops, epithelial-mesenchymal crosstalk is essential for the developmental processes that drive cell proliferation, differentiation, and extracellular matrix (ECM) production within the lung epithelial-mesenchymal trophic unit (EMTU). In asthma, a number of the lung EMTU developmental signals have been associated with airway inflammation and remodeling, which has led to the hypothesis that aberrant activation of the asthmatic EMTU may lead to disease pathogenesis. Monoculture studies have aided in the understanding of the altered phenotype of airway epithelial and mesenchymal cells and their contribution to the pathogenesis of asthma. However, 3-dimensional (3D) co-culture models are needed to enable the study of epithelial-mesenchymal crosstalk in the setting of the in vivo environment. In this review, we summarize studies using 3D co-culture models to assess how defective epithelial-mesenchymal communication contributes to chronic airway inflammation and remodeling within the asthmatic EMTU.

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