scholarly journals In Vivo Vascularization of Anisotropic Channeled Porous Polylactide-Based Capsules for Islet Transplantation: The Effects of Scaffold Architecture and Implantation Site

2015 ◽  
pp. S75-S84 ◽  
Author(s):  
N. KASOJU ◽  
D. KUBIES ◽  
E. FÁBRYOVÁ ◽  
J. KŘÍŽ ◽  
M. M. KUMOREK ◽  
...  
Keyword(s):  
Islet Transplantation ◽  
Porous Scaffold ◽  
Oxygen Demand ◽  
Inflammatory Cells ◽  
Host Cells ◽  
Cell Infiltration ◽  
Greater Omentum ◽  
Brown Norway ◽  
Porous Scaffolds

The replacement of pancreatic islets for the possible treatment of type 1 diabetes is limited by the extremely high oxygen demand of the islets. To this end, here we hypothesize to create a novel extra-hepatic highly-vascularized bioartificial cavity using a porous scaffold as a template and using the host body as a living bioreactor for subsequent islet transplantation. Polylactide-based capsular-shaped anisotropic channeled porous scaffolds were prepared by following the unidirectional thermally-induced phase separation technique, and were implanted under the skin and in the greater omentum of Brown Norway rats. Polyamide mesh-based isotropic regular porous capsules were used as the controls. After 4weeks, the implants were excised and analyzed by histology. The hematoxylin and eosin, as well as Masson's trichrome staining, revealed a) low or no infiltration of giant inflammatory cells in the implant, b) minor but insignificant fibrosis around the implant, c) guided infiltration of host cells in the test capsule in contrast to random cell infiltration in the control capsule, and d) relatively superior cell infiltration in the capsules implanted in the greater omentum than in the capsules implanted under the skin. Furthermore, the anti-CD31 immunohistochemistry staining revealed numerous vessels at the implant site, but mostly on the external surface of the capsules. Taken together, the current study, the first of its kind, is a significant step-forward towards engineering a bioartificial microenvironment for the transplantation of islets.

scholarly journals The role of Leishmania GP63 in the modulation of innate inflammatory response to Leishmania major infection

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0262158
Author(s):  
Aretha Chan ◽  
Jose-Mauricio Ayala ◽  
Fernando Alvarez ◽  
Ciriaco Piccirillo ◽  
George Dong ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Leishmania Major ◽  
Critical Role ◽  
Protozoan Parasite ◽  
Inflammatory Cells ◽  
Host Cells ◽  
Cell Infection ◽  
Zinc Metalloprotease

Leishmaniasis is a disease caused by the protozoan parasite Leishmania and is known to affect millions of individuals worldwide. In recent years, we have established the critical role played by Leishmania zinc-metalloprotease GP63 in the modulation of host macrophage signalling and functions, favouring its survival and progression within its host. Leishmania major lacking GP63 was reported to cause limited infection in mice, however, it is still unclear how GP63 may influence the innate inflammatory response and parasite survival in an in vivo context. Therefore, we were interested in analyzing the early innate inflammatory events upon Leishmania inoculation within mice and establish whether Leishmania GP63 influences this initial inflammatory response. Experimentally, L. major WT (L. majorWT), L. major GP63 knockout (L. majorKO), or L. major GP63 rescue (L. majorR) were intraperitoneally inoculated in mice and the inflammatory cells recruited were characterized microscopically and by flow cytometry (number and cell type), and their infection determined. Pro-inflammatory markers such as cytokines, chemokines, and extracellular vesicles (EVs, e.g. exosomes) were monitored and proteomic analysis was performed on exosome contents. Data obtained from this study suggest that Leishmania GP63 does not significantly influence the pathogen-induced inflammatory cell recruitment, but rather their activation status and effector function. Concordantly, internalization of promastigotes during early infection could be influenced by GP63 as fewer L. majorKO amastigotes were found within host cells and appear to maintain in host cells over time. Collectively this study provides a clear analysis of innate inflammatory events occurring during L. major infection and further establish the prominent role of the virulence factor GP63 to provide favourable conditions for host cell infection.

scholarly journals Three dimensional porous scaffolds derived from collagen, elastin and fibrin proteins orchestrate adipose tissue regeneration

2021 ◽  
Vol 12 ◽  
pp. 204173142110192
Author(s):  
Prasad Sawadkar ◽  
Nandin Mandakhbayar ◽  
Kapil D Patel ◽  
Jennifer Olmas Buitrago ◽  
Tae Hyun Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Soft Tissue ◽  
Porous Scaffold ◽  
Pathological Condition ◽  
Donor Site ◽  
Porous Scaffolds ◽  
Adipose Derived Stem Cells ◽  
Natural Polymers

Current gold standard to treat soft tissue injuries caused by trauma and pathological condition are autografts and off the shelf fillers, but they have inherent weaknesses like donor site morbidity, immuno-compatibility and graft failure. To overcome these limitations, tissue-engineered polymers are seeded with stem cells to improve the potential to restore tissue function. However, their interaction with native tissue is poorly understood so far. To study these interactions and improve outcomes, we have fabricated scaffolds from natural polymers (collagen, fibrin and elastin) by custom-designed processes and their material properties such as surface morphology, swelling, wettability and chemical cross-linking ability were characterised. By using 3D scaffolds, we comprehensive assessed survival, proliferation and phenotype of adipose-derived stem cells in vitro. In vivo, scaffolds were seeded with adipose-derived stem cells and implanted in a rodent model, with X-ray microtomography, histology and immunohistochemistry as read-outs. Collagen-based materials showed higher cell adhesion and proliferation in vitro as well as higher adipogenic properties in vivo. In contrast, fibrin demonstrated poor cellular and adipogenesis properties but higher angiogenesis. Elastin formed the most porous scaffold, with cells displaying a non-aggregated morphology in vitro while in vivo elastin was the most degraded scaffold. These findings of how polymers present in the natural polymers mimicking ECM and seeded with stem cells affect adipogenesis in vitro and in vivo can open avenues to design 3D grafts for soft tissue repair.

scholarly journals 3D printed Ti6Al4V bone scaffolds with different pore structure effects on bone ingrowth

2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Fuyuan Deng ◽  
Linlin Liu ◽  
Zhong Li ◽  
Juncai Liu
Keyword(s):  
Bone Growth ◽  
Porous Scaffold ◽  
Bone Ingrowth ◽  
Fluid Velocity ◽  
Porous Titanium ◽  
Vital Role ◽  
Tissue Growth ◽  
Porous Scaffolds ◽  
Bone Scaffolds

AbstractThe microstructure of porous scaffolds plays a vital role in bone regeneration, but its optimal shape is still unclear. In this study, four kinds of porous titanium alloy scaffolds with similar porosities (65%) and pore sizes (650 μm) and different structures were prepared by selective laser melting. Four scaffolds were implanted into the distal femur of rabbits to evaluate bone tissue growth in vivo. Micro-CT and hard tissue section analyses were performed 6 and 12 weeks after the operation to reveal the bone growth of the porous scaffold. The results show that diamond lattice unit (DIA) bone growth is the best of the four topological scaffolds. Through computational fluid dynamics (CFD) analysis, the permeability, velocity and flow trajectory inside the scaffold structure were calculated. The internal fluid velocity difference of the DIA structure is the smallest, and the trajectory of fluid flow inside the scaffold is the longest, which is beneficial for blood vessel growth, nutrient transport and bone formation. In this study, the mechanism of bone growth in different structures was revealed by in vivo experiments combined with CFD, providing a new theoretical basis for the design of bone scaffolds in the future.

scholarly journals Examination of Early Interactions betweenHaemophilus ducreyi and Host Cells by Using Cocultured HaCaT Keratinocytes and Foreskin Fibroblasts

1999 ◽  
Vol 67 (10) ◽  
pp. 5352-5360 ◽  
Author(s):  
Franca R. Zaretzky ◽  
Thomas H. Kawula
Keyword(s):  
Hacat Cell ◽  
Inflammatory Cells ◽  
Etiologic Agent ◽  
Host Cells ◽  
Hacat Cells ◽  
Tissue Destruction ◽  
Necrosis Factor Alpha ◽  
Potent Inducer ◽  
Ulcer Disease

ABSTRACT Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease. Keratinocytes are likely the first cell type encountered by H. ducreyiupon infection of human skin; thus, the interaction between H. ducreyi and keratinocytes is probably important for the ability of H. ducreyi to establish infection. We have used the HaCaT keratinocyte cell line grown in monolayers and in cocultures with HS27 fibroblasts to investigate H. ducreyi interactions with keratinocytes and the host-cell response to H. ducreyiinfection. Using quantitative adherence and gentamicin protection assays, we determined that approximately 13% of H. ducreyiadhered to HaCaT cell monolayers, while only a small proportion (0.0052%) was intracellular. By transmission electron microscopy, we observed numerous H. ducreyi organisms adherent to but rarely within HaCaT cells cocultured with fibroblasts. Both liveH. ducreyi and purified H. ducreyilipooligosaccharide (LOS) induced significant interleukin 8 (IL-8) expression from HaCaT cell-HS27 cell cocultures. However, the level of IL-8 expression in response to LOS alone was not as pronounced.H. ducreyi LOS was a more potent inducer of IL-8 from cocultures than Escherichia coli lipopolysaccharide (LPS) at the same concentration, suggesting a unique effect of H. ducreyi LOS on cocultures. Neither live H. ducreyinor purified H. ducreyi LOS or E. coli LPS induced tumor necrosis factor alpha expression from cocultures.H. ducreyi induced drastically different cytokine profiles from cocultures than from HS27 or HaCaT cells cultured separately. IL-8 expression by skin cells in response to H. ducreyiinfection in vivo may be responsible for the massive influx of polymorphonuclear leukocytes and other inflammatory cells to the site of infection. This influx of inflammatory cells may be partly responsible for the tissue destruction characteristic of chancroid.

Fabrication and characterization of an acellular annulus fibrosus scaffold with aligned porous construct for tissue engineering

2021 ◽  
pp. 088532822110419
Author(s):  
Xiaoyu Jin ◽  
Ran Kang ◽  
Rongrong Deng ◽  
Xu Zhao ◽  
Zihan Wang ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Annulus Fibrosus ◽  
Freeze Drying ◽  
Porous Scaffold ◽  
Three Dimensional ◽  
Supporting Cell ◽  
Host Cells ◽  
Drying Methods ◽  
Freeze Dried

Scaffolds mimicking the native annulus fibrosus (AF) extracellular matrix (ECM) structure are crucial to guide the seeding cells to regenerate aligned tissue, while fabricating such a scaffold by synthetic material is challengeable. Native acellular scaffolds derived from AF tissue certainly possess the advantages of natural structure and composition. Based on previous studies, we modified decellularization procedure and especially compared two drying methods, including gradient dehydration and freeze-drying. The decellularization process can effectively remove the host cells and antigens such as α-Gal, while maintaining the original ECM including GAG and collagen I. Compared with gradient dehydration, freeze-drying not only rendered the decellularized scaffold in dry state for storage but also gave the scaffold more aligned porous structure and hydrophilicity. And, the acellular porous scaffold manifested better capacity of supporting cell ingrowth when seeded human bone marrow mesenchymal stem cells (hBMSCs) or implanted in vivo. Furthermore, this optimized freeze-dried scaffold showed similar mechanical elastic modulus as native AF and demonstrated rare inflammatory granuloma and immune rejection as observed in HE staining and immunohistochemistry staining (IHC) of CD8 and MAC387 epitopes when implanted subcutaneously in vivo. To sum up, through our decellularization and freeze-drying procedure, an aligned porous three-dimensional scaffold derived from the natural AF ECM was successfully fabricated with good retention of ECM components and benign biocompatibility. It will be a promising scaffold for AF tissue engineering.

On the Bio-Mechanical Properties of a Dual-Porous Osteochondral Scaffold

2008 ◽  
Author(s):  
Hajar Sharif ◽  
Yaser Shanjani ◽  
Mihaela Vlasea ◽  
Ehsan Toyserkani
Keyword(s):  
Mechanical Properties ◽  
Finite Element ◽  
Porous Scaffold ◽  
Element Model ◽  
Homogeneous Distribution ◽  
Porous Scaffolds ◽  
Osteochondral Scaffold ◽  
Apparent Stiffness

This work is concerned with the finite element modeling of a dual-porous scaffold including both fine and coarse pores. The layer with coarse pores is suitable for bone in vivo ingrowth and the finer pore layer is appropriate for in vitro cartilage culturing. Such scaffolds can be extensively used for repairing of osteochondral defects. The bio-mechanical properties of the proposed scaffold, including apparent stiffness and strain-based capability of the cell ingrowth, are identified using a 3D Finite Element Model. Moreover, to study the effect of the second layer on the strength of the whole scaffold, the stiffness of the dual and single-porous scaffolds was compared. The result of this study shows that the stiffness decreases by adding the second layer to a single-porous scaffold. Additionally, principal strain histograms of the single and the dual-porous scaffolds are compared to assess the effect of added layer on the capability for cell ingrowth stimulation of the whole structure. According to the results, the dual-porous scaffold provides more homogeneous distribution but a smaller amount of micro-strains which may cause different cell-growth behavior.

Development of a Hydroxyapatite Bone Tissue Engineering Scaffold with a Trimodal Pore Structure

2007 ◽  
Vol 361-363 ◽  
pp. 931-934 ◽  
Author(s):  
Conor T. Buckley ◽  
K.U. O’Kelly
Keyword(s):  
Tissue Engineering ◽  
Cell Viability ◽  
Porous Scaffold ◽  
Tissue Engineering Scaffold ◽  
Porous Scaffolds ◽  
Cell Penetration ◽  
Nutrient Delivery ◽  
And Migration

Tissue-engineering scaffold-based strategies have suffered from limited cell depth viability when cultured in vitro, with viable cells existing within the outer 250-500μm from the fluid-scaffold interface. This is primarily believed to be due to the lack of nutrient delivery into and waste removal from the inner regions of the scaffold construct. Other issues associated with porous scaffolds involve poor seeding efficiencies and limited cell penetration resulting in heterogeneous cellular distributions. This work focuses on the development a novel hydroxyapatite multi-domain porous scaffold architecture (i.e. a scaffold providing a discrete domain for cell occupancy and a separate domain for nutrient delivery) with the specific objectives of embodying in one scaffold the structures required to optimise cell seeding, cell proliferation and migration and potentially to facilitate vascularisation once implanted in vivo. This paper presents the development of the multidomain architecture and preliminary results on cell viability which show a significant improvement in cell viability in the scaffold interiors.

Healing effects of curcumin nanoparticles in deep tissue injury mouse model.

2020 ◽  
Vol 18 ◽  
Author(s):  
Zirui Zhang ◽  
Shangcong Han ◽  
Panpan Liu ◽  
Xu Yang ◽  
Jing Han ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mrna Expression ◽  
Tissue Injury ◽  
Inflammatory Cells ◽  
Pcr Analysis ◽  
Protective Effects ◽  
Deep Tissue ◽  
Deep Tissue Injury

Background: Chronic inflammation and lack of angiogenesis are the important pathological mechanisms in deep tissue injury (DTI). Curcumin is a well-known anti-inflammatory and antioxidant agent. However, curcumin is unstable under acidic and alkaline conditions, and can be rapidly metabolized and excreted in the bile, which shortens its bioactivity and efficacy. Objective: This study aimed to prepare curcumin-loaded poly (lactic-co-glycolic acid) nanoparticles (CPNPs) and to elucidate the protective effects and underlying mechanisms of wound healing in DTI models. Methods: CPNPs were evaluated for particle size, biocompatibility, in vitro drug release and their effect on in vivo wound healing. Results : The results of in vivo wound closure analysis revealed that CPNP treatments significantly improved wound contraction rates (p<0.01) at a faster rate than other three treatment groups. H&E staining revealed that CPNP treatments resulted in complete epithelialization and thick granulation tissue formation, whereas control groups resulted in a lack of compact epithelialization and persistence of inflammatory cells within the wound sites. Quantitative real-time PCR analysis showed that treatment with CPNPs suppressed IL-6 and TNF-α mRNA expression, and up-regulated TGF-β, VEGF-A and IL-10 mRNA expression. Western blot analysis showed up-regulated protein expression of TGF-β, VEGF-A and phosphorylatedSTAT3. Conclusion: Our results showed that CPNPs enhanced wound healing in DTI models, through modulation of the JAK2/STAT3 signalling pathway and subsequent upregulation of pro-healing factors.

ACIS, A Novel KepTide™, Binds to ACE-2 Receptor and Inhibits the Infection of SARS-CoV2 Virus in vitro in Primate Kidney Cells: Therapeutic Implications for COVID-19 (Preprint)

2020 ◽  
Author(s):  
Avik Sotira Scientific
Keyword(s):  
Social Life ◽  
Plaque Assay ◽  
Host Cells ◽  
Surface Glycoprotein ◽  
Crystallographic Structure ◽  
Therapeutic Implications ◽  
Biochemical Assays ◽  
Targeted Medicine

UNSTRUCTURED Coronavirus disease 2019 (COVID-19) is a severe acute respiratory syndrome (SARS) caused by a virus known as SARS-Coronavirus 2 (SARS-CoV2). Without a targeted-medicine, this disease has been causing a massive humanitarian crisis not only in terms of mortality, but also imposing a lasting damage to social life and economic progress of humankind. Therefore, an immediate therapeutic strategy needs to be intervened to mitigate this global crisis. Here, we report a novel KepTide™ (Knock-End Peptide) therapy that nullifies SARS-CoV2 infection. SARS-CoV2 employs its surface glycoprotein “spike” (S-glycoprotein) to interact with angiotensin converting enzyme-2 (ACE-2) receptor for its infection in host cells. Based on our in-silico-based homology modeling study validated with a recent X-ray crystallographic structure (PDB ID:6M0J), we have identified that a conserved motif of S-glycoprotein that intimately engages multiple hydrogen-bond (H-bond) interactions with ACE-2 enzyme. Accordingly, we designed a peptide, termed as ACIS (ACE-2 Inhibitory motif of Spike), that displayed significant affinity towards ACE-2 enzyme as confirmed by biochemical assays such as BLItz and fluorescence polarization assays. Interestingly, more than one biochemical modifications were adopted in ACIS in order to enhance the inhibitory action of ACIS and hence called as KEpTide™. Consequently, a monolayer invasion assay, plaque assay and dual immunofluorescence analysis further revealed that KEpTide™ efficiently mitigated the infection of SARS-CoV2 in vitro in VERO E6 cells. Finally, evaluating the relative abundance of ACIS in lungs and the potential side-effects in vivo in mice, our current study discovers a novel KepTide™ therapy that is safe, stable, and robust to attenuate the infection of SARS-CoV2 virus if administered intranasally. INTERNATIONAL REGISTERED REPORT RR2-https://doi.org/10.1101/2020.10.13.337584

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