scholarly journals In vivo cartilage regeneration in a multi-layered articular cartilage architecture mimicking scaffold

2020 ◽  
Vol 9 (9) ◽  
pp. 601-612
Author(s):  
Karthikeyan Rajagopal ◽  
Sowmya Ramesh ◽  
Noel Malcolm Walter ◽  
Aditya Arora ◽  
Dhirendra S. Katti ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Viability ◽  
Cartilage Regeneration ◽  
Fold Increase ◽  
Vital Role ◽  
Matrix Synthesis ◽  
Cartilage Formation ◽  
And Function

Aims Extracellular matrix (ECM) and its architecture have a vital role in articular cartilage (AC) structure and function. We hypothesized that a multi-layered chitosan-gelatin (CG) scaffold that resembles ECM, as well as native collagen architecture of AC, will achieve superior chondrogenesis and AC regeneration. We also compared its in vitro and in vivo outcomes with randomly aligned CG scaffold. Methods Rabbit bone marrow mesenchymal stem cells (MSCs) were differentiated into the chondrogenic lineage on scaffolds. Quality of in vitro regenerated cartilage was assessed by cell viability, growth, matrix synthesis, and differentiation. Bilateral osteochondral defects were created in 15 four-month-old male New Zealand white rabbits and segregated into three treatment groups with five in each. The groups were: 1) untreated and allogeneic chondrocytes; 2) multi-layered scaffold with and without cells; and 3) randomly aligned scaffold with and without cells. After four months of follow-up, the outcome was assessed using histology and immunostaining. Results In vitro testing showed that the secreted ECM oriented itself along the fibre in multi-layered scaffolds. Both types of CG scaffolds supported cell viability, growth, and matrix synthesis. In vitro chondrogenesis on scaffold showed an around 400-fold increase in collagen type 2 (COL2A1) expression in both CG scaffolds, but the total glycosaminoglycan (GAG)/DNA deposition was 1.39-fold higher in the multi-layered scaffold than the randomly aligned scaffold. In vivo cartilage formation occurred in both multi-layered and randomly aligned scaffolds treated with and without cells, and was shown to be of hyaline phenotype on immunostaining. The defects treated with multi-layered + cells, however, showed significantly thicker cartilage formation than the randomly aligned scaffold. Conclusion We demonstrated that MSCs loaded CG scaffold with multi-layered zonal architecture promoted superior hyaline AC regeneration. Cite this article: Bone Joint Res 2020;9(9):601–612.

Combined Effects of Bone Marrow-Derived Mesenchymal Stem Cells and PLGA-PEG-PLGA Scaffolds on Repair of Articular Cartilage Defect

2013 ◽  
Vol 815 ◽  
pp. 345-349 ◽  
Author(s):  
Ching Wen Hsu ◽  
Ping Liu ◽  
Song Song Zhu ◽  
Feng Deng ◽  
Bi Zhang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Articular Cartilage ◽  
Growth Factor ◽  
Cartilage Defect ◽  
Cartilage Regeneration ◽  
Tissue Growth ◽  
Cartilage Defects

Here we reported a combined technique for articular cartilage repair, consisting of bone arrow mesenchymal stem cells (BMMSCs) and poly (dl-lactide-co-glycolide-b-ethylene glycol-b-dl-lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymers carried with tissue growth factor (TGF-belat1). In the present study, BMMSCs seeded on PLGA-PEG-PLGA with were incubated in vitro, carried or not TGF-belta1, Then the effects of the composite on repair of cartilage defect were evaluated in rabbit knee joints in vivo. Full-thickness cartilage defects (diameter: 5 mm; depth: 3 mm) in the patellar groove were either left empty (n=18), implanted with BMMSCs/PLGA (n=18), TGF-belta1 modified BMMSCs/PLGA-PEG-PLGA. The defect area was examined grossly, histologically at 6, 24 weeks postoperatively. After implantation, the BMMSCs /PLGA-PEG-PLGA with TGF-belta1 group showed successful hyaline-like cartilage regeneration similar to normal cartilage, which was superior to the other groups using gross examination, qualitative and quantitative histology. These findings suggested that a combination of BMMSCs/PLGA-PEG-PLGA carried with tissue growth factor (TGF-belat1) may be an alternative treatment for large osteochondral defects in high loading sites.

FKBP51 Modulates Hippocampal Size and Function in Post-translational Regulation of Parkin

2021 ◽  
Author(s):  
Bin Qiu ◽  
Zhaohui Zhong ◽  
Shawn Righter ◽  
Yuxue Xu ◽  
Jun Wang ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Translational Regulation ◽  
Disease Onset ◽  
Fold Increase ◽  
Knock Out ◽  
Fk506 Binding Protein ◽  
Protein Levels ◽  
And Function

Abstract FK506-binding protein 51 (encoded by Fkpb51) has been associated with stress-related mental illness. To identify its function, we studied the morphological consequences of Fkbp51 deletion. Artificial Intelligence-assist morphological analysis identified that Fkbp51 knock-out (KO) mice possess more elongated CA and DG but shorter in height in coronal section when compared to WT. Primary cultured Fkbp51 KO hippocampal neurons were shown to exhibit larger dendritic outgrowth than wild-type (WT) controls, pharmacological manipulation experiments suggest that this may occur through regulation of microtubule-associated protein. Both in vitro primary culture and in vivo labeling support that FKBP51 regulates microtubule-associated protein expression. Furthermore, in the absence of differences in mRNA expression, Fkbp51 KO hippocampus exhibited decreases in βIII-tubulin, MAP2, and Tau protein levels, but a greater than 2.5-fold increase in Parkin protein. Overexpression and knock-down FKBP51 demonstrated that FKBP51 negatively regulates Parkin in a dose-dependent and ubiquitin-mediated manner. These results indicate a potential novel post-translational regulatory of Parkin by FKBP51 and significance of their interaction on disease onset.

scholarly journals Xenogenic Implantation of Equine Synovial Fluid-Derived Mesenchymal Stem Cells Leads to Articular Cartilage Regeneration

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Mohammed Zayed ◽  
Steven Newby ◽  
Nabil Misk ◽  
Robert Donnell ◽  
Madhu Dhar
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Articular Cartilage ◽  
Synovial Fluid ◽  
Cartilage Repair ◽  
Cartilage Regeneration ◽  
Large Animal

Horses are widely used as large animal preclinical models for cartilage repair studies, and hence, there is an interest in using equine synovial fluid-derived mesenchymal stem cells (SFMSCs) in research and clinical applications. Since, we have previously reported that similar to bone marrow-derived MSCs (BMMSCs), SFMSCs may also exhibit donor-to-donor variations in their stem cell properties; the current study was carried out as a proof-of-concept study, to compare the in vivo potential of equine BMMSCs and SFMSCs in articular cartilage repair. MSCs from these two sources were isolated from the same equine donor. In vitro analyses confirmed a significant increase in COMP expression in SFMSCs at day 14. The cells were then encapsulated in neutral agarose scaffold constructs and were implanted into two mm diameter full-thickness articular cartilage defect in trochlear grooves of the rat femur. MSCs were fluorescently labeled, and one week after treatment, the knee joints were evaluated for the presence of MSCs to the injured site and at 12 weeks were evaluated macroscopically, histologically, and then by immunofluorescence for healing of the defect. The macroscopic and histological evaluations showed better healing of the articular cartilage in the MSCs’ treated knee than in the control. Interestingly, SFMSC-treated knees showed a significantly higher Col II expression, suggesting the presence of hyaline cartilage in the healed defect. Data suggests that equine SFMSCs may be a viable option for treating osteochondral defects; however, their stem cell properties require prior testing before application.

scholarly journals Effect of different aged cartilage ECM on chondrogenesis of BMSCs in vitro and in vivo

2020 ◽  
Vol 7 (6) ◽  
pp. 583-595
Author(s):  
Xiuyu Wang ◽  
Yan Lu ◽  
Wan Wang ◽  
Qiguang Wang ◽  
Jie Liang ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Current Knowledge ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Biological Properties ◽  
Cartilage Tissue ◽  
Before And After ◽  
Marrow Mesenchymal Stem Cells

Abstract Extracellular matrix (ECM)-based biomaterials are promising candidates in cartilage tissue engineering by simulating the native microenvironment to regulate the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) without exogenous growth factors. The biological properties of ECM scaffolds are primarily depended on the original source, which would directly influence the chondrogenic effects of the ECM materials. Despite the expanding investigations on ECM scaffolds in recent years, the selection of optimized ECM materials in cartilage regeneration was less reported. In this study, we harvested and compared the articular cartilage ECM from newborn, juvenile and adult rabbits. The results demonstrated the significant differences in the mechanical strength, sulphated glycosaminoglycan and collagen contents of the different aged ECM, before and after decellularization. Consequently, different compositional and mechanical properties were shown in the three ECM-based collagen hydrogels, which exerted age-dependent chondrogenic inducibility. In general, both in vitro and in vivo results suggested that the newborn ECM promoted the most chondrogenesis of BMSCs but led to severe matrix calcification. In contrast, BMSCs synthesized the lowest amount of cartilaginous matrix with minimal calcification with adult ECM. The juvenile ECM achieved the best overall results in promoting chondrogenesis of BMSCs and preventing matrix calcification. Together, this study provides important information to our current knowledge in the design of future ECM-based biomaterials towards a successful repair of articular cartilage.

Biomechanical issues of tissue-engineered constructs for articular cartilage regeneration: in vitro and in vivo approaches

2019 ◽  
Vol 132 (1) ◽  
pp. 53-80 ◽  
Author(s):  
Lucio Cipollaro ◽  
Maria Camilla Ciardulli ◽  
Giovanna Della Porta ◽  
Giuseppe M Peretti ◽  
Nicola Maffulli
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cartilage Regeneration ◽  
Biological Properties ◽  
Trophic Factors ◽  
Gene Therapies ◽  
Zonal Organization ◽  
Meta Analyses

Abstract Background Given the limited regenerative capacity of injured articular cartilage, the absence of suitable therapeutic options has encouraged tissue-engineering approaches for its regeneration or replacement. Sources of data Published articles in any language identified in PubMed and Scopus electronic databases up to August 2019 about the in vitro and in vivo properties of cartilage engineered constructs. A total of 64 articles were included following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Areas of agreement Regenerated cartilage lacks the biomechanical and biological properties of native articular cartilage. Areas of controversy There are many different approaches about the development of the architecture and the composition of the scaffolds. Growing points Novel tissue engineering strategies focus on the development of cartilaginous biomimetic materials able to repair cartilage lesions in association to cell, trophic factors and gene therapies. Areas timely for developing research A multi-layer design and a zonal organization of the constructs may lead to achieve cartilage regeneration.

scholarly journals Interstitial flow upregulates matrix synthesis and attenuates NF-kB signaling in a novel perfusion bioreactor for articular cartilage tissue engineering

2021 ◽  
Author(s):  
Haneen Abusharkh ◽  
Terreill Robertson ◽  
Juana Mendenhall ◽  
Bulent Gozen ◽  
Edwin Tingstad ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Fold Increase ◽  
Cartilage Tissue ◽  
Type I ◽  
Interstitial Flow ◽  
Perfusion System ◽  
Matrix Synthesis ◽  
Porous Chitosan

The present study is focused on designing an easy-to-use novel perfusion system for articular cartilage (AC) tissue engineering and using it to elucidate the mechanism by which interstitial shear upregulates matrix synthesis by articular chondrocytes (AChs). Porous chitosan-agarose (CHAG) scaffolds were synthesized, freeze-dried, and compared to bulk agarose (AG) scaffolds. Both scaffold types were seeded with osteoarthritic human AChs and cultured in a novel perfusion system for one week with a shear-inducing medium flow velocity of 0.33 mm/s corresponding to an average surficial shear of 0.4 mPa and a CHAG interstitial shear of 40 mPa. While there were no statistical differences in cell viability for perfusion vs. static cultures for either scaffold type, CHAG scaffold cultures exhibited 3.3-fold higher (p<0.005) cell viability compared to AG scaffold cultures. Effects of combined superficial and interstitial perfusion for CHAG showed 150- and 45-fold (p<0.0001) increases in total collagen (COL) and 13- and 2.2-fold (p<0.001) increases in glycosaminoglycans (GAGs) over AG’s scaffold non-perfusion and perfusion cultures, respectively, and a 1.5-fold and 3.6-fold (p<0.005) increase over non-perfusion CHAG cultures. Contrasting CHAG perfusion and static cultures, chondrogenic gene comparisons showed a 3.5-fold increase in collagen type II/type I (COL2A1/COL1A1) mRNA ratio (p<0.05), and a 1.3-fold increase in aggrecan mRNA. Observed effects are suggested to be the result of inhibiting the inflammatory NF-κB signal transduction pathway as confirmed by a further study that indicated a reduction by 3.2-fold (p<0.05) upon exposure to perfusion. Our results demonstrate that the presence of pores plays a critical role in improving cell viability and that interstitial flow caused by medium perfusion through the porous scaffolds enhances the expression of chondrogenic genes and ECM components through the downregulation of NF-κB1.

Bruton's Tyrosine Kinase Inhibitor PCI-32765 Abrogates BCR- and Nurselike Cell-Derived Activation of CLL Cells In Vitro and In Vivo.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 45-45 ◽  
Author(s):  
Sabine Ponader ◽  
Joseph Buggy ◽  
Susan O'Brien ◽  
William G. Wierda ◽  
Michael Keating ◽  
...  
Keyword(s):  
Cell Viability ◽  
Blood Plasma ◽  
B Cell ◽  
Clinical Activity ◽  
Surrogate Markers ◽  
Pre Treatment ◽  
And Function ◽  
Cell Chemotaxis

Abstract Abstract 45 Bruton's tyrosine kinase (Btk) plays a crucial role in development and function of normal B cells. Mutations in the gene encoding Btk cause a B cell defect, which manifests in boys during early childhood as X-linked agammaglobulinemia, a primary immunodeficiency originally described by Bruton in 1952. In normal B cells, Btk is involved in B-cell antigen receptor (BCR) signaling, but its function in chronic lymphocytic leukemia (CLL) cells has not yet been explored. PCI-32765 is a potent (IC50=0.5 nM), selective, and irreversible Btk inhibitor, which binds to the Cys-481 residue of Btk, causing an irreversible inhibition of Btk's enzymatic activity (Honigberg et al, Proc Natl Acad Sci USA 107(29):13075-80). The clinical activity of PCI-32765 is currently under investigation in patients with B cell malignancies, and preliminary results indicated significant clinical activity in CLL patients (Advani R. et al., ASCO 2010 meeting,JCO 28: 8012, 2010). We investigated the effect of PCI-32765 (conc. 1 mM) on CLL cell viability and activation after BCR triggering with anti-IgM and in co-cultures with nurselike cells (NLC). After 24h incubation with PCI-32765, the viability of anti-IgM-stimulated CLL cells decreased significantly to 79.4% ± 4.6% of respective controls (n=10). In NLC co-cultures, CLL cell viability also was significantly decreased in the presence of PCI-32765. For example, after 24h incubation with PCI-32765, the mean viability of CLL cells was 78.7 ± 5.1% compared to respective controls (see Fig. A). The chemokines CCL3 and CCL4 are secreted by CLL cells after BCR triggering and in NLC co-cultures, and function as surrogate markers of BCR-derived activation of CLL cells (Burger JA et al., Blood 113:3050-8, 2009). Therefore, we measured the effect of Btk inhibition by PCI-32765 on secretion of these chemokines in vitro and in vivo. In CLL cells co-cultured with NLC, the secretion of these chemokines was significantly reduced after 24 hours incubation with 1 mM PCI-32765 from 393 ± 172 pg/mL to 54 ± 46 pg/mL (CCL3) or from 2550 ± 678 pg/mL to 394 ± 188 pg/mL (CCL4, mean ± SD, n=11). Plasma samples from CLL patients treated with PCI-32765 (ongoing Phase 1 study) revealed high pre-treatment CCL3/4 levels, and these levels were significantly decreased after treatment. For example, 24 hours following the first dose of PCI-32765, CCL3 levels decreased from 60 ± 29 pg/mL to 16 ± 13 pg/mL, and CCL4 pre-treatment levels decreased from 106 ± 55 pg/mL to 23 ± 12 pg/mL (mean ± SD, n=6, see Fig. B). The clinical activity of PCI-32765 included a rapid (24 hour) reduction in lymphadenopathy accompanied by a transient lymphocytosis, suggesting that the drug might affect cell homing or migration to factors in tissue microenvironments. So we examined the impact of PCI-32765 on CLL cell chemotaxis towards the chemokines CXCL12 and CXCL13. Chemotaxis was significantly reduced by 1 mM PCI-32765 to levels that were 57 ± 9% (CXCL12) or 46 ± 5% (CXCL13) of respective controls (mean ± SD, n=10). This data is consistent with a compartmental shift of CLL cells from tissues to the blood, explaining the lymphocytosis early during treatment with PCI-32765. Collectively, our data demonstrate that PCI-32765 is an effective inhibitor of BCR- and NLC-derived survival signals, and interferes with CLL cell chemotaxis. PCI-32765 rapidly downregulates BCR-derived CLL cell activation in vivo, using CCL3/4 plasma levels as surrogate markers. These studies provide insight into the activity of PCI-32765 in CLL and help explaining the fascinating clinical activity of this new agent. Figure (A) CLL cell viability in suspension and nurselike (NLC) co-culture after 24 hours incubation with 1mM PCI-32765, n=7 and (B) CCL3 concentrations in blood plasma of CLL patients after treatment with PCI-32765 (n=6, Mean+/−SEM). Figure. (A) CLL cell viability in suspension and nurselike (NLC) co-culture after 24 hours incubation with 1mM PCI-32765, n=7 and (B) CCL3 concentrations in blood plasma of CLL patients after treatment with PCI-32765 (n=6, Mean+/−SEM). Disclosures: Buggy: Pharmacyclics, Inc.: Employment.

scholarly journals Mesenchymal Stem Cells in Oriented PLGA/ACECM Composite Scaffolds Enhance Structure-Specific Regeneration of Hyaline Cartilage in a Rabbit Model

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Weimin Guo ◽  
Xifu Zheng ◽  
Weiguo Zhang ◽  
Mingxue Chen ◽  
Zhenyong Wang ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Hyaline Cartilage ◽  
Rabbit Model ◽  
Cartilage Regeneration ◽  
Cartilage Defects ◽  
Composite Scaffolds ◽  
Human Cartilage ◽  
Native Cartilage

Articular cartilage lacks a blood supply and nerves. Hence, articular cartilage regeneration remains a major challenge in orthopedics. Decellularized extracellular matrix- (ECM-) based strategies have recently received particular attention. The structure of native cartilage exhibits complex zonal heterogeneity. Specifically, the development of a tissue-engineered scaffold mimicking the aligned structure of native cartilage would be of great utility in terms of cartilage regeneration. Previously, we fabricated oriented PLGA/ACECM (natural, nanofibrous, articular cartilage ECM) composite scaffolds. In vitro, we found that the scaffolds not only guided seeded cells to proliferate in an aligned manner but also exhibited high biomechanical strength. To detect whether oriented cartilage regeneration was possible in vivo, we used mesenchymal stem cell (MSC)/scaffold constructs to repair cartilage defects. The results showed that cartilage defects could be completely regenerated. Histologically, these became filled with hyaline cartilage and subchondral bone. Moreover, the aligned structure of cartilage was regenerated and was similar to that of native tissue. In conclusion, the MSC/scaffold constructs enhanced the structure-specific regeneration of hyaline cartilage in a rabbit model and may be a promising treatment strategy for the repair of human cartilage defects.

scholarly journals In Vivo Articular Cartilage Regeneration Using Human Dental Pulp Stem Cells Cultured in an Alginate Scaffold: A Preliminary Study

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Manuel Mata ◽  
Lara Milian ◽  
Maria Oliver ◽  
Javier Zurriaga ◽  
Maria Sancho-Tello ◽  
...  
Keyword(s):  
Stem Cells ◽  
Articular Cartilage ◽  
Dental Pulp ◽  
Cartilage Regeneration ◽  
Cell Types ◽  
Dental Pulp Stem Cells ◽  
The Poor ◽  
Human Dental Pulp

Osteoarthritis is an inflammatory disease in which all joint-related elements, articular cartilage in particular, are affected. The poor regeneration capacity of this tissue together with the lack of pharmacological treatment has led to the development of regenerative medicine methodologies including microfracture and autologous chondrocyte implantation (ACI). The effectiveness of ACI has been shown in vitro and in vivo, but the use of other cell types, including bone marrow and adipose-derived mesenchymal stem cells, is necessary because of the poor proliferation rate of isolated articular chondrocytes. In this investigation, we assessed the chondrogenic ability of human dental pulp stem cells (hDPSCs) to regenerate cartilage in vitro and in vivo. hDPSCs and primary isolated rabbit chondrocytes were cultured in chondrogenic culture medium and found to express collagen II and aggrecan. Both cell types were cultured in 3% alginate hydrogels and implanted in a rabbit model of cartilage damage. Three months after surgery, significant cartilage regeneration was observed, particularly in the animals implanted with hDPSCs. Although the results presented here are preliminary, they suggest that hDPSCs may be useful for regeneration of articular cartilage.

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