Human adipose-derived stromal/stem cells expressing doublecortin improve cartilage repair in rabbits and monkeys

AbstractLocalized cartilage lesions in early osteoarthritis and acute joint injuries are usually treated surgically to restore function and relieve pain. However, a persistent clinical challenge remains in how to repair the cartilage lesions. We expressed doublecortin (DCX) in human adipose-derived stromal/stem cells (hASCs) and engineered hASCs into cartilage tissues using an in vitro 96-well pellet culture system. The cartilage tissue constructs with and without DCX expression were implanted in the knee cartilage defects of rabbits (n = 42) and monkeys (n = 12). Cohorts of animals were euthanized at 6, 12, and 24 months after surgery to evaluate the cartilage repair outcomes. We found that DCX expression in hASCs increased expression of growth differentiation factor 5 (GDF5) and matrilin 2 in the engineered cartilage tissues. The cartilage tissues with DCX expression significantly enhanced cartilage repair as assessed macroscopically and histologically at 6, 12, and 24 months after implantation in the rabbits and 24 months after implantation in the monkeys, compared to the cartilage tissues without DCX expression. These findings suggest that hASCs expressing DCX may be engineered into cartilage tissues that can be used to treat localized cartilage lesions.

Download Full-text

Stem Cells for Cartilage Repair: Preclinical Studies and Insights in Translational Animal Models and Outcome Measures

Stem Cells International ◽

10.1155/2018/9079538 ◽

2018 ◽

Vol 2018 ◽

pp. 1-22 ◽

Cited By ~ 28

Author(s):

Melissa Lo Monaco ◽

Greet Merckx ◽

Jessica Ratajczak ◽

Pascal Gervois ◽

Petra Hilkens ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Animal Models ◽

Outcome Measures ◽

Cartilage Repair ◽

Cartilage Tissue ◽

Preclinical Studies ◽

Paracrine Factors ◽

Advantages And Disadvantages

Due to the restricted intrinsic capacity of resident chondrocytes to regenerate the lost cartilage postinjury, stem cell-based therapies have been proposed as a novel therapeutic approach for cartilage repair. Moreover, stem cell-based therapies using mesenchymal stem cells (MSCs) or induced pluripotent stem cells (iPSCs) have been used successfully in preclinical and clinical settings. Despite these promising reports, the exact mechanisms underlying stem cell-mediated cartilage repair remain uncertain. Stem cells can contribute to cartilage repair via chondrogenic differentiation, via immunomodulation, or by the production of paracrine factors and extracellular vesicles. But before novel cell-based therapies for cartilage repair can be introduced into the clinic, rigorous testing in preclinical animal models is required. Preclinical models used in regenerative cartilage studies include murine, lapine, caprine, ovine, porcine, canine, and equine models, each associated with its specific advantages and limitations. This review presents a summary of recentin vitrodata and fromin vivopreclinical studies justifying the use of MSCs and iPSCs in cartilage tissue engineering. Moreover, the advantages and disadvantages of utilizing small and large animals will be discussed, while also describing suitable outcome measures for evaluating cartilage repair.

Download Full-text

Engineered cartilage tissue from biodegradable Poly(ε-caprolactone) scaffold and human umbilical cord derived mesenchymal stem cells

Biomedical Research and Therapy ◽

10.15419/bmrat.v5i02.414 ◽

2018 ◽

Vol 5 (02) ◽

pp. 2000-2012

Author(s):

Phuc Dang-Ngoc Nguyen ◽

Ngoc Bich Vu ◽

Ha Thi-Ngan Le ◽

Thuy Thi-Thanh Dao ◽

Long Xuan Gia ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Alcian Blue ◽

Cartilage Tissue ◽

Cartilage Defects ◽

Human Umbilical Cord ◽

Biodegradable Poly ◽

Safranin O ◽

Engineered Cartilage

Introduction: Cartilage injury is the most common injury among orthopedic diseases. The predominant treatment for this condition is cartilage transplantation. Therefore, production of cartilage for treatment is an important strategy in regenerative medicine of cartilage to provide surgeons with an additional option for treatment of cartilage defects. This study aimed to produce in vitro engineered cartilage tissue by culturing and differentiating umbilical cord derived mesenchymal stem cells on biodegradable Poly(ε-caprolactone) (PCL) scaffold. Methods: Human umbilical cord derived mesenchymal stem cells (UCMSCs) were isolated and expanded according to previous published protocols. UCMSCs were labeled with CD90 APC‑conjugated monoclonal antibody (CD90-APC) and then seeded onto porous PCL scaffolds. Cell adhesion and proliferation on PCL scaffolds were evaluated based on the strength/signal of APC, MTT assays, and scanning electron microscopy (SEM). The chondrogenic differentiation of UCMSCs on scaffolds was detected by Alcian Blue and Safranin O staining. Results: The results showed that UCMSCs successfully adhered, proliferated and differentiated into chondroblasts and chondrocytes on PCL scaffolds. The chondrocyte scaffolds were positive for some markers of cartilage, as indicated by Alcian Blue and Safranin O staining. Conclusion: In conclusion, this study showed successful production of cartilage tissues from UCMSCs on PCL scaffolds.

Download Full-text

Effects of chondrogenic and osteogenic regulatory factors on composite constructs grown using human mesenchymal stem cells, silk scaffolds and bioreactors

Journal of The Royal Society Interface ◽

10.1098/rsif.2007.1302 ◽

2008 ◽

Vol 5 (25) ◽

pp. 929-939 ◽

Cited By ~ 48

Author(s):

Alexander Augst ◽

Darja Marolt ◽

Lisa E Freed ◽

Charu Vepari ◽

Lorenz Meinel ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Formation ◽

Human Mesenchymal Stem Cells ◽

Tissue Formation ◽

Tissue Constructs ◽

Engineer Cartilage ◽

Silk Scaffolds ◽

Engineered Cartilage

Human mesenchymal stem cells (hMSCs) isolated from bone marrow aspirates were cultured on silk scaffolds in rotating bioreactors for three weeks with either chondrogenic or osteogenic medium supplements to engineer cartilage- or bone-like tissue constructs. Osteochondral composites formed from these cartilage and bone constructs were cultured for an additional three weeks in culture medium that was supplemented with chondrogenic factors, supplemented with osteogenic factors or unsupplemented. Progression of cartilage and bone formation and the integration between the two regions were assessed by medical imaging (magnetic resonance imaging and micro-computerized tomography imaging), and by biochemical, histological and mechanical assays. During composite culture (three to six weeks), bone-like tissue formation progressed in all three media to a markedly larger extent than cartilage-like tissue formation. The integration of the constructs was most enhanced in composites cultured in chondrogenic medium. The results suggest that tissue composites with well-mineralized regions and substantially less developed cartilage regions can be generated in vitro by culturing hMSCs on silk scaffolds in bioreactors, that hMSCs have markedly higher capacity for producing engineered bone than engineered cartilage, and that chondrogenic factors play major roles at early stages of bone formation by hMSCs and in the integration of the two tissue constructs into a tissue composite.

Download Full-text

Novel Biologically Inspired Nanostructured Scaffolds for Directing Chondrogenic Differentiation of Mesenchymal Stem Cells

MRS Proceedings ◽

10.1557/opl.2013.181 ◽

2013 ◽

Vol 1498 ◽

pp. 59-66 ◽

Cited By ~ 1

Author(s):

Benjamin Holmes ◽

Nathan J. Castro ◽

Jian Li ◽

Lijie Grace Zhang

Keyword(s):

Carbon Nanotubes ◽

Tissue Engineering ◽

Stem Cells ◽

Cartilage Tissue Engineering ◽

Modern Medicine ◽

Cartilage Tissue ◽

Cartilage Defects ◽

Medical Problems ◽

Biologically Inspired

ABSTRACTCartilage defects, which are caused by a variety of reasons such as traumatic injuries, osteoarthritis, or osteoporosis, represent common and severe clinical problems. Each year, over 6 million people visit hospitals in the U.S. for various knee, wrist, and ankle problems. As modern medicine advances, new and novel methodologies have been explored and developed in order to solve and improve current medical problems. One of the areas of investigation is tissue engineering [1, 2]. Since cartilage matrix is nanocomposite, the goal of the current work is to use nanomaterials and nanofabrication methods to create novel biologically inspired tissue engineered cartilage scaffolds for facilitating human bone marrow mesenchymal stem cell (MSC) chondrogenesis. For this purpose, through electrospinning techniques, we designed a series of novel 3D biomimetic nanostructured scaffolds based on carbon nanotubes and biocompatible poly(L-lactic acid) (PLLA) polymers. Specifically, a series of electrospun fibrous PLLA scaffolds with controlled fiber dimension and surface nanoporosity were fabricated in this study. In vitro hMSC studies showed that stem cells prefer to attach in the scaffolds with smaller fiber diameter or suitable nanoporous structures. More importantly, our in vitro differentiation results demonstrated that incorporation of the biomimetic carbon nanotubes and poly L-lysine coating can induce GAG and collagen synthesis that is indicative of chondrogenic differentiations of MSCs. Our novel scaffolds also performed better than controls, which make them promising for cartilage tissue engineering applications.

Download Full-text

Repair of Osteochondral Defects With Predifferentiated Mesenchymal Stem Cells of Distinct Phenotypic Character Derived From a Nanotopographic Platform

The American Journal of Sports Medicine ◽

10.1177/0363546520907137 ◽

2020 ◽

Vol 48 (7) ◽

pp. 1735-1747

Author(s):

Yingnan Wu ◽

Zheng Yang ◽

Vinitha Denslin ◽

XiaFei Ren ◽

Chang Sheng Lee ◽

...

Keyword(s):

Mechanical Properties ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Articular Cartilage ◽

Cartilage Repair ◽

Cartilage Regeneration ◽

Cartilage Tissue ◽

Cartilage Defects ◽

Defect Site

Background: Articular cartilage has a zonal architecture and biphasic mechanical properties. The recapitulation of surface lubrication properties with high compressibility of the deeper layers of articular cartilage during regeneration is essential in achieving long-term cartilage integrity. Current clinical approaches for cartilage repair, especially with the use of mesenchymal stem cells (MSCs), have yet to restore the hierarchically organized architecture of articular cartilage. Hypothesis: MSCs predifferentiated on surfaces with specific nanotopographic patterns can provide phenotypically stable and defined chondrogenic cells and, when delivered as a bilayered stratified construct at the cartilage defect site, will facilitate the formation of functionally superior cartilage tissue in vivo. Study Design: Controlled laboratory study. Methods: MSCs were subjected to chondrogenic differentiation on specific nanopatterned surfaces. The phenotype of the differentiated cells was assessed by the expression of cartilage markers. The ability of the 2-dimensional nanopattern-generated chondrogenic cells to retain their phenotypic characteristics after removal from the patterned surface was tested by subjecting the enzymatically harvested cells to 3-dimensional fibrin hydrogel culture. The in vivo efficacy in cartilage repair was demonstrated in an osteochondral rabbit defect model. Repair by bilayered construct with specific nanopattern predifferentiated cells was compared with implantation with cell-free fibrin hydrogel, undifferentiated MSCs, and mixed-phenotype nanopattern predifferentiated MSCs. Cartilage repair was evaluated at 12 weeks after implantation. Results: Three weeks of predifferentiation on 2-dimensional nanotopographic patterns was able to generate phenotypically stable chondrogenic cells. Implantation of nanopatterned differentiated MSCs as stratified bilayered hydrogel constructs improved the repair quality of cartilage defects, as indicated by histological scoring, mechanical properties, and polarized microscopy analysis. Conclusion: Our results indicate that with an appropriate period of differentiation, 2-dimensional nanotopographic patterns can be employed to generate phenotypically stable chondrogenic cells, which, when implanted as stratified bilayered hydrogel constructs, were able to form functionally superior cartilage tissue. Clinical Relevance: Our approach provides a relatively straightforward method of obtaining large quantities of zone-specific chondrocytes from MSCs to engineer a stratified cartilage construct that could recapitulate the zonal architecture of hyaline cartilage, and it represents a significant improvement in current MSC-based cartilage regeneration.

Download Full-text

Effect of intra-articular injection of adipose stem cells on traumatic osteoarthritis cartilage defects

Materials Express ◽

10.1166/mex.2021.1874 ◽

2021 ◽

Vol 11 (1) ◽

pp. 28-37

Author(s):

Qian Wang ◽

Na Yang ◽

Kun Zhang ◽

Zhong Li ◽

Yangjun Zhu ◽

...

Keyword(s):

Stem Cells ◽

Cartilage Repair ◽

Cartilage Defect ◽

Osteoblastic Differentiation ◽

Left Knee ◽

Inflammatory Factors ◽

Cartilage Defects ◽

Osteoarthritis Cartilage

Traumatic osteoarthritis with cartilage defects can lead to mobility problems. Mitotic activity in cartilage is extremely low, and once damaged, repairing can be difficult. The commonly used autologous or allogeneic cartilage transplantation techniques also have certain limitations. In recent years, directed induction of osteoblastic differentiation using adipocytes has been shown to be effective in repairing cartilage defects. However, it is often induced in vitro and is prone to incomplete or over-differentiation. In addition, because of the large differences in the in vivo and in vitro microenvironment, exploring the influence of these differences in the in vivo microenvironment on the directional differentiation of adipose-derived stem cells (ADSCs) and their effect on cartilage repair is necessary. In this study, a cartilage defect model in rabbits with traumatic osteoarthritis of the left knee was established, and different interventions were conducted in different groups. We determined the effect of directly injecting ADSCs into the joints on repairing cartilage defects in rabbits with traumatic osteoarthritis and analyzed the differences in repair time of newly developed cartilage defects and old cartilage frontal defects. The results indicated that the placement of a stent and injection of ADSCs improved the knee joint activity, increased the expression of BMP and TGF-β protein, and reduced the expression of inflammatory factors, including IL-1β, IL-6, IL-17, and TNF-α. No difference was found between the new cartilage defect and the old one. By directly observing the cartilage defect, intervention with ADSCs + scaffold increased the connection between the cartilage defect and the normal tissue and improved the cartilage repair effect. These results indicated that directly injecting ADSCs into the joints is an effective approach for repairing cartilage defects in traumatic osteoarthritis, and it was not affected by the age of the defect.

Download Full-text

Aptamer-Functionalized Bioscaffold Enhances Cartilage Repair by Improving Stem Cell Recruitment in Osteochondral Defects of Rabbit Knees

The American Journal of Sports Medicine ◽

10.1177/0363546519856355 ◽

2019 ◽

Vol 47 (10) ◽

pp. 2316-2326 ◽

Cited By ~ 11

Author(s):

Xin Wang ◽

Xiongbo Song ◽

Tao Li ◽

Jiajia Chen ◽

Guotao Cheng ◽

...

Keyword(s):

Stem Cells ◽

Silk Fibroin ◽

Cartilage Repair ◽

Osteochondral Defect ◽

Cartilage Tissue ◽

Control Group ◽

Osteochondral Defects ◽

Scaffold Group

Background: Recruitment of endogenous stem cells has been considered an alternative to cell injection/implantation in articular cartilage repair. Purpose: (1) To develop a cartilage tissue-engineering scaffold with clinically available biomaterials and functionalize the scaffold with an aptamer (Apt19s) that specifically recognizes pluripotent stem cells. (2) To determine whether this scaffold could recruit joint-resident mesenchymal stem cells (MSCs) when implanted into an osteochondral defect in a rabbit model and to examine the effects of cartilage regeneration. Study Design: Controlled laboratory study. Methods: The reinforced scaffold was fabricated by embedding a silk fibroin sponge into silk fibroin/hyaluronic acid–tyramine hydrogel and characterized in vitro. A cylindrical osteochondral defect (3.2 mm wide × 4 mm deep) was created in the trochlear grooves of rabbit knees. The rabbits were randomly assigned into 3 groups: Apt19s-functionalized scaffold group, scaffold-only group, and control group. Animals were sacrificed at 6 and 12 weeks after transplantation. Repaired tissues were evaluated via gross examination, histologic examination, and immunohistochemistry. Results: In vitro, this aptamer-functionalized scaffold could recruit bone marrow–derived MSCs and support cell adhesion. In vivo, the aptamer-functionalized scaffold enhanced cell homing in comparison with the aptamer-free scaffold. The aptamer-functionalized scaffold group also exhibited superior cartilage restoration when compared with the scaffold-only group and the control group. Conclusion: The Apt19s-functionalized scaffold exhibited the ability to recruit MSCs both in vitro and in vivo and achieved a better outcome of cartilage repair than the scaffold only or control in an osteochondral defect model. Clinical Relevance: The findings demonstrate a promising strategy of using aptamer-functionalized bioscaffolds for restoration of chondral/osteochondral defects via aptamer-introduced homing of MSCs.

Download Full-text

Comparison of chondrogenesis-related biological behaviors between human urine-derived stem cells and human bone marrow mesenchymal stem cells from the same individual

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02370-1 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Jiachen Sun ◽

Fei Xing ◽

Min Zou ◽

Min Gong ◽

Lang Li ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Bone Marrow ◽

Cartilage Repair ◽

Cartilage Defects ◽

In Vitro Experiments ◽

Rabbit Knee ◽

Marrow Mesenchymal Stem Cells

Abstract Background Stem cells are the main choice for seed cells in tissue engineering, but using most traditional stem cells requires invasive and complicated procedures. Human urine-derived stem cells (hUSCs) are an alternative stem cell source with the advantages of being isolated noninvasively and repetitively from the same individual. The aim of this study was to compare chondrogenesis-related biological behaviors between hUSCs and human bone marrow mesenchymal stem cells (hBMSCs) from the same individual. Methods hUSCs and hBMSCs were isolated from six patients who underwent iliac bone grafting. Cell morphology, proliferation, colony-forming, migration, and multidifferentiation analyses were performed in vitro. Then, acellular cartilage extracellular matrix (ACM) scaffolds were fabricated for in vivo implantation. The comparisons of cell viability, morphology, proliferation, and chondrogenesis between hUSCs and hBMSCs cultured on scaffolds were performed before implantation. The scaffolds loaded with hUSCs or hBMSCs were implanted into a rabbit knee model to repair cartilage defects. Magnetic resonance imaging (MRI) and micro-computed tomography (μCT) Analyses, inflammation and toxicity assays, gross observation, and histological evaluation were performed to evaluate the cartilage repair effects. Results In in vitro experiments, hUSCs had better capacity for proliferation, colony-forming, and migration compared to hBMSCs in the same passage, while hBMSCs had greater osteogenic, adipogenic, and chondrogenic abilities compared to hUSCs in the same passage. Both hUSCs and hBMSCs at passage 3 had the strongest potential for proliferation, colony-forming, and multilineage differentiation compared to cells in other passages. The ACM scaffolds loaded with hUSCs or hBMSCs both significantly promoted the repair of cartilage defects in the rabbit knee model at 12 weeks’ postimplantation, and the new tissue was mainly hyaline cartilage. However, there was no significant difference in cartilage repair effects between hUSCs and hBMSCs. Conclusions In in vitro experiments, hUSCs presented better capacity for proliferation, while hBMSCs had greater chondrogenic ability. However, hUSCs and hBMSCs had similar cartilage repair effects in vivo. Results indicated that hUSCs can be a stem cell alternative for cartilage regeneration and provide a powerful platform for cartilage tissue engineering and clinical transformation. Graphical abstract

Download Full-text

Effects of in vitro low oxygen tension preconditioning of buccal fat pad stem cells on in Vivo articular cartilage tissue repair

Life Sciences ◽

10.1016/j.lfs.2021.119728 ◽

2021 ◽

pp. 119728

Author(s):

Fatemeh Dehghani Nazhvani ◽

Leila Mohammadi Amirabad ◽

Arezo Azari ◽

Hamid Namazi ◽

Simzar Hosseinzadeh ◽

...

Keyword(s):

Stem Cells ◽

Articular Cartilage ◽

Tissue Repair ◽

Cartilage Tissue ◽

Low Oxygen ◽

Fat Pad ◽

Buccal Fat Pad ◽

Low Oxygen Tension

Download Full-text

Cell Source-Dependent In Vitro Chondrogenic Differentiation Potential of Mesenchymal Stem Cell Established from Bone Marrow and Synovial Fluid of Camelus dromedarius

Animals ◽

10.3390/ani11071918 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1918

Author(s):

Young-Bum Son ◽

Yeon Ik Jeong ◽

Yeon Woo Jeong ◽

Mohammad Shamim Hossein ◽

Per Olof Olsson ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Synovial Fluid ◽

Cartilage Regeneration ◽

Cartilage Tissue ◽

Chondrocyte Differentiation ◽

Differentiation Potential ◽

Proliferation Capacity

Mesenchymal stem cells (MSCs) are promising multipotent cells with applications for cartilage tissue regeneration in stem cell-based therapies. In cartilage regeneration, both bone marrow (BM-MSCs) and synovial fluid (SF-MSCs) are valuable sources. However, the cellular characteristics and chondrocyte differentiation potential were not reported in either of the camel stem cells. The in vitro chondrocyte differentiation competence of MSCs, from (BM and SF) sources of the same Camelus dromedaries (camel) donor, was determined. Both MSCs were evaluated on pluripotent markers and proliferation capacity. After passage three, both MSCs showed fibroblast-like morphology. The proliferation capacity was significantly increased in SF-MSCs compared to BM-MSCs. Furthermore, SF-MSCs showed an enhanced expression of transcription factors than BM-MSCs. SF-MSCs exhibited lower differentiation potential toward adipocytes than BM-MSCs. However, the osteoblast differentiation potential was similar in MSCs from both sources. Chondrogenic pellets obtained from SF-MSCs revealed higher levels of chondrocyte-specific markers than those from BM-MSCs. Additionally, glycosaminoglycan (GAG) content was elevated in SF-MSCs related to BM-MSCs. This is, to our knowledge, the first study to establish BM-MSCs and SF-MSCs from the same donor and to demonstrate in vitro differentiation potential into chondrocytes in camels.

Download Full-text