Spatiotemporal patterns of fibronectin distribution during embryonic development

Development ◽  
1981 ◽  
Vol 63 (1) ◽  
pp. 193-206
Author(s):  
Michael Melnick ◽  
Tina Jaskoll ◽  
Anna G. Brownell ◽  
Mary Macdougall ◽  
Conny Bessem ◽  
...  
Keyword(s):  
Limb Development ◽  
Spatial Organization ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Major Constituent ◽  
Alcian Blue Staining ◽  
In Ovo ◽  
Tissue Organization ◽  
Testicular Hyaluronidase

It has been suggested that an extracellular matrix - and cell surface - associated glycoprotein, fibronectin, plays a role in the positioning of cells in morphogenesis and in the maintenance of orderly tissue organization. In the present study the appearance and distribution of fibronectin during in ovo chick limb development has been investigated by indirect immunofluorescence techniques in H.H. stages 20–30. Fibronectin is not detectable until just prior to the transition from the morphogenetic to the cytodifferentiation phase of development. Beginning at H.H. stage 25, successive nonrandom patterns of fibronectin detection and distribution, which resemble the subsequent cartilaginous elements, precede overt chondrogenesis as detected by Alcian blue staining. This corresponds to the onset of the cytodifferentiation phase of limb development. As the accumulation of acidic proteoglycan increases in the cartilage matrix and the mesenchymal cells become more round in appearance, the presence of detectable fibronectin decreases and is ultimately seen only in the perichondria and basement membrane. However, predigestion of developed cartilage tissue with testicular hyaluronidase, prior to fibronectin staining, indicated that fibronectin remains a major constituent of cartilage matrix and is apparently masked by cartilagespecific proteoglycans. This study of chick limb development is consistent with the hypothesis that fibronectin may be a molecule that facilitates the spatial organization of cartilaginous primordia cytodifferentiation.

scholarly journals Silencing Smad7 Potentiates BMP2-induced Chondrogenic Differentiation and Inhibits Endochondral Ossification in Human Synovial Derived Mesenchymal Stem Cells

2020 ◽  
Author(s):  
pengcheng xiao ◽  
Zhenglin Zhu ◽  
Chengcheng Du ◽  
Yongsheng Zeng ◽  
junyi Liao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Cartilage Injuries

Abstract Background: Cartilage injuries pose formidable challenges for effective clinical management. Autologous stem cell-based therapies and transgene-enhanced cartilage tissue engineering may open new avenues for the treatment of cartilage injuries. Bone morphogenetic protein 2 (BMP2) is a promising chondrogenic growth factors for transgene-enhanced cartilage tissue engineering. However the BMP2 is failed to maintain a stable chondrogenic phenotype as it also induces robust endochondral ossification. Recently, human synovial derived mesenchymal stem cells (hSMSCs) arouse interested through the poor differentiation potential into osteogenic lineage. Smad7, a protein to antagonizes TGF-β/BMP signaling pathway has been discovered significant in the endochondral ossification. In the present study ,we further explore the effect of downregulate Smad7 in BMP2-induced chondrogenic differentiation of hSMSCs. Methods: hSMSCs were isolated from synovium of human knee joint through adhesion growth. In vitro and in vivo chondrogenic differentiation models of hSMSCs were constructed . Transgenes of BMP2, silencing Smad7 and Smad7 were expressed by adenoviral vectors. The osteogenic differentiation was detected by alkaline phosphatase staining, alizarin red staining. The chondrogenic differentiation was detected by alcian blue staining. Gene expression was determined by reverse transcription and quantitative real-time PCR (RT-qPCR), Immunofluorescence and immunohistochemistry. The subcutaneous stem cell implantation model was established and evaluated by micro-CT , h＆e staining, alcian blue staining and immunohistochemistry assay.Results: Compared to other MSCs, hSMSCs performed less of capability to osteogenic differentiation. But the occurrence of endochondral ossification is still inevasible during BMP2 induced cartilage formation. We found that silencing Smad7 enhanced the BMP2-induced chondrogenic differentiation of hSMSCs in vitro. Also, it leading to much less of hypertrophic differentiation. The subcutaneous stem cells implantation assays demonstrated silencing Smad7 potentiates BMP2-induced cartilage formation and inhibits endochondral ossification. Conclusion: This study strongly suggests that application of hSMSCs , cell scaffolds and silencing Smad7 can potentiate BMP2-induced chondrogenic differentiation and inhibit endochondral ossification. Thus, inhibit the expression of Smad7 in BMP2-induced hSMSCs differentiation may be a new strategy for cartilage tissue engineering.

scholarly journals The Effect of Human Platelet-Rich Plasma and L-Ascorbic Acid on Morphology, Proliferation, and Chondrogenesis Ability towards Human Adipose-Derived Stem Cells

2019 ◽  
Vol 3 (1) ◽  
pp. 26 ◽  
Author(s):  
Imam Rosadi ◽  
Karina Karina ◽  
Iis Rosliana ◽  
Siti Sobariah ◽  
Irsyah Afini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ascorbic Acid ◽  
Alcian Blue ◽  
Human Platelet ◽  
Platelet Rich Plasma ◽  
Cartilage Tissue ◽  
Population Doubling ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Adipose Derived Stem Cells

Background: Microtia is a congenital malformation in the external ear due to cartilage defect. Adipose-derived stem cells (ADSC) is promising cells to develop cartilage tissue engineering for microtia. In this study, we focused on proliferation and chondrogenesis of ADSC in three different media, which consist of 10% fetal bovine serum (FBS), 10% FBS with L-ascorbic acid, and 10% human platelet rich plasma (PRP). Methods: ADSC were induced to differentiate into adipocytes, chondrocyte and osteocytes. ADSC morphology, proliferation and population doubling time was compared in three different media and analysed. Observation and alcian blue staining were done every 7 days to assess chondrogenic potency of ADSC from each treatment.Results: Isolated ADSC were able to differentiate into adipocytes, osteocytes and chondrocytes. ADSC in all group have fibroblast-like morphology, but cells in 10% FBS and 10% FBS with LAA group were flattened and larger. ADSC in 10% PRP group proliferates faster than 10% FBS with and without LAA. PDT values of ADSC were 34 hours, 44 hours and 48 hours, respectively for 10% PRP, 10% FBS with LAA and 10% FBS group. Alcian blue staining revealed that ADSC in 10% FBS with LAA and 10% PRP were able to proceed to chondrogenesis when cultured time were prolong up to 21 days, but not with ADSC in 10% FBS. Conclusion: We conclude that adding 10% FBS with LAA or 10% PRP into medium culture can support proliferation and chondrogenesis of ADSC. Key words: human ADSC, PRP, L-ascorbic acid, proliferation, chondrogenesis

scholarly journals Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides

1984 ◽  
Vol 221 (3) ◽  
pp. 707-716 ◽  
Author(s):  
M K Cowman ◽  
M F Slahetka ◽  
D M Hittner ◽  
J Kim ◽  
M Forino ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Unit Structure ◽  
Electrophoretic Patterns ◽  
Sulphated Glycosaminoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Testicular Hyaluronidase

Oligosaccharide fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. An extensive ladder-like set of bands is observed for partial testicular hyaluronidase digests of chondroitin 4- or 6-sulphate, and for dermatan sulphate. Co-electrophoresis of purified oligosaccharides has established that the major bands of these patterns represent fragments differing in chain length by one disaccharide unit, with the smallest fragments having the greatest mobility. Additional minor bands, representing heterogeneity in the repeating unit structure, are also observed. There are slight differences in the mobilities of oligosaccharides derived from the three major types of sulphated glycosaminoglycans. Alcian Blue is employed for visualization of the digest fragments. Sample loads of 5-10 micrograms per band appear optimum. The smallest oligosaccharide which may be stained by this method is the hexasaccharide. After consideration of this effect, a good correlation is found to exist between densitometric scans of the gel-electrophoretic patterns and gel-filtration chromatographic profiles based on uronic acid concentration.

scholarly journals Hybrid Hydrogel Composed of Hyaluronic Acid, Gelatin, and Extracellular Cartilage Matrix for Perforated TM Repair

2021 ◽  
Vol 9 ◽  
Author(s):  
Yili Wang ◽  
Feng Wen ◽  
Xueting Yao ◽  
Lulu Zeng ◽  
Jiaming Wu ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Cartilage Matrix ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Hybrid Hydrogel ◽  
Composite Hydrogels ◽  
Further Development ◽  
Perforated Tympanic Membrane

A novel series of composite hydrogels, built from the three components 1), hyaluronic acid methacryloyl (HAMA); 2), gelatin methacryloyl (GelMA), and 3), extracellular cartilage matrix (ECM), was prepared and studied regarding the possible utility in the surgical repair of damaged (perforated) tympanic membrane (TM). Noteworthy is component 3), which was harvested from the ribs of α-1,3-galactosidyltransferase-knockout (α-1,3 GalT-KO) pigs. The absence of α-1,3-galactosyl glycoprotein is hypothesized to prevent rejection due to foreign-body immunogenicity. The composite hydrogels were characterized by various aspects, using a variety of physicochemical techniques: aqueous swelling, structural degradation, behavior under compression, and morphology, e.g., in vitro biocompatibility was assessed by the CCK-8 and live–dead assays and through cytoskeleton staining/microscopy. Alcian blue staining and real-time PCR (RT-PCR) were performed to examine the chondrogenic induction potential of the hydrogels. Moreover, a rat TM defect model was used to evaluate the in vivo performance of the hydrogels in this particular application. Taken together, the results from this study are surprising and promising. Much further development work will be required to make the material ready for surgical use.

scholarly journals Chondrogenic Differentiation of Mesenchymal Stem Cells in Three-Dimensional Chitosan Film Culture

2017 ◽  
Vol 26 (3) ◽  
pp. 417-427 ◽  
Author(s):  
Tsai-Jung Lu ◽  
Fang-Yao Chiu ◽  
Hsiao-Ying Chiu ◽  
Ming-Chau Chang ◽  
Shih-Chieh Hung
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Chitosan Film ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Cartilage Defects ◽  
Induction Medium ◽  
Alcian Blue Staining ◽  
Film Culture

Articular cartilage has a very limited capacity for self-repair, and mesenchymal stem cells (MSCs) have the potential to treat cartilage defects and osteoarthritis. However, in-depth mechanistic studies regarding their applications are required. Here we demonstrated the use of chitosan film culture for promoting chondrogenic differentiation of MSCs. We found that MSCs formed spheres 2 days after seeding on dishes coated with chitosan. When MSCs were induced in a chondrogenic induction medium on chitosan films, the size of the spheres continuously increased for up to 21 days. Alcian blue staining and immunohistochemistry demonstrated the expression of chondrogenic proteins, including aggrecan, type II collagen, and type X collagen at 14 and 21 days of differentiation. Importantly, chitosan, with a medium molecular weight (size: 190–310 kDa), was more suitable than other sizes for inducing chondrogenic differentiation of MSCs in terms of sphere size and expression of chondrogenic proteins and endochondral markers. We identified that the mechanistic target of rapamycin (mTOR) signaling and its downstream S6 kinase (S6K)/S6 were activated in chitosan film culture compared to that of monolayer culture. The activation of mTOR/S6K was continuously upregulated from days 2 to 7 of differentiation. Furthermore, we found that mTOR/S6K signaling was required for chondrogenic differentiation of MSCs in chitosan film culture through rapamycin treatment and mTOR knockdown. In conclusion, we showed the suitability of chitosan film culture for promoting chondrogenic differentiation of MSCs and its potential in the development of new strategies in cartilage tissue engineering.

scholarly journals Knockout of miR-21-5p alleviates cartilage matrix degradation by targeting Gdf5 in temporomandibular joint osteoarthritis

2020 ◽  
Vol 9 (10) ◽  
pp. 689-700
Author(s):  
Aobo Zhang ◽  
Shixing Ma ◽  
Lingyu Yuan ◽  
Shichao Wu ◽  
Shaopeng Liu ◽  
...  
Keyword(s):  
Temporomandibular Joint ◽  
Cartilage Matrix ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Histological Structure ◽  
Matrix Degradation ◽  
Matrix Metalloproteinase 13 ◽  
Mouse Tissues ◽  
Temporomandibular Joint Osteoarthritis ◽  
Safranin O

Aims The study aimed to determine whether the microRNA miR21-5p (MiR21) mediates temporomandibular joint osteoarthritis (TMJ-OA) by targeting growth differentiation factor 5 (Gdf5). Methods TMJ-OA was induced in MiR21 knockout (KO) mice and wild-type (WT) mice by a unilateral anterior crossbite (UAC) procedure. Mouse tissues exhibited histopathological changes, as assessed by: Safranin O, toluidine blue, and immunohistochemistry staining; western blotting (WB); and quantitative real-time polymerase chain reaction (RT-qPCR). Mouse condylar chondrocytes were transfected with a series of MiR21 mimic, MiR21 inhibitor, Gdf5 siRNA (si-GDF5), and flag-GDF5 constructs. The effects of MiR-21 and Gdf5 on the expression of OA related molecules were evaluated by immunofluorescence, alcian blue staining, WB, and RT-qPCR. Results UAC altered the histological structure and extracellular matrix content of cartilage in the temporomandibular joint (TMJ), and KO of MiR21 alleviated this effect (p < 0.05). Upregulation of MiR21 influenced the expression of TMJ-OA related molecules in mandibular condylar chondrocytes via targeting Gdf5 (p < 0.05). Gdf5 overexpression significantly decreased matrix metalloproteinase 13 (MMP13) expression (p < 0.05) and reversed the effects of MiR21 (p < 0.05). Conclusion MiR21, which acts as a critical regulator of Gdf5 in chondrocytes, regulates TMJ-OA related molecules and is involved in cartilage matrix degradation, contributing to the progression of TMJ-OA. Cite this article: Bone Joint Res 2020;9(10):689–700.

scholarly journals COMPARISON OF THE EFFECTS OF PAPAIN AND VITAMIN A ON CARTILAGE

1960 ◽  
Vol 111 (5) ◽  
pp. 705-718 ◽  
Author(s):  
Lewis Thomas ◽  
Robert T. McCluskey ◽  
Jacobus L. Potter ◽  
Gerald Weissmann
Keyword(s):  
Vitamin A ◽  
Chondroitin Sulfate ◽  
Cartilage Matrix ◽  
Epiphyseal Cartilage ◽  
Blue Staining ◽  
Daily Injection ◽  
Alcian Blue Staining ◽  
The Matrix ◽  
Cartilage Cells ◽  
Ear Cartilage

The administration of large amounts of vitamin A to rabbits has been shown to result in depletion of cartilage matrix. The normal basophilic, metachromatic, and Alcian blue staining properties of the matrix are lost, especially in articular and epiphyseal cartilage. The cartilage cells remain intact, but are reduced in size. These changes sometimes appeared as early as 48 hours after the initiation of daily injection of 1 million units of vitamin A, and were usually well established by 5 days. Some rabbits failed to show changes in cartilage, even after 5 daily injections. Increased amounts of material presumed to be chondroitin sulfate were present in the sera of vitamin A-treated rabbits, usually by 72 hours after the first injection. This was demonstrated by a turbidimetric procedure using hexamminecobaltic chloride. In rabbits given sulfur-35 (Na2S35O4) 5 days before the initiation of vitamin A treatment, it was shown that sulfur-35 was lost from articular and epiphyseal cartilage. This was associated with an increase in the non-dialyzable sulfur-35 in both serum and in the cobalt-precipitable material. These rabbits also excreted more sulfur-35 than rabbits not given vitamin A. There was a reduction in sulfur-35 activity in chondromucoprotein extracted from the ear cartilage of vitamin A-treated rabbits. The changes are interpreted as indicating that the administration of large amounts of vitamin A to rabbits results in removal of chondroitin sulfate from cartilage matrix. The administration of small amounts of crude papain causes histologic changes in cartilage that are remarkably similar to those seen in vitamin A-treated rabbits. The possibility is suggested that the changes in cartilage produced by administration of vitamin A to rabbits may be the result of activation of a proteolytic enzyme or enzymes, with properties similar to those of papain.

scholarly journals Transient supplementation of growth factor TGF-β1 effectively initiates chondrogenic redifferentiation of human chondrocytes

2017 ◽  
Vol 3 (2) ◽  
pp. 383-387 ◽  
Author(s):  
Bettina Hiemer ◽  
Kathleen Achenbach ◽  
Juliane Pasold ◽  
Andreas Wree ◽  
Rainer Bader
Keyword(s):  
Cell Proliferation ◽  
Human Chondrocytes ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Proteoglycan Synthesis ◽  
Alcian Blue Staining ◽  
Cell Clusters ◽  
Cell Clustering ◽  
Tgf Β1

AbstractCartilage tissue is avascular with less regeneration potential and therefore, cartilage regeneration is still a major challenge for therapeutic approaches. Commonly used treatment strategies involve the transplantation of autologous chondrocytes into the defect. Before that, it is required to increase the cell number in vitro resulting in unwanted chondrocyte dedifferentiation. This could impair subsequent tissue regeneration. Both growth factors TGF-ß1 and IGF-1 are used as strong inducer of chondrogenic redifferentiation, however, a controlled application of TGF-ß1 is essential to avoid adverse effects. Therefore, in the present study, we investigated the time-dependent influence of TGF-ß1 administration on chondrocyte redifferentiation.Human chondrocytes were embedded in alginate and cultured in serum-free DMEM containing ascorbic acid, dexamethasone, ITSTM and IGF-1. TGF-β1 was supplemented for 3, 7 and 21 days. Afterwards, cell viability and synthesis of extracellular matrix (ECM) proteins was analyzed by histological staining.Live/dead staining of chondrocytes incubated with TGF-β1 for 21 days displayed an enhanced proliferation and formation of cell clusters resulting in excessive outgrowth of fibroblastic-like cells. However, exposure to TGF-β1 over only 7 days caused also cell clustering with moderate cell proliferation. Additionally, after 21 days of cultivation proteoglycan synthesis was identified by alcian blue staining after both TGF-β1 supplementation for 21 and also 7 days. Aggrecan was also detected in the periphery of the cell clusters after TGF-β1 incubation for only 7 days. Chondrocytes lacked proteoglycan expression after three-day TGF-β1 administration.We could show, that prolonged administration of TGF-β1 results in massive proliferation of chondrocytes which is accompanied by cell outgrowth. We found that TGF-ß1 exposure for seven days is sufficient for achievement of cell clustering without excessive cell proliferation, which is important for inducing subsequent chondrogenic differentiation. Results indicate that even an initial TGF-β1 administration could be sufficient for inducing chondrocyte proliferation and differentiation in vitro.

scholarly journals miR-223 promotes cartilage differentiation of bone marrow-derived mesenchymal stem cells and protects against osteoarthritis by suppressing NLRP-3 expression

2020 ◽  
Author(s):  
Nan Min ◽  
Jie Ma ◽  
Lei Shi ◽  
Lin Wang ◽  
Chi Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Group Treatment ◽  
Chondrogenic Differentiation ◽  
Cruciate Ligament ◽  
Treatment Protocol ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Alcian Blue Staining

IntroductionThe present investigation evaluates the role of miR-223 mimic in the treatment of osteoarthritis (OA) and postulates the possible molecular mechanism of its action.Material and methodsBone marrow-derived mesenchymal stem cells (BMSCs) were isolated from rats and cultured in chondrogenic medium to stimulate the differentiation of chondrocytes. Alcian blue staining was performed to determine the chondrogenic differentiation and expression of miR-223 in the BMSCs. Moreover, expression of NLR family pyrin domain containing 3 (NLRP-3), matrix metallopeptidase-13 (MMP-13) and collagen (Col II) was determined in miR-223 mimic and inhibitor treated BMSCs. OA was induced by injecting anterior cruciate ligament transection in rats followed by further treatment with the miR-223 mimic for the period of the treatment protocol. Level and expression of inflammatory cytokines were estimated in the cartilage tissue of OA rats. Moreover, immunohistochemical analysis and histopathology study were also performed.ResultsData of the study reveal that expression of miR-223 was higher in chondrogenic differentiated BMSCs than normal. Expression of MMP-13 and NLRP-3 was lower, and expression of Col II was higher in miR-223 mimic treated BMSCs than normal. Moreover, data of the in-vivo study indicate that the expression level of cytokines was lower in the cartilage tissue of the miR-223 mimic treated group than the OA group. Treatment with the miR-223 mimic ameliorates the altered histopathology and expression of NLRP-3 in the cartilage tissue of OA rats.ConclusionsData of the study reveal that the miR-223 mimic enhances the chondrogenic differentiation of BMSCs by regulating the NLRP-3/IL-18/TGF-b pathway.

scholarly journals Silencing Smad7 potentiates BMP2-induced chondrogenic differentiation and inhibits endochondral ossification in human synovial-derived mesenchymal stromal cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Pengcheng Xiao ◽  
Zhenglin Zhu ◽  
Chengcheng Du ◽  
Yongsheng Zeng ◽  
Junyi Liao ◽  
...  
Keyword(s):  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Pcr Analysis ◽  
Alcian Blue Staining ◽  
Rt Pcr ◽  
Cell Implantation

Abstract Background Bone morphogenetic protein 2 (BMP2) is a promising chondrogenic growth factor for cartilage tissue-engineering, but it also induces robust endochondral ossification. Human synovial-derived mesenchymal stromal cells (hSMSCs) have attracted great interest due to their poor potential for differentiation into osteogenic lineages. Smad7 plays a significant in the endochondral ossification. In this study, we explored a new method to amplify the BMP2-induced chondrogenic differentiation of hSMSCs by downregulating Smad7 and applying a cellular scaffold. Methods hSMSCs were isolated from human knee joint synovium from 3 donors through adhesion growth. In vitro and in vivo models of the chondrogenic differentiation of hSMSCs were established. Transgenic expression of BMP2 and silencing of Smad7 and Smad7 was achieved by adenoviral vectors. The osteogenic differentiation was detected by alkaline phosphatase staining, alizarin red staining, and RT-PCR analysis of the osteogenic genes RUNX2, Osterix, and Osteocalcin. The chondrogenic differentiation was detected by Alcian blue staining and RT-PCR analysis of the chondrogenic genes SOX9, COL2, and aggrecan. Hypertrophic differentiation was detected by the markers COL10 and MMP13. A subcutaneous stem cell implantation model was established with polyethylene glycol citrate-co-N-isopropylacrylamide (PPCN) scaffolds and athymic nude mice (3/group, 4–6 week-old female) and evaluated by micro-CT, H&E staining, and Alcian blue staining. An immunohistochemistry assay was used to detected COL1 and COL2, and an immunofluorescence assay was used to detect COL10 and MMP13. Results These hSMSCs identified by flow cytometry. These hSMSCs exhibited lower osteo-differentiation potential than iMads and C3H10T1/2-cells. When Smad7 was silenced in BMP2-induced hSMSCs, the chondrogenic differentiation genes SOX9, COL2, and aggrecan were enhanced in vitro. Additionally, it silencing Smad7 led to a decrease in the hypertrophic differentiation genes COL10 and MMP13. In subcutaneous stem cell implantation assays, immunofluorescence and immunohistochemical staining demonstrated that silencing Smad7 increased the number of COL2-positive cells and decreased the expression of COL1, COL10, and MMP13. Conclusion This study suggests that the application of hSMSCs, cell scaffolds, and silencing Smad7 can potentiate BMP2-induced chondrogenic differentiation and inhibit endochondral ossification. Thus, inhibiting the expression of Smad7 in BMP2-induced hSMSC differentiation may be a new strategy for cartilage tissue-engineering.

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