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1.
The feasibility of using genipin cross-linked type II collagen scaffold with rabbit bone marrow mesenchymal stem cells (RBMSCs)
to repair cartilage defect was herein studied. Induction of RBMSCs into chondrocytic phenotype on type II collagen scaffold
in vitro was conducted using TGF-β 3 containing medium. After 3-weeks of induction, chondrocytic behavior, including marker
genes expression and specific extracellular matrix (ECM) secretion, was observed. In the in vivo evaluation experiment, the
scaffolds containing RBMSCs without prior induction were autologous implanted into the articular cartilage defects made by
subchondral drilling. The repairing ability was evaluated. After 2 months, chondrocyte-like cells with lacuna structure and
corresponding ECM were found in the repaired sites without apparent inflammation. After 24 weeks, we could easily find cartilage
structure the same with normal cartilage in the repair site. In conclusion, it was shown that the scaffolds in combination
of in vivo conditions can induce RBMSCs into chondrocytes in repaired area and would be a possible method for articular cartilage
repair in clinic and cartilage tissue engineering. 相似文献
2.
Filová E Rampichová M Handl M Lytvynets A Halouzka R Usvald D Hlucilová J Procházka R Dezortová M Rolencová E Kostáková E Trc T Stastný E Kolácná L Hájek M Motlík J Amler E 《Physiological research / Academia Scientiarum Bohemoslovaca》2007,56(Z1):S5-S16
The potential of novel scaffold containing sodium hyaluronate, type I collagen, and fibrin was investigated in the regeneration of osteochondral defects in miniature pigs. Both autologous chondrocyte-seeded scaffolds and non-seeded scaffolds were implanted into two defects located in the non-weight-bearing zone of the femoral trochlea (defect A was located more distally and medially, defect B was located more proximally and laterally). Control defects were left untreated. Twelve weeks after the operation, the knees were evaluated in vivo using MRI. Six months after the implantation, the defects were analyzed using MRI, histological, and immunohistochemical analysis. In the A defects of chondrocyte-seeded scaffold group, hyaline cartilage and fibrocartilage was formed, containing type II collagen, acidic and neutral glycosaminoglycans while the non-seeded scaffold group was predominantly filled with fibrocartilage. Defects in the control group were predominantly filled with fibrous tissue. Histomorphometric analysis of photomicrographs revealed a significantly higher amount of hyaline cartilage in the cell-seeded scaffold group in A defects than in other groups. Both scaffold groups in A defects showed significantly less fibrous tissue than cell-seeded defects B and the control group. Both histological and MRI analysis proved that the novel composite scaffold has a potential to regenerate osteochondral defects within six months. 相似文献
3.
Tissue engineering (TE) has been proven usefulness in cartilage defect repair. For effective cartilage repair, the structural orientation of the cartilage scaffold should mimic that of native articular cartilage, as this orientation is closely linked to cartilage mechanical functions. Using thermal-induced phase separation (TIPS) technology, we have fabricated an oriented cartilage extracellular matrix (ECM)-derived scaffold with a Young''s modulus value 3 times higher than that of a random scaffold. In this study, we test the effectiveness of bone mesenchymal stem cell (BMSC)-scaffold constructs (cell-oriented and random) in repairing full-thickness articular cartilage defects in rabbits. While histological and immunohistochemical analyses revealed efficient cartilage regeneration and cartilaginous matrix secretion at 6 and 12 weeks after transplantation in both groups, the biochemical properties (levels of DNA, GAG, and collagen) and biomechanical values in the oriented scaffold group were higher than that in random group at early time points after implantation. While these differences were not evident at 24 weeks, the biochemical and biomechanical properties of the regenerated cartilage in the oriented scaffold-BMSC construct group were similar to that of native cartilage. These results demonstrate that an oriented scaffold, in combination with differentiated BMSCs can successfully repair full-thickness articular cartilage defects in rabbits, and produce cartilage enhanced biomechanical properties. 相似文献
4.
Lohan A Marzahn U El Sayed K Haisch A Kohl B Müller RD Ertel W Schulze-Tanzil G John T 《Histochemistry and cell biology》2011,136(1):57-69
Implantation of tissue-engineered heterotopic cartilage into joint cartilage defects might be an alternative approach to improve
articular cartilage repair. Hence, the aim of this study was to characterize and compare the quality of tissue-engineered
cartilage produced with heterotopic (auricular, nasoseptal and articular) chondrocytes seeded on polyglycolic acid (PGA) scaffolds
in vitro and in vivo using the nude mice xenograft model. PGA scaffolds were seeded with porcine articular, auricular and
nasoseptal chondrocytes using a dynamic culturing procedure. Constructs were pre-cultured 3 weeks in vitro before being implanted
subcutaneously in nude mice for 1, 6 or 12 weeks, non-seeded scaffolds were implanted as controls. Heterotopic neo-cartilage
quality was assessed using vitality assays, macroscopical and histological scoring systems. Neo-cartilage formation could
be observed in vitro in all PGA associated heterotopic chondrocytes cultures and extracellular cartilage matrix (ECM) deposition
increased in vivo. The 6 weeks in vivo incubation time point leads to more consistent results for all cartilage species, since
at 12 weeks in vivo construct size reductions were higher compared with 6 weeks except for auricular chondrocytes PGA cultures.
Some regressive histological changes could be observed in all constructs seeded with all chondrocytes subspecies such as cell-free
ECM areas. Particularly, but not exclusively in nasoseptal chondrocytes PGA cultures, ossificated ECM areas appeared. Elastic
fibers could not be detected within any neo-cartilage. The neo-cartilage quality did not significantly differ between articular
and non-articular chondrocytes constructs. Whether tissue-engineered heterotopic neo-cartilage undergoes sufficient transformation,
when implanted into joint cartilage defects requires further investigation. 相似文献
5.
目的:探讨采用软骨细胞外基质材料制备的定向结构软骨支架复合软骨细胞,在体外静态培养条件下生成组织工程软骨的可能性。方法:制备牛关节软骨细胞外基质材料,利用温度梯度热诱导相分离技术构建具备垂直定向孔道结构的软骨支架,同时采用传统冷冻干燥方法制备非定向支架,检测两组支架的力学性能;提取兔关节软骨细胞,分别接种两组支架,体外静态培养2周及4周后取材,对构建的组织工程软骨进行组织切片染色、生物化学分析及生物力学检测。结果:定向软骨支架的压缩弹性模量数值明显高于非定向软骨支架,体外培养时定向支架上种子细胞在3-9d内增殖高于非定向支架,差异有统计学意义(P〈0.05);体外静态培养4周后形成的两组新生组织工程软骨进行软骨特异性染色均呈阳性,在定向组新生软骨切片中在垂直方向上可见大量呈规则平行排列的粗大胶原纤维,两组新生软骨的生物化学检测包括总DNA、总GAG及总胶原含量差异无统计学意义(P〉0.05)。定向组织工程软骨压缩弹性模量在2周及4周时均高于非定向组织工程软骨,差异有统计学意义(P〈0.05)。但两组组织工程软骨上述指标均显著低于正常关节软骨(P〈0.05)。结论:软骨细胞外基质材料制备的定向结构软骨支架复合软骨细胞,在体外静态培养条件下能够成功生成具有定向纤维结构的组织工程软骨,并可以有效促进新生软骨组织力学性能的提升,在软骨组织工程中具有良好的应用前景。 相似文献
6.
Silvia Díaz-Prado Mª Esther Rendal-Vázquez Emma Muiños-López Tamara Hermida-Gómez Margarita Rodríguez-Cabarcos Isaac Fuentes-Boquete Francisco J. de Toro Francisco J. Blanco 《Cell and tissue banking》2010,11(2):183-195
The human amniotic membrane (HAM) is an abundant and readily obtained tissue that may be an important source of scaffold for
transplanted chondrocytes in cartilage regeneration in vivo. To evaluate the potential use of cryopreserved HAMs as a support
system for human chondrocytes in human articular cartilage repair. Chondrocytes were isolated from human articular cartilage,
cultured and grown on the chorionic basement membrane side of HAMs. HAMs with chondrocytes were then used in 44 in vitro human
osteoarthritis cartilage repair trials. Repair was evaluated at 4, 8 and 16 weeks by histological analysis. Chondrocytes cultured
on the HAM revealed that cells grew on the chorionic basement membrane layer, but not on the epithelial side. Chondrocytes
grown on the chorionic side of the HAM express type II collagen but not type I, indicating that after being in culture for
3–4 weeks they had not de-differentiated into fibroblasts. In vitro repair experiments showed formation on OA cartilage of
new tissue expressing type II collagen. Integration of the new tissue with OA cartilage was excellent. The results indicate
that cryopreserved HAMs can be used to support chondrocyte proliferation for transplantation therapy to repair OA cartilage. 相似文献
7.
Ossendorf C Kaps C Kreuz PC Burmester GR Sittinger M Erggelet C 《Arthritis research & therapy》2007,9(2):R41
Autologous chondrocyte implantation (ACI) is an effective clinical procedure for the regeneration of articular cartilage defects. BioSeed-C is a second-generation ACI tissue engineering cartilage graft that is based on autologous chondrocytes embedded in a three-dimensional bioresorbable two-component gel-polymer scaffold. In the present prospective study, we evaluated the short-term to mid-term efficacy of BioSeed-C for the arthrotomic and arthroscopic treatment of posttraumatic and degenerative cartilage defects in a group of patients suffering from chronic posttraumatic and/or degenerative cartilage lesions of the knee. Clinical outcome was assessed in 40 patients with a 2-year clinical follow-up before implantation and at 3, 6, 12, and 24 months after implantation by using the modified Cincinnati Knee Rating System, the Lysholm score, the Knee injury and Osteoarthritis Outcome Score, and the current health assessment form (SF-36) of the International Knee Documentation Committee, as well as histological analysis of second-look biopsies. Significant improvement (p < 0.05) in the evaluated scores was observed at 1 and/or 2 years after implantation of BioSeed-C, and histological staining of the biopsies showed good integration of the graft and formation of a cartilaginous repair tissue. The Knee injury and Osteoarthritis Outcome Score showed significant improvement in the subclasses pain, other symptoms, and knee-related quality of life 2 years after implantation of BioSeed-C in focal osteoarthritic defects. The results suggest that implanting BioSeed-C is an effective treatment option for the regeneration of posttraumatic and/or osteoarthritic defects of the knee. 相似文献
8.
In this study, we designed a chitosan/alginate/hydroxyapatite scaffold as a carrier for recombinant BMP-2 (CAH/B2), and evaluated the release kinetics of BMP-2. We evaluated the effect of the CAH/B2 scaffold on the viability and differentiation of bone marrow mesenchymal stem cells (MSCs) by scanning electron microscopy, MTS, ALP assay, alizarin-red staining and qRT-PCR. Moreover, MSCs were seeded on scaffolds and used in a 8 mm rat calvarial defect model. New bone formation was assessed by radiology, hematoxylin and eosin staining 12 weeks postoperatively. We found the release kinetics of BMP-2 from the CAH/B2 scaffold were delayed compared with those from collagen gel, which is widely used for BMP-2 delivery. The BMP-2 released from the scaffold increased MSC differentiation and did not show any cytotoxicity. MSCs exhibited greater ALP activity as well as stronger calcium mineral deposition, and the bone-related markers Col1α, osteopontin, and osteocalcin were upregulated. Analysis of in vivo bone formation showed that the CAH/B2 scaffold induced more bone formation than other groups. This study demonstrates that CAH/B2 scaffolds might be useful for delivering osteogenic BMP-2 protein and present a promising bone regeneration strategy. 相似文献
9.
A cell leakproof porous poly(DL ‐lactic‐co‐glycolic acid) (PLGA)‐collagen hybrid scaffold was prepared by wrapping the surfaces of a collagen sponge except the top surface for cell seeding with a bi‐layered PLGA mesh. The PLGA‐collagen hybrid scaffold had a structure consisting of a central collagen sponge formed inside a bi‐layered PLGA mesh cup. The hybrid scaffold showed high mechanical strength. The cell seeding efficiency was 90.0% when human mesenchymal stem cells (MSCs) were seeded in the hybrid scaffold. The central collagen sponge provided enough space for cell loading and supported cell adhesion, while the bi‐layered PLGA mesh cup protected against cell leakage and provided high mechanical strength for the collagen sponge to maintain its shape during cell culture. The MSCs in the hybrid scaffolds showed round cell morphology after 4 weeks culture in chondrogenic induction medium. Immunostaining demonstrated that type II collagen and cartilaginous proteoglycan were detected in the extracellular matrices. Gene expression analyses by real‐time PCR showed that the genes encoding type II collagen, aggrecan, and SOX9 were upregulated. These results indicated that the MSCs differentiated and formed cartilage‐like tissue when being cultured in the cell leakproof PLGA‐collagen hybrid scaffold. The cell leakproof PLGA‐collagen hybrid scaffolds should be useful for applications in cartilage tissue engineering. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 相似文献
10.
Sivasami Pulavendran Gurunathen Thiyagarajan 《Biotechnology and Bioprocess Engineering》2011,16(2):393-399
Stem cell-based tissue engineering holds much hope for the development of multifunctional tissues to replace diseased organs.
The attachment and survival of stem cells on a three-dimensional (3D) scaffold must be enhanced for faster progression of
stem cell based tissue engineering. This study evaluate the stability of mesenchymal stem cells (MSCs) in 3D porous scaffolds
composed of a collagen and chitosan blend impregnated with epidermal growth factor incorporated chitosan nanoparticles (EGF-CNP).
The EGF-CNP scaffolds were characterized by transmission electron microscopy, which revealed that the nanoparticles were round
in shape and 20 ∼ 50 nm in size. The scaffolds were prepared by freeze drying. A Fourier-transform infrared spectrum study
revealed that the linkage between collagen and chitosan was through an ionic interaction. Thermal analysis and degradation
studies showed that the scaffold could be used in tissue engineering application. MSCs proliferated well in the EGF-CNP impregnated
scaffold. A scanning electron microscope study showed anchored and elongated MSCs on the EGF-CNP impregnated scaffold. A 3D
biodegradable collagen chitosan scaffold impregnated with EGF-CNP is a promising transportable candidate for MSC-based tissue
engineering, and this scaffold could be used as an in vitro model for subsequent clinical applications. 相似文献
11.
Tissue transglutaminase_variant 2-transduced mesenchymal stem cells and their chondrogenic potential
Ayse Ceren Calikoglu Koyuncu Ayse Hande Nayman Dilek Telci Gamze Torun Kose 《Biotechnology and bioengineering》2020,117(6):1839-1852
As cartilage is incapable of self-healing upon severe degeneration because of the lack of blood vessels, cartilage tissue engineering is gaining importance in the treatment of cartilage defects. This study was designed to improve cartilage tissue regeneration by expressing tissue transglutaminase variant 2 (TGM2_v2) in mesenchymal stem cells (MSC) derived from bone marrow of rats. For this purpose, rat MSCs transduced with TGM2_v2 were grown and differentiated on three-dimensional polybutylene succinate (PBSu) and poly-l -lactide (PLLA) blend scaffolds. The transduced cells could not only successfully express the short form transglutaminase-2, but also deposited the protein onto the scaffolds. In addition, they could spontaneously produce cartilage-specific proteins without any chondrogenic induction, suggesting that TGM2_v2 expression provided the cells the ability of chondrogenic differentiation. PBSu:PLLA scaffolds loaded with TGM2_v2 expressing MSCs could be used in repair of articular cartilage defects. 相似文献
12.
In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel 总被引:2,自引:0,他引:2
Yokoyama A Sekiya I Miyazaki K Ichinose S Hata Y Muneta T 《Cell and tissue research》2005,322(2):289-298
Graft implantation is one of the more popular procedures for repairing cartilage defects; however, sacrifices of the donor
site have been an issue. Mesenchymal stem cells (MSCs) are a fascinating source for regenerative medicine because they can
be harvested in a less invasive manner and are easily isolated and expanded, with multipotentiality including chondrogenesis.
MSCs can be isolated from various adult mesenchymal tissues including synovium. Here, we attempted to form cartilage from
the composites of synovium-derived MSCs with collagen gel in vitro. After 21 days of culture, the composites had increased
their cartilage matrix, as demonstrated by toluidine blue staining and immunohistochemistry for type II collagen. The composites
consisting of 5×107 and 108 cells/ml in gel were richer in proteoglycans than those consisting of lower cell densities. After 1 day, MSCs/gel composites
contracted and the diameter decreased by 30%; however, they were stable thereafter. Round cells with short processes producing
collagen fibrils showing a similar morphology to that of chondrocytes were seen in the composites by transmission electron
microscopy. During composite culture, chondroitin sulfate and mRNA expression for cartilage-related genes increased, demonstrating
cartilage maturation. Using an optimized method, we obtained cartilage discs with a diameter of 7 mm and a thickness of 500 μm.
Our procedure should thus make it possible to produce a large cartilage matrix in vitro. The tissue engineering of autologous
cartilage from the composites of synovium-derived MSCs with collagen gel in vitro for transplantation may be a future alternative
to graft implantation for patients with cartilage defects.
This study is supported in part by grants from the Japanese Society for the Promotion of Science (16591478), the Japanese
Orthopaedics and Traumatology Foundation, and the Nakatomi Foundation to I.S., and the Japanese Society for the Promotion
of Science (16591477), the Japanese Sports Medicine Foundation, the Japanese Latest Osteoarthritis Society, and the Center
of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone in Tokyo Medical
and Dental University to T.M. 相似文献
13.
Objective
Massive bone allografts are frequently used in orthopedic reconstructive surgery, but carry a high failure rate of approximately 25%. We tested whether treatment of graft with mesenchymal stem cells (MSCs) can increase the integration of massive allografts (hemi-mandible) in a large animal model.Methods
Thirty beagle dogs received surgical left-sided hemi-mandibular defects, and then divided into two equal groups. Bony defects of the control group were reconstructed using allografts only. Those of the experimental group were reconstructed using allogenic mandibular scaffold-loaded autologous MSCs. Beagles from each group were killed at4 (n = 4), 12 (n = 4), 24 (n = 4) or 48 weeks (n = 3) postoperatively. CT and micro-CT scans, histological analyses and the bone mineral density (BMD) of transplants were used to evaluate defect reconstruction outcomes.Results
Gross and CT examinations showed that the autologous bone grafts had healed in both groups. At 48 weeks, the allogenic mandibular scaffolds of the experimental group had been completely replaced by new bone, which has a smaller surface area to that of the original allogenic scaffold, whereas the scaffold in control dogs remained the same size as the original allogenic scaffold throughout. At 12 weeks, the BMD of the experimental group was significantly higher than the control group (p<0.05), and all micro-architectural parameters were significantly different between groups (p<0.05). Histological analyses showed almost all transplanted allogeneic bone was replaced by new bone, principally fibrous ossification, in the experimental group, which differed from the control group where little new bone formed.Conclusions
Our study demonstrated the feasibility of MSC-loaded allogenic mandibular scaffolds for the reconstruction of hemi-mandibular defects. Further studies are needed to test whether these results can be surpassed by the use of allogenic mandibular scaffolds loaded with a combination of MSCs and osteoinductive growth factors. 相似文献14.
Tissue engineering of human cartilage in bioreactors using single and composite cell-seeded scaffolds 总被引:5,自引:0,他引:5
Chondrocytes isolated from human fetal epiphyseal cartilage were seeded under mixed conditions into 15-mm-diameter polyglycolic acid (PGA) scaffolds and cultured in recirculation column bioreactors to generate cartilage constructs. After seeding, the cell distributions in thick (4.75 mm) and thin (2.15 mm) PGA disks were nonuniform, with higher cell densities accumulating near the top surfaces. Composite scaffolds were developed by suturing together two thin PGA disks after seeding to manipulate the initial cell distribution before bioreactor culture. The effect of medium flow direction in the bioreactors, including periodic reversal of medium flow, was also investigated. The quality of the tissue-engineered cartilage was assessed after 5 weeks of culture in terms of the tissue wet weight, glycosaminoglycan (GAG), total collagen and collagen type II contents, histological analysis of cell, GAG and collagen distributions, and immunohistochemical analysis of collagen types I and II. Significant enhancement in construct quality was achieved using composite scaffolds compared with single PGA disks. Operation of the bioreactors with periodic medium flow reversal instead of unidirectional flow yielded further improvements in tissue weight and GAG and collagen contents with the composite scaffolds. At harvest, the constructs contained GAG concentrations similar to those measured in ex vivo human adult articular cartilage; however, total collagen and collagen type II levels were substantially lower than those in adult tissue. This study demonstrates that the location of regions of high cell density in the scaffold coupled with application of dynamic bioreactor operating conditions has a significant influence on the quality of tissue-engineered cartilage. 相似文献
15.
Dupont KM Boerckel JD Stevens HY Diab T Kolambkar YM Takahata M Schwarz EM Guldberg RE 《Cell and tissue research》2012,347(3):575-588
Biomaterial scaffolds functionalized to stimulate endogenous repair mechanisms via the incorporation of osteogenic cues offer
a potential alternative to bone grafting for the treatment of large bone defects. We first quantified the ability of a self-complementary
adeno-associated viral vector encoding bone morphogenetic protein 2 (scAAV2.5-BMP2) to enhance human stem cell osteogenic
differentiation in vitro. In two-dimensional culture, scAAV2.5-BMP2-transduced human mesenchymal stem cells (hMSCs) displayed
significant increases in BMP2 production and alkaline phosphatase activity compared with controls. hMSCs and human amniotic-fluid-derived
stem cells (hAFS cells) seeded on scAAV2.5-BMP2-coated three-dimensional porous polymer Poly(ε-caprolactone) (PCL) scaffolds
also displayed significant increases in BMP2 production compared with controls during 12 weeks of culture, although only hMSC-seeded
scaffolds displayed significantly increased mineral formation. PCL scaffolds coated with scAAV2.5-BMP2 were implanted into
critically sized immunocompromised rat femoral defects, both with or without pre-seeding of hMSCs, representing ex vivo and
in vivo gene therapy treatments, respectively. After 12 weeks, defects treated with acellular scAAV2.5-BMP2-coated scaffolds
displayed increased bony bridging and had significantly higher bone ingrowth and mechanical properties compared with controls,
whereas defects treated with scAAV2.5-BMP2 scaffolds pre-seeded with hMSCs failed to display significant differences relative
to controls. When pooled, defect treatment with scAAV2.5-BMP2-coated scaffolds, both with or without inclusion of pre-seeded
hMSCs, led to significant increases in defect mineral formation at all time points and increased mechanical properties compared
with controls. This study thus presents a novel acellular bone-graft-free endogenous repair therapy for orthotopic tissue-engineered
bone regeneration. 相似文献
16.
Magali Cucchiarini Jerome Sohier Karin Mitosch Gunter Kaul David Zurakowski Jeroen M. Bezemer Dieter Kohn Henning Madry 《Central European Journal of Biology》2006,1(1):43-60
Articular cartilage repair might be stimulated by the controlled delivery of therapeutic factors. We tested the hypotheses whether TGF-ß1 can be released from a polymeric scaffold over a prolonged period of time in vitro and whether its transplantation modulates cartilage repair in vivo. Unloaded control or TGF-ß1 poly(ether-ester) copolymeric scaffolds were applied to osteochondral defects in the knee joints of rabbits. In vitro, a cumulative dose of 9 ng TGF-ß1 was released over 4 weeks. In vivo, there were no adverse effects on the synovial membrane. Defects treated with TGF-ß1 scaffolds showed no significant difference in individual parameters of chondrogenesis and in the average cartilage repair score after 3 weeks. There was a trend towards a smaller area (42.5 %) of the repair tissue that stained positive for safranin O in defects receiving TGF-ß1 scaffolds. The data indicate that TGF-ß1 is released from emulsion-coated scaffolds over a prolonged period of time in vitro and that application of these scaffolds does not significantly modulate cartilage repair after 3 weeks in vivo. Future studies need to address the importance of TGF-ß1 dose and release rate to modulate chondrogenesis. 相似文献
17.
En-Rung Chiang Hsiao-Li Ma Jung-Pan Wang Chien-Lin Liu Tain-Hsiung Chen Shih-Chieh Hung 《PloS one》2016,11(2)
Mesenchymal stem cell (MSC)-based therapies may aid in the repair of articular cartilage defects. The purpose of this study was to investigate the effects of intraarticular injection of allogeneic MSCs in an in vivo anterior cruciate ligament transection (ACLT) model of osteoarthritis in rabbits. Allogeneic bone marrow-derived MSCs were isolated and cultured under hypoxia (1% O2). After 8 weeks following ACLT, MSCs suspended in hyaluronic acid (HA) were injected into the knees, and the contralateral knees were injected with HA alone. Additional controls consisted of a sham operation group as well as an untreated osteoarthritis group. The tissues were analyzed by macroscopic examination as well as histologic and immunohistochemical methods at 6 and 12 weeks post-transplantation. At 6 and 12 weeks, the joint surface showed less cartilage loss and surface abrasion after MSC injection as compared to the tissues receiving HA injection alone. Significantly better histological scores and cartilage content were observed with the MSC transplantation. Furthermore, engraftment of allogenic MSCs were evident in surface cartilage. Thus, injection of the allogeneic MSCs reduced the progression of osteoarthritis in vivo. 相似文献
18.
Kazuya Nishizawa Shinji Imai Tomohiro Mimura Mitsuhiko Kubo Susumu Araki Suguru Shioji Yoshinori Takemura Yoshitaka Matsusue 《Cell and tissue research》2010,341(3):371-379
Mesenchymal stromal cells (MSCs), especially those lying close to cartilage defects, are an important cell source for cartilage
regeneration. We hypothesize that a larger number of MSCs might become available, if the bone marrow in the immediate vicinity
of the subchondral bone is stimulated for MSCs in advance of the creation of cartilage defects. A trans-medullary passage-way
reaching the immediate vicinity of the subchondral bone was created 4 days prior to the creation of cartilage defects. In
another setting, basic fibroblast growth factor (bFGF) was administered through the trans-medullary passage-way in order to
augment the stimulation of MSCs. The rabbits were killed at various times after the creation of cartilage defects. Triple
staining of bromodeoxyuridine (BrdU), CD44 and CD45 and histological evaluation were subsequently performed. A considerable
proportion of the proliferating cells were identified as bone-marrow-derived MSCs. Enumeration of BrdU-positive cells demonstrated
that trans-medullary stimulation, especially with bFGF, increased the number of proliferating cells. The histological grading
score of trans-medullary stimulation with bFGF group was superior to that of the other groups. Thus, in-advance stimulation
of the bone marrow effectively increases the number of proliferating cells. The putative progenitor cells for chondrocytes
stimulated thereby are likely to be recruited to the osteochondral defects at the appropriate time, contributing to the repair
of full-thickness articular cartilage defects at the early follow-up time point. 相似文献
19.
Depeng Fang Pan Jin Quanxin Huang Yuan Yang Jinmin Zhao Li Zheng 《Journal of cellular physiology》2019,234(9):15627-15637
The tissue engineering technique using mesenchymal stem cells (MSCs) and scaffolds is promising. Transforming growth factor-β1 (TGF-β1) is generally accepted as an chondrogenic agent, but immunorejection and unexpected side effects, such as tumorigenesis and heterogeneity, limit its clinical application. Autogenous platelet-rich plasma (PRP), marked by low immunogenicity, easy accessibility, and low-cost, may be favorable for cartilage regeneration. In our study, the effect of PRP on engineered cartilage constructed by MSCs and collagen hydrogel in vitro and in vivo was investigated and compared with TGF-β1. The results showed that PRP promoted cell proliferation and gene and protein expressions of chondrogenic markers via the TGF-β/SMAD signaling pathway. Meanwhile, it suppressed the expression of collagen type I, a marker of fibrocartilage. Furthermore, PRP accelerated cartilage regeneration on defects with engineered cartilage, advantageous over TGF-β1, as evaluated by histological analysis and immunohistochemical staining. Our work demonstrates that autogenous PRP may substitute TGF-β1 as a potent and reliable chondrogenic inducer for therapy of cartilage defect. 相似文献
20.
Giovanna Calabrese Raffaella Giuffrida Claudia Fabbi Elisa Figallo Debora Lo Furno Rosario Gulino Cristina Colarossi Francesco Fullone Rosario Giuffrida Rosalba Parenti Lorenzo Memeo Stefano Forte 《PloS one》2016,11(3)
Mesenchymal stem cells (MSCs) play a crucial role in regulating normal skeletal homeostasis and, in case of injury, in bone healing and reestablishment of skeletal integrity. Recent scientific literature is focused on the development of bone regeneration models where MSCs are combined with biomimetic three-dimensional scaffolds able to direct MSC osteogenesis. In this work the osteogenic potential of human MSCs isolated from adipose tissue (hADSCs) has been evaluated in vitro in combination with collagen/Mg doped hydroxyapatite scaffolds. Results demonstrate the high osteogenic potential of hADSCs when cultured in specific differentiation induction medium, as revealed by the Alizarin Red S staining and gene expression profile analysis. In combination with collagen/hydroxyapatite scaffold, hADSCs differentiate into mature osteoblasts even in the absence of specific inducing factors; nevertheless, the supplement of the factors markedly accelerates the osteogenic process, as confirmed by the expression of specific markers of pre-osteoblast and mature osteoblast stages, such as osterix, osteopontin (also known as bone sialoprotein I), osteocalcin and specific markers of extracellular matrix maturation and mineralization stages, such as ALPL and osteonectin. Hence, the present work demonstrates that the scaffold per se is able to induce hADSCs differentiation, while the addition of osteo-inductive factors produces a significant acceleration of the osteogenic process. This observation makes the use of our model potentially interesting in the field of regenerative medicine for the treatment of bone defects. 相似文献