Chondrocyte differentiation of dog bone marrow mesenchymal stem cells co-cultured with Bio-gide collagen membrane in vitro

doi:10.3969/j.issn.2095-4344.2013.29.004

Abstract

Abstract:

BACKGROUND: Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method.
OBJECTIVE: To explore the feasibility of Bio-gide collagen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes.
METHODS: Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cell surface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). Cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. Cells underwent type Ⅱ collagen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide collagen membrane were observed under an inverted phase contrast microscope and scanning electron microscope.
RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. Cells grew well
and had strong amplified ability, and successfully differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide collagen membrane, showing a tendency of multi-layer growth. Cells and Bio-gide collagen membrane seem to blend into an integrant part. Cell processes appeared and connected each other and gradually wrapped the Bio-gide collagen membrane, with the presence of obvious cell matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide collagen membrane.

Key words: biomaterials, tissue-engineered cartilage bone materials, membrane biomaterials, bone marrow mesenchymal stem cells, Bio-gide collagen membrane, tissue engineering, osteonecrosis of the femoral head, cartilage defects, seed cells, National Natural Science Foundation of China

CLC Number:

R318

Xie Hui, Yang Fei, Zhao De-wei, Wang Ben-jie, Cui Da-ping, Wang Wei, Huang Shi-bo. Chondrocyte differentiation of dog bone marrow mesenchymal stem cells co-cultured with Bio-gide collagen membrane in vitro[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(29): 5282-5289.

Figures/Tables 5

References

[1] Zhao D,Wang B,Guo L,et al.Will a Vascularized Greater Trochanter Graft Preserve the Necrotic Femoral Head? Clin Orthop Relat Res.2010;468: 1316-1324.

[2] Li Y,Han R,Geng C,et al. A new osteonecrosis animal model of the femoral head induced by microwave heating and repaired with tissue engineered bone. Int Orthop.2009;33(2)：573-580.

[3] Peng J,Wen C,Wang A,et al. Micro-CT-based bone ceramic scaffolding and its performance after seeding with mesenchymal stem cells for repair of load-bearing bone defect in canine femoral head.J Biomed Mater Res B Appl Biomater.2011;96(2):316-325.

[4] Hartman GA,Arnold RM,Mills MP,et al.Clinical and histologic evaluation of anorganic bovine bone collagen with or without a collagen barrier.Int J Periodontics Restorative Dent. 2004; 24(2):127-135.

[5] Nevo Z,Robinson D,Horowitz S,et al.Collagen membranes at immediate implants: a histomorphometric study in dogs.Clin Oral Implants Res.Ceu Transplantation.1998;7(1): 63-70.

[6] Wakitani S, Goto T, Young RG,et al.Repari of large full-thickness articular cartilage defects with allograft artichlar chondrocytes embedded in a collagengel.Tissue Eng.1998; 4(4): 429-444.

[7] Marc L,Camelo M, Samuel E, et al. Evaluation of periodontal regeneration following grafting intrabony defects with bio-oss collagen:a human histologic report. Int J Periedonties Restorative Dent.2003;23(1):9-17.

[8] 张志兴,詹暶,赵欣,等.不同浓度的骨髓基质细胞裸鼠体内成骨能力的研究[J].口腔医学研究,2009,25(6):720-722.

[9] 张向荣,郭光华,刘德伍,等. 人骨髓间充质干细胞的分离培养及BrdU标记鉴定[J].中国组织工程研究与临床康复,2009,13(19): 3618-3622.

[10] Barone FC,Arvin B,White RF,et al.T umor necrosis factor-alpha. a mediator necrosis factor-alpha.a mediator of focal ischemic brain injury.Storke.1997;28(6):1233-1244.

[11] Lee HS,Huang GT,Chiang H,et al. Multi-potential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. Stem cells.2003;21: 190-199.

[12] Lin Z,Willers C,Xu J,et al. The chondrocyte: biology and clinical application.Tissue Eng. 2006;12(7):1971-1984.

[13] 潘思年,杜敏联,马华梅,等. 大鼠胫骨生长板软骨细胞的体外培养及鉴定[J].中山大学学报:医学科学版,2009,30(45):1-5.

[14] 赵德伟,胡永成. 成人股骨头坏死诊疗标准专家共识(2012年版)[J].中华关节外科杂志,2012,6(3):479-484.

[15] Sekiya JK,Ruch DS,Hunter DM,et al. Hip arthroscopy in staging avascular necrosis of the femoral head. South Orthop Assoc.2000;9(4):3-9.

[16] Urbaniak JR,Coogan PG,Gunneson EB,et al.Treatment of osteonecrosis of the femoral head with free vascularized fibular grafting: a long-term follow-up study of on e hundred and three hips. J Bone Joint Surg Am.1999;577(6): 681- 694.

[17] Mats Brittberg. 细胞载体作为软骨修复中的下一代细胞疗法[J].中华关节外科杂志,2011,5(6):796-812.

[18] Abelow SP,Guillen P,Ramos T. Arthroscopic technique for matrix-induced autologous chondrocyte implantation for the treatment of large chondral defects in the knee and ankle. Oper Tech Orthop.2006; 16: 257-261.

[19] Amin AA,Bartlett W,Gooding CR,et al. The use of autologous chondrocyte implantation following and combined with anterior cruciate ligament reconstruction.Int Orthop.2006; 30(1):48-53.

[20] Bachmann G,Basad E,Lommel D,et al. MRT in the follow-up of matrix-guided autologous chondrocyte implantation ( MACI ) and microfracture. Radiologe.2004; 44(8): 773-782.

[21] Bartlett W,Gooding CR,Carrington RWJ,et al. Autologous chondrocyte implantation at the knee using a bilayer collagen membrane with bone graft. A preliminary report. J Bone Joint Surg Br.2005; 87: 330-332.

[22] Bartlett W,Krishnan SP,Skinner JA,et al. Collagen-covered versus matrix-induced autologous chondrocyte implantation for osteochondral defects of the knee. A comparison of tourniquet times. Eur J Orthop Surg Traumatol.2006; 16: 315-317.

[23] Bartlett W,Skinner JA,Gooding CR,et al. Autologous chondrocyte implantation versus matrix-induced autologous chondrocyte implantation for osteochondral defects of the knee: a prospective,randomised study. J Bone Joint Surg Br. 2005; 87(5): 640-645.

[24] Behrens P,Bitter T,Kurz B,et al. Matrix-associated autologous chondrocyte transplantation /implantation ( MACT/MACI)-5-year follow-up. Knee.2006; 13(3): 194-202.

[25] Cherubino P,Grassi FA,Bulgheroni P,et al. Autologous chondrocyte implantation using a bilayer collagen membrane: a preliminary report. J Orthop Surg.2003; 11(1): 10 -15.

[26] D’Anchise R,Manta N,Prospero E,et al. Autologous implantation of chondrocytes on a solid collagen scaffold: clinical and histological outcomes after two years of follow-up. J Orthopaed Traumatol.2005; 6: 36-43.

[27] Macarini L,Murrone M,Marini S,et al. Aspects of magnetic resonance in the surgical treatment of osteochondral lesions of the knee. Radiol Med.2003; 106(1-2) : 74-86.

[28] Marlovits S,Striessnig G,Kutscha-Lissberg F,et al. Early postoperative adherence of matrix-induced autologous chondrocyte implantation for the treatment of full-thickness cartilage defects of the femoral condyle. Knee Surg Sports Traumatol Arthrosc.2005; 13(6) : 451-457.

[29] Marlovits S,Striessnig G,Resinger CT,et al. Definition of pertinent parameters for the evaluation of articular cartilage repair tissue with high-resolution magnetic resonance imaging. Eur J Radiol.2004; 52(3):310-319.

[30] Ronga M,Grassi FA,Bulgheroni P. Arthroscopic autologous chondrocyte implantation for the treatment of a chondral defect in the tibial plateau of the knee. Arthroscopy.2004; 20(1):79-84.

[31] Ronga M,Grassi FA,Manelli A,et al. Tissue engineering techniques for the treatment of a complex knee injury. Arthroscopy.2006; 22(5):576.e1-576 e3.

[32] Ronga M,Grassi FA,Montoli C,et al. Treatment of deep cartilage defects of the ankle with matrix-induced autologous chondrocyte implantation ( MACI).Foot Ankle Surg.2005; 11: 29-33.

[33] Trattnig S,Ba-Ssalamah A,Pinker K,et al. Matrix-based autologous chondrocyte implantation for cartilage repair: noninvasive monitoring by high-resolution magnetic resonance imaging. Magn Reson Imaging.2005; 23(7):779- 787.

[34] Zhang Z,Ye Q,Yang Z,et al. Matrix-induced autologous chondrocyte implantation for treatment of chondral defects of knee: a preliminary report. J Musculoskel Res.2006; 10: 95-101.

[35] Zheng MH,Willers C,Kirilak L,et al. Matrix-induced autologous chondrocyte implantation ( MACI) : biological and histological assessment. Tissue Eng.2007; 13(4): 737-746.

[36] 荆志成.心脏胶原网络及其病理学意义[J].中国循环杂志,1998, 13(3):187-188.

[37] 贺冬冬,曾令员,向川,等.Ⅰ/Ⅲ型胶原膜联合自体骨髓间充质干细胞修复兔膝关节软骨缺损[J].中国组织工程研究,2012,16(38): 7031-7036.

[38] Vander Kraan PM , Buma P, Van Kuppevelt T, et al . Interaction of chondrocytes, extracellular matrix and growth factors :relevance for articular cartilage tissue engineering. Osteoarthritis Cartilage.2002; 10(8): 631-637.