PREPARATION OF ACELLULAR DERMAL MATRIX AS A KIND OF SCAFFOLD FOR CARTILAGE TISSUE ENGINEERING AND ITS BIOCOMPATIBILITY_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

QIHui , JIEYongsheng , CHENLei , JIANGJian , GAOXinsheng ,  SUNLei

Beijing Jishuitan Hospital, Beijing Institute of Traumatology and Orthopaedics, Beijing, 100035, P. R. China;

Corresponding author：

SUNLei, Email: dr_sunlei@263.net

Keywords：

Cartilage tissue engineering; Acellular dermal matrix; Scaffold material; Collagen type Ⅱ

DOI：

10.7507/1002-1892.20140170

Video：

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Objective To study the preparation method of acellular dermal matrix (ADM) for cartilage tissue engineering and analyze its biocompatibility. Methods The dermal tissues of the calf back were harvested, and decelluarized with 0.5% SDS, and the ADM was reconstructed with 0.5% trypsin, cross-linked with formaldehyde, and modified with 0.5% chondroitin sulfate which can promote the proliferation of chondrocytes. And the porosity, cytotoxicity, and biocompatibility were determined. Co-cultured 2nd passage chondrocytes and bone marrow stromal cells in a proportion of 3 to 7 were used as seed cells. The cells were seeded on ADM (experimental group) for 48 hours to observe the cell adhesion. The expressions of mRNA and protein of collagen type Ⅱ were tested by RT-PCR and Western blot methods, respectively. And the expressions were compared between the cells seeded on the scaffold and cultured in monolayer (control group). Results After modification of 0.5% trypsin, the surface of ADM was smooth and had uniform pores; the porosity (85.4%±2.8%) was significantly higher than that without modification (72.8%±5.8%) (t=-4.384, P=0.005). The cell toxicity was grade 1, which accords to the requirements for cartilage tissue engineering scaffolds. With time passing, the number of inflammatory cells decreased after implanted in the back of the rats (P<0.05). The scanning electron microscope observation showed that lots of seed cells adhered to the scaffold, the cells were well stacked, displaying surface microvilli and secretion. The expressions of mRNA and protein of collagen type Ⅱ were not significantly different between experimental and control groups (t=1.265, P=0.235;t=0.935, P=0.372). Conclusion The ADM prepared by acellular treatment, reconstruction, cross-linking, and modification shows perfect characters. And the seed cells maintain chondrogenic phenotype on the scaffold. So it is a proper choice for cartilage tissue engineering.

Citation： QIHui, JIEYongsheng, CHENLei, JIANGJian, GAOXinsheng, SUNLei. PREPARATION OF ACELLULAR DERMAL MATRIX AS A KIND OF SCAFFOLD FOR CARTILAGE TISSUE ENGINEERING AND ITS BIOCOMPATIBILITY. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(6): 768-772. doi: 10.7507/1002-1892.20140170 Copy

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7.	孙磊, 綦惠, 陈磊, 等. 兔软骨细胞与骨髓基质细胞的平面共培养:两者培养比例筛选. 中国组织工程研究与临床康复, 2010, 14(27):4955-4959.
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11.	Livesey SA, Herndon DN, Hollyoak MA, et al. Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis.Transplantation, 1995, 60(1):1-9.
12.	Fosnot J, Kovach SJ 3rd, Serletti JM. Acellular dermal matrix:general principles for the plastic surgeon. Aesthet Surg J, 2011, 31(7 Suppl):5S-12S.
13.	Hochberg M, Chevalier X, Henrotin Y, et al. Symptom and structure modification in osteoarthritis with pharmaceutical-gradechondroitin sulfate:what's the evidence? Curr Med Res Opin, 2013, 29(3):259-267.
14.	Vallières M, du Souich P. Modulation of inflammation by chondroitin sulfate. Osteoarthritis Cartilage, 2010, 18(Suppl 1):S1-6.
15.	Oldershaw RA. Cell sources for the regeneration of articular cartilage:the past, the horizon and the future. Int J Exp Pathol, 2012, 93(6):389-400.
16.	Ma D, Ren L, Liu Y, et al. Engineering scaffold-free bone tissue using bone marrow stromal cell sheets. J Orthop Res, 2010, 28(5):697-702.

1. Marcacci M, Filardo G, Kon E. Treatment of cartilage lesions:what works and why? Injury, 2013, 44 Suppl 1:S11-15.
2. Nukavarapu SP, Dorcemus DL. Osteochondral tissue engineering:current strategies and challenges. Biotechnol Adv, 2013, 31(5):706-721.
3. Kon E, Filardo G, Di Martino A, et al. ACI and MACI. J Knee Surg, 2012, 25(1):17-22.
4. Getgood A, Brooks R, Fortier L, et al. Articular cartilage tissue engineering:today's research, tomorrow's practice? J Bone Joint Surg (Br), 2009, 91(5):565-576.
5. Vinatier C, Mrugala D, Jorgensen C, et al. Cartilage engineering:a crucial combination of cells, biomaterials and biofactors. Trends Biotechnol, 2009, 27(5):307-314.
6. Simon TM, Jackson DW. Articular cartilage:injury pathways and treatment options. Sports Med Arthrosc, 2006, 14(3):146-154.
7. 孙磊, 綦惠, 陈磊, 等. 兔软骨细胞与骨髓基质细胞的平面共培养:两者培养比例筛选. 中国组织工程研究与临床康复, 2010, 14(27):4955-4959.
8. Chajra H, Rousseau CF, Cortial D, et al. Collagen-based biomaterials and cartilage engineering. Application to osteochondral defects. Biomed Mater Eng, 2008, 18(1 Suppl):S₃3-45.
9. Stone KR, Steadman JR, Rodkey WG, et al. Regeneration of meniscal cartilage with use of a collagen scaffold. J Bone Joint Surg (Am), 1997, 79(12):1770-1777.
10. Heck EL, Bergstresser PR, Baxter CR. Composite skin graft:frozen dermal allografts support the engraftment and expansion of autologous epidermis. J Trauma, 1985, 25(2):106-112.
11. Livesey SA, Herndon DN, Hollyoak MA, et al. Transplanted acellular allograft dermal matrix. Potential as a template for the reconstruction of viable dermis.Transplantation, 1995, 60(1):1-9.
12. Fosnot J, Kovach SJ 3rd, Serletti JM. Acellular dermal matrix:general principles for the plastic surgeon. Aesthet Surg J, 2011, 31(7 Suppl):5S-12S.
13. Hochberg M, Chevalier X, Henrotin Y, et al. Symptom and structure modification in osteoarthritis with pharmaceutical-gradechondroitin sulfate:what's the evidence? Curr Med Res Opin, 2013, 29(3):259-267.
14. Vallières M, du Souich P. Modulation of inflammation by chondroitin sulfate. Osteoarthritis Cartilage, 2010, 18(Suppl 1):S1-6.
15. Oldershaw RA. Cell sources for the regeneration of articular cartilage:the past, the horizon and the future. Int J Exp Pathol, 2012, 93(6):389-400.
16. Ma D, Ren L, Liu Y, et al. Engineering scaffold-free bone tissue using bone marrow stromal cell sheets. J Orthop Res, 2010, 28(5):697-702.

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PREPARATION OF ACELLULAR DERMAL MATRIX AS A KIND OF SCAFFOLD FOR CARTILAGE TISSUE ENGINEERING AND ITS BIOCOMPATIBILITY

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