Generation of a Scaffold Free Cartilage-like Implant from a Small Amount of Starting Material

Overview

Journal J Cell Mol Med

Specialties Cell Biology
Molecular Biology

Date 2006 Jun 27

PMID 16796813

Citations 6

Authors

M J Stoddart

L Ettinger

H J Hauselmann

Affiliations

Soon will be listed here.

Abstract

Introduction: An autologous cellular based treatment of a traumatic cartilage injury requires a procedure whereby a biopsy of healthy cartilage is removed from the patient and the cells isolated and expanded by monolayer passage. This increases the cell number to required levels but also leads to a de-differentiation of the cells. We aim to produce a scaffold-free, de-novo implant from a biopsy of cartilage.

Methods: Bovine chondrocytes were isolated from a small biopsy and expanded. The chondrocytic phenotype of the monolayer expanded cells was recovered during a period of culture in alginate and the effect of factors such as IGF1, TFGbeta1 and dexamethasone was investigated.

Results: During the alginate culture period a pre-treatment with IGF1 and dexamethasone was shown to have little effect. IGF1 however increased the glycosaminoglycan/DNA (GAG/DNA) content on day 14 to 84.95+/-5 ng/ng compared with 37.3+/-1.8 ng/ng in the controls (P<0.001). 35S labeling demonstrated an increased GAG synthesis in the presence of IGF1 (P<0.001). IGF1 also induced a increase of DNA content 1383+/-314 ng/bead compared to 512+/-19 ng/bead in the controls (P<0.001). The cells were released from the alginate and cultured in a silicon mould for a further 14 days to obtain a three dimensional implant. Releasing the cells from the alginate and casting in a mould produced an implant of defined shape which contained no foreign material. After 31 days of culture the implants contained 152.4+/-13.14 ng/ng GAG/DNA and 42.93+/-10.23 ng/ng collagen II.

Discussion: We believe alginate released chondrocytes provide a real alternative to artificial scaffolds.

Citing Articles

Metabolomic Profiling and Characterization of a Novel 3D Culture System for Studying Chondrocyte Mechanotransduction.

Brahmachary P, Brahmachary P, Erdogan E, Erdogan A, Myers E, Myers E bioRxiv. 2024; .

PMID: 38915493 PMC: 11195103. DOI: 10.1101/2024.06.10.598340.

Metabolomic Profiling and Mechanotransduction of Single Chondrocytes Encapsulated in Alginate Microgels.

Fredrikson J, Brahmachary P, Erdogan A, Archambault Z, Wilking J, June R Cells. 2022; 11(5).

PMID: 35269522 PMC: 8909502. DOI: 10.3390/cells11050900.

A Concise Review on the Use of Mesenchymal Stem Cells in Cell Sheet-Based Tissue Engineering with Special Emphasis on Bone Tissue Regeneration.

Yorukoglu A, Kiter A, Akkaya S, Lale Satiroglu-Tufan N, Tufan A Stem Cells Int. 2017; 2017:2374161.

PMID: 29230248 PMC: 5694585. DOI: 10.1155/2017/2374161.

The effect of dexamethasone and triiodothyronine on terminal differentiation of primary bovine chondrocytes and chondrogenically differentiated mesenchymal stem cells.

Randau T, Schildberg F, Alini M, Wimmer M, Haddouti E, Gravius S PLoS One. 2013; 8(8):e72973.

PMID: 23977373 PMC: 3745539. DOI: 10.1371/journal.pone.0072973.

Shaped, stratified, scaffold-free grafts for articular cartilage defects.

Han E, Bae W, Hsieh-Bonassera N, Wong V, Schumacher B, Gortz S Clin Orthop Relat Res. 2008; 466(8):1912-20.

PMID: 18506565 PMC: 2584257. DOI: 10.1007/s11999-008-0291-7.

References

Homicz M, Chia S, Schumacher B, Masuda K, Thonar E, Sah R . Human septal chondrocyte redifferentiation in alginate, polyglycolic acid scaffold, and monolayer culture. Laryngoscope. 2003; 113(1):25-32. DOI: 10.1097/00005537-200301000-00005. View

AYDELOTTE M, Kuettner K . Differences between sub-populations of cultured bovine articular chondrocytes. I. Morphology and cartilage matrix production. Connect Tissue Res. 1988; 18(3):205-22. DOI: 10.3109/03008208809016808. View

Alsalameh S, Amin R, Gemba T, Lotz M . Identification of mesenchymal progenitor cells in normal and osteoarthritic human articular cartilage. Arthritis Rheum. 2004; 50(5):1522-32. DOI: 10.1002/art.20269. View

Jakob M, Demarteau O, Schafer D, Hintermann B, Dick W, Heberer M . Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro. J Cell Biochem. 2001; 81(2):368-77. DOI: 10.1002/1097-4644(20010501)81:2<368::aid-jcb1051>3.0.co;2-j. View

Bjornsson S . Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue. Anal Biochem. 1993; 210(2):282-91. DOI: 10.1006/abio.1993.1197. View

Paige K, Cima L, Yaremchuk M, Schloo B, Vacanti J, Vacanti C . De novo cartilage generation using calcium alginate-chondrocyte constructs. Plast Reconstr Surg. 1996; 97(1):168-78; discussion 179-80. DOI: 10.1097/00006534-199601000-00027. View

Benya P, SHAFFER J . Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982; 30(1):215-24. DOI: 10.1016/0092-8674(82)90027-7. View

Barbero A, Ploegert S, Heberer M, Martin I . Plasticity of clonal populations of dedifferentiated adult human articular chondrocytes. Arthritis Rheum. 2003; 48(5):1315-25. DOI: 10.1002/art.10950. View

Binette F, McQuaid D, Haudenschild D, Yaeger P, McPherson J, Tubo R . Expression of a stable articular cartilage phenotype without evidence of hypertrophy by adult human articular chondrocytes in vitro. J Orthop Res. 1998; 16(2):207-16. DOI: 10.1002/jor.1100160208. View

10.

Li W, Tuli R, Okafor C, Derfoul A, Danielson K, Hall D . A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. Biomaterials. 2004; 26(6):599-609. DOI: 10.1016/j.biomaterials.2004.03.005. View

11.

Pascher A, Palmer G, Steinert A, Oligino T, Gouze E, Gouze J . Gene delivery to cartilage defects using coagulated bone marrow aspirate. Gene Ther. 2004; 11(2):133-41. DOI: 10.1038/sj.gt.3302155. View

12.

Bonaventure J, Kadhom N, COHEN-SOLAL L, Ng K, Bourguignon J, Lasselin C . Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. Exp Cell Res. 1994; 212(1):97-104. DOI: 10.1006/excr.1994.1123. View

13.

Mizuno S, Allemann F, Glowacki J . Effects of medium perfusion on matrix production by bovine chondrocytes in three-dimensional collagen sponges. J Biomed Mater Res. 2001; 56(3):368-75. DOI: 10.1002/1097-4636(20010905)56:3<368::aid-jbm1105>3.0.co;2-v. View

14.

Grigolo B, Roseti L, Fiorini M, Fini M, Giavaresi G, Aldini N . Transplantation of chondrocytes seeded on a hyaluronan derivative (hyaff-11) into cartilage defects in rabbits. Biomaterials. 2001; 22(17):2417-24. DOI: 10.1016/s0142-9612(00)00429-4. View

15.

Schaefer D, Martin I, Jundt G, Seidel J, Heberer M, Grodzinsky A . Tissue-engineered composites for the repair of large osteochondral defects. Arthritis Rheum. 2002; 46(9):2524-34. DOI: 10.1002/art.10493. View

16.

Anderer U, Libera J . In vitro engineering of human autogenous cartilage. J Bone Miner Res. 2002; 17(8):1420-9. DOI: 10.1359/jbmr.2002.17.8.1420. View

17.

Hedbom E, Ettinger L, Hauselmann H . Culture of articular chondrocytes in alginate gel--a means to generate cartilage-like implantable tissue. Osteoarthritis Cartilage. 2001; 9 Suppl A:S123-30. View

18.

Yang L, Korom S, Welti M, Hoerstrup S, Zund G, Jung F . Tissue engineered cartilage generated from human trachea using DegraPol scaffold. Eur J Cardiothorac Surg. 2003; 24(2):201-7. DOI: 10.1016/s1010-7940(03)00263-x. View

19.

Chia S, Schumacher B, Klein T, Thonar E, Masuda K, Sah R . Tissue-engineered human nasal septal cartilage using the alginate-recovered-chondrocyte method. Laryngoscope. 2004; 114(1):38-45. DOI: 10.1097/00005537-200401000-00006. View

20.

Hauselmann H, Masuda K, Hunziker E, Neidhart M, Mok S, Michel B . Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage. Am J Physiol. 1996; 271(3 Pt 1):C742-52. DOI: 10.1152/ajpcell.1996.271.3.C742. View