The Journal of Bone and Joint Surgery (American). 2005;87:46-57.
doi:10.2106/JBJS.C.01685
© 2005 The Journal of Bone and Joint Surgery, Inc.
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Molecular and Immunohistological Characterization of Human Cartilage Two Years Following Autologous Cell Transplantation

Brunella Grigolo, PhD1, Livia Roseti, PhD1, Luciana De Franceschi, PhD1, Anna Piacentini, PhD1, Luca Cattini, BSc1, Massimiliano Manfredini, MD2, Riccardo Faccini, MD2 and Andrea Facchini, MD3

1 Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla Putti, Istituti Ortopedici Rizzoli, Via di Barbiano 1/10, 40136 Bologna, Italy. E-mail address for B. Grigolo: grigolob{at}alma.unibo.it
2 Divisione di Ortopedia e Traumatologia, Ospedale Del Delta, Via Valle Oppio 2, 44023 Lagosanto, Ferrara, Italy
3 Dipartimento di Medicina Interna e Gastroenterologia, Università di Bologna, Istituti Ortopedici Rizzoli, Via di Barbiano 1/10, 40136 Bologna, Italy

Investigation performed at Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla Putti, Istituti Ortopedici Rizzoli, Bologna, Italy

In support of their research or preparation of this manuscript, one or more of the authors received grants or outside funding from Genzyme Biosurgery, Cambridge, Massachusetts. None of the authors received payments or other benefits or a commitment or agreement to provide such benefits from a commercial entity. No commercial entity paid or directed, or agreed to pay or direct, any benefits to any research fund, foundation, educational institution, or other charitable or nonprofit organization with which the authors are affiliated or associated.

Background: There are only a few studies concerning the cellular, biochemical, and genetic processes that occur during the remodeling of graft tissue after autologous chondrocyte transplantation. The purpose of the present study was to quantify the expression of genes encoding extracellular matrix proteins and regulatory factors that are essential for cell differentiation in cartilage biopsy specimens from patients who had this treatment two years previously.

Methods: Two cartilage biopsy specimens from each of four patients who had been treated with autologous chondrocyte transplantation and from two multiorgan donors were used. Real-time reverse transcriptase-polymerase chain reaction analysis was performed to evaluate the expression of types I, II, and X collagen; aggrecan; cathepsin B; and early growth response protein-1 (Egr-1) and Sry-type high-mobility-group box transcription factor-9 (Sox-9) mRNAs. Immunohistochemical analysis for matrix proteins and regulatory proteins was carried out on paraffin-embedded sections.

Results: Type-I collagen mRNA was expressed in all of the samples evaluated. Type-II collagen was present in autologous chondrocyte transplantation samples but at lower levels than in the controls. Type-X collagen messenger was undetectable. Aggrecan mRNA was present in all of the samples at lower levels than in the controls, while cathepsin-B messenger levels were higher and Egr-1 and Sox-9 mRNAs were expressed at lower levels. The immunohistochemical analysis showed slight positivity for type-I collagen in all of the sections. Type-II collagen was found in all of the samples with positivity confined inside the cells, while the controls displayed a positivity that was diffuse in the extracellular matrix. Cathepsin B was slightly positive in all of the samples, while the controls were negative. Egr-1 protein was particularly evident in the areas negative for type-II collagen. Sox-9 was positive in all samples, with evident localization in the superficial and middle layers.

Conclusions: In biopsy specimens from autologous chondrocyte transplantation tissue at two years, there is evidence of the formation of new tissue, which displays varying degrees of organization with some fibrous and fibrocartilaginous features. Long-term follow-up investigations are needed to verify whether, once all of the remodeling processes are completed, the newly formed tissue will acquire the more typical features of articular cartilage.

Clinical Relevance: Our data provide important further information about the nature of the new cartilage tissue formed two years after autologous chondrocyte transplantation, allowing a better interpretation of the clinical and histological findings.


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