Molecular and immunohistological characterization of human cartilage two years following autologous cell transplantation

Brunella Grigolo, Livia Roseti, Luciana De Franceschi, Anna Piacentini, Luca Cattini, Massimiliano Manfredini, Riccardo Faccini, Andrea Facchini

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)46-57
Number of pages12
JournalJournal of Bone and Joint Surgery - Series A
Volume87
Issue number1
DOIs
Publication statusPublished - Jan 2005

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Autologous Transplantation
Cell Transplantation
Chondrocytes
Early Growth Response Protein 1
Collagen Type II
Cartilage
Cathepsin B
Collagen Type I
Collagen Type X
Aggrecans
Messenger RNA
Biopsy
Biochemical Phenomena
Genetic Phenomena
Proteins
Extracellular Matrix Proteins
Articular Cartilage
Reverse Transcriptase Polymerase Chain Reaction
Paraffin
Extracellular Matrix

ASJC Scopus subject areas

  • Surgery
  • Orthopedics and Sports Medicine

Cite this

Molecular and immunohistological characterization of human cartilage two years following autologous cell transplantation. / Grigolo, Brunella; Roseti, Livia; De Franceschi, Luciana; Piacentini, Anna; Cattini, Luca; Manfredini, Massimiliano; Faccini, Riccardo; Facchini, Andrea.

In: Journal of Bone and Joint Surgery - Series A, Vol. 87, No. 1, 01.2005, p. 46-57.

Research output: Contribution to journalArticle

Grigolo, Brunella ; Roseti, Livia ; De Franceschi, Luciana ; Piacentini, Anna ; Cattini, Luca ; Manfredini, Massimiliano ; Faccini, Riccardo ; Facchini, Andrea. / Molecular and immunohistological characterization of human cartilage two years following autologous cell transplantation. In: Journal of Bone and Joint Surgery - Series A. 2005 ; Vol. 87, No. 1. pp. 46-57.
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abstract = "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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AU - Grigolo, Brunella

AU - Roseti, Livia

AU - De Franceschi, Luciana

AU - Piacentini, Anna

AU - Cattini, Luca

AU - Manfredini, Massimiliano

AU - Faccini, Riccardo

AU - Facchini, Andrea

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N2 - 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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