rAAV-Mediated sox9 Overexpression Improves the Repair of Osteochondral Defects in a Clinically Relevant Large Animal Model Over Time In Vivo and Reduces Perifocal Osteoarthritic Changes

2021 ◽  
pp. 036354652110494
Author(s):  
Cliff Lange ◽  
Henning Madry ◽  
Jagadeesh K. Venkatesan ◽  
Gertrud Schmitt ◽  
Susanne Speicher-Mentges ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Gene Transfer ◽  
Subchondral Bone ◽  
Cartilage Defects ◽  
Large Animal ◽  
Type I ◽  
Lacz Gene ◽  
Osteochondral Defects ◽  
Osteochondral Repair ◽  
Over Time

Background: Gene transfer of the transcription factor SOX9 with clinically adapted recombinant adeno-associated virus (rAAV) vectors offers a powerful tool to durably enhance the repair process at sites of osteochondral injuries and counteract the development of perifocal osteoarthritis (OA) in the adjacent articular cartilage. Purpose: To examine the ability of an rAAV sox9 construct to improve the repair of focal osteochondral defects and oppose perifocal OA development over time in a large translational model relative to control gene transfer. Study Design: Controlled laboratory study. Methods: Standardized osteochondral defects created in the knee joints of adult sheep were treated with rAAV-FLAG-h sox9 relative to control (reporter) rAAV- lacZ gene transfer. Osteochondral repair and degenerative changes in the adjacent cartilage were monitored using macroscopic, histological, immunohistological, and biochemical evaluations after 6 months. The microarchitecture of the subchondral bone was assessed by micro–computed tomography. Results: Effective, prolonged sox9 overexpression via rAAV was significantly achieved in the defects after 6 months versus rAAV- lacZ treatment. The application of rAAV-FLAG-h sox9 improved the individual parameters of defect filling, matrix staining, cellular morphology, defect architecture, surface architecture, subchondral bone, and tidemark as well as the overall score of cartilage repair in the defects compared with rAAV- lacZ. The overexpression of sox9 led to higher levels of proteoglycan production, stronger type II collagen deposition, and reduced type I collagen immunoreactivity in the sox9- versus lacZ-treated defects, together with decreased cell densities and DNA content. rAAV-FLAG-h sox9 enhanced semiquantitative histological subchondral bone repair, while the microstructure of the incompletely restored subchondral bone in the sox9 defects was not different from that in the lacZ defects. The articular cartilage adjacent to the sox9-treated defects showed reduced histological signs of perifocal OA changes versus rAAV- lacZ. Conclusion: rAAV-mediated sox9 gene transfer enhanced osteochondral repair in sheep after 6 months and reduced perifocal OA changes. These results underline the potential of rAAV-FLAG-h sox9 as a therapeutic tool to treat cartilage defects and afford protection against OA. Clinical Relevance: The delivery of therapeutic rAAV sox9 to sites of focal injuries may offer a novel, convenient tool to enhance the repair of osteochondral defects involving both the articular cartilage and the underlying subchondral bone and provide a protective role by reducing the extent of perifocal OA.

rAAV-Mediated Human FGF-2 Gene Therapy Enhances Osteochondral Repair in a Clinically Relevant Large Animal Model Over Time In Vivo

2021 ◽  
Vol 49 (4) ◽  
pp. 958-969
Author(s):  
Yannik P. Morscheid ◽  
Jagadeesh K. Venkatesan ◽  
Gertrud Schmitt ◽  
Patrick Orth ◽  
David Zurakowski ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Animal Model ◽  
Gene Transfer ◽  
Subchondral Bone ◽  
Type I ◽  
Osteochondral Defects ◽  
Osteochondral Repair ◽  
The Individual ◽  
Over Time

Background: Osteochondral defects, if left untreated, do not heal and can potentially progress toward osteoarthritis. Direct gene transfer of basic fibroblast growth factor 2 (FGF-2) with the clinically adapted recombinant adeno-associated viral (rAAV) vectors is a powerful tool to durably activate osteochondral repair processes. Purpose: To examine the ability of an rAAV-FGF-2 construct to target the healing processes of focal osteochondral injury over time in a large translational model in vivo versus a control gene transfer condition. Study Design: Controlled laboratory study. Methods: Standardized osteochondral defects created in the knee joints of adult sheep were treated with an rAAV human FGF-2 (hFGF-2) vector by direct administration into the defect relative to control (reporter) rAAV- lacZ gene transfer. Osteochondral repair was monitored using macroscopic, histological, immunohistological, and biochemical methods and by micro–computed tomography after 6 months. Results: Effective, localized prolonged FGF-2 overexpression was achieved for 6 months in vivo relative to the control condition without undesirable leakage of the vectors outside the defects. Such rAAV-mediated hFGF-2 overexpression significantly increased the individual histological parameter “percentage of new subchondral bone” versus lacZ treatment, reflected in a volume of mineralized bone per unit volume of the subchondral bone plate that was equal to a normal osteochondral unit. Also, rAAV-FGF-2 significantly improved the individual histological parameters “defect filling,”“matrix staining,” and “cellular morphology” and the overall cartilage repair score versus the lacZ treatment and led to significantly higher cell densities and significantly higher type II collagen deposition versus lacZ treatment. Likewise, rAAV-FGF-2 significantly decreased type I collagen expression within the cartilaginous repair tissue. Conclusion: The current work shows the potential of direct rAAV-mediated FGF-2 gene therapy to enhance osteochondral repair in a large, clinically relevant animal model over time in vivo. Clinical Relevance: Delivery of therapeutic (hFGF-2) rAAV vectors in sites of focal injury may offer novel, convenient tools to enhance osteochondral repair in the near future.

scholarly journals The Patello-Femoral Joint Degeneration and the Shape of the Patella in the Population Needing an Arthroscopic Procedure

Medicina ◽  
2018 ◽  
Vol 54 (2) ◽  
pp. 21 ◽  
Author(s):  
Rimtautas Gudas ◽  
Laimonas Šiupšinskas ◽  
Agnė Gudaitė ◽  
Vladas Vansevičius ◽  
Edgaras Stankevičius ◽  
...  
Keyword(s):  
Physical Activity ◽  
Articular Cartilage ◽  
Knee Joint ◽  
Cartilage Defects ◽  
Type I ◽  
Physically Active ◽  
Group Iii ◽  
Type Iii ◽  
Group I ◽  
Arthroscopic Procedures

Background: the main goal of the study was to investigate the prevalence of the articular cartilage defects (ACD) in the patellofemoral (PF) region of the knee joint based on the anatomical shapes of patella and its impact on the level of physical activity in the population needing arthroscopic procedures for all types of pathologies in the knee. Methods: The articular cartilage status of the PF region was obtained from 1098 arthroscopic procedures of the knee joint. The ACD were correlated to Wiberg’s shape of the patella and classified according to the degree, size and depth of the ACD in the PF region using the ICRS (International Cartilage Repair Society) system: group I consisting of patients with Wiberg type I shape (W1), group II—patients with Wiberg type II shape (W2) and group III—patients with Wiberg type III shape (W3). The Tegner physical activity scale was used to evaluate the physical activity of the patients. Results: The mean of ACD size (PF region) in the W3 group was 3.10 ± 0.99 cm2, which was a statistically significantly larger area in comparison with the W1 (1.90 ± 0.63 cm2; p < 0.0000) and W2 (1.95 ± 0.71 cm2; p < 0.0000). The patients from the W3 group (mean 3.10 ± 0.99) were less physically active (<4 Tegner) compared to the W2 group (mean of 4.48 ± 0.88; p = 0.004) and W1 group (mean of 4.55 ± 0.72; p = 0.002). Conclusions: The patients with the Wiberg type III patella shape had a higher incidence and larger size of ACD in the PF of the knee compared to the groups of Wiberg type I and II. Wiberg III patients with a lower level of physical activity had a larger size of ACD in the PF joint.

A CURRENT REVIEW ON THE BIOLOGY AND TREATMENT OF ARTICULAR CARTILAGE DEFECTS (PART I & PART II)

2003 ◽  
Vol 07 (03n04) ◽  
pp. 157-181 ◽  
Author(s):  
Craig Willers ◽  
David J. Wood ◽  
Ming H. Zheng
Keyword(s):  
Articular Cartilage ◽  
Subchondral Bone ◽  
18Th Century ◽  
Later Life ◽  
Fibrin Clot ◽  
Joint Disease ◽  
Cartilage Defects ◽  
Adult Males ◽  
Chondrocyte Implantation ◽  
Osteochondral Injury

Osteochondral injury occurs predominantly in physically active young adult males. Injury to the articular cartilage and/or subchondral bone may not only cause acute joint disease resulting in osseous intracapsular (synovitis) or extracapsular pain, but may also act to spawn arthritic conditions in later life. Since the 18th century, such injury has proven difficult to treat clinically, and much therapy has been essentially palliative. Past treatments such as abrasion arthroplasty, drilling, microfracture and arthroscopic lavage have been useful in removing articular debris and promoting the formation of the fibrin clot used in most native repair mechanisms. However, the limitation of these techniques is their inability to restore the damaged cartilage and subchondral bone to their normal tissue architecture. Recent developments in tissue engineering have concentrated on the utilization of autologous chondrocyte implantation, biomaterials and growth factors to promote the regeneration of biomechanically superior hyaline articular cartilage. This paper reviews the etiology, repair biology and therapeutic techniques of cartilage and/or osteochondral injury over the previous decades, and attempts to provide insight into interesting new research directions which offer much potential for improved treatment of these troublesome lesions.

scholarly journals Ex vivo gene transfer to chondrocytes in full-thickness articular cartilage defects: a feasibility study

1997 ◽  
Vol 5 (2) ◽  
pp. 139-143 ◽  
Author(s):  
Richard Kang ◽  
Takashi Marui ◽  
Steven C. Ghivizzani ◽  
Ioana M. Nita ◽  
Helga I. Georgescu ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Gene Transfer ◽  
Feasibility Study ◽  
Ex Vivo ◽  
Cartilage Defects ◽  
Full Thickness ◽  
Ex Vivo Gene Transfer ◽  
Articular Cartilage Defects

scholarly journals Effect of Vertical or Beveled Chondral Defect Creation on Rim Deformation and Contact

Cartilage ◽  
2018 ◽  
Vol 10 (2) ◽  
pp. 222-228 ◽  
Author(s):  
Adam B. Yanke ◽  
Megan L. Konopka ◽  
Davietta C. Butty ◽  
Maximilian A. Meyer ◽  
Eric J. Cotter ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Subchondral Bone ◽  
Rank Test ◽  
Cartilage Defects ◽  
Micro Computed Tomography ◽  
Bone Contact ◽  
Defect Center ◽  
Tibial Cartilage ◽  
Contact Width ◽  
Articular Cartilage Defects

Objective To determine biomechanical effects of knee cartilage defect perimeter morphology based on cartilage strain and opposing subchondral bone contact. Design Articular cartilage defects were created in 5 bovine femoral condyles: group 1, 45° inner bevel with 8-mm rim; group 2, vertical with 8-mm rim; and group 3, 45° outer bevel with 8-mm base. Samples were placed into a custom-machined micro–computed tomography tube and subjected to 800 N of axial loading. DICOM data were used to calculate cartilage thickness 4 and 6 mm from the center, distance between tibial cartilage surface and femoral subchondral bone, and contact width between tibial cartilage and subchondral bone. Strain 4 mm from the center and both absolute and change in distance (mm) to subchondral bone were compared between groups 1 and 2 using paired t tests. Strain at 6 mm and distance changed, loaded distance, and contact width (mm) were compared between groups using the Friedman test with post hoc analysis using Wilcoxon signed rank test. Results No significant differences in rim strain were noted between groups 1 and 2 at 4 mm ( P = 0.10) and between groups 1, 2, and 3 at 6 mm ( P = 0.247) from the defect center. The loaded distance was significantly different between groups 1 and 3 ( P = 0.013). No significant change in distance to the subchondral bone was found between groups ( P = 0.156). The difference in subchondral bone contact area approached but did not reach significance ( P = 0.074). Conclusion When debriding focal articular cartilage defects, establishment of an inner bevel decreases tissue deformation and contact with opposing subchondral bone.

scholarly journals Immunohistochemistry of types I and II collagen in undecalcified skeletal tissues.

1983 ◽  
Vol 31 (3) ◽  
pp. 417-425 ◽  
Author(s):  
W A Horton ◽  
C Dwyer ◽  
R Goering ◽  
D C Dean
Keyword(s):  
Articular Cartilage ◽  
Growth Plate ◽  
Subchondral Bone ◽  
Type I Collagen ◽  
Sodium Ethoxide ◽  
Cartilage Matrix ◽  
Antigenic Determinants ◽  
Immunoperoxidase Staining ◽  
Type I ◽  
Intense Staining

Types I and II collagen were demonstrated in semithin sections of undecalcified human endochondral growth plate, articular cartilage, and subchondral bone. The effects of several different methods for fixation, embedding, exposing of antigenic determinants, and immunoperoxidase staining were examined. Fixation in buffered formalin and paraformaldehyde-lysine-periodate solution gave more intense staining for collagens than fixation in paraformaldehyde-gluaraldehyde or Bouin's solution. Specimens embedded in Spurr epoxy resin yielded intense and uniform staining of areas known to contain the particular collagens after the resin had been removed by sodium ethoxide. The staining was enchanced following enzymatic digestion, especially with protease V (Sigma). Staining sensitivity and specificity were comparable with the indirect conjugate and double peroxidase-antiperoxidase (PAP) techniques; the PAP method was less sensitive. Embedment in methacrylate resins proved unsatisfactory because of exaggerated immunostaining of mineralized sites in comparison to unmineralized areas of the same tissues. In the growth plate specimens, type I collagen was identified in the matrices of bone, periosteum, perichondrium, and in the cytoplasm of hypertrophic and degenerative chondrocytes. Type II collagen was found uniformly throughout the cartilage matrix and in spicules of unresorbed cartilage matrix located in subchondral bone. A similar staining pattern was observed for the articular cartilage, except that type I collagen was not detected in chondrocytes.

Cross-linked type I and type II collagenous matrices for the repair of full-thickness articular cartilage defects—A study in rabbits

Biomaterials ◽  
2003 ◽  
Vol 24 (19) ◽  
pp. 3255-3263 ◽  
Author(s):  
Pieter Buma ◽  
Jeroen S. Pieper ◽  
Tony van Tienen ◽  
Job L.C. van Susante ◽  
Peter M. van der Kraan ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cartilage Defects ◽  
Type I ◽  
Type Ii ◽  
Full Thickness ◽  
Articular Cartilage Defects

Effects of TGF-β Overexpression via rAAV Gene Transfer on the Early Repair Processes in an Osteochondral Defect Model in Minipigs

2018 ◽  
Vol 46 (8) ◽  
pp. 1987-1996 ◽  
Author(s):  
Magali Cucchiarini ◽  
Ann-Kathrin Asen ◽  
Lars Goebel ◽  
Jagadeesh K. Venkatesan ◽  
Gertrud Schmitt ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Transforming Growth Factor ◽  
Biological Activities ◽  
Direct Gene Transfer ◽  
Type I ◽  
Lacz Gene ◽  
Early Repair ◽  
Repair Processes ◽  
Transfer Strategies

Background: Application of the chondrogenic transforming growth factor beta (TGF-β) is an attractive approach to enhance the intrinsic biological activities in damaged articular cartilage, especially when using direct gene transfer strategies based on the clinically relevant recombinant adeno-associated viral (rAAV) vectors. Purpose: To evaluate the ability of an rAAV–TGF-β construct to modulate the early repair processes in sites of focal cartilage injury in minipigs in vivo relative to control (reporter lacZ gene) vector treatment. Study Design: Controlled laboratory study. Methods: Direct administration of the candidate rAAV–human TGF-β (hTGF-β) vector was performed in osteochondral defects created in the knee joint of adult minipigs for macroscopic, histological, immunohistochemical, histomorphometric, and micro–computed tomography analyses after 4 weeks relative to control (rAAV- lacZ) gene transfer. Results: Successful overexpression of TGF-β via rAAV at this time point and in the conditions applied here triggered the cellular and metabolic activities within the lesions relative to lacZ gene transfer but, at the same time, led to a noticeable production of type I and X collagen without further buildup on the subchondral bone. Conclusion: Gene therapy via direct, local rAAV–hTGF-β injection stimulates the early reparative activities in focal cartilage lesions in vivo. Clinical Relevance: Local delivery of therapeutic (TGF-β) rAAV vectors in focal defects may provide new, off-the-shelf treatments for cartilage repair in patients in the near future.

Sign in / Sign up

Export Citation Format

Share Document