Autologous collagen induced chondrogenesis (ACIC: Shetty–Kim technique) – A matrix based acellular single stage arthroscopic cartilage repair technique

The defects of articular cartilage in the knee joint are a common degenerative disease and currently there are several established techniques to treat this problem, each with their own advantages and shortcomings. Autologous chondrocyte implantation is the current gold standard but the technique is expensive, time-consuming and most versions require two stage procedures and an arthrotomy. Autologous collagen induced chondrogenesis (ACIC) is a single-stage arthroscopic procedure and we developed. This method uses microfracture technique with atelocollagen mixed with fibrin gel to treat articular cartilage defects. We introduce this ACIC techniques and its scientific background.     

Modified osteochondral autograft implantation for full- thickness articular cartilage lesions

Full-thickness articular cartilage defects have been difficult to treat in patients with nonarthritic knees. A procedure is described to treat articular cartilage full-thickness lesions. Graft sites are chosen after appropriate treatment of the base of a grade IV lesion. Articular cartilage and bone are replaced into the graft site, promoting mesenchymal stem cell growth and cartilaginous coverage of the defect. Pathology, postoperative protocol, and some postoperative arthroscopic illustrations are included. This technique is simple and is associated with minimal donor-site morbidity.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 19, No 3 (March), 2003: pp 318–320     

Editorial Commentary: Microfracture Remains a Foundational Tool for Cartilage Restoration During Hip-Preservation Procedures: Defying the Odds

A comprehensive approach to arthroscopic hip preservation in patients whose pathology includes cartilage defects may include microfracture; microfracture has been shown to have long-lasting positive effects in most patients treated for femoroacetabular impingement plus full-thickness chondral pathology. Although modern cartilage treatment alternatives such as autologous chondrocyte implantation, autologous matrix-induced chondrogenesis scaffolds, allograft or autograft particulate cartilage graft, and others have been described for the treatment of high-degree cartilage acetabular lesions, microfracture remains a foundational tool in cartilage restoration procedures. That said, when determining outcome, comorbidity must be considered, and, moreover, it is difficult to determine whether outcomes are only attributable to the microfracture versus concomitant procedures or changes in postoperative activity of operated patients.     

Marrow stimulation techniques

Due to the very low intrinsic activity of human adult cartilage, healing of chondral and osteochondral defects in patients cannot be expected. In treating symptomatic cartilage damage, marrow stimulation methods belong to the most frequently used methods, along with autologous chondrocyte transplantation (ACT) and mosaicplasty. These arthroscopic procedures are generally easy and the marrow stimulation treatment costs relatively little. In recent years, Pridie drilling has been increasingly replaced by the microfracture technique. This modification relies on the same biological principles of promoting resurfacing with the formation of fibro-cartilaginous repair tissue. For the treatment of smaller cartilage defects (<2.5 cm(2)), microfracture still remains the first choice for treatment. The clinical results after microfracture in the knee are age dependent. Younger and active patients (<40 years) with smaller isolated traumatic lesions on the femoral condyles have the best long-term results. The deterioration of the clinical results begins after 18 months and is significantly more pronounced in older patients with defects on the patella-femoral joint and tibia. The inferior quality of the repair tissue, partially incomplete defect filling and new bone formation in the defect area seem to be limitations of these methods. The AMIC (autologous matrix induced chondrogenesis) technique was developed to enable treatment of larger defects by the application of a collagen Type III/I membrane (Geistlich Pharma, Wolhusen, Switzerland), in particular when cell-engaged procedures such as ACT cannot be used for financial reasons or because it is not indicated. AMIC seems to be particularly suitable for treating damaged retropatellar cartilage, which is an advantage because these defects can be hard to treat with standard microfracturing alone. The results of the ongoing studies are awaited to establish whether better results with this technology are achievable in the long term.     

Second-look arthroscopic observations after radiofrequency treatment of partial thickness articular cartilage defects in human knees: report of four cases

Partial thickness articular cartilage defects in the knee are commonly encountered clinical problems. Recently, use of radiofrequency-based devices for performing arthroscopic chondroplasty has gained popularity. However, published experimental studies using different methods for evaluating the histologic effects of radiofrequency-chondroplasty on surrounding cartilage offer contradictory results. To date, few clinical findings after radiofrequency-based chondroplasty have been reported. We present four patients where follow-up arthroscopy documented partial thickness articular defects treated previously with radiofrequency-based chondroplasty to be completely filled with stable repair tissue. No attempt was made to stimulate cartilage regeneration (ie, abrasion or microfracture) in any of these cases.     

Formation of cartilage repair tissue in articular cartilage defects pretreated with microfracture and covered with cell‐free polymer‐based implants

The aim of our study was to evaluate the mid‐term outcome of a cell‐free polymer‐based cartilage repair approach in a sheep cartilage defect model in comparison to microfracture treatment. Cell‐free, freeze‐dried implants (chondrotissue®) made of a poly‐glycolic acid (PGA) scaffold and hyaluronan were immersed in autologous serum and used for covering microfractured full‐thickness articular cartilage defects of the sheep (n = 4). Defects treated with microfracture only served as controls (n = 4). Six months after implantation, cartilage implants and controls were analyzed by immunohistochemical staining of type II collagen, histological staining of proteoglycans, and histological scoring. Histological analysis showed the formation of a cartilaginous repair tissue rich in proteoglycans. Histological scoring documented significant improvement of repair tissue formation when the defects were covered with the cell‐free implant, compared to controls treated with microfracture. Immunohistochemistry showed that the cell‐free implant induced cartilaginous repair tissue and type II collagen. Controls treated with microfracture showed marginal formation of a mixed‐type repair tissue consisting of cartilaginous tissue and fibro‐cartilage. Covering of microfractured defects with the cell‐free polymer‐based cartilage implant is suggested to be a promising treatment option for cartilage defects and improves the regeneration of articular cartilage. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1353–1360, 2009     

Arthroskopische M?glichkeiten biorekonstruktiver Verfahren bei Knorpelsch?den der Schulter

Chondral or osteochondral lesions of the shoulder may lead to premature osteoarthritis of the glenohumeral joint as regeneration of damaged articular cartilage is lacking. Rising health awareness, increasingly active populations and improvements in medical techniques have increased the application of cartilage regenerative minimally invasive approaches for glenohumeral joint preservation or delayed prosthetic replacement. In contrast to the conclusive and mostly convincing mid-term results of cartilage regenerative techniques known for the knee, clinical results of innovative therapeutic approaches with glenohumeral cartilage defects are more or less absent. Current techniques include procedures for mesenchymal stem cell recruitment, such as microfracturing, (autologous) osteochondral transplantation, (matrix-associated) autologous chondrocyte transplantation and biological resurfacing, addressing focal chondral defects up to massive structural osteochondral defects. With increasing arthroscopic applicability, they evolve to important tools in the armamentarium of the shoulder surgeon. Future clinical data will determine evidence-based applicability, enabling standardized treatment selection.     

Combined effects of insulin-like growth factor-1 and transforming growth factor-beta1 on periosteal mesenchymal cells during chondrogenesis in vitro

OBJECTIVE: Periosteum contains undifferentiated mesenchymal stem cells that have both chondrogenic and osteogenic potential, and has been used to repair articular cartilage defects. During this process, the role of growth factors that stimulate the periosteal mesenchymal cells toward chondrogenesis to regenerate articular cartilage and maintain its phenotype is not yet fully understood. In this study, we examined the effects of insulin-like growth factor-1 (IGF-1) and transforming growth factor-beta1 (TGF-beta1), alone and in combination, on periosteal chondrogenesis using an in vitro organ culture model. METHODS: Periosteal explants from the medial proximal tibia of 2-month-old rabbits were cultured in agarose under serum free conditions for up to 6 weeks. After culture the explants were weighed, assayed for cartilage production via Safranin O staining and histomorphometry, assessed for proliferation via proliferative cell nuclear antigen (PCNA) immunostaining, and assessed for type II collagen mRNA expression via in situ hybridization. RESULTS: IGF-1 significantly increased chondrogenesis in a dose-dependent manner when administered continuously throughout the culture period. Continuous IGF-1, in combination with TGF-beta1 for the first 2 days, further enhanced overall total cartilage growth. Immunohistochemistry for PCNA revealed that combining IGF-1 with TGF-beta1 gave the strongest proliferative stimulus early during chondrogenesis. In situ hybridization for type II collagen showed that continuous IGF-1 maintained type II collagen mRNA expression throughout the cambium layer from 2 to 6 weeks. CONCLUSION: The results of this study demonstrate that IGF-1 and TGF-beta1 can act in combination to regulate proliferation and differentiation of periosteal mesenchymal cells during chondrogenesis.     

Active proliferation of mesenchymal cells prior to the chondrogenic repair response in rabbit full-thickness defects of articular cartilage

OBJECTIVES: In full-thickness articular defects, fibroblast growth factor-2 (FGF-2) participates in the chondrogenic repair response which occurs in a defect-size dependent manner. Here we demonstrate that FGF-2 plays a critical role in the proliferation of pre-chondrogenic mesenchymal cells during chondrogenic induction. METHODS: Three-millimeter- or 5-mm-diameter cylindrical defects were created in the femoral trochlea of the rabbit knee. The defects received sterile saline or FGF-2 (50 pg/h) via an osmotic pump for the initial 2 weeks. We assessed the proliferative capacity of undifferentiated mesenchymal cells in the reparative tissue with the anti-proliferating cell nuclear antigen (PCNA) monoclonal antibody. Using a total of 180 rabbits, we performed three sets of experiments. RESULTS: In the 3-mm-diameter defects, undifferentiated mesenchymal cells spontaneously initiated chondrogenic differentiation within 2 weeks, resulting in the regeneration of surfacing articular cartilage concomitantly with the repair of subchondral bone. No evidence of chondrogenesis was seen in the 5-mm-diameter defects, whereas application of FGF-2 promoted successful regeneration of articular cartilage. In the 3-mm-diameter defects and in the FGF-2-treated 5-mm defects, PCNA immunoreactivity was widely detected in undifferentiated cells in the reparative tissue at 1 and 2 weeks after creation of the defects. In contrast, in the 5-mm-diameter defects without FGF-2 treatment, the PCNA-positive cells were found at a significantly lower incidence. CONCLUSIONS: Active expansion of undifferentiated cell population mediated by FGF-2 is required to initiate and support a chondrogenic repair response in full-thickness defects of articular cartilage. Endogenous FGF-2 could not meet the requirements of growth signaling in the center of larger sized defects.     

Analyses of early events during chondrogenic repair in rat full-thickness articular cartilage defects

In this study we investigated the cellular events that occur during the onset of chondrogenic differentiation during the repair of full-thickness defects of articular cartilage. The V-shaped full-thickness cartilage defects (width 0.7 or 1.5 mm; depth 0.8 mm; length 4 mm) were created in the femoral patellar groove of rats using a custom-built twin-blade device. The time course of the repair response in these cartilage defects was examined using a semi-quantitative histological grading scale. Cartilaginous repair responses failed to occur in the larger 1.5 mm defects, which was covered only by fibrous scar tissue. In contrast, hyaline-like articular cartilage was regenerated concomitantly with the repair of the subchondral bone by 4 weeks in smaller 0.7 mm width defects. Cells in the reparative regions were then characterized by immunohistochemistry and in situ hybridization. Undifferentiated mesenchymal cells migrate into the defects and fill the cavities within 4 days of their creation. The expression of PCNA, N-cadherin, and PTH/PTHrP receptors was induced in cells at the center of the defects, where type II collagen-positive polygonal-shaped cells also begin to appear at day 7. Marrow-derived mesenchymal cells acquire higher levels of proliferative activity in induced cartilage cavities after their initial migration and filling of the smaller 0.7 mm defects. During the regenerative repair of articular cartilage in the rat, there is a distinctive step that appears to be analogous to the precartilaginous condensation that is pivotal during chondrogenesis in development.     

Effects of harvest and selected cartilage repair procedures on the physical and biochemical properties of articular cartilage in the canine knee.

This study utilizes a canine model to quantify changes in articular cartilage 15-18 weeks after a knee joint is subjected to surgical treatment of isolated chondral defects. Clinical and experimental treatment of articular cartilage defects may include implantation of matrix materials or cells, or both. Three cartilage repair methods were evaluated: microfracture, microfracture and implantation of a type-II collagen matrix, and implantation of an autologous chondrocyte-seeded collagen matrix. The properties of articular cartilage in other knee joints subjected to harvest of articular cartilage from the trochlear ridge (to obtain cells for the cell-seeded procedure) were also evaluated. Physical properties (thickness, equilibrium compressive modulus, dynamic compressive stiffness, and streaming potential) and biochemical composition (hydration, glycosaminoglycan content, and DNA content) of the cartilage from sites distant to the surgical treatment were compared with values measured for site-matched controls in untreated knee joints. No significant differences were seen in joints subjected to any of the three cartilage repair procedures. However, a number of changes were induced by the harvest operation. The largest changes (displaying up to 3-fold increases) were seen in dynamic stiffness and streaming potential of patellar groove cartilage from joints subjected to the harvest procedure. Whether the changes reported will lead to osteoarthritic degeneration is unknown, but this study provides evidence that the harvest procedure associated with autologous cell transplantation for treatment of chondral defects may result in changes in the articular cartilage in the joint.     

Knochenmarkstimulierende Techniken zur Knorpeldefektbehandlung: gestern, heute und morgen

Various surgical techniques are now available for the treatment of local chondral defects, but there is still no general agreement on the type of treatments indicated. The treatment options are divided basically into procedures with and without chondrocyte transplantation, the latter being subdivided into such procedures as marrow stimulation and transplantation. The most common surgical technique used in the treatment of local chondral defects is microfracturing, in which subchondral bone spaces are opened so that a blood clot fills the defect. In recent years this technique has been improved by the addition of autologous matrix-induced chondrogenesis (AMIC). The application of a matrix for better containment and of fibrin glue has made it possible to treat larger chondral lesions. Further advantages over autologous chondrocyte implantation are the lower cost and the single-step immediate treatment of the cartilage defects, since an additional cartilage biopsy for cell culture is not needed. Further development of matrices and growth factors will continue in the future. The present article gives an overview of the development of microfracture, the indications for it, and the current therapeutic alternatives in cartilage repair.     

Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees

OBJECTIVE: There is no widely accepted method to repair articular cartilage defects. Bone marrow mesenchymal cells have the potential to differentiate into bone, cartilage, fat and muscle. Bone marrow mesenchymal cell transplantation is easy to use clinically because cells can be easily obtained and can be multiplied without losing their capacity of differentiation. The objective of this study was to apply these cell transplantations to repair human articular cartilage defects in osteoarthritic knee joints. DESIGN: Twenty-four knees of 24 patients with knee osteoarthritis (OA) who underwent a high tibial osteotomy comprised the study group. Adherent cells in bone marrow aspirates were culture expanded, embedded in collagen gel, transplanted into the articular cartilage defect in the medial femoral condyle and covered with autologous periosteum at the time of 12 high tibial osteotomies. The other 12 subjects served as cell-free controls. RESULTS: In the cell-transplanted group, as early as 6.3 weeks after transplantation the defects were covered with white to pink soft tissue, in which metachromasia was partially observed. Forty-two weeks after transplantation, the defects were covered with white soft tissue, in which metachromasia was observed in almost all areas of the sampled tissue and hyaline cartilage-like tissue was partially observed. Although the clinical improvement was not significantly different, the arthroscopic and histological grading score was better in the cell-transplanted group than in the cell-free control group. CONCLUSIONS: This procedure highlights the availability of autologous culture expanded bone marrow mesenchymal cell transplantation for the repair of articular cartilage defects in humans.     

Microfracture and bone morphogenetic protein 7 (BMP-7) synergistically stimulate articular cartilage repair

OBJECTIVE: Microfracture is used to treat articular cartilage injuries, but leads to the formation of fibrocartilage rather than native hyaline articular cartilage. Since bone morphogenetic protein 7 (BMP-7) induces cartilage differentiation, we hypothesized that the addition of the morphogen would improve the repair tissue generated by microfracture. We determined the effects of these two treatments alone and in combination on the quality and quantity of repair tissue formed in a model of full-thickness articular cartilage injury in adolescent rabbits. DESIGN: Full-thickness defects were made in the articular cartilage of the patellar grooves of forty, 15-week-old rabbits. Eight animals were then assigned to (1) no further treatment (control), (2) microfracture, (3) BMP-7, (4) microfracture with BMP-7 in a collagen sponge (combination treatment), and (5) microfracture with a collagen sponge. Animals were sacrificed after 24 weeks at 39 weeks of age. The extent of healing was quantitated by determining the thickness and the surface area of the repair tissue. The quality of the repair tissue was determined by grading specimens using the International Cartilage Repair Society Visual Histological Assessment Scale. RESULTS: Compared to controls, BMP-7 alone increased the amount of repair tissue without affecting the quality of repair tissue. Microfracture improved both the quantity and surface smoothness of repair tissue. Compared to either single treatment, the combination of microfracture and BMP-7 increased both the quality and quantity of repair tissue. CONCLUSIONS: Microfracture and BMP-7 act synergistically to stimulate cartilage repair, leading to larger amounts of repair tissue that more closely resembles native hyaline articular cartilage.     

Editorial Commentary: Second-Generation Microfracture—We Are Only As Strong As Our Weakest Link

Injuries to the articular cartilage of the knee are increasingly common, especially in athletes. The operative management of these focal chondral lesions continues to be a regenerative challenge. The microfracture (MFx) procedure has become a first-line arthroscopic treatment method for small, symptomatic chondral lesions, and it frequently serves as the standard technique against which other cartilage repair procedures are compared. Over time, outcome studies have defined the weaknesses and limitations of first-generation MFx. The second iteration of MFx seeks to optimize regeneration using the trilogy of cells, scaffolds, and growth factors. As surgeons, we are only as strong as our weakest link.     

髌下脂肪垫来源间充质干细胞软骨分化的研究进展

间充质干细胞(MSCs)为重点的组织工程学研究已成为软骨损伤修复与再生的研究热点,髌下脂肪垫(IPFP)是脂肪间充质干细胞(ASCs)的新型优良组织来源,其取材方便,供区损伤小。且IPFP-ASCs体外增殖快,成软骨分化能力较强。低氧,转化生长因子β1、β3,骨形态发生蛋白7等可以促进其分化。体内试验表明其可以有效改善软骨损伤患者症状,提高关节功能。虽然目前仍存在生物学和技术上的困难需克服,但IPFP-ASCs有望成为修复包括骨关节炎在内的关节软骨损伤的良好策略。     

The microfracture technique to treat full thickness articular cartilage defects of the knee

Full thickness defects of the articular cartilage rarely heal spontaneously. While some patients do not develop clinically significant problems from chondral defects, most eventually develop degenerative changes associated with the cartilage damage over time. Techniques to treat chondral defects include abrasion, drilling, tissue autografts, allografts, and cell transplantation. The senior author has developed a procedure referred to as the "microfracture." This technique enhances chondral resurfacing by providing a suitable environment for tissue regeneration and by taking advantage of the body's own healing potential. This technique has now been used in more than 1400 patients. Specially designed awls are used to make multiple perforations, or "microfractures", into the subchondral bone plate. The perforations are made as close together as necessary, but not so close that one breaks into another. Consequently, the microfracture holes are approximately three to four millimeters apart (or 3 to 4 holes per square centimeter). Importantly, the integrity of the subchondral bone plate is maintained. The released marrow elements form a "super clot" which provides an enriched environment for tissue regeneration. Follow up with long term results of more than 8 years have been positive and very encouraging.     

Osteochondral grafting of knee joint using mosaicplasty

Focal cartilage defects of articular surface-traumatic and degenerative are difficult to treat, thus a variety of surgical techniques have been developed and reported for treatment of such defects. Procedures such as Priddies perforations, microfracture, abrasion chondroplasty have shown long-term results which are often less than adequate. One of the reasons is that all these techniques lead to the formation of fibrocartilage which has inferior mechanical properties as compared to the native hyaline cartilage. Mosaicplasty is a procedure which aims at replacing the lost articular cartilage with hyaline cartilage including underlying bone support, thus providing adequate stability to the cartilage and better cartilage/bone integration. A young man underwent this procedure for recalcitrant knee pain at our institution. At 2 years follow-up, his knee pain has significantly improved. We hereby present medium term results (2 years) of this first case report in local literature.     

Chondrogenic differentiation of human subchondral progenitor cells is affected by synovial fluid from donors with osteoarthritis or rheumatoid arthritis

Background

Microfracture is a first-line treatment option for cartilage repair. In microfracture, subchondral mesenchymal cortico-spongious progenitor cells (CSP) enter the defect and form cartilage repair tissue. The aim of our study was to investigate the effects of joint disease conditions on the in vitro chondrogenesis of human CSP.

Methods

CSP were harvested from the subchondral bone marrow. CSP characterization was performed by analysis of cell surface antigen pattern and by assessing the chondrogenic, osteogenic and adipogenic differentiation potential, histologically. To assess the effect of synovial fluid (SF) on chondrogenesis of CSP, micro-masses were stimulated with SF from healthy (ND), osteoarthritis (OA) and rheumatoid arthritis donors (RA) without transforming growth factor beta 3.

Results

CSP showed the typical cell surface antigen pattern known from mesenchymal stem cells and were capable of osteogenic, adipogenic and chondrogenic differentiation. In micro-masses stimulated with SF, histological staining as well as gene expression analysis of typical chondrogenic marker genes showed that SF from ND and OA induced the chondrogenic marker genes aggrecan, types II and IX collagen, cartilage oligomeric matrix protein (COMP) and link protein, compared to controls not treated with SF. In contrast, the supplementation with SF from RA donors decreased the expression of aggrecan, type II collagen, COMP and link protein, compared to CSP treated with SF from ND or OA.

Conclusion

These results suggest that in RA, SF may impair cartilage repair by subchondral mesenchymal progenitor cells in microfracture, while in OA, SF may has no negative, but a delaying effect on the cartilage matrix formation.