Viral Inactivation of Human Osteochondral Grafts with Methylene Blue and Light.

OBJECTIVE: Cartilage injury is one of the most common disorders of synovial joints. Fresh osteochondral allografts are becoming a standard treatment; however, they are supply constrained with a potential risk of disease transmission. There are no known virucidal processes available for osteochondral allografts and most methods presently available are detrimental to cartilage. Methylene blue light treatment has been shown to be successful in the literature for viral inactivation of fresh frozen plasma. The purpose of this study was to determine the capacity of methylene blue light treatment to inactivate a panel of clinically relevant viruses inoculated onto osteochondral allografts. DESIGN: Osteochondral grafts recovered from human cadaveric knees were inoculated with one of the following viruses: bovine viral diarrhea virus (BVDV), hepatitis A virus (HAV), human immunodeficiency virus type 1 (HIV-1), porcine parvovirus (PPV), and pseudorabies virus (PrV). The samples were processed through a methylene blue light treatment, which consisted of an initial soak in nonilluminated circulating methylene blue at ambient temperature, followed by light exposure with circulating methylene blue at cool temperatures. The final titer was compared with the recovery control for the viral log reduction. RESULTS: HIV-1, BVDV, and PrV were reduced to nondetectable levels while HAV and PPV were reduced by 3.1 and 5.6 logs, respectively. CONCLUSIONS: The methylene blue light treatment was effective in reducing (a) enveloped DNA and RNA viruses to nondetectable levels and (b) nonenveloped DNA and RNA viruses of inoculated human osteochondral grafts by 3.1 to 5.6 logs. This study demonstrates the first practical method for significantly reducing viral load in osteochondral implants.

Development of chondroitin sulfate encapsulated PLGA microsphere delivery systems with controllable multiple burst releases for treating osteoarthritis.

The purpose of the study was to design and develop unique drug delivery systems with controllable multiple burst releases of drugs for treating osteoarthritis. Chondroitin sulfate (CS) was encapsulated into four types of PLGA materials, that is, PLGA 50:50, PLGA 65:35, PLGA 75:25, and PLGA 85:15. The effects of microsphere size and various combinations of blend PLGA microspheres on CS release were investigated. The cytotoxicity of the CS-encapsulated microspheres was investigated according to the ISO 10993 guideline. Our study showed that the encapsulation efficiency of CS into PLGA 50:50 microspheres varied with the size of microspheres; however, the encapsulation efficiencies of CS into PLGA microspheres were independent of the types of PLGA materials. The size of PLGA microspheres was shown to affect the rate of CS release. With the increase of microsphere size from 75-150 µm to 300-355 µm, the initial CS release decreased. Further increase in microsphere size led to an increase in the initial CS release. In addition, combination of different types of PLGA microspheres was shown to be capable of achieving multiple burst CS releases. Moreover, the CS encapsulated PLGA microspheres were shown to be non-cytotoxic. This study proved the concept of multiple burst drug releases that were achieved by encapsulating CS into different types of PLGA microspheres and delivering CS from systems consisting of mixed types of PLGA microspheres, which may be applied to treat osteoarthritis by mimicking multiple intra-joint injection of therapeutic agents.

The bone marrow lesion in osteoarthritis.

Osteoarthritis (OA) is considered a multifactorial disease whose development and progression may include several structural abnormalities aside from cartilage destruction. Bone marrow lesions (BMLs) have been reported to be associated with OA pathology, and several studies have advocated its close connection to the severity of joint structural alterations and pain, the main OA clinical manifestation. Hence, BMLs may not only affect subchondral bone and its neuronal and vascular structures but also negatively influence the adjacent tissues. Here, we analyze the pathophysiology and natural history of OA-associated BMLs and their potential relevance to the radiographic progression and severity of the disease. The notion that BMLs may be a precursor to additional articular abnormalities, can be a potential risk factor for development of OA, and may serve as an additional diagnostic tool and a therapeutic target are further discussed.

Chondral Defect Repair with Particulated Juvenile Cartilage Allograft.

OBJECTIVE: This case study of 4 patients followed for at least 2 years was conducted to evaluate a cartilage repair procedure that involves transplanting particulated juvenile allograft cartilage. DESIGN: A multicenter, prospective, single-arm, 25-subject case study was designed to evaluate clinical outcomes such as IKDC and KOOS scores as well as the extent and quality of repair with MRI. In addition, there is an option for the transplants to be biopsied at various time points after implantation (up to 5 years). Currently, 25 patients with 1 or 2 chondral lesions on the femoral condyles and trochlea have been enrolled and treated in the prospective study. RESULTS: The first 4 patients have completed an evaluation at 24 months postoperative follow-up. Improvements in clinical outcomes over the preoperative baseline data have been observed. CONCLUSIONS: The present report describes, for the first time, clinical intermediate-term results of a novel cartilage repair procedure that involves transplanting particulated juvenile cartilage tissue allograft into prepared cartilage lesions of the femoral condyles and/or trochlea. Clinical outcome data of 4 patients who have reached the 24-month postimplantation milestone indicate early positive outcomes and suggest that this technique is capable of improving clinical symptoms. MRI data suggest that defect filling is possible and persists to at least 2 years. Continued clinical evaluation of this technique is needed with extended follow-up of all 25 patients in the series.

2-year postoperative evaluation of a patient with a symptomatic full-thickness patellar cartilage defect repaired with particulated juvenile cartilage tissue.

This case report describes the early results of a 36-year-old man who underwent repair of a symptomatic full-thickness patellar cartilage defect with transplanted particulated juvenile articular cartilage. At 2 years postoperatively, the patient has experienced substantial clinical improvement in both pain and function when evaluated with both International Knee Documentation Committee subjective evaluation and Knee Injury and Osteoarthritis Outcome Score outcome measures. Two-year postoperative magnetic resonance imaging demonstrates fill of the defect with repair tissue and near complete resolution of preoperative subchondral bone edema. To the best of the authors' knowledge, this case report is the first to report clinical results of this new technique at 2 years postoperatively.

Comparison of Achilles tendon repair techniques in a sheep model using a cross-linked acellular porcine dermal patch and platelet-rich plasma fibrin matrix for augmentation.

The primary goal of this study was to evaluate a cross-linked acellular porcine dermal patch (APD), as well as platelet-rich plasma fibrin matrix (PRPFM), for repair of acute Achilles tendon rupture in a sheep model. The 2 surgically transected tendon ends were reapproximated in groups 1 and 2, whereas a gap was left between the tendon ends in group 3. APD was used to reinforce the repair in group 2, and autologous PRPFM was used to fill the gap, which was also reinforced with APD, in group 3. All sheep were humanely euthanized at 24 weeks after the repair, and biomechanical and histological testing were performed. Tensile strength testing showed a statistically significant difference in elongation between the operated limb and the unoperated contralateral limb in groups 1 and 3, but not in group 2. All operated tendons appeared healed with no apparent fibrosis under light and polarized microscopy. In group 1, all surgical separation sites were identifiable, and healing occurred via increasing tendon thickness. In group 2, healing occurred with new tendon fibers across the separation, without increasing tendon thickness in 2 out of 6 animals. Group 3 showed complete bridging of the gap, with no change in tendon thickness in 2 out of 6 animals. In groups 2 and 3, peripheral integration of the APD to tendon fibers was observed. These findings support the use of APD, alone or with PRPFM, to augment Achilles tendon repair in a sheep model.

Wear, delamination, and fatigue resistance of melt-annealed highly crosslinked UHMWPE cruciate-retaining knee inserts under activities of daily living.

The wear, delamination, and fatigue resistance of artificially aged gamma irradiation-sterilized conventional polyethylene (CPE) and gas-plasma-sterilized melt-annealed highly crosslinked polyethylene tibial inserts (HXPE) were compared. Six CPE and 12 HXPE (six irradiated at 58 kGy and six at 72 kGy) left knee inserts were wear tested for 5.5 million cycles (Mc) under loads and motions that mimic activities of daily living, such as walking, chair rise, stair ascent, and deep squatting. Another six HXPE (72 kGy) and six CPE inserts were also tested under conditions that could produce severe delamination for 8 Mc. Ten other knees (five 72 kGy HXPE and five CPE) were subjected to posterior edge loading fatigue testing for 5 Mc. The HXPE inserts had an average wear rate reduction of about 80% relative to their CPE counterparts during all activities. All of the CPE inserts delaminated and fractured during high cycle deep squat (152 degrees flexion) motions, while all the HXPE remained intact. None of the HXPE inserts delaminated after 8 Mc, while all of the CPE inserts developed delamination damage within 1.5-5.8 Mc of delamination testing. All CPE inserts developed subsurface cracks and delamination within 2.8 Mc during posterior edge loading fatigue studies, while none of the HXPE inserts showed cracking or delamination after 5 Mc. These results show that aged HXPE has higher wear and fatigue resistance than aged CPE, and offers potential long-term advantages for young active patients with sustained activities of daily living.

In vivo outcomes of tissue-engineered osteochondral grafts.

Tissue-engineered osteochondral grafts have been synthesized from a variety of materials, with some success at repairing chondral defects in animal models. We hypothesized that in tissue-engineered osteochondral grafts synthesized by bonding mesenchymal stem cell-loaded hydrogels to a porous material, the choice of the porous scaffold would affect graft healing to host bone, and the quality of cell restoration at the hyaline cartilage surface. Bone marrow-derived allogeneic mesenchymal stem cells were suspended in hydrogels that were attached to cylinders of porous tantalum metal, allograft bone, or a bioactive glass. The tissue-engineered osteochondral grafts, thus created were implanted into experimental defects in rabbit knees. Subchondral bone restoration, defect fill, bone ingrowth-implant integration, and articular tissue quality were compared between the three subchondral materials at 6 and 12 weeks. Bioactive glass and porous tantalum were superior to bone allograft in integrating to adjacent host bone, regenerating hyaline-like tissue at the graft surface, and expressing type II collagen in the articular cartilage.

Fixation of tissue-engineered human neocartilage constructs with human fibrin in a caprine model.

The primary objective of this study was to determine the retention rate of neocartilage constructs in caprine full-thickness cartilage defects by fibrin sealant alone. Two defects, one each on the trochlea and the medial femoral condyle, were created in the stifle joint in 12 goats. Eight goats (16 defects) were treated with neocartilage constructs and 4 goats (8 defects) with fibrin glue alone. Postoperative activity was protected weight bearing for 6 weeks and then unrestricted for 18 weeks. At 24-week procurement, 4 neocartilage constructs were retained in 8 medial femoral defects and 4 in 8 trochlear defects. In gross comparison, the defects that retained the construct had a mean grade significantly higher than defects treated with fibrin glue alone. The mean histological score of defects with retained constructs was also higher than those treated with fibrin sealant alone. There was no appreciable immunologic reaction to the human neocartilage xenograft or human fibrin sealant.

The effect of entrapped bone particles on the surface morphology and wear of polyethylene.

Clinically retrieved highly cross-linked ultrahigh molecular weight polyethylene (HXPE) acetabular liners have demonstrated scratching, whereas conventional ultrahigh-molecular-weight polyethylene (UHMWPE) implants show a smoother surface early after implantation. In the present study, the potential of bone particles and soft tissues, rather than cement, to scratch the articular surface of HXPE and UHMWPE (gamma radiated) acetabular components was evaluated; multiple bone particles located at the articular surface for 3600 simulated walking cycles replicated the scratches observed on retrieved implants. By remelting, these scratches were confirmed to be due to plastic deformation of the polyethylene, not wear. Furthermore, it was shown using wear testing that these scratches did not affect the subsequent wear rate of HXPE or conventional UHMWPE. Wear rates of scratched conventional and cross-linked polyethylene were not significantly different from unscratched conventional and cross-linked polyethylene, respectively.

The interleukin 1beta pathway in the pathogenesis of osteoarthritis.

Osteoarthritis (OA) is a major disabling disease and is ranked as a major cause of chronic pain in adults. The pathology of the illness is characterized by a loss of articular cartilage leading to narrowing of joint space, increased joint friction, potential structural remodeling, persistent pain, and functional impairment. The proinflammatory cytokine interleukin 1beta (IL-1beta) has several chemical and bioactive characteristics allowing this catabolic protein to be involved in initiation and progression of OA. We review the current understanding of the pathogenesis of OA, and how upregulation of IL-1beta initiates a cascade of intracellular events that can culminate in activation of proteinases, creation of a pro-destructive articular milieu, suppression of anabolic pathways, and a decrease in the synthesis of cartilage extracellular matrix. Therapeutic approaches to block the action of IL-1beta and overcome its signal transduction to curtail disease progression are discussed.

Characterization of a highly cross-linked ultrahigh molecular-weight polyethylene in clinical use in total hip arthroplasty.

This article reports on a commercially available extensively cross-linked ultrahigh molecular-weight polyethylene (HXPE) produced by subjecting molded GUR 1050 ultrahigh molecular-weight polyethylene (UHMWPE) to 100 +/- 10 kGy of electron beam radiation followed by melt annealing and sterilization by gas plasma. When compared to contemporary conventional molded GUR 1050 UHMWPE sterilized by 37 kGy of gamma radiation, the HXPE material has enhanced wear properties, has no detectable free radicals, and is resistant to oxidation and oxidative-related material property changes. The relative wear improvement of the HXPE is maintained in the presence of bone cement or alumina particles. The HXPE produced greater than 90% fewer wear particles in all size ranges and statistically significantly (P < .0001) smaller average-size particles than did the conventional UHMWPE.

Evaluation of a cross-linked acellular porcine dermal patch for rotator cuff repair augmentation in an ovine model.

In this study we evaluated 2 commercially available rotator cuff repair augmentation patches in an in vivo sheep model using mechanical testing and histologic techniques. Bilateral infraspinatus tears were created and repaired in 2 groups of 8 adult ewes. Each group (killed at 9 or 24 weeks) included 5 repaired with suture alone, 6 repaired and augmented with a cross-linked acellular porcine dermal (PD) patch (Zimmer Collagen Repair Patch), and 5 repaired and augmented with a porcine small intestine submucosa (SIS) patch (Restore Orthobiologic Soft Tissue Implant; DePuy Orthopaedics). At 3 weeks, sheep with suture repair and an SIS patch had significant elevation of plasma fibrinogen levels (P < .05) whereas sheep with suture repair and a PD patch elicited no elevation in plasma fibrinogen levels. At 9 weeks, the mean failure load was 201 +/- 60 lb for suture repairs, 182 +/- 63 lb for PD repairs, and 137 +/- 16 lb for SIS repairs. Within any individual sheep, the shoulder undergoing PD repair always had a higher failure load than the contralateral suture or shoulder undergoing SIS repair. At 9 weeks, macrophages were seen on all PD surfaces whereas most of the SIS materials were resorbed. At 24 weeks, failure loads were identical between groups. Macrophages had disappeared from the PD groups, and integration of the PD patch into the surrounding tissue with vascular and fibroblastic invasion was seen. For the SIS group, diverse tissue types (including ectopic bone) were seen.

Hydrostatic pressure modulates proteoglycan metabolism in chondrocytes seeded in agarose.

OBJECTIVE: To investigate the effect of isolated hydrostatic pressure on proteoglycan metabolism in chondrocytes. METHODS: Bovine articular chondrocytes cultured in agarose gels were subjected to 5 MPa hydrostatic pressure for 4 hours in either a static or a pulsatile (1 Hz) mode, and changes in glycosaminoglycan (GAG) synthesis, hydrodynamic size, and aggregation properties of proteoglycans and aggrecan messenger RNA (mRNA) levels were determined. RESULTS: The application of 5 MPa static pressure caused a significant increase in GAG synthesis of 11% (P < 0.05). Column chromatography showed that this increase in GAG synthesis was associated with large proteoglycans. In addition, semiquantitative reverse transcriptase-polymerase chain reaction showed a 4-fold increase in levels of aggrecan mRNA (P < 0.01). CONCLUSION: Hydrostatic pressure in isolation, which does not cause cell deformation, can affect proteoglycan metabolism in chondrocytes cultured in agarose gels, indicating an important role of hydrostatic pressure in the regulation of extracellular matrix turnover in articular cartilage.