Bonding and fusion of meniscus fibrocartilage using a novel chondroitin sulfate bone marrow tissue adhesive.

The weak intrinsic meniscus healing response and technical challenges associated with meniscus repair contribute to a high rate of repair failures and meniscectomies. Given this limited healing response, the development of biologically active adjuncts to meniscal repair may hold the key to improving meniscal repair success rates. This study demonstrates the development of a bone marrow (BM) adhesive that binds, stabilizes, and stimulates fusion at the interface of meniscus tissues. Hydrogels containing several chondroitin sulfate (CS) adhesive levels (30, 50, and 70 mg/mL) and BM levels (30%, 50%, and 70%) were formed to investigate the effects of these components on hydrogel mechanics, bovine meniscal fibrochondrocyte viability, proliferation, matrix production, and migration ability in vitro. The BM content positively and significantly affected fibrochondrocyte viability, proliferation, and migration, while the CS content positively and significantly affected adhesive strength (ranged from 60±17 kPa to 335±88 kPa) and matrix production. Selected material formulations were translated to a subcutaneous model of meniscal fusion using adhered bovine meniscus explants implanted in athymic rats and evaluated over a 3-month time course. Fusion of adhered meniscus occurred in only the material containing the highest BM content. The technology can serve to mechanically stabilize the tissue repair interface and stimulate tissue regeneration across the injury site.

An adhesive bone marrow scaffold and bone morphogenetic-2 protein carrier for cartilage tissue engineering.

A chondroitin sulfate-bone marrow (CS-BM) adhesive hydrogel was used to localize rhBMP-2 to enhance articular cartilage tissue formation. Chondrocyte pellet culture revealed that 0.1 and 1 µg/mL of rhBMP-2 enhanced sulfated-GAG content. rhBMP-2 localization within the hydrogels was investigated, and it was found that BM, CS-NHS, and rhBMP-2 levels and time affected rhBMP-2 retention. Retention was modulated from 82 to 99% over a 3-week period for the material formulations investigated. To evaluate carrier efficacy, rhBMP-2 and bovine articular chondrocytes were encapsulated within CS-BM, and biochemical evaluation revealed significant increases in total collagen production with rhBMP-2. Histological analysis revealed more robust tissue formation and greater type-II collagen production with encapsulated rhBMP-2. Subsequently, a subcutaneous culture of hydrogels revealed increased total collagen, type-II to type-I collagen ratio, and sulfated GAG in samples carrying rhBMP-2. These findings indicate the development of a multifunctional system capable of localizing rhBMP-2 to enhance repair tissue quality.

An orthopedic tissue adhesive for targeted delivery of intraoperative biologics.

Tissue adhesives can bind together damaged tissues and serve as tools to deliver and localize therapeutics to facilitate regeneration. One emerging therapeutic trend in orthopedics is the use of intraoperative biologics (IOB), such as bone marrow (BM) and platelet-rich plasma (PRP), to stimulate healing. Here, we introduce the application of the biomaterial chondroitin sulfate succinimidyl succinate (CS-NHS) to deliver IOB in a hydrogel adhesive. We demonstrate the biomaterial's ability to bind various tissue types and its cellular biocompatibility with encapsulated human mesenchymal stem cells (hMSCs). Further, we examine in detail the CS-NHS adhesive combined with BM aspirate for use in bone applications. hMSCs were encapsulated in CS-BM and cultured for 5 weeks in osteogenic medium. Quantitative RT-PCR demonstrated osteogenesis via upregulation of the osteogenic transcription factor Runx2 and bone markers alkaline phosphatase and osteocalcin. Significant deposition of calcium and osteocalcin was detected using biochemical, histological, and immunohistochemical techniques. Shear testing demonstrated that the CS-BM adhesive exhibited an adhesive strength approximately an order of magnitude stronger than fibrin glue and approaching that of a cyanoacrylate adhesive. These results indicate that CS-NHS is a promising delivery tool for IOB in orthopedic applications requiring a strong, degradable, and biocompatible adhesive that supports bone growth.

Lubricin distribution in the human intervertebral disc.

BACKGROUND: Previous studies have identified lubricin (also known as superficial zone protein) as a lubricating glycoprotein present in several musculoskeletal tissues including articular cartilage, meniscus, and tendon. In this immunohistochemical study, we determined the presence and distribution of lubricin in the cells, extracellular matrix, and tissue surfaces of human nucleus pulposus and anulus fibrosus tissues. METHODS: Twenty-eight human intervertebral discs were resected at autopsy from fourteen cadavers. Disc specimens were fixed in formalin, processed, and paraffin-embedded prior to sectioning. Tissue sections were immunohistochemically stained for lubricin, the extent of extracellular matrix staining was evaluated semiquantitatively, and cellular staining was assessed quantitatively with use of a survey method. RESULTS: Lubricin staining was evident in the extracellular matrix and at select surfaces of the nucleus pulposus and anulus fibrosus tissues. The extent of lubricin staining of the extracellular matrix was contingent on the disc region (nucleus pulposus, inner anulus fibrosus, or outer anulus fibrosus), with the greatest extent of matrix staining found in the nucleus pulposus, but it was not contingent on the Thompson grade. A subset of disc cells within the nucleus, inner anulus, and outer anulus also stained positively for lubricin, suggesting intrinsic cell synthesis of the glycoprotein. The disc region significantly affected the percentage of lubricin-staining cells, with the greatest percentage of cells staining for lubricin (nearly 10%) found in the nucleus pulposus. The percentage of cells staining for lubricin correlated with the extent of extracellular matrix staining for lubricin. CONCLUSIONS: The results of this study confirm the presence of lubricin in the human intervertebral disc and demonstrate a unique distribution compared with that in the goat. The presence of lubricin in asymptomatic discs provides a foundation for future research regarding the role of lubricin in pathological disc conditions.