A Versatile, Secure, and Sustainable All-in-One Biobank-Registry Data Solution: The A3BC REDCap Model.

Introduction: A key element in the big data revolution is large-scale biobanking and the associated development of high-quality data collections and supporting informatics solutions. As such, in establishing the Australian Arthritis and Autoimmune Biobank Collaborative (A3BC), we sought to establish a low-cost, nation-scale data management system capable of managing a multisite biobank registry with complex longitudinal sample and data requirements. Materials and Methods: We assessed several international commercial and nonprofit software platforms using standardized system requirement criteria and follow-up interviews. Vendor compliance scoring was prioritized to meet our project-critical requirements. Consumer/end-user codesign was integral to refining our system requirements for optimized adoption. Customization of the selected software solution was performed to optimize field auto-population between participant timepoints and forms, using modules that are transferable and that do not impact core code. Institutional and independent testing was used to ensure data security. Results: We selected the widely used research web application Research Electronic Data Capture (REDCap), which is "free" (under nonprofit license agreement terms), highly configurable, and customizable to a variety of biobank and registry needs and can be developed/maintained by biobank users with modest IT skill, time, and cost. We created a secure, comprehensive participant-centric biobank-registry database that includes: (1) best practice data security measures (incl. multisite access login using institutional user credentials), (2) permission-to-contact and dynamic itemized electronic consent, (3) a complete chain of custody from consent to longitudinal biospecimen data collection to publication, (4) complex longitudinal patient-reported surveys, (5) integration of record-level extracted/linked participant data, (6) significant form auto-population for streamlined data capture, and (7) native dashboards for operational visualizations. Conclusion: We recommend the versatile, reusable, and sustainable informatics model we have developed in REDCap for prospective chronic disease biobanks or registry biobanks (of local to national complexity) supporting holistic research into disease prediction, precision medicine, and prevention strategies.

In vitro evaluation of natural marine sponge collagen as a scaffold for bone tissue engineering.

The selection of a suitable scaffold matrix is critical for cell-based bone tissue engineering. This study aimed to identify and characterize natural marine sponges as potential bioscaffolds for osteogenesis. Callyspongiidae marine sponge samples were collected from the Fremantle coast of Western Australia. The sponge structure was assessed using scanning electron microscopy (SEM) and Hematoxylin and eosin. Mouse primary osteoblasts were seeded onto the sponge scaffold and immunostained with F-actin to assess cell attachment and aggregation. Alkaline phosphatase expression, von Kossa staining and real-time PCR were performed to examine the osteogenic potential of sponge samples. SEM revealed that the sponge skeleton possessed a collagenous fibrous network consisting of interconnecting channels and a porous structure that support cellular adhesion, aggregation and growth. The average pore size of the sponge skeleton was measured 100 to 300 µm in diameter. F-actin staining demonstrated that osteoblasts were able to anchor onto the surface of collagen fibres. Alkaline phosphatase expression, a marker of early osteoblast differentiation, was evident at 7 days although expression decreased steadily with long term culture. Using von Kossa staining, mineralisation nodules were evident after 21 days. Gene expression of osteoblast markers, osteocalcin and osteopontin, was also observed at 7, 14 and 21 days of culture. Together, these results suggest that the natural marine sponge is promising as a new scaffold for use in bone tissue engineering.

Gene expression profiles of human chondrocytes during passaged monolayer cultivation.

Chondrocyte phenotype has been shown to dedifferentiate during passaged monolayer cultivation. Hence, we have investigated the expression profile of 27 chondrocyte-associated genes from both osteoarthritic cartilage tissue and healthy passaged human articular chondrocytes by quantitative real-time PCR. Our results indicate that the gene expression levels of matrix proteins and proteases in chondrocytes from monolayer culture decrease compared with those from cartilage tissue, while monolayer cultured chondrocytes from normal and osteoarthritic cartilage exhibit similar gene expression patterns. However, chondrocytic gene expression profiles were differentially altered at various stages of passage. The expression of the matrix proteins aggrecan, type II collagen, and fibromodulin inversely correlated with increasing passage number, while fibronectin and link protein exhibited a marked increase with passage. The expression of matrix proteinases MMP-3/9/13 and ADAMTS-4/5 decreased with passage, whereas proteinase inhibitors TIMP-2/3 were elevated. The cytokine IL-1 also showed increased expression with monolayer chondrocyte culture, while IGF-1 expression levels were diminished. No significant changes in TGF-beta, or the chondrogenic transcription factors Sox-9, c-fos, or c-jun were observed. Our data indicates that cultured chondrocytes undergo dedifferentiation during monolayer culture, although the gene expression level of transcription factors necessary for chondrogenesis remains unchanged. This data may prove important for the future development of more specific and efficacious cultivation techniques for human articular chondrocyte-based therapies.

Autologous tenocyte therapy using porcine-derived bioscaffolds for massive rotator cuff defect in rabbits.

Large and retracted rotator cuff tendon tears fail to repair or retear after surgical intervention. This study attempted to develop novel tissue-engineering approaches using tenocyte-seeded bioscaffolds for tendon reconstruction of massive rotator cuff tendon defect in rabbits. Porcine small intestine submucosa (Restore) and type I/III collagen bioscaffold (ACI-Maix) were chosen as bioscaffold carriers for autologous tenocytes. Biological characterization of autologous tenocytes was conducted before the implantation. The tenocyte-seeded bioscaffolds were implanted as interposition grafts to reconstruct massive rotator cuff tendon defects in rabbits. In situ reimplantation of the autologous rotator cuff tendon, excised during defect creation, served as a positive control. Histological outcomes were analyzed and semi-quantitatively graded at 4 and 8 weeks after surgery. At 4 weeks, both tenocyte-seeded bioscaffolds displayed inflammatory reaction similar to bioscaffold-only cuff reconstruction, and the histological grading were inferior to control repair. However, at 8 weeks, inflammatory reaction of both tenocyte-seeded bioscaffolds were dramatically less than with bioscaffold alone. In addition, bioscaffolds seeded with tenocytes generated a histological appearance similar to that of the positive control. The implantation of autologous tenocytes on collagen-based bioscaffolds results in better rotator cuff tendon healing and remodeling than with the implantation of bioscaffold alone.

Articular cartilage repair: procedures versus products.

This review discusses the current perspectives and practices regarding the treatment of articular cartilage injury. Specifically, the authors have delineated and examined articular cartilage repair techniques as either surgical procedures or manufactured products. Although both methodologies are used to treat articular cartilage injury, there are obvious advantages and disadvantages to the application of both, with the literature providing few recommendations on the most suitable regimen for the patient and surgeon. In recent times, cell-based tissue engineering products, predominantly autologous chondrocyte implantation, have been the subject of much research and have become clinically popular. Herein, we review the most used procedures and products in cartilage repair, compare and contrast their outcomes, and evaluate the issues that must be overcome in order to improve patient efficacy in the future.

Patellofemoral contact pressure following high tibial osteotomy: a cadaveric study.

Patella infera is a known complication of high tibial osteotomy (HTO) that can cause anterior knee pain due to excessive stresses associated with abnormal patellofemoral (PF) joint biomechanics. However, the translation of these abnormal biomechanics to native cartilage pressure has not been explored. The present study was designed to compare the PF contact pressures of three different HTOs in a human cadaveric model of valgus tibiofemoral correction. Nine fresh cadaveric knees underwent (1) medial opening wedge (OWHTO) with a proximal tuberosity osteotomy (PTO), (2) OWHTO with a distal tuberosity osteotomy (DTO), and (3) a lateral closing wedge (CWHTO). The specimens were mounted in a custom knee simulation rig, with muscle forces being simulated using a pulley system and weights. The PF contact pressure was recorded using an electronic pressure sensor at 15 degrees , 30 degrees , 60 degrees , 90 degrees , and 120 degrees of knee flexion, with results of the intact knees obtained as relative control. Compared to the intact knee, the DTO OWHTO and CWHTO did not significantly (P > 0.05) influence PF pressure at any flexion angle. On the other hand, PTO OWHTO lead to a significant elevation in PF cartilage pressure at 30 degrees (P < 0.05), 60 degrees (P < 0.005), and 90 degrees (P < 0.0005) knee flexion. We conclude from these results that DTO OWHTO maintains normal joint biomechanics and has no significant effect on PF cartilage pressure. In patients who complain of pre-existing anterior knee pain, DTO OWHTO or CWHTO should be considered.

Matrix-induced autologous chondrocyte implantation (MACI): biological and histological assessment.

Matrix-induced autologous chondrocyte implantation (MACI) has been a treatment of cartilage injury since 2000, but little is known of the histological paradigm of tissue regeneration after implantation. MACI is a stable cell-based delivery system that enables the regeneration of hyaline-like cartilage. From a cohort of 56 MACI patients, we examined the phenotype of chondrocytes seeded on type I/III collagen scaffold, and conducted progressive histologic assessment over a period of 6 months. Chondrocyte-seeded collagen scaffolds from patient implants were analyzed by electron microscopy, immunohistochemistry (type II collagen and S-100), and reverse transcription polymerase chain reaction (RT-PCR) (aggrecan and type II collagen). Coincidental cartilage biopsies were obtained at 48 hours, 21 days, 6 months, 8 months, 12 months, 18 months, and 24 months. Our data showed that chondrocytes on the collagen scaffold appeared spherical, well integrated into the matrix, and maintained the chondrocyte phenotype as evidenced by aggrecan, type II collagen, and S-100 expression. Progressive histologic evaluation of the biopsies showed the formation of cartilage-like tissue as early as 21 days, and 75% hyaline-like cartilage regeneration after 6 months. This preliminary study has suggested that MACI may offer an improved alternative to traditional treatments for cartilage injury by regenerating hyaline-like cartilage as early as 6 months after surgery.

The chondrocyte: biology and clinical application.

Chondrocyte is a unique cell type in articular cartilage tissue and is essential for cartilage formation and functionality. It arises from mesenchymal stem cells (MSCs) and is regulated by a series of cytokine and transcription factor interactions, including the transforming growth factor-beta super family, fibroblast growth factors, and insulin-like growth factor-1. To understand the biomechanisms of the chondrocyte differentiation process, various cellular model systems have been employed, such as primary chondrocyte culture, clonal normal cell lines (HCS-2/8, Ch-1, ATDC5, CFK-2, and RCJ3.1C5.18), and transformed clonal cell lines (T/C-28a2, T/C-28a4, C-28/I2, tsT/AC62, and HPV-16 E6/E7). Additionally, cell culture methods, including conventional monolayer culture, three-dimensional scaffold culture, bioreactor culture, pellet culture, and organ culture, have been established to create stable environments for the expansion, phenotypic maintenance, and subsequent biological study of chondrocytes for clinical application. Knowledge gained through these study systems has allowed for the use of chondrocytes in orthopedics for the treatment of cartilage injury and epiphyseal growth plate defects using tissue-engineering approaches. Furthermore, the potential of chondrocyte implantation for facial reconstruction, the treatment of long segmental tracheal defects, and urinary incontinence and vesicoureteral reflux are being investigated. This review summarizes the present study of chondrocyte biology and the potential uses of this cell in orthopedics and other disciplines.

Fibrin sealant promotes migration and proliferation of human articular chondrocytes: possible involvement of thrombin and protease-activated receptors.

Fibrin sealant (FS), a biological adhesive material, has been recently recommended as an adjunct in autologous chondrocyte implantation (ACI). While FS has been shown to possess osteoinductive potential, little is known about its effects on chondrogenic cells. In this study, we assessed the bioactivity of FS (Tisseel) on the migration and proliferation of human articular chondrocytes in vitro. Using a co-culture assay to mimic matrix-induced ACI (MACI), chondrocytes were found to migrate from collagen membranes towards FS within 12 h of culture, with significant migratory activity evident by 24 h. In addition, 5-bromo-2'-deoxyuridine (BrdU) incorporation experiments revealed that thrombin, the active component of the tissue glue, stimulated chondrocyte proliferation, with maximal efficacy observed at 48 h post-stimulation (1-10 U/ml). In an effort to elucidate the molecular mechanisms underlying these thrombin-induced effects, we examined the expression and activation of protease-activated receptors (PARs), established thrombin receptors. Using a combination of RT-PCR and immunohistochemistry, all four PARs were detected in human chondrocytes, with PAR-1 being the major isoform expressed. Moreover, thrombin and a PAR-1, but not other PAR-isotype-specific peptide agonists, were found to induce rapid intracellular Ca2+ responses in human chondrocytes in calcium mobilization assays. Together, these data demonstrate that FS supports both the migration and proliferation of human chondrocytes. We propose that these effects are mediated, at least in part, via thrombin-induced PAR-1 signalling in human chondrocytes.