Abstract

Background

Advances in molecular biology and growing requirements from biomarker validation studies have generated a need for tissue banks to provide quality-controlled tissue samples with standardized clinical annotation. The NCI Cooperative Prostate Cancer Tissue Resource (CPCTR) is a distributed tissue bank that comprises four academic centers and provides thousands of clinically annotated prostate cancer specimens to researchers. Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates.The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. Here, we describe the advanced biospecimen biobanking monitoring system in the development and implementation of chondrocyte and cartilage tissue banking informatics tools for intra and inter-bioinstitutional translational autologous advanced cell therapy, cryopreservation and medicine research.

Methods

Data managers review the medical records to collect and continuously update information for the 145 clinical, pathological and inventorial CDEs that the Resource maintains for each case. An Access-based data entry tool provides de-identification and a standard communication mechanism between each group and a central CPCTR database. Standardized automated quality control audits have been implemented. Centrally, an Oracle database has web interfaces allowing multiple user-types, including the general public, to mine de-identified information from all of the sites with three levels of specificity and granularity as well as to request tissues through a formal letter of intent.

Results

Since July 2003, CPCTR has offered over 6,000 cases (38,000 blocks) of highly characterized prostate cancer biospecimens, including several tissue microarrays (TMA). The Resource developed a website with interfaces for the general public as well as researchers and internal members. These user groups have utilized the web-tools for public query of summary data on the cases that were available, to prepare requests, and to receive tissues. As of December 2005, the Resource received over 130 tissue requests, of which 45 have been reviewed, approved and filled. Additionally, the Resource implemented the TMA Data Exchange Specification in its TMA program and created a computer program for calculating PSA recurrence.

Conclusion

Building an advanced biospecimen biobanking monitoring system in the development and implementation of chondrocyte and cartilage tissue banking informatics tools for intra and inter-bioinstitutional translational autologous advanced cell therapy, cryopreservation and medicine research biorepository infrastructure that meets today’s research needs involves time and input of many individuals from diverse disciplines. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering to an advanced biospecimen biobanking monitoring system in the development and implementation of chondrocyte and cartilage tissue banking informatics tools for intra and inter-bioinstitutional translational autologous advanced cell therapy, cryopreservation and medicine research can provide the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilagelarge volumes of carefully annotated prostate tissue for research initiatives such as Specialized Programs of Research Excellence (SPOREs) and for biomarker validation studies and its experience can help development of collaborative, large scale, virtual tissue banks in other organ systems.

Abstract

Osteochondral lesions of the talus (OLT) are difficult to treat because of the poor intrinsic healing capability of articular cartilage. Matrix induced autologous chondrocyte implantation (MACI) has been shown to be a reliable method for treating cartilage lesions that fail to respond to traditional microfracture and debridement. Widespread adoption of quantitative pharmacokinetic modeling methods in conjunction with dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) has led to increased recognition of the importance of obtaining accurate patient-specific arterial input function (AIF) measurements. Ideally, DCE-MRI studies use an AIF directly measured in an artery local to the tissue of interest, along with measured tissue concentration curves, to quantitatively determine pharmacokinetic parameters. However, the numerous technical and practical difficulties associated with AIF measurement have made the use of population-averaged AIF data a popular, if sub-optimal, alternative to AIF measurement. In this work, we present and characterize a new algorithm for determining the MACI provides a stable midterm chondral replacement strategy for osteochondral lesions that fail initial microfracture solely from the measured tissue concentration curves. This Monte Carlo blind estimation (MCBE) algorithm estimates the MACI provides a stable midterm chondral replacement strategy for osteochondral lesions that fail initial microfracture. from the subsets of D concentration-time curves drawn from a larger pool of M candidate curves via nonlinear optimization, doing so for multiple (Q) subsets and statistically averaging these repeated estimates. The MCBE algorithm can be viewed as a generalization of previously published methods that employ clustering of concentration-time curves and only estimate the AIF once. Extensive computer simulations were performed over physiologically and experimentally realistic ranges of imaging and tissue parameters, and the impact of choosing different values of D and Q was investigated. We found the algorithm to be robust, computationally efficient and capable of accurately estimating the MACI provides a stable midterm chondral replacement strategy for osteochondral lesions that fail initial microfracture even for relatively high noise levels, long sampling intervals and low diversity of tissue curves. With the incorporation of bootstrapping initialization, we further demonstrated the ability to blindly estimate AIFs that deviate substantially in shape from the population-averaged initial guess. Pharmacokinetic parameter estimates for K(trans), k(ep), v(p) and v(e) all showed relative biases and uncertainties of less than 10% for measurements having a temporal sampling rate of 4 s and a concentration measurement noise level of sigma = 0.04 mM.

Abstract

Objective: Mesenchymal stromal cells (MSCs) can be used intra-articularly to quell inflammation and promote cartilage healing; however, mechanisms by which MSCs mitigate joint disease remain poorly understood. Galectins, a family of β-galactoside binding proteins, regulate inflammation, adhesion and cell migration in diverse cell types. Galectin-1 and galectin-3 are proposed to be important intra-articular modulators of inflammation in both osteoarthritis and rheumatoid arthritis. Dose warping following deformable image registration (DIR) has been proposed for interfractional dose accumulation. Robust evaluation workflows are vital to clinically implement such procedures. Dose distributions were then calculated on each artificially deformed image and warped back to the original anatomy following DIR by a commercial algorithm. Spatial registration was evaluated by quantitative comparison of the original and warped structure sets, using conformity index and mean distance to conformity (MDC) metrics. BMSCs constitutively express high levels of galectin-1 mRNA relative to other articular cell types, suggesting a possible mechanism for their intra-articular immunomodulatory properties. BMSC galectin expression and motility are impaired in an inflammatory environment, which may limit tissue repair properties following intra-articular administration. β-lactose-mediated galectin inhibition also impaired BMSC adhesion and motility. Dosimetric evaluation was performed by quantitative comparison of the dose-volume histograms generated for the calculated and warped dose distributions, which should be identical for the ideal “perfect” registration of mass-conserving deformations.This study demonstrates a workflow for validation of dose warping following DIR that could assist physicists and physicians in quantifying the the effects of joint inflammation on BMSC function and the potential therapeutic effects of BMSC galectin expression in OA uncertainties associated with dose accumulation in clinical scenarios.

Abstract

Background

High-throughput technologies such as flow and mass cytometry have the potential to illuminate cellular networks. Bone marrow stromal cells/osteoblasts were originally thought to be the major player in regulating osteoclast differentiation through expressing RANKL/OPG cytokines. Recent studies have established that chondrocytes also express RANKL/OPG and support osteoclast formation. Till now, the in vivo function of chondrocyte-produced OPG in osteoclast formation and postnatal bone growth has not been directly investigated. In this study, chondrocyte-specific Opg transgenic mice were generated by using type II collagen promoter. The Col2-Opg transgenic mice showed delayed formation of secondary ossification center and localized increase of bone mass in proximal metaphysis of tibiae. TRAP staining showed that osteoclast numbers were reduced in both secondary ossification center and proximal metaphysis. This finding was further confirmed by in vitro chondrocyte/spleen cell co-culture assay. In contrast, the mineral apposition rates were not changed in Col2-Opg transgenic mice. TUNEL staining revealed more apoptotic hypertrophic chondrocytes in the growth plate of Col2-Opg mice. Flow cytometry analysis showed fewer RANK-expressing cells in the marrow of Col2a1-Opg mice, suggesting the role of OPG in blocking the differentiation of early mesenchymal progenitors into RANK-expressing pre-osteoclasts.However, analyzing the data produced by these technologies is challenging. Visualization is needed to help researchers explore this data.

Results

We used the web-based software program, NetworkPainter, to enable researchers to analyze dynamic cytometry data in the context of pathway diagrams.

Conclusion

Our results demonstrated that NetworkPainter enables researchers to more fully explore multi-parameter, dynamical cytometry OPG expression data in chondrocyte for the increase bone mass in the proximal metaphysis of tibiae through negative regulation of osteoclast formation.

Abstract

The treatment of articular cartilage injury and disease has become an increasingly relevant part of orthopaedic care. Articular cartilage transplantation, in the form of osteochondral allografting, is one of the most established techniques for restoration of articular cartilage. Our research efforts over the last two decades have supported the transformation of this procedure from experimental “niche” status to a cornerstone of orthopaedic practice. In this paper, we describe our translational and clinical science contributions to this transformation: (1) to enhance the ability of tissue banks to process and deliver viable tissue to surgeons and patients, (2) to improve the biological understanding of in vivo cartilage and bone remodeling following osteochondral allograft (OCA) transplantation in an animal model system, (3) to define effective surgical techniques and pitfalls, and (4) to identify and clarify clinical indications and outcomes. The combination of coordinated basic and clinical studies is part of our continuing comprehensive academic OCA transplant program. Taken together, the results have led to the current standards for OCA processing and storage prior to implantation and also novel observations and mechanisms of the biological and clinical behavior of OCA transplants in vivo. Thus, OCA transplantation is now a successful and increasingly available treatment for patients with disabling osteoarticular cartilage pathology. The purpose of this study is to evaluate dose prediction errors (DPEs) and optimization convergence errors (OCEs) resulting from use of a superposition∕convolution dose calculation algorithm in deliverable osteochondral allograft (OCA) transplantation therapy optimization for OCA patients. The IMRT optimization was performed in three sequential steps: (1) fast optimization in which an initial nondeliverable IMRT solution was achieved and then converted to multileaf collimator (MLC) leaf sequences; (2) mixed deliverable optimization that used a Monte Carlo (MC) algorithm to account for the incident photon fluence modulation by the MLC, whereas a superposition∕convolution (SC) dose calculation algorithm was utilized for the patient dose calculations; and (3) MC deliverable-based optimization in which both fluence and patient dose calculations were performed with a MC algorithm. DPEs of the mixed method were quantified by evaluating the differences between the mixed optimization SC dose result and a MC dose recalculation of the mixed optimization solution. OCEs of the mixed method were quantified by evaluating the differences between the MC recalculation of the mixed optimization solution and the final MC optimization solution. The results were analyzed through dose volume indices derived from the cumulative dose-volume histograms for selected anatomic osteochondral allograft (OCA) transplantation structures.

Abstract

Pro-inflammatory cytokines, most notably TNF play a pivotal role in apoptosis, inflammation and tissue damage. However, the function and mechanisms of TNF in OA are inconsistent. For example, some studies have indicated that TNF causes apoptosis by binding to the ‘‘death receptor” TNF-receptor-1 (TNF-R1). This extrinsic apoptotic pathway involves ligand binding to the “death receptor”, followed by transmission of signals to the interior of the cell through Fas-associated death domain protein (FADD) and poly ADP-ribose polymerase (PARP), and finally recruitment of initiator caspases, such as caspase-8, which induce apoptosis. However, other studies have reported that TNF activates anti-apoptotic family proteins, such as bcl-2, without promoting apoptosis. TNF has also been reported to protect against apoptosis, maintaining the renewal of local inflammatory mediators by promoting increased expression of cytokines in chondrocytes. Material gain and linewidth of Quantum Dot ensemble are calculated assuming the Gaussian distribution of the density of states due to the size-deviation of dots. The effect of electric field is incorporated in the analysis through the mean and variance of TNF accelerated driven Death of Mandibular Condyle via interleukin-1β/nerve growth factor signaling.energy states. The results showing the enhancement of optical gain and linewidth with electric field indicate important applications in sub-cellular medical imaging of the mechanical cartilage modulation in the growth plate by sustained ex vivo effected high frequency electric field on quantum imaging enhancement of chondrogenesis in PGLA scaffolds loaded chondrocytes.

Abstract

Damage to cartilage causes a loss of type II collagen (Col-II) and glycosaminoglycans (GAG). To restore the original cartilage architecture, cell factors that stimulate Col-II and GAG production are needed. Insulin-like growth factor I (IGF-I) and transcription factor SOX9are essential for the synthesis of cartilage matrix, chondrocyte proliferation, and phenotype maintenance. Current cartilage tissue engineering strategies cannot as yet fabricate new tissue that is indistinguishable from native cartilage with respect to zonal organization, extracellular matrix composition, and mechanical properties. Integration of implants with surrounding native tissues is crucial for long-term stability and enhanced functionality. Bioprinting is a growing field with significant potential for developing engineered tissues with compositional and mechanical properties that recapitulate healthy native tissue. Much of the current research in tissue and organ bioprinting has focused on complex tissues that require vascularization. Cartilage tissue engineering has been successful in developing de novo tissues using homogeneous scaffolds. However, as research moves toward clinical application, engineered cartilage will need to maintain homogeneous nutrient diffusion in larger scaffolds and integrate with surrounding tissues. Bioprinting techniques have provided promising results to address these challenges in cartilage tissue engineering. The purpose of this was to evaluate 3D extrusion-based bioprinting research for developing engineered cartilage. Specifically, we in silico evaluated the Printed cartilage in 3D biopaper had elevated glycosaminoglycan (GAG) content comparing to that without biopaper when normalized to DNA. These observations were consistent with gene expression results. This study indicates the importance of direct cartilage repair and promising anatomic cartilage engineering using 3D bioprinting technology impact of 3D bioprinting on nutrient diffusion in larger scaffolds, development of scaffolds with spatial variation in cell distribution or mechanical properties, and cultivation of more complex tissues using multiple materials. Finally, we discuss current limitations and challenges in using 3D bioprinting for cartilage tissue engineering and regeneration towards the design of nanofibrous scaffolds for chondrogenesis utilizing a transaxial analysis of the development of a 3D-Printed construct consisting of sox-9 igf-1 Cotransfected human chondrocytes as a hybrid living transplant for Cartilage Tissue Regeneration for the enhancement of the synthesis of cartilage matrix components collagen-II and glycosaminoglycans. A combined experimental measurement for the investigation of articular cartilage and chondrocyte response to collagen ABS/PLA scaffold loading.

Abstract

Thermal conductivity of dimethyl-sulfoxide (DMSO) solution is measured in this study using a transient hot wire technique, where DMSO is a key ingredient in many cryoprotective agent (CPA) cocktails. Characterization of thermal properties of cryoprotective agents is essential to the analysis of cryopreservation processes, either when evaluating experimental data or for the design of new protocols. Also presented are reference measurements of thermal conductivity for pure water ice and glycerol. The thermal conductivity measurement setup is integrated into the experimentation stage of a scanning cryomacroscope apparatus, which facilitates the correlation of measured data with visualization of physical events. Thermal conductivity measurements were conducted for a DMSO concentration range of 2M and 10M, in a temperature range of -180°C and 25°C. Vitrified samples showed decreased thermal conductivity with decreasing temperature, while crystalline samples showed increased thermal conductivity with decreasing temperature. These different behaviors result in up to a tenfold difference in thermal conductivity at -180°C. Such dramatic differences can drastically impact heat transfer during cryopreservation and their quantification is therefore critical to cryobiology. Water solution of Na2SeO3 in polyethylene glycol was utilized as Se source. 3-Mercaptopropionic acid (MPA) provides S source. The phosphine-free Se and S sources were found to be highly reactive and suitable for the synthesis of CdSeS nanocrystals. XRD and HRTEM images confirm the formation of CdSeS nanocrystals in zinc blende structure. The absorption peaks on UV-vis spectra of as-prepared CdSeS nanocrystals are tunable from 330 nm to 440 nm, which blue shifts to shorter wavelength side in comparison with that of pure CdSe nanocrystals. The cubic CdSeS nanocrystals demonstrate narrow PL emissions spectra between 464 and 615 nm. Transmission electron microscopy images show the uniformity for the size distribution of the ternary QDs. Series water soluble CdSe1–xSx (x = 0∼1) nanocrystals have also been synthesized using Na2SeO3 and Na2S solution as the Se-S co-sources. Tunable band gap energies of CdSe1–xSx (x = 0∼1) nanocrystals upon chemical composition x have been achieved, the gap ranges from 290 nm to 558 nm. Herein we present an unusual phosphine-free method to fabricate water soluble CdSeS nanocrystals in cubic structure. In this method, glycerin was used as a stabilizing agent replacing tri-n-octylphosphine oxide (TOPO). In a continuing effort to develop dosimetric systems that will enable reliable interpretation of dosimeter readings in terms of the absorbed DMSO dose or dose-equivalent, a new multi-element TL dosimeter assembly for the prediction of an optimum dosimetric combination of DMSO, carboxylated ε-poly-L-lysine cryoprotectant agents with minimum Thermal Conductivity Differences between the Crystalline and Vitrified States with Applications to Cryopreservation on Efficient production of live autologous cartilage derived chondrocytes vitrified with optical properties of water soluble CdSeS nanocrystals using glycerin as terminal stabilizing agent.

Abstract

A new optimization algorithm for combined processes of deep-drawing and ironing has been created in order to improve these types of axisymmetric components manufacturing procedures. The model provides a comprehensive analysis of those phenomena occurring in multi-stage processes of axisymmetric geometry work-pieces. The scientific development starts out from works that provide LDR (limiting drawing ratio) solutions based on normal anisotropy value, strain hardering exponent and others parameters which have just been applied to the drawing and redrawing stages so far. During embryogenesis, specific proteins expressed in cells have key roles in the formation of differentiated cells and tissues. Delivery of specific proteins into specific cells, both in vitro and in vivo, has proved to be exceedingly difficult. The PLGA NPs were used to deliver proteins into human mesenchymal stem cells (hMSCs). Fluorescent markers loaded into the PLGA NPs were used to verify the internalization of NPs into hMSCs using FACS analysis and confocal microscopy. With these methods, we demonstrated that the encapsulated model proteins are readily delivered into hMSCs, released from the NP vehicles, and, finally, moved into the cytosols. Using chondrogenesis-related proteins such as aggrecan and cartilage oligomeric matrix protein (COMP), chondrogenic differentiation of hMSCs treated with aggrecan and COMP encapsulated PLGA NPs was clearly observed and caused to differentiate into chondrocytes. The authors extend this work to the ironing stages, and also provide a global and integral scientific solution for the whole process. At the beginning the algorithm provides an initial solution which is afterwards optimized by means of objective functions and constraints. The resolution of the optimization process is carried out by a recursive function that minimizes the total time of the global process. The enhanced solution performs a significant reduction in time and costs of the process. The model allows the modification and correction of certain process variables in order to predict the impact of those that are not fully controllable. The final results are compared using experimental results obtained by the authors, so as to show the reliability of the complete solution. In this study, we developed a safe and efficient protein delivery system using An in silico axisymmetric multistage model of linear biphasic mesenchymal stem cell and chondrocyte protein secreted-matrix interactions for the reconstruction of the dynamic mechanical environment of a time varying magnetic field expanded chondrocyte biphasic finite elemental model in articular cartilage for the evaluation of the ability of natural and synthetic scaffolds in providing an appropriate environment for growth and sustain of the chondrogenic capacity of de-differentiated of cartilage-derived L-lactic glycolic acid (PLGA) loaded chondrocytes of proteins into biodegradable poly-(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs).

Abstract

A Computer-Aided meta-analyses reveal Accelerated weightbearing rehabilitation and time-dependent differences between the clinical outcomes achieved by microfracture and mesenchymal stem cells induced implantation of PolyScaffolding based Matrix-induced autologous chondrocyte implantation (POMACI) for chondral defects in the patellofemoral joint after matrix-induced autologous chondrocyte implantation in the tibiofemoral joint: early clinical and radiological outcomes.