Controlled expansion and differentiation of mesenchymal stem cells in a microcarrier based stirred bioreactor

Controlled expansion and differentiation of mesenchymal stem cells in a microcarrier based stirred bioreactor

The influence of HLA genotype on the development of metal hypersensitivity following joint replacement

\ua9 2022, The Author(s). Background: Over five million joint replacements are performed across the world each year. Cobalt chrome (CoCr) components are used in most of these procedures. Some patients develop delayed-type hypersensitivity (DTH) responses to CoCr implants, resulting in tissue damage and revision surgery. DTH is unpredictable and genetic links have yet to be definitively established. Methods: At a single site, we carried out an initial investigation to identify HLA alleles associated with development of DTH following metal-on-metal hip arthroplasty. We then recruited patients from other centres to train and validate an algorithm incorporating patient age, gender, HLA genotype, and blood metal concentrations to predict the development of DTH. Accuracy of the modelling was assessed using performance metrics including time-dependent receiver operator curves. Results: Using next-generation sequencing, here we determine the HLA genotypes of 606 patients. 176 of these patients had experienced failure of their prostheses; the remaining 430 remain asymptomatic at a mean follow up of twelve years. We demonstrate that the development of DTH is associated with patient age, gender, the magnitude of metal exposure, and the presence of certain HLA class II alleles. We show that the predictive algorithm developed from this investigation performs to an accuracy suitable for clinical use, with weighted mean survival probability errors of 1.8% and 3.1% for pre-operative and post-operative models respectively. Conclusions: The development of DTH following joint replacement appears to be determined by the interaction between implant wear and a patient’s genotype. The algorithm described in this paper may improve implant selection and help direct patient surveillance following surgery. Further consideration should be given towards understanding patient-specific responses to different biomaterials

Oxygen Tension Is a Determinant of the Matrix-Forming Phenotype of Cultured Human Meniscal Fibrochondrocytes

BACKGROUND: Meniscal cartilage displays a poor repair capacity, especially when injury is located in the avascular region of the tissue. Cell-based tissue engineering strategies to generate functional meniscus substitutes is a promising approach to treat meniscus injuries. Meniscus fibrochondrocytes (MFC) can be used in this approach. However, MFC are unable to retain their phenotype when expanded in culture. In this study, we explored the effect of oxygen tension on MFC expansion and on their matrix-forming phenotype. METHODOLOGY/PRINCIPAL FINDINGS: MFC were isolated from human menisci followed by basic fibroblast growth factor (FGF-2) mediated cell expansion in monolayer culture under normoxia (21%O(2)) or hypoxia (3%O(2)). Normoxia and hypoxia expanded MFC were seeded on to a collagen scaffold. The MFC seeded scaffolds (constructs) were cultured in a serum free chondrogenic medium for 3 weeks under normoxia and hypoxia. Constructs containing normoxia-expanded MFC were subsequently cultured under normoxia while those formed from hypoxia-expanded MFC were subsequently cultured under hypoxia. After 3 weeks of in vitro culture, the constructs were assessed biochemically, histologically and for gene expression via real-time reverse transcription-PCR assays. The results showed that constructs under normoxia produced a matrix with enhanced mRNA ratio (3.5-fold higher; p<0.001) of collagen type II to I. This was confirmed by enhanced deposition of collagen II using immuno-histochemistry. Furthermore, the constructs under hypoxia produced a matrix with higher mRNA ratio of aggrecan to versican (3.5-fold, p<0.05). However, both constructs had the same capacity to produce a glycosaminoglycan (GAG) -specific extracellular matrix. CONCLUSIONS: Our data provide evidence that oxygen tension is a key player in determining the matrix phenotype of cultured MFC. These findings suggest that the use of normal and low oxygen tension during MFC expansion and subsequent neo-tissue formation cultures may be important in engineering different regions of the meniscus

The effect of an autologous cellular gel-matrix integrated implant system on wound healing

<p>Abstract</p> <p>Background</p> <p>This manuscript reports the production and preclinical studies to examine the tolerance and efficacy of an autologous cellular gel-matrix integrated implant system (IIS) aimed to treat full-thickness skin lesions.</p> <p>Methods</p> <p>The best concentration of fibrinogen and thrombin was experimentally determined by employing 28 formula ratios of thrombin and fibrinogen and checking clot formation and apparent stability. IIS was formed by integrating skin cells by means of the <it>in situ </it>gelification of fibrin into a porous crosslinked scaffold composed of chitosan, gelatin and hyaluronic acid. The <it>in vitro </it>cell proliferation within the IIS was examined by the MTT assay and PCNA expression. An experimental rabbit model consisting of six circular lesions was utilized to test each of the components of the IIS. Then, the IIS was utilized in an animal model to cover a 35% body surface full thickness lesion.</p> <p>Results</p> <p>The preclinical assays in rabbits demonstrated that the IIS was well tolerated and also that IIS-treated rabbit with lesions of 35% of their body surface, exhibited a better survival rate (p = 0,06).</p> <p>Conclusion</p> <p>IIS should be further studied as a new wound dressing which shows promising properties, being the most remarkable its good biological tolerance and cell growth promotion properties.</p

Use of anodic oxidation to enhance the biocompatibility of tantalum implants

Tantalum (Ta) is increasingly being used in orthopaedic applications owing to its superior non-toxicity, fracture toughness, and corrosion resistance in comparison to titanium. The tantalum metal surface has an inherent oxide layer similar to what is observed in titanium and its alloys. However, the surface topology and chemistry of this layer are not conducive to integration to the bone; thus alteration is required to prevent joint failure. The topology of the Ta implant surface can be modified by anodisation in an acid electrolyte to produce a porous and rough coating. Furthermore, the surface chemistry and resultant biologicalresponse of these coatings can be modified by incorporating specific ions on the surface. This can be either done through the selection of the appropriate electrolyte or by ion implantation on the coating surface. Moreover, the oxide layer formed (i.e., Ta2O5) is expected to have photocatalytic properties and it is possible to activate them to produce ananti-microbial effect using X-rays, as has been observed forTiO2 coatings on Ti6Al4V implants. The present work investigates the effects of the anodization parameters and postanodisationtreatment on the characteristics of the coatings formed by anodization, and their impact on osteoblast growth

Anti-Microbial Bioceramic Coatings on Ti6Al4V Surfaces by Anodisation

Titanium and its alloys are a popular choice for biomedical implants, particularly knee and hip replacements, owing to their high biocompatibility and superior mechanical properties. The insertion of these implants is done through surgery and the resulting incisions provide an entry point for microbes, thereby increasing the risk of infections andrejection responses. These infections are difficult to treat and impact severely on patient health and implant lifetime. A potential solution is the fabrication of anti-microbial coatings of TiO2 on the implant surface which are photocatalytic and can be activated by X-rays to provide an anti-microbial effect