Purpose We analyze the potential use of PRGF-Endoret® and human sera (HS) as a substitute of xenogeneic sera in corneal cell cultures. Methods Plasma rich in growth factors (PRGF) eye drop and HS was obtained at our local blood bank from human blood of healthy donors after informed consent. Briefly PRGF was purified using the Endoret kit in Ophthalmology. HS was obtained according to standardized protocols. Limbal stem cells were obtained from explants of 2-3mm in diameter of the limbal region of corneoscleral tissue and cultured in culture medium containing 10% PRGF-Endoret®, HS or fetal bovine serum (FBS) for 7 days. Afterwards, cells were fixed and growth area was quantified. Immunocytochemistry for p.63, ABCG-2 and cytokeratin was also performed in order to check their immunological markers. Results PRGF-Endoret®, HS and FBS, cell cultures showed a positive expression in p.63, ABCG-2 and cytokeratin. However some differences were found between cultures, limbal stem cell cultures supplemented with PRGF-Endoret® and HS showed a better growth area in contrast with the ones supplemented with FBS. Moreover, there was cellular growth in more of the explants cultured in PRGF-Endoret® and HS compared to FBS supplemented cultures. Conclusion PRGF-Endoret® and HS seems to improve the cellular growth and, while maintaining the p63 and ABCG-2 expression compared to FBS supplemented cultures. Therefore, PRGF-Endoret® or HS could be used as a substitute of xenogeneic sera, not only as an allogeneic product but also as an autologous product for the culture and expansion of corneal limbal stem cells.

Fetal bovine serum

10.1111/j.1755-3768.2014.s053.x

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Repair of Rat Mandibular Bone Defects by Alveolar Osteoblasts in a Novel Plasma-Derived Albumin Scaffold

Tissue Engineering Part A (2009)

Lorena Gallego Luís Junquera Eva Jover Verónica García María Álvarez‐Viejo

Repair of bone deficiencies in the craniofacial skeleton remains a difficult clinical problem. The aim of this study was to evaluate a novel albumin scaffold seeded with human alveolar osteoblasts and implanted into experimental mandibular defects. An experimental solid protein scaffold was prepared with human plasmatic albumin crossed with a glutaraldehyde-type agent. Microstructure of scaffold and mechanical properties were examined using scanning electron microscopy and a stress-controlled rheometer. Bilateral critical mandibular defects were created in eight immunodeficient rats. Defects of the right side of the mandibles received the cell-scaffold construct in all animals. All left mandibular defects were left untreated as blank controls. Sections of the defects were collected at 5, 8, and 11 weeks postsurgery and processed for histological and immunohistochemical observation, computed tomography examination, and computed tomography digital analysis. Histologically, bone formation was observed in both groups at 5 weeks postsurgery, and the engineered bone became more mature after 8 and 11 weeks, which was similar to normal bone. The origin of bone-forming cells within the defects and the localization of implanted human osteoblasts were confirmed by human vimentin expression. No bone formation could be observed at any control defect. Bone density after 8 weeks was significantly higher than that of the 5-week group (p = 0.02), and significant differences were also observed between 8 and 11 weeks (p < 0.01). The results indicate the clinical feasibility of albumin scaffold loaded with human alveolar cells and that it can be used as a good alternative for bone regeneration.

Bone remodeling

10.1089/ten.tea.2009.0517

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Multidisciplinary Treatment Outcome of Desmoid-Type Fibromatosis (Dtf). a Registry-Based Study from Spanish Group for Research on Sarcoma (Geis)

Annals of Oncology (2014)

Javier Martínez‐Trufero Rosa Álvarez Antonio López‐Pousa Isabel Sevilla P. Sancho Marquez

10.1093/annonc/mdu354.41

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Improved Tool for Predicting Skin Irritation on Reconstructed Human Epidermis Models Based on Electrochemical Impedance Spectroscopy

Biosensors (2023)

Manuel Chacón Natalia Vázquez Sergio Alonso‐Alonso Mairobi Persinal-Medina Sara Llames

The rabbit skin irritation test has been the standard for evaluating the irritation potential of chemicals; however, alternative methods that do not use animal testing are actively encouraged. Reconstructed human epidermis (RhE) models mimic the biochemical and physiological properties of the human epidermis and can be used as an alternative method. On RhE methods, the metabolic activity of RhE models is used to predict skin irritation, with a reduction in metabolic activity indicating a reduced number of viable cells and linking cell death to skin irritation processes. However, new challenges have emerged as the use of RhE models increases, including the need for non-invasive and marker-free methodologies to assess cellular states. Electrochemical impedance spectroscopy (EIS) is one such methodology that can meet these requirements. In this study, our results showed that EIS can differentiate between irritant and non-irritant chemicals, with a significant increase in the capacitance values observed in the irritant samples. A ROC curve analysis showed that the prediction method based on EIS met OECD TG 439 requirements at all time points and had 95% within-laboratory reproducibility. Comparison with the MTT viability assay showed that prediction using EIS achieved higher sensitivity, specificity, and accuracy. These results suggest that EIS could potentially replace animal testing in the evaluation of irritation potential and could be a valuable addition to in vitro testing strategies.

Skin irritation

Epidermis (zoology)

Viability assay

10.3390/bios13020162

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Low scale production of advanced therapy medicinal products for limbal stem cell deficiency

Álvaro Meana S Berisa Manuel Chacón Natalia Vázquez Mairobi Persinal Medina

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