Characterization of the molecular composition and in vitro regenerative capacity of platelet-based bioproducts and related subfractions.

The use and demand of platelet-based bioproducts in regenerative medicine is steadily increasing. However, it is very difficult to establish the real clinical benefits of these therapies, as the lack of characterization and detailed production methods of platelet-based bioproducts persists in the literature and precludes cross-study comparisons. We characterized the molecular composition and in vitro regenerative capacity of platelet-rich plasma (PRP) produced in a closed-system. Furthermore, we performed a parallel characterization on different PRP subfractions (plasma and plasma-free platelet lysate), identifying that the fractions containing platelet-derived cargo exert the most potent regenerative capacity. This observation led us to develop a method to obtain a platelet secretome highly enriched in growth factors, free of plasma and cellular components (PCT/IB2022/057936), with the aim of establishing a superior bioproduct. The molecular characterization of secretomes revealed agonist-dependent differences, which correlates with beneficial grades of regenerative capacity. Importantly, secretomes showed general superiority to PRP in vitro. We discuss the variables influencing the bioproduct quality (inter-donor variation, platelet source and processing methods). Finally, we propose that the characteristics of secretomes circumvents certain limitations of PRP (autologous vs allogeneic), and envision that optimizing post-processing protocols (nanoencapsulation, lyophilization), would allow their clinical application even beyond regenerative medicine. STATEMENT OF SIGNIFICANCE: The use and demand of platelet-based bioproducts in regenerative medicine is steadily increasing. However, it is very difficult to establish the real clinical benefits of these therapies, or to improve/personalize them, as the lack of characterization of the bioproducts and their production methods is a constant in the literature, reason that precludes cross-study comparisons. In the present manuscript, we provide a comprehensive molecular and functional characterization of platelet-based bioproducts and subfractions, including platelet rich plasma, plasma fractions and platelet secretomes produced with a methodology developed by our group. Our results show that the molecular composition of each fraction correlates with its regenerative capacity in vitro. Thus, a rigorous characterization of platelet-derived bioproducts will potentially allow universal use, customizing and new applications.

Clinical Response After Treatment of Knee Osteoarthritis With a Standardized, Closed-System, Low-Cost Platelet-Rich Plasma Product: 1-Year Outcomes.

Background: Intra-articular infiltration of platelet-rich plasma (PRP) is an alternative therapeutic option to classic hyaluronic acid for the treatment of symptomatic knee osteoarthritis (KOA). However, variation in preparation methods and quality assessment of PRP makes the study of its real clinical efficacy difficult. Purpose: To (1) evaluate the clinical efficacy of a characterized PRP product prepared in a standardized manner and in a closed-system for the treatment of KOA and to (2) evaluate the association of the clinical response to PRP-related variables. Study Design: Case series; Level of evidence, 4. Methods: We recruited 130 patients with nonoperative KOA and evaluated them for 1 year. PRP was prepared from a donation of autologous blood, obtaining 3 aliquots of approximately 10mL of product, which were frozen, allowing platelet disruption, platelet factor release, and long-term storage, until administration. Patients were treated 3 consecutive times every 4 weeks with an intra-articular PRP knee injection under sterile conditions. Complete blood count was performed on the whole-blood sample and the processed PRP before freezing it, for product quality assessment. Patients were assessed using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and basic satisfaction scale at 3 months, 6 months, and 1 year after intervention. Results: Quality assessment confirmed a leukocyte-poor PRP product (white blood cell count, 0.09 ± 0.09 × 109/L) with a high platelet purity (platelet count, 630.86 ± 191.75 × 109/L). WOMAC scores improved, and basic satisfaction was achieved in 70% of patients. No adverse events were reported. No correlations were observed between PRP quality parameters and clinical results. PRP complete treatment production costs were €108/US$125 (€36/US$41.6 per injection). Conclusion: This standardized PRP production method resulted in improved WOMAC scores at 1 year postoperatively in 70% of patients with KOA. This technique was safe and affordable and ensured consecutive infiltrations with the same product to each patient.

Platelet-derived bio-products: Classification update, applications, concerns and new perspectives.

Platelet derived bio-products in the form of platelet rich plasma, plasma rich in growth factors, or plasma-free platelet releasates, are being studied worldwide with the aim of proving their efficacy in tissue regeneration within many different clinical areas, such as traumatology, maxillofacial surgery, ophthalmology, dermatology and otorhinolaryngology, amongst others. The current lack of consensus in the preparation method and application form, or in the quality assessment of each bio-product, precludes adequate interpretation of the relevance of reported clinical outcomes, and, while many in clinicians are very positive about them, many are sceptic. Relevant aspects of these products are considered to propose a classification nomenclature which would aid a comprehensive comparison of clinical outcomes of bio-products of the same characteristics. Finally, the uses of platelet-derived bio-products in in vitro culture (for cell therapy purposes) as a substitute of animal-origin sera, and other future perspectives of applications of platelet-derived bio-products are discussed.

Diagnóstico precoz de amiloidosis sistémica mediante biopsia de ligamento transverso del carpo durante la cirugía del síndrome del túnel carpiano / Early diagnosis of systemic amyloidosis by means of a transverse carpal ligament biopsy carried out during carpal tunnel syndrome surgery

Introducción y objetivo. Planteamos la posibilidad de que el análisis sistemático de una muestra de ligamento anular anterior del carpo (LAAC) obtenida durante la cirugía rutinaria de síndrome del túnel carpiano (STC) pueda constituir un método de diagnóstico precoz para la amiloidosis sistémica. Material y métodos. Estudio prospectivo en el que se recogieron las muestras consecutivas de LAAC de 147 pacientes intervenidos por STC en el Hospital Universitario de León entre abril de 2006 y mayo de 2007. En aquellos en los que se observó depósito de amiloide en la muestra de LAAC, se completó el estudio con la realización de punción-aspiración con aguja fina (PAAF) de grasa abdominal subcutánea, utilizando en ambos casos la tinción de Rojo Congo. Los casos positivos fueron derivados a los servicios de Medicina Interna y/o Hematología, observando su evolución durante 8 años. Resultados. Se observó depósito de amiloide en LAAC en 29 pacientes (19,7%), y pudo realizarse PAAF de grasa abdominal en 19 de ellos (65,5%), resultando positiva en 11 (57,9%); de ellos, 4 pacientes (3% del total) desarrollaron en los 3 años posteriores episodios atribuibles a la amiloidosis. Conclusiones. La biopsia rutinaria de LAAC durante la cirugía de STC podría adelantar el diagnóstico de la amiloidosis sistémica (AU)

Introduction and objective. The systematic analysis of a carpal transverse ligament (CTL) sample obtained during routine carpal tunnel syndrome (CTS) surgery may constitute a method of early diagnosis for systemic amyloidosis. Material and methods. Prospective study carried out on 147 consecutive CTL samples collected from patients intervened for CTS at the University Hospital of León from April 2006 to May 2007. In those cases in which amyloid deposition was observed in the CTL sample, the study was completed with a fine needle aspiration biopsy (FNAB) of the subcutaneous fascia, using the Red Congo stain in both cases. Positive cases were referred to the Internal Medicine and/or Hematology departments, and their evolution was monitored for up to 8 years. Results. CTL amyloid deposition was observed in 29 patients (19.7%), with a FNAB only being performed in 19 of them (65.5%). The test was positive in 11 cases (57.9%), and 4 patients in this subgroup (3% of the total) developed events attributable to amyloidosis over the following 3 years. Conclusions. A CTL routine biopsy carried out during CTS surgery may anticipate the systemic amyloidosis diagnosis (AU)

Early diagnosis of systemic amyloidosis by means of a transverse carpal ligament biopsy carried out during carpal tunnel syndrome surgery. / Diagnóstico precoz de amiloidosis sistémica mediante biopsia de ligamento transverso del carpo durante la cirugía del síndrome del túnel carpiano.

INTRODUCTION AND OBJECTIVE: The systematic analysis of a carpal transverse ligament (CTL) sample obtained during routine carpal tunnel syndrome (CTS) surgery may constitute a method of early diagnosis for systemic amyloidosis. MATERIAL AND METHODS: Prospective study carried out on 147 consecutive CTL samples collected from patients intervened for CTS at the University Hospital of León from April 2006 to May 2007. In those cases in which amyloid deposition was observed in the CTL sample, the study was completed with a fine needle aspiration biopsy (FNAB) of the subcutaneous fascia, using the Red Congo stain in both cases. Positive cases were referred to the Internal Medicine and/or Hematology departments, and their evolution was monitored for up to 8 years. RESULTS: CTL amyloid deposition was observed in 29 patients (19.7%), with a FNAB only being performed in 19 of them (65.5%). The test was positive in 11 cases (57.9%), and 4 patients in this subgroup (3% of the total) developed events attributable to amyloidosis over the following 3 years. CONCLUSIONS: A CTL routine biopsy carried out during CTS surgery may anticipate the systemic amyloidosis diagnosis.