title: Autologous chondrocytes as a novel source for neo-chondrogenesis in haemophiliacs.
abstract: Haemophilic arthropathy is the major cause of disability in patients with haemophilia and, despite prophylaxis with coagulation factor concentrates, some patients still develop articular complications. We evaluate the feasibility of a tissue engineering approach to improve current clinical strategies for cartilage regeneration in haemophiliacs by using autologous chondrocytes (haemophilic chondrocytes; HaeCs). Little is known about articular chondrocytes from haemophilic patients and no characterisation has as yet been performed. An investigation into whether blood exposure alters HaeCs should be interesting from the perspective of autologous implants. The typical morphology and expression of specific target genes and surface markers were therefore assessed by optical microscopy, reverse transcription plus the polymerase chain reaction (PCR), real-time PCR and flow-cytometry. We then considered chondrocyte behaviour on a bio-hybrid scaffold (based on polyvinyl alcohol/Wharton's jelly) as an in vitro model of articular cartilage prosthesis. Articular chondrocytes from non-haemophilic donors were used as controls. HaeC morphology and the resulting immunophenotype CD44(+)/CD49c(+)/CD49e(+)/CD151(+)/CD73(+)/CD49f(-)/CD26(-) resembled those of healthy donors. Moreover, HaeCs were active in the transcription of genes involved in the synthesis of the extracellular matrix proteins of the articular cartilage (ACAN, COL1A, COL2A, COL10A, COL9A, COMP, HAS1, SOX9), although the over-expression of COL1A1, COL10A1, COMP and HAS was observed. In parallel, the composite scaffold showed adequate mechanical and biological properties for cartilage tissue engineering, promoting chondrocyte proliferation. Our preliminary evidence contributes to the characterisation of HaeCs, highlighting the opportunity of using them for autologous cartilage implants in patients with haemophilia.
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Via Marzolo 5, 35131, Padua, Italy.|Foundation for Biology and Regenerative Medicine, Tissue Engineering and Signaling (TES) ONLUS, Padua, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Stocco
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Via Marzolo 5, 35131, Padua, Italy.|Foundation for Biology and Regenerative Medicine, Tissue Engineering and Signaling (TES) ONLUS, Padua, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Barbon
authors.affiliation: Haematology and Haemophilia Centre, Castelfranco Veneto, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Radossi
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Via Marzolo 5, 35131, Padua, Italy.
authors.forename: Senthilkumar
authors.initials: S
authors.lastname: Rajendran
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Via Marzolo 5, 35131, Padua, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Dalzoppo
authors.affiliation: Department of Surgery Oncology and Gastroenterology, University of Padua, Padua, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Bortolami
authors.affiliation: Department of Industrial Engineering, University of Padua, Padua, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bagno
authors.affiliation: Department of Women's and Children's Health, Pediatric Surgery, University of Padua, Padua, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Grandi
authors.affiliation: Department of Women's and Children's Health, Pediatric Surgery, University of Padua, Padua, Italy.
authors.forename: Pier Giorgio
authors.initials: PG
authors.lastname: Gamba
authors.affiliation: Foundation for Biology and Regenerative Medicine, Tissue Engineering and Signaling (TES) ONLUS, Padua, Italy.
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Parnigotto
authors.affiliation: Haematology and Haemophilia Centre, Castelfranco Veneto, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Tagariello
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Via Marzolo 5, 35131, Padua, Italy. claudio.grandi@unipd.it.|Foundation for Biology and Regenerative Medicine, Tissue Engineering and Signaling (TES) ONLUS, Padua, Italy. claudio.grandi@unipd.it.
authors.forename: Claudio
authors.initials: C
authors.lastname: Grandi
mesh_terms: D002358:Cartilage, Articular / Q000187:drug effects / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020219:Chondrogenesis / Q000187:drug effects; D055119:Elastic Modulus / Q000187:drug effects; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006467:Hemophilia A / Q000235:genetics / Q000473:pathology*; D006801:Humans; D016130:Immunophenotyping; D008297:Male; D008875:Middle Aged; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013314:Stress, Mechanical; D054457:Tissue Scaffolds; D014182:Transplantation, Autologous
chemical_list: D012333:RNA, Messenger; D011142:Polyvinyl Alcohol
keywords: Chondrocytes; Composite scaffold; Haemophilic arthropathy; Polyvinyl alcohol; Tissue engineering