PHB/CHIT Scaffold as a Promising Biopolymer in the Treatment of Osteochondral Defects—An Experimental Animal Study

Biopolymer composites allow the creation of an optimal environment for the regeneration of chondral and osteochondral defects of articular cartilage, where natural regeneration potential is limited. In this experimental study, we used the sheep animal model for the creation of knee cartilage defects. In the medial part of the trochlea and on the medial condyle of the femur, we created artificial defects (6 × 3 mm2) with microfractures. In four experimental sheep, both defects were subsequently filled with the porous acellular polyhydroxybutyrate/chitosan (PHB/CHIT)-based implant. Two sheep had untreated defects. We evaluated the quality of the newly formed tissue in the femoral trochlea defect site using imaging (X-ray, Computer Tomography (CT), Magnetic Resonance Imaging (MRI)), macroscopic, and histological methods. Macroscopically, the surface of the treated regenerate corresponded to the niveau of the surrounding cartilage. X-ray examination 6 months after the implantation confirmed the restoration of the contour in the subchondral calcified layer and the advanced rate of bone tissue integration. The CT scan revealed a low regenerative potential in the bone zone of the defect compared to the cartilage zone. The percentage change in cartilage density at the defect site was not significantly different to the reference area (0.06–6.4%). MRI examination revealed that the healing osteochondral defect was comparable to the intact cartilage signal on the surface of the defect. Hyaline-like cartilage was observed in most of the treated animals, except for one, where the defect was repaired with fibrocartilage. Thus, the acellular, chitosan-based biomaterial is a promising biopolymer composite for the treatment of chondral and osteochondral defects of traumatic character. It has potential for further clinical testing in the orthopedic field, primarily with the combination of supporting factors.

[1]  M. Pušić,et al.  Characterization of Chitosan-Based Scaffolds Seeded with Sheep Nasal Chondrocytes for Cartilage Tissue Engineering , 2021, Annals of Biomedical Engineering.

[2]  R. Burgkart,et al.  Thickness of the Stifle Joint Articular Cartilage in Different Large Animal Models of Cartilage Repair and Regeneration , 2020, Cartilage.

[3]  Gelmires de Araújo Neves,et al.  A Review on Chitosan’s Uses as Biomaterial: Tissue Engineering, Drug Delivery Systems and Cancer Treatment , 2020, Materials.

[4]  Chris Peach,et al.  Biological perspectives and current biofabrication strategies in osteochondral tissue engineering , 2020, Biomanufacturing Reviews.

[5]  S. Sim,et al.  Guided bone marrow stimulation for articular cartilage repair through a freeze-dried chitosan microparticle approach , 2020 .

[6]  Md. Shahruzzaman,et al.  Chitosan based bioactive materials in tissue engineering applications-A review , 2020, Bioactive materials.

[7]  Ž. Knez,et al.  Poly(3-hydroxybutyrate): Promising biomaterial for bone tissue engineering , 2019, Acta pharmaceutica.

[8]  M. Alini,et al.  Articular fibrocartilage - Why does hyaline cartilage fail to repair? , 2019, Advanced drug delivery reviews.

[9]  R. Búreš,et al.  Polyhydroxybutyrate/Chitosan 3D Scaffolds Promote In Vitro and In Vivo Chondrogenesis , 2019, Applied Biochemistry and Biotechnology.

[10]  M. Marcacci,et al.  A Composite Chitosan-Reinforced Scaffold Fails to Provide Osteochondral Regeneration , 2019, International journal of molecular sciences.

[11]  L. Alexopoulos,et al.  An ex vivo tissue model of cartilage degradation suggests that cartilage state can be determined from secreted key protein patterns , 2019, bioRxiv.

[12]  Ľ. Medvecký,et al.  The Serum Protein Profile and Acute Phase Proteins in the Postoperative Period in Sheep after Induced Articular Cartilage Defect , 2019, Materials.

[13]  C. Scirè,et al.  Osteoarthritis and its management - Epidemiology, nutritional aspects and environmental factors. , 2018, Autoimmunity reviews.

[14]  F. Guilak,et al.  Osteoarthritis as a disease of the cartilage pericellular matrix. , 2018, Matrix biology : journal of the International Society for Matrix Biology.

[15]  D. Lieberman,et al.  Modern-day environmental factors in the pathogenesis of osteoarthritis , 2018, Nature Reviews Rheumatology.

[16]  Fubo Chen,et al.  Animal Models Used for Testing Hydrogels in Cartilage Regeneration. , 2018, Current stem cell research & therapy.

[17]  Ekaterina V. Medvedeva,et al.  Repair of Damaged Articular Cartilage: Current Approaches and Future Directions , 2018, International journal of molecular sciences.

[18]  M. Concha,et al.  Aerogels made of chitosan and chondroitin sulfate at high degree of neutralization: Biological properties toward wound healing. , 2018, Journal of biomedical materials research. Part B, Applied biomaterials.

[19]  M. Doran,et al.  Sheep as a model for evaluating mesenchymal stem/stromal cell (MSC)-based chondral defect repair. , 2018, Osteoarthritis and cartilage.

[20]  A. Oryan,et al.  Effectiveness of chitosan scaffold in skin, bone and cartilage healing. , 2017, International journal of biological macromolecules.

[21]  E. Quenneville,et al.  Effect of a Rapidly Degrading Presolidified 10 kDa Chitosan/Blood Implant and Subchondral Marrow Stimulation Surgical Approach on Cartilage Resurfacing in a Sheep Model , 2017, Cartilage.

[22]  B. Poulet Models to define the stages of articular cartilage degradation in osteoarthritis development , 2017, International journal of experimental pathology.

[23]  R. Karuppal Current concepts in the articular cartilage repair and regeneration. , 2017, Journal of orthopaedics.

[24]  N. Mohan,et al.  Chitosan-hyaluronic acid hydrogel for cartilage repair. , 2017, International journal of biological macromolecules.

[25]  Roberto Raiteri,et al.  Supramolecular Organization of Collagen Fibrils in Healthy and Osteoarthritic Human Knee and Hip Joint Cartilage , 2016, PloS one.

[26]  Ľ. Medvecký,et al.  Effect of enzymatic degradation of chitosan in polyhydroxybutyrate/chitosan/calcium phosphate composites on in vitro osteoblast response , 2016, Journal of Materials Science: Materials in Medicine.

[27]  M. W. Pot,et al.  Improved cartilage regeneration by implantation of acellular biomaterials after bone marrow stimulation: a systematic review and meta-analysis of animal studies , 2016, PeerJ.

[28]  G. Kerkhoffs,et al.  Macroscopic ICRS Poorly Correlates with O Driscoll Histological CartilageRepair Assessment in a Goat Model , 2015 .

[29]  C. Henrist,et al.  Chitosan Enriched Three-Dimensional Matrix Reduces Inflammatory and Catabolic Mediators Production by Human Chondrocytes , 2015, PloS one.

[30]  H. Madry,et al.  Complex and elementary histological scoring systems for articular cartilage repair. , 2015, Histology and histopathology.

[31]  C. Galli,et al.  Chitosan-based scaffold modified with D-(+) raffinose for cartilage repair: an in vivo study , 2015, Journal of Negative Results in BioMedicine.

[32]  H. Madry,et al.  Small Subchondral Drill Holes Improve Marrow Stimulation of Articular Cartilage Defects , 2014, The American journal of sports medicine.

[33]  K. Gupta,et al.  Evaluation of three-dimensional chitosan-agarose-gelatin cryogel scaffold for the repair of subchondral cartilage defects: an in vivo study in a rabbit model. , 2014, Tissue engineering. Part A.

[34]  M. Szychlinska,et al.  New perspectives for articular cartilage repair treatment through tissue engineering: A contemporary review. , 2014, World journal of orthopedics.

[35]  Ľ. Medvecký,et al.  Properties and in vitro characterization of polyhydroxybutyrate–chitosan scaffolds prepared by modified precipitation method , 2014, Journal of Materials Science: Materials in Medicine.

[36]  F. O'Brien,et al.  Advanced Strategies for Articular Cartilage Defect Repair , 2013, Materials.

[37]  Hai-bin Wang,et al.  The support of matrix accumulation and the promotion of sheep articular cartilage defects repair in vivo by chitosan hydrogels. , 2010, Osteoarthritis and cartilage.

[38]  M. Szczodry,et al.  Animal models for cartilage regeneration and repair. , 2010, Tissue engineering. Part B, Reviews.

[39]  Scott A. Rodeo,et al.  The Basic Science of Articular Cartilage , 2009, Sports health.

[40]  R. Boston,et al.  Preclinical animal models in single site cartilage defect testing: a systematic review. , 2008, Osteoarthritis and cartilage.

[41]  M. Shive,et al.  Chitosan-glycerol phosphate/blood implants improve hyaline cartilage repair in ovine microfracture defects. , 2005, The Journal of bone and joint surgery. American volume.

[42]  C. Archer,et al.  Current strategies for articular cartilage repair. , 2005, European cells & materials.

[43]  D. Saris,et al.  The correlation and reproducibility of histological scoring systems in cartilage repair. , 2002, Tissue engineering.

[44]  S. B. Adams,et al.  The surgical anatomy of the stifle joint in sheep. , 1998, Veterinary surgery : VS.

[45]  R. Brentani,et al.  Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections , 1979, The Histochemical Journal.

[46]  O. Benada,et al.  Preclinical alternative model for analysis of porous scaffold biocompatibility in bone tissue engineering. , 2019, ALTEX.

[47]  C. Viegas,et al.  Large Animal Models for Osteochondral Regeneration. , 2018, Advances in experimental medicine and biology.

[48]  Y. Henrotin,et al.  Chitosan: A promising polymer for cartilage repair and viscosupplementation. , 2017, Bio-medical materials and engineering.

[49]  C. McIlwraith,et al.  Joint disease in the horse [2nd Ed.] , 2015 .

[50]  D. Saris,et al.  Evaluation of histological scoring systems for tissue-engineered, repaired and osteoarthritic cartilage. , 2010, Osteoarthritis and cartilage.

[51]  Jerry C. Hu,et al.  The role of tissue engineering in articular cartilage repair and regeneration. , 2009, Critical reviews in biomedical engineering.

[52]  H. Moon,et al.  Chitosan and its derivatives for tissue engineering applications. , 2008, Biotechnology advances.

[53]  D. Lyman Biomedical polymers. , 1968, Annals of the New York Academy of Sciences.

[54]  I. Watt,et al.  JOINT DISEASE , 2022 .