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Engraftment of Prevascularized, Tissue Engineered Constructs in a Novel Rabbit Segmental Bone Defect Model.

Kaempfen A, Todorov A, Güven S, Largo RD, Jaquiéry C, Scherberich A, Martin I, Schaefer DJ - Int J Mol Sci (2015)

Bottom Line: Instead, a variable amount of necrotic tissue formed.Although necrotic area correlated significantly with amount of vessels and the 2-step strategy had significantly more vessels than the 1-step strategy, no significant reduction of necrotic area was found.In conclusion, the animal model developed here represents a highly challenging situation, for which a suitable engineered bone graft with better prevascularization, better resorbability and higher osteogenicity has yet to be developed.

View Article: PubMed Central - PubMed

Affiliation: Clinic for Plastic, Reconstructive, Aesthetic and Hand Surgery, University Hospital Basel, 4031 Basel, Switzerland. alexander.kaempfen@usb.ch.

ABSTRACT
The gold standard treatment of large segmental bone defects is autologous bone transfer, which suffers from low availability and additional morbidity. Tissue engineered bone able to engraft orthotopically and a suitable animal model for pre-clinical testing are direly needed. This study aimed to evaluate engraftment of tissue-engineered bone with different prevascularization strategies in a novel segmental defect model in the rabbit humerus. Decellularized bone matrix (Tutobone) seeded with bone marrow mesenchymal stromal cells was used directly orthotopically or combined with a vessel and inserted immediately (1-step) or only after six weeks of subcutaneous "incubation" (2-step). After 12 weeks, histological and radiological assessment was performed. Variable callus formation was observed. No bone formation or remodeling of the graft through TRAP positive osteoclasts could be detected. Instead, a variable amount of necrotic tissue formed. Although necrotic area correlated significantly with amount of vessels and the 2-step strategy had significantly more vessels than the 1-step strategy, no significant reduction of necrotic area was found. In conclusion, the animal model developed here represents a highly challenging situation, for which a suitable engineered bone graft with better prevascularization, better resorbability and higher osteogenicity has yet to be developed.

No MeSH data available.


Related in: MedlinePlus

(A) Amount of nucleated cells extracted per bone marrow biopsy; each dot represents one donor; (B) Percentage of colony forming units per nucleated cells extracted from bone marrow biopsy; (C) Silicon mold with scaffold during seeding; (D) Tetrazolium (MTT) staining showing the distribution of cells along the periphery after seeding: the top image is a midline section, and the bottom is the outside surface. Black bar represents 1 mm.
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ijms-16-12616-f001: (A) Amount of nucleated cells extracted per bone marrow biopsy; each dot represents one donor; (B) Percentage of colony forming units per nucleated cells extracted from bone marrow biopsy; (C) Silicon mold with scaffold during seeding; (D) Tetrazolium (MTT) staining showing the distribution of cells along the periphery after seeding: the top image is a midline section, and the bottom is the outside surface. Black bar represents 1 mm.

Mentions: Cell isolation yields and percentage of colony forming units from rabbit bone marrow aspirates showed donor variability (Figure 1A,B). The monolayer expansion of isolated primary BMSCs required 25.1 ± 5.4 days to reach approximately two million cells. The seeding efficiency of BMSCs onto the scaffolds using a fibrin hydrogel and a silicon mold (Figure 1C) was always greater than 99%. The cells were found distributed along the periphery of the scaffolds (Figure 1D).


Engraftment of Prevascularized, Tissue Engineered Constructs in a Novel Rabbit Segmental Bone Defect Model.

Kaempfen A, Todorov A, Güven S, Largo RD, Jaquiéry C, Scherberich A, Martin I, Schaefer DJ - Int J Mol Sci (2015)

(A) Amount of nucleated cells extracted per bone marrow biopsy; each dot represents one donor; (B) Percentage of colony forming units per nucleated cells extracted from bone marrow biopsy; (C) Silicon mold with scaffold during seeding; (D) Tetrazolium (MTT) staining showing the distribution of cells along the periphery after seeding: the top image is a midline section, and the bottom is the outside surface. Black bar represents 1 mm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490464&req=5

ijms-16-12616-f001: (A) Amount of nucleated cells extracted per bone marrow biopsy; each dot represents one donor; (B) Percentage of colony forming units per nucleated cells extracted from bone marrow biopsy; (C) Silicon mold with scaffold during seeding; (D) Tetrazolium (MTT) staining showing the distribution of cells along the periphery after seeding: the top image is a midline section, and the bottom is the outside surface. Black bar represents 1 mm.
Mentions: Cell isolation yields and percentage of colony forming units from rabbit bone marrow aspirates showed donor variability (Figure 1A,B). The monolayer expansion of isolated primary BMSCs required 25.1 ± 5.4 days to reach approximately two million cells. The seeding efficiency of BMSCs onto the scaffolds using a fibrin hydrogel and a silicon mold (Figure 1C) was always greater than 99%. The cells were found distributed along the periphery of the scaffolds (Figure 1D).

Bottom Line: Instead, a variable amount of necrotic tissue formed.Although necrotic area correlated significantly with amount of vessels and the 2-step strategy had significantly more vessels than the 1-step strategy, no significant reduction of necrotic area was found.In conclusion, the animal model developed here represents a highly challenging situation, for which a suitable engineered bone graft with better prevascularization, better resorbability and higher osteogenicity has yet to be developed.

View Article: PubMed Central - PubMed

Affiliation: Clinic for Plastic, Reconstructive, Aesthetic and Hand Surgery, University Hospital Basel, 4031 Basel, Switzerland. alexander.kaempfen@usb.ch.

ABSTRACT
The gold standard treatment of large segmental bone defects is autologous bone transfer, which suffers from low availability and additional morbidity. Tissue engineered bone able to engraft orthotopically and a suitable animal model for pre-clinical testing are direly needed. This study aimed to evaluate engraftment of tissue-engineered bone with different prevascularization strategies in a novel segmental defect model in the rabbit humerus. Decellularized bone matrix (Tutobone) seeded with bone marrow mesenchymal stromal cells was used directly orthotopically or combined with a vessel and inserted immediately (1-step) or only after six weeks of subcutaneous "incubation" (2-step). After 12 weeks, histological and radiological assessment was performed. Variable callus formation was observed. No bone formation or remodeling of the graft through TRAP positive osteoclasts could be detected. Instead, a variable amount of necrotic tissue formed. Although necrotic area correlated significantly with amount of vessels and the 2-step strategy had significantly more vessels than the 1-step strategy, no significant reduction of necrotic area was found. In conclusion, the animal model developed here represents a highly challenging situation, for which a suitable engineered bone graft with better prevascularization, better resorbability and higher osteogenicity has yet to be developed.

No MeSH data available.


Related in: MedlinePlus