Limits...
The Axolotl Fibula as a Model for the Induction of Regeneration across Large Segment Defects in Long Bones of the Extremities.

Chen X, Song F, Jhamb D, Li J, Bottino MC, Palakal MJ, Stocum DL - PLoS ONE (2015)

Bottom Line: Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone.By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases.These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, School of Science, Indiana University-Purdue University Indianapolis, Indianapolis, Indiana, United States of America.

ABSTRACT
We tested the ability of the axolotl (Ambystoma mexicanum) fibula to regenerate across segment defects of different size in the absence of intervention or after implant of a unique 8-braid pig small intestine submucosa (SIS) scaffold, with or without incorporated growth factor combinations or tissue protein extract. Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone. By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases. These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

No MeSH data available.


Related in: MedlinePlus

(A) Methylene blue-stained whole mount of a 50% defect three months after treatment with tissue extract.The gap has been completely bridged by an irregular mass of cartilage, which appears to have regenerated primarily from the distal end of the fibula, with only a sliver regenerating from the proximal end (arrow). A cartilage bridge (asterisk), most likely derived from the tibial periosteum, connects the regenerating fibula to the tibia (T). (B) Methylene blue/alizarin red-stained whole mount of a 50% defect three months after treatment with tissue extract. The ends of the fibula were angled with respect to one another so that regeneration from the proximal and distal ends produced a V shape. A supernumerary foot (asterisk) regenerated perpendicular to the fibula. The star indicates the normal foot. Distal is toward the top; proximal is toward the bottom. F = femur; T = tibia; Fb = fibula.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4476796&req=5

pone.0130819.g015: (A) Methylene blue-stained whole mount of a 50% defect three months after treatment with tissue extract.The gap has been completely bridged by an irregular mass of cartilage, which appears to have regenerated primarily from the distal end of the fibula, with only a sliver regenerating from the proximal end (arrow). A cartilage bridge (asterisk), most likely derived from the tibial periosteum, connects the regenerating fibula to the tibia (T). (B) Methylene blue/alizarin red-stained whole mount of a 50% defect three months after treatment with tissue extract. The ends of the fibula were angled with respect to one another so that regeneration from the proximal and distal ends produced a V shape. A supernumerary foot (asterisk) regenerated perpendicular to the fibula. The star indicates the normal foot. Distal is toward the top; proximal is toward the bottom. F = femur; T = tibia; Fb = fibula.

Mentions: Whole limb tissue extract also was effective at inducing regeneration. Of 18 limbs, two had partial regeneration and four had significant regeneration. Figs 13 and 14 show longitudinal sections of limbs in which new cartilage and bone was regenerated over nearly the complete length of the defect. Fig 15 illustrates two other cases treated with tissue extract. In one case, an irregular mass of cartilage has filled the defect and is joined to the tibia by a bridge of cartilage. In the second case, the fibula was regenerated from both distal and proximal stumps. The stumps were angled, so that the regenerated bone formed a shallow V. This specimen had also regenerated a supernumerary foot posteriorly that shared the fibula with the primary foot. The skeletal structures of this specimen are more clearly seen in the micro-CT scan shown in Fig 16.


The Axolotl Fibula as a Model for the Induction of Regeneration across Large Segment Defects in Long Bones of the Extremities.

Chen X, Song F, Jhamb D, Li J, Bottino MC, Palakal MJ, Stocum DL - PLoS ONE (2015)

(A) Methylene blue-stained whole mount of a 50% defect three months after treatment with tissue extract.The gap has been completely bridged by an irregular mass of cartilage, which appears to have regenerated primarily from the distal end of the fibula, with only a sliver regenerating from the proximal end (arrow). A cartilage bridge (asterisk), most likely derived from the tibial periosteum, connects the regenerating fibula to the tibia (T). (B) Methylene blue/alizarin red-stained whole mount of a 50% defect three months after treatment with tissue extract. The ends of the fibula were angled with respect to one another so that regeneration from the proximal and distal ends produced a V shape. A supernumerary foot (asterisk) regenerated perpendicular to the fibula. The star indicates the normal foot. Distal is toward the top; proximal is toward the bottom. F = femur; T = tibia; Fb = fibula.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476796&req=5

pone.0130819.g015: (A) Methylene blue-stained whole mount of a 50% defect three months after treatment with tissue extract.The gap has been completely bridged by an irregular mass of cartilage, which appears to have regenerated primarily from the distal end of the fibula, with only a sliver regenerating from the proximal end (arrow). A cartilage bridge (asterisk), most likely derived from the tibial periosteum, connects the regenerating fibula to the tibia (T). (B) Methylene blue/alizarin red-stained whole mount of a 50% defect three months after treatment with tissue extract. The ends of the fibula were angled with respect to one another so that regeneration from the proximal and distal ends produced a V shape. A supernumerary foot (asterisk) regenerated perpendicular to the fibula. The star indicates the normal foot. Distal is toward the top; proximal is toward the bottom. F = femur; T = tibia; Fb = fibula.
Mentions: Whole limb tissue extract also was effective at inducing regeneration. Of 18 limbs, two had partial regeneration and four had significant regeneration. Figs 13 and 14 show longitudinal sections of limbs in which new cartilage and bone was regenerated over nearly the complete length of the defect. Fig 15 illustrates two other cases treated with tissue extract. In one case, an irregular mass of cartilage has filled the defect and is joined to the tibia by a bridge of cartilage. In the second case, the fibula was regenerated from both distal and proximal stumps. The stumps were angled, so that the regenerated bone formed a shallow V. This specimen had also regenerated a supernumerary foot posteriorly that shared the fibula with the primary foot. The skeletal structures of this specimen are more clearly seen in the micro-CT scan shown in Fig 16.

Bottom Line: Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone.By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases.These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, School of Science, Indiana University-Purdue University Indianapolis, Indianapolis, Indiana, United States of America.

ABSTRACT
We tested the ability of the axolotl (Ambystoma mexicanum) fibula to regenerate across segment defects of different size in the absence of intervention or after implant of a unique 8-braid pig small intestine submucosa (SIS) scaffold, with or without incorporated growth factor combinations or tissue protein extract. Fractures and defects of 10% and 20% of the total limb length regenerated well without any intervention, but 40% and 50% defects failed to regenerate after either simple removal of bone or implanting SIS scaffold alone. By contrast, scaffold soaked in the growth factor combination BMP-4/HGF or in protein extract of intact limb tissue promoted partial or extensive induction of cartilage and bone across 50% segment defects in 30%-33% of cases. These results show that BMP-4/HGF and intact tissue protein extract can promote the events required to induce cartilage and bone formation across a segment defect larger than critical size and that the long bones of axolotl limbs are an inexpensive model to screen soluble factors and natural and synthetic scaffolds for their efficacy in stimulating this process.

No MeSH data available.


Related in: MedlinePlus