A Polymethyl Methacrylate Method for Large Specimens of Mineralized Bone with Implants

Emmanual, Janson; Hornbeck, Cathy; Bloebaum, Roy D.

doi:10.3109/10520298709108030

Cited by 129 publications

(68 citation statements)

References 8 publications

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“…Mouse IgG (Vector Laboratories) was used as a negative control. Sections through undecalcified tibia were obtained by embedding the whole tibia in methylmethacrylate (Sigma) by using a method previously described (20). Five-micrometer-thick ground cross sections taken through the middle of the subperiosteal bioreactor were stained with von Kossa and Goldner's trichrome and examined under a light microscope.…”

Section: Methodsmentioning

confidence: 99%

In vivo engineering of organs: The bone bioreactor

Stevens

Marini

Schaefer

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

313

208

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Treatment of large defects requires the harvest of fresh living bone from the iliac crest. Harvest of this limited supply of bone is accompanied by extreme pain and morbidity. This has prompted the exploration of other alternatives to generate new bone using traditional principles of tissue engineering, wherein harvested cells are combined with porous scaffolds and stimulated with exogenous mitogens and morphogens in vitro and͞or in vivo. We now show that large volumes of bone can be engineered in a predictable manner, without the need for cell transplantation and growth factor administration. The crux of the approach lies in the deliberate creation and manipulation of an artificial space (bioreactor) between the tibia and the periosteum, a mesenchymal layer rich in pluripotent cells, in such a way that the body's healing mechanism is leveraged in the engineering of neotissue. Using the ''in vivo bioreactor'' in New Zealand White rabbits, we have engineered bone that is biomechanically identical to native bone. The neobone formation followed predominantly an intramembraneous path, with woven bone matrix subsequently maturing into fully mineralized compact bone exhibiting all of the histological markers and mechanical properties of native bone. We harvested the bone after 6 weeks and transplanted it into contralateral tibial defects, resulting in complete integration after 6 weeks with no apparent morbidity at the donor site. Furthermore, in a proof-of-principle study, we have shown that by inhibiting angiogenesis and promoting a more hypoxic environment within the ''in vivo bioreactor space,'' cartilage formation can be exclusively promoted.cartilage ͉ tissue engineering ͉ hard tissue ͉ vascularized organs

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Section: Methodsmentioning

confidence: 99%

In vivo engineering of organs: The bone bioreactor

Stevens

Marini

Schaefer

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

313

208

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“…Specimens were embedded in polymethylmethacrylate, using standard laboratory techniques (Emmanual et al 1987).…”

Section: Implant Processingmentioning

confidence: 99%

Progression of human bone ingrowth into porous-coated implants: Rate of bone ingrowth in humans

Hofmann

Bloebaum

Bachus

1997

Acta Orthopaedica Scandinavica

106

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“…Because embedment using PMMA is the traditional method for preparing mineralized bone tissue and implant for interface analysis after microscopy, the protocol is well established (6)(7)(8). The PMMA-embedrnent procedure enables fixation of mineralized bone specimens with orthopaedic implants in situ.…”

Section: Introductionmentioning

confidence: 99%

A Method for Cemented Bone Interface Examination Without Polymethylmethacrylate Embedment

Bloebaum

Higgins²,

Koller

2006

Journal of Histotechnology

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Although effective, the embedment of bone tissue and orthopaedic devices using polymethylmethacrylate (PMMA) has challenges and limitations. To embed using PMMA, specimens must first be fixed in 70% ethanol, dehydrated in ascending grades of ethanol, and then infiltrated and polymerized in polymethylmethacrylate using standard techniques. This process can take more than 22 d for large bone specimens. Additionally, PMMA embedment has been shown to dissolve bone cement, thus enabling the analysis of the bone-cement interfaces. To conserve processing time while preserving the bone-cement interface, a method was developed for processing mineralized bone tissue in preparation for scanning electron microscopy (SEM) imaging that does not require PMMA embedment. This technique does not require the traditional dehydration and PMMA polymerization process. Instead, fresh mineralized cemented bone specimens were serially sectioned and the marrow removed after formalin fixation. The sections were air-dried then desiccated. The sections were then prepared for SEM imaging and examination. This process takes a fraction of the tissue processing time while not compromising the bone-cement integrity. The SEM image quality was shown to be comparative to images obtained with PMMA-embedded bone specimens.

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A Polymethyl Methacrylate Method for Large Specimens of Mineralized Bone with Implants

Cited by 129 publications

References 8 publications

In vivo engineering of organs: The bone bioreactor

In vivo engineering of organs: The bone bioreactor

Progression of human bone ingrowth into porous-coated implants: Rate of bone ingrowth in humans

A Method for Cemented Bone Interface Examination Without Polymethylmethacrylate Embedment

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